Committee - Translational Research Program Translational Research Program COMMITTEE (January, 2008.) Adam Dicker, M.D., Ph.D., Chair BREAST/TRP Dr. Woodward introduced herself as the TRP breast liaison and welcomed everyone. The stem cell symposium summary regarding future breast TRP protocols was successful however there was significant overlap with breast cancer and translational research. CTC proposals are underway and consideration was given to the need or feasibility of collecting and storing fresh samples. A feasibility study will be needed to address issues of semantics. There are no clear markers ready for prime time in multi-institutional setting. We have highlighted the importance of collecting tissue before and after treatment for future studies when markers become available. Dr. Woodward gave an update on RTOG 9804. Currently this study has collected specimens from thirty-five (35) cases. Currently there are no plans for translational research and the group has solicited ideas. In progress are plans using CTC components for RTOG 0715 a phase II study of repeat breast preserving surgery using 3D CRT and PBI. An effort is underway to make collection of CTC's a primary endpoint on this study. Dr. Steve Churma gave a slide presentation outlining the study design and progress for definitive radiation for oligomets with a purely translational endpoint. Future TRP Plans RTOG 0713 a phase III study of intensity modulated radiation therapy (IMRT) for WBI comparing hypofractionation and incorporated boost versus conventional fractionation for stage 0, I or II breast cancer. There will be collection of SNP analysis and a suggestion was made to consider biomarkers for cardiac toxicity. There will be a presentation on this at the next meeting to consider status of current data, what to collect in what setting. A randomized phase II trial of pre and postoperative conformal radiation therapy following neoadjuvant chemotherapy in patients with stage three disease paired biopsy collection planned to evaluate for markers of radiation response. Tumor Banking The RTOG Tumor Bank will be moving to University of California, San Francisco as of February 15, 2008. Dr. Frederic Waldman introduced himself as he is the new Director of the Tissue Bank and will overseeing the move and the project. Kim Dang is the new administrator for this effort. Work is in progress to put the archive inventory on-line. GASTROINTESTINAL/TRP The Gastrointestinal Translational Research Program met in San Diego at the winter 2008 semi-annual meeting of the RTOG. The agenda for this meeting included discussion of seed grants, on-going and completed pilot projects, interface with clinical gastrointestinal studies at RTOG and with other groups (R01s, P01s, SPOREs, and cooperative groups) and preparations for competitive grant renewal. The session started with Adam Dicker (Thomas Jefferson University Hospital, Philadelphia) speaking about RTOG TRP seed grants primarily focusing on availability of tissue and/or funds. He also addressed issues related to collaboration with SPORE and P01 grants at other centers, collaborations with industry, uniform processing of specimens for immunohistochemistry (IHC) and the need to turn around seed grants into publications and peer-reviewed external grants. Chandan Guha (Montefiore Medical Center, New York) spoke about logistical issues related to processing of RTOG specimens. With adoption of the HistoRx as the preferred platform for all future IHC studies, Adam outlined the experience from Calgary where the system has been standardized, validated and optimized for high-throughput analyses of stained tissue microarrays. Anthony Magliocco (Southern Alberta Cancer Research Institute, Calgary) further emphasized the excellent interobserver reproducibility, ability to obtain quantifiable data with segmentation into cytoplasmic vs. nuclear, epithelial vs. mesenchymal, etc, and the ability to multiplex multiple antibodies simultaneously. The scientific session began with a presentation by James Farrell (University of California, Los Angeles) outlining the results of their investigations in pancreatic cancer using RTOG 9704 specimens. Using tumor block DNA extractions for single nucleotide polymorphism (SNP) analysis, Farrell's group identified human equilibrative nucleoside transporter (hENT1) as predictive for improved overall survival and disease-free survival among patients treated adjuvantly with gemcitabine but not among patients treated with 5FU. This is in keeping with the knowledge that hENT1 modulates gemcitabine transport into cells but is unrelated to 5FU uptake or metabolism. Furthermore, these findings were corroborated by similar data on IHC analysis. These results represent the first identification of a predictive marker for gemcitabine response. This group also evaluated SNP changes in ribonucleotide reductase subunit protein M2 (RRM2), a rate-limiting enzyme in DNA synthesis and repair. The TT allele was found to predict for worse overall survival among pancreatic cancer patients treated adjuvantly with gemcitabine but not among patients treated with 5FU. Lastly, a preliminary analysis of immunohistochemical staining for CD68, a marker for tumor-associated macrophages could potentially help tumor cells with invasion and metastasis, demonstrated higher levels of CD68 were associated with lower overall survival among all patients treated adjuvantly with chemoradiation therapy (HR=1.63, p=0.02). Additional mechanistic and correlative investigations are underway. In addition, the UCLA group has systematically re-categorized toxicity data from RTOG 9704 to include temporal profiles of toxicity scores during the course of adjuvant treatment so as to better evaluate the correlation between the SNP analysis and toxicity outcomes. These studies, while not directly involved with the UCLA basic science P01, could dovetail with the overarching theme of that P01, potentially permitting some crosstalk between the RTOG Gastrointestinal Translational Research Program and the P01. There was a discussion about interfacing with the MD Anderson pancreas SPORE where Donghui Li (MD Anderson, Houston) had submitted and received funding for a seed grant to look at a battery of SNPs as potential predictors of outcome in adjuvant treatment pancreatic cancer. This would corroborate, in a different subset of patients, the MD Anderson group's findings among their patients. Unfortunately, there is minimal remaining tissue and/or DNA from the UCLA analyses but efforts are underway to share the raw data from SNP analyses so Donghui can run her analyses using these data. As a second means of achieving these results, Deborah Citrin's group is also working on amplifying DNA from tissue specimens - this DNA could also be used for independent validation of the MD Anderson findings. Chandan presented broad brushstrokes of the specific aims of the Gastrointestinal Translational Research Program for the future. These include (i) identification of prognostic and predictive molecular, genetic and epigenetic markers would serve as guides to therapy and predict normal tissue toxicity in gastrointestinal malignancies; (ii) identification of underlying molecular, genetic, and epigenetic mechanisms of therapeutic resistance to chemoradiation therapy in gastrointestinal malignancies and devising strategies to overcome resistance to therapies; and (iii) identification of immune evasive mechanisms in gastrointestinal malignancies and designing therapeutic strategies of immunomodulation of chemoradiation therapy in gastrointestinal malignancies. Within the Gastrointestinal Translational Research Program, the biomarker identification efforts will be organized with separate themes/groups that include DNA repair, growth factor signaling, angiogenesis, hypoxia, histone modification/promoter methylation, inflammation/cytokines, immunomodulation, stroma/epithelial-to-mesenchymal transition (EMT), and stem cells. In keeping with these themes, two groups presented their laboratory findings in support of their application for seed grants. Qinghong Zhang (Oregon Health Sciences University, Portland) presented data summarizing her group's identification of CtBP as a master regulator of EMT. Investigations in her laboratory have confirmed CtBP functions as a transcriptional corepressor of epithelial and pro-apoptotic genes linked to EMT. In a panel of human head and neck, colon, breast and renal cancer specimens, CtBP was uniformly overexpressed - similar data in pancreatic cancer cell lines and human pancreatic cancer specimens is currently lacking. These and other studies documenting its role in pancreatic cancer invasion, metastasis and response to radiation therapy form the basis of this seed grant application. Howard Safran (Brown University, Providence) spoke on behalf of the Gastrointestinal Cancer Steering Committee and outlined the direction being taken by the clinical committee. He underscored the need for the clinical and translational groups to work jointly on current and future gastrointestinal studies conducted by the RTOG. Tapas Hazra (University of Texas Medical Branch, Galveston) and Sushovan Guha (MD Anderson, Houston) presented data outlining the role of NEIL2 in transcription-coupled repair and repair of oxidized DNA bases. Interestingly, a search of the NCBI database of EST counts identified no NEIL2 in any of the pancreatic cancers but abundant NEIL2 in normal pancreas. Paul Okunieff (University of Rochester Cancer Center, Rochester, NY) suggested they investigate the role of NEIL2 in repairing DNA damage caused by oxidized bases formed during radiotherapy. Adam Dicker suggested they also evaluate circulating tumor cells. These presentations concluded the scientific session of the Gastrointestinal Translational Research Program. Subsequently, a discussion of other areas of interest to the attendees ensued. Eileen O'Riley (Memorial Sloan Kettering Cancer Center, New York) inquired about routine analysis of biopsy and/or surgical specimens for Kras mutations and epiregulin/neuregulin overexpression among patients treated with EGFR inhibitors. These studies are being incorporated as correlative components of future gastrointestinal study group trials. Adam Berger (Thomas Jefferson University Hospital, Philadelphia) enquired about the committee's response to a seed grant jointly submitted by him and Dan Laheru (Johns Hopkins University, Baltimore) to evaluate the role of DCK as a marker of gemcitabine sensitivity. The committee will look into this and get back to him. Lastly, given the increasing interest in Ang1 and Ang2 inhibitors as potential radiosensitizing agents, Chandan asked the Amgen representative if he would be willing to work with individual investigators to evaluate their role as potential radiosensitizing agents. These discussions with Amgen would continue offline. GENTIOURINARY/TRP The following items were discussed. RTOG 0612: This protocol involves the acquisition of frozen and formalin fixed tissue for comparison and microarray analysis. The protocol is linked to RTOG 0232. Because of RTOG 0612 is being linked to RTOG 0232, there has been slow accrual. Participants were encouraged to try to accrue to RTOG 0612. We also discussed an additional protocol being developed by Dr. Chakravarti RTOG 0827, which involves a similar objective, but using tissue taken from prostate biopsies acquired when patients undergo fiducial marker implantation prior to external beam radiotherapy. This protocol is in development. Action items: Improve enrollment into RTOG 0612 Complete development of RTOG 0827 There was a considerable discussion about the accuracy and reproducibility of ChromaVision versus HistoRx. The HistoRx platform is felt to be more reproducible between laboratories. Dr. Chakravarti was concerned about the results with pAKT using Chromavision, which appeared to be counterintuitive. The HistoRx results were the opposite and were more in line with expectations. There was general agreement we should stain markers in the tissue of microarray available from RTOG 8610 and compare these results with the prior ChromaVision results. This will give us an idea of the concordance between these different methods before we move on to stain other markers in RTOG 9202 and beyond. We have been comparing manual counts with ChromaVision counts, but such comparisons are difficult, particularly when using parameters involving intensity of staining. The HistoRx confirmation will point us in the right direction for future studies. The group in Edmonton have been willing to collaborate with us on the analysis of markers by HistoRx. Action Items: Stain for Ki-67 MDM2, p53, Bcl-2 Bax, p16, Cox-2 and PKA RTOG 8610 for HistoRx analysis. Review of markers from RTOG 8610 & 9202: Progress on the analyses and publication of markers quantified by immunohistochemistry using the ChromaVision image analysis system was reviewed. The group continues to be productive in terms of publications with a new paper in Lancet Oncology over the last six months, in addition to other papers. Paul Nguyen presented an overview on p27, data on which are mostly from radical prostatectomy studies. He requested that we run p27 in prostate tumor samples from RTOG 9202. The group was in agreement this was worth doing. He will submit a pilot grant to the RTOG. There was discussion about fusion genes, such as TMPRSS: ETS. There has been enthusiasm for measurements of this gene in the past and the group was enthusiastic about pursuing this further. We will pursue talking to Dr. Arul Chinnaiyan again to see if he has an interest. Dr. Arnab Chakravarti remains interested in running Survivin, pAKT, pMAPK, and PTEN in RTOG 9202. Funding would be needed to perform these studies. The analyses described will be prioritized by funding. Action Item: Develop model of biomarkers from RTOG 9202. Upgrade on RTOG 9408 Markers: Dr. Mark Ritter is the PI on a grant related to these analyses. Ki-67 and p53 should be completed very shortly. Staining for Cox-2 and MDM2 are planned in the near future. Since these markers have already been established using ChromaVision, the work will continue to use ChromaVision. Dr. Ritter has accessibility to a HistoRx machine, and may be able to validate a subset of these markers on the HistoRx machine. We anticipate over the next six months permission will be granted by the NCI for changing RTOG 9408 to a biomarker endpoint, so the data can be revealed. The trial was started 12 years ago. Action Items: Complete staining and analysis of all four biomarkers over the next six months. Plans for RTOG 9413: An R21 (PI: A. Pollack) is under consideration for the analysis of samples in RTOG 9413 have been shown to be promising in RTOG 9202. Also mRNA analysis is planned. These studies will be pending the results of the HistoRx studies. Probably, no new studies will be initiated over the next six months unless there is additional funding from the R21. Plans for RTOG 9910: It appears RTOG 9910 still has a long way to go before endpoints are reached and analyses may be undertaken. We will continue to be updated on progress related to reaching the primary endpoint and when it looks like it is in reach, we will pursue biomarker studies. Dr. Pollack has applied for funding for the analysis of E2F-1 and MAD2 in RTOG 9202 (R01 application will be resubmitted) as well. Update on bladder protocol markers: An RTOG pilot grant has been awarded to Bill Shipley for the analysis of VEGF. These analyses are pending HistoRx studies. Other items: There was discussion of the collection of blood products for SNP analysis. We will check the protocols in which SNP analysis has been added. In particular, we will check if RTOG 0126 has added this. GYNECOLOGY/TRP We were unable to find out the exact number of samples collected on RTOG 0417 and RTOG 0418 because the site has not been updated, but we encouraged everyone to submit tissue. RTOG 0128 microarray and tissue microarray update: No one from MDACC was present to discuss their plans for using RTOG 0128 data to compare the results with their microarray data in cervix. However, they are planning to compare the microarray data to theirs, as there was not enough tissue to run new microarrays. Dr. Doll and Magliocco from Calgary showed their preliminary data using the HistoRx machine with the RTOG 0128 tissue microarray. They discussed some of the potential pros and cons. They have preliminary data ready for analysis at Headquarters. They inquired about the possibility of getting an H and E slides of this TMA, asked about the "mid" sample, and asked about possibly constructing a TMA of just the pre and post samples. Dr. Weidhaas presented the results from Yale in analyzing the RTOG 0128 microarray data with clinical outcomes. They were able to find a gene expression change pattern of 14 genes that predicted local control or failure. This is statistically significant and in preparation for publication. Dr. Doll introduced the group to the CARO group, a collection of cervical cancer samples where hypoxia markers are being studied in Canada. They have requested (through the seed grant mechanism) to compare the results on RTOG 0128 tissues. Attendees asked some questions about the findings on the microarray data and stem cells, which were discussed. HEAD&NECK/TRP Summary of Dr. Gillison's Presentation Dr. Gillison reviewed the evidence of the association between HPV and head and neck squamous cell carcinoma, especially oropharyngeal primary tumors (OPSCC). The clinical differences of HPV (+) compared to HPV (-) tumors were younger age, high risk sexual behavior, better survival and increasing incidence since early 90's. Clinical outcomes in 7 independent studies were consistent in improved survival in HPV (+) patients. In ECOG 2399, 63% of OPSCC was HPV (+) and had 2-year overall survival of 95% when HPV (-) patients were 62% (p=0.005 Log-rank test). Also, several HPV detection methods were compared and combination of p16 IHC and HPV16 in situ hybridization provided sensitivity 95% and specificity 98% at the Johns Hopkins University Laboratory. Discussion Regarding the Sample Size of RTOG 0522 There is a concern RTOG 0522 may be underpowered because the sample size was calculated with a historical control when HPV incidence has been increasing. The control group may have better outcome than expected and the small gain in the experimental group may not be detected with the current sample size. Currently about 70% of the patients in both arms are OPSCC. About 40% of tumors from RTOG 0522 are in the RTOG tissue bank. Action Item Christine Chung will visit the RTOG tissue bank once it is moved from Utah to San Francisco to assess the quality and quantity of the tumor samples from RTOG 0522. Appropriate tumors will be made into a tissue microarray. For tumors with limited amount of tissue, the HPV testing has to be prioritized by the H&N core committee because there are other approved correlatives studies embedded in the protocol. The H&N core committee will decide whether to test for HPV retrospectively or prospectively. In the mean time, Tom Pajak and Maura Gillison will work together to re-calculate the sample size of RTOG 0522 based on the data from E2399. The importance of tissue collection under RTOG 0514 was reiterated. The update on tissue inventory and IHC Quantification Optical Imaging Standards will be discussed at the next meeting. LUNG/TRP 1. The first 20 minutes was a presentation of data on FISH analysis of RTOG 0324 by Dr. Hak Choy. RTOG 0324: A Phase II Study of Cetuximab (C225) In Combination with Chemoradiation in Patients with Stage IIIA/B Non-Small Cell Lung Cancer (NSCLC). This study has enrolled 93 patients, 87 of whom were evaluable for analyses. Though not mandatory, tumor specimens were collected from 51 (59%) of the evaluable patients for correlative studies. TRP: EGFR expression: Both IHC and FISH are completed. (ASTRO abstract published for IHC, 2006; ASCO abstract for FISH submitted, 2007) 2. The second 20 minutes: Dr. Yuhchyau Chen: Lung TRP Update, Publications, Grant Renewal, Other Business: 2.1 Lung TRP Mission: Identify molecular, genetic and epigenetic signatures for lung cancer prognosis and invasiveness. Based on marker information, prospectively incorporate molecular, genetic and epigenetic markers to lung clinical protocols and design therapeutic interventions for molecular pathways, thus reducing individual risks of cancer recurrence. Identify molecular, genetic and epigenetic signatures for radiation normal tissue injury after thoracic irradiation. Based on normal tissue response marker information, prospectively incorporating molecular, genetic and epigenetic markers to lung clinical protocols to minimize individual risks of radiation side effects. 2.2 RTOG 0617 TRP Tumor markers - ERCC-1, XPA, XPC, XPD, XPF, XPG, SF-2, SNP related to DNA repair gene; Apoptosis related RNA expression (TRAIL, caspase, BCL-2); and hypoxia marker (osteopontin), P53, Ras. 2.3 Markers for Normal Tissue Injuries: Circulating markers - VEGF, interleukins, TGFß and others. New markers: Tumor stem cells, and circulating endothelial cells -marker for angiogenesis (RTOG stem cell symposium, 01-18-08), MN-RET. Tumor markers - ERCC-1, XPA, XPC, XPD, XPF, XPG, SF-2, SNP related to DNA repair gene; Apoptosis related RNA expression (TRAIL, caspase, BCL-2); and hypoxia marker (osteopontin), P53, Ras. Circulating markers - VEGF, interleukins, TGFß and others. New markers: Tumor stem cells, and circulating endothelial cells -marker for angiogenesis (RTOG stem cell symposium, 01-18-08), MN-RET. 2.4 TRP recent progress: Major improvement in tissue procurement in recently completed phase II trial (RTOG 0324) Successfully analyzed the tumor specimens for EGFR pathway by the leading experts (RTOG 0324, IHC [Dr. Kian Ang] and FISH [Dr. Fred Hirsch]. Seed grant: success in yielding data, publications, and external grants. Academic leadership: Reflected by externally funded grants on lung translational research by lung TRP members. 3. Dr. Corey Langer: Medical Oncology gave a 15 minute talk on molecular targeted therapy in lung cancer. 4. Dr. Jeff Bradley/Dr. Mitchell Machtay gave a 15 minute talk about the PET and lung study. 5. Dr. Spring Kong: Update on blood sample processing and its significance to lung TRP for 15 minutes. 6. The meeting was opened to questions. Translational Research Program COMMITTEE (June, 2007) Adam Dicker, M.D., Ph.D., Chair BREAST/TRP Funding: Adam Dicker highlighted the goals and expectations of the group - ideally an example of productivity by the January meeting. He also explained the role and availability of TRP seed grants, $50,000 per year to support the mission of the RTOG. He also discussed the role for better communication with the advocacy groups to engender support for these types of end points. Tissues: Liz Hammond reported on current material in RTOG for breast cancer. The DCIS material is held by the CALGB. Liz highlighted the move towards intergroup collaboration regarding tissue collection and noted we can get access to this material as well as material from trials run by other groups. Nour Sneige has slides from a proportion of the DCIS cases to validate an on-line teaching tool for DCIS pathology. In spite of the plans written in the protocol in RTOG 0413 for specimen collection, only one case has been submitted on this protocol. Technology: Wendy Woodward reviewed the technical issues involved in collecting CTCS. Veridex is the only FDA approved approach for this. RosetteSep and Easy Sep are available and less expensive but also less quantitative and require more starting material. RNA can be obtained from both approaches however the quality appears better in EDTA in the veridex cell save preservative. Time to analysis is critical - overnight shipping will be required. Samples should be timed and dated to be included in the analysis. We will plan to put together a CTC kit that can be incorporated into trials. Dr. Woodward will work with Liz Hammond on this. GASTROINTESTINAL/TRP Approximately twenty five attendees were present for the GI TRP meeting. Dr. James Farrell gave a presentation on correlative studies with tissue arrays from RTOG 9704. Two manuscripts on the role of HENT1 and RRM2 in predicting outcomes in RTOG 9704 patients are currently undergoing internal RTOG review. We should be able to submit them once approved by the RTOG publications committee. The statistical office at headquarters is to be involved in the final data analysis and kept abreast of the results. Steps to facilitate current and future RTOG 9704 correlative analysis, including possible release of limited RTOG datasets to us at UCLA to do our own analysis were discussed. The use of tissue microarrays from RTOG 9704 was discussed. Discussion ensued about maximizing the benefits of the tissue resources of RTOG 9704. Immunohistochemistry core laboratory for RTOG. Digital scoring better than manual scoring with better more reproducible results. Dr. Adam Dicker suggested collection of Buffy coat for single nucleotide polymorphism (SNP) analysis in the future. Members were encouraged to fully participate in the committee. Grant ideas were encouraged. Dr. Adam Dicker specifically encouraged submission of seed grants. GYNECOLOGIC/TRP Sample collection on RTOG 0417 was discussed. To date, there have been a high percentage of cases submitting fresh tissue. The optimal material for proteomics was discussed. It was decided in the future, plasma would be collected as well as sera. Dr. Adam Dicker also suggested collection of Buffy coat for single nucleotide polymorphism (SNP) analysis in the future. Consequently, the next amendment will include a collection for plasma and blood for future SNP analysis. Next, sample collection on RTOG 0418 was discussed. Investigators were encouraged to have their research associates submit blocks. The group was encouraged to consider study ideas for this trial. Dr. Joann Weidhaas gave an update on gene expression analysis from cervical cancer on trial RTOG 0128. The micro array data analysis is proceeding at Yale University. Headquarters has provided the data to investigators at Yale. The statistical office at headquarters will be involved in the final data analysis and kept abreast of the results. The use of tissue micro arrays from RTOG 0128 was discussed. Dr. Corrine Doll and Dr. Tony Magliocco have submitted an RTOG seed grant to evaluate differential protein expression for a number of predictive and prognostic biomarkers. Dr. Adam Dicker, Vice-Chair of the Translational Research Program was supportive of this grant concept. Drs. Doll and Magliocco were encouraged to focus on the protein aspects of this grant submission and not the micro-RNA aspects. Members were encouraged to fully participate in the committee. Grant ideas were encouraged. Dr. Adam Dicker specifically encouraged submission of seed grants. HEAD & NECK/TRP RTOG 0514 - Stuart Wong raised proposal whether the protocol should/could be amended to permit study of radiation damage/repair. This would require additional tissue collection during therapy. Discussion ensued regarding if this was important, which tissue to collect, and ideal time point of collection. All agreed the scientific question is relevant and important. A suggestion was made that blood (serum, mononuclear cells), buccal scraping should be collected. Optimal time of collection should be before clinical effects of RT are set in between 7-14 days post start of RT. ACTION ITEM: Stu has discussed this with Randy. Further input from Liz Hammond will be required. Issue will then be brought back to HN TRP group. IHC Quantification Optical Imaging Standards - The group discussed RTOG TRP initiative to have a standard platform across all TRP disease sites. Adam discussed overview/rationale, and progress of contractual agreement with HistoRx. Discussion followed a well-established, validated and published methodologies have been used by members of HN TRP group. This discussion was specifically pertinent to the design of Dr. Le's TRP proposal. Recommendation was made by the group, rather than adopting platform change outright, validation and comparison of methodologies should be examined first. ACTION ITEM: Pilot validation study was recommended: Stanford TMA will be used to study a limited set of markers (ecadherin, lox, egfr, heregulin and amphi-heregulin). Comparison of standard quantification versus AQUA. OTHER TRP ISSUES - Not discussed due to time constraints. HN TRP Sub-Committee Meeting Time - The general TRP meeting conflicts with general HN Committee meeting. Need to have an HN liaison at general TRP meeting. Need to find an acceptable time for HN TRP. Formal request to have HN TPR added to agenda was placed in April 07 but was not completed. ACTION ITEM: Separate conference call is planned. HPV. HN steering meeting discussed Gillison's TPR proposal to exam HPV in RTOG 9003 specimens. Insufficient tissue was not available. Alternatives were discussed eg RTOG 9501. Suggestion was raised to exam RTOG 0129 tissue. ACTION ITEM: Will discuss on upcoming conference call. LUNG/TRP Dr. Yuhchyau Chen called meeting in session at 4:30 pm. Dr. Chen acknowledged Dr. Dicker and Mr. Tom Wudarski in guiding the steps towards setting up the Lung TRP Subcommittee. Dr. Dicker gave a brief opening remark on the RTOG TRP and lung TRP mission. Dr. Hak Choy commented on lung TRP focus: Predictive markers (not prognostic markers) for toxicity and survival in lung cancer. LUNG TRP UPDATE (Dr. Chen) a. Tissue Procurement History: 45 total (blocks plus slides, from 6 clinical trials from 1993 to 2004): Major barriers: 1) most diagnoses were made by FNA (inoperable lung cancer), limited tissue size for translational research 2) lack of awareness from clinical protocols 3) lack of commitment to obtain tissue. Progress: RTOG 0324 (ChemoRT + C225 for stage III NSCLC) has achieved unprecedented tissue procurement rate: 1+ block=32, 5+ unstained slides =33 I.e. 65 tissue samples/78 cases= 83% procurement rate Tissue procurement protocol in preparation: Dr. Carolyn Jones (Thoracic Surgery) and Dr. Chen, from the University of Rochester for resected stage I, II, and III non-small-cell lungs, small cell, thymoma, mesothelioma, and normal lung tissue. b. Lung Correlative Studies RTOG 0324 (chemoRT + C225 for stage III NSCLC) specimens have been analyzed for EGFR-1 expression by both IHC stain (Dr. Ang) and FISH (Dr. Hirsch). Dr. Komaki will be presenting the IHC study result at ASTRO. The data on FISH have been sent to RTOG biostatistics. c. Lung TRP Current Status: Build TRP in all New Clinical Protocols For all new protocols, add collection of serum, plasma, buffy coat, and urine. (Dr. Hak Choy and PI of clinical study to support) Include molecular markers, gene signatures, cytokine markers and new markers pertinent to lung "cancer" and to "normal tissue": (ERCC-1, 5 signature genes of invasiveness, IL-1, IL-6, IL-8, TGF-ß and others) Standardize protocol for blood/body fluid and buffy coat collection for lung studies: Spring Kong (seed grant) and Yuhchyau Chen PRESENTATIONS a. S0424 SWOG/Intergroup Molecular epidemiology lung study update: Dr. Chen b. Lung tissue procurement update: Dr. Hammond (deferred) c. Tumor EGFR expression: RTOG 0324 phase II chemoRT + C225 study: Dr. Komaki d. Seed grant update on plasma processing: S. Kong e. Esophageal cancer stem cell and the possibility for lung cancer studies: Dr. Chang f. ERCC 1 pharmacogenomic study concept: Dr. Massard COMMENTS and QUESTIONS Several comments were made during the meeting session. Overall this first lung TRP meeting was perceived as a success since the meeting was carried out with great enthusiasm and dynamic discussions by the participants. For future improvement, a few suggestions were made: - extending meeting time from one hour to two hours. - requesting a larger room for future meetings. - needing a TRP lung imaging component (Dr. Machtay has agree to next lung TRP meeting for this focus). - future presentation should focus on RTOG translational research, and not on basic science. We suggest presenters e-mail slides to Dr. Chen before the meeting to review ahead and make suggested changes. We may limit the number of slides to 10 per presenter. - Dr. Chen to provide a summary of TRP projected progress in the next 6 months in two slides to Dr. Dicker for the Sunday Executive Committee Meeting and the Grant Renewal Meeting. SARCOMA/TRP 1.0 Dian Wang 1.1 Update on TRP Component of RTOG 0603 1.1.1 Dr. Wang discussed this in limited detail because of time constraints. Concern was expressed about the issue of biopsies of skin and quality of life. It expressed this would be difficult. 1.1.2 Recommended a patient advocate be involved to gain support for involvement in this protocol, Mary Lou Smith from Chicago is such a person. Dr. Hammond knows this person. 1.2 My notes do not reflect the extent of the discussion and concerns expressed. The protocol has been sent to CTEP. Result is pending. 2.0 Dr. Meg von Mehren discussed RTOG 0132, A Phase II Trial of Neoadjuvant/Adjuvant STI-571 for Primary and Recurrent Operable Malignant GIST Expressing the Kit Recepter Tyrosine Kinase (CD117): Problems. 2.1 TMR Bioinformatics are at Johns Hopkins, not sure what access we have to data. Solution. It is being investigated. Dr. Godwin will be called. 2.2 Sixty three patients, 24 blocks, 17 at Fox Chase. Problem is Fox Chase Cancer Center has not sent out blocks the RTOG Tumor Bank in Salt Lake City, UT. Dr. Von Mehren will address this issue. 2.3 Mike Heinrich: Mutational analysis of fixed tissue. Material in the biorepository was 24 blocks and the rest were submitted as a bunch of unstained slides. These will not be adequate of the purposes of this study. Headquarters should help with this. 2.4 ACRIN fails to communicate to RTOG and vice - versa. This will be addressed by RTOG. This should be solvable since both ACRIN and RTOG are in the same building. 2.5 Pathology 2.6 Most of the patients are alive. Idea of SNIPS analysis was discussed. Need buffy coats. Would need to amend protocol, consent for blood drawing and follow up. There is money to do this. Problem with RNA quality. RNA latter not available for this study. 2.7 Toxicity 2.7.1 Bleeding and Fistula 2.7.2 Usually same as patients with advanced GIST 3.0 Dr. Lev very briefly presented her work. - She has received second seed grant. Strong consideration will be given to her presenting her work at the January meeting 4.0Dr. Kraybill briefly reported on RTOG Tissue protocol effort. A concept sheet for this will be presented to the research strategy committee June 24. Translational Proposals Currently Under Evaluation of In Progress: Andy Minn presented the rationale for a new concept in the Breast Cancer Working Group. Ultimately the most feasible incorporation of all goals may be to plan to do gene array analysis (using a previously described predictive signature describe by Dr. Minn) on FFPE tissues from metastatic biopsy accessible sites, and collect peripheral blood in cell save for quantitation of circulating tumor cells. The same approach will be used to collect CTCs on the PBI for re-irradiation after IBTR protocol - concept approved. Peripheral blood will be collected in a separate tube for SNPs to be correlated to toxicity. Peripheral blood will be collected for SNPS for the proposed Phase III IMRT concept. Additional material will be collected as feasible - to be considered further as the concept moves further. A new concept has been proposed but not yet discussed at the Breast Cancer Working Group due to time constraints. This is a pre-op chemo radiation (concurrent herceptin) versus post-op radiation trial as a model for translation radiation sensitizer studies. Proposed pt group are Stage III H2N+ patients who will have a mastectomy and post-mastectomy radiation based on stage. Endpoints in development. Translational Research Program COMMITTEE (February, 2007) Adam Dicker, M.D., Ph.D., Chair BRAIN/TRP Overview: The focus of this meeting was the planned PO1 grant submission. The organizational structure and timelines were reviewed in detail. Our objective is to submit this grant for the October 1, 2007 deadline. This goal was viewed as ambitious, but potentially achievable if crucial preliminary data can be obtained in time. The observation was made that the projects are morphing into one another quite well and are becoming quite interdependent. A meeting will be held at MD Anderson Cancer Center in April of 2007 with the objective of generating our first complete PO1 draft. An external scientific review committee will be established to evaluate our initial drafts. The goal here will be to receive their comments by June of 2007 to have several months to make necessary revisions to the PO1 grant. A PO1 budget will be established by June of 2007, with all institutional LOIs executed with RTOG HQ by this time. Project #1: Transcriptional/Epigenetic mechanisms of Treatment Resistance in High-Grade Gliomas. Ken presented his 38-gene predictor data, which has demonstrated great promise. Although only one gene (aquaporin) was held in common with Monika's top 38-gene set, 87% of genes in Ken's dataset were identified as survival genes of variable predictive significance within Monika's dataset. Monika discussed her findings with regards to gene expression profiling and MGMT methylation status in the TMZ-treated EORTC dataset. It appears that MGMT methylation status remains the most powerful prognostic/predictive marker. However, the gene expression analysis data adds some potentially important information. Monika has identified EGFR expression, combined with MGMT methylation status as being a potentially important predictor in TMZ-treated patients. Further, there appear to be distinct gene clusters such as the Hox, Stromal, etc. that appear to have important predictive value. Pierre Robe presented data from Harvard regarding GSEA and ACCESS analysis, which examines gene chip data through the viewpoint of larger molecular pathways rather than a gene-by-gene basis. Pierre described specific pathways that appear to have important prognostic and predictive value in high-grade glioma patients. It was felt by the group that the metagene predictor set may be important for general prognostic/predictive value, but larger pathway analysis such as GSEA and ACCESS may prove important in uncovering actual resistance mechanisms in gliomas. Greg Jones from Oncomethylome presented data on their global methylation chip. An important underlying question remains unanswered is whether MGMT methylation patterns trump global methylation patterns in terms of prognostic/predictive value. Oncomethylome has designed a chip containing 80 genes known to contribute to chemotherapy and radiation resistance to help answer this question. The sensitivity and specificity of these chips have been reported to mirror single-gene methylation-specific PCR data. Oncomethylome is very eager to collaborate with RTOG on investigating their new chip in gliomas. Mike Kuo presented data on radiographic correlates of gene expression profiling patterns, which was viewed as promising, but a bit preliminary at this stage. Action Items: 1) Validation of Ken's 38-gene signature data on TMZ-treated patients. Pierre and Arnab will provide Ken with additional tissues (with pertinent clinical information) to help address this issue and lead to the development of the appropriate Illumina platform for RTOG 0525-treated patients. 2) The newly-acquired 38-gene signature data derived from TMZ-treated patients will be cross- correlated with Monika's EORTC dataset as well. 3) Based on the newly-acquired (Ken's, Pierre's, and Arnab's) and EORTC TMZ-treated patient datasets, respectively, it would be ideal to identify larger molecular pathways associated with treatment resistance in malignant gliomas using GSEA and ACCESS-based analysis. This would be complementary to the predictive/prognostic metagene dataset in that more mechanistic insights (potentially) can be obtained from such a pathway-based analytic approach. 4) Ken, Arnab, and Pierre will provide tissues to Oncomethyome to test their global methylation chip. If promising compared to single gene methylation controls, this will serve as preliminary data/justification for incorporating such global methylation analysis into the PO1 grant. 5) Using Jann's in vivo model, it would be ideal to generate a metagene/pathway signature, inducable by RT+TMZ, that is predictive of response and survival outcome. Indeed, this may also provide important mechanistic data that cannot be obtained through pretreatment profiling alone. 6) Mike Kuo will work with Monika and Ken to sharpen and define hypotheses related to imaging correlates. Project #2: DNA Repair/Chromatin Remodeling Resistance Pathways: PARP-1 data from the Chakravarti/Plummer/Calvert Labs were presented in detail. Our group has a collection of PARP- 1 inhibitors from Abbott, MGI, AstraZeneca, and Inotek. Our study based on the Inotek compound, PJ-34, has been obtained and will be soon submitted for publication. PARP-1 is known to have dual functions in human tumors: DNA repair (pro-survival) and suppression of Metabolism (pro-death). Therefore, the biological significance of PARP-1 inhibition depends highly on which PARP-1 function is paramount within a given tumor. We examined the efficacy of PARP-1 inhibition in the setting of RT+TMZ and TMZ alone. Our findings strongly suggest in tumors where PARP-1's primary function is to suppress activation of AKT and of downstream metabolic pathways, PARP-1 inhibitors hold the potential of increasing resistance to radiation. Where PARP-1's primary function is to enhance ds-DNA repair, PARP-1 inhibitors hold the promise of enhancing the sensitivity of tumor cells to radiation. PARP-1 inhibitors appear to have somewhat more consistent effects in the presence of TMZ alone, with overall efficacy dependent on baseline activation of PI3K/AKT signaling. Fen Xia presented data on the possible role of BRCA1 in DSB repair in gliomas. Her findings are preliminary, but thought will be given in incorporating her research into the PO1, depending on the nature and maturity of her findings at the time of submission. Hilary and Ruth are also actively investigating DNA-PK and ATM as possible mediators of resistance in high-grade gliomas. Unfortunately, they were unable to make the meeting to present their findings, but will update us on their findings during the next teleconference. Hilary and Ruth have already written a draft of their hypothesis and specific aims, which build upon our initial findings, with regards to PARP-1, MGMT, DNA-PK, and ATM. The prognostic and predictive values of critical DNA repair molecules will be evaluated in this project as well using tissues from the tissue bank for RTOG 0525, 9813, and the nondeleted AO trial. Action Items: 1) The Abbott, MGI, AstraZeneca, and Inotek PARP-1 inhibitors will be tested head-to-head in the setting of RT+TMZ and TMZ alone to identify the most effective agent(s). Markers of resistance such as NAD and PI3K-related molecules will be examined. 2) Hilary and Ruth will continue to investigate other potential markers of resistance along the DNA repair pathways such as ATM and DNA-PK. 3) Fen will investigate the role of BRCA-1. 4) The collaboration with oncomethylome, as described above in Project #1, will not only serve to confirm the the predictive value of MGMT, but will also identify whether methylation of other DNA repair genes has significant predictive/prognostic value in gliomas. 5) The role of these DNA repair pathways in stem cell-mediated resistance will be examined. 6) We have already reported important tie-ins to critical signal transduction pathways, which will serve as a solid foundation upon which to be built upon in the coming months. Angiogenesis: Phyllis Wachsberger described her hypothesis and specific aims, which focus on signal transduction pathways regulating the pro-angiogenic phenotype. Her aims focus on identifying critical connections between the EGFR-PI3K-HIF-1 pathways and the angiogenic phenotype in gliomas. Phyllis has identified signal transduction pathways that appear to be critical in regulating resistance to RT+TMZ in isogenic U87 cells in vitro. Her findings suggest that tumors with EGFR OE + PTENwt are the most sensitive to RT, whereas EGFRnull+PTENwt are the most resistant cells to RT. Ken's, Monika's and Arnab's clinical data suggest that activated AKT and/or EGFR OE are associated with increased treatment resistance. Therefore, there was some concern raised about whether the in vitro isogenic U87 data is reflective of clinical reality. Indeed, isogenic models may prove more useful in studying specific interactions between molecules within a signal transduction pathway, rather than to assay the efficacy of a given therapy. It was suggested to Phyllis the associations between signal transduction pathways and treatment resistance, especially where questions of sensitivity to anti-angiogenic agents are at issue, may be better studied in vivo versus in vitro. Indeed, Jann's xenograft model and Howard's stem cell (which can be grown intracranially as well) models may be preferable to answer such questions. Phyllis also outlined her strategies to determine the most effective anti-angiogenic therapies in malignant gliomas (e.g. VEGF-trap, AZD2171, avastin, sutent, etc.). This work was deemed to be critical to the success of not only this project, but the PO1 as a whole. Again, Phyllis was urged to take advantage of the unique in vivo systems (Mayo xenograft, in vivo stem cell model) available to us. Cande Manzano, unfortunately, could not attend the RTOG meeting. Cande has described some exciting work on the role of Tie-2 in angiogenesis in malignant gliomas last week. Given the critical role Tie-2 appears to play in angiogenesis, Cande's Tie-2 work should also represent one of the centerpieces of the angiogenesis project. Furthermore, Ken and Monika have identified aquaporin-1 as being a negative predictor in their respective metagene analyses. Of significance, this was the only gene that cross-correlated in both their datasets. Given that aquaporin-1 is known to play a role in angiogenesis, it would be advisable to functionally characterize this molecule and its possible role in mediating resistance to RT+TMZ, as preliminary data for the PO1 grant. Action Items: 1) Phyllis should contact Jann and Howard to investigate the role of the aforementioned signal transduction pathways in enhancing the angiogenic phenotype in tumors using their unique in vivo model systems. 2) The panel of anti-angiogenic agents available to us should be examined in the Mayo panel and Howard's in vivo stem cell model to determine which are most effective and to investigate molecular subtypes of GBM that are most sensitive to anti-angiogenic agents. 3) Phyllis and Cande should communicate about the coordination of their projects. 4) Arnab will share RTOG 0211 correlative data with Phyllis and Cande. 5) Cande should investigate the possible correlation of mesenchymal patterns of gene expression (from Ken's dataset) with the angiogenic phenotype and gene expression patterns associated with response to the various anti-angiogenic agents in stem cells. 6) Given that aquaporin-1 has shown up in several datasets as being of negative predictive value in gliomas and has been reported to play a role in angiogenesis, it would be worthwhile to functionally characterize this molecule in the context of radiation and TMZ resistance, respectively. Stem Cells: Howard Colman described his hypothesis and specific aims in detail. His hypothesis is that over expression of mesenchymal genes (e.g. YKL-40) is associated with increased treatment resistance. Further, he hypothesizes that targeting molecular mechanisms regulating stem cell renewal will improve the therapeutic gain of TMZ+RT in malignant gliomas. Howard has successfully cultured ~ 12 GBM cell lines with stem-cell-like properties and has begun to characterize these cells on the molecular and genetic levels and their relative sensitivities to TMZ. The correlation of TMZ sensitivity to CD133 expression is indeterminate at present. The Chakravarti lab has been able to isolate stem cell-like cells from established GBM cell lines and has found the stem cell-like cells are considerably more radiation resistant than the parent tumor. Likewise, in Howard's panel of stem cell lines, the Chakravarti lab (Palanichamy) has found many demonstrate little or no cell death, even after considerable doses of radiation. There has been no correlation found thus far with regards to CD133 expression and the degree of radioresistance. In fact, some of the more radioresistant stem cells have extremely low expression of CD133! The Chakravarti lab has found variable expression of a panel of 12 stem cell markers in both the MDACC and MGH panel of stem cell lines. There is an ongoing effort to correlate expression of these stem cell markers with the degree of radioresistance. Action Items: 1) Howard will continue to evaluate the relative sensitivities of the MDACC stem cell panel to TMZ, both in vitro and in vivo. 2) Howard will correlate the relative chemosensitivities of these stem cell lines, both to known stem cell-related markers and to the MDACC gene expression data (esp. mesenchymal signatures). 3) There will be active collaboration with the signal transduction, angiogenesis, and DNA repair projects to identify pathways critical to stem cell self-renewal and, hence, therapeutic resistance. 4) The Chakravarti lab will continue to evaluate the relative radiation sensitivities of the MGH and the MDACC panel of stem cell lines, both in vitro and in vivo, using assays identical to Howard's. 5) The Chakravarti lab will correlate the relative radiosensitivities of these stem cell lines, to A) known stem cell-related markers; B) the Harvard and MDACC gene expression datasets and C) to data obtained in the signal transduction and DNA repair projects. Signal Transduction Resistance Mechanisms: The hypothesis of this project is activation of the PI3K/AKT pathway plays a critical role in mediating therapeutic resistance in malignant gliomas through activation of critical angiogenic, DNA repair, anti-apoptotic, and metabolic pathways. The underlying mechanisms by which PI3K signaling regulates angiogenesis will be evaluated by Phyllis and Cande. Likewise, the mechanisms by which PI3K signaling enhances dsDNA repair will likewise be evaluated in the DNA repair project. As part of this project, key PI3K-related molecules and pathways found to be critical in regulation of angiogenesis and DNA repair in preclinical models will be evaluated for their respective prognostic and predictive significance using clinical specimens (RTOG 0525, 9813, and the nondeleted AO trial). The preclinical focus of this project will be to identify mechanisms by which PI3K signaling prevents tumor cell death through suppressing pathways leading to apoptotic and non-apoptotic cell death. Further, the Chakravarti lab has evidence that the PI3K pathway plays a key role in activating metabolic pathways in tumor cells, which are essential for survival. Using the unique preclinical models available to us, these associations will be solidified and exploited for molecular targeting purposes and to investigate novel prognostic/predictive markers using RTOG specimens. Further, there is initial correlative data from RTOG 0211 that activation of PI3K signaling may lead to resistance to targeted therapies such as gefitinib. We shall investigate the associations of constitutive PI3K pathway activation to resistance to other targeted therapies currently under investigation or planned in future RTOG clinical studies. Action Items: 1) Complete correlative analysis of RTOG 0211. 2) Define PI3K-regulated metabolic pathways (Collaboration with metabolomics or Gaby) in preclinical models and cross-correlate with survival endpoints using our clinical correlative data. 3) Define PI3K-regulated survival pathways and devise targeting strategies to enhance the therapeutic gain of RT+TMZ. 4) Investigate the role of PI3K signaling in enhancing survival and self-renewal capacities of GBM stem cells upon exposure to RT. BREAST/TRP Overall Liaison Goals: Facilitate greater incorporation and completion of translation breast cancer projects to increase our understanding of the biology of breast cancer and translate this understanding into better treatment. Promote the visibility of the TRP to successfully demonstrate the importance and effectiveness of this committee on the grant renewal application. 1. Projects on the Table Relevant to TRP Current Concept: Doug Arthur (presenting on behalf of collaborators Henry Kuerer, Bruce Hafty, and translational collaborator Wendy Woodward)-Collection of CTCs as a component of the proposed PBI for IBTR after BCT study. Status: Concept is under review. New Proposal: S. Chmura, (presenting on behalf of collaborator A. Minn) - Feasibility study of SGRT with a translational component including gene array analysis and collection of CTCs. Status: It was discussed the NCI is less enthusiastic about feasibility studies and the main goal of this study may be better propelled as a translational study. Per the TRP chair it should be circulated through the TRP. There is potential this patient population will have sufficient numbers of CTCs to permit array analysis of these cells rather than just from blocks. In a large number of patients with attention given to breast cancer subtypes this would represent an enormous improvement on the depth of data recently published in the NEJM by M. Clarke's group. Plan: I will touch base with Andy Minn, Steve Chmura, Adam Dicker and the interested members of TRP to generate a plan for this that can be applied to any applicable protocol as well as to review and potentially optimize the translational goals for the proposals on the table. Much of this literature has been spearheaded in the field of breast cancer and my hope is the breast group will be at the forefront of exploring the clinical relevance of the preclinical findings. In particular I recommend choosing a method of analyzing circulating tumor cells will allow for the characterization of additional markers on these cells including stem cell markers. This is an area the TRP is very interested in and there is collective experience through the TRP membership. We have access to expertise and technology through my participation in the MD Anderson Advanced Research Center for Micrometastaic Disease (ARC-MD) chaired by Massimo Cristofanilli (Cristofanilli et al, NEJM) in collecting and analyzing this material. In addition, per Dr. Chmura, University of Chicago is looking into purchasing this technology, and Dan Hayes (senior author on the above paper) has been in touch with Adam Dicker via SWOG and is interested in getting involved in RTOG). Others may have it as well and this information will be gathered. 2. Breast TRP Meeting during Summer Meeting 2007: We will plan to have a separate breast TRP meeting at the next RTOG meeting to allow for more time to discuss technical considerations and translational goals. Ideally this will occur prior to the main Breast and TRP meetings so these ideas can be subsequently be shared with the entire group. In particular, I want to be sure all are aware of what tissue has been collected on behalf of RTOG and where it is stored. Currently none of it is in the RTOG tissue repository. The TRP is exploring what the path is to ask questions using this material and we hope there will be proposals put forth to use this material. We can also discuss standardizing non-invasive materials collected for future TRP use that are not currently being collected, for example urine or oral mucosa scrapings. Action Item: If time permits I would like a few minutes on the breast committee agenda to update the group on the results of the TRP breast meeting. 3. Stem Cell Symposium: The TRP has discussed holding a symposium at the 2/08 RTOG meeting and Cancer Stem Cells was selected from a number of suggestions as the focus. I will be working to coordinate all who have an interest or expertise in this area to put forward an agenda and contact speaker for this. The objectives will be to evaluate how this area of research can be incorporated into clinical trials across sites, and to educate the membership as to the state of the art. We will plan to have the talks from the symposium transcribed so that a summary statement can be generated from the discussion. In preliminary discussion it has been suggested that this be published as a report from the TRP rather than from specific authors, but this is open for consideration. Dr. Cox, the editor of the Red Journal has preliminarily committed space for this endeavor. 4. Breast Atlas: Although currently the primary focus of this is clinical, for the purpose of the grant renewal application this will represent a rich opportunity to highlight the translational physics projects may be possible based on current strategies and plans. Action Item: We may want to consider identifying someone with an interest in this beyond the clinical implications to both coordinate translational physics ideas as they relate to the RTOG breast group and importantly to summarize and write this component of the renewal. GENITOURINARY/TRP John Petros, MD: Gave a 10 minute talk reviewing new nanoparticle technology for measuring molecular markers, 5-10 at a time on tissue sections. Collaboration using samples from RTOG 8610. Should organize teleconference to discuss which markers to test and logistics HistoRx: Access to this technology uncertain. No formal plan in place. Need to review strategy in teleconference. Prioritizing of Markers for RTOG 9408 Arul C. Project was met favorably. Restrict Mark Ritter analysis to 3 markers for now: Ki67, p53 and MDM2. Hold on bcl-2/bax, Cox2. This is very unfortunate, since this is a funded. R01 and will certainly have an impact on future RTOG-based grants. Debate about a Cox2 Inhibitor-Based Trial Adam Dicker: No encouraging data in RT trials. However, Cox-2 inhibition has activity in prostate cancer. Concern about cardiac deaths and litigious environment. Whether this should prevent a trial was discussed. Biomarker Panel from RTOG 9202 High priority was given for RTOG statisticians to work on developing a panel, as has been done in breast cancer is funded through a Tobacco grant. Javier Torres-Roca would like to participate in this process. Testing of the results in RTOG 9413 was approved. RTOG 0612 Everyone was encouraged to participate. Since RTOG 0232 may close, there was discussion about amending the protocol or developing an independent protocol for EBRT patients. Prospective Biopsies from RTOG EBRT Protocols Jeff Michalski volunteered to work on this. Imaging component: MRS, DCE (optional?) - Pre & Post Treatment Prostate biopsy at time of fiducial marker placement - Discuss with Arnab to see if he is OK with this Overlaps in many ways with RTOG 0612 RTOG 0612: Prostate Microarray Study Model for IHC studies: Chromavision vs. HistoRx Update on protocol RTOG 8610 markers Update on RTOG 9202 markers Update on RTOG 9408 markers Ritter Grant progress Arul Chinnaiyan, Spore Project ProposalTMPRSS: ETS fusion protein variants correlates with response to RT, RT+ADT or initial PSA decline Grants/Funding/New Projects John Petros/Peter Johnstone: Multiplexed Nanoparticle technology for biomarker assessment Multiple markers in RTOG 9408: Ritter R01 E2F-1/Mad2 in RTOG 9202: Pollack R01 pending Survivin in RTOG 9202: Chakravarti R01 pending mRNA gene expression RTOG 9413: Pollack R21/part of R01 Possible project for P01 mRNA gene expression frozen tissue RTOG 0232: Chakravarti Possible R01 vs. project for P01 Update on Bladder Protocol Markers Bill Shipley: VEGF study New Directions/Business New protocols Imaging concepts: Post-treatment MRI spectroscopy Based on UCSF data Cox2 trial Markers for RTOG Protocol 8610 Analyses Completed (Papers Pending) P53: Published JNCI (Grignon) DNA ploidy: Published JCO (Pollack) Ki-67: Published Clin Cancer Res (Pollack) p16/pRb: Published, JCO (Chakravarti) MDM2: Published, Cancer (Khor) Androgen receptors: Pubished AJCO (Wahab) Bcl2/Bax: Published Cancer (Khor) Cox-2: Paper submitted status? (Hughes, Dicker) Stat3: Urology, in press (Torres-Roca) Survivin: ASTRO 2005, Paper in Prep (Chakravarti) AKT: ASTRO 2005 (Chakravarti) PKA: Paper In Prep (Khor) Markers Proposed for RTOG Protocol 8610 with Analyses being completed (Papers pending) - NFKB: ASCO 2006 VEGF-D: Proposed (Coen) pMAPK Proposed (Chakravarti) P27: Proposed (Teh) GSK Proposed (Teh) Forkhead Proposed (Teh) IGF: Proposed (Konski) EGFR: (Chakravarti) Prostate RTOG Protocol 9202 Publications/Presentations (New) Ki-67: Published in JCO (Pollack) MDM2: ARS 2005 Abstract (Pollack) Multi-marker (MDM2/p53/Ki-67): ASTRO 2005 (Pollack) p53 and Ki-67 ACIS data complete Paper planned P53: Paper In press IJROBP (Che/Grignon) P16: Paper resubmitted (Chakravarti) Cox2: ASTRO 2006 Paper at RTOG (Khor & Dicker) Bcl-2/Bax: ASTRO 2006 Paper Submitted (Khor) Prostate RTOG Protocol 9202 Planned Analyses Hierarchical clustering?: Multimarker study E2F-1/MAD2: Planned (R01 Submitted, Pollack) Survivin: Planned (R01 Submitted, Chakravarti) Androgen Receptor? Ritter R01 Based On RTOG 9408 RTOG 9408 (Ritter & Pollack) R01 Funded Ki-67 complete p53 in progress Planned: MDM2, bcl-2/bax Analysis anticipated soon? - survival endpoint Pollack Tobacco/R21: RTOG Protocol 9413-Pollack (Hagg): Proposed R21 Patient group similar to RTOG 9202 Confirm IHC markers from RTOG 9202 Blocks have not been cut yet Ideal for mRNA gene expression profiling Focus on: Haqq: Genes associated with nodal mets Pollack: Apoptotic pathway Builds on our immunohistochemical experience in RTOG 9202 SARCOMA/TRP Dr. John Kane presented problems with completing the micro array study RTOG 9514 and MGH study. Issues relate to availability of tissue for study. Additional material will be sought. No funds from grant have been expended. Dr. Dina Lev from MD Anderson Cancer Center presented and extended report of work being done at there. She also presented at Sarcoma Committee and RTOG TRP meeting. An extensive number of multidisciplinary studies are being done at MD Anderson Cancer Center addressing the biology of sarcomas and potential methods of management. Dr. Burton Eisenberg from Dartmouth Hitchcock Medical Center reported the status of the neoadjuvant STI-571 trial. The trial has been completed and data is being collected and analyzed. He hopes to have a manuscript ready for consideration by June 2007. Dr. Adam Dicker, Chairman of the RTOG Translational Research Program attended a part of the meeting. He asked the Sarcoma TRP Committee to initiate a tissue, blood, buffy coat, etc. collection only protocol. Considerable discussion concerning this occurred.