Presentation on theme: "Tumor Microenvironment Network (TMEN) Division of Cancer Biology"— Presentation transcript:
1Tumor Microenvironment Network (TMEN) Division of Cancer Biology Dinah Singer, Ph.D.DirectorSuresh Mohla, Ph.D.TMEN Program DirectorDivision of Cancer Biology1. Although this is a reissuance, its goal is not simply to continue the existing Network, but to build on and extend the success of the original TMEN.2. present our vision for the new TMEN, which will highlight its differences from the existing one.
2TMEN : GoalsGenerate a comprehensive understanding of the composition of the normal stroma and the role of the stroma in tumor initiation, progression and metastasisDevelop resources and infrastructure critical to the broader research community to advance understanding of the tumor microenvironment (TME)When TMEN was first established in 2006, it was already understood that there is a complex interplay between the tumor and its surrounding stroma.However, the composition of the stroma and its role in cancer remained unclear. Although a number of investigators – including some at this table – were making significant progress, it was clear that the advances in the field could be dramatically expedited through a programmatic effort that:importantly, working together to:Generate the approaches and critical reagents and resources, such as stromal cell markers, that would
3TMEN 2006-2011: Current Program Nine interdisciplinary groups characterizing the TME in major cancer sites, emphasizing human samplesIdentifying and translating promising leadsCollaboratively addressing the complexity of TME by assessing:The co-evolution of tumor-associated stromal cell types and the tumorStromal cell interactions with each other and with tumor cellsGenerating novel reagents, models and technologies for the research communityThe success of the TMEN, as a network has derived from the collaborative efforts of the Network to address the complexity of the TME and to generate novel resources for the community.Neither of these could have been accomplished through traditional funding mechanisms and depended on the Network infrastructure.
4TMEN 2006-2011: Areas of Emphasis MicroenvironmentInitiation Progression MetastasisTumor initiating cellsTumor heterogeneityStromal cell types and cellular processesComplex signaling pathwaysGenes and geneticsThe first TMEN RFA identified 5 broad scientific areas of emphasis for which a collaborative, network approach would give us insights into the role of the microenvironment in initiation, progression and metastasis.In the 4 years since the TMEN was established, significant progress has been made in each of these areas, much of it deriving from TMEN-supported research.In the next few slides, I’ll highlight just a few of the accomplishments of the TMEN. However, a more detailed list can be found in the concept and other documentation that you’ve been provided.4
5TMEN 2006-2011: Scientific Accomplishments Tumor Initiating Cells:Combined invasive gene signatures in tumor initiating cells and stromal wound repair response genes robustly predict metastasis-free and overall survival in breast, medulloblastoma, lung and prostate cancer patientsNew therapeutic approaches to circumvent the chemo- and radio-resistance of tumor initiating cellsStromal Cells:Delineation of mechanisms to describe the roles of the various bone marrow-derived cell lineages in tumor growth, invasion, inflammation, angiogenesis and metastasisStromal genes and factors:Cancer-associated stromal gene signatures predict progression to biochemical recurrence in human prostate cancerCancer-associated neurogenesis in human prostate predicts aggressive diseaseUnderstanding the signaling and metabolic pathways that distinguish TIC’s has allowed new therapeutic approaches.Targeting the Notch ligand Delta can reverse chemoresistant of CSCs. Phase I trial initiated.Low ROS levels account for redioresisitance of CSC and inhibitor studies showed that reversal may make CVSC radiation sensitive.5
6TMEN 2006-2011: Scientific Accomplishments Tumor Initiating Cells:Combined signatures of invasive and wound repair response genes in tumor initiating cells robustly predicts relapse and overall survival in breast, medulloblastoma, lung and prostate cancer patientsNew therapeutic approaches to circumvent the chemo- and radioresistance of tumor initiating cellsStromal Cells:Delineation of the mechanisms by which bone marrow-derived cells affect tumor growth, invasion, inflammation, angiogenesis and metastasisCancer-associated reactive stromal volume predicts progression to biochemical recurrence in human prostate cancerCancer-associated neurogenesis in human prostate predicts aggressive diseaseTumor microenvironment signature of metastasis (TMEM), which includes tumor cells, TAMs and endothelial cells, accurately predicts systemic metastasis from primary breast cancer samplesExamples of some of the seminal studies that have been published in high profile journals – commonality of stem cell markers across many tumor sites.Targeting the Notch ligand Delta can reverse chemoresistant of CSCs. Phase I trial initiated.Low ROS levels account for redioresisitance of CSC and inhibitor studies showed that reversal may make CVSC radiation sensitive.6
7TMEN 2006-2011: Programmatic Accomplishments Scientific CollaborationsIdentification of human tumor initiating cell fate using intravital imagingCharacterization of human prostate stromal changes, both genetic and epigeneticDevelopment of imaging beacons to determine role of specific metalloproteases in tumor and stroma
8TMEN 2006-2011: Programmatic Accomplishments Scientific CollaborationsIdentification of human tumor initiating cell fate using intravital imagingCharacterization of human prostate stromal changes, both genetic and epigeneticDevelopment of imaging beacons to determine role of specific metalloproteases in tumor and stromaCritical Resources DevelopedValidated EHS sarcoma matrix (“Matrigel”) resourceStromal antibodies – validation of existing antibodies and development of novel antibodies for stromal and tumor initiating cellsTransgenic mice expressing stromal TVB receptors for targeting with RCAS vectorsBank of tumor initiating cells derived from early passage human tumors of head and neck, breast, colorectal cancersBank of murine bone marrow cells, for generating BMDCA major effort in the Network has been the development of resources which are available to the general research community. A comprehensive list is provided at the end of the slide set in your books.These resources have contributed to the rapid progress that’s been made in TME research and are available to the research community.8
9Proposed TMEN Reissuance Concept: Why? Investigation of emerging novel concepts in TME will require new teams of interdisciplinary scientists to collaborate on large scale, complex problemsNetwork structure will:Accelerate the development of new areas in TME, and of resources for the research community. (Investigator-initiated mechanisms are not suited for such efforts.)Provide a focus for interacting with other NCI programs, to promote interdisciplinary effortsEncourage established investigators to study the TME, through the collaborative U01 programFacilitate training of junior investigatorsIn short, in its first 4 years, TMEN has either achieved or made significant progress, in all of the goals laid out in the original RFA.So, why are we requesting a reissuance rather than allowing the field to develop further through investigator-initiated research mechanisms, such as RO1’s or PO1’s?During the past 4 years, and in large part as a result of the advances made by TMEN in our understanding of the TME, a number of novel concepts have emerged that will require new approaches and expertise and new teams of researchers to collaborate to unravel the complexity of the TME.A network structure will not only enhance the ability of interdisciplinary groups to make rapid progress, but will also provide leadership within the field and ensure the generation of the new resources and reagents that will be critical for the entire research community to develop these novel areas.Neither of these can be achieved through classical investigator-initiated mechanisms.expertise of a wide range of many disparate fields. Importantly, a network forms an organizational structure and focal point for interacting with other NCI funded programs, such as ICBP, MMMHC, phys-onc, nanotech, edrn, imaging.
10TMEN Reissuance: Areas of Emphasis MicroenvironmentInitiation Progression Resistance Metastasis Recurrence- Tumor initiating cells- Tumor heterogeneity- Stromal cell types and cellular processes- Complex signaling pathways- Genes and geneticsStem cell nicheMetastatic nicheTumor dormancy and metastasisMetabolic dysregulationCell fusion and exosome secretionMicrobiome (bacteria; viruses)Gradients and flow of soluble factorsreissuance will explore a range of novel concepts that emerged from studies over the past 4 years and span the cancer continuum; these now include resistance and recurrence, in addition to initiation, progression and metastasis.The Network will provide the infrastructure to ensure that the needs of the community in these areas are met.Emerging Areas10
11Proposed TMENOpen RFA competition emphasizing emerging areas, soliciting new interdisciplinary teamsUp to 9 Research Programs (U54 Consortia)Continued support at current total level of $9.08M/yearFive year durationBecause of the changes in emphasis, we anticipate that there will be considerable turn-over in the TMEN groups and their composition.In conclusion, our intent in requesting this reissuance is not to simply continue the current TMEN. It is to build on and significantly extend the progress that was made by the existing TMEN.
14TMEN Grant and Network Activities Funding FY2006-2010 TMEN Project and Network ActivityAverage Total Cost per YearIndividual Grant Support9 U54’s(5 fully funded, 4 partially funded)$8,401,355Network ActivitiesResource developmentCollaborative projects$ ,033Total Cost per year$9,085,388
15Current TMEN Principal Investigators Mina Bissell (LBL, UC Berkeley)Michael Clarke (Stanford)John Condeelis (Albert Einstein)Eric Holland (Memorial Sloan Kettering CC)Richard Hynes (MIT)Lynn Matrisian (Vanderbilt)Stephen Plymate (Univ. Washington)David Rowley (Baylor)Timothy Wang (Columbia)
16U of Washington/ Fred Hutch TMEN: Collaborative NetworkMITVanderbiltAlbert EinsteinColumbiaStanfordBaylorMSKCCU of Washington/ Fred HutchLawrence BerkeleyDana Farber CoreCollaborative U01 Dana FarberCollaborative U01 VanderbiltReagentsModelsProtocolsCollaborative U01 U Penn
17TMEN 2010-2015: Examples of Scientific Goals Using multi-disciplinary approaches , delineate biological mechanism(s)governing the TME interactions which are critical for tumor development,progression and metastasis, and therapeutic resistance and tumor recurrenceFocus on specific tumor sites, using human cancer tissues and modelsCharacterize component cells and matrix molecules in normal organ and tumor-associated stroma; examine alterations in the microenvironmentIdentification of lineages of bone marrow-derived cells and immune cells and their effects in tumor initiation, progression and metastasisIdentify and characterize the stem cell niche; origin of stromal cellsCharacterize the role of microbiome (bacteria and viruses)Characterize the role of metabolic dysregulation in TMEDelineate the functional relevance of tumor cell-host cell fusion and exosomesDetermine the effects of cancer therapy on the TMEIdentify and characterize the role of TME in therapeutic resistanceIdentification and characterization of gradients and flow of soluble factors
18TMEN Evaluation Criteria The following broad criteria will be used to judge the success and effectiveness of the proposed programSuccessful identification of novel components of the normal and tumor stroma in primary and metastatic tumorsNew insights into the composition, role and origin of tumor-associated stroma during cancer initiation, progression and metastasisDetermining the effects of cancer therapy on the tumor microenvironmentConsortium participation in collaborative activities aimed at improving existing technologies and developing novel reagentsDevelopment and effective dissemination of resources to the research community
19Independent Program Evaluation Report Addressed goals of the original RFAEvaluation based on literature, NCI grants documentation, interviews and staff interactionSeparate evaluations from outside expertsReviewers’ comments reflect their overall enthusiastic endorsement“Overwhelmed by the accomplishments of the groups associated with TMEN- both in terms of number of critical questions being addressed, the quality of the investigators and the innovative approaches being pursued.”“if research in this area was funded entirely through the R01 mechanism, there would only be a trickle of isolated research projects in the area ““The coordination of resource generation has been largely driven by staff.”“the program has exposed investigators who were working in select areas of the microenvironment to think ‘bigger’Evaluation based on information generated from multiple sources
20Current Portfolio Analysis TMEN is the only funded program addressing a comprehensive approach to understand the TME, and its critical role in mediating tumor progression, metastasis and therapeutic resistanceThe total TME portfolio budget in NCI is $43M. DCB’s portfolio includes the current 9 TMEN U54s, 3 U01s, and 61 investigator-initiated grantsVery few grants focus on the new areas proposed in this reissuance. Most examine cell migration, proteases, angiogenesis, hypoxia or metastasisInvestigator-initiated grants are unable to pursue comprehensive studies to address the microenvironment using “tumor-as- an-organ” approach as these mechanisms are not suited for to these types of studies
22BMDC bankBone marrow cells will be isolated from two commonly used strains of mice. These are:(a) the Rosa26 (C57BL/6Jtrosa26) mice (b) smooth muscle a-actin promoter driving a reporter (EGFP)RCAS Vector development targeting stromal cellsa) two RCAS(A) and two RCAS(B) vectors were made. Just to clarify, these are the avian virus vectors that infect the respective transgenic mice that express the RCAS (A or B) receptors called TVA or TVB. Two TVB and one TVA mouse model have been generated(i) FSP1-TVB mice (S100A4, that is expressed in many cells of mesenchymal origin including fibroblasts).(ii)SM22-TVB mice (expressed in smooth muscle cells).Currently the founders are being characterized at Vanderbilt – they have already been initially screened by Lionel Feigelbaum (sp?)b) The third mouse (started late in the contract) is the "hit and run" mouse that allows for expression of TVA upon crossing with any Cre mouse. That means one can target expressing either stromal or epithelial cell type.c) The RCAS are avian retroviruses that can only infect cells that express the appropriate viral receptor
23Standardization of Antibodies and generation of novel antibodies Antibodies used for flow cytometry isolation of cells - sent to NCI repositoryfor distribution to other TMEN laboratories and the broader NIH research communityFLOTILLIN-2/ESA FITC MAB 29HU CD10 PE-CY5 MAB HI10AHU CD140B BIOTIN MAB 28D4HU CD166(ALCAM) PEHU CD18 PE-CY5HU CD2 PE-CY5HU CD20 PE-CY5 MAB 2H7HU CD24 FITC MAB ML5HU CD24 PE MAB ML5HU CD3 PE-CY5 MAB UCHT1HU CD31 BIOTIN MAB M89D3HU CD31 BIOTIN MAB WM59HU CD44 APC MAB G44-26HU CD45 PE-CY5 MAB HI30HU CD45 PE-CY7 MAB HI30HU CD64 PE-CY5 MAB 10.1HU CD66 FITCHU CD66 PEHU EGFR FITC (0.05MG)HU EGFR PEMS CD140A BIOTIN MAB APA5MS CD90.1(THY1.1) APCMS H-2KD BIOTIN MAB SF1-1.1SAV PE-CY5SAV PE-CY7
24List of cell lines generated for antibody target overexpression PC12 parental cell linePC12 vector control cell linePC12 Laminin A4 overexpression cell line293T lysates from overexpression of TGFβ1,β2 and β3 as well as Cercam and Leprel 2 have been generated and used for Elisa and western blot screening of antibodies.List of stable knockdown cell lines for antibody targetsMCF7 GFP shRNA KD cell lineMCF7 TGFβ1 KD cell linesMCF7 TGFβ2 KD cell linesMCF7 TGFβ3 KD cell linesMDA435 GFP KD cell lineMDA435 Leprel 2 KD cell linesMCF7 CerCam KD cell lines and GFP KD control lineT98G GFP KD cell lineT98G Laminin Alpha 4 KD cell lines
25AntibodiesmAb against CercammAb against Laminin Alpha 4antibody supes against TGF β1/2/3 as well as Leprel 2REAGENTS AND PROTOCOLSProtocol for separation of mouse stroma from human tumor cellsProtocol for separation of human epithelial tumor cells from non-tumor componentsProtocol for LCM following by high-resolution oligo array-CGHProtocol for optimized WGACustom Agilent CGH 2x415K oligo microarray, optimized for resolution vs. costMonoclonal Antibodies against Laminin α4 and CercamTumor initiating cells derived from early-passage human xenograft cancer tumorsestablished from cells isolated directly from patient tumors - sent to the NCI.Colon cancer - 9 xenograft tumorsBreast cancer - 5 xenograft tumorHead and neck cancer - 10 xenograft tumors
26TMEN Publications: 2006-2010 Year Number of Publications 2010 18 In prep (2010) 9Total