Olivera J. Finn - US grants

Analyses of human effector T cell repertoire in renal allograft rejection indicate that responses to a particular alloantigen, or a number of alloantigens, do not utilize all the available repertoire, but that some T cells have a selective advantage giving rise to predominant clonotypes. The advantage appears to be a result of an expression of a particular T cell antigen receptor beta chain gene. Using T cell lines established from lymphocytes infiltrating a rejected allograft, we propose to clone and sequence the predominant TcR beta chain gene to understand the molecular basis for their selection. The long term goals of this project are to use this knowledge to design ways of specifically eliminating or down-regulating the selected clonotypes in order to achieve prolonged graft survival. The immediate benefit of this project will be a greater understanding of the relationship between the structure of the T cell antigen receptor and its function in the recognition of antigen/MHC or foreign antigen alone.

It has now been successfully shown in a number of animal and human tumors that a specific immune response can be generated against the tumor. This foreshadows an era of immune manipulation of tumor growth and eventual recruitment of the immune system in the process of tumor destruction. The success of immunotherapy depends on further understanding of the multitude of reasons why the observed anti-tumor response is seldom effective. At the root of that understanding is the need to know the target antigen on the tumor cell. Patients with pancreatic and breast adenocarcinomas mount a specific T cell response to their tumors. CD8+ as well as CD4+ T cells can be isolated from tumor draining lymph nodes, both populations specifically recognizing epithelial mucin as the target antigen. This antigen coats the tumor cells and is also present on normal ductal epithelial cells but not recognized by the T cells until malignant transformation unmasks specific T cell epitopes. The epitopes recognized by the two T cell populations reside on a 20aa tandemly repeated peptide which comprises the mucin polypeptide core. The tandem repeat structure endows this antigen with antigenic multivalency, potentially useful in immunotherapy, which gives it the ability under certain circumstances to activate T cells directly. The ability of this antigen to stimulate both helper and cytotoxic T cells fulfills a critical requirement for establishment of an efficient immune response. Yet, the patient succumbs to the tumor. Potential reasons may be insufficient numbers of tumor-specific CTL and anergy in the helper T cell compartment. In this application we propose: I. To investigate MHC-restricted and MHC-unrestricted activation of CTL by mucin antigen in order to validate methods for in vitro and in vivo generation of large numbers of CD8+, tumor specific CTL. II. To investigate routes of antigen presentation which can lead to generation of large numbers of functional CD4+, tumor specific T helper cells. III. To investigate the state of anergy of the CD4+ T cells and ways in which this anergy may be broken, or the cells otherwise altered to provide functional help. The long term goals of the project are to fully understand the anti-tumor mucin response and its potential for immunotherapy.

Breast adenocarcinomas arise from malignant transformation of normal ductal epithelial cells. In the process of characterizing specificity of T cell lines derived from breast cancer patients and cytotoxic for breast tumors, we identified ductal epithelial cell mucin as the antigen on the tumor capable of stimulating patients' T cell immunity. We further identified one epitope on this antigen which serves as a target for tumor specific CTL. Normal mucin producing cells do not express this epitope and its presence correlates with the susceptibility to lysis of tumor cells by the CTL. This epitope and several others are preferentially expressed on tumor mucin due to incomplete glycosylation of this molecule by the tumor. We have determined that transfection of EBV immortalized B cells or primary fibroblast cultures with mucin cDNA plasmid expression vector and treatment of transfected cells with inhibitors of O-linked glycosylation results in tumorlike processing of the mucin molecule and high level expression of these epitopes. The overall goals of this proposal are to utilize the knowledge of one tumor specific epitope we have identified so far on breast cancer mucin, capable of stimulating human T cells, and our ability to recreate this epitope, as well as several others, in mucin transfected normal cells, to explore the possibility of constructing breast tumor specific vaccine. Sequence identity between the chimpanzee and human mucin allows us to test this mucin-based vaccine in vivo. 1) We will utilize plasmid expression vectors to express mucin in autochthonous antigen presenting cells(APC), (human as well as chimpanzee), and glycosylation inhibitors to expose tumor specific mucin epitopes, and perform in vitro analysis of T cell responses to these epitopes. 2) We will test the ability of glycosylation-inhibited, mucin transfected autochthonous APC to stimulate specific T cell responses in vivo, in chimpanzees immunized with these cells. 3) We will construct and test in vitro and in vivo new expression vectors containing partial mucin polypeptide core sequences with glycosylation sites mutated in order to ensure incomplete glycosylation and constitutive expression of tumor specific mucin epitopes.

Maintenance of antigen specific T cells in vitro requires periodic stimulation with specific antigen presented by appropriate APC, and the presence of IL-2, T cell growth factor. An early result of T cell antigen receptor (TCR) occupation by antigen is the rapid hydrolysis of membrane phosphatidylinositol 4,5-bisphosphate, liberating the second messengers inositol trisphosphate and sn- 1,2-diacylglycerol. Diacylglycerol, in synergy with the increase in intracellular calcium induced by inositol triphosphate, activates the serine-threonine-specific protein kinase C (PKC). We have previously shown that constitutive expression and activation of PKCgamma in fibroblasts results in increased proliferation rate in vitro, increased saturation density and lowered requirement for growth factors. This prompted us to constitutively express PKC in antigen specific T cell clones in order to test its growth promoting effect on these cells, and to explore the possibility that T cells might behave as if their antigen receptors were perpetually stimulated by antigen. T cells which constitutively expressed PKC had stable high level expression of IL-2 receptors, increased proliferation rate, increased longevity in vitro, and complete independence of antigen, while maintaining strict antigen specificity and effector function. In the studies proposed here we plan to: I. Use newly constructed amphotropic retroviral vectors to transduce PKCgamma, as well as alpha and beta genes, into human anti-tumor effector T cells and evaluate in vitro what effect constitutive overexpression of these genes has on proliferation, longevity and anti-tumor effector function of these cells. II. Use PKC transduction to facilitate isolation of tumor specific T cell clones from tumor patients peripheral blood, lymph nodes or tumor samples. III. Use a well established animal tumor model of adoptive immunotherapy to test the effect of PKC transduction into T cell lines and clones used to mediate immunotherapy. IV. Construct new retroviral vectors which contain a suicide gene capable of destroying the transfected effector cells, once they have accomplished their immunotherapeutic function, for future application of this method in human clinical trials of adoptive immunotherapy of cancer.

DESCRIPTION (provided by applicant): Epithelial cell glycoprotein MUC1 is overexpressed and hypoglycosylated on all human adenocarcinomas and is recognized by the immune system as a tumor antigen. In basic and preclinical studies supported by the long-standing grant that this application aims to renew, we have deciphered numerous interactions of MUC1 and the adaptive immune system and this knowledge has been translated to MUC1 based vaccines for pancreatic, breast, colon and prostate cancer. We also made an unexpected observation that MUC1 interacts directly with the cells of the innate immune system and thus might play an important role in inflammation and cancer. Our hypothesis is that overexpression and aberrant glycosylation of MUC1 promotes first chronic inflammation and later progression to cancer and thus MUC1 should be targeted not only for tumor therapy, as studied so far, but also for therapy of chronic inflammation before tumors develop. We propose two specific aims: Specific Aim 1. To define MUC1 related cellular and molecular events that regulate (induce, promote) chronic inflammation and cancer. We will use the new mouse model developed in the previous grant period as well as the information we obtained regarding the novel interactions of the MUC1 glycoprotein with the innate and the adaptive immune system. Specific Aim 2. To test the ability of MUC1-specific or MUC1-related immunotherapy to prevent chronic inflammation and cancer. We will use vaccines based on immunogenic MUC1 glycopeptides defined in the last grant period as well as MUC1 specific TCR transgenic mice developed in the same period. We have preliminary evidence that inducing MUC1 specific adaptive immunity can prevent not only cancer but also chronic inflammation. Nobody has yet considered MUC1 as a target for therapy, and specifically immunotherapy, of chronic inflammatory diseases that carry increased risk of cancer. PUBLIC HEALTH RELEVANCE: This is a long-standing project directed towards immunotherapy of cancer and cancer vaccines based on the tumor antigen MUC1. In this renewal application, work will be done to show that this form of immunotherapy and the same vaccines may be useful not only for cancer but also for treatment and prevention of chronic inflammations that increase risk for cancer.

Aberrant glycosylation of the mucin molecule (MUC-1) expressed on epithelial tumors leads to the exposure of novel tumor-associated core protein epitopes which are recognized by tumor specific antibodies and cytotoxic T cells (CTL). Consequently, MUC-1 mucin could be considered a possible target for tumor immunotherapy. A candidate anti-mucin cellular vaccine employing autologous dendritic cells instead of tumor cells for the presentation of tumor associated mucin epitopes was tested in chimpanzees because they express the same molecule with the same sequence and tissue distribution. In preparation for the in vivo studies we obtained blood samples from various animals at the time they were undergoing routine physical examinations. We developed a culture system to grow chimpanzee dendritic cells in vitro (ref. 1). This system was then tested in vivo. First we examined the migration potential of in vitro grown DC and found that they migrate appropriately to the n earest dr aining lymph node where they contact T cells and maintain their immunostimulatory phenotype (ref.2). Dendritic cells were then grown for six days in vitro and then loaded with a mucin synthetic peptide or a control antigen ovalbumin. Autologous antigen loaded dendritic cells were then injected IV, and animals boosted once with antigen in conventional adjuvant TiterMax. Three weeks later blood and lymph nodes were collected and examined for the development of mucin-specific or ovalbumin-specific immune response. We found that dendritic cells were effective at inducing OVA-specific immunity, but not much better that the conventional adjuvant (ref. 3). We saw no immunity generated to the mucin peptide with either regimen, confirming our observations in vitro that this peptide sequence does not contain helper epitopes. We have done another study using the same Muc1 immunogen, but another adjuvant, LeIF, known to promote cellular immunity. We did find helper cell responses, IFN-gamma production and Ctl responses elicited in the immunized chimpanzees (ref. 4) FUNDING NIH/ R01 CA57820 $166,648 7/01/92 - 6/30/98 K01 RR00119 $ 56,650 7/01/96 - 6/30/01 PUBLICATIONS Barratt-Boyes, S.M., Kao, H. and Finn, O.J. Chimpanzee dendritic cells derived in vitro from blood monocytes and pulsed with antigen elicit specific immune responses in vivo. J. Immunother. 21:142-148, 1998. P51RR00165-38 1/1/1998 - 12/31/1998 Yerkes Regional Primate Research Center

vaccines; neoplasm /cancer; immunology; lymphatic system; Primates; Mammalia;

The goals of this proposal are to create a tumor antigen discovery system using human dendritic cells as antigen presenting cells and the most recent technological advances in in vitro culture of human T cells, to attempt to identify candidate tumor specific antigens. This is a novel approach in as much as all previous work in this area has been utilized secondary responses of T cells from cancer patients and tumor cells as antigen presenting cells. Recent evidence from several investigators point towards profound defects in cancer patients' T cell function, making this source less than optimal for defining potential tumor targets. We hypothesize that using naive T cells from healthy donors and a new, powerful in vitro priming system dependent on dendritic cells, should bypass the problem of tumor induced immune suppression in vivo and uncover all potential tumor antigens which have remained undetected in the recall responses. We will focus on lung cancer for which no specific antigens have been identified and on HLA-A2 restricting element, the most common human Class I allele. Emphasis will be placed on the ability of peptides, naturally processed by the tumor, to prime CD8+ T cell responses in vitro, when presented by dendritic cells. In addition, whole tumor proteins will be fractionated and then processed and presented by the dendritic cells to explore their potential of priming tumor-specific helped CD4+ T cell responses in vitro. One tumor antigen, MUC-1 mucin, that we have identified on breast and pancreatic adenocarcinomas is laos expressed by lung adenocarcinomas. Thus in addition to identifying new lung tumor antigens, we will test the full potential of lung tumor mucin to stimulate CD8+ and CD4+ T cells when presented by dendritic cells. Specific aims are: 1. To use dendritic cells as sensitive indicators of Class I restricted tumor specific that can prime CD8+ T responses in vitro. 2. To allow dendritic cells to take up and process tumor cell proteins and indicate those that can prime CD4+ T cell responses in vitro. Having shown the utility of this method, the ultimate goal of these studies will be to use the identified peptides and proteins for designing lung cancer vaccines or facilitating adoptive T cell therapy of lung cancer.

neoplasm /cancer immunology; neoplasm /cancer therapy;

Immunological solutions to cancer problem lie in our ability to elicit effective, potent immune responses, with life long memory, against antigens which are not much different then self molecules. The problem is compounded by different and often immunosuppressive effects on the immune cells by tumor cells. This exciting field of research continues to provide challenges and opportunities for young scientists well trained in the newest paradigms in immunology. The immunology faculty at the University of Pittsburgh School of Medicine have successfully trained many talented investigators in the past who have already made significant impact in the field of tumor immunology. We propose to continue to recruit the best and the brightest into this area of research through a predoctoral and postdoctoral training program in Cellular and Molecular Mechanisms of Tumor Rejection. We will train our students and fellows within our Graduate Training Program in Immunology, one of several training programs of the Interdisciplinary Training program in Biomedical Sciences of the University of Pittsburgh School of Medicine. The goals of the Training Program are to provide students and fellows with courses, research projects, highly qualified mentors and financial resources which will allow them to: 1. Elucidate the basic mechanisms of initiation of tumor-specific immunity; 2. Define the effector mechanisms in the anti-tumor immune response; and 3. Understand the requirements for establishment of long term anti-tumor immune memory. This will be achieved through well planned and organized course activities, combined with program activities which promote substantive communication among the program faculty and trainees. The program will also strive to train both graduate students and fellows in other important aspects of their profession: 1. Reporting in a timely fashion research results in peer reviewed manuscripts and at national and international meetings. 2. Writing grant proposal to various private and government funding agencies. 3. Teaching in courses related to their field of expertise.

DESCRIPTION (provided by applicant): The goals of this revised renewal application are based on the new information we acquired in the previous funding period and the overall accumulation of knowledge in the DC field in general. Seven years ago, the state-of-the-art knowledge divided DC into two subsets, lymphoid and myeloid. Within each subset, they could exert either immunogenic or tolerogenic effects, depending on the state of maturation and the ability to provide to naive T cells antigenic and co-stimulatory factors (signal 1 and signal 2). Much more is known now about DC subsets and their ability to provide T cells with specific signals that differentially regulate the development of Th1 and Th2 responses control the state of immunity as well as tolerance. Our goals are 1) to understand what specific DC subsets do in vitro and in vivo to affect customized responses to distinct antigen, in particular tumor antigens;2) to understand how their in vivo function can be modulated in animal models;3) to test DC as therapeutic agents in the clinic. In three projects and two cores we are testing several hypotheses: hypothesis I is that DC regulate immune responses and determine the difference between a beneficial and a detrimental outcome;hypothesis II is that specific subsets of DC may be a useful vaccination tool to induce the immune responses with a desired character;hypothesis III is that the ability of DC subsets to control immunity and types of immune responses, depends on the expression of specific co-stimulatory molecules as well as the production of specific cytokines;hypothesis IV is that the form of antigen that is encountered by the DC is an important factor in the final outcome.

vaccine development; cyclins; mouth neoplasms; neoplasm /cancer vaccine; tumor antigens; neoplasm /cancer immunology; squamous cell carcinoma; recombinant proteins; cytotoxic T lymphocyte; nonhuman therapy evaluation; immunomodulators; neoplasm /cancer immunotherapy; helper T lymphocyte; patient oriented research; laboratory mouse; human subject;

DESCRIPTION (provided by applicant): We propose to design and test several MUC1 vaccines in preclinical studies in animals and in clinical trials in cancer patients. MUC1 is an epithelial tumor antigen. The vaccine design will be based on the new understanding of MUC1 immunogenicity derived through the aims of old Project 4 as well as the new knowledge about antigen uptake, presentation, and DC activation derived through the other projects in the PO1. We learned that the potential repertoire of MUC1 specific T cells could be much larger than had been observed in cancer patients. Difficulty we discovered in processing of the MUC1 glycoprotein by dendritic cells, severely limits the immune repertoire in vivo and, presumably the anti-tumor efficacy of anti-MUC1 immune responses. The hypothesis to be tested in pre-clinical studies in animals (Aim 1) and in clinical trials (Aim 2) is the following: if MUC1 glycoprotein is administered in forms that promote efficient processing and presentation by dendritic cells, resulting in a large number of peptide and glycopeptide epitopes, the anti-MUC1 T cell repertoire will broaden (including generation of helper T cells) and this will increase the tumor rejection potential of anti-MUC1 immune responses. The specific alms are: SPECIFIC AIM 1. Sub-aim a) To test in MUC1 transgenic mice the tumor rejection potential of cancer vaccines composed of a MUC1 peptide and/or glycopeptide that can be processed and presented by DC; Sub-aim b) To better understand the mechanisms of immune surveillance (and failure thereof) of MUC 1 + tumors, by examining the effect of uptake and processing of various forms of MUC1 on the biology of various DC populations. SPECIFIC AIM 2. To test in phase I/II clinical trials toxicity and immunogenicity of cancer vaccine, composed of a MUC1 peptide and/or glycopeptide processed and presented by DC and to evaluate immune effector mechanisms generated. The importance for tumor rejection and tumor specificity of soluble vs. particulate antigen and of peptide-specific versus glycopeptide-specific immune responses will be evaluated. The goal of both aims is to show that MUC1 is immunogenic in animals and patients and that the immune response it elicits could be a tumor rejection response.

DESCRIPTION (provided by applicant): Immunological solutions to cancer problem lie in our ability to elicit effective, potent immune responses, with life long memory, against antigens that are not much different then self molecules. The problem is compounded by different and often immunosuppressive effects on the immune cells by tumor cells. This exciting field of research continues to provide challenges and opportunities for young scientists well trained in the newest paradigms in immunology. The immunology faculty at the University of Pittsburgh School of Medicine have successfully trained many talented investigators in the past who have already made significant impact in the field of tumor immunology. In the first funding period of this grant, highly accomplished trainees were recruited and supported in their research related to tumor immunology. We propose to continue to recruit the best and the brightest into this area of research through the continuation of our predoctoral and postdoctoral training program in Cellular and Molecular Mechanisms of Tumor Rejection. We train predoctoral students and postdoctoral fellows within our Graduate Training Program in Immunology, one of nine training programs of the Interdisciplinary Training Program in Biomedical Sciences of the University of Pittsburgh School of Medicine. The goals supported by this training grant are to provide students and fellows with courses, research projects, highly qualified mentors and financial resources which will allow them to: 1. Elucidate the basic mechanisms of initiation of tumor-specific immunity; 2. Define the effector mechanisms in the anti-tumor immune response; and 3. Understand the requirements for establishment of long-term antitumor immune memory. We have been accomplishing these goals through well-planned and organized course activities, combined with program activities that promote substantive communication among the program faculty and trainees. The program also strives to train both graduate students and fellows in other important aspects of their profession: 1. Reporting in a timely fashion research results in peer reviewed manuscripts and at national and international meetings; 2. Writing grant proposals to various private and government funding agencies; 3. Teaching in courses related to their field of expertise.

antibody; neoplasm /cancer

DESCRIPTION (provided by applicant): We propose to design and test several MUC1 vaccines in preclinical studies in animals and in clinical trials in cancer patients. MUC1 is an epithelial tumor antigen. The vaccine design will be based on the new understanding of MUC1 immunogenicity derived through the aims of old Project 4 as well as the new knowledge about antigen uptake, presentation, and DC activation derived through the other projects in the PO1. We learned that the potential repertoire of MUC1 specific T cells could be much larger than had been observed in cancer patients. Difficulty we discovered in processing of the MUC1 glycoprotein by dendritic cells, severely limits the immune repertoire in vivo and, presumably the anti-tumor efficacy of anti-MUC1 immune responses. The hypothesis to be tested in pre-clinical studies in animals (Aim 1) and in clinical trials (Aim 2) is the following: if MUC1 glycoprotein is administered in forms that promote efficient processing and presentation by dendritic cells, resulting in a large number of peptide and glycopeptide epitopes, the anti-MUC1 T cell repertoire will broaden (including generation of helper T cells) and this will increase the tumor rejection potential of anti-MUC1 immune responses. The specific alms are: SPECIFIC AIM 1. Sub-aim a) To test in MUC1 transgenic mice the tumor rejection potential of cancer vaccines composed of a MUC1 peptide and/or glycopeptide that can be processed and presented by DC; Sub-aim b) To better understand the mechanisms of immune surveillance (and failure thereof) of MUC 1 + tumors, by examining the effect of uptake and processing of various forms of MUC1 on the biology of various DC populations. SPECIFIC AIM 2. To test in phase I/II clinical trials toxicity and immunogenicity of cancer vaccine, composed of a MUC1 peptide and/or glycopeptide processed and presented by DC and to evaluate immune effector mechanisms generated. The importance for tumor rejection and tumor specificity of soluble vs. particulate antigen and of peptide-specific versus glycopeptide-specific immune responses will be evaluated. The goal of both aims is to show that MUC1 is immunogenic in animals and patients and that the immune response it elicits could be a tumor rejection response.

Aberrant expression of cell cycle regulatory proteins characterizes many human tumors, including lung cancer. Dysregulated expression of cyclin B1 (CB1), demonstrated by constitutive overexpression and mislocalization in the cytoplasm rather than the nucleus, is especially significant because it contributes to malignant transformation and increased metastatic potential of tumor cells. Aberrant expression of CB1 is a result of functional inactivation of the tumor suppressor gene p53, an early event in the progression from premalignant to malignant lesions in the lung, suggesting that it could be a marker and a potential target in early as well as late stage cancer. CB1 overexprression in lung cancer elicits specific antibodies and T cells. CB1-specific antibodies are also seen in heavy smokers who are at high risk for developing lung cancer. Experiments, in mice show that anti-CB1 specific immune responses can protect from tumor challenge. The first hypothesis to be tested in Project 2 is that vaccines will elicit or boost CB1-specific immunity in lung cancer patients and that will result in an anti-tumor effect. Our second hypothesis is that the immune response against CB1 could be a biomarker of risk for development of future lung cancer in subjects with a positive smoking history or for recurrence among early-stage patients newly diagnosed with lung cancer. These two hypotheses are being tested in three specific aims. In Specific Aim 1 we propose to carry out clinical trials testing CB1 vaccines safety and immunogenicity in stage I and II lung cancer patients undergoing tumor resection. In Specific Aim 2 we will analyze and compare the quantitative and qualitative features of anti-cyclin B1 immunity in cancer patients and high-risk individuals in an attempt to elucidate important immune correlates of protection. In Specific Aim 3 we propose to evaluate the diagnostic and prognostic value of anti-CB1 antibodies in high risk individuals and cancer patients and to compare it to other diagnostic and prognostic markers studied in the SPORE projects. The overall goal is to translate the new knowledge acquired in the past grant period on the mechanisms of CB1 dysregulation and anti-CB1 immunity, to diagnosis, prognosis and therapy of lung cancer.

DESCRIPTION (provided by applicant): In the United States today there are as many 50-year olds as 5-year olds, and in 30 years there will be as many people over 80 as under 5 (www.census.gov/ipc/www/idbpvr.html). As the disease of old age, cancer will remain a great public health problem, if not become an epidemic. This extraordinary problem presents a great challenge to basic and applied biomedical research scientists to come up with extraordinary ideas to understand the disease and manage, cure or prevent it. The progress has been impressive in many scientific disciplines but a lot remains to be learned, discovered and applied. This work will be carried out by the new generation of biomedical scientists that we are now training in our laboratories. The overall goal of our training program is to train a cadre of young immunologists. PhDs and MD/PhDs. who will make it their mission to understand how to harness the immune system in the fight against cancer. Immunological solutions to cancer will lie in our ability to elicit effective, potent immune responses, with life long memory, against antigens that are not much different then self-molecules. This exciting field of research continues to provide challenges and opportunities for young scientists well trained in the newest paradigms in immunology. At the University of Pittsburgh School of Medicine we have successfully trained talented investigators who have made significant impact in the field of tumor immunology. We propose to continue to recruit the best and the brightest into cancer immunology through the continuation of our training program in Cellular and Molecular Mechanisms of Tumor Rejection. The goals supported by this training grant are to provide students and fellows with courses, research projects, highly qualified mentors and financial resources to: 1. Elucidate the basic mechanisms of initiation of tumor-specific immunity;2. Define the effector mechanisms in the anti-tumor immune response;and 3. Understand the requirements for establishment of long-term anti-tumor immune memory;4. Elucidate specific features of the tumor microenvironment that affect anti-tumor immunity;5. Elucidate the relationship between inflammation and cancer. We have been accomplishing these goals through well-planned and organized course activities, combined with program activities that promote substantive communication among the program faculty and trainees.

Seeinstructions): In the United Sates today there are as many 50-year olds as 5-year olds, and in 30 years there will be as many people over 80 as under 5 (www.census.gov/ipc/www/idbpvr.htmn. As the disease of old age, cancer will remain a great public health problem, if not become an epidemic. This extraordinary problem presents a great challenge to basic and applied biomedical research scientists to come up with extraordinary ideas to both understand the disease and manage, cure or prevent it. The progress has been impressive in many scientific disciplines but a lot remains to be learned, discovered and applied. This work will be carried out by the new generation of biomedical scientists that we are now training in our laboratories. The overall goal of our training program is to train a cadre of young immunologists. PhDs and MD/PhDs. who will make it their mission to understand how to harness the immune system in the fight against cancer. Immunological solutions to cancer will lie in our ability to elicit effective, potent immune responses, with life long memory, against antigens that are not much different then self-molecules. This exciting field of research continues to provide challenges and opportunities for young scientists well trained in the newest paradigms in immunology. At the University of Pittsburgh School of Medicine we have successfully trained talented investigators who have made significant impact in the field of tumor immunology. We propose to continue to recruit the best and the brightest into cancer immunology through the continuation of our training program in Cellular and Molecular Mechanisms of Tumor Rejection.The goals supported by this training grant are to provide students and fellows with courses, research projects, highly qualified mentors and financial resources to: 1. Elucidate the basic mechanisms of initiation of tumor-specific immunity; 2. Define the effector mechanisms in the anti-tumor immune response; and 3. Understand the requirements for establishment of long-term anti-tumor immune memory; 4. Elucidate specific features of the tumor microenvironment that affect anti-tumor immunity; 5. Elucidate the relationship between inflammation and cancer. We have been accomplishing these goals through well-planned and organized course activities, combined with program activities that promote substantive communication among the program faculty and trainees. RELEVANCE (See instructions):

Abstract My research over the last 25 years has been unique in providing data and scientific leadership on the path towards a prevention vaccine for non-viral cancers. Cancer immunotherapy is now a reality and both academia and pharmaceutical companies are focusing on the next best treatment for advanced cancer. This is unfortunate given the fact that even as successful as some cancer immunotherapy was recently shown to be, it helps only a fraction of patients and cures even a smaller subset of those. It is also the most expensive cancer treatment making it less likely to be broadly available to all who need it around the world. What is needed to focus attention of the field on immunoprevention are a few clinical successes and relevant basic science data to help interpret those and point a path forward. My research has been providing such data. Currently I am providing first examples of preventative cancer vaccines in clinical trials. With my clinical colleagues, FDA approval and NCI support, I completed the first in the world clinical trial testing a vaccine based on a human tumor antigen MUC1 in people at high risk for colon cancer This feasibility study enabled the second, currently ongoing, multi-center, double-blind, randomized, placebo controlled efficacy trial that will complete accrual in the Spring of 2016. In analyzing these trials we are obtaining new data, some of it very unexpected, important for the next set of preventative cancer vaccines that I am planning to develop, but also applicable to other vaccines and cancer immunotherapies. In the next seven years, my program will be focused on 1) Learning as much as possible from my first preventative vaccine trials; defining parameters controlling the response to and potential efficacy of the MUC1 vaccine through extensive molecular, cellular and proteomic analysis of PBMC and tissue samples; 2) Providing data in support of other premalignant lesions as candidates for preventative cancer vaccines and designing new cancer prevention trials; 3) Developing a universal vaccine for prevention and control of cancer as well as pathogens. Our work will hopefully encourage other cancer immunologists and immunotherapists to consider immunoprevention of non-viral cancers as an important approach to reducing the cancer epidemic.