Michael A. Caligiuri - US grants

Natural killer (NK) cells are operationally defined as cells capable of mediating spontaneous in vitro cytotoxicity against a variety of target cell populations without prior sensitization or MHC restriction. They express NKH1 while classic CTL, B cells, and myeloid cells do not. Their in vivo function, lineage, and site of maturation remain unknown. NK cells express CD2, and each can undergo activation through the CD2 pathway. The majority of peripheral blood CD2-CD3-NKH1-cells lack TCR gene rearrangement suggesting either a non-T cell lineage or a common precursor cell which is prethymic. NK cells are the first lymphocytes to return following T cell depleted bone marrow transplant, but the significance remains unclear. Freshly isolated human thymocytes lack expression of NKH1 and non-MHC- restricted cytotoxicity, but short term culture in rIL-2 results in the induction of NKH1 expression on NKH1- thymocytes and the development of NK killing. CD3-NKH1+ thymocytes can be generated from such cultures, but the precursor population is unknown. A subpopulation of early human thymocytes (CD2+CD3-) express IL-2R, produce IL-2 autocrine pathway, which may lead to proliferation and differentiation of functionally diverse cells. This early thymocyte may lead to proliferation and differentiation of functionally diverse cells. This early thymocyte may serve as a precursor for the IL-2 cultured CD3-NKH1+ NK cell and possibly for the CD3-NKH1+ NK cell found in human peripheral blood. Several possibilities may exist: 1) Within the thymus, the CD2+CD3- thymocyte is committed to expression of the TCR gene products. With continued exposure to IL-2 in vivo, some progeny differentiate into CD3+NKH1+ NK cells only, but no CD3-NKH1+ cells are thymic-derived in vivo 2) The CD2+CD3- thymocyte is not yet committed to express the TCR gene products, and with continued exposure to IL-2 in vivo, a fraction of thymocyte progeny differentiate into CD3\NKH1+ NK cells within human thymus and exit to the periphery. 3) A population of CD2+CD3- progenetor cells exist outside the thymus and its therefore not able to rearrange TCR genes. Through an IL-2 autocrine pathway, functional development of the CD3-NKH1+ cell would occur without TCR gene rearrangement or expression. Specific aims in phase I of this proposal involve an extensive in vitro phenotypic functional and molecular characterization of early thymocyte subpopulations, their progeny, and the CD3-NKH1+ cultured thymocytes. The data obtained should offer significant insights into the possible origin(s) and lineage(s) of human NK cells. Phase II will focus on further characterization of the NKH1 antigen and growth factors.

Natural killer (NK) cells are operationally defined as cells capable of mediating spontaneous in vitro cytotoxicity against a variety of target cell populations without prior sensitization or MHC restriction. They express NKH1 while classic CTL, B cells, and myeloid cells do not. Their in vivo function, lineage, and site of maturation remain unknown. NK cells express CD2, and each can undergo activation through the CD2 pathway. The majority of peripheral blood CD2-CD3-NKH1-cells lack TCR gene rearrangement suggesting either a non-T cell lineage or a common precursor cell which is prethymic. NK cells are the first lymphocytes to return following T cell depleted bone marrow transplant, but the significance remains unclear. Freshly isolated human thymocytes lack expression of NKH1 and non-MHC- restricted cytotoxicity, but short term culture in rIL-2 results in the induction of NKH1 expression on NKH1- thymocytes and the development of NK killing. CD3-NKH1+ thymocytes can be generated from such cultures, but the precursor population is unknown. A subpopulation of early human thymocytes (CD2+CD3-) express IL-2R, produce IL-2 autocrine pathway, which may lead to proliferation and differentiation of functionally diverse cells. This early thymocyte may lead to proliferation and differentiation of functionally diverse cells. This early thymocyte may serve as a precursor for the IL-2 cultured CD3-NKH1+ NK cell and possibly for the CD3-NKH1+ NK cell found in human peripheral blood. Several possibilities may exist: 1) Within the thymus, the CD2+CD3- thymocyte is committed to expression of the TCR gene products. With continued exposure to IL-2 in vivo, some progeny differentiate into CD3+NKH1+ NK cells only, but no CD3-NKH1+ cells are thymic-derived in vivo 2) The CD2+CD3- thymocyte is not yet committed to express the TCR gene products, and with continued exposure to IL-2 in vivo, a fraction of thymocyte progeny differentiate into CD3\NKH1+ NK cells within human thymus and exit to the periphery. 3) A population of CD2+CD3- progenetor cells exist outside the thymus and its therefore not able to rearrange TCR genes. Through an IL-2 autocrine pathway, functional development of the CD3-NKH1+ cell would occur without TCR gene rearrangement or expression. Specific aims in phase I of this proposal involve an extensive in vitro phenotypic functional and molecular characterization of early thymocyte subpopulations, their progeny, and the CD3-NKH1+ cultured thymocytes. The data obtained should offer significant insights into the possible origin(s) and lineage(s) of human NK cells. Phase II will focus on further characterization of the NKH1 antigen and growth factors.

The Center for HIV-Related Malignancies at Roswell Park Cancer Institute (RPCI) is the main treatment center for patients with AIDS-associated malignancies in Western New York. Under the direction of Dr. Zale Bernstein, the Center has had a rapid increase in patient referrals during the past two years. This, combined with Dr. Bernstein's commitment to protocol development and implementation has resulted in outstanding accrual of patients to both Institute and multi-institutional clinical trials, on par and in some instances exceeding accrual to protocols at more nationally prominent centers. In addition, two phase I clinical trials with biologic response modifiers have been completed within the past year. An extensive laboratory program of lymphocyte biology has been developed by Dr. Michael Caligiuri at RPCI that focuses on cytokine and cytokine receptor expression, as well as the characterization of human lymphomagenesis in the severe combined immune deficient mouse. This work has been translated into a number of Phase I and Phase II clinical trials, one of which has been completed by Dr. Bernstein in AIDS malignancy and is now being developed by the CALGB for a Phase II cooperative study. The Institute also has outstanding expertise in Flow Cytometry, a core facility that has been vital to the clinical and laboratory work of Drs. Bernstein and Caligiuri, respectively. With excellent patient accrual to clinical protocols in AIDS malignancies, strong data management and protocol support, and a continual source of laboratory based ideas for immune modulation using biologic modifiers for therapy of immune deficiency, RPCI has assembled a strong team of investigators who are likely to provide unique laboratory resources for correlative studies, substantial innovation in the design of clinical trials and patient accrual for rapid completion of such trials within the AIDS Malignancies Clinical Trials Consortium.

DESCRIPTION (provided by applicant): The Ohio State University (OSU) postdoctoral oncology training program is designed to produce physician-scientists and basic scientists capable of conducting independent research related to cancer treatment and prevention. In the 33-year history of this training grant at OSU, 139 postdoctoral students have been trained in basic cancer research from a cadre of outstanding NIH funded training faculty. Of these 139, 95 have had the MD degree, 35 the PhD degree, 6 with both MD/PhD degrees, and 3 with combined DVM/PhD degrees. Collectively 119 or 85% of these trainees have engaged in academic pursuits for all or a significant portion of their careers after leaving the fellowship program. In this last grant cycle, 32 of 37 T32 trainees (86%) are pursuing professional careers in cancer research or are still in professional training. Trainees will be selected from motivated and gifted M.D., D.O., DVM, or Ph.D. candidates who are committed to careers in patient-oriented cancer research. Trainees with health professional degrees will be selected from programs in medical, pediatric, surgical, and/or radiation oncology, and will have completed their clinical subspecialty training. To be selected, physician candidates will need to demonstrate an appreciation of the need for physician-scientists to link the bench with the bedside. Candidates possessing the Ph.D. degree will be chosen from basic science programs of biochemistry, molecular genetics, microbiology and immunology, pharmacology, virology, cellular biology, mathematics, physics, or other programs which have prepared and stimulated the candidates to translate their basic research toward the solution of clinical cancer research questions. The training program will require two to three years. Trainees will have independent research projects within a training faculty's laboratory. All training faculty are members of one of seven OSU Comprehensive Cancer Center (CCC) Programs of: Cancer Control, Experimental Therapeutics, Hormones and Cancer, Immunology, Molecular Biology and Cancer Genetics, Molecular Carcinogenesis and Chemoprevention, Oncogenic Virus. The 48 training faculty and the 13 contributing faculty are all NIH/NCI/DOD/ACS-funded basic scientists and physician-investigators who have developed a network of collaborative interactions within the CCC which expose the trainee to multiple research perspectives and expertise. The same faculty participates with trainees in joint conferences which further enhance appreciation of different research approaches. Formal courses assist the trainee in developing an appropriate knowledge base. This competitive renewal application seeks continued support for the 8 trainees each year, with half to a majority of those being physician-trainees.

DESCRIPTION: The applicants hypothesize that assessment of NK cell development and function in patients with HIV-1 infection will provide insights into how the human immune system can be better modulated to control HIV-1 replication and to improve the host response to infection. The specific aims of the proposal are: 1) to assess human NK cell differentiation from CD34+ hematopoietic bone marrow progenitor cells in patients with controlled and uncontrolled HIV-1 infection; 2) to assess the macrophage-NK innate immune effector arm in patients with controlled and uncontrolled HIV-1 infection following infectious insult in vitro; and 3) to assess NK cells obtained from patients with controlled and uncontrolled HIV-1 infection for their ability to suppress HIV-1 replication in vitro. To accomplish these objectives, the applicants propose to study 40 patients with HIV-1 infection stratified into 4 cohorts based on anti-retroviral medication, viral load and absolute CD4 cell count. They will also include 20 age-matched normal controls. Bone marrow stromal cells will be purified using methods previously developed in the applicant's laboratory and these cells will also be used to evaluate NK cell differentiation from purified CD34+ hematopoietic progenitor cells (HPC) in the presence of IL-15, KL and FL. In specific aim 2, a monocyte-NK cell co-culture system that has previously been developed will be used to assess the ability of monocytes to secrete monokines in response to various stimuli as well as the ability of NK cells to respond to endogenous monokine stimulation through the production of IFN-gamma. Specific aim 3 will examine the ability of supernatants from monokine-stimulated NK cells to suppress HIV-1 replication in vitro. These studies will examine both M-tropic and T-tropic strains of HIV.

The median time of survival for patients with the acquired immune deficiency syndrome diagnosed with primary central nervous system lymphoma (AIDS PCNSL) is less than four months, despite radio- and chemotherapy and recent improvements in the suppression of HIV-1 replication. Essentially no advances have been made in the clinical management of this disorder since its initial characterization in 1991 as a lymphoma associated with the Epstein-Barr virus (EBV) in virtually 100 percent of cases. Death in patients with AIDS PCNSL results as often from therapy-induced toxicity and complications of AIDS as it does from progression of the PCNSL. To the best of our knowledge, an in vivo preclinical animal model to explore novel strategies for the treatment of AIDS PCNSL has not been described. The current proposal is a revised application. In our initial proposal, there was considerable enthusiasm for our discovery and characterization of Fas expression on AIDS PCNSL in the absence of Fas expression in human brain parenchyma, and our single specific aim to develop an animal model of PCNSL to explore anti-Fas therapy. Over the last year our laboratory has worked to successfully develop a model of human PCNSL in the nude rat through a multidisciplinary collaboration involving several investigators at our university. The model utilizes human EBV+ B cell tumors that arise spontaneously in the setting of immune deficiency to implant into the CNS of the nude NIH rat. Within 13 days of implantation, the rat develops irreversible neurologic symptoms from an expanding tumor mass within the CNS. Histologic evaluation at autopsy reveals an infiltrating human EBV+ B cell tumor. Magnetic resonance imaging is used to document progression in tumor growth and provide accurate assessment of tumor volume. Commercial instrumentation allows reproducible implantation and therapeutic delivery into the rat CNS that is comparable to delivery of CNS therapies in humans. The first specific aim of the revised proposal will focus on refinement of the preclinical animal model to best assess toxicity and efficacy of novel antitumor strategies in vivo. We hypothesize that novel therapies which target EBV+ lymphoma cells in vitro can be meaningfully explored in a preclinical in vivo animal model of PCNSL prior to early clinical development in patients with AIDS PCNSL. The model should, therefore, assist in determining which in vitro observations are too toxic or simply ineffective to pursue in vivo, and provide support for the transition of promising basic observations to clinical trials for the treatment of AIDS PCNSL. In the second and third specific aims of this revised proposal, two such in vitro observations will be pursed in the in vivo animal model: 1) a combination of antiviral therapy and cytokine therapy that induces apoptosis of EBV+ cell lines in vitro and has shown promising results in a few patients with AIDS PCNSL in vivo, and 2) anti-Fas therapy that induces apoptosis of AIDS PCNSL in vitro. The limitations of the animal model preclude its use for studying the pathogenesis of AIDS PCNSL.

This application is a five year competing renewal for the AIDS Malignancy Consortium (AMC) grant currently awarded to Michael A. Caligiuri, M.D. at The Ohio State University (OSU). During the past four years of this award, the PI was an active participant in the AMC Lymphoma Working Group and the AMC Laboratory Working Group. The PI successfully competed for correlative science awards for two AMC clinical trials, and the PI currently chairs one AMC clinical protocol that uses a biologic response modifier in HIV non Hodgkin's lymphoma (NHL). Two additional clinical studies are currently under development by the PI for the AMC: The first is a randomized trial of low dose interleukin (IL) 2 following first induction therapy in HIV NHL. This study will likely be performed in collaboration with industry and AIDS malignancy sites in Europe. The letter of intent (LOI) was reviewed by the AMC and a protocol has been submitted to the AMC. The second study submitted by the PI is a phase II study assessing the anti-tumor activity of anti-CD20 monoclonal antibody against patients with posttransplant lymphoproliferative disorder (PTLD). PTLD will now be incorporated into the AMC agenda as an immunodeficiency lymphoma, and this will be the first such protocol within the AMC. The LOI as been approved by the AMC and the protocol has been submitted. Despite these intellectual contributions, the accrual of OSU and its former affiliate, Roswell Park Cancer Institute, was poor, ranking approximately 8th among 13 primary AMC sites. Therefore, in order to address this weakness, the PI has now affiliated with four new sites, each with a high patient volume of HIV-1+ patients and patients with AIDS malignancies, and each a new member to the AMC. The PI is no longer affiliating with Roswell Park. These four new sites include the University of Maryland Cancer Center, The Brady Memorial Hospital of Emory University, Saint Vincent's Comprehensive Cancer Center in New York, and a consortium of three hospitals in Australia that function under a common clinical research group called the National Center for HIV Epidemiology and Clinical Research (NCHECR). The NCHECR evaluates and treats the vast majority of HIV-1 and AIDS malignancy patients for all of Australia. Each of the OSU- affiliated sites has unique strengths. Some centers have large inner city populations with high volumes of women and minority patients, while other centers have extremely strong histories of phase I-III cooperative group trials or unique laboratory expertise. Collectively, this new group of OSU-affiliated sites should bring several strengths to the AMC, most notably an increase in accrual to AMC protocols for HIV NHL and HIV Kaposi's sarcoma. A budget has been structured to provide a minimal baseline of support for each affiliated site to get protocols approved by Institutional Review Boards and to begin to screen patients for study. However, after an initial accrual of four patients per site, the funding of each site becomes tied to their ability to accrue patients. Collectively, this application provides enhanced strength in intellectual contributions and patient accrual for the AMC, compared to our previous application.

The Ohio State University (OSU) postdoctoral oncology training program is designed to produce physician-scientists and basic scientists capable of conducting independent research related to cancer treatment and prevention. Trainees will be selected from motivated and gifted M.D., D.O., or Ph.D. candidates who are committed to careers in patient- oriented cancer research. Trainees with health professional degrees will be selected from programs in medical, pediatric, surgical, and/or radiation oncology, and will have completed their clinical subspecialty training. To be selected, physician candidates will need to demonstrate an appreciation of the need for physician-scientists to link the bench with the bedside. Candidates possessing the Ph.D. degree will be chosen from basic science programs of biochemistry, molecular genetics, microbiology and immunology, pharmacology, virology, cellular biology, mathematics, physics, or other programs which have prepared and stimulated the candidates to translate their basic research toward the solution of clinical cancer research questions. The training program will require to two to three years. Trainees will have independent research projects within a training faculty laboratory. All training faculty are members of one of eight Comprehensive Cancer Center (CCC) Programs of: molecular biology and cancer genetics, immunology, developmental therapeutics, hormones and cancer, molecular carcinogenesis, cancer prevention and control, RNA tumor viruses, or neuro-oncology. The 46 training faculty and the 12 contributing faculty are all NIH/ACS-funded basic scientists and physician-investigators who have developed a network of collaborative interactions within the CCC which expose the trainee to multiple research perspectives and expertise The same faculty participate with trainees in joint conferences which further enhance appreciation of different research approaches. Formal courses assist the trainee in developing an appropriate knowledge base. The new Division of Human Cancer Genetics and revitalized Division of Hematology-Oncology within the OSU CC has resulted in 18 new faculty recruitments during the past academic year, with 10 more faculty positions in these two Divisions to be filled in the current academic year, all with a cancer focus. This competitive renewal application seeks support for 10 trainees by the end of the fifth year of funding with 7 or 8 of those being physician-trainees.

interleukin 15; immunoregulation; cellular immunity; natural killer cells; leukocyte activation /transformation; hematopoietic stem cells; biological signal transduction; clinical research; polymerase chain reaction; southern blotting; genetically modified animals; laboratory mouse; human subject; tissue /cell culture;

DESCRIPTION (Provided by applicant): Patients with the acquired immune deficiency syndrome (AIDS) are highly susceptible to the development of fatal infection and cancer. If the durability of responses to highly active anti-retroviral therapy (HAART) is limited, then alternative or complementary therapeutic measures directed toward the preservation of immune competence will need to be considered to prevent fatal infection and/or malignant transformation. In the case of preventing systemic HIV lymphoma, such strategies appear necessary even with effective HAART. We have designed a novel strategy to modulate a component of the innate immune system (natural killer or NK cells) and to modulate the cytokine gene profile in patients with AIDS and AIDS malignancies. We believe this trial to be the first of its kind in the world, and it is based largely on studies performed in our laboratory and our clinic over the last seven years. We identified co-expression of the receptor tyrosine kinase c-kit and the interleukin (IL)-2 receptor on lymphocytes, and demonstrated functional synergy following the binding of their respective ligands, stem cell factor (SCF) and IL-2. We demonstrated the synergy of SCF plus low dose IL-2 in inducing expansion of human NK cells in vitro from CD34+ hematopoietic stem cells, compared to either SCF or IL-2 alone, followed by a similar demonstration in an in vivo animal model. We performed studies of low dose IL-2 alone in patients with AIDS and AIDS malignancies, along with correlative laboratory studies. We are now poised to assess the safety, feasibility, and immunomodulatory effects of administering a combination of SCF and IL-2 to patients with AIDS and AIDS malignancy. This application seeks support to perform correlative laboratory studies to assess the cytokine gene profile in vivo, and the phenotype and function of hematopoietic stem cells, T cells, and NK cells before, during and after patients receive IL-2 alone or SCF and IL-2 in a Phase I IRB-approved outpatient study. The overall objective of this application is to characterize the immunologic consequences of this innate immune modulation to better understand how to compensate for antigen-specific immune deficiency.

DESCRIPTION (Provided by Applicant): In this R13 proposal for NIH support of a single scientific meeting, we will present a detailed outline for the 19th Annual International Natural Killer Cell Workshop/7th Annual Meeting of the Society for Natural Immunity to be held October 7-9, 2001 at The Caribe Hilton Hotel in San Juan, Puerto Rico, a Commonwealth Island of the United States of America. The participants will consist of M.D., Ph.D., and M.D.-Ph.D. basic and clinical scientists interested in innate immunity, natural killer cell biology, and clinical applications of natural killer cell biology. The meeting is generally attended by 150-250 participants that include virtually all the leaders in the field of natural killer cell biology as well as pre-doctoral and post-doctoral students that comprise approximately 50 percent of the attendees. The goals of the meeting are to: 1) bring together senior and junior level researchers in the field of NK cell biology, and also those in related areas of non- adaptive immunity; 2) provide a forum for presenting and discussing the newest and most significant developments in select areas of non-adaptive immunity and T/NK leukemia; 3) provide a special opportunity for a select group of students and faculty of Hispanic origin from Puerto Rico to participate in a pre- meeting mini-course on NK cell biology and to participate in the meeting itself; 4) determine future directions in the field both in terms of basic research and clinical applications. The topics include: NK Differentiation, Nk/NKT Regulation, Clinical and Basic Aspects of NK Biology, NK Cell Signaling, NK Cell Receptor Expression and Function, and three workshops led by the leaders in the field of NK cell biology. Finally, there will be a distinguished lecture, delivered by Dr. Thomas Waldmann of the NIH on T-NK Cell Leukemia, reviewing the basic and clinical aspects. Through a formal collaboration with the University of Puerto Rico, there will be a series of pre-meeting lectures at the University on innate immunity by the 3 program organizers, and a concerted effort to bring minority Hispanic pre-doctoral and post-doctoral students to the meeting and to the U.S. for post-doctoral training. Funds are requested to support the latter effort as well as support for a fraction of the invited speakers and program planner.

DESCRIPTION (provided by applicant): This Program Project Grant (PPG) application stresses innovative clinical cancer immunotherapy trials based on our current understanding of innate immunology, as well as basic investigation into innate immune effector cell function in preparation for subsequent, more refined clinical cancer immunotherapy trials. Project 1 is focused on monoclonal antibody therapy for the treatment of non-Hodgkin.s lymphoma (NHL). In preclinical studies performed in the SCID-human chimeric mouse model of malignant human lymphoproliferation, the combined administration of rituximab plus interleukin (IL)-2 is curative, while the administration of either agent alone is ineffective. Mechanistic studies in the model are underway. The first clinical study will be a combination of rituximab + IL-2 for the treatment of relapsed NHL. A second clinical trial will examine pro-inflammatory cytokine blockade as a means to enhance efficacy and decrease toxicity associated with agents like rituximab and Campath. Additionally it will focus on clinical trials that are derived from basic studies in natural killer (NK) cell biology. Specifically, we will be investigating the co-administration of IL-2 and Flt3 ligand for their synergy in expanding NK cells in-patients with HIV and HIV associated malignancy. Projects 2-4 each investigate one aspect of either monocyte/macrophage or NK cell biology with the notion that improved understanding of innate immune effector cell function will contribute to the design and implementation of the subsequent set of immunotherapy trials. Project 2 is focused on Fc?R signaling in monocytes with broad application to NK cell biology; Project 3 investigates the regulation of activation and tumor cell recognition within two newly identified subsets of human NK cells; Project 4 uses murine models to assess the role of co-stimulatory molecules in NK cell recognition of tumor targets and characterize NK subsets similar to those found in humans. The PPG is supported by 4 cores: Administration, Biostatistics, Correlative Science and Mouse Facilities. The overall goal is to rapidly introduce innovative clinical trials testing laboratory-based hypotheses, while pursuing additional basic investigation of innate immunity for subsequent cancer immunotherapy trials.

DESCRIPTION: (provided by applicant) This is a revised application. All changes are indicated by, a red line in the right margin. At the molecular level, acute myeloid leukemia (AML) is a heterogeneous disease. Recent advances with molecular-based risk stratification of AML and molecular-based therapeutics strongly suggest that elucidation of molecular mechanisms underlying each case of AML will have the greatest impact on increasing the cure rate of this disease. In the majority of AML cases, cytogenetics are either normal or only contain changes in chromosome number that limit one's ability to find leukemogenic gene fusions. Several years ago, our laboratory collaborated to discover a novel molecular defect found in 5-10 percent of AML cases with normal cytogenetics and in the majority of AML cases with trisomy 11 as a sole abnormality. The defect involves a partial tandem duplication (PTD) of the MLL gene, whereby exons 2-6 or 2-8 duplicate in tandem creating a unique self-fusions. Our laboratory has since performed an extensive characterization of the MLL PTD. We hypothesize that the MLL PTD represents a primary molecular defect in myeloid hematopoietic progenitor cells that is responsible, at least in part, for their leukemic transformation. We propose a series of in vitro and in vivo model systems to test this hypothesis and to define the genetic differences that specifically result from the MLL PTD. We have developed a targeting construct to create embryonic stem cells expressing the MLL PTD for in vitro differentiation studies as well as for creation of chimeric and heterozygous mice expressing the MLL PTD. Finally, we have utilized two techniques to study genome-wide genetic and epigenetic changes in leukemic tissue to better understand downstream effector molecules during malignant cell growth, and propose to use these technologies to better understand pathways critical to leukemogenesis in cells harboring the MLL PTD. Ultimately, we believe insights gained by the experiments proposed in this application will further our understanding of leukemogenesis and open up new therapeutic options for this subset of AML patients with a poor prognosis.

The Cancer and Leukemia Group B (CALGB) Leukemia Correlative Sciences Program has worked to attain a highly successful integration of correlative laboratory studies into the design and implementation of CALGB leukemia trials. Dr. Michael A. Caligiuri assumed leadership of the CALGB Leukemia Correlative Science Committee (LCSC) in August of 1999 following 18 years of Committee leadership by Dr. Clara Bloomfield. The LCSC has been restructured and refocused entirely on leukemia, moving evermore into the molecular age of diagnostics, pathogenesis, and treatment. The goal is to use cytogenetics, immunology, and molecular biology to better understand the heterogeneity of leukemia with regard to diagnosis, prognosis, and ultimately treatment. In this competitive renewal application, we have assembled three Cores (Cytogenetics, Banking, and Administration) and six Projects, each of which is utilizing materials collected from patients treated on CALGB leukemia treatment protocols in order to address a scientific question and correlate it with disease outcome. Having set into motion a paradigm for stratification of leukemia treatment based on risk of relapse during this last funding cycle, the current CALGB Leukemia Correlative Science application will attempt to dissect out additional leukemia patients that can be predicted to do well or do poorly in response to standard or novel therapies, and to then work with the CALGB Leukemia disease committee to stratify their treatment based on risk. In this way, we hope to significantly impact on the cure rate of leukemia over the next six years of funding.

The Ohio State University Comprehensive Cancer Center (OSUCCC) is currently in its 24th year as an NCI-designated CCC and is now requesting continued federal support for the next five years. The OSUCCC came under its 3rd Director in August 1997 and has initiated a series of changes aimed at creating new scientific direction and revitalizing existing clinical disciplines. The overall goal remains to reduce cancer morbidity and mortality through continu4ed basic, translational and clinical research. To accomplish this the OSUCCC has, in the last 18 months, 1) created a new Division of Human Cancer Genetics with committed institutional support currently in excess of $50 million; 2) created, eliminated and restructured existing CCC programs to be consistent with existing scientific CCC programs to be consistent with existing scientific and clinical strengths and future direction; 3) recruited 32 programs to be consistent with existing scientific and clinical strengths and future direction; 3) recruited 32 new full time faculty to OSU, all with cancer relevant research and/or clinical care, including 9 physician scientists to medical or surgical oncology; 4) developed a new clinical trials infrastructu7re to better facilitate innovative clinical cancer research; 5) acquired a 64% increase in new space allocated for cancer research. The 201 OSUCCC members are currently served by 8 shared resources and are distributed among 7 programs: Cancer Prevention & Control, Experimental Therapeutics, Hormones and Cancer, Immunology, Molecular Biology & Cancer Genetics, Molecular and Cellular Carcinogenesis, and RNA Oncogenic Virus. These programs are supported by over $29 million in annual direct costs from NCI approved peer-reviewed funding, with $10.6 million of that funding coming from 95 NCI-sponsored projects, representing a 153% increase in NCI funding since last submission. The new OSUCCC director and her recruits have resulted in a substantial increase in University and philanthropic support, NCI-approved grant dollars. OSUCCC laboratory and clinical space, and published evidence of intra- and inter-programmatic collaboration. The changes in OSUCCC leadership, support for new scientific directions, and revitalization of translational and clinical cancer research efforts should greatly strengthen the OSUCCC's ability to achieve its overall goal during the next 5 years.

immune response; subcutaneous drug administration; interleukin 2; cellular immunity; antiviral agents; dosage; drug administration rate /duration; leukocyte activation /transformation; clinical trial phase I; hematopoietic stem cells; natural killer cells; drug adverse effect; virus load; Epstein Barr virus; cytotoxic T lymphocyte; combination therapy; clinical research; human subject;

Developmental Funds are critical to the future progress of the OSUCCC. During the prior funding period, the OSUCCC benefited from $1,936,630 in CCSG developmental funds. For the $485,000 of CCSG dollars used for faculty recruitment, the return to-date in total direct costs for CCSG-approved peer-reviewed extramural funding is $ $45,962,763, a 95:1 return on investment, and in total funding was $48,888,138, a 100:1 return on investment (Table 1). In addition, we used CCSG funds to establish three shared resources, the Pharmacoanalytical Shared Resource, the Biomedical Informatics Shared Resource and the Small Animal Imaging Shared Resource. A brief summary ofthe progress with these three facilities is described below. Their operations as fully established shared resources are described in Section 9.0 of this application.

DESCRIPTION (provided by applicant): The Ohio State University Comprehensive Cancer Center (OSUCCC) is currently in its 29th year as an NCI-designated CCC and is now requesting continued federal support for the next five years. The OSUCCC named its 4th Director, Michael A. Caligiuri, M.D. in July, 2003 while retaining the former Center Director, Clara D. Bloomfield, M.D., as a Senior Advisor to the OSUCCC. The overall goal remains to reduce cancer morbidity and mortality through continued basic, translational and clinical research. The 183 OSUCCC members are currently served by 13 shared resources and are distributed among 6 Research Programs: Cancer Control, Experimental Therapeutics, Immunology, Molecular Biology and Cancer Genetics, Molecular Carcinogenesis and Chemoprevention and Viral Oncogenesis. Since the last competitive renewal, the OSUCCC has shown significant growth as demonstrated by: 1) the recruitment of more than 90 new cancer research faculty to OSU;2) more than a 230% increase in patient accrual to investigator-initiated trials;3) the addition of 6 new shared resources at an institutional investment of over $10.8 M;4) a 144% increase in total NCI funding and a 65% increase in peer-reviewed funding. The OSUCCC has also seen tremendous growth in institutional commitment since 1999 as demonstrated by: 1) a new formal role for the OSUCCC Director within OSU, providing complete oversight of the University-wide cancer funding initiatives, opportunities and cancer grant submissions;2) A seat on the eight-member Medical Center Executive Committee which meets every week with the Senior VP of Health Sciences to plan and evaluate the entire Health System's direction (cancer is the only discipline represented on this Committee);3) a written commitment for the expansion of the Cancer Program facilities that will result in more than a four-fold increase over the current square footage under control of the OSUCCC Director at a cost of approximately $350 M;4) interim laboratory and office space for recruitment until the new facility is constructed;5) an additional $7.0 M of cash annually from OSU for research and infrastructure expansion;and 6) twenty senior faculty slots under the control of the OSUCCC Director, each provided with $100,000 of guaranteed salary in perpetuity. With these new resource commitments in place, the OSUCCC is poised for significant growth and expansion in the next 5 years.

In 2003 the OSUGCC Senior Leadership Team consisted of Michael A. Caligiuri, M.D. as Director, David E. Schuller, M.D. as Deputy Director, Michael Lairmore, D.V.M, Ph.D. as Associate Director for Basic Research, William E. Carson, M.D. as Associate Director for Clinical Research, Electra Paskett, Ph.D. as Associate Director for Population Sciences and Mr. Jeff Walker as Associate Director for Administration. At that time, the NCI review committee rated Senior Leadership as

? DESCRIPTION (provided by applicant): This proposal is now in its fourth cycle of competitive funding and focuses on Interleukin 15 (IL-15) and its role in human natural killer (NK) cell development, NK cell effector function and cancer. Our specific aims remain relatively unchanged: 1) To further identify and characterize progenitors and precursors that differentiate into human NK cells; 2) To better understand the role of IL-15 in regulating human NK cell effector functions. IL-15 induced differentiation and activation of NK cells are, like other biologc systems balanced by negative regulators, and those negative regulators are in turn exploited by tumors to turn off and evade the NK cell's ability to effectively survey against malignant transformation. Thus, our overall goal in this proposal is to gain a comprehensive understanding of the cellular and molecular components that constitute these inhibitory pathways in order to best understand how they work and how they can be modified to enhance NK cell development and function in the face of tumor challenge. In particular, in Aim 1 we will further investigate th innate cellular heterogeneity within secondary lymphoid tissues where we previously discovered and characterized human NK cell developmental intermediates. Over the past five years, an abundance of additional innate lymphoid cells (ILC) have been discovered and characterized, lending some controversy as to the origin of human NK cells. We hypothesize that the transcription factor aryl hydrocarbon receptor (AHR) acts to prevent human NK cell development from an IL-22 secreting ILC, and does so via the upregulation of miR-29b. Indeed, certain tumors have been shown to secrete AHR agonists that we postulate inhibit NK cell development. We propose a series of experiments to test our hypothesis, which if validated, will provide a new target for combating immune evasion. In Aim 2 we have preliminary data that suggest the activated Axl/Mer/Tyro3 receptor tyrosine kinase (RTK) pathway serves to negatively regulate IL-15-induced NK cell activation. Further, we have shown that the ligand to this RTK family, Gas6, is expressed on human acute myeloid leukemia cells and confers a poor prognosis. Thus, we hypothesize that this family of RTKs plays an important role in the negative regulation of NK cell effector functions and thus has the potential to be targeted for immune checkpoint inhibition in order to enhance NK cell-mediated immune therapy against cancer.

PROTOCOL-SPECIFIC RESEARCH SUPPORT In the past year (12/1/2002-11/30/2003), 139 patients were registered on 15 innovative, early-phase investigator-initiated therapeutic clinical trials reflecting the efforts of 8 different OCUCCC principal investigators (Table 2). These therapeutic protocols reflect efforts in four OSUCCC programs;Experimental Therapeutics (ETP), Cancer Control (CC), Molecular Carcinogenesis and Chemoprevention (MCC) and Immunology (IMM). These protocols and patients are managed by the Clinical Trials Office (CTO) utilizing protocol-specific-research nurse support. Based upon the current accrual statistics for investigator-initiated therapeutic clinical trials and an estimate of approximately 35 active participants per clinical research associate or research nurse per year, we can justify a request for 2.4 FTEs for protocol-specific research nurse support in year one of our CCSG renewal (see justification above). The OSUCCC CTO manages 318 trials, including 55 therapeutic trials that were written by local investigators. A number of these (15) are supported though the OSU phase 1 contract (U01 CA76576-03) and 9 others are funded via Quick Trial NIH R21 grants or separate R01s or P01s hence the seemingly low number of innovative phase I studies covered by the Protocol-Specific Research Mechanism is based not on a lack of innovation but rather on innovation rewarded with peer-reviewed funding for clinical research. In addition to those funded through R21s or R01s, investigators have been active in obtaining partial or full support for research efforts through industry partners. The investigators at the OSUCCC are encouraged to actively pursue funding for trials through external peer review mechanisms whenever possible.

The broad, long-term objective of this proposal is to understand how cancer evades and disarms the mmune system at the molecular, cellular and organism level so this process can be interrupted for the successful prevention of primary or recurrent cancer. This P01 competitive renewal application primarily investigates the human innate immune system and Project 3, while integrated with the efforts of Projects 1, 2 and 4, is focused on natural killer (NK) cells. Work with cancer patients receiving T-cell depleted HLA haplo- identical inhibitory killer immunoglobulin like receptor (KIR)-mismatched allogeneic stem cell transplant provides direct evidence that NK cells have a graft versus leukemia effect that correlates with improved survival. Understanding the molecules that regulate NK cell development and function at the molecular, cellular and organism level will be critical to effectively manipulate the immune system for the prevention and treatment of cancer. Two of the many molecules that appear important in NK cell responsiveness to the development of cancer are transforming growth factor beta (TGF-(3) and inhibitory KIRs. Specifically, this proposal will: 1) Investigate the basic mechanisms by which TGF-p exerts its effects on NK cell development and NK cell function. For these studies we will use fresh secondary lymphoid tissue (i.e., tonsils and lymph nodes) where human NK cells develop, along with fresh human blood as a source of primary mature NK cells. 2) Perform two clinical trials in which patients with melanoma, renal carcinoma and pancreatic cancer will receive a "first-in-man" therapy of neutralizing antibody against TGF-p. 3) Perform a third clinical trial in which patients with multiple myeloma will receive an antibody that has broad recognition of inhibitory KIR. In each of these three trials, we will use patients with existing cancer who have failed conventional therapy, and thus we will have the opportunity, with our P01 colleagues, to assess human NK cell development and function in vivo before, during and after these novel and potentially immune modulating therapies are delivered. Collectively, this work will synergize with Cores A, B, and C and with Projects 1, 2, and 4 of this P01 competitive renewal application to better understand human innate immune development and function as it occurs normally and in cancer patients. It will provide new insights as to how innate immune effector cells may be modulated in order to prevent cancer or its recurrence.

The Ohio State University Comprehensive Cancer Center (OSUCCC) is currently in its 29t"year as an NCI-designated CCC and is now requesting continued federal support for the next five years. The OSUCCC named its 4t" Director, Michael A. Caligiuri, M.D. in July, 2003 while retaining the former Center Director, Clara D. Bloomfield, MD, as a Senior Advisor to the OSUCCC. The overall goal remains to reduce cancer morbidity and mortality through continued basic, translational and clinical research. The 183 OSUCCC members are currently served by 13 shared resources and are distributed among 6 Research Programs: Cancer Control, Experimental Therapeutics, Immunology, Molecular Biology and Cancer Genetics, Molecular Carcinogenesis and Chemoprevention and Viral Oncogenesis. Since the last competitive renewal, the OSUCCC has shown significant growth as demonstrated by: 1) the recruitment of more than 90 new cancer research faculty to OSU; 2) more than a 230% increase in patient accrual to investigator-initiated trials; 3) the addition of 6 new shared resources at an institutional investment of over $10.8 M; 4) a 144% increase in total NCI funding and a 65% increase in peer-reviewed funding. The OSUCCC has also seen tremendous growth in institutional commitment since 1999 as demonstrated by: 1) a new formal role for the OSUCCC Director within OSU, providing complete oversight of the University-wide cancer funding initiatives, opportunities and cancer grant submissions; 2) A seat on the eight-member Medical Center Executive Committee which meets every week with the Senior VP of Health Sciences to plan and evaluate the entire Health System's direction (cancer is the only discipline represented on this Committee); 3) a written commitment for the expansion of the Cancer Program facilities that will result in more than a four-fold increase over the current square footage under control of the OSUCCC Director at a cost of approximately $350 M; 4) interim laboratory and office space for recruitment until the new facility is constructed; 5) an additional $7.0 M of cash annually from OSU for research and infrastructure expansion; and 6) Twenty senior faculty slots under the control of the OSUCCC Director, each provided with $100,000 of guaranteed salary in perpetuity. With these new resource commitments in place, the OSUCCC is poised for significant growth and expansion in the next 5 years.

The Administrative Core will provide centralized grant administration, communication processing, and budget management. This Core is critical in order to amalgamate the efforts of the Project investigators, their experimental findings and their ideas, to evaluate research efforts and to direct the summary efforts toward the Program outcome. In addition, in conjunction with the Biostatistics and Data Management Core (Core B), Core A will create and operate a secure web site for the interchange of data, communications, and the evaluation of Program progress. Core A will also provide the programmatic infrastructure to integrate the special services of Cores C, D, & E with the projects, and efficiently manage these resources. The Specific Aims of the Administrative Core are to: 1. Provide basic administrative services to the investigators. [unreadable] This includes the management of project supplies, filing, development of memos, meeting minutes and communications covering all operations, including publications. By design, this core will provide investigators with clear lines of scientific and administrative communication to promote collaboration among team members, aid in the prioritization of resources, and facilitate resolution of any problems that affect team members. [unreadable] To install data prioritization evaluation and inter-institutional secure website for sharing and storing data bases. Provide access in real time to data sets and standardization of data sets to permit data managers from all institutions to have input on final statistics and direct modeling and experimental design over the program course. 2. Organize monthly meetings of the Program Steering Committee. [unreadable] This Steering committee shall consist of the Project and Core Pis. 3. Organize an annual Internal Advisor Review meeting where a panel of experts shall assess the Program effectiveness and experimental progress, research directions, technical approaches, statistical evaluation, and administrative effectiveness. 4. Organize an annual External Review Panel meeting. [unreadable] The members of External Review Panel shall review all experimental findings and clinical outcomes, help prioritize investigations, review the coordination of collaborations, and evaluate concepts that emerge from the studies. 5. Provide publication services to the investigators for project-related communications where this includes the preparation of manuscripts, abstracts, publications, data sharing, IP, protocols, corporate agreements, and compliance issues. 6. Provide overall fiscal review, accounting, and real time budget analyses. [unreadable] To conduct and direct the budget analysis including both descriptive summary expenditures and real- time tracking of individual budgets. [unreadable] Provide forward-looking projections on expenditures. These can be correlated with project progress to provide superior Program management. [unreadable] Provide timely review, quality assurance, and dissemination of the budgets in all projects.

DESCRIPTION (provided by applicant): It is now clear that treatment of certain solid and liquid tumors with targeted monoclonal antibodies (mAb) can improve clinical outcome for many patients. It is also clear that in many instances this mAb therapy requires participation from the innate immune system. Nonetheless, tumors themselves exploit innate immune tolerance so as to weaken effective participation by the latter, thereby contributing to the fact that mAb therapy is still not curable today. In this competing renewal application we hypothesize that a better understanding of how the tumor exploits innate immune effector cell tolerance will lead to novel therapeutic options that largely reverse this process and enhance mAb therapy of cancer. To achieve this Project 1 (PI: J Byrd) plans to disrupt human B cell signaling in chronic lymphocytic leukemia with two novel compounds, examine innate immune effector cell function, and move to the clinic combining these agents with mAb B cell therapy; Project 2 (PI: S Tridandapani) will explore negative regulators of human monocytes / macrophage Fc? receptor signaling, as well as immune activators that reverse such negative regulators in order to improve antibody dependent cellular cytotoxicity (ADCC) of both liquid and solid tumors. Project 3 (PI: M Caligiuri) examines the mechanisms by which human natural killer (NK) cells acquire Fc?Rlll, and characterizes negative regulators of NK ADCC. Two clinical trials will then inhibit this negative signaling in order to enhance NK ADCC against lymphoma and NK cytotoxicity against multiple myeloma; Project 4 (PI: W Carson) examines the mechanism by which myeloid derived suppressor cells suppress innate effector cell ADCC, and explores different mechanisms to deplete these suppressor cells in order to enhance ADCC in the laboratory and in the solid tumor clinic. These four projects are served by the unique resources provided by Core A (Administration), Core B (Biostatistics) and Core C (Mouse Modeling and Animal Development). For the past 4.5 years, investigators within this P01 have published 51 collaborative manuscripts and have accrued 815 patients to clinical trials emanating from this P01. The next 5 years of work will result in improved clinical outcome of cancer patients undergoing with mAb therapy.

DESCRIPTION (Provided by applicant): The goal of the CALGB Leukemia Correlative Sciences Committee (LCSC) is to attain a highly successful integration of correlative laboratory studies into the design and implementation of CALGB leukemia clinical trials. During the current reporting period (6/1/02-5/31/08), members of this Committee discovered and/or validated multiple prognostic molecular and cytogenetic markers that can be used for risk-adapted patients'stratification into clinical trials and/or be regarded as therapeutic targets. The scientific progress made by the LCSC during the current reporting period is supported by 162 publications (70 manuscripts and 92 abstracts published or in press). To continue this work, in this competing renewal application, the CALGB LCSC is requesting support for three established Cores (Core A: Cytogenetics;Core B: Leukemia Tissue Banking;Core C: Administration) and four projects, each of which utilizes material from patients that are enrolled on CALGB leukemia treatment protocols. A common theme of this proposal is the integration of the established prognostic markers with newly discovered markers, including genome-wide gene copy alterations and gene and microRNA expression profiles, to dissect acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) into molecular subsets for which response to treatment and clinical outcome can be predicted. Furthermore, accumulating data indicate that certain subpopulations of AML cells have high proliferative potential and self-renewal capacity similar to normal stem cells and therefore could mediate resistance to anti-leukemia chemotherapy. This Committee will pursue for the first time a relatively novel strategy to test the relevance of abundance and gene profiles of these so-called leukemia stem cells (LSCs) to treatment response and clinical outcome of AML patients. Thus, with the support of the Cytogenetics core and the Leukemia Tissue Bank, each of the four projects will address scientific questions that correlate cytogenetic and molecular findings with leukemia patients'diagnosis, response to treatment and survival. The projects, led by outstanding leaders in the field of leukemia biology, diagnosis, and treatment are: Project 1: "Molecular characterization of adult AML" (PI Bloomfield/Marcucci,) Project 2: "Functional and genomic characterization of leukemia stem cells in AML" (PI Armstrong) Project 3: "Genome-wide analysis of adult ALL" (PI Downing) Project 4:"Molecular, biochemical, and immunologic studies of early state and symptomatic CLL" (PI Byrd).

In acute myeloid (AML) and lymphoid (ALL) leukemias, the MLL gene fuses with over 50 partner genes or can be rearranged as the result of a self-fusion creating the partial tandem duplication {MLL-PJD). We discovered the MLL-PTD and have investigated the MLL-PTD role in leukemogenesis and its prognostic impact. We were the first to report that MLL-PTD occurs more frequently in cytogenetically normal (CN)- AML and in AML with +11 and was associated with adverse prognosis. Although more recently we have shown that the outcome of MLL-PTD AML patients has improved with intensive treatment, most of them die of their disease, thereby underscoring the need for novel and more personalized treatment approaches. Interestingly, we observed MLL-PTD blasts from patients with dismal outcome often harbor additional adverse prognostic molecular markers, such as the FLT3 internal tandem duplication {FLT3 ITD) and that certain epigenetic aberrations are characteristically associated with the MLL-PTD. These data, therefore suggest that additional genetic and epigenetic hits are necessary for development of the MLL- PTD leukemia phenotype. In support of the multiple hits leukemogenic model, the Mil PTD as a single knocked-in defect in mice benignly alters hematopoiesis but does not induce leukemia, while it cooperates with the knocked-in Flt3 ITD defect to induce fatal AML. Reversion of epigenetic changes by hypomethylating agents and histone deacetylase inhibitors increases sensitivity of MLL PTD cells to chemotherapy. Based on these findings, therefore, we hypothesize that MLL PTD AML can be used as a model to elucidate the molecular mechanism(s) leading to multi-step leukemogenesis so that novel targeting therapies can be developed. To test our hypotheses, we collectively as a laboratory and clinical investigational team working together propose the following three aims: 1. To further characterize genetic and epigenetic mechanisms leading to MLL PTD AML using an Mil PTD/Flt3 ITD murine model; 2. To design novel therapeutic approaches targeting genetic and epigenetic aberrations in MLL PTD AML; 3. To perform an expanded Phase I clinical trial using a combination of DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors followed by intensive chemotherapy to assess safety and efficacy in adult patients with relapsed or refractory AML, particularly in patients with the MLL PTD. Our ultimate goal is to improve outcome of MLL PTD AML patients. We anticipate, however, that some of the discoveries derived from this project will also improve our understanding and our ability to treat other subtypes of AML. RELEVANCE (See instructions): We were the first to identify a type of acute myeloid leukemia (AML) characterized by a gene mutation called MLL PTD that is associated with short survival. We have created a mouse model of MLL PTD leukemia. We will use this model to understand the mechanisms through which MLL PTD causes leukemia and test novel therapies. Our laboratory discoveries will be rapidly moved to the clinic. Our goal is to improve the outcome of patients with MLL PTD AML, but also to extend our discoveries from to other subtypes of AML.

DESCRIPTION (provided by applicant): The Ohio State University Comprehensive Cancer Center (OSUCCC) is currently in its 35th year as an NCI designated CCC and is now requesting continued federal support for the next five years. Dr. Caligiuri currently continues in his seventh year as the OSUCCC Director and has since been named CEO of OSU's freestanding James Cancer Hospital. The overall goal remains to reduce cancer morbidity and mortality through continued basic, translational and clinical research. The 239 OSUCCC full, associate or introductory members are currently served by 18 shared resources and are distributed among the our six Research Programs which remain unchanged: Cancer Control, Experimental Therapeutics, Innate Immunity, Molecular Biology and Cancer Genetics, Molecular Carcinogenesis and Chemoprevention, and Viral Oncology. Since the last competitive renewal, the OSUCCC has shown significant growth as demonstrated by: 1) the recruitment of 159 faculty focused in basic, translational and clinical cancer research and medicine;2) more than a 80% increase in patient accrual to investigator-initiated trials;3) the addition of 5 new shared resources at an institutional investment of over $4.2 million;4) a 96% increase in total NCI funding despite a period of relatively flat federal funding;5) an 85% increase in publications, 51% of which were collaborative (i.e., inter-, intra-programmatic, or both);5) discovery, preclinical development and administration of two new anti-cancer agents into man, along with additional important advances in basic and clinical cancer research. The OSUCCC has also seen tremendous growth in institutional commitment since 2004 as demonstrated by 1) a ten-fold increase in annual financial support (now approximately $50 million) for the OSUCCC under the control of the Director;2) an additional $7.0 million of cash annually from OSU for research and infrastructure expansion;3) a formal direct reporting relationship to the Executive Vice President and Provost with complete oversight of the University-wide cancer funding initiatives, opportunities and cancer grant submissions;4) A seat on the University President's cabinet providing representation of the CCC at the highest level of the University;5) a six-fold increase in space currently under the sole control of the Director, including new additional dry and wet laboratory and office space for recruitment of additional faculty;6) a written commitment and approval by the OSU Board of Trustees for the expansion of the Cancer Program facilities that will more than double the current square footage under control of the OSUCCC Director in the next five years at a cost of approximately $800 million. With these new resource commitments in place, the OSUCCC is poised for continued significant growth and expansion in the next 5 years.

No abstract provided

In the past year (12/1/2008-11/30/2009), 326 patients were registered onto 26 high priority, innovative, feasibility (i.e., pre-phase I, pilot) and phase 1 institutional clinical interventions. These studies reflect the efforts of 19 different OSUCCC principal investigators (Table 2) and the scientific activities of four OSUCCC programs; Experimental Therapeutics (ETP), Cancer Control (CC), Molecular Carcinogenesis and Chemoprevention (MCC) and Immunology (IMM). Patients enrolled onto these protocols are managed by the Clinical Trials Office (CTO) utilizing protocol-specific-clinical research support. Based upon the current accrual statistics for investigator-initiated therapeutic clinical trials and an estimate of approximately 40 active clinical trial participants per clinical coordinator per year, we can justify a request for 5.0 FTEs for protocol-specific clinical support in year one of the CCSG renewal (see justification above). The CTO now manages 365 open clinical trials, including 55 therapeutic trials that were authored by OSUCCC investigators. Clinical investigators at the OSUCCC are encouraged to actively pursue funding for trials through external peer review mechanisms whenever possible. Thus, a number of these investigator-initiated trials (12) are supported though the OSU phase 1 grant (UOl CA76576) or the OSU NOI grant {m^ CM62207). Trials that are not supported by external funds are eligible for Protocol-Specific Research Support.

The essence of the OSUCCC resides in its 6 Research Programs and 18 Shared Resources. It is composed of 179 full members, 38 associate and 24 introductory associate members from 13 of the 18 University colleges. Its clinical facilities include the freestanding James Cancer Hospital (JCH) as well as Nationwide Children's Hospital and now OSU East Hospital where cancer care is provided to inner city residents. To coordinate and manage an organization of this magnitude, the University supports the position of a strong CCC Director, who reports directly to the chief academic officer of the University, the Executive Vice President and Provost. Organizational capabilities of the CCC have been considerably strengthened by President Gee's appointment of Dr. Caligiuri as CEO of The James Cancer Hospital; his reporting to the President of the University, as well as his new membership on the President's Cabinet, and his continued membership on the Medical Center Executive Committee. In this capacity, Dr. Caligiuri privately meets at least monthly with President Gee, twice monthly with Provost Alutto, and at least weekly with Sr. VP Gabbe. This ensures that the Cancer Program's oversight planning and evaluation are in place and integrated within the University's strategic initiatives. These meetings assure cancer representation in the major initiatives across the University and the health system, and keep the President, Provost and Sr. VP adequately advised of all activity within the OSUCCC, including The JCH. In a large and diverse university, this organizational structure allows the Director to take maximum advantage of OSU's strengths and capabilities in promoting the cancer program agenda.

PROJECT SUMMARY: The broad, long-term objective of this proposal is to understand how cancer evades and disarms the immune system at the molecular, cellular and organism level so this process can be interrupted for the successful treatment of cancer with monoclonal antibody (mAb) therapy. In the past 4.5 years, members of Project 3 have worked interdependently with other members of this P01 to produce 51 collaborative publications and accrue over 800 patients to therapeutic clinical trials that have directly emanated from our P01. Importantly, we have created impact in that we have advanced our understanding of mAb therapy for cancer and have introduced new reagents in man whose efficacy likely depends on the innate immune system. This P01 competitive renewal application continues to primarily investigate the human innate immune system and Project 3, while integrated with the efforts of Projects 1, 2 and 4, is focused on natural killer (NK) cells. During the past five years it has become clear that: 1) the affinity of the Fc receptor (R) for the Fc fragment of mAb can predict clinical outcome in the mAb therapy of certain cancers; 2) elimination of T regulatory (Treg) cells can enhance cytotoxic T cell adoptive therapy in patients with chemoresistant cancer; 3) blockade of CTLA-4, an inhibitory ligand on cytotoxic T cells, can improve survival in patients with melanoma; 4) avoiding the interaction of the killer immunoglobulin-like receptor (KIR) with its MHC Class I ligand, the NK cell can enhance long term survival in T cell depleted HLA-haploidentical bone marrow transplant for acute myeloid leukemia. We hypothesize that a better understanding of NK cell FcR expression and of NK cell tolerance will allow us to improve the outcome of cancer patients treated with mAb therapy. In the coming cycle, we will work to understand how developing NK cells acquire FcyRlll (CD16) on their cell surface and assess whether this process results in improved antibody dependent cellular cytoxicity (ADCC). Next we will characterize negative regulators of NK cell ADCC in an effort to block these and enhance ADCC of cancer. Finally, we will perform two Phase l/ll trials that will block the inhibitory KIR in an attempt to enhance tumor killing in man, one in combination with an anti-CD20 mAb for the treatment of lymphoma. The work outlined for Project 3 will be interdependent on Projects 1, 2 and 4 as well as Cores A, B, and C When completed, it will provide new insights as to how NK cells may be modulated in order to improve ADCC with mAb therapy in man. We expect our work to result in an improvement in disease free survival for cancer patients following the administration of mAb therapy.

PROJECT SUMMARY (See irstructlons); In this competing renewal P01 application, the Administrative Core will continue to provide centralized grant administration, communication processing, and budget management This Core will also serve to amalgamate the investigators, their experimental findings and their ideas, evaluation of research efforts and also to direct the summary efforts toward the Program outcome. The Specific Aims of the Administrative Core include the following: 1. Provide administrative services to the investigators. This includes the management of project supplies, filing, development of memos, meeting minutes and communications covering all operations, including publications. 2. Organize monthly meetings of the Program Steering Committee to facilitate effective communication and decision making to ensure scientific progress of the Program. 3. Organize an annual External/Internal Scientific Advisory Board meeting. The External and Internal Scientific Advisory Board shall review all experimental findings and clinical outcomes, help prioritize investigations, review the coordination of collaborations, and evaluate concepts that emerge from the studies. 4. Maintain program integration activities. This includes data sharing, rapid publication efforts, and identify and institute other novel activities critical to maintaining and strengthening the integration of the program. 5. Provide overall fiscal review, accounting, and real time budget analyses. This includes reports, verbal communications, reviews and forward-looking projections on expenditures.

DESCRIPTION (provided by applicant): The treatment for malignant glioma (glioblastoma) remains a challenge. Several experimental paradigms continue to be utilized in clinical trials, such as oncolytic viruses (OV), engineered or naturally occurring virus strains that replicate selectively in tumors. Oncolytic herpes simplex virus type 1 (oHSV) clinical trials have revealed its safety in humans with malignant gliomas, but more work is required to increase its efficacy. The investigators of this program project thus hypothesize that oHSV replication and dispersal in glioblastoma is curtailed by multiple oHSV-based and host-based barriers and responses during the very initial phases of viral infection and replication {aim 1). Understanding the nature of these barriers and responses would allow us to exploit both pharmacologic and genetic modalities to circumvent oHSV-based and host-based barriers and responses and increase the efficacy of malignant glioma virotherapy (aim 2). To achieve these aims, project 1 (PI: J. Glorioso) plans to re-engineer oHSV to redirect it towards glioma cell surface receptors while increasing its safety using microRNA-based translational controls of oHSV genes; Project 2 (PI: E.A. Chiocca) will characterize a novel mechanism of antiviral action within tumor cells that is based on one of the histone deacetylases, HDAC6, how it interacts with Interferon and how it shuttles post-entry oHSV towards lysosomes for xenophagy rather than nucleus for active replication; Project 3 (PI: B. Kaur) will determine how the stroma of the tumor microenvironment impedes oHSV dispersal and will utilize chondroitinase to counteract this; Project 4 (PI: M.A. Caligiuri) will show how the rapid NK cell activation against virally infected nervous system tumors is deleterious to therapy and will characterize the cellular and molecular effectors of this response. These 5 projects will be served by the unique resources provided by Core A (Biostatistics/ Administration: Chiocca/Fernadez) that provides biostatistical justification for al projects, Core B (oHSV Production: Goins) that provides all projects with the same stock of purified oHSV and Core C (Glioma Biorepository: Nakano) that provides all projects with patient-derived glioma spheroids (GSs) that recapitulate the human tumor phenotypic/genetic features.

The ultimate goal of this proposal is to understand the role of innate immunity within the context of oncolytic viral (OV) therapy for glioblastoma multiforme (GBM). OV treatment of GBM relies on cancer-specific replication of the virus leading to tumor destruction with minimal toxicity to adjacent non-neoplastic tissue. Results from the 5 clinical trials in patients with malignant glioma have shown the relative safety of this novel treatment modality. However, evidence for significant efficacy remains to be established. Our attention in this project focuses on the natural killer (NK) cell response following viral administration. Due to their antiviral properties, NK cells represent a potential barrier to OV therapy. Alternatively, the antitumor NK response has the potential of augmenting the tumor clearing properties of OV therapy. Our previously published rat in vivo studies have demonstrated that 1) the inflammatory response mediated by activated macrophages/microglia limit OV efficacy and 2) transient immune modulation with cyclophosphamide significantly enhances GBM clearance following OV inoculation. As a result, we hypothesize that NK cells coordinate a robust inflammatory response following initial OV administration that limits viral replication, spread, and tumor lysis, thereby creating a barrier to effective OV therapy for GBM. In this project, we will determine: a) how the antiviral and antitumor properties of NK cells impact overall OV therapeutic efficacy for GBM; b) how NK cells mediate macrophage/microglia inflammatory polarization; c) the key mechanistic signals that lead to NK mediated clearance of OV-infected glioma; and d) how pharmacological agents such as cyclophosphamide and valproic acid modulate the NK response following OV treatment, leading to enhanced clearance of GBM. By elucidating the role of NK cells in orchestrating the inflammatory response following OV therapy, we are highlighting a critical therapeutic target that, when hiodulated, will potentially enhance OV efficacy for GBM. Moreover, the significance of our experiments extends beyond virotherapy into the realm of virology and immunology as we attempt to discern the key mechanistic signals leading to NK mediated clearance of viral infection.

DESCRIPTION (provided by applicant): Dietary components have become very attractive agents for cancer prevention. Toward this end, we envision the use of dietary components to modulate innate immune cell function with the goal of cancer prevention. Natural killer (NK) cells can be modulated to cure acute myeloid leukemia (AML), and NK cell activity is correlated with relapse-free survival in AML. AML is a disease that primarily affects older adults: the median age at diagnosis is over 65, and the 5-year survival rate remains under 10%. Elderly AML patients are less able to tolerate therapies effective in younger patients, such as intensive chemotherapy and allogeneic stem cell transplantation. Therefore, novel, less toxic, approaches to prevent AML and AML relapse represent an unmet therapeutic need. Our preliminary data show that phyllanthusmin C (PL-C), a diphyllin lignan isolated from edible plants, can enhance NK cell activity. PL-C enhances human NK cell interferon-gamma (IFN-?) production via upregulation of the NF-kB signaling component, p65. This finding is consistent with our in vivo data, showing that treatment with PL-C can prolong the survival of the mice bearing AML in our syngeneic orthotopic animal model. We also found that PL-C can be detected in murine plasma 30 minutes after administration via oral gavage at a dose of 200 mg/kg. Based on these data, we hypothesize that PL-C originated from edible plants can enhance NK cell tumoricidal activity, and thus may possess therapeutic efficacy in the prevention of cancers including AML. We propose to test our hypothesis through the following three Aims: Aim 1 is to determine whether PL-C regulates NK cell cytotoxicity against AML cells in vitro; Aim 2 is to investigate the mechanisms by which PL-C enhances NK cell anti-tumor activity. Aim 3 is to study the preclinical efficacy of PL-C in the prevention of AML in vivo. To our knowledge, this is the first time that a lignan-based dietary component from an edible plant has been demonstrated to specifically and significantly enhance human NK cell function. As such, our study may open a new avenue for cancer prevention. Because PL-C is a compound that comes from an edible plant, this preclinical study has relatively strong potential for translation into the clinic as Phse I, Phase II, and then Phase III chemoprevention trials to be initiated following completion of this study. Therefore, not only is our study innovative and significant, but it is also highly translational.

? DESCRIPTION (provided by applicant): The NCTN's Alliance for Clinical Trials in Oncology (Alliance) represents a merger of three former NCI-funded cancer cooperative groups (ACOSOG, CALGB, and NCCTG). Combined, these groups and their associated biorepositories represent an unequaled resource of expertise, institutional infrastructure, and high quality, densely annotated patient biospecimens collected from several decades of cancer therapeutic trials. The scientific mission of the Alliance Biorepository and Biospecimen Resource (ABBR) is to support the activities of the Alliance, the NCTN, and the broader cancer research community through four specific aims, which are: 1) To prospectively support Alliance and NCTN-wide clinical cancer trials with respect to biospecimen procurement, tracking, processing, quality assurance, storage, and distribution. The ABBR will develop approaches to streamline biospecimen collection and support the collection of novel biospecimen types for genomic- and proteomic-based biomarker studies in the context of therapeutic trials; 2) To provide a resource of high quality, densely annotated biospecimens for secondary correlative science studies directed toward biomarker validation with high clinical impact. The ABBR will work with other NCTN biorepositories, the NCTN Biospecimen `Front Door Service', and the NCTN Navigator tool to provide inventories of biospecimens that may be suitable for secondary correlative science studies proposed both within the NCTN groups as well as the broader cancer research community. It will also work with other NCTN members and NCI to create an efficient and transparent process for reviewing, approving, and executing such requests so as to speed the translation of candidate biomarkers to cancer diagnostics with clinical utility; 3) To provide scientific leadership to the NCTN biobanking enterprise, by active participation in the NCTN Group Banking Committee (GBC). The ABBR will continue its outstanding commitment to harmonizing and improving biobanking Best Practices by active participation in GBC meetings, subcommittee's activities, and work products, and; 4) To provide expertise and infrastructure support for biobanking efforts beyond the NCTN, including NCI's Experimental Therapeutics Clinical Trials Network (ETCTN) and the Community Oncology Research Program (NCORP). By creating a federated infrastructure of biorepository sites and a unifying informatics platform for tracking biospecimens across them, the ABBR is uniquely poised to collaborate with other NCI programs, whenever biorepository or biospecimen resources are required for specific translational cancer research efforts.

CORE-002: DEVELOPMENTAL FUNDS PROJECT SUMMARY / ABSTRACT In the 2009 CCSG competing renewal, ?Developmental Funds? received a merit descriptor of ?outstanding to exceptional? noting that the use of the funds was extraordinary in terms of the quality of recruitments and leveraging of the developmental funds to the Center's mission. During the current funding period, the OSUCCC benefited from $1,625,283 of CCSG Developmental Funds. For the $1,086,942 of CCSG dollars used for faculty and staff recruitment, the return to-date in total direct costs for CCSG-approved peer-reviewed extramural funding is $45,841,496, a 42:1 return on investment, and in total funding was $47,389,764, a nearly 44:1 return on investment. In addition, we used CCSG funds in the development of two needed and well utilized Shared Resources (SRs), the Medicinal Chemistry SR and the Nutrient and Phytochemical SR, both of which have now transitioned from developing to full Shared Resources and are presented in this application. In the new funding period, we request $2,000,000 in CSSG Developmental Funds over 5 years and propose using this support to continue to focus on building and innovating within our three traditional areas strength: genetics, experimental therapeutics, and prevention. Specifically, we propose to identify and recruit 18 senior, midlevel and junior faculty whose scientific expertise targets relevant needs in the OSUCCC strategic plan and/or catchment area. Further, we propose the development of two shared resources, Solid Tumor Translational Science and the Veterinary Clinical Research Support, that are innovative and necessary to support the science of the Research Programs and their individual member investigators.

CORE-001: PLANNING AND EVALUATION PROJECT SUMMARY / ABSTRACT Rated as ?Exceptional? in 2010 by the NCI Review Team, it was noted that ?a variety of activities contributes to the Program Planning and Evaluation efforts of the OSUCCC and these activities have been very effective in increasing the success of the Cancer Center.? At the OSUCCC, the planning and evaluating approach provides a robust structure for continually assessing progress relative to our strategic plan, our peers, and the needs our of our catchment area and the National Cancer Institute. Further, planning and evaluation is a means for identifying opportunities for leveraging resources within the OSUCCC, the James Cancer Hospital, the Medical Center, the University and with other Institutions in support of our strategic goals to be a leader in transformative science that translates to a reduction in the burden of cancer locally, nationally and globally. Finally, having robust planning and evaluation processes allows us to function at a high level of awareness to effectively and efficiently modify our plans to adapt to the ever changing environment of science, technology, healthcare and funding.

ADMIN CORE-001: ADMINISTRATIVE CORE PROJECT SUMMARY / ABSTRACT Overall, Administration, rated as ?Outstanding? in the 2010 Summary Statement, holds the responsibility for managing and coordinating all financial and operational functions related to the strategic vision of the OSUCCC and CCSG. With Dr. Caligiuri assuming responsibilities as the OSUCCC Director as well as the CEO of the James Cancer Hospital in 2008, Administration was re-organized in 2010 in a similar fashion in order to integrate the administrative leadership to better facilitate the clinical and translational research mission of the OSUCCC. Further, the key roles that Administration now plays have grown considerably and are fully integrated across the Medical Center and the University to ensure the OSUCCC and CCSG needs are represented, prioritized and facilitated in an effective manner. Also in the 2010 Summary Statement, the reviewers rated Senior Leadership ?Exceptional?. Dr. Caligiuri now enters his 12th year as the OSUCCC Director and his 7th year as CEO of The James Cancer Hospital overseeing the entire cancer program at The Ohio State University. As the Director of the OSUCCC, Dr. Caligiuri has direct reporting to the Provost of the University and as CEO of the James Cancer Hospital he has direct reporting to the President of the University. To operate most effectively, Dr. Caligiuri has established a leadership structure with three key leadership positions directly reporting to him, the Senior Executive Director for Administration, the Deputy Director and the Physician-in-Chief. This executive team along with the Associate Directors of Basic Science, Clinical Research, Population Science, Biospecimen Research and Shared Resources as well as the Directors of Research Operations, Clinical Research Administration and Information Technology make up the core OSUCCC Senior Leadership Team. Now as the OSUCCC enters its 39TH year, the Senior Leadership team with Administration is responsible for coordinating the strategic growth of the Center, including facilitation of outstanding basic research programs, inter- and intra-programmatic collaboration, guiding translational and clinical research efforts, education and community outreach and provision of appropriate Shared Resource facilities. The team is also responsible for developing and executing long-term vision and strategic planning for the OSUCCC, and working towards the NCI goal to reduce morbidity and mortality from cancer. The changes that have occurred during the last funding cycle have been dramatic in terms of growth. Nonetheless, the path and vision set by the Director are for continued expansion of scientific discovery and translational research remains, with emphasis on our core strengths in cancer genetics, experimental therapeutics and prevention. Throughout this current cycle, Senior Leadership and Administration have worked together successfully to coordinate efforts and achieve several key accomplishments in support of the mission of the OSUCCC and CCSG.

The Molecular Carcinogenesis and Chemoprevention (MCC) Program, led by Steven K. Clinton, MD, PhD, has a collaborative team of 37 basic, translational and clinical scientists. These faculty have appointments in 17 Departments/Divisions within the Colleges of Medicine, Arts and Sciences, Pharmacy, Food Agriculture and Environmental Sciences, Public Health, Dentistry, Education and Human Ecology and Veterinary Medicine. The Specific Aims of the MCC Program are: 1) to characterize molecular and cellular changes induced by chemical, physical, hormonal and infectious agents that contribute to neoplastic transformation and multistage carcinogenesis in experimental models and humans; 2) to develop and characterize novel agents for cancer chemoprevention and define their efficacy, safety, and mechanisms of action using in vitro and preclinical models; and 3) to identify dietary and nutritional components that may enhance or inhibit the carcinogenesis cascade across the continuum of cancer progression. Each of these aims results in translational prevention studies in human populations with an emphasis on those at risk due to exposure to carcinogenic or cancer promoting agents, familial and genetic predisposition, or due to the presence of premalignant lesions. The MCC Program?s overarching goals, implemented through multiple MCC initiatives, are to accelerate the research objectives of each Aim through incentivizing and stimulating collaborative investigation among MCC members, other investigators of the OSUCCC as well as facilitating the implementation of translational studies of cancer etiology, prevention, and progression in human trials. The MCC enhances quality by promoting knowledge of and utilization of state-of-the-art technologies provided by the OSUCCC shared resources (members utilize 14/14 shared resources). The MCC Program, during its previous review (2004-2009) was graded as ?Outstanding to Exceptional?. During this funding period (2009-2014), MCC Program members published 484 cancer relevant peer-reviewed articles in top tier journals for the respective fields of carcinogenesis, chemoprevention, and nutrition. Collaboration is extensive with 28% intra-programmatic publications and 55% inter-programmatic publications, with 272 or 56% being multi-institutional and 447 or 92% being collaborative publications. Peer-reviewed funding for the MCC Program is $5.19M in annual direct costs with $2.9M (56%) from the NCI. Translational research has been robust as well with 20 human trials led by MCC members employing the OSUCCC Clinical Trials Office resulting in 360 interventional accruals during the last funding cycle, 72% of which were from investigator-initiated Phase I and II trials. The current MCC Program uniquely integrates investigators across disciplines yet with shared interests focusing upon the interactive themes of carcinogenesis, chemoprevention, and nutrition. Our future goals include the integration of new initiatives involving the microbiome and metabolomics, two areas benefiting from rapid growth in technology and bioinformatics that will dramatically impact our understanding of carcinogenesis and strategies for cancer prevention.

DESCRIPTION (provided by applicant): Dietary components have become very attractive agents for cancer prevention. Toward this end, we envision the use of dietary components to modulate innate immune cell function with the goal of cancer prevention. Natural killer (NK) cells can be modulated to cure acute myeloid leukemia (AML), and NK cell activity is correlated with relapse-free survival in AML. AML is a disease that primarily affects older adults: the median age at diagnosis is over 65, and the 5-year survival rate remains under 10%. Elderly AML patients are less able to tolerate therapies effective in younger patients, such as intensive chemotherapy and allogeneic stem cell transplantation. Therefore, novel, less toxic, approaches to prevent AML and AML relapse represent an unmet therapeutic need. Our preliminary data show that phyllanthusmin C (PL-C), a diphyllin lignan isolated from edible plants, can enhance NK cell activity. PL-C enhances human NK cell interferon-gamma (IFN-g) production via upregulation of the NF-kB signaling component, p65. This finding is consistent with our in vivo data, showing that treatment with PL-C can prolong the survival of the mice bearing AML in our syngeneic orthotopic animal model. We also found that PL-C can be detected in murine plasma 30 minutes after administration via oral gavage at a dose of 200 mg/kg. Based on these data, we hypothesize that PL-C originated from edible plants can enhance NK cell tumoricidal activity, and thus may possess therapeutic efficacy in the prevention of cancers including AML. We propose to test our hypothesis through the following three Aims: Aim 1 is to determine whether PL-C regulates NK cell cytotoxicity against AML cells in vitro; Aim 2 is to investigate the mechanisms by which PL-C enhances NK cell anti-tumor activity. Aim 3 is to study the preclinical efficacy of PL-C in the prevention of AML in vivo. To our knowledge, this is the first time that a lignan-based dietary component from an edible plant has been demonstrated to specifically and significantly enhance human NK cell function. As such, our study may open a new avenue for cancer prevention. Because PL-C is a compound that comes from an edible plant, this preclinical study has relatively strong potential for translation into the clinic as Phse I, Phase II, and then Phase III chemoprevention trials to be initiated following completion of this study. Therefore, not only is our study innovative and significant, but it is also highly translational.

ABSTRACT The traditional ?standard of care? for cancer over the past several decades has consisted of surgery, radiation therapy, chemotherapy, and stem cell transplantation in some cancers such as acute myeloid leukemia. In the last decade, immune therapy using monoclonal antibodies such as rituximab and traztuzumab have now been incorporated into the standard of care for lymphoma and breast cancer, respectively. Compelling data now shows that that innate immune cells, such as natural killer (NK) cells are an important component mediating the clinical effectiveness of antibody therapy. Our proposal aims to strengthen cellular innate immunity to enhance cancer therapy. Part of the challenge relates to 1) limited recognition of tumor cells by innate immune effector cells; 2) dampening of innate immune effector cell function in the tumor microenvironment; and 3) penetration of the tumor mass by innate immune effector cells. Our vision is to add activated, tumor antigen specific innate immune effector cell therapy to the existing armamentarium against both liquid and solid tumors. Strategically, this will be accomplished over the next 7 years through: 1) a more complete understanding human NK cell development, including the expression of activating and inhibitory receptors; 2) optimizing the development of human NK cells expressing chimeric antigen receptors, and 3) full development of our bi-specific NKG2D antibody that will bring human NK cells and other innate immune effector cells into the tumor bed. Our documented consistent success with basic research on NK cell development and effector function over decades, along with our successful translation of a multitude of our discoveries to the clinic in over 1,000 cancer patients both serve as a foundation for us to successfully proceed along this pathway. In the next seven years, we believe we will lead the field in: 1) enhancing NK cell tumor specificity; 2) enhancing NK cell cytolytic effector function; and 3) enhancing migration of NK and other innate immune cells into the tumor bed. The potential outcome over the next 7 years will be the completion of a series of clinical trials that first demonstrate significant anti-tumor activity in man and ultimately demonstrate significantly prolonged survival of cancer patients bearing liquid or solid tumors that express antigens to be recognized by our modified innate immune products.

A major area of institutional support for clinical cancer research is for investigator-initiated trials. The arrival of new investigators into the OSUCCC, the increased attention paid to translational research and changes in the funding landscape of clinical trials requires that there be a mechanism for funding investigator initiated studies of novel agents and/or novel design. The support of innovative Phase I trials and pilot studies was previously supported by the CCSG-mandated Protocol Specific Research Support mechanism. With the recent changes in the CCSG guidelines, the Early Phase Clinical Research Support (EPCRS) mechanism has been employed since late 2013 as a means of stimulating the conduct of early phase, investigator-initiated studies. The EPCRS process is overseen by Dr. William Carson, the Associate Director for Clinical Research (ADCR). Investigators are informed of this OSUCCC support mechanism via official communications from OSUCCC administration and the Disease Specific Research Group (DSRG) leaders. There is a formal application process which is conducted quarterly. Proposals are reviewed by a panel, chaired by the ADCR, Dr. William Carson. Successful applications are selected for CCSG support in the form of research nurse or data management assistance from the CCC supported Clinical Trials Office (CTO). On average 1-3 studies have been approved for support each quarter. In the current funding cycle, a total of 13 studies have been supported by the EPCRS and have accrued a total of 175 patients in the last year. In addition, to EPCRS, the OSUCCC has a formal, peer-reviewed mechanism, called the Intramural Research Program (IRP), to channel developmental funds from Pelotonia, our annual bicycle fundraising event, into bench to beside research.

The Protocol Review and Monitoring System [PRMS] of the OSUCCC consists of a Clinical Scientific Review Committee [CSRC] that reviews all new cancer-related clinical protocols for scientific merit prior to IRB submission and monitors the scientific progress of ongoing studies including accrual rates. The main CSRC is organized into two teams (with highly similar composition) that collectively meet twice monthly (each once per month), facilitating rapid protocol review. The CSRC consists of 37 members representative of all the OSUCCC research programs and includes clinical and basic researchers, biostatisticians, pharmacists, and research nurses. Additionally, members with specific expertise in the areas of cancer prevention and control as well as pediatrics are convened as CSRC sub-committees to provide focused review of these protocols. At its twice monthly meetings, the main CSRC performs full scientific reviews of all cancer-related clinical protocols initiated by local investigators or pharmaceutical industry sponsors and acknowledges NIH peer-reviewed studies. Each CSRC member is expected to be present at each scheduled meeting, and a majority of the voting members scheduled to be at a meeting must be present to attain a quorum for the meeting. Failure to achieve a quorum at a meeting precludes any business being conducted. In addition to achieving quorum, appropriate reviewer expertise must be present in order for business to be conducted. CSRC approval of a protocol is required prior to its review by the OSU Cancer Institutional Review Board. Each protocol is reviewed by four CSRC members (a primary reviewer, a secondary reviewer, a biostatistician, and a pharmacist). Reviewers follow a written review template that calls for an analysis of the scientific hypothesis and rationale, experimental design, patient inclusion and exclusion criteria, treatment plan, statistical plan, pharmacy considerations and proper prioritization. As of January 2009, all new research protocols undergo concurrent review by the CSRC Logistics Review Committee (LRC), which is a sub-committee of the CSRC. The LRC was implemented to facilitate trial activation and confirm proper research prioritization. The LRC does not evaluate the scientific aspects of the protocol, but does evaluate the availability of the resources necessary to successfully implement and complete the research protocol. The CSRC Executive Committee (EC) provides oversight to the CSRC. It is led by the CSRC Chair. The EC consists of eight CSRC members that meet monthly to review protocol accrual and scientific progress and ensure that protocol prioritization rules are followed. The CSRC EC is responsible for communication with investigators whose clincial trials are not meeting their accrual goals and providing support to facilitate accrual. The CSRC adheres to a set of welldefined criteria for accrual monitoring and trial prioritization. Those ongoing studies that do not show adequate accrual or fail to meet accepted standards of quality control based on formal audits are terminated. In 2014, the CSRC closed 18 trials for low accrual. The EC also performs expedited reviews for appropriate studies (e.g., retrospective studies). In the last year (12/1/13 to 11/30/14), 199 new protocols were reviewed by the CSRC with the following dispositions: 68 (34%) were approved as written, 55 (28%) were approved with stipulations, 14 (7%) were deferred, 57 (29%) were administratively acknowledged after having undergone previous NIH peer-review, and 5 (2%) were disapproved. Twenty-eight non-interventional protocols were also administratively approved by the CSRC EC.

The Translational Therapeutics Program at The Ohio State University Comprehensive Cancer Center (OSUCCC), led by David Carbone, MD, PhD and Denis Guttridge, PhD, unites an outstanding team of 55 basic, translational and/or clinical researchers from 17 departments within the Colleges of Business, Medicine, Engineering, Pharmacy, and Veterinary Medicine at Ohio State University. This Program is a continuation of the 15 year-old Experimental Therapeutics (ET) Program that included both liquid and solid tumors. However, as the result of a strategic effort to grow the solid tumor presence in the OSUCCC, we successfully recruited an additional 38 solid tumor physicians, basic scientists, and physician-scientists during this last funding cycle, 16 of who came into the ET Program. We also transitioned to new leadership in the departments of radiation oncology and pathology, as well as in the divisions of medical oncology, surgical oncology and gynecologic oncology, each with expertise in translational medicine and each a member of this Program. With more emphasis on translation and less emphasis on experimental pharmacology, we renamed the ET Program ?Translational Therapeutics (TT)?. Under the leadership of Drs. Carbone and Guttridge the TT Program has seen significant progress during the last cycle, producing 874 peer-reviewed publications among which 9% are in high impact (>10) journals, 28% from intra-programmatic collaborations, 43% from inter-programmatic collaborations; 59% are multi-institutional and 86% are collaborative publications. TT Program members have collaborated on programmatic grant submissions and have been awarded 2 NCI P01s, a U01, a U54 SPORE, and a U10 programmatic grant focused on solid tumor biology, as well as two T32 training grants. As a consequence of these and other collaborative efforts, the TT Program has $8.8M in current annual direct costs from peer-reviewed grants of which $7.3M (83%) is from the NCI. The TT Program is well-integrated with the clinical teams via participation in the multidisciplinary Disease Specific Research Groups. As such, there were 5,253 accruals to interventional clinical trials during the last funding cycle of which 4,357 (83%) were therapeutic; 2043 (47%) of the latter resulted from investigator-initiated clinical trials. The overall goal of the TT Program is to pursue solid tumor biology in order to develop and translate promising preclinical studies into innovative clinical trials for the successful prevention, diagnosis and treatment of solid tumors. This goal will be achieved by performing the following specific aims: 1) Identifying alterations in solid tumor signaling pathways to develop targeted therapeutics; 2) Identifying and therapeutically targeting tumor-host interactions; and 3) Improving upon or developing new approaches for determining prognosis, selecting appropriate therapy and evaluating the response to treatment. Future directions include continued strategic recruitments and heavy leveraging of the OSUCCC?s Drug Development Institute (DDI) to move a number of our exciting compounds in the TT Program into the clinic for a variety of solid tumors.

Rated as ?Exceptional? in 2010 by the NCI Review Team, it was noted that ?a variety of activities contributes to the Program Planning and Evaluation efforts of the OSUCCC and these activities have been very effective in increasing the success of the Cancer Center.? At the OSUCCC, the planning and evaluating approach provides a robust structure for continually assessing progress relative to our strategic plan, our peers, and the needs our of our catchment area and the National Cancer Institute. Further, planning and evaluation is a means for identifying opportunities for leveraging resources within the OSUCCC, the James Cancer Hospital, the Medical Center, the University and with other Institutions in support of our strategic goals to be a leader in transformative science that translates to a reduction in the burden of cancer locally, nationally and globally. Finally, having robust planning and evaluation processes allows us to function at a high level of awareness to effectively and efficiently modify our plans to adapt to the ever changing environment of science, technology, healthcare and funding.

The Molecular Biology & Cancer Genetics (MBCG) Program at The Ohio State University Comprehensive Cancer Center (OSUCCC), led by Michael Ostrowski, PhD and Matthew Ringel, MD, unites a highly productive, collaborative and cancer focused team of 44 basic and translational scientists representing 7 colleges and 18 academic departments at The Ohio State University. Program science is centered in four major cancer-focused scientific themes: small non-coding RNAs and cancer, human cancer genetics/genomics, signal transduction and therapeutic resistance, and tumor microenvironment. Program members utilize state-of-the-art approaches to 1) identify genes and pathways that fuel tumor cell initiation and growth, and 2) provide mechanistic details of how these genes and pathways contribute to tumor progression and therapeutic resistance. Our overall goal is to define the mechanisms that account for the association between genes and cancer and to exploit this knowledge in order to reduce the incidence of death from cancer. The Specific Aims of the MBCG Program are: 1) to identify human genes, including non-coding genes such as those encoding microRNAs, that either through direct mutations or epigenetic mechanisms, result in an increased predisposition to cancer; 2) to determine the molecular mechanisms underlying the expression and function of the genes contributing to normal development, cancer progression, and therapeutic resistance; 3) to utilize the knowledge gained from gene identification and gene functions in tumorigenesis in order to reduce the incidence of death from cancer. During the prior funding period of the OSUCCC P30 CCSG, MBCG Program members published 810 research papers, including 104 high impact (> 10) manuscripts in journals such as Cell, Cancer Cell, Science, Nature Medicine, Nature Cell Biology, Journal of Clinical Oncology, and Journal of Clinical Investigation. There is extensive collaboration, with 21% intra-programmatic, 31% inter-programmatic, and 66% multiinstitutional publications. Overall, 81% of MBCG publications are collaborative. MBCG Program members are principal investigators (PIs) on seven NCI programmatic grants, including a P01 and U01 in breast cancer (P01 CA097189, U01 CA154200), a P01 and P50 SPORE in thyroid cancer (P01 CA124570, P50 CA168505), a P01 in epigenetics (CA129242) and two U01s in lung cancer (U01 CA152758, U01 CA166905). They are PIs on CCSG-approved grants whose direct funding is currently $9.39 million, with $7.5 million in NCI funding (80% of total CCSG-approved funding). Program members are principal investigators on 21 active protocols that have accrued 5760 patients over the past five years. MBCG studies are predominantly interventional but non-therapeutic, and non-interventional as most of our interventional therapeutic work is accomplished collaboratively within the Translational Therapeutics and Leukemia Research Programs.

The Leukemia Research (LR) Program within The Ohio State University Comprehensive Cancer Center (OSUCCC) is co-led by two physician scientists; Drs. John C. Byrd and Michael R. Grever who bring together 34 peer-reviewed funded members from 9 Departments and 5 Colleges (Engineering, Pharmacy, Medicine, Veterinary Medicine, and Arts & Sciences). The program is focused on the etiology, pathogenesis, prognosis, and treatment of leukemia with emphasis on acute myeloid leukemia (AML), human T-cell lymphotrophic virus l (HTLV-1) leukemia, large granular lymphocyte leukemia, hairy cell leukemia and chronic lymphocytic leukemia (CLL). LR Program-wide seminars, retreats, and disease-specific meetings provide an avenue for interaction among scientists and physicians to improve the quality of scientific discovery made through research. The LR Program integrates the use of novel technologies and translational and clinical support personnel provided through OSUCCC shared resources to facilitate impactful discoveries that ultimately have the potential to change the lives of patients with leukemia. LR Program discoveries span diagnostics through therapeutics, many of which are actively tested in both investigator initiated clinical trials and also NCI sponsored Phase I and II as well as cooperative group trials where our members have prominent leadership roles. The Specific Aims of LR Program include: 1) to integrate genetic, epigenetic, RNA/protein, non-coding RNAs, biochemical and immunosuppressive features of leukemic cells in order to enhance risk stratification and target identification; and 2) to foster preclinical and clinical development of epigenetic, targeted molecular, and immunologic therapeutics directed at the causes of leukemic transformation and progression. LR Program members have published 557 manuscripts of which 48% are inter-programmatic, 53% are intraprogrammatic, 61% are multi-institutional; total collaborative publications are 93%. The Program has $8.7M in annual direct costs of peer-reviewed funding of which $6.1M (70%) is from the NCI, including strong programmatic funding in basic, translational, and clinical research. With regard to translational research, in five years the LR Program members have accrued 1,660 patients to interventional clinical trials, 100% of which were therapeutic accruals, and of those, 989, or 60%, were placed on investigator-initiated trials. Of the 1,660 patients accrued, 1,495 or 90% were on Phase I/II trials. LR Program members have led the development of two therapeutic agents (ibrutinib and idelalisib) that were recently FDA approved for leukemia. Additionally, LR Program members have developed two novel diagnostic tests that are now utilized in clinical practice. Our future plans consist of targeted recruitment, continued enhancement of collaborative efforts, better molecular classification of both AML and CLL for improvement in targeted therapies, and moving a number of preclinical and clinical observations onto novel clinical trials.

The Cancer Control (CC) Program at the Ohio State University Comprehensive Cancer Center (OSUCCC), led by Electra D. Paskett, PhD, MSPH and Mary Ellen Wewers, RN, PHD, MPH, has 52 members from 19 OSU departments within the OSU Colleges of Medicine, Public Health, Arts and Sciences, Law, Nursing, Education and Human Ecology, Dentistry, and Food, Agricultural and Environmental Sciences, as well as Nationwide Children?s Hospital and Cincinnati Children?s Hospital. Program members provide a wealth of behavioral, basic, clinical, policy, outcomes, and social scientific knowledge and expertise not only to the CC Program, but to the OSUCCC as a whole. The overall CC Program goal is to conduct research to reduce the incidence, mortality and morbidity of cancer, which is accomplished by employing a transdisciplinary research team approach. The Specific Aims of the CC Program are to: 1) Identify molecular, genetic, and behavioral factors related to cancer incidence and mortality; 2) Develop and test behavioral interventions to prevent or detect cancer early; and, 3) Assess and intervene on issues of cancer survivorship. An additional developing aim of the CC Program is to address policy in relation to cancer prevention, detection, and care. Within these aims, research focuses on cross-cutting themes including underserved/minority populations within our catchment area, communication research, tobacco use and toxicity (including regulatory science), and behavioral strategies that capitalize on our members? strengths, such as epidemiology, biology and behavior. The program now has 2 NCI P50-funded Centers, both showcasing not only our multi-level focus but also the transition from observational to interventional studies. In addition, many of our interventions, e.g., Patient Navigation and Bio-Behavioral Intervention to Reduce Stress after Breast Cancer, have been incorporated into clinical care. Moreover, our research has contributed to policy changes, most notably the establishment of a smoke-free campus at OSU. During this past funding period (2009 - 2014), the Program members published a total 611 articles in peer-reviewed journals of which 32% have intra-programmatic collaboration, 20% have inter-programmatic collaborations, and 404 (66%) of the publications represent multi-institutional collaborations. Many of these publications fall within each of these categories bringing the total collaborative publications to 85%. CC Program funding stands at $6.8M of which $6.2M is peer-reviewed funding and of that, $3.9M (63%) is NCI funding. CC Program investigators have enrolled 9,205 participants to research studies over the last 5 years, 64% to externally peer-reviewed studies and 34% to institutional studies, 1.2% to cooperative trial group studies and 0.8% to industry studies. Of these, 6,964 (76%) were enrolled on interventional studies and 2,241 (24%) on non-interventional studies. Future goals focus on increasing research in: 1) policy, allowing us to take advantage of health care policy initiatives; 2) molecular and genetic epidemiology and 3) patient outcomes, fostering more inter-programmatic research with clinicians.

The Clinical Protocol and Data Management (CPDM) function of the OSUCCC provides comprehensive resources for the conduct of cancer clinical research with an emphasis on the inclusion of women, minorities, children and other underserved populations. The CPDM is overseen by Dr. William Carson, the Associate Director for Clinical Research (ADCR). The cancer center?s Clinical Trials Office (CTO), supports the centralized administration of protocol development, implementation and conduct for all clinically active cancer research groups. No clinical cancer research may be conducted outside the CTO. Thus, the CTO oversees all cancer clinical trials and is the centralized organization for the conduct of such research. This team provides the trial administration, protocol tracking and monitoring, data management, regulatory processing and financial supervision necessary for the successful conduct of clinical trials in a methodologically-sound, expedient, and cost-effective manner. The CTO Medical Director is Steven Devine, MD and the CTO Medical Co-Director is Dr. Bhuvana Ramaswamy, MD. The CTO interim Administrative Director is Nancy Single, PhD. The OSUCCC Data and Safety Monitoring Plan outlines the structure and responsibilities of the Data and Safety Monitoring Committee (DSMC) which oversees and reviews the safe conduct of clinical research at this cancer center. The DSMC is headed by William Blum, MD. The inclusion of women, minorities and children in cancer clinical research is an important goal of this organization and effort in this area are coordinated by the Center for Cancer Health Equity under the leadership of Electra Paskett, PhD, OSUCCC Associate Director for Population Sciences.

In the 2009 CCSG competing renewal, ?Developmental Funds? received a merit descriptor of ?outstanding to exceptional? noting that the use of the funds was extraordinary in terms of the quality of recruitments and leveraging of the developmental funds to the Center?s mission. During the current funding period, the OSUCCC benefited from $1,625,283 of CCSG Developmental Funds. For the $1,086,942 of CCSG dollars used for faculty and staff recruitment, the return to-date in total direct costs for CCSG-approved peer-reviewed extramural funding is $45,841,496, a 42:1 return on investment, and in total funding was $47,389,764, a nearly 44:1 return on investment. In addition, we used CCSG funds in the development of two needed and well utilized Shared Resources (SRs), the Medicinal Chemistry SR and the Nutrient and Phytochemical SR, both of which have now transitioned from developing to full Shared Resources and are presented in this application. In the new funding period, we request $2,000,000 in CSSG Developmental Funds over 5 years and propose using this support to continue to focus on building and innovating within our three traditional areas strength: genetics, experimental therapeutics, and prevention. Specifically, we propose to identify and recruit 18 senior, midlevel and junior faculty whose scientific expertise targets relevant needs in the OSUCCC strategic plan and/or catchment area. Further, we propose the development of two shared resources, Solid Tumor Translational Science and the Veterinary Clinical Research Support, that are innovative and necessary to support the science of the Research Programs and their individual member investigators.

DESCRIPTION (provided by applicant): The treatment for malignant glioma (glioblastoma) remains a challenge. Several experimental paradigms continue to be utilized in clinical trials, such as oncolytic viruses (OV), engineered or naturally occurring virus strains that replicate selectively in tumors. Oncolytic herpes simplex virus type 1 (oHSV) clinical trials have revealed its safety in humans with malignant gliomas, but more work is required to increase its efficacy. The investigators of this program project thus hypothesize that oHSV replication and dispersal in glioblastoma is curtailed by multiple oHSV-based and host-based barriers and responses during the very initial phases of viral infection and replication {aim 1). Understanding the nature of these barriers and responses would allow us to exploit both pharmacologic and genetic modalities to circumvent oHSV-based and host-based barriers and responses and increase the efficacy of malignant glioma virotherapy (aim 2). To achieve these aims, project 1 (PI: J. Glorioso) plans to re-engineer oHSV to redirect it towards glioma cell surface receptors while increasing its safety using microRNA-based translational controls of oHSV genes; Project 2 (PI: E.A. Chiocca) will characterize a novel mechanism of antiviral action within tumor cells that is based on one of the histone deacetylases, HDAC6, how it interacts with Interferon and how it shuttles post-entry oHSV towards lysosomes for xenophagy rather than nucleus for active replication; Project 3 (PI: B. Kaur) will determine how the stroma of the tumor microenvironment impedes oHSV dispersal and will utilize chondroitinase to counteract this; Project 4 (PI: M.A. Caligiuri) will show how the rapid NK cell activation against virally infected nervous system tumors is deleterious to therapy and will characterize the cellular and molecular effectors of this response. These 5 projects will be served by the unique resources provided by Core A (Biostatistics/ Administration: Chiocca/Fernadez) that provides biostatistical justification for al projects, Core B (oHSV Production: Goins) that provides all projects with the same stock of purified oHSV and Core C (Glioma Biorepository: Nakano) that provides all projects with patient-derived glioma spheroids (GSs) that recapitulate the human tumor phenotypic/genetic features.

PROJECT SUMMARY Chimeric antigen receptor (CAR)-engineered T cell therapy has revolutionized treatment for certain B cell malignancies, but similar successes for acute myeloid leukemia (AML) have not yet been shown, although interim results of early phase clinical trials, including our own, are promising. We have constructed a CAR that targets the type III receptor tyrosine kinase (RTK) called FLT3. Utilizing a patient-derived xenograft (PDX) model of FLT3(+) human AML, we have shown that infusion of human FLT3-CAR T cells prolonged survival of AML- bearing mice in the absence of other therapies. Further, FLT3-CAR T cells did not affect engraftment or survival of hematopoietic stem cells (HSCs) in mice bearing AML. The effectiveness of FLT3-CAR T cell therapy in AML will depend on (1) surface antigen density of FLT3 on AML blasts, including leukemic stem cells (LSC) relative to normal HSCs, and (2) the ability of other immune effector cells to contribute to the eradication of AML in vivo. To this end, we have discovered that treatment of AML blasts with an RTK inhibitor (TKI) upregulates the expression of FLT3 on the AML blast and the LSC in vivo, relative to FLT3 expression on normal HSCs. To advance a second cellular therapeutic intervention for AML, we have expressed the FLT3-CAR in human natural killer (NK) cells to generate FLT3-CAR NK cells, which demonstrated potent anti-leukemic activity against FLT3(+) AML. Collectively, these discoveries have led us to conclude that a program directed against FLT3(+) AML has strong rationale, is innovative and could result in a significant decline in mortality for a subset of AML patients. Therefore, the long-term objective of this proposal is to perform both preclinical and clinical studies that will best define an optimal strategy to reduce mortality from AML with FLT3-CAR cellular therapy, either alone or in combination with TKIs. Our central hypothesis is that targeting relapsed/refractory FLT3(+) AML with FLT3-CAR T cells or FLT3-CAR NK cells in combination with TKIs will improve outcomes in AML. In this proposal, we will assess the feasibility, safety and toxicity of performing a phase I study of human FLT3-CAR T cell therapy directed against FLT3(+) AML (Aim 1), we will determine the mechanism by which TKIs upregulate the surface density expression of FLT3 on LSCs and HSCs (Aim 2), and we will optimize a FLT3-CAR NK cell platform and assess its functionality against AML alone or combined with TKI (Aim 3). To accomplish these objectives, we have begun clinical manufacturing of FLT3-CAR T cells to treat eligible patients with refractory or relapsed AML; we will utilize both human AML cell lines and patients' AML blasts in vitro as well as in vivo along with our PDX model for our correlative and preclinical studies evaluating FLT3-CAR T and FLT3-CAR NK cells in combination with TKI. Upon conclusion, we will understand how best to optimize cellular immune therapy to cure AML. Further insight into this process, as will result from the implementation and completion of this proposal is impactful as it will ultimately lead to a reduction in mortality for select patients suffering from AML.