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Fiscal Year: FY 2010  Task Last Updated:  02/01/2010 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 03/31/2005  End Date:  03/31/2011 
No. of Post Docs: No. of PhD Degrees: 
No. of PhD Candidates: No. of Master' Degrees:   
No. of Master's Candidates: No. of Bachelor's Degrees: 
No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor: Cucinotta, Francis   Contact Phone:  281-483-0968 
Contact Email: francis.a.cucinotta@nasa.gov 
Flight Program:  
Flight Assignment: NOTE: Received No Cost Extension to 3/31/2011 per PI via JSC (3/2010)

NOTE: Start/end dates changed to 3/31/2005-3/31/2010 per grant documents from PI (12/06)

 

Key Personnel Changes/Previous PI: No Changes to Key Personnel. 
COI Name (Institution): Burma, Sandeep   ( The University of Texas Southwestern Medical Center )
Chen, David   ( The University of Texas Southwestern Medical Center )
Gazdar, Adi   ( The University of Texas Southwestern Medical Center  )
Girard, Luc  ( The University of Texas Southwestern Medical Center  )
Richardson, James   ( The University of Texas Southwestern Medical Center  )
Shay, Jerry   ( The University of Texas Southwestern Medical Center  )
Story, Michael  ( The University of Texas Southwestern Medical Center  )
Wright, Woody  ( The University of Texas Southwestern Medical Center  )
Xie, Xian-Jin  ( The University of Texas Southwestern Medical Center )
Minna, John  ( The University of Texas Southwestern Medical Center )
Yang, Chin-Reng  ( The University of Texas Southwestern Medical Center ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multi-step pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes are being compared in these models to that of gamma-irradiation. These dose risk assessments are designed to measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. They also measure effects on development of cancer “stem-like” cells (“cancer initiating cells”). 3D organotypic culture and animal models also allow measurement and risk estimation in both tissue surrogates and in vivo lung tissues. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORs and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have completed many studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we are completing experiments testing whether transgenic mouse models of lung cancer after low and high LET exposure increase the early onset or progression of lung cancer in the whole animal in real time.

The NSCOR has 4 Projects:

1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation;

2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells;

3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and

4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models.

These projects are supported by 4 Cores: Administrative; Cell Culture; Genomics and Proteomics; and Biostatistics and Bioinformatics. In addition, with funding from the Department of Energy (DOE) contributing to this NSCOR, studies of low-LET and low dose radiation in the same model systems are also being undertaken. The HZE particle irradiation is done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer, a Center for the NCI Early Detection Research Network (EDRN) program, and is part of the NCI sponsored Genetic Epidemiology of Lung Cancer Consortium (GELCC) there is great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits: Lung cancer is the leading cause of cancer death in both women and men in the USA and in the Western world. While lung cancer is caused mostly by smoking exposure, a significant number of lung cancers arise in life time never smokers and thus is a sizable health problem in the USA. Indeed lung cancer in never smokers is the 7th leading cause of world-wide cancer related deaths. Likewise, 50% of all new lung cancer cases occur in former smokers (quitting over 5 years before). In all of these scenarios a major underlying question has been the role of other environmental carcinogens especially environmental radiation. One source of this is radon gas in the environment including radon gas exposure in homes with alpha particles (which represent high-LET radiation). Thus, the interaction of radiation in the environment on Earth including from alpha particles in the genesis of lung cancer and quantitation of the effect of such radiation on lung epithelial cells is of major health importance. In addition, in the past decade in the USA, Europe, and Asia there has been a resurgent interest in radiographic screening for lung cancer, particularly using serial computed tomography (CT) scans. In fact the USA NCI has a large (50,000 person) National Lung Cancer Screening Trial (NLST) that randomized persons to have CT scans or not for the early detection of lung cancer. While there is general agreement that CT scans can detect lung lesions better than conventional chest x-rays, and that CT scans do detect many early stage lung cancers, there are a variety of questions concerning the use of this approach because of its cost and substantial radiation exposure. Greater than 25% of persons screened (current or former smokers) will have some lesions requiring further CT scan follow-up, when biopsies are performed only 1% of the overall group will have histologic evidence of lung cancer. In addition, many of the persons will require several CT scans in follow-up to determine if the lesion is growing to warrant a biopsy. Finally, there have been several analyses that suggest that even in CT detected and resected early stage lung cancers it is not clear that such an approach is actually changing lung cancer mortality in the population screen – the gold standard for proceeding with such an approach as a national health policy. Thus, there is considerable interest in knowing the contribution of CT scan doses of gamma radiation to the pathogenesis of lung cancer. This is particularly true since this radiation is being administered to individuals (current and former smokers) who are at highest risk of having preneoplastic lesions leading to lung cancer. Our studies of low dose radiation of the preclinical human and mouse model systems will allow us to provide some risk assessment for the effect of such radiation doses (low dose) on the genesis of lung cancer and provides the rationale for DOE support of our studies. Radiation is a known carcinogenic influence but the molecular events in lung epithelial cells in response to radiation need to be determined. A key element of this is the impact of radiation on lung epithelial cells with preneoplastic lesions. Our NSCOR uses innovative technology of human bronchial epithelial cells (HBECs) developed by us (coming from 45 different individuals) which have also been engineered to have known oncogenic preneoplastic changes (such as expression of oncogenic K-RASv12, knockdown of expression of mutant p53, expression of oncogenic mutant EGFR, expression of c-myc). Our studies are providing quantitative data following high-LET and low dose gamma irradiation on these HBECs. The primary benefits from this work will be the development of quantitative risk assessment models for astronauts of high-LET radiation in space in order to reduce the uncertainties in prediction of risk of astronauts developing lung cancer as part of long-term missions in space. In addition, as stated we will be developing risk estimates following low dose gamma radiation as seen on Earth, particularly from CT scan studies. Thus, the results will also have major impact and benefit to life on Earth includes new knowledge of the effects on human lung epithelial cells of irradiation in terms of quantitative genetic and epigenetic changes and gene expression changes following irradiation which are of potential importance as markers of radiation exposure that could occur through CT scan screening studies, and environmental exposures, accidentally or through terrorism. A part of this will be to understand the quantitative response of DNA repair pathways to radiation including specific DNA repair components such as DNA repair enzymes and signaling pathways in lung epithelial cells. We are also determining quantitative changes in biologic phenotypes of lung epithelial cells in response to radiation which include colony formation in liquid and soft agar (anchorage independent growth) and differentiation and invasion in 3 dimensional (3D) cultures, and finally growth in vivo as either xenografts (for human cells) or endogenous lung cancers (for mouse models of lung cancer). From all of the above information we will identify novel molecular biomarkers (mRNA, DNA, protein) for lung carcinogenesis. Finally, we are also learning about inter individual variation of the response of HBECs to radiation with respect to the various genetic, epigenetic, mRNA and protein expression, and signaling pathway changes. Our large HBEC panel coupled with new high density single nucleotide polymorphism (SNP) analysis will allow describing polymorphic differences in such responses. Our lung cancer NSCOR is addressing the following specific critical path roadmap and gap questions: "We are improving the understanding of space radiation in lung cancer initiation, promotion and progression spanning cell and molecular biology at the cell, tissue and system biology level that will lead to significant reduction in the uncertainties in risk projection models ; "We are investigating the increased risk from space radiation as a function of age of exposure, age, latency, gender, tissue, radiation quality and dose rate? ; "We are developing tissue specific risk models using human 3D culture and animal models for lung cancer ; "We are investigating how aberrant DNA damage processing, genomic instability, epigenetic effects including methylation, persistent oxidative damage, altered senescence, and non-targeted effects contribute to irradiation-associated lung cancer using a variety of surrogate endpoints that are being validated.

Overall we are conducting research to investigate how biological endpoint and systems biology approaches can be used to integrate research on the cellular and molecular as well as tissue level of radiation damage that will lead to significant reductions in the uncertainties in risk projection models of lung cancer. As will be described in this report, we are confident and will provide compelling evidence that the sum of the efforts of our assembled team are significantly contributing more to assessing cancer risk associated with space radiation compared to the individual components.

 

Task Progress: Project 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation. Michael Story, Ph.D. Lead, Adi F. Gazdar, M.D. Co-Lead, John D. Minna, M.D. Co-Lead.

Specific Aim 1: To quantitate the effects of HZE particle irradiation in causing genetic changes in HBECs and their isogenic derivatives.

Specific Aim 2. To quantitate the amount of epigenetic changes caused by HZE particle irradiation in HBECs and their isogenic derivatives using high throughput quantitative methylation specific PCR (MSP) assays for acquired promoter methylation

Specific Aim 3. To quantitate the effects of HZE particle irradiation in causing expression profile (mRNA and protein) changes in human bronchial epithelial cells (HBECs) and their isogenic derivatives containing various lung cancer-related mutations using array based genome wide and multi protein detecting approaches.

Specific Aim 4. To determine the effect of inter-individual variation on the effects of HZE particle irradiation in causing genetic changes, epigenetic changes, mRNA and protein expression profile changes in human bronchial epithelial cells (HBECs).

Project 1: In the last year this project has focused on 3 specific areas. mRNA Expression Analysis. Comparative gene (mRNA) expression analysis using genome wide array technology at equi-toxic doses of low LET gamma radiation vs. that of HZE radiation (28Si and 56Fe) at different time points. We find that there are different mRNA expression profiles generated by each of the different types of radiation and that this is also dependent on the oncogenotype of the HBECs radiated. DNA Methylation Profiles. While there are no global methylation changes we are exploring genome wide changes using array technology as well as quantitative methylation specific PCR (MSP). miRNA Expression Profiling. We are using genome wide array technology to profile miRNA changes after low and high LET radiation and low dose gamma radiation. Project 1 is now close to hitting the milestones described in the original application. We will very shortly have a set of well characterized genomic responses (both mRNA, DNA copy number, DNA methylation, and miRNA expression) to low LET and HZE radiation (Si, Fe) exposures in a series of isogenic immortalized human bronchial epithelial cells (HBECs) which differ only by specific oncogenic changes introduced (e.g. KRAS, p53, EGFR, E6, E7). These variants thus include cells with altered p53, RAS and EGFR signaling. Our goal is to mine the data available in order to develop biomarkers of response that may be used for relative risk determinations of lung cancer from space radiation exposure. Determining such biomarkers dovetails with the transformation studies of Project 3. With the Biostatistics/Bioinformatics Core (Core D) e are using multiple analysis methodologies to develop biomarkers including Gene Set Enrichment, pathway analysis, smar, and Bayesian analysis for interaction nodes. These biomarkers are also being evaluated in mouse models in Project 4 and can be used to address inter-individual differences in carcinogenic response for risk determination.

Project 2: Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells. David Chen, Ph.D. Project Lead, Sandeep Burma, Ph.D. Co-Lead.

Specific Objectives: Aim 1. To test the hypothesis that the immediate cellular response to DNA damage induced by HZE would be different and more complex than that induced by X-rays.

Aim 2. To test the hypothesis that persistent DNA damage induced by HZE is more complex and difficult to repair than that induced by X-rays and, therefore, more deleterious.

Aim 3. To determine if damage induced by HZE is preferentially repaired by non homologous end joining (NHEJ) or homologous recombination (HR) or is preferentially repaired in certain phases of the cell cycle.

Aim 4. To compare the response of human bronchial epithelial cells (HBECs) as non immortalized, immortalized, and with premalignant oncogenic changes in 3D cultures to HZE exposure using a high throughput approach.

Project 2: This project has had two major developments this year. First development of human lung epithelial cells with endogenous EGFP labeled 53BP1 to allow live cell imaging of DNA repair after low and high LET Radiation in both 2 dimensional (2D) and 3 dimensional (3D). To investigate the cellular responses to DNA double strand breaks (DSBs) induced by low- and high-LET irradiation (IR) in vivo, we generated a human bronchial epithelial cell (HBEC3) line that stably expresses near-physiological levels of EGFP-53BP1 and used this human lung epithelial cell line in a three-dimensional (3D) model tissue culture system (Matrigel) to study induction and repair of DNA DSBs under conditions that were close to those in 3D tissues in vivo by monitoring the formation of 3BP1 foci development and resolution with by monitoring the EGFP labeled endogenous protein. We found that survival curves were identical for HBECs with and without the 53BP1 expression construct. Before we used this system for DSB repair study, we extensively characterized these cells after growing them in Matrigel. These analyses indicated that when we grow HBECs in this extracellular matrix (ECM) culture, they attain a growth-arrested state at day 6, similar to 3D cultures of mammary epithelial cells.

Second, use of HEBECs with endogenous EGFP-53BP1 to study foci dissolution kinetics data after different types of radiation. These studies indicated that a significant proportion of iron-ion-induced damage was irreparable both in 2D and 3D structures, but the number of persistent foci was significantly higher in 3D structures. Collectively, these EGFP-53BP1 foci dissolution analyses suggest that the DNA DSBs induced by high-LET IR are differentially repaired in 2D and 3D structures.

Project 3. Effect of HZE Particle Irradiation on Functional Progression of Human Lung Cancer. John D. Minna, M.D. Project Lead, Jerry Shay, Ph.D. Co-Lead.

Specific Aim 1. Effects of HZE particles on early events in lung cancer progression (soft agar colony formation and other markers of cellular transformation).

Specific Aim 2. Effects of HZE particles on intermediate events in lung cancer progression (invasion in 3 D organotypic cultures).

Specific Aim 3. Effects of HZE particles on late events in lung cancer progression (ability to form tumors in immune deprived mice).

Project 3: We have made substantial progress in the determination of transformation by HZE particles in normal human bronchial epithelial cells (HBEC3KT). (Transformation as defined by the ability to grow in soft agar.) We last reported that there was a peak of transformation (two logs higher from a base rate of 10-7 to ~10-5) for 56Fe particle irradiations at 0.25 Gy which then diminished with higher doses and was more marked than the increase see with gamma radiation at 2-4 Gy. These data are updated in with the addition of 28Si data, which also shows the same response. Interestingly, the HBEC3KT Kras/p53 cell line (which has a four log higher spontaneous transformation rate (10-3) did not show any increase in transformation rate by gamma, Si, or Fe up to 1 Gy. We have now isolated over 160 individual transformed foci and these show that ~50% of the clones breed true for this phenotype. We have studied these and found that they exhibit epithelial to mesenchymal transition (EMT) compared to non irradiated control transformed foci by both morphology and molecular markers (vimentin, ecadherin expression). In addition, they have an altered radiation survival curve phenotype. With these cell isolates in hand we will begin to determine tumorigenicity. Tumors themselves will be examined for radio sensitivity and aggressiveness. Tumor phenotype will be compared to those mouse lung carcinomas generated after Fe-exposure in Project 4, which were characterized as especially aggressive. Those cells that develop tumors will be characterized at the genomic and proteomic level and compared against the phenotype of human lung tumors generated via smoking or tumors seen in non-smokers, or via radiation-exposure, in particular via inhalation of radioisotopes such as Pu or Rn. This should allow the characterization and risk for radiation-induced lung cancers, particularly those induced by HZE exposure.

Project 4. Effects of HZE Particles on the Development of Lung Cancer in Novel Mouse Models. Jerry W. Shay, Ph.D. Project Lead, James Richardson, DVM, Ph.D. Co-Investigator, Oliver Delgado, Student Assistant.

Specific Aim 1. Use the K-RAS transgenic mouse model of human lung cancer to determine effects of low- and high-LET radiation in tumor initiation and progression.

Specific Aim 2. Use additional transgenic mouse models to determine the effects of low- and high-LET radiation in both living animals and in isolated tissues.

Summary of Project 4: 1. Fractionated and single dose irradiation of 1Gy 56Fe- particles do not increase tumor incidence

2. Tumors in the LA1 K-RAS mice irradiated with fractionated doses of (0.2Gy/day, total 1 Gy (1GeV/n) 56Fe- particles have a statistically significant increased probability of progressing to invasive carcinoma in comparison to a single acute 1Gy 56Fe- dose.

3. These results suggest significant effects of fractionated high-LET irradiation on cancer progression

4. To test this hypothesis we have initiated additional single dose and fractionated doses to confirm these important observations. We have irradiated mice with fractionated x-ray (0.6 Gy/day x 5 days) and have not observed an increase incidence of invasive tumors. We have initiated fractionated 56Fe- particle experiments (0.1 Gy/day x 10 over a 12 day period).

5. We are testing the hypothesis that fractionated high-LET irradiation induces persistent inflammatory signaling that does not occur in single doses. An alternative hypothesis is that there could be a role of stem cell repopulation for lung tissue that is important in cancer progression. Thus, acute doses of high-LET ions may increase cell killing, reducing the number of target stem cells available for initiation and progression to lung cancer.

6. We have provided non lung tissues from our control and irradiated mice to other investigators (Ameila Eisch, brain tissue; Zhi-Min Yuan, kidney, liver, spleen)

7. Mouse telomerase knockout model of lung cancer. Our rationale and overall goal is to produce a mouse with “humanized” telomere length so that we can irradiate with X-ray, proton and HZE particles to determine genomic changes and DNA damage/repair in the lung and other tissues. Inbred strains of mice have extraordinarily long telomeres and thus much of the genomic instability due to telomeres shortening in humans is unlikely to be observed in typical mouse irradiation studies. We have reached the third generation of breeding mTERT KO to KO to obtain short (more human like) telomere length and initiated our first experiments in the Fall 2009 NSRL run. These animals have been returned from the BNL and we will be conducting pilot microarray experiments to compare to similarly irradiated wild-type mice for DNA damage and inflammatory changes that may distinguish mice with humanized telomeres from typical inbred strains of mice. Future experiments will be initiated to determine if more end joining, end-end fusions, cytogenetic alterations, micronuclei, ongoing reactive oxygen species (ROS) occur when these mice are exposed to low- and high-LET irradiation.

8. EGFR tyrosine kinase domain mutation mouse/ Our rationale for initiating experiment with a new mouse model is that astronauts do not smoke. EGFR mutations are frequently found in lung cancers from never smokers. The models we have chosen to use have an EGFR mutation or small deletion that can be induced by adding doxycycline in the food or drinking water. We now have the mice in our breeding colony and have been scaling up breeding pairs. Experiments under consideration when sufficient numbers of animals are available include the following: We will activate the EGFR-mut about a month prior to low- and high-LET irradiation to compare DNA damage and repair in tumor versus normal tissue; We will also subject these animals to low- and high-LET irradiation prior to activating the EGFR mutation and then add doxycycline to the food for three months. These experiments are designed to further dissect stromal/epithelial differences in cancer progression. The hypothesis that we propose to test is that by activating the stromal compartment prior to tumor initiation the host environment may be permissive for the future growth of the tumors; We will compare the EGFR-mut inducible model to the inducible K-RAS mouse model.

9. DOE Related studies: One aspects of our NSCOR is supported by the DOE. While low doses or even low dose rates have not yielded informative animals results in most instances, we have now formulated a series of testable hypothesis that may be tenable. We propose to send both the mutant EGFR and K-RAS mice to the Colorado State University (CSU) low dose facility and then expose them to low dose rates (10 cGy/day) for 10, 20 and 30 days then activate mutant EGFR or K-RAS. We will be able to address the central and unanswered question if tumors appear faster upon activation of the oncogene after low dose exposure to the stroma). We can also determine if activated tumor will or will not regress when the doxycycline food is removed at 90 days.

Core A: Administrative Core. Leads: John Minna and Jerry Shay, and UTSW NSCOR Administrator: Brenda Zielke.

The Administrative Core has established accounts and control over disbursement of all funds from this NSCOR application. It has also kept track of all personnel employed by this NSCOR. It has provided a central place for communication and coordinated data sharing In addition, it has established a series of biweekly to monthly scientific and planning meeting of the NSCOR investigators at UTSW. Approximately 25 scientific personnel attend each of these meetings. At each meeting administrative matters are discussed, experiments are planned, administrative changes made by the senior leadership team, and most importantly a scientific presentation by one of the NSCOR investigators is made and critiqued by the other investigators. Dr. Minna serves as the Administrative Core Lead with Dr. Jerry Shay as the Administrative Core Co-Lead. The Administrative Core coordinated all of the arrangements for the participation of several NSCOR investigators for the NASA reverse site visit February 18-19, 2009, and travel for several investigators to the 20th NASA investigators meeting in Cologne Germany. The Administrative core also arranged the Internal Scientific Advisory Board Review (March 4, 2008 as part of a mini-retreat at UT Southwestern) and the External Advisory Board Review (October 15, 2008 Mary Helen Barcellos-Hoff, Robert Ullrich and Martin Brown participating). Finally, the Administrative Core coordinated travel of multiple personnel to BNL for experimental work.

Core B. Cell Culture. Core Leads: John D. Minna, M.D. and Jerry Shay, Ph.D.

Enhancement of human bronchial epithelial cell 3D lung model of tissue like development. There is increasing evidence that cancer arises from stem cell populations that are important for the normal turnover of tissues and the rapid repair of tissues in instances of injury. In the lung there is evidence for a bronchiole alveolar stem cell (BASC) population that may give rise to lung carcinoma. We developed a panel of immortalized human bronchial epithelial cells and have previously demonstrated that these cells when placed in an air medium interface overlying a fibroblast stromal collagen matrix could differentiate into mucous and ciliated bronchial epithelial cells. These results were expected since the cells originated from the central airways and were obtained from mid size bronchial tissues. We examined these cells for a variety of differentiated cell markers and observed that they expressed p63 (a transit amplifying cell marker) and surprisingly surfactant protein A (SP-A), CC10, a Clara cell marker, and when differentiated weak expression of the thyroid transcription factor TITF1 (known to be important in adenocarcinomas of the peripheral airway of the lungs). These peripheral markers associated with Clara and type II alveolar cells imply that our immortalized HBECs have enhanced plasticity and should be considered pluripotent lung stem cells. We next placed these cells into Matrigel overlying a lung fibroblast (IMR90) feeder layer and observed that the cells could efficiently differentiate into spheroids or cyst like structures. Studies of these HBECs growing in Matrigel and forming cysts and SACs indicated they started to express SP-A and morphologically resembled small bronchioles and alveolar sacs. When examined in the electron microscope, these cells also had lamellar bodies which are a hallmark of type II alveolar cells. This new improved 3D model of lung cell differentiation has many advantages over the previous model. First, they can differentiate in 3-5 days in vitro making it feasible to set these up at BNL for future NSRL runs. This is in contrast to the air liquid interface for differentiation into ciliated and mucous secreting bronchial cells which require a minimum of two weeks of culture and our previous experience indicated that they do not ship well to BNL. Second, these differentiating small airways cells resemble the type of lung cancer that is likely to be increased in astronauts on long-term space missions. Thus, during the 2008 and 2009 NSRL runs we initiated our irradiation experiments with these cells (see Project 2 overview). Importantly, we introduced a reporter gene 53BP1 containing EGFP so that they sites of irradiation induced damage could be visualized in real time. One final improvement in the 3D culture of our immortalized HBECs consists of culturing them on top of Matrigel. Under these conditions, the cells form web like structures then bud as if they are formed alveoli. This new method also permits a way to have 3D cultures in 5 days and thus promises to be a new addition to our arsenal of ways to examine the influence of irradiation on lung cells under various conditions.

Core C. Genomics and Proteomics Core. Core Lead: Michael Story, Ph.D., Ching-Reng Yang, Ph.D. Core Co-Lead.

Genomics Core: The Core has continued to perform genome wide profiling including mRNA, miRNA, DNA SNP copy number, and DNA methylation, as well as reverse phase protein array (RPPA) profiling on samples before and after radiation in support of all of the projects of this NSCOR. In addition, this Core interfaces with the Biostatistics and Bioinformatics/Database Core (Core D) to ensure the primary results from these genome wide studies are available for appropriate biostatistical and bioinformatics analysis. These are very precise technologies that requires extensive experience to perform and sophisticated instrumentation to carry out. Furthermore, having a single facility through which all array analysis passes minimizes the variability brought about by different individuals processing samples even though the same platform is being used. For these reasons we included a Core facility (Core C, Genomics and Proteomics) in this NSCOR grant. This NSCOR Core also interfaces with the UTSW Cancer Center Microarray and the UTSW school wide Cores.

The NSCOR Core has the following duties: 1. Interact with Project Leaders and investigators in the design of experiments that use microarray technologies (mRNA, miRNA, DNA copy number, and proteomics) in their experimental approach. Biostatistics and bioinformatics will also be integrated in this process. 2. Establish procedures for consistency in sample preparation and RNA, DNA, and protein extraction, test all RNA, DNA, and protein samples for integrity, and provide reports to that effect. 3. Perform all hybridizations, perform all scans, and place data on a secure server. 4. Provide early analysis on array quality control and preliminary data analysis to investigators. 5. Interact with the Biostatistics and Bioinformatics Core (Core D) to provide bioinformatic analysis of data according to project requirements.

NSCOR Core D: Biostatistics and Bioinformatics. Core Lead: Xian-Jin Xie, Ph.D.; Core Co-Lead: Luc Girard, Ph.D. and Yang Xie, Ph.D.

The Biostatistics and Bioinformatics Core (Core D) provides comprehensive biostatistics and bioinformatics expertise to ensure the statistical integrity and to optimize data analysis of the University of Texas Southwestern (UTSW) NSCOR studies. In doing so, Core D assists investigators with testable hypothesis formulations and proper planning of experiments, manages experimental data generated by NSCOR projects, conducts and advises data analysis, and assists in interpreting final results. To play an integrated role and to serve all the NSCOR projects in a timely manner, Core D biostatisticians Drs. Xian-Jin Xie, Luc Girard, Yang Xie, Chul Ahn and Song Zhang have participated in all monthly project meetings, ensuring that proper biostatistics and data management considerations are adequately taken during all phases of planned experiments. In conjunction with the Genomics (Expression Profiling and Proteomics) Core (Core C) and the Cell Culture Core (Core B), the Biostatistics and Bioinformatics Core (Core D) has the following specific aims:

- To provide database support and expertise for the collection, storage, integration, and retrieval of all sorts of data generated from NSCOR projects.

- To provide valid and optimal statistical design and to conduct proper analysis required to address the specific aims of each project.

- To review background and rationale, to assist in the design, evaluation, and analysis of new research proposal arising from the individual projects.

- To assist the other Cores and Projects in the proper analysis and interpretations of different types of bioassays, particularly mega-data such as those generated from gene expression arrays.

- To assist in manuscript preparation and to review scientific submissions.

- To assist the NSCOR team in building radiation cancer risk models.

As we progress on all the NSCOR projects, significant amount of new data have been generated since our 2008 annual progress report. As a result of Core D effort, a comprehensive data management system has been established that allows efficient access and sharing of all NSCOR data. Data Collection Close to 10,000 samples (including cell pellets and cryogenically preserved cell lines) and experiments (irradiation, microarrays, mouse data) have so far been collected and stored in the NSCOR database. These data can be viewed or edited by any participating investigator provided they have the necessary permissions.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Asaithamby A, Chen DJ. "Spatial and temporal characterization of DNA damage response to high and low-LET radiation in living cells." Presented at the NASA human research program investigators’ workshop, League City, Texas, February 2-4, 2009.

NASA human research program investigators’ workshop, February 2009. , Feb-2009

Abstracts for Journals and Proceedings Asaithamby A, Chen DJ. "Cellular responses to DNA double-strand breaks after low-dose gamm-irradiation." Presented at the Low-dose radiation research investigators' workshop VIII, Bethesda, Maryland, April 2009.

Low-dose radiation research investigators' workshop VIII, Bethesda, Maryland, April 2009. , Apr-2009

Abstracts for Journals and Proceedings Burong Hu, Asaithamby A, Chen DJ. "Cellular responses to clustered DNA damages induced by high energy and high-Z particles irradiation." Presented at the 55th Annual Radiation Research Society meeting, Savannah, Georgia, October 4-7, 2009.

55th Annual Radiation Research Society meeting, Savannah, Georgia, October 4-7, 2009. , Oct-2009

Abstracts for Journals and Proceedings Asaithamby A, Chen DJ. "Live cell imaging approach to directly monitor induction and repair of DNA damages generated by low- and high-linear energy transfer irradiation." Invited talk, 55th Annual Radiation Research Society meeting, Savannah, Georgia, October 4-7, 2009.

55th Annual Radiation Research Society meeting, Savannah, Georgia, October 4-7, 2009. , Oct-2009

Abstracts for Journals and Proceedings Asaithamby A, Burong Hu, Chen DJ. "Cellular responses to DNA damages induced by low- and high-linear energy transfer irradiation in monolayer and three dimensional cell cultures." Invited Talk presented at “Radiation Biology and Radiation Protection”, Fudan University, Shanghai, China, October 14-16, 2009.

Radiation Biology and Radiation Protection, Fudan University, Shanghai, China, October 14-16, 2009. , Oct-2009

Abstracts for Journals and Proceedings Rodriguez N, Krysan K, Shay JW, Minna JD, Dubinett SM. "Hepatocyte Growth Factor alters the expression the Sim1 transcription factor in NSCLC." IASLC conference (International Association for the Study of Lung Cancer), San Francisco, CA, July 31-August 4, 2009.

IASLC conference (International Association for the Study of Lung Cancer), San Francisco, CA, July 31-August 4, 2009. , Aug-2009

Abstracts for Journals and Proceedings Christodoulidou A, Chiourea M, Raftopoulou C, Papaioannou CK, Hoshiyama H, Wright WE, Shay JW, Gagos S. "Opposing roles of telomerase in the generation of polyploidy during continuous neoplastic cell growth." Cold Spring Harbor Laboratory annual meeting, Cold Spring Harbor, NY, May 2009.

Cold Spring Harbor Laboratory annual meeting, Cold Spring Harbor, NY, May 2009. , May-2009

Abstracts for Journals and Proceedings Delgado O, Richardson JA, Xie X-J, Wright WE, Story MD, Minna JD, Shay JW. "Invasive Cancer Induced by Radiation in the LA1 K-ras Mouse Model of Lung Cancer." Heavy Ions in Therapy and Space Symposium 2009, Cologne, Germany, July 6-10, 2009.

Heavy Ions in Therapy and Space Symposium 2009, Cologne, Germany, July 6-10, 2009. , Jul-2009

Abstracts for Journals and Proceedings Hazra S, Gardner BK, Krysan K, Walser TC, Brothers J 2nd, Larsen JE, Pertsemlidis A, Shay JW, Minna JD, Spira A, Dubinett SM. "The role of micro RNA 125a-3P in the pathogenesis of lung cancer." IASLC conference (International Association for the Study of Lung Cancer), San Francisco, CA, July 31-August 4, 2009.

IASLC conference (International Association for the Study of Lung Cancer), San Francisco, CA, July 31-August 4, 2009. , Aug-2009

Articles in Peer-reviewed Journals Zhao Y, Sfeir AJ, Zou Y, Buseman CM, Chow TT, Shay JW, Wright WE. "Telomere extension occurs at most chromosome ends and is uncoupled from fill-in in human cancer cells." Cell. 2009 Aug 7;138(3):463-75. PubMed PMID: 19665970 , Aug-2009
Articles in Peer-reviewed Journals Vaughan MB, Ramirez RD, Andrews CM, Wright WE, Shay JW. "H-ras expression in immortalized keratinocytes produces an invasive epithelium in cultured skin equivalents." PLoS One. 2009 Nov 19;4(11):e7908. PubMed PMID: 19936293 , Nov-2009
Articles in Peer-reviewed Journals Yoon H, Kim DJ, Ahn EH, Gellert GC, Shay JW, Ahn CH, Lee YB. "Antitumor activity of a novel antisense oligonucleotide against Akt1." J Cell Biochem. 2009 Nov 1;108(4):832-8. PubMed PMID: 19693774 , Nov-2009
Articles in Peer-reviewed Journals Sullivan JP, Minna JD, Shay JW. "Evidence for self-renewing lung cancer stem cells and their implications in tumor initiation, progression, and targeted therapy." Cancer Metastasis Rev. 2010 Jan 23. [Epub ahead of print] PubMed PMID: 20094757 , Jan-2010
Articles in Peer-reviewed Journals Asaithamby A, Chen DJ. "Cellular responses to DNA double-strand breaks after low-dose gamma-irradiation." Nucleic Acids Res. 2009 Jul;37(12):3912-23. PubMed PMID: 19401436 , Jul-2009
Articles in Peer-reviewed Journals Asaithamby A, Uematsu N, Chatterjee A, Story MD, Burma S, Chen DJ. "HZE particle-induced DNA double-strand break repair in normal human fibroblasts." Radiation Research (in press), January 2010. , Jan-2010
Articles in Peer-reviewed Journals Xie Y, Wang X, Story M. "Statistical methods of background correction for Illumina BeadArray data." Bioinformatics. 2009 Mar 15;25(6):751-7. Epub 2009 Feb 4. PubMed PMID: 19193732 , Mar-2009
Articles in Peer-reviewed Journals Roig AI, Hight SK, Minna JD, Shay JW, Rusek A, Story MD. "DNA damage intensity in fibroblasts in a 3-dimensional collagen matrix correlates with the Bragg curve energy distribution of a high LET particle." Int J Radiat Biol. 2010 Mar;86(3):194-204. PubMed PMID: 20201648 , Mar-2010
Articles in Peer-reviewed Journals Allen JD, Chen M, Xie Y. "Model-based bkground correction (MBCB): R methods and GUI for Illumina Bead-Array data." Journal of Cancer Science & Therapy 2009 Nov;1(1): 025-027. http://dx.doi.org/10.4172/1948-5956.1000004 , Nov-2009
Articles in Peer-reviewed Journals Hu F, Schucany W, Ahn C. "Nonparametric sample size estimation for diagnostic tests with multiple observations per subject." Drug Information Journal, in press, January 2010. , Jan-2010
Awards Burong Hu. "Scholar in training award to attend 55th annual Radiation Research Society annual meeting, October 2009." Oct-2009
Awards Delgado O. "Third place oral presentation at Heavy Ions in Therapy and Space Symposium 2009, Cologne, Germany, July 6-10, 2009." Jul-2009
Awards Camacho C. "International Workshop on Space Radiation Research, Cologne, Germany, July 2009." Jul-2009
Dissertations and Theses Delgado O. "Effects of radiation exposure on lung carcinogenesis." PhD defense, The University of Texas Southwestern Medical Center, degree awarded December 2, 2009. , Dec-2009
Significant Media Coverage Organized by UT Southwestern NSCOR. "Space Travel Radiation and Health - Community Outreach Educational Program." Attending ~250 including: Honorable US Congressman Ralph M. Hall, Richard Scheuring, Medical Operations NASA-JSC, Francis Cucinotta, Chief Scientist Space Radiation Program, NASA-JSC, and scientists from UT Southwestern and over 100 high school students, October 2009., Oct-2009
 
Fiscal Year: FY 2009  Task Last Updated:  02/04/2009 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 03/31/2005  End Date:  03/31/2010 
No. of Post Docs: No. of PhD Degrees: 
No. of PhD Candidates: No. of Master' Degrees:   
No. of Master's Candidates: No. of Bachelor's Degrees: 
No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor: Cucinotta, Francis   Contact Phone:  281-483-0968 
Contact Email: francis.a.cucinotta@nasa.gov 
Flight Program:  
Flight Assignment: NOTE: Start/end dates changed to 3/31/2005-3/31/2010 per grant documents from PI (12/06)

 

Key Personnel Changes/Previous PI: No Changes to Key Personnel. 
COI Name (Institution): Burma, Sandeep   ( The University of Texas Southwestern Medical Center )
Chen, David   ( The University of Texas Southwestern Medical Center )
Gazdar, Adi   ( The University of Texas Southwestern Medical Center  )
Girard, Luc  ( The University of Texas Southwestern Medical Center  )
Richardson, James   ( The University of Texas Southwestern Medical Center  )
Shay, Jerry   ( The University of Texas Southwestern Medical Center  )
Story, Michael  ( The University of Texas Southwestern Medical Center  )
Wright, Woody  ( The University of Texas Southwestern Medical Center  )
Xie, Xian-Jin  ( The University of Texas Southwestern Medical Center )
Minna, John  ( The University of Texas Southwestern Medical Center )
Yang, Chin-Reng  ( The University of Texas Southwestern Medical Center ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multi-step pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes are being compared in these models to that of gamma-irradiation. These dose risk assessments are designed to measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. They also measure effects on development of cancer “stem-like” cells (“cancer initiating cells”). 3D organotypic culture and animal models also allow measurement and risk estimation in both tissue surrogates and in vivo lung tissues. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORs and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have completed many studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we are completing experiments testing whether transgenic mouse models of lung cancer after low and high LET exposure increase the early onset or progression of lung cancer in the whole animal in real time. The NSCOR has 4 Projects: 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation; 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells; 3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and 4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models. These projects are supported by 4 Cores: Administrative; Cell Culture; Genomics and Proteomics; and Biostatistics and Bioinformatics. In addition, with funding from the Department of Energy (DOE) contributing to this NSCOR, studies of low-LET and low dose radiation in the same model systems are also being undertaken. The HZE particle irradiation is done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer, a Center for the NCI Early Detection Research Network (EDRN) program, and is part of the NCI sponsored Genetic Epidemiology of Lung Cancer Consortium (GELCC) there is great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits: Lung cancer is the leading cause of cancer death in both women and men in the USA and in the Western world. While lung cancer is caused by mostly by smoking exposure, a significant number of lung cancers arise in life time never smokers and thus is a sizable health problem in the USA. Indeed lung cancer in never smokers is the 7th leading cause of world-wide cancer related deaths. Likewise, 50% of all new lung cancer cases occur in former smokers (quitting over 5 years before). In all of these scenarios a major underlying question has been the role of other environmental carcinogens especially environmental radiation. One source of this is radon gas in the environment including radon gas exposure in homes with alpha particles (which represent high-LET radiation). Thus, the interaction of radiation in the environment on Earth including from alpha particles in the genesis of lung cancer and quantitation of the effect of such radiation on lung epithelial cells is of major health importance. Over the past 10 years there has been a resurgence of interest in the use of computed tomography (CT) scanning for the early detection of lung cancer. Currently we are awaiting the results of the National Cancer Institute (NCI) National Lung Cancer Screening Trial (NLST) which randomized ~55,000 current and former smokers to be screened regularly with CT scans on a yearly basis or not to determine if such screening would impact lung cancer mortality. While there is a lot of debate about this area, this trial should resolve this important issue. If the decision is to use CT scans for screening in the general population there will be a large increase in the exposure of potentially individuals at very high risk of developing lung cancer (current and former smokers) to repeated doses of radiation from the CT scans. This falls into to the area of low dose radiation exposure covered by the Department of Energy (DOE) supported part of this application. Thus, our proposed low dose and low dose rate studies of gamma radiation in the HBEC and mouse models of lung cancer Projects are of great immediate relevance to developing risk models for such radiation exposure in the USA. Radiation is a known carcinogenic influence but the molecular events in lung epithelial cells in response to radiation need to be determined. A key element of this is the impact of radiation on lung epithelial cells with preneoplastic lesions. Our NSCOR uses innovative technology of human bronchial epithelial cells (HBECs) developed by us (coming from 45 different individuals) which have also been engineered to have known oncogenic preneoplastic changes (such as expression of oncogenic K-RASv12, knockdown of expression of mutant p53, expression of oncogenic mutant EGFR, expression of c-myc).

Our studies are providing quantitative data following high-LET irradiation on these HBECs. The primary benefits from this work will be the development of quantitative risk assessment models for astronauts of high-LET radiation in space in order to reduce the uncertainties in prediction of risk of astronauts developing lung cancer as part of long-term missions in space However, the results will also have major impact and benefit to life on Earth includes new knowledge of the effects on human lung epithelial cells of irradiation in terms of quantitative genetic and epigenetic changes and gene expression changes following irradiation which are of potential importance as markers of radiation exposure that could occur through environmental exposures, accidentally or through terrorism. As part of this will be the quantitative response of DNA repair pathways to radiation including specific DNA repair components such as DNA repair enzymes and signaling pathways in lung epithelial cells. We are also determining quantitative changes in biologic phenotypes of lung epithelial cells in response to radiation which include colony formation in liquid and soft agar (anchorage independent growth) and differentiation and invasion in 3 dimensional cultures, and finally growth in vivo as either xenografts (for human cells) or endogenous lung cancers (for mouse models of lung cancer). From all of the above information we will identify novel molecular biomarkers (mRNA, DNA, protein) for lung carcinogenesis. Finally, we are also learning about inter individual variation of the response of HBECs to radiation with respect to the various genetic, epigenetic, mRNA and protein expression, and signaling pathway changes. Our large HBEC panel coupled with new high density single nucleotide polymorphism (SNP) analysis will allow describing polymorphic differences in such responses.

Our lung cancer NSCOR is addressing the following specific critical path roadmap and gap questions:

•We are improving the understanding of space radiation in lung cancer initiation, promotion and progression spanning cell and molecular biology at the cell, tissue and system biology level that will lead to significant reduction in the uncertainties in risk projection models

•We are investigating the increased risk from space radiation as a function of age of exposure, age, latency, gender, tissue, radiation quality and dose rate?

•We are developing tissue specific risk models using human 3D culture and animal models for lung cancer

•We are investigating how aberrant DNA damage processing, genomic instability, epigenetic effects including methylation, persistent oxidative damage, altered senescence, and non-targeted effects contribute to irradiation-associated lung cancer using a variety of surrogate endpoints that are being validated.

Overall we are conducting research to investigate how biological endpoint and systems biology approaches can be used to integrate research on the cellular and molecular as well as tissue level of radiation damage that will lead to significant reductions in the uncertainties in risk projection models of lung cancer. As will be described in this report, we are confident and will provide compelling evidence that the sum of the efforts of our assembled team are significantly contributing more to assessing cancer risk associated with space radiation compared to the individual components.

 

Task Progress: Project 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation. Michael Story, Ph.D. Lead, Adi F. Gazdar, M.D. Co-Lead, John D. Minna, M.D. Co-Lead.

Specific Aim 1: To quantitate the effects of HZE particle irradiation in causing genetic changes in HBECs and their isogenic derivates.

Specific Aim 2. To quantitate the amount of epigenetic changes caused by HZE particle irradiation in HBECs and their isogenic derivates using high throughput quantitative methylation specific PCR (MSP) assays for acquired promoter methylation

Specific Aim 3. To quantitate the effects of HZE particle irradiation in causing expression profile (mRNA and protein) changes in human bronchial epithelial cells (HBECs) and their isogenic derivates containing various lung cancer-related mutations using array based genome wide and multi protein detecting approaches.

Specific Aim 4. To determine the effect of inter-individual variation on the effects of HZE particle irradiation in causing genetic changes, epigenetic changes, mRNA and protein expression profile changes in human bronchial epithelial cells (HBECs).

Project 1: In the last year this project has focused on 3 specific areas. mRNA Expression Analysis. The first area is comparative gene (mRNA) expression analysis at equi-toxic doses of low LET radiation vs that of HZE radiations, specifically 56Fe. DNA Methylation Profiles. The second area of emphasis this last year was the analysis of radiation-induced changes in DNA methylation patterns. miRNA Expression Profiling. The third area of emphasis dovetails with our gene expression analysis, and represents a relatively new area of research for radiation exposures.

Project 2: Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells. David Chen, Ph.D. Project Lead, Sandeep Burma, Ph.D. Co-Lead.

Specific Objectives :

Aim 1. To test the hypothesis that the immediate cellular response to DNA damage induced by HZE would be different and more complex than that induced by X-rays.

Aim 2. To test the hypothesis that persistent DNA damage induced by HZE is more complex and difficult to repair than that induced by X-rays and, therefore, more deleterious.

Aim 3. To determine if damage induced by HZE is preferentially repaired by non homologous end joining (NHEJ) or homologous recombination (HR) or is preferentially repaired in certain phases of the cell cycle.

Aim 4. To compare the response of human bronchial epithelial cells (HBECs) as non immortalized, immortalized, and with premalignant oncogenic changes in 3D cultures to HZE exposure using a high throughput approach.

Summary of Current Progress/Detailed Results

1. Damage Responses of HBECs Irradiated with Fe Particles.

2. Repair kinetics of DNA damage induced by Fe particles in HBECs

3. DNA damage responses in organotypic 3D cultures of HBECs.

4. Establishment of a YFP-53BP1 system for live cell imaging of DNA damage and its repair:

5. Recruitment of YFP-53BP1 to the sites of DNA DSB in living cells is dose-dependent:

6. Live cell imaging of repair of DNA DSBs induced by gamma-radiation:

7. Expression of EGFP-53BP1 in HBEC3KT cells:

8. EGFP-53BP1 forms foci in 3D cultures of HBEC3KT cells.

9. To compare the ability of monolayer and 3D cultures of HBECs to carry out repair of DSBs induced by gamma rays or HZE by live cell imaging.

Project 3. Effect of HZE Particle Irradiation on Functional Progression of Human Lung Cancer. John D. Minna, M.D. Project Lead, Jerry Shay, Ph.D. Co-Lead.

Specific Aim 1. Effects of HZE particles on early events in lung cancer progression (soft agar colony formation and other markers of cellular transformation).

Specific Aim 2. Effects of HZE particles on intermediate events in lung cancer progression (invasion in 3 D organotypic cultures).

Specific Aim 3. Effects of HZE particles on late events in lung cancer progression (ability to form tumors in immune deprived mice).

Immediate and long-term plans:

Project 1 is now very close to hitting the milestones described in the original application. We will very shortly have a set of well characterized genomic responses (both gene and miRNA expression) to low LET and HZE radiation exposures in a series of isogenic immortalized human bronchial epithelial cells (HBECs) which differ only by specific oncogenic changes introduced (e.g. KRAS, p53, EGFR, E6, E7). These variants thus include cells with altered p53, RAS and EGFR signaling. Our goal will be to mine the data available in order to develop biomarkers of response that may be used for relative risk determinations of lung cancer from space radiation exposure. Determining such biomarkers dovetails with the transformation studies of Project 3. We will use multiple analysis methodologies to develop biomarkers including Gene Set Enrichment, pathway analysis, smar, and in particular, Bayesian analysis for interaction nodes. In future studies these biomarkers can be evaluated in mouse models and in particular, can be used to address inter-individual differences in carcinogenic response for risk determination.

Project 4. Effects of HZE Particles on the Development of Lung Cancer in Novel Mouse Models. Jerry W. Shay, Ph.D. Project Lead, James Richardson, DVM, Ph.D. Co-Investigator, Oliver Delgado, Student Assistant.

Specific Aim 1. Use the K-RAS transgenic mouse model of human lung cancer to determine effects of low- and high-LET radiation in tumor initiation and progression.

Specific Aim 2. Use additional transgenic mouse models to determine the effects of low- and high-LET radiation in both living animals and in isolated tissues.

Specific Aim 1. Use the K-RAS transgenic mouse model of human lung cancer to determine effects of low- and high-LET radiation in tumor initiation and progression.

Summary:

1. Fractionated and single dose irradiation of 1Gy 56Fe- particles do not increase tumor incidence

2. Tumors in the LA1 K-RAS mice irradiated with fractionated doses of (0.2Gy/day, total 1 Gy (1GeV/n) 56Fe- particles have a statistically significant increased probability of progressing to invasive carcinoma in comparison to a single acute 1Gy 56Fe- dose.

3. These results suggest significant effects of fractionated high-LET irradiation on cancer progression

4. To test this hypothesis we have initiated additional single dose and fractionated doses to confirm these important observations. We have irradiated mice with fractionated x-ray (0.6 Gy/day x 5 days) and have not observed an increase incidence of invasive tumors. We have initiated fractionated 56Fe- particle experiments (0.1 Gy/day x 10 over a 12 day period).

5. We are testing the hypothesis that fractionated high-LET irradiation induces persistent inflammatory signaling that does not occur in single doses. An alternative hypothesis is that there could be a role of stem cell repopulation for lung tissue that is important in cancer progression. Thus, acute doses of high-LET ions may increase cell killing, reducing the number of target stem cells available for initiation and progression to lung cancer.

6. We have provided non lung tissues from our control and irradiated mice to other investigators (Ameila Eisch, brain tissue; Zhi-Min Yuan, kidney, liver, spleen)

Core A: Administrative Core. Leads: John Minna and Jerry Shay, and UTSW NSCOR

Administrator: Brenda Zielke.

The Administrative Core has established accounts and control over disbursement of all funds from this NSCOR application. It has also kept track of all personnel employed by this NSCOR. It has provided a central place for communication and coordinated data sharing In addition, it has established a series of biweekly to monthly scientific and planning meeting of the NSCOR investigators at UTSW. Approximately 25 scientific personnel attend each of these meetings. At each meeting administrative matters are discussed, experiments are planned, administrative changes made by the senior leadership team, and most importantly a scientific presentation by one of the NSCOR investigators is made and critiqued by the other investigators. Dr. Minna serves as the Administrative Core Lead with Dr. Jerry Shay as the Administrative Core Co-Lead. The Administrative Core coordinated all of the arrangements for the participation of several NSCOR investigators (Minna, Shay, Chen, Story, Roig, Burma, Delgado) in the 19th NSCOR Investigators Workshop held in Philadelphia, PA June 30-July 2, 2008. The Administrative core also arranged the Internal Scientific Advisory Board Review (March 4, 2008 as part of a mini-retreat at UT Southwestern) and the External Advisory Board Review (October 15, 2008 Mary Helen Barcellos-Hoff, Robert Ullrich and Martin Brown participating).

Core B. Cell Culture. Core Leads: John D. Minna, M.D. and Jerry Shay, Ph.D.

Enhancement of human bronchial epithelial cell 3D lung model of tissue like development There is increasing evidence that cancer arises from stem cell populations that are important for the normal turnover of tissues and the rapid repair of tissues in instances of injury. In the lung there is evidence for a bronchiole alveolar stem cell (BASC) population that may give rise to lung carcinoma. We developed a panel of immortalized human bronchial epithelial cells (HBECs) and have previously demonstrated that these cells when placed in an air-tissue culture medium interface overlying a fibroblast stromal collagen matrix could differentiate into mucous and ciliated bronchial epithelial cells. These results were expected since the cells originated from the central airways and were obtained from mid size bronchial tissues. We examined these cells for a variety of differentiated cell markers and observed that they expressed p63 (a transit amplifying cell marker) and surprisingly surfactant protein A (SP-A), CC10, a Clara cell marker, and when differentiated weak expression of the thyroid transcription factor TITF1 (known to be important in adenocarcinomas of the peripheral airway of the lungs). These peripheral markers associated with Clara and type II alveolar cells imply that our immortalized HBECs have enhanced plasticity and should be considered pluripotent lung stem cells. We next placed these cells into matrigel overlying a lung fibroblast (IMR90) feeder layer and observed that the cells could efficiently differentiate into spheroids or cyst like structures. Studies of these HBECs growing in matrigel and forming cysts and SACs indicated they started to express SP-A and morphologically resembled small bronchioles and alveolar sacs. When examined in the electron microscope, these cells also had lamellar bodies which is a hallmark of type II alveolar cells.

Core C. Genomics and Proteomics Core. Core Lead: Michael Story, Ph.D., Ching-Reng Yang, Ph.D. Core Co-Lead.

Genomics Core: The Core has made two advances in technology that should be described. The first was the analysis of miRNA in the HBEC series. The data collected to date are unique to the radiation community at large. The second area of progress has to do with the generation by Illumina Corp of a new gene expression platform that will drive the price of expression analysis down by half. We are implementing this new platform. This is the second 50% reduction in costs associated with gene expression during the life of this current application. This has already allowed the incorporation of new analytic techniques, miRNA analysis, and with the second cost reduction should allow us the flexibility to expand our analysis further or into new areas of genomics and proteomics.

A. Specific Functions of the Genomics Core

Microarray expression profiling, genome wide DNA copy number (array CGH), and array based protein expression technologies are maturing. However, they are very precise technologies that requires extensive experience to perform and sophisticated instrumentation to carry out. Furthermore, having a single facility through which all array analysis passes minimizes the variability brought about by different individuals processing samples even though the same platform is being used. For these reasons we include a Core facility (Core C, Genomics and Proteomics) in this NSCOR grant. This NSCOR Core interfaces with the UTSW Cancer Center Microarray and the UTSW school wide Cores. The NSCOR Core has the following duties:

1. Interact with Project Leaders and investigators in the design of experiments that use microarray technologies (mRNA, miRNA, DNA copy number, and proteomics) in their experimental approach. Biostatistics and bioinformatics will also be integrated in this process.

2. Establish procedures for consistency in sample preparation and RNA, DNA, and protein extraction, test all RNA, DNA, and protein samples for integrity, and provide reports to that effect.

3. Perform all hybridizations, perform all scans, and place data on a secure server.

4. Provide early analysis on array quality control and preliminary data analysis to investigators.

5. Interact with the Biostatistics and Bioinformatics Core (Core D) to provide bioinformatic analysis of data according to project requirements.

NSCOR Core D: Biostatistics and Bioinformatics. Core Lead: Xian-Jin Xie, Ph.D.; Core Co-Lead: Luc Girard, Ph.D. and Yang Xie, Ph.D. In conjunction with the Genomics (Expression Profiling and Proteomics) Core (Core C) and the Cell Culture Core (Core B), the Biostatistics and Bioinformatics Core (Core D) has the following specific aims:

-To provide database support and expertise for the collection, storage, integration, and retrieval of all sorts of data generated from NSCOR projects.

-To provide valid and optimal statistical design and to conduct proper analysis required to address the specific aims of each project.

-To review background and rationale, to assist in the design, evaluation, and analysis of new research proposal arising from the individual projects.

-To assist the other Cores and Projects in the proper analysis and interpretations of different types of bioassays, particularly mega-data such as those generated from gene expression arrays.

-To assist in manuscript preparation and to review scientific submissions.

-To assist the NSCOR team in building radiation cancer risk models.

As we progress on all the NSCOR projects, significant amount of new data have been generated since our 2007 annual progress report. As a result of Core D effort, a comprehensive data management system has been established that allows efficient access and sharing of all NSCOR data. Data Collection Close to 10,000 samples (including cell pellets and cryogenically preserved cell lines) and experiments (irradiation, microarrays, mouse data) have so far been collected and stored in the NSCOR database. These data can be viewed or edited by any participating investigator provided they have the necessary permissions.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Delgado O, Richardson JA, Xie X-J, Story MD, Minna JD, Shay JW. "Invasive cancer induced by fractionated Fe particle radiation in the K-ras lung cancer susceptible mouse model." 19th Annual NASA Space Radiation Investigators' Workshop, Philadelphia, PA, July 2008.

19th Annual NASA Space Radiation Investigators' Workshop, Philadelphia, PA, July 2008. , Jul-2008

Abstracts for Journals and Proceedings Hoshiyama H, Kim J-Y, Wright WE, Shay JW. "Genome-wide human shRNA screening to dissect pathways involved in telomerase." EMBO (European Molecular Biology Organization) conference 2008, “Telomeres and the DNA damage response“ ; Villars-sur-Ollon, Switzerland, September 2008.

EMBO (European Molecular Biology Organization) conference 2008, “Telomeres and the DNA damage response“ ; Villars-sur-Ollon, Switzerland, September 2008. , Sep-2008

Abstracts for Journals and Proceedings Hazra S, Krysan K, Walser T, Gardner B, Shay JW, Minna JD, Horvarth S, Dubinett SM. "A systems approach to the pre-clinical evaluation of targeted chemoprevention for lung cancer." Seventh Annual AACR International Conference Frontiers in Cancer Prevention Research, Washington, DC, November 2008.

Seventh Annual AACR International Conference Frontiers in Cancer Prevention Research, Washington, DC, November 2008. , Nov-2008

Abstracts for Journals and Proceedings Shay JW. "Aging and Cancer: Are Telomeres and Telomerase the Connection?" Seventh Annual AACR International Conference Frontiers in Cancer Prevention Research, Washington, DC, November 2008.

Seventh Annual AACR International Conference Frontiers in Cancer Prevention Research, Washington, DC, November 2008. , Nov-2008

Abstracts for Journals and Proceedings Park S, Perez, VL, Sato M, Peyton M, Gazdar A, Minna JD, Story MD. "Comparison of cellular transformation frequencies after low and high LET radiation exposures in isogenic normal human bronchial epithelial cells." Presented at the 44th Annual Meeting of the Radiation Research Society, Boston, MA, September 2008.

44th Annual Meeting of the Radiation Research Society, Boston, MA, September 2008. , Sep-2008

Abstracts for Journals and Proceedings Minna JD, Ding L, Park S, Sato M, Yang C-R, Girard L, Xie Y, Xie X-J, Peyton M, Gao B, Delgado O, Burma S, Chen D, Shay J, Story M. "mRNA, DNA repair and premalignant cellular responses of human bronchial epithelial cells to HZE particle and y-radiation." 19th Annual NASA Space Radiation Investigators' Workshop, Philadelphia, PA, July 2008.

19th Annual NASA Space Radiation Investigators' Workshop, Philadelphia, PA, July 2008. , Jul-2008

Abstracts for Journals and Proceedings Xie XJ, Bian A. "Goodness-of-fit tests for ordinal response regression models with continuous covariates." 8th Annual Hawaii International Conference on Statistics, Mathematics and Related Fields, Honolulu, Hawaii, January 13-15, 2009.

8th Annual Hawaii International Conference on Statistics, Mathematics and Related Fields, Honolulu, Hawaii, January 13-15, 2009. , Jan-2009

Abstracts for Journals and Proceedings Xie Y, Ding L, Xiao G, Allen J, Story M. "Statistical methods to preprocess Illumina microarray data." Eastern North Atlantic Region (ENAR) of the International Biometric Society Workshop, Arlington, VA, March 16, 2008.

Eastern North Atlantic Region (ENAR) of the International Biometric Society Workshop, Arlington, VA, March 16, 2008. , Mar-2008

Abstracts for Journals and Proceedings Xie Y, Carlin B. "Bayesian learning and identifiability in Hierarchical Models." Bayesian Biostatistics Conference, MD Anderson Cancer Center, Houston, Texas, January 2008.

Bayesian Biostatistics Conference, MD Anderson Cancer Center, Houston, Texas, January 2008. , Jan-2008

Abstracts for Journals and Proceedings Asaithamby A, Yannone SM, Qin J, Chen DJ. "Werner syndrome protein is phosphorylated by DNA-dependent protein kinase catalytic subunit and regulates DNA double-strand break repair." ICRR meeting, Radiation Research Society, San Francisco, California, July 2007.

ICRR meeting, Radiation Research Society, San Francisco, California, July 2007. , Jul-2007

Abstracts for Journals and Proceedings Asaithamby A, Uematsu N, Chen DJ. "Visualization of HZE particles induced DNA double strand break formation and repair in living cells." 18th Annual NASA Space Radiation Investigators Workshop, Rohnert Park, California, July 13-16, 2007.

18th Annual NASA Space Radiation Investigators Workshop, Rohnert Park, California, July 13-16, 2007. , Jul-2007

Abstracts for Journals and Proceedings Shay JW. "Aging and cancer: Are telomeres and telomerase the connection? " 39th Annual Meeting of the EMS (Environmental Mutagen Society), Puerto Rico, October, 2008.

39th Annual Meeting of the EMS (Environmental Mutagen Society), Puerto Rico, October, 2008. , Oct-2008

Articles in Peer-reviewed Journals Roig AI, Rusek A, Hight SK, Minna JD, Shay JW, Story MD. "A biological Bragg Curve for accelerated Fe particles: DNA damage intensity in fibroblasts in a 3 dimensional collagen matrix correlates with the Bragg curve energy distribution of a high LET particle." Radiat Res, submitted, January 2009. , Jan-2009
Articles in Peer-reviewed Journals Xie Y, Wang X, Story MD. "Statistical Methods of Background Correction for Illumina BeadArray." Bioinformatics, 2009, (in press). , Jan-2009
Articles in Peer-reviewed Journals Uematsu N, Weterings E, Yano K, Morotomi-Yano K, Jakob B, Taucher-Scholz G, Mari PO, van Gent DC, Chen BP, Chen DJ. "Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks." J Cell Biol. 2007 Apr 23;177(2):219-29. PMID: 17438073 , Apr-2007
Articles in Peer-reviewed Journals Asaithamby A, Uematsu N, Chatterjee A, Story MD, Burma S, Chen DJ. "Repair of HZE-particle-induced DNA double-strand breaks in normal human fibroblasts." Radiat Res. 2008 Apr;169(4):437-46. PMID: 18363429 , Apr-2008
Articles in Peer-reviewed Journals Ding LH, Xie Y, Park S, Xiao G, Story MD. "Enhanced identification and biological validation of differential gene expression via Illumina whole-genome expression arrays through the use of the model-based background correction methodology." Nucleic Acids Res. 2008 Jun;36(10):e58. PMID: 18450815 , Jun-2008
Articles in Peer-reviewed Journals Mukherjee B, Camacho CV, Tomimatsu N, Miller J, Burma S. "Modulation of the DNA-damage response to HZE particles by shielding." DNA Repair (Amst). 2008 Oct 1;7(10):1717-30. PMID: 18672098 , Oct-2008
Articles in Peer-reviewed Journals Shay JW, Keith WN. "Targeting telomerase for cancer therapeutics." Br J Cancer. 2008 Feb 26;98(4):677-83. Review. PMID: 18231105 , Feb-2008
Articles in Peer-reviewed Journals Agarwal M, Pandita S, Hunt CR, Gupta A, Yue X, Khan S, Pandita RK, Pratt D, Shay JW, Taylor JS, Pandita TK. "Inhibition of telomerase activity enhances hyperthermia-mediated radiosensitization." Cancer Res. 2008 May 1;68(9):3370-8. PMID: 18451164 , May-2008
Articles in Peer-reviewed Journals Yano K, Chen DJ. "Live cell imaging of XLF and XRCC4 reveals a novel view of protein assembly in the non-homologous end-joining pathway." Cell Cycle. 2008 May 15;7(10):1321-5. PMID: 18418068 , May-2008
Articles in Peer-reviewed Journals Weterings E, Chen DJ. "The endless tale of non-homologous end-joining." Cell Res. 2008 Jan;18(1):114-24. Review. PMID: 18166980 , Jan-2008
Articles in Peer-reviewed Journals Roig AI, Hight SK, Shay JW. "Two- and three-dimensional models for risk assessment of radiation-enhanced colorectal tumorigenesis." Radiat Res. 2009 Jan;171(1):33-40. PMID: 19138051 , Jan-2009
Articles in Peer-reviewed Journals Gagos S, Chiourea M, Christodoulidou A, Apostolou E, Raftopoulou C, Deustch S, Jefford CE, Irminger-Finger I, Shay JW, Antonarakis SE. "Pericentromeric instability and spontaneous emergence of human neo-acrocentric and minute chromosomes in the alternative pathway of telomere lengthening." Cancer Res. 2008 Oct 1;68(19):8146-55. PMID: 18829574 , Oct-2008
Awards Asaithamby A. "Scholar in Training Travel Award to attend 2007 ICRR meeting held in San Francisco, by Radiation Research Society to Asaithamby, July 2007." Jul-2007
Awards Asaithamby A. "Travel Award to attend 18th Annual NASA Space Radiation Investigators' Workshop, Rohnert Park, California, July 13-16, 2007." Jul-2007
Awards Park S. "2008 NASA Space Radiation Summer School, July 2008." Jul-2008
Awards Camacho C. "008 NASA Space Radiation Summer School, July 2008." Jul-2008
Awards Camacho C. "Third prize in the poster contest at the 19th Annual NASA Space Radiation Investigators' Workshop, Philadelphia, PA, June 30-July 2, 2008." Jul-2008
Awards Gonzalez O. "2007 NASA Space Radiation Summer School, Brookhaven National Laboratory, June 2007." Jul-2007
Awards Park S. "Travel award to attend 44th Annual Meeting of the Radiation Research Society, Boston, MA, September 2008." Sep-2008
Books/Book Chapters Xie X-J. "Goodness-of-Fit Tests for Logistic Regression Models." VDM Verlag Publishing, Inc., 2008., Jan-2008
 
Fiscal Year: FY 2008  Task Last Updated:  01/29/2008 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 03/31/2005  End Date:  03/31/2010 
No. of Post Docs: No. of PhD Degrees: 
No. of PhD Candidates: No. of Master' Degrees:   
No. of Master's Candidates: No. of Bachelor's Degrees:   
No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor:   Contact Phone:   
Contact Email:  
Flight Program:  
Flight Assignment: NOTE: Start/end dates changed to 3/31/2005-3/31/2010 per grant documents from PI (12/06)

 

Key Personnel Changes/Previous PI: We have made several administrative changes during the past reporting period. In 2007 Yang Xie, Ph.D. joined the Biostatistical Bioinformatics Core D and provides special expertise in analysis of array expression profiling data. Cheng-Ring Yang, Ph.D. joined the Genomics and Proteomics Core (Core C) to develop and provide expertise on proteomics profiling. Mitsuo Sato, M.D., Ph.D. returned to a faculty position in Japan and his work in Projects 1 and 3 are being taken over by Asst. Professor of Pharmacology Boning Gao, Ph.D. who is also expert in the HBEC system that Dr. Sato had provided.  
COI Name (Institution): Burma, Sandeep   ( The University of Texas Southwestern Medical Center )
Chen, David   ( The University of Texas Southwestern Medical Center )
Gazdar, Adi   ( The University of Texas Southwestern Medical Center  )
Girard, Luc  ( The University of Texas Southwestern Medical Center  )
Richardson, James   ( The University of Texas Southwestern Medical Center  )
Shay, Jerry   ( The University of Texas Southwestern Medical Center  )
Story, Michael  ( The University of Texas Southwestern Medical Center  )
Wright, Woody  ( The University of Texas Southwestern Medical Center  )
Xie, Xian-Jin  ( The University of Texas Southwestern Medical Center )
Minna, John  ( The University of Texas Southwestern Medical Center )
Yang, Chin-Reng  ( The University of Texas Southwestern Medical Center ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multi-step pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes will also be compared in these models to that of gamma-irradiation. These dose risk assessments will measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. They will also measure effects on development of cancer “stem-like” cells (“cancer initiating cells”). 3D organotypic culture and animal models will also allow measurement and risk estimation in both tissue surrogates and in vivo tissue situation including measurement of “bystander” effects. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORS and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have preliminary studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we have initiated experiments testing whether several transgenic mouse models of lung cancer after low and high LET exposure increase the early onset of lung cancer in the whole animal in real time. The NSCOR has 4 Projects: 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation; 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells; 3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and 4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models. These projects are supported by 4 Cores: Administrative; Cell Culture; Genomics and Proteomics; and Biostatistics and Bioinformatics. In addition, with funding from the Department of Energy (DOE) contributing to this NSCOR, studies of low-LET and low dose rate radiation in the same model systems are also being undertaken. The HZE particle irradiation will be done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer, a Center for the NCI Early Detection Research Network (EDRN) program, and is part of the NCI sponsored Genetic Epidemiology of Lung Cancer Consortium (GELCC) there will be great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits: Lung cancer is the leading cause of cancer death in both women and men in the USA and in the Western world. While lung cancer is caused by smoking exposure in 85% of people, because of the large numbers the remaining 15% of cases arising in life time never smokers (~25,000 patients per year) is a sizable health problem in the USA. Likewise, 50% of all new lung cancer cases occur in former smokers (quitting over 5 years before). In all of these scenarios a major underlying question has been the role of other environmental carcinogens especially environmental radiation. One source of this is radon gas in the environment including radon gas exposure in homes with alpha particles (which represent high LET radiation). Thus, the interaction of radiation in the environment on Earth including from alpha particles in the genesis of lung cancer and quantitation of the effect of such radiation on lung epithelial cells is of major health importance. Radiation is a known carcinogenic influence but the molecular events in lung epithelial cells in response to radiation need to be determined. A key element of this is the impact of radiation on lung epithelial cells with preneoplastic lesions. Our NSCOR uses innovative technology of human bronchial epithelial cells (HBECs) developed by us (coming from 45 different individuals) which have also been engineered to have known oncogenic preneoplastic changes (such as oncogenic KRAS, knockdown of or expression of mutant p53, expression of oncogenic mutant EGFR, expression of c-myc). Our studies are providing quantitative data following high LET irradiation on these HBECs. The primary benefits from this work will be the development of quantitative risk assessment models for astronauts of high LET radiation in space. However, the results will also have major impact and benefit to life on Earth includes new knowledge of the effects on human lung epithelial cells of irradiation in terms of quantitative genetic and epigenetic changes and gene expression changes following irradiation which are of potential importance as markers of radiation exposure that could occur through environmental exposures, accidentally or through terrorism. As part of this will be the quantitative response of DNA repair pathways to radiation including specific DNA repair components such as DNA repair enzymes and signaling pathways in lung epithelial cells. We are also determining quantitative changes in biologic phenotypes of lung epithelial cells in response to radiation which include colony formation in liquid and soft agar (anchorage independent growth) and differentiation and invasion in 3 dimensional cultures, and finally growth in vivo as either xenografts (for human cells) or endogenous lung cancers (for mouse models of lung cancer). From all of the above information we will identify novel molecular biomarkers (mRNA, DNA, protein) for lung carcinogenesis. Finally, we are also learning about inter individual variation of the response of HBECs to radiation with respect to the various genetic, epigenetic, mRNA and protein expression, and signaling pathway changes. Our large HBEC panel coupled with new high density single nucleotide polymorphism (SNP) analysis will allow describing polymorphic differences in such responses.

 

Task Progress: Project 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation.

Specific Aim 1: To quantitate the effects of HZE particle irradiation in causing genetic changes in HBECs and their isogenic derivates.

Specific Aim 2. To quantitate the amount of epigenetic changes caused by HZE particle irradiation in HBECs and their isogenic derivates using high throughput quantitative methylation specific PCR (MSP) assays for acquired promoter methylation

Specific Aim 3. To quantitate the effects of HZE particle irradiation in causing expression profile (mRNA and protein) changes in human bronchial epithelial cells (HBECs) and their isogenic derivates containing various lung cancer-related mutations using array based genome wide and multi protein detecting approaches.

Specific Aim 4. To determine the effect of inter-individual variation on the effects of HZE particle irradiation in causing genetic changes, epigenetic changes, mRNA and protein expression profile changes in human bronchial epithelial cells (HBECs).

We continued studies of gene expression changes following various types of radiation outlined in Aim 3. mRNA gene expression profiles were studied with Illumina v2 arrays before and at different time points after radiation. Supervised clustering methods identified samples by the radiation received via their gene response over time. We also examined kinetic differences rather than static differences as described above. The overall pattern of expression of genes was used to develop predictors of radiation response. From 1480 genes, temporal patterns were developed and 322 identified 12 different patterns

Project 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells.

Aim 1. To test the hypothesis that the immediate cellular response to DNA damage induced by HZE would be different and more complex than that induced by X-rays.

Aim 2. To test the hypothesis that persistent DNA damage induced by HZE is more complex and difficult to repair than that induced by X-rays and, therefore, more deleterious.

Aim 3. To determine if damage induced by HZE is preferentially repaired by non homologous end joining (NHEJ) or homologous recombination (HR) or is preferentially repaired in certain phases of the cell cycle.

Aim 4. To compare the response of human bronchial epithelial cells as non immortalized, immortalized, and with premalignant oncogenic changes in 3D cultures to HZE exposure using a high throughput approach.

Both X-rays and 1 GeV Fe particles elicit similar responses at the sites of DNA breaks. However, Fe-induced damage is repaired more slowly and a large fraction of breaks are not rejoined, thereby resulting in the higher relative biologic effect (RBE) of HZE particles. Using charged particles of increasing molecular weights (O vs. Si vs. Fe) we find that the incidence of un-rejoined breaks increases with increasing particle molecular weight presumably due to increasing complexity of the damage induced. We find that the capacity of both lung fibroblasts and HBECS to repair HZE-induced DNA damage is severely limited compared to gamma rays No major differences are seen between 2D and 3D cultures for fibroblasts. The capacity of HBECs to repair gamma-ray induced DNA damage varies depending upon the cancer-promoting oncogenic changes they harbor – these differences are minimized though in response to 1 GeV Fe.

Project 3. Effect of HZE Particle Irradiation on Functional Progression of Human Lung Cancer

Specific Aim 1. Effects of HZE particles on early events in lung cancer progression (soft agar colony formation and other markers of cellular transformation).

Specific Aim 2. Effects of HZE particles on intermediate events in lung cancer progression (invasion in 3 D organotypic cultures).

Specific Aim 3. Effects of HZE particles on late events in lung cancer progression (ability to form tumors in immune deprived mice).

Soft agar colony forming ability increases for both KRAS+p53 lines as a function of time and dose, and that their response is much greater than for HBEC3-KT (without oncogenic preneoplastic changes). There is no radiation-induced increase in soft agar colony forming ability in cells that over-express either wild type or mutant EGFR. Using quantiative methylation specific PCR (MSP) four of eleven markers found to be methylated are RASSF1A, APC, DcR1, and HCAD with a trend of methylation increasing with time after radiation treatment. We are testing the HZE particle and low LET irradiated HBEC cells of various genotypes for their ability to form tumors in immunodeprived mice as a test of the last events to give full tumor progression. We are also determining the effect of irradiation on the number of cancer stem cell like cells (cancer forming cells) using new sphere forming assays and quantitative mRNA expression profiling for a panel of ~30 stem cell genes.

Project 4. Effects of HZE Particles on the Development of Lung Cancer in Novel Mouse Models

Specific Aim 1. Use K-ras transgenic mouse model of human lung cancer to determine effects of low- and high-LET radiation in both living animals and in isolated tissues.

Throughout the second year of the UTSW NSCOR, experimental groups of approximately one hundred LA1 K-ras animals each were established and irradiated. Two groups were shipped to Brookhaven National Laboratory (BNL) on Long Island, NY for the NSRL-06A and NSRL-06C 56Fe high-LET radiation runs and respectively irradiated with a fractionated or single dose of 1.0 Gy 56Fe particles. Another two experimental groups were irradiated at UTSW with either a fractionated or single dose of 1.0 Gy X-rays for analysis of the effects of low-LET radiation. A fifth group of LA1 K-ras animals was left unirradiated for comparison to all irradiation groups. Irradiation of wild type littermates with a fractionated dose but not a single dose of 1.0 Gy 56Fe particles significantly decreases their survival compared to unirradiated wild-type animals. Mutant animals irradiated with either a fractionated or single dose of 1.0 Gy 56Fe particles have significantly decreased lifespan compared to unirradiated mutant LA1 K-ras mice. In comparison, irradiation with 1.0 Gy X-rays does not affect the survival of mutant or wild type littermate LA1 K-ras mice regardless of administration method. Radiation does not appear to affect overall tumor incidence in irradiated mutant LA1 K-ras mice compared to unirradiated mutant controls. There is a dramatic progression in the grade of lesions compared to unirradiated mutant mice, X-ray irradiated mutant mice, or mutants irradiated with a single 1.0 Gy dose of 56Fe particles. The number adenocarcinomas are significantly increased which display highly aggressive and invasive characteristics of malignancy, which more closely mimics clinical features of lung cancer in humans. Whole genome mRNA analysis was performed and unsupervised cluster analysis of resultant gene expression profiles has suggested divergent responses between mutant and wild type littermate lung associated with radiation type.

Specific Aim 2. Generate or obtain other transgenic mice susceptible to lung cancer to determine the effects of low- and high-LET radiation in both living animals and in isolated tissues. These include the mTERT knockout mouse and the EGFR TK domain mutation mouse. We will initiate pilot experiments of the mTERT KO mouse during the next review period to determine if more end joining, end-end fusions, cytogenetic alterations, micronuclei etc occur when these mice are exposed to 1 and 3 Gy of X-ray. The EGFR mouse model we have obtained has a EGFR mutation that can be induced by tetracycline in the food or drinking water.

Core B. Cell Culture

We placed HBECs into matrigel overlying a lung fibroblast (IMR90) feeder layer and observed that the cells could efficiently differentiate into spheroids or cyst like structures that express SP-A and morphologically resembled small bronchioles and alveolar sacs, and also had lamellar bodies which is a hallmark of type II alveolar cells.

Core C. Genomics and Proteomics Core

This core has started to develop reverse Phase Protein Lysate Arrays (RPPAs). This will allow us to examine the expression of one to several proteins in a moderate throughput platform (hundreds of samples simultaneously). We have also perfected the use of comparative genomic hybridization arrays (aCGH). This will allow us to examine chromosome copy variation which is likely to be important for the analysis of cancer progression studies being carried out in Project 3.

Core D: Biostatistics and Bioinformatics

A centralized database with a server-client architecture has been created to store and share data generated with the various NSCOR experiments. This multi-user database system has already been implemented and extensively tested in other projects and is being adapted and expanded more specifically for NSCOR. The database allows easy access and sharing of all NSCOR data by all investigators. Thousands of collected samples and large sets of data can be organized and shared across investigators and statisticians. Its centralized nature makes it possible to schedule regular backups, to ensure that data are organized in a way that analysis and data interpretation are facilitated, and to allow investigators to keep track of experiments and samples and thereby substantially minimize errors. The database is accessible through the Internet making it possible to enter data in real-time from remote sites such as BNL.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Park S, Sato M, Peyton M, Minna JD, Story MD. "Determination of radiation survival, cellular transformation, and tumorigenicity in isogenic human bronchial epithelial cells after exposure to low and high LET radiations." 18th NASA investigator’s annual meeting, Monterey , CA, July 13-16, 2007.

18th NASA investigator’s annual meeting, Monterey , CA, July 13-16, 2007. , Jul-2007

Abstracts for Journals and Proceedings Ding LH, Park S, Minna JD, Story MD. "Comparison of gene expression profiles of human bronchial epithelial cells after HZE and gamma-ray radiation." 18th NASA investigator’s annual meeting, Monterey/Rohnert Park, CA, July 13-16, 2007.

18th NASA investigator’s annual meeting, Monterey/Rohnert Park, CA, July 13-16, 2007. , Jul-2007

Abstracts for Journals and Proceedings Delgado O, Minna DG, Richardson JA, Xie X, Ding L, Story MD, Minna JD, Shay JW. "Space radiation effects on lung cancer progression in LA1 K-ras mouse model of lung cancer." 18th Annual NASA Space Radiation Investigators Workshop, Rohnert Park, California, July 13-16, 2007.

18th Annual NASA Space Radiation Investigators Workshop, July, 2007. , Jul-2007

Abstracts for Journals and Proceedings Minna JD, Sato M, Girard L, Xie X-J, Yang C-R, Peyton M, Sheridan S, Burma S, Chen D, Shay J, Story M. "mRNA, DNA repair and premalignant cellular responses of human bronchial epithelial cells to HZE particle and gamma-radiation." 18th Annual NASA Space Radiation Investigators Workshop, Rohnert Park, California, July 13-16, 2007.

18th Annual NASA Space Radiation Investigators Workshop, July, 2007. , Jul-2007

Abstracts for Journals and Proceedings Shay JW. "Is Cellular Ageing Due to DNA Damage Signaling from Short Telomeres, Telomere Position Effects or Both?" Warsaw Aging Symposium, Seneca, Warsaw Poland, October 3-7, 2007.

Warsaw Aging Symposium, Seneca, Warsaw Poland, October 3-7, 2007. , Oct-2007

Abstracts for Journals and Proceedings Shay JW, Sato M, Lee W, Girard L, Xie XJ, Xie Y, Sullivan J, Spinola M, Minna JD. "Telomerase immortalized human bronchial epithelial cells (HBECs) have stem cell characteristics." ISLAC International Congress, Seol Korea, Sept 1-6, 2006.

ISLAC International Congress, Seol Korea, Sept 1-6, 2006. , Sep-2006

Abstracts for Journals and Proceedings Shay JW. "Title not available." Jack Gross Memorial Lecture, Jerusalem, Israel, March 24-28, 2006.

Jack Gross Memorial Lecture, Jerusalem, Israel, March 24-28, 2006. , Mar-2006

Abstracts for Journals and Proceedings Shay JW. "Title not available." Beatson International Cancer Conference, Glasgow, Scotland, June 17-20, 2006.

Beatson International Cancer Conference, Glasgow, Scotland, June 17-20, 2006. , Jun-2006

Abstracts for Journals and Proceedings Shay JW. "Title not available." Distinguished Visiting Professor Pathology Grand Round, Johns Hopkins, November 4, 2006.

Distinguished Visiting Professor Pathology Grand Round, Johns Hopkins, November 4, 2006. , Nov-2006

Abstracts for Journals and Proceedings Minna JD. "Title not available." Visiting Professor Lecture, Yale University Medical Center, February 2006.

Visiting Professor Lecture, Yale University Medical Center, February 2006. , Feb-2006

Abstracts for Journals and Proceedings Minna JD. "Title not available." Visiting Professor Lecture, Vanderbilt University Medical Center, April 2006.

Visiting Professor Lecture, Vanderbilt University Medical Center, April 2006. , Apr-2006

Abstracts for Journals and Proceedings Minna JD. "Title not available." Visiting Professor, Lecture, M.D. Anderson Cancer Center, May 2006.

Visiting Professor, Lecture, M.D. Anderson Cancer Center, May 2006. , May-2006

Abstracts for Journals and Proceedings Shay JW. "Title not available." 16th Annual Space Radiation Health Investigator's Workshop, Port Jefferson, Long Island, NY, May 15-18, 2005.

16th Annual Space Radiation Health Investigator's Workshop, Port Jefferson, Long Island, NY, May 15-18, 2005. , May-2005

Abstracts for Journals and Proceedings Shay JW. "DNA Damage Signaling and Aging: Relationships to Telomeres and Telomerase." 14th International Symposium on Microdosimetry – Venice, Italy, November 13-18, 2005.

14th International Symposium on Microdosimetry – Venice, Italy, November 13-18, 2005. , Nov-2005

Abstracts for Journals and Proceedings Minna JD, Sato M, Sheridan S, Giraard L, Gazdar AF, Chen D, Story MD, Shay JW. "Effect of HZE Particle Irradiation on Progressing Human Bronchial Epithelial Cells Towards Malignancy." 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop. Moscow – St. Petersburg, June 5 - 9, 2006.

4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop. Moscow – St. Petersburg, June 5 - 9, 2006. , Jun-2006

Abstracts for Journals and Proceedings Delgado O, Roig A, Story MD, Richardson J, Minna JD, Shay JW. "Effects of Low and High LET Irradiation Exposure on the Development of Lung Cancer in Novel Mouse Models." 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop, Moscow – St. Petersburg, June 5 - 9, 2006.

4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop, Moscow – St. Petersburg, June 5 - 9, 2006. , Jun-2006

Abstracts for Journals and Proceedings Story MD, Minna JD, Sheridan S, Ding LH, Girard L, Chen DJ, Shay JW. "Effects of Low- and High-LET Radiation Exposures in Human Bronchial Epithelial Cells." 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop, Moscow – St. Petersburg, June 5 - 9, 2006.

4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators’ Workshop, Moscow – St. Petersburg, June 5 - 9, 2006. , Jun-2006

Abstracts for Journals and Proceedings Story MD, Ding LH, Park S, Yang CR, Sato M, Girard L, Xie Y, Xie XJ, Peyton M, Gao B, Burma S, Chen,D, Shay JW, Minna JD. "mRNA and Premalignant Cellular Responses of Human Bronchial Epithelial Cells to low dose gamma-Radiation." DOE VII Low Dose Program Investigators’ Workshop, Washington, DC, Jan 21-23, 2008.

DOE VII Low Dose Program Investigators’ Workshop, Washington, DC, Jan 21-23, 2008. , Jan-2008

Abstracts for Journals and Proceedings Minna JD, Ding L, Park S, Sato M, Yang CR, Girard L, Xie Y, Xie XJ, Peyton M, Gao B, Burma S, Chen D, Shay JW, Story MD. "mRNA, DNA Repair and Premalignant Cellular Responses of Human Bronchial Epithelial Cells to HZE Particle and gamma-Radiation." NASA Human Research Program Investigators’ Workshop, League City Texas, Feb 3-6, 2008.

NASA Human Research Program Investigators’ Workshop, League City Texas, Feb 3-6, 2008. , Feb-2008

Articles in Peer-reviewed Journals Pearce VP, Sherrell J, Lou Z, Kopelovich L, Wright WE, Shay JW. "Immortalization of epithelial progenitor cells mediated by resveratrol." Oncogene. 2007 Oct 29; [Epub ahead of print] PMID: 17968319 , Oct-2007
Articles in Peer-reviewed Journals Asaithamby A, Uematsu N, Chatterjee A, Story MD, Burma S, Chen DJ. "HZE particle-induced DNA double-strand break repair in normal human fibroblasts." Radiation Research 2008 (In Press). , Jan-2008
Articles in Peer-reviewed Journals Ding L-H, Xie Y, Park S, Xiao G, Story MD. "Enhanced identification and biological validation of differential gene expression via Illumina whole genome expression arrays through the use of the Model Based Background Correction Methodology." Nucleic Acids Research (submitted, pending revisions), January 2008. , Jan-2008
Articles in Peer-reviewed Journals Sato M, Vaughan MB, Girard L, Peyton M, Lee W, Shames DS, Ramirez RD, Sunaga N, Gazdar AF, Shay JW, Minna JD. "Multiple oncogenic changes (K-rasV12, p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells." Cancer Res. 2006 Feb 15;66(4):2116-28. PMID: 16489012 , Feb-2006
Articles in Peer-reviewed Journals Shay JW, Wright WE. "Telomerase therapeutics for cancer: challenges and new directions." Nat Rev Drug Discov. 2006 Jul;5(7):577-84. Review. PMID: 16773071 , Jul-2006
Articles in Peer-reviewed Journals Das AK, Sato M, Story MD, Peyton M, Graves R, Redpath S, Girard L, Gazdar AF, Shay JW, Minna JD, Nirodi CS. "Non-small cell lung cancers with kinase domain mutations in the epidermal growth factor receptor are sensitive to ionizing radiation." Cancer Res. 2006 Oct 1;66(19):9601-8. PMID: 17018617 , Oct-2006
Articles in Peer-reviewed Journals Vaughan MB, Ramirez RD, Wright WE, Minna JD, Shay JW. "A three-dimensional model of differentiation of immortalized human bronchial epithelial cells. " Differentiation. 2006 Apr;74(4):141-8. PMID: 16683984 , Apr-2006
Articles in Peer-reviewed Journals Shames DS, Girard L, Gao B, Sato M, Lewis CM, Shivapurkar N, Jiang A, Perou CM, Kim YH, Pollack JR, Fong KM, Lam CL, Wong M, Shyr Y, Nanda R, Olopade OI, Gerald W, Euhus DM, Shay JW, Gazdar AF, Minna JD. "A genome-wide screen for promoter methylation in lung cancer identifies novel methylation markers for multiple malignancies." PLoS Med. 2006 Dec;3(12):e486. PMID: 17194187 , Dec-2006
Articles in Peer-reviewed Journals Jackson SR, Zhu CH, Paulson V, Watkins L, Dikmen ZG, Gryaznov SM, Wright WE, Shay JW. "Antiadhesive effects of GRN163L--an oligonucleotide N3'->P5' thio-phosphoramidate targeting telomerase." Cancer Res. 2007 Feb 1;67(3):1121-9. PMID: 17283146 , Feb-2007
Articles in Peer-reviewed Journals Das AK, Chen BP, Story MD, Sato M, Minna JD, Chen DJ, Nirodi CS. "Somatic mutations in the tyrosine kinase domain of epidermal growth factor receptor (EGFR) abrogate EGFR-mediated radioprotection in non-small cell lung carcinoma." Cancer Res. 2007 Jun 1;67(11):5267-74. PMID: 17545606 , Jun-2007
Articles in Peer-reviewed Journals Tsakiri KD, Cronkhite JT, Kuan PJ, Xing C, Raghu G, Weissler JC, Rosenblatt RL, Shay JW, Garcia CK. "Adult-onset pulmonary fibrosis caused by mutations in telomerase." Proc Natl Acad Sci U S A. 2007 May 1;104(18):7552-7. PMID: 17460043 , May-2007
Articles in Peer-reviewed Journals Shay JW, Wright WE. "Hallmarks of telomeres in ageing research." J Pathol. 2007 Jan;211(2):114-23. Review. PMID: 17200948 , Jan-2007
Articles in Peer-reviewed Journals Garcia CK, Wright WE, Shay JW. "Human diseases of telomerase dysfunction: insights into tissue aging." Nucleic Acids Res. 2007;35(22):7406-16. Review. PMID: 17913752 , Dec-2007
Articles in Peer-reviewed Journals Uematsu N, Weterings E, Yano K, Morotomi-Yano K, Jakob B, Taucher-Scholz G, Mari PO, van Gent DC, Chen BP, Chen DJ. "Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks." J Cell Biol. 2007 Apr 23;177(2):219-29. PMID: 17438073 , Apr-2007
Awards Delgado O, Minna DG, Richardson JA, Xie XJ, Ding LH, Story MD, Minna JD, Shay JW. "First Place Poster Award to Oscar Gonzales, Graduate Student, for: Space Radiation Effects on Lung Cancer Progression in LA1 K-ras Mouse Model of Lung Cancer, 18th NASA Investigators Meeting, Rohnert Park, CA July, 2007." Jul-2007
Awards Roig AI, Shay JW. "First Place Poster Award to Andres Roig, Postdoctoral Trainee, for: 2D and 3D Models for Risk Assessment of Space Radiation-Enhanced Colon Tumorigenesis, 18th NASA Annual Meeting, Rohnert Park, CA, July 2007." Jul-2007
Awards Peyton M, Gonzales OR. "2007 NASA Space Radiation Summer School, June 2007." Jun-2007
Books/Book Chapters Sato M, Shames DS, Girard L, Gazdar AF, Minna JD. "Molecular Genetics of Lung Cancer." in "The Molecular Basis of Cancer. 3rd ed." Ed. J. Mendelsohn, M.A. Israel, P.M. Howley, J.W. Gray, C.B. Thompson. Philadelphia : Saunders/Elsevier Press, 2008. p. (In Press). Expected Release Date: 03/28/2008., Jan-2008
Papers from Meeting Proceedings Shay JW. "Keynote Speaker, Genomic Instability and Cancer." Srinagar, Kashmir India, July 19-25, 2006.

Srinagar, Kashmir India, July 19-25, 2006. , Jul-2006

 
Fiscal Year: FY 2007  Task Last Updated:  01/30/2007 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 03/31/2005  End Date:  03/31/2010 
No. of Post Docs: No. of PhD Degrees: 
No. of PhD Candidates: No. of Master' Degrees:   
No. of Master's Candidates: No. of Bachelor's Degrees:   
No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor:   Contact Phone:   
Contact Email:  
Flight Program:  
Flight Assignment: NOTE: Start/end dates changed to 3/31/2005-3/31/2010 per grant documents from PI (12/06)

 

Key Personnel Changes/Previous PI: 12/2006: Lead of Core D - Bioinformatics and Biostatistics was changed to Dr. Xian-Jin Xie, when Dr. Raul Caetano was named dean of UT Southwestern Allied Health Sciences School and Dr. Harris assumed other responsibilties. Dr. Ruben Ramirez and Dr. Jonathan Abrams are no longer affiliated with this project. Their effort has been absorbed by personnel already supported by the project. They will not be replaced. Also because of his active role in all aspects of the NSCOR leadership, Dr. Jerry Shay has assumed the co-Lead role for the NSCOR Administrative Core for Dr. David Chen. 
COI Name (Institution): Burma, Sandeep   ( The University of Texas Southwestern Medical Center )
Chen, David   ( The University of Texas Southwestern Medical Center )
Gazdar, Adi   ( The University of Texas Southwestern Medical Center  )
Girard, Luc  ( The University of Texas Southwestern Medical Center  )
Richardson, James   ( The University of Texas Southwestern Medical Center  )
Shay, Jerry   ( The University of Texas Southwestern Medical Center  )
Story, Michael  ( The University of Texas Southwestern Medical Center  )
Wright, Woody  ( The University of Texas Southwestern Medical Center  )
Xie, Xian-Jin  ( The University of Texas Southwestern Medical Center )
Minna, John D. ( The University of Texas Southwestern Medical Center ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR proposal focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multistep pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes will also be compared in these models to that of X-irradiation. These dose risk assessments will measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. 3D organotypic culture and animal models will also allow measurement and risk estimation of “bystander” effects. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORS and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have preliminary studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we have developed plans for novel transgenic mouse models to monitor HZE particle induction of lung cancer in the whole animal in real time. The NSCOR has 4 Projects: 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation.; 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells; 3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and 4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models. These projects are supported by 4 Cores: Administrative; Cell Culture; Expression Profiling and Proteomics; and Biostatistics and Bioinformatics. In addition, with funding from the Department of Energy (DOE) contributing to this NSCOR, studies of low-LET and low dose rate radiation in the same model systems are also being undertaken. The HZE particle irradiation will be done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, and holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer and a Center for the NCI Early Detection Research Network (EDRN) program, there will be great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits: Lung cancer is the leading cause of cancer death in both women and men in the USA and in the Western world. While lung cancer is caused by smoking exposure in 85% of people, because of the large numbers the remaining 15% of cases arising in life time never smokers (~25,000 patients per year) is a sizable health problem in the USA. Likewise, 50% of all new lung cancer cases occur in former smokers (quitting over 5 years before). In all of these scenarios a major underlying question has been the role of other environmental carcinogens especially environmental radiation. One source of this is radon gas in the environment including radon gas exposure in homes with alpha particles (which represent high LET radiation). Thus, the interaction of radiation in the environment on Earth including from alpha particles in the genesis of lung cancer and quantitation of the effect of such radiation on lung epithelial cells is of major health importance. Radiation is a known carcinogenic influence but the molecular events in lung epithelial cells in response to radiation need to be determined. A key element of this is the impact of radiation on lung epithelial cells with preneoplastic lesions. Our NSCOR uses innovative technology of human bronchial epithelial cells (HBECs) developed by us (coming from 36 individuals) which have also been engineered to have known oncogenic preneoplastic changes (such as oncogenic KRAS and knockdown of p53). Our studies are providing quantitative data following high LET irradiation on these HBECs. The primary benefits from this work will be the development of quantitative risk assessment models for astronauts of high LET radiation in space. However, the results will also have major impact and benefit to life on Earth includes new knowledge of the effects on human lung epithelial cells of irradiation in terms of quantitative genetic and epigenetic changes and gene expression changes following irradiation. As part of this will be the quantitative response of DNA repair pathways to radiation including specific DNA repair components such as DNA repair enzymes and signaling pathways in lung epithelial cells. We are also determining quantitative changes in biologic phenotypes of lung epithelial cells in response to radiation which include colony formation in liquid and soft agar (anchorage independent growth) and differentiation and invasion in 3 dimensional cultures, and finally growth in vivo as either xenografts (for human cells) or endogenous lung cancers (for mouse models of lung cancer). From all of the above information we will identify novel molecular biomarkers (mRNA, DNA, protein) for lung carcinogenesis. Finally, we are learning about inter individual variation of the response of HBECs to radiation with respect to the various genetic, epigenetic, mRNA and protein expression, and signaling pathway changes.

 

Task Progress: Project 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation. Michael Story, Ph.D. Lead, Adi F. Gazdar, M.D. Co-Lead, John D. Minna, M.D. Co-Lead

The overall goals of project 1 are to establish the low LET and the HZE radio-response of human bronchial epithelial cells (HBECs) that have been genetically manipulated, with a variety of oncogenic changes that are often seen during the oncogenic progression towards lung carcinogenesis. These changes include: down regulation of p53; oncogenic KRASV12; and over-expression of wild type and mutant EGFR. These cell lines represent in vitro equivalents of preneoplastic lesions and even with multiple changes, these manipulated HBECs show only part of the malignant phenotype and are not capable of forming tumors in immunodeprived mice. We are performing standard survival analysis following radiation exposure, mRNA gene expression analysis, protein expression, DNA methylation, clonogenicity in soft agar (Project 3), and tumorigenicity (Project 3). We follow irradiated cell cultures up to 4 months post-irradiation in order to examine these endpoints in surviving cells in order to determine the effect the radiation exposure has on the oncogenic process. For low LET radiation exposures the radio-response of these cell lines was not uniform with mutant EGFR containing cells being radiosensitive. Cells exposed to Fe particles at 1000MeV (1 GeV Fe) are highly susceptible to killing, Si particle exposure at the same energy as the Fe particles, produced an intermediate killing effect, while H exposures showed a similar response as that of gamma-radiation. These results correspond well with the ionization intensity differences seen via DNA foci formation in HZE irradiated cells as determined by Project 2. The cells were collected at various time points following irradiation, the RNA was extracted and Illumina expression microarrays were performed. Acute changes (<24 h) were found in gene expression following exposure of human bronchial epithelial cells to irradiation. These changes were distinct for the various types of irradiation (Fe, Si, gamma) as well as for the various HBEC variants. Most experiments showed no long-term gene expression changes as an effect of irradiation.

Methylation of the promoter region of genes is a fundamental process by which tumor cells inactivate tumor suppressor genes. Our studies have focused on the development of methylation following irradiation of human bronchial epithelial cells (HBECs which are devoid of methylation of multiple genes frequently methylated and silenced in lung cancers. We studied the methylation status of 5 genes frequently methylated in lung cancers – RASSF1A, APC, CYT, GRB38 and RAR-beta. Methylation was measured using a TaqMan semi-quantitative real time PCR method. For all of the unmodified HBEC cell groups and for two of the modified HBECKTR53 cell groups, there were no significant differences between radiated and control groups. However for HBECKRASP53 cell group irradiated with HZE particles, there were important differences where the irradiated group which showed methylation of all 5 genes, and a progressive increase in methylation between the 7 and 14 day time points.

Project 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells. David Chen, Ph.D. Project Lead, Sandeep Burma, Ph.D. Co-Lead

With the ultimate objective of estimating cancer risks to humans from HZE particles, in this project we investigate the DNA damage responses of human cells to HZE, the link between these responses and cancer predisposition being very well established. We will take advantage of recent advances in the study of molecular mechanisms of DNA damage sensing, damage signaling, and DNA repair following low LET radiation to verify the hypothesis that HZE particles generate a different and/or unique radiation response in mammalian cells as compared to X-rays. Again we used the panel of isogenic HBEC cells with various oncogenic manipulations described above. Using 1 GeV Fe and X-rays to begin with we have worked out the early damage-responses, damage-repair kinetics, and persistence of DNA damage in primary human skin fibroblasts (HSFs) in great detail. Very briefly, both X-rays and 1 GeV Fe particles elicit similar responses at the sites of DNA breaks. However, Fe-induced damage is repaired more slowly and a large fraction of breaks are not rejoined, thereby resulting in the higher relative biologic effect (RBE) of HZE particles. Using charged particles of increasing molecular weights (O vs. Si vs. Fe) we find that the incidence of unrejoined breaks increases with increasing particle molecular weight presumably due to increasing complexity of the damage induced. Detailed time courses have already been carried out with a panel of HBECs irradiated with 1 GeV Fe particles. The resulting data are currently undergoing analyses.

Project 3. Effect of HZE Particle Irradiation on Functional Progression of Human Lung Cancer. John D. Minna, M.D. Project Lead, Jerry Shay, Ph.D. Co-Lead

Early Events. To evaluate the biologic effects of HZE particle irradiation on HBECs we performed soft agar colony formation assays. Isogenic HBECs with different genetic manipulations were irradiated with HZE and subcultured up to four months after the irradiation. Every month, soft agar colony formation assays were done. The experiments were repeated twice. Most of the oncogenic manipulations reproducibly enhanced the colony formation ability of HBECs without radiation exposure. However, no significant difference was seen between control and HZE-treated HBECs with the one exception of HBECKTR53 (with KRASV12 and p53 knockdown), where HZE irradiated HBECs formed more colonies than non-treated cells at all four time points. This was particularly apparent after correcting for radiation induced decrease in cell survival.

Late Events. We are testing the HZE particle and low LET irradiated HBEC cells of various genotypes for their ability to form tumors in immunodeprived mice as a test of the last events to give full tumor progression. As part of this we are also determining the effect of irradiation on the number of stem cell like cells using new sphere forming assays and quantitative mRNA expression profiling for a panel of ~20 stem cell proliferation program genes.

Project 4. Effects of HZE Particles on the Development of Lung Cancer in Novel Mouse Models. Jerry Shay, Ph.D. Project Lead, James Richardson, DVM, Ph.D.

Transgenic mouse models of human lung cancer have the advantage of a similar tumor development pathway compared to humans but with a shorter developmental period. The LA1 K-ras mouse model we have used contains a latent oncogenic K-ras allele integrated in the endogenous K-ras locus. The activation of the oncogenic allele occurs spontaneously preferentially in the lungs as the animals mature. Every animal containing the latent allele develops alveolar type II lineage lung cancer by the age of four to nine months and all animals die at approximately one year from tumor burden. Our primary objective is to determine if these animals, after low and high LET exposure, initiate or progress to cancer at increased frequency. Our long-term goal is to determine if there is an increase, decrease or no change of onset of death after HZE irradiation and ultimately to use this information for modeling risks in humans. Other end points include other cancer types, DNA damage and repair (gamma-H2AX, DNA-PKc) to correlate with the 2D and 3D cell culture experiments in projects 2 and 3. In addition, we will obtain histology and conduct expression profiles of low and high LET irradiated mouse lungs to compare to the cell culture experiments in Projects 1-3. We have conducted low LET (X-ray) at UT Southwestern Medical Center and high LET (Fe56) at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). 94 animals were irradiated with 0.2 Gy x 5 (1GeV Fe56), 97 animals were irradiated with 1.0 Gy (1GeV Fe56), 110 animals irradiated with 0.2 Gy x 5 (X-ray), and 100 animals irradiated with 1Gy (X-ray). In addition, we have 98 non-irradiated control animals. To examine acute responses (3 hr post fractionated and single dose low and high LET radiation), lung tissue was removed for pathologic examination and RNA isolated for gene expression arrays. Extracted RNA will be hybridized to Sentrix(R) Mouse-6 Expression BeadChips (Illumina). The first section was stained with Hematoxylin and Eosin for histopathological assessment and the following sections are currently being stained with representative markers of the DNA damage response such as gamma-H2AX and 53BP1. Kinetics in normal bronchus, normal alveolus, and tumor tissue will be evaluated separately to determine any differences. We are bringing on line a new mouse model of lung cancer using a transgenic mouse with a Tet-inducible mutant EGFR gene that results in lung cancer. There appears to be a slight decrease in the survival of the irradiated LA1 K-ras mutant and possibly wild-type animals compared to the unirradiated controls. There is also a strain background differences with the B6.1292 background being more susceptible to the radiation effects than the 129sv background. So far, irradiated mutant animals do not appear to have increased tumor incidence as determined by post-mortem evaluation. Overall these experiments allow us to study the effects of HZE and low LET radiation on the development of lung cancer in a whole animal model.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
Abstracts for Journals and Proceedings Story MD, Minna JD, Sheridan S, Ding L-H, Girard L, Chen DJ, Shay J. "Effects of Low- and High-LET Radiation Exposures in ..." 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop, Moscow - St. Petersburg Russia, June 5-9, 2006.

Abstracts, 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop, June 2006. , Jun-2006

Abstracts for Journals and Proceedings Minna JD, Sato M, Sheridan S, Girard L, Gazdar AF, Chen D, Story M, Shay J. "Effect of HZE Particle Irradiation on Progressing Human Bronchial Epithelial Cells Towards Malignancy." 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop Moscow - St. Petersburg Russia, June 5-9, 2006.

Abstracts, 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop, June 2006. , Jun-2006

Abstracts for Journals and Proceedings Delgado O, Roig A, Story M, Richardson J, Minna J, Shay J. "Effects of Low and High LET Irradiation Exposure on the Development of Lung Cancer in Novel Mouse Models" 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop Moscow - St. Petersburg Russia, June 5-9, 2006.

Abstracts, 4th International Workshop on Space Radiation Research and 17th Annual NASA Space Radiation Health Investigators' Workshop, June 2006. , Jun-2006

Articles in Peer-reviewed Journals Sato M, Vaughan MB, Girard L, Peyton M, Lee W, Shames DS, Ramirez RD, Sunaga N, Gazdar AF, Shay JW, Minna JD. "Multiple oncogenic changes (K-rasV12, p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells." Cancer Res. 2006 Feb 15;66(4):2116-28. PMID: 16489012 , Feb-2006
Articles in Peer-reviewed Journals Shay JW, Wright WE. "Telomerase therapeutics for cancer: challenges and new directions." Nat Rev Drug Discov. 2006 Jul;5(7):577-84. Review. PMID: 16773071 , Jul-2006
Articles in Peer-reviewed Journals Das AK, Sato M, Story MD, Peyton M, Graves R, Redpath S, Girard L, Gazdar AF, Shay JW, Minna JD, Nirodi CS. "Non-small cell lung cancers with kinase domain mutations in the epidermal growth factor receptor are sensitive to ionizing radiation." Cancer Res. 2006 Oct 1;66(19):9601-8. PMID: 17018617 , Oct-2006
Articles in Peer-reviewed Journals Vaughan MB, Ramirez RD, Wright WE, Minna JD, Shay JW. "A three-dimensional model of differentiation of immortalized human bronchial epithelial cells. " Differentiation. 2006 Apr;74(4):141-8. PMID: 16683984 , Apr-2006
Articles in Peer-reviewed Journals Jackson SR, Zhu C-H, Paulson V, Watkin L, Dikmen ZG, Gryaznov SM, Wright WE, Shay JW. "Anti-adhesive effects of GRN163L – an oligonucleotide, N3’-P5’ thio-phosphoramidate targeting telomerase." Cancer Res. (in press, 2007). , Jan-2007
Articles in Peer-reviewed Journals Uematsu, N, Weterings, E, Yano, K, Morotomi-Yano, K, Jakob, B, Taucher-Scholz, G, Mari, P, van Gent, DC, Chen, B, Chen, DJ. "Autophosphorylation of DNA-PKCS regulates its dynamics at DNA double-strand breaks." J Cell Biol, in revision, 2007. , Jan-2007
 
Fiscal Year: FY 2006  Task Last Updated:  01/31/2007 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 03/31/2005  End Date:  03/31/2010 
No. of Post Docs: No. of PhD Degrees:   
No. of PhD Candidates: No. of Master' Degrees:   
No. of Master's Candidates: No. of Bachelor's Degrees:   
No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor:   Contact Phone:   
Contact Email:  
Flight Program:  
Flight Assignment: NOTE: Start/end dates changed to 3/31/2005-3/31/2010 per grant documents from PI (12/06)

 

Key Personnel Changes/Previous PI: Dr. Story has become the Lead of Project #1 with Dr. Gazdar becoming the Co-Lead. This is because the majority of the findings appear to revolve about gene expression and genetic changes after HZE irradiation, and because of the large amount of HBEC cell survival studies that are needed that Dr. Story is leading. Dr. Minna and Dr. Shay have assumed responsibility for the Cell Culture Core. This occurred because of the focus and need for expanding human bronchial epithelial cells for nearly all NSCOR related experiments and their own labs expertise in this. Dr. Story has assumed responsibility for all of the Expression profiling and proteomics work of the genomics core because of other commitments of Dr. Hinging Zhao who had been a Core Co-lead for proteomics. Dr. Zhao will still be available for proteomics consultation as initially planned. Andres Roig, M.D. and Oliver Delgado (Graduate student) have joined Projects #3 and #4 in the “TBD” positions. They are both fully trained, certified and have worked at BNL for the NSRL7 run. 
COI Name (Institution): Abrams, John  ( The University of Texas Southwestern Medical Center )
Burma, Sandeep   ( The University of Texas Southwestern Medical Center )
Caetano, Raul   ( The University of Texas School of Public Health )
Chen, David   ( The University of Texas Southwestern Medical Center )
Gazdar, Adi   ( The University of Texas Southwestern Medical Center )
Girard, Luc  ( The University of Texas Southwestern Medical Center )
Harris, T. Robert   ( The University of Texas School of Public Health )
Richardson, James   ( The University of Texas Southwestern Medical Center )
Shay, Jerry   ( The University of Texas Southwestern Medical Center )
Story, Michael  ( The University of Texas Southwestern Medical Center )
Wright, Woody  ( The University of Texas Southwestern Medical Center )
Minna, John  ( The University of Texas Southwestern Medical Center ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR proposal focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multistep pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes will also be compared in these models to that of X-irradiation. These dose risk assessments will measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. 3D organotypic culture and animal models will also allow measurement and risk estimation of “bystander” effects. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORS and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have preliminary studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we have developed plans for novel transgenic mouse models to monitor HZE particle induction of lung cancer in the whole animal in real time. The NSCOR has 4 Projects: 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation.; 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells; 3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and 4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models. These projects are supported by 4 Cores: Administrative; Cell Culture; Expression Profiling and Proteomics; and Biostatistics and Bioinformatics. In addition, with funding from the Department of Energy (DOE) contributing to this NSCOR, studies of low-LET and low dose rate radiation in the same model systems are also being undertaken. The HZE particle irradiation will be done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, and holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer and a Center for the NCI Early Detection Research Network (EDRN) program, there will be great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits: Lung cancer is the leading cause of cancer death in both women and men in the USA and in the Western world. While lung cancer is caused by smoking exposure in 85% of people, because of the large numbers the remaining 15% of cases arising in life time never smokers (~25,000 patients per year) is a sizable health problem in the USA. Likewise, 50% of all new lung cancer cases occur in former smokers (quitting over 5 years before). In all of these scenarios a major underlying question has been the role of other environmental carcinogens especially environmental radiation. One source of this is radon gas in the environment including radon gas exposure in homes with alpha particles (which represent high LET radiation). Thus, the interaction of radiation in the environment on Earth including from alpha particles in the genesis of lung cancer and quantitation of the effect of such radiation on lung epithelial cells is of major health importance. Radiation is a known carcinogenic influence but the molecular events in lung epithelial cells in response to radiation need to be determined. A key element of this is the impact of radiation on lung epithelial cells with preneoplastic lesions. Our NSCOR uses innovative technology of human bronchial epithelial cells (HBECs) developed by us (coming from 36 individuals) which have also been engineered to have known oncogenic preneoplastic changes (such as oncogenic KRAS and knockdown of p53). Our studies are providing quantitative data following high LET irradiation on these HBECs. The primary benefits from this work will be the development of quantitative risk assessment models for astronauts of high LET radiation in space. However, the results will also have major impact and benefit to life on Earth includes new knowledge of the effects on human lung epithelial cells of irradiation in terms of quantitative genetic and epigenetic changes and gene expression changes following irradiation. As part of this will be the quantitative response of DNA repair pathways to radiation including specific DNA repair components such as DNA repair enzymes and signaling pathways in lung epithelial cells. We are also determining quantitative changes in biologic phenotypes of lung epithelial cells in response to radiation which include colony formation in liquid and soft agar (anchorage independent growth) and differentiation and invasion in 3 dimensional cultures, and finally growth in vivo as either xenografts (for human cells) or endogenous lung cancers (for mouse models of lung cancer). From all of the above information we will identify novel molecular biomarkers (mRNA, DNA, protein) for lung carcinogenesis. Finally, we are learning about inter individual variation of the response of HBECs to radiation with respect to the various genetic, epigenetic, mRNA and protein expression, and signaling pathway changes.

 

Task Progress: Project 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation. Michael Story, Ph.D. Lead, Adi F. Gazdar, M.D. Co-Lead, John D. Minna, M.D. Co-Lead

The overall goals of project 1 are to establish both the low LET and the HZE radio-response of human bronchial epithelial cells (HBECs) that have been genetically manipulated, or not, with a variety of oncogenic changes that are often seen during the oncogenic progression towards lung carcinogenesis. These changes include: the downregulation of p53; the upregulation of oncogenic KRASV12; and the overexpression of wild type and mutant EGFR, as examples. These cell lines represent in vitro equivalents of preneoplastic lesions and even with multiple changes, these manipulated HBECs show only part of the malignant phenotype and are not capable of forming tumors in immunodeprived mice. Characterization of these cell lines includes standard survival analysis following radiation exposure, gene expression analysis, miRNA analysis, protein expression, DNA methylation, clonogenicity in soft agar, and tumorgenicity. Our intent is to follow irradiated cell cultures up to 4 months post-irradiation in order to examine these endpoints in cells that had survived a series of low LET exposures or specific HZE particle exposures in order to determine the effect the radiation exposure has on the oncogenic process. We will also examine inter-individual responses in those endpoints found above that enhance the oncogenic process. Our initial experiments have focused on clonogenic survival after low and high LET radiation exposures. As expected, we found that Fe56 particles are, per unit dose, far more effective at cell killing than gamma-ray exposures. Furthermore, the differences in radiosensitivity seen with low LET exposures between normal cells and cells with mutant EGFR are eliminated after Fe exposure. Si particle exposure, even though at the same energy as the Fe particles produced an intermediate killing effect. This result corresponds well with the ionization intensity differences seen via foci formation in HZE irradiated cells as determined by Project 2. Experiments to determine transformation frequency as a function of radiation exposure type are ongoing (see Project 3). Thus far, no difference has been seen four months after a 1Gy Fe exposure. This dose is highly effective at cell killing and may have eliminated cells that at lower doses would have transformed. Therefore, we are reducing the dose accordingly and re-examining transformation potential. We will continue to fill in the gaps in data for low LET exposure, and now that we have Fe and Si exposed samples, we have started the analysis of the molecular and biochemical endpoints described above, in particular, gene expression analysis. The initial data on gene expression suggest that cells do respond differently to Fe particle exposure as compared to low LET exposures. This response is likely not dose dependent but may reflect an altered kinetic response to the radiation exposure.

Project 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells. David Chen, Ph.D. Project Lead, Sandeep Burma, Ph.D. Co-Lead

With the ultimate objective of estimating cancer risks to humans from HZE particles, in this project we investigate the DNA damage responses of human cells to HZE, the link between these responses and cancer predisposition being very well established. We will take advantage of recent advances in the study of molecular mechanisms of DNA damage sensing, damage signaling, and DNA repair following low LET radiation to verify the hypothesis that HZE particles generate a different and/or unique radiation response in mammalian cells as compared to X-rays. As we are specifically interested in risks related to lung cancer development upon exposure to space radiation, we use immortalized normal human bronchial epithelial cell lines (HBECs) and three types of lung fibroblasts (WI38, IMR90 and MRC5) as model systems. Although these HBEC cultures (2D cultures) will be used throughout the project, we will also use normal or premalignant monolayer cultures (2D) and organotypic cultures (3D cultures) to examine these responses in a more in vivo tissue-like setting. The proposed research should reveal fundamental differences in cellular responses between low LET X-rays and HZE exposures in humans and in turn, provide biological markers correlated to lung carcinogenesis for the development of risk assessment and radiation protection for astronauts during future space missions. The objectives of this project are to examine the complexity of DNA damage induced by HZE relative to gamma rays in 2D and 3D cultures of HBECs and to understand the pathways responsible for repair of the complex damage inflicted by these particles and the consequences of inadequate repair responses. Using 1 GeV Fe and X-rays, we have worked out the early damage-responses, damage-repair kinetics, and persistence of DNA damage in primary human skin fibroblasts (HSFs) in great detail. Both X-rays and 1 GeV Fe particles elicit similar responses at the sites of DNA breaks. However, Fe-induced damage is repaired more slowly and a large fraction of breaks are not rejoined, thereby resulting in the higher relative biologic effect (RBE) of HZE particles. Using charged particles of increasing molecular weights (O vs. Si vs. Fe) we find that the incidence of unrejoined breaks increases with increasing molecular weight presumably due to increasing complexity of the damage induced. Detailed time courses have already been carried out with a panel of HBECs irradiated with 1 GeV Fe particles. The resulting data are currently undergoing analyses. Future plans include characterization of these responses in lung fibroblasts and epithelial cells. We have already started analyses of 2D cultures of HBECs that are in different pre-malignant stages of cancer development and in organotypic 3D cultures of lung epithelial cells and fibroblasts.

Project 3. Effect of HZE Particle Irradiation on Functional Progression of Human Lung Cancer. John D. Minna, M.D. Project Lead, Jerry Shay, Ph.D. Co-Lead

This project studies the effect of 2D (tissue culture) and 3D organotypic culture on the response to radiation from high and low LET irradiation. It is important to study the effect of HZE radiation on both the epithelial and stromal (fibroblast) components. Studying the effects of HZE radiation on lung epithelial cells should help elucidate direct mechanisms promoting initial neoplastic transformation as well as progression. Studying the effects of HZE radiation on stromal tissues may uncover other factors contributing to cancer formation and progression such as the disruption of important aspects of cellular cross- talk that promote tissue homeostasis. As described in Project 1, HBECs including HBECs engineered to contain various oncogenic changes were irradiated and then Project 3 tested them for biologic behavior relevant to cancer beginning with studies of anchorage independent growth in soft agar (a characteristic of cancer cells). The HBECs before any changes do not grow in soft agar, but do to varying degrees after these changes. We measured soft agar colony formation 1,2,3, and 4 months after HZE particle irradiation compared to control un irradiated cells. At one month there was increased colony formation in HBECs modified to contain HPV oncogenic E6 and E7 proteins, but not in any of the other 8 HBECs with various oncogenic changes or at other time points. We have also tested the effect of radiation on cells in a 3D (organotypic) system: the fibroblast collagen plug. The lung fibroblast plug closely resembles the stromal pulmonary environment. We irradiated lung fibroblast cells in regular tissue culture and in a collagen plug (3D) arrangement with low and high LET (1 Gy X-ray or 1 Gy HZE Fe56) and studied the DNA damage response and kinetics of repair. After irradiation, the plugs and cellular monolayers were fixed at specific time points (time 0 (control), 2, 12, 24, and 48 hours, and 12 days) and stained using immunofluorescence microscopy for gamma-H2AX as a marker for DNA damage which appears as large discrete foci. Radiation of low and high LET led to damage foci in 90% of cells and the amount of DNA damage was comparable up to 24 hours. However, strikingly, while the low LET radiation DNA damage was resolved by 48 hours, the high LET DNA damage persisted for up to 12 days. We are now testing the 3D cultures with epithelial cells and fibroblasts combined, and we are studying the effects of age, gender, dose rate as well as the gene expression profiles.

Project 4. Effects of HZE Particles on the Development of Lung Cancer in Novel Mouse Models. Jerry Shay, Ph.D. Project Lead, James Richardson, DVM, Ph.D.

Transgenic mouse models of human lung cancer have the advantage of a similar tumor development pathway compared to humans but with a shorter developmental period. The LA1 K-ras mouse model we have used contains a latent oncogenic K-ras allele integrated in the endogenous K-ras locus. The activation of the oncogenic allele occurs spontaneously preferentially in the lungs as the animals mature. Every animal containing the latent allele develops alveolar type II lineage lung cancer by the age of four to nine months and all animals die at approximately one year from tumor burden. Our primary objective is to determine if these animals, after low and high LET exposure, initiate or progress to cancer at increased frequency. Our long-term goal is to determine if there is an increase, decrease or no change of onset of death after HZE irradiation and ultimately to use this information for modeling risks in humans. Other end points include other cancer types, DNA damage and repair (gamma-H2AX, DNA-PKc) to correlate with the 2D and 3D cell culture experiments in projects 2 and 3. In addition, we will obtain histology and conduct expression profiles of low and high LET irradiated mouse lungs to compare to the cell culture experiments in Projects 1-3. We have conducted low LET (X-ray) at UT Southwestern Medical Center and high LET (Fe56) at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). 94 animals were irradiated with 0.2 Gy x 5 (1GeV Fe56), 97 animals were irradiated with 1.0 Gy (1GeV Fe56), 110 animals irradiated with 0.2 Gy x 5 (X-ray), and 100 animals irradiated with 1Gy (X-ray). In addition, we have 98 non-irradiated control animals. To examine acute responses (3 hr post fractionated and single dose low and high LET radiation), lung tissue was removed and RNA isolated for gene expression arrays. We are bringing on line a new mouse model of lung cancer using a transgenic mouse with a Tet-inducible mutant EGFR gene that results in lung cancer. Overall these experiments allow us to study the effects of HZE and low LET radiation on the development of lung cancer in a whole animal model.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
Articles in Peer-reviewed Journals Sunaga N, Gazdar AF, Shay JW, Minna JD. "Multiple oncogenic changes (K-rasV12, p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells. " Cancer Res. 2006 Feb 15;66(4):2116-28. PMID: 16489012 , Feb-2006
Articles in Peer-reviewed Journals Vaughan MB, Ramirez RD, Wright WE, Minna JD, Shay JW. "A three-dimensional model of differentiation of immortalized human bronchial epithelial cells. " Differentiation. 2006 Apr;74(4):141-8. PMID: 16683984 , Apr-2006
 
Fiscal Year: FY 2005  Task Last Updated:  08/22/2005 
PI Name: Minna, John D. 
Project Title: NSCOR: Lung Cancer Pathogenesis and HZE Particle Exposure 
   
Division Name: Human Research 
Program/Discipline: HUMAN RESEARCH 
Element/Subdiscipline: Radiation health 
Joint Agency Name:  
Human Research Program Elements: (1) SR:Space Radiation
Human Research Program Risks:: (1) Cancer:Risk of Carcinogenesis from Space Radiation
Human Research Program Gaps: (1) Cancer01:How can experimental models of tumor development for the major tissues (lung, colon, stomach, breast, liver, and leukemias) be developed to represent the major processes in radiation carcinogenesis and extrapolated to human risk projections?
(2) Cancer03:How can models of cancer risk be applied to reduce the uncertainties in radiation quality effects from SPEs and GCR?
(3) Cancer04:How can models of cancer risk be applied to reduce the uncertainties in dose-rate dependence of risks from SPEs and GCR?
(4) Cancer05:How can models of cancer risk be applied to reduce the uncertainties in individual radiation sensitivity including genetic and epigenetic factors from SPE and GCR?
PI Email: john.minna@utsouthwestern.edu, brenda.zielke@utsouthwestern.edu  Fax:   
PI Organization Type: UNIVERSITY  Phone: 214 648-4900  
Organization Name: The University of Texas Southwestern Medical Center  
PI Address 1: Hamon Center for Therapeutic Oncology Research 
PI Address 2: 5323 Harry Hines Boulevard, Suite 206, MC8593 
PI Web Page:  
City: Dallas  State: TX 
Zip Code: 75390-8593  Congressional District:  30 
Comments:  
Project Type: GROUND  Solicitation:  2004 NSCOR Space Radiation NNH04ZUU002N 
Start Date: 05/31/2005  End Date:  05/31/2010 
No. of Post Docs: No. of PhD Degrees:   
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No. of Bachelor's Candidates: Monitoring Center:  NASA JSC 
Contact Monitor:   Contact Phone:   
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Key Personnel Changes/Previous PI:  
COI Name (Institution): Abrams, John  ( The University of Texas Southwestern Medical Center at Dallas )
Burma, Sandeep   ( Lawrence Berkeley National Laboratory  )
Caetano, Raul   ( University of Texas Southwestern Medical Center at Dallas  )
Chen, David   ( University of Texas Southwestern Medical Center at Dallas  )
Gazdar, Adi   ( U.T.Southwestern Medical Center at Dallas  )
GIRARD, LUC  ( University of Texas Southwestern Medical Center at Dallas  )
Harris, T. Robert   ( University of Texas Southwestern Medical Center at Dallas  )
Moolgavkar, Suresh  ( Fred Hutchinson Cancer Research Center  )
Pollack,  Jonathan   ( Stanford University  )
Ramirez, Ruben   ( UT Southwestern Medical Center at Dallas  )
Richardson, James   ( University of Texas Southwestern Medical Center at Dallas  )
Shay, Jerry   ( UT Southwestern Medical Center  )
Story, Michael  ( UT Southwestern Medical Center at Dallas  )
Wright, Woody  ( University of Texas Southwestern Medical Center at Dallas  )
Zhao, Yingming   ( University of Texas Southwestern Medical Center at Dallas  ) 
Grant/Contract No.: NNJ05HD36G 
Performance Goal No.:  
Performance Goal Text:

 

Task Description: The University of Texas Southwestern Medical Center (UTSW) NSCOR proposal focuses on the development of quantitative dose risk estimates following HZE particle irradiation for the development of key genetic, epigenetic, gene expression, and cellular functional changes in the multistep pathogenesis of lung cancer in both new human bronchial epithelial cell (HBEC) and transgenic mouse models of lung cancer. The risk of developing these changes will also be compared in these models to that of X-irradiation. These dose risk assessments will measure events of both radiation-induced promotion (modification of proliferation kinetics of already-initiated cells) as well as radiation-induced initiation (mutational) events. 3D organotypic culture and animal models will also allow measurement and risk estimation of “bystander” effects. These individual risk estimates can then be combined into a model (such as a two-stage clonal expansion model) for overall risk of developing lung cancer from exposure to galactic cosmic radiation. To achieve these goals, UTSW has assembled a team of scientists who are leaders in the study and translation application of the molecular pathogenesis of lung cancer (Minna, Gazdar, Shay); in radiobiology and DNA repair (Chen, Story); a panel of expert Internal and External Advisors (from other NSCORS and lung cancer research experts); as well as expert consultants and collaborators. This team has developed a novel immortalized HBEC system that can be genetically manipulated, studied in 2D monolayer and 3D organotypic cultures. Using these HBECs we have preliminary studies with HZE particle irradiation showing that further progression toward lung cancer can be detected and quantified, that specific expression profiles for HZE irradiation exist, and that HBECs genetically manipulated to progress part of the way toward malignancy are even more sensitive to HZE-induced functional changes. In addition, we have developed plans for novel transgenic mouse models to monitor HZE particle induction of lung cancer in the whole animal in real time. The NSCOR has 4 Projects: 1. Genetic and epigenetic changes in human bronchial epithelial cells following exposure to HZE particle irradiation.; 2. Effect of HZE particles on DNA damage-sensing and repair pathways in human lung epithelial and fibroblast cells; 3. Effect of HZE particle irradiation on functional progression of human lung cancer at the cellular and organotypic level; and 4. Effects of HZE particles on the development of lung cancer in vivo in novel mouse models. These projects are supported by 4 Cores: Administrative; Cell Culture; Expression Profiling and Proteomics; and Biostatistics and Bioinformatics. The HZE particle irradiation will be done at Brookhaven National Lab, and the biostatistical analysis shows the experiments are powered to help achieve NASA mandated risk estimate confidence levels. Finally, since this group has substantial UTSW institutional commitments, and holds a NCI Special Program of Research Excellence (SPORE) in Lung Cancer and a Center for the NCI Early Detection Research Network (EDRN) program, there will be great synergism and additional resources available for the successful completion of this proposal.

 

Research Impact/Earth Benefits:

 

Task Progress: Please note that this is a new grant in FY 2005. There is no research progress to report for this period. For more information on this grant or this PI, please contact the Task Book Help Desk at taskbook@nasaprs.com or via phone at 202.479.9030 x241.

 

Bibliography Type: Description: (Last Updated: 03/15/2010) Show Cumulative Bibliography Listing
 
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