Richard Ford, Ph.D. - US grants

Hodgkin's disease (HD) is a common lympheid neoplasm, chiefly of adolescents and young adults, but also seen in later life. Although great therapeutic strides have been made clinically in this form of cancer, little information on the biological and immunopathelogic characteristics of HD has been reported. Two major questions in HD remain: (1) the nature of the malignant cell and its relationship to the other "reactive" cells present in the polymorphous Hodgkin lesion, and (2) the cause of the characteristic cellular (T cell) immunologic defect seen in HD. We have recently hypothesized, based on our preliminary studies, that HD is a neoplasm of functional cells in the monocyte/macrophage lineage. We have shown that the putative neoplastic cells (Reed-Sternberg cells (RSC) and their mononuclear variants (HDC)) have markers consistent with this cellular origin and make soluble biologically active products (monokines) that can account for the histopathologic appearance of HD. These studies will focus on the further delineation of the phenotypic characteristics of RSC and HDC, the behavior of these cells in vitro, and their sensitivity to growth-promoting lymphekines secreted by immunoregulatory cells in their environment. We will attempt to establish long-term cell lines from the malignant cells. The second phase of the research will be to study the HD T-cell defect vis-a-vis the interleukin system of lymphoidgrowth factors. Since these growth factors have been shown to control and regulate human T-cell function, we will evaluate the HD patients' ability to generate and react to interleukin growth factors to ascertain if the characteristic T-cell immunodeficiency can be correlated with an intrinsic defect in the interleukin system. These studies should provide important new data on two central issues in HD. (IP)

High grade B cell non-Hodgkin's lymphoma (NHL-B) is a major neoplastic sequelae of infection with the AIDS virus, HIV-1. AIDS-related lymphomas (ARL) have been considered to be secondary to the immune deficiency state induced by HIV, although the direct involvement of HIV or other viruses such as EBV have not as yet been demonstrated. We have recently studied two patients with ARL that presented with a previously unreported histopathologic "syncytial variant" of Burkitt's (small non cleaved B cell) lymphoma. Cell lines were derived from these ARL which have been shown to contain retroviral particles on ultrastructural analysis, and reverse transcriptase (RT) activity in culture supernatants. In this proposal, we will isolate, characterize, and molecularly clone the ARL-associated human retroviruses (ARL-RV). It will be determined if the ARL-RV are related to the HIV subfamily of non-transforming lentiviruses associated with immune deficiency or possibly to the transforming HTLV subfamily oncoviruses. Alternatively, these viruses could represent previously unknown human retroviruses or represent recombinant retroviruses involving known and/or unknown retroviruses. We will also determine the species and host range of infectivity for normal and neoplastic cells in vitro. The SCIDHu murine model will be utilized in vivo to assess the potential of the ARL-RV for inducing neoplasia and/or immune deficiency. Other studies will determine the biologic activity (CPE, syncytia formation, cell immortalization in vitro, etc.) associated with these viruses. Finally, we will utilize the molecular clones and immunologic regents generated in our studies to determine the presence and distribution of the ARL-RV in ARL, non-AIDS NHL- B, and other populations believed to be at risk for lymphoma including iatrogenically immunosuppressed organ transplant recipients, intravenous drug abusers, etc. These studies should indicate whether the ARL-RV can play a significant role in the pathogenesis of ARL, or possibly in lymphomagenesis in general, and provide at least a preliminary indication of the presence of this retrovirus in various subsets of the human population.

The acquired immune deficiency syndrome (AIDS) is caused by the HIV-1 retrovirus, which significantly impairs the function of the human immune system causing immunocompromise and susceptibility to opportunistic infections (OI). The loss of immune function in AIDS gives rise to an increased propensity to develop lymphoid neoplasia, particularly high grade B cell non-Hodgkin's lymphomas (NHL-B). Development of NHL-B was observed early in the AIDS epidemic, but the number of AIDS-related lymphomas (ARL) has risen sharply in the last few years, possibly due in part to better therapy with drugs such as AZT, with the incidence approaching 10% of AIDS cases in some areas of the US. ARL have proven extremely difficult to treat, due not only to the aggressive biological nature of the lymphoma cells, but also due to the immunocompromised state of the patients often precluding treatment with the aggressive chemotherapeutic regimens used to treat this type of lymphomas. This proposal will launch a multidisciplinary study group, the Houston AIDS Lymphoma Study Group (HALSG), consisting of experienced clinicians involved in the treatment of AIDS and AIDS-relater lymphoma, pathologists involved in ARL diagnosis, and basic scientists with interests and expertise in pertinent areas of immunology, molecular biology, and genetics. This interdisciplinary group will meld new approaches to the clinical treatment of ARL with laboratory studies, designed to better define and characterize the biological and genetic nature of the ARL. A new treatment protocol for ARL is presented that addresses the need for less aggressive therapy in severely immunocompromised patients, in concert with immunorestorative treatment with the biological response modifier, Thymosin, to increase T cell responses in AIDS patients. Patients with a lesser immune deficit will be treated with more standard chemotherapy, which has provided promising preliminary results in such patients. All patients will be maintained on AZT to retard the progression of HIV disease. These clinical studies will be integrated with laboratory studies to delineate the possible role of a variety of immune parameters and the aberrant expression of certain proto- oncogenes such as c-nyc and p53, that may have clinical significance in ARL cells. Cytogenetic studies, using both classical and the newer techniques will be used to identify non-random chromosomal abnormalities observed in ARL, and their possible involvement with oncogene expression.

This project involves the training of graduate students in cultural anthropology at the University of Michigan. This is one of two graduate programs funded in a new competition for support for ethnographic research training. The program will use the funds to support the methodological training of cultural anthropology students before their critical dissertation fieldwork. These training activities will be suited to the individual student's needs, in research training with a faculty mentor or as pilot research to prepare for future dissertation fieldwork. This research training is important in order to maintain and develop the "state-of-the-art" of cultural anthropology's ethnographic research methods. The projects will upgrade the methodological training of cultural anthropologists, which will maintain the intellectual quality of this research area.

9354776 Fisher This proposal articulates the development of a new undergraduate mathematics education program consisting of three new courses for undergraduate preservice secondary mathematics teachers and two new courses in developmental entry level mathematics based on proven secondary reform curriculum. The developmental courses are adaptations of the highly acclaimed reform secondary mathematics curriculum, the Interactive Mathematics Project (IMP). The preservice courses provide training in the philosophies, attitudes, objectives, and methods underlying the new reform curriculum and include specific training in the delivery of the adapted IMP materials. The preservice teachers participate in a highly structured field experience based on reform ideas as they actually teach (under the supervision of mathematics education faculty), the two new developmental courses to college students with entry level mathematics deficiencies. The project is conducted in two distinct settings: CSU, Chico (a typical four year institution that has remedial students and a teacher preparation program) and in a collaborative setting between the University of California, Irvine (a research institution with preservice teachers) and Irvine Valley College (a community college that has a large population of remedial students). Development of the curriculum, materials, and implementation design allows easy export to either higher educational setting throughout the nation. This innovative preservice-developmental proposal provides both an important advancement in aligning secondary mathematics teacher preparation programs with reform ideas and an economical solution to the growing bottleneck of students with entry level mathematics problems. ***y

AIDS-related lymphomas (ARL) are a group of immune deficiency-associated, aggressive B cell, non-Hodgkin's lymphomas (NHL-B). The incidence of ARL in the US is currently increasing, as AIDS patients live longer, and become at greater risk for the later consequences of immune compromise. The biology of ARL, that primarily results in the small non-cleaved cell (SNCC; Burkitt's (BL)) or Large cell (Immunoblastic) histo-types, seems to involve a number of the pathogenetic programs involved in aggressive NHL-B in the HIV negative patient population, However, the rapidity of development and the prodigious cell growth potential of ARL, suggests that additional genetic, biologic, or immune factors may be involved. The biologic or molecular basis of the aggressive malignant B cell growth in the ARL is unknown. ARL arises in the B lymphoid cell lineage, after the accumulation of a variable number of specific oncogene, viral, or tumor suppressor gene abnormalities that lead to cellular immortalization in NHL-B precursor cells. Studies will pursue our preliminary results that suggest that the immortalized, NHL precursor B cells, aberrantly express one or more of the BCGF (IL14, LMW-BCGF) cytokine genes, probably as part of the malignant transformation process. inappropriately expressed BCGF genes in ARL become autocrine growth factors (AGF) for the lymphoma cells, that also constitutively express the IL-I4R. Co-stimulatory factors for normal B cell proliferation, such the CD4O ligand (CD4OL) gp39, will also be evaluated for growth stimulatory potential in the ARL. Other cytokines with putative B cell stimulatory activity, such as IL6 or lL1O, will also be studied for a possible role in mediating or co-stimulating ARL cell growth in vitro. Since autocrine growth in the ARL appears to be mediated by BCGF molecules, we will explore methods for inhibiting cell growth thru growth factor deprivation, by inhibiting these cytokine growth factor genes in the ARL, with cytokine inhibitory drugs, such as cyclosporin A (CsA), and anti-sense oligonucleotides (ASO). Pathogenesis of the ARL will be explored in vitro, in experimental B cell models, containing transfected oncogenes and growth factor genes, that will be extended to in vivo studies, utilizing immune deficient. SCID mice. Finally, we will seek to identify additional (or different) genetic lesions in the ARL, possibly accounting for the accelerated development and/or aggressive clinical behavior, using comparative genomic hybridization (CGH), for identifying chromosomal abnormalities, not identifiable using conventional cytogenetic techniques.

This proposal outlines an interdisciplinary traineeship program for the study of cognition and environmental contexts. The program brings together psychologist, anthropologists, and other social scientists who are interested in the scientific study of graduate students who will be knowledgeable about both methods and theory in both anthropology and cognitive science. The innovative structural components include psychology, participation in interdisciplinary research seminars meeting weekly throughout their graduate careers and participation in research at several field sites and collaborative research installations already established in locations in many parts of the world by University of Michigan scientists. the students trained by such a program are expected by be uniquely qualified to develop a scientific understanding of cultural aspects of cognitive structures and processes.

The Tissue Procurement and Banking Facility (TPBF) is an established CCSG- supported core facility at the MDACC, that provides peer-funded basic science and clinical investigators at this institution with human tumor tissues that have been removed for biopsy or therapeutic resections, for on-going research projects. The TPBF functions to procure virtually all pathologic tumor tissue from patient procedures, that is not used for initial diagnosis, follow-up diagnostic studies, including cryogenically stored for future clinical follow-up. This tumor tissue is obtained to provide fresh, sterile (if requested), well-characterized human tumor specimens expeditiously (, 1 hour), to the wide spectrum of research scientists involved in virtually every type of human tumor investigation. Special preservation procedures will be instituted for nucleic acids and other methodologies for genomic analysis. The TPBF, that stores and triages tumor tissue for disease-site tumor programs, as well as cryopreservation, is being expanded into new, specifically designated and appropriate out-fitted space, to provide for more efficient and inclusive tumor banking facilities. The expanded banking facility will not only provide state-of-the-art cryopreservation facilities, but will also include an interactive database that will provide all pertinent information on tumor specimens procured by the TPBF. It will also interface with other institutional databases (hospital, pathology, epidemiology, etc.) that can provide vital and important data and other pertinent information to the searcher such as confidentiality/security. The specific aims of this proposal involve: 1) providing a flexible specialized tissue procurement resource providing for the efficient and expeditious delivery of freshly-obtained, well characterized tumor tissue to requesting investigators; 2) to develop a contemporary, centralized institution-wide tumor tissue procurement and banking core facility which will provide tissue compatible with current and predicted methodologies for analysis of DNA, RNA and protein as well as other methods of analysis; 3) to develop an interactive, accessible computer database which will provide epidemiology, family history, patient treatment and patient outcome and 4) to coordinate with existing individual "satellite", programmatically administered and funded tissue banks currently operational within the institution.

Under the direction of Dr. Richard Ford, Mr. Severin Fowles will collect data for his doctoral dissertation. Through a combination of archaeological excavation and the analysis of extant museum materials he will reconstruct the social organization and population movements of prehistoric peoples in the Taos District of New Mexico during the several centuries preceding Spanish arrival. During this period of marked social change aggregated populations dispersed and then rearranged themselves in new concentrations marked by large multi-story pueblo villages. Some areas such as Taos which had been sparsely inhabited saw the rise of multiple village communities. Mr. Fowles wishes to understand both the origins and social organization of these entities.

Ethnographic data indicates that many early historic Southwestern Native American peoples were organized into segmentary lineage systems. Clearly defined subgroups existed within the society and individual membership was parentally inherited. Relationships between lineages were clearly defined and social control, marriage patterns, inheritance and ritual responsibility all were determined by lineage structure. While likely this pattern existed prehistorically, this is difficult to prove. It is also unclear how it originated. Mr. Fowles hypothesizes that the aggregated community he plans to examine was structured according to segmentary lineage principles, that its founders derived from distinct cultural groups who migrated from different areas and came together to found it, and that the individual lineages correspond to individual founding groups. To examine this model, will use excavated material from several museums to reconstruct population structure prior to aggregation and search for material signatures which characterize individual groups. He will then examine one large site, Pot Creek Pueblo to determine whether these antecedent populations came together to form the site. He will study the distribution of these cultural markers within the site to find out whether they blend or maintain clear and distinct associations and, if so, whether the are spatially segregated within the as one would predict if segmentary lineages were present.

This research is important for several reasons. It will provide data of interest to many archaeologists. It will provide insight into how groups at a technologically simple level adapt and rearrange themselves across a changing landscape and how they are organized into coherent and functioning societies. This project will also assist in training a promising young scientist.

DESCRIPTION (provided by applicant): Non-Hodgkin's Lymphomas (NHL) are common tumors of the human immune system, primarily of B cell lineage (NHL-B) that have been showing significant unexplained increases in incidence for the last three decades. Unlike normal B-lymphocytes aggressive forms of NHL-B show rapid, dysregulated B lymphocyte growth characteristics, while retaining typical B cell immuno-phenotypes, including expression of characteristic CD40 and SIg. cell surface receptors. Normal B cells, involved in inflammatory, or other immune functions, transduce signals to activate and release, the key transcription factor, NF-kappaB from its cytoplasmic inhibitor; but aggressive NHL-B cells, such as Large B cell Lymphomas (LBCL), show constitutive expression of nuclear NF-kappaB. Our studies have shown that this is accomplished by continually maintaining an assembled, scaffold-like, signaling platform as a concatenate molecular aggregate, called a Signalosome within a lipid raft microdomains, contained within or subjacent to the lymphoma cell membrane. We have developed a hypothetical model of aggressive NHL-B cell pathophysiology that envisions dysregulation of the CD40 mediated signaling pathways as the major mechanism controlling tumor cell growth and other parameters of malignancy. The CD40 Signalosome appears to be initiated through autochthonous production and cognate ligand binding of CD154 (CD40L, gp39) to the CD40 receptor on the lymphoma cell surface. Our studies have indicated that the necessary and sufficient conditions for CD154 expression seem to differ in NHL-B cells from normal activated T lymphocytes, suggesting that the neoplastic lymphocytes show dysregulated gene expression and signaling pathways to mediate autonomous tumor cell growth. Constitutive expression of NF-kappaB in NHL-B can be down-regulated by treatment with specific antibodies or antisense oligos to CD40 or CD154, that disrupt the integrity of the CD40 Signalosomes, resulting in the inhibition of lymphoma cell growth, and the induction of lymphoma cell death in vitro. In this proposal, we will study the molecular characteristics of the CD40 Signalosome, by isolating and sequencing the component proteins of the canonical CD40/NFkB pathway contained within the signalosome, and demonstrate the signaling capabilities of this macromolecular structure. We will also study the signaling pathways controlling gene expression of the CD40 ligand, CD154, that we believe is intimately involved with driving the signalosome pathway, to ascertain the mechanism of its "ectopic" expression, as well as its possible role the aggressive growth pattern and resistance to cell death shown by aggressive NHL-B. In addition to these abnormalities involving lymphoma cell proliferation and cell viability, NHL-B cells also show aberrant expression of the TGF-beta/SMAD-SNO/SKI system, that is responsible for negative growth regulation in normal B lymphocytes (as well as many other functions in most cell types). Our preliminary studies have shown that abnormalities in SNO/SKI gene expression are involved in abrogating the negative regulatory influence of this signaling system in NHL-B that also appears to be linked to the CD40/NFkB system in NHL-B. Our studies will further examine the role of this system in potentiating the biologic, and possibly the clinical aggressiveness of these lymphoid tumors, and explore whether inhibition of the dysregulatory elements in this pathway can have therapeutic potential. We believe that our CD40 Signalosome model will provide an important "roadmap" into the important control mechanisms involved in NHL-B, that can be used for developing new therapeutic approaches for this very important group of human lymphoid neoplasms.

DESCRIPTION (provided by applicant): Non-Hodgkin's Lymphomas (NHL) are common tumors of the human immune system, primarily of B cell lineage (NHL-B) that have been showing significant unexplained increases in incidence for the last three decades. Unlike normal B-lymphocytes aggressive forms of NHL-B show rapid, dysregulated B lymphocyte growth characteristics, while retaining typical B cell immuno-phenotypes, including expression of characteristic CD40 and SIg. cell surface receptors. Normal B cells, involved in inflammatory, or other immune functions, transduce signals to activate and release, the key transcription factor, NF-kappaB from its cytoplasmic inhibitor; but aggressive NHL-B cells, such as Large B cell Lymphomas (LBCL), show constitutive expression of nuclear NF-kappaB. Our studies have shown that this is accomplished by continually maintaining an assembled, scaffold-like, signaling platform as a concatenate molecular aggregate, called a Signalosome within a lipid raft microdomains, contained within or subjacent to the lymphoma cell membrane. We have developed a hypothetical model of aggressive NHL-B cell pathophysiology that envisions dysregulation of the CD40 mediated signaling pathways as the major mechanism controlling tumor cell growth and other parameters of malignancy. The CD40 Signalosome appears to be initiated through autochthonous production and cognate ligand binding of CD154 (CD40L, gp39) to the CD40 receptor on the lymphoma cell surface. Our studies have indicated that the necessary and sufficient conditions for CD154 expression seem to differ in NHL-B cells from normal activated T lymphocytes, suggesting that the neoplastic lymphocytes show dysregulated gene expression and signaling pathways to mediate autonomous tumor cell growth. Constitutive expression of NF-kappaB in NHL-B can be down-regulated by treatment with specific antibodies or antisense oligos to CD40 or CD154, that disrupt the integrity of the CD40 Signalosomes, resulting in the inhibition of lymphoma cell growth, and the induction of lymphoma cell death in vitro. In this proposal, we will study the molecular characteristics of the CD40 Signalosome, by isolating and sequencing the component proteins of the canonical CD40/NFkB pathway contained within the signalosome, and demonstrate the signaling capabilities of this macromolecular structure. We will also study the signaling pathways controlling gene expression of the CD40 ligand, CD154, that we believe is intimately involved with driving the signalosome pathway, to ascertain the mechanism of its "ectopic" expression, as well as its possible role the aggressive growth pattern and resistance to cell death shown by aggressive NHL-B. In addition to these abnormalities involving lymphoma cell proliferation and cell viability, NHL-B cells also show aberrant expression of the TGF-beta/SMAD-SNO/SKI system, that is responsible for negative growth regulation in normal B lymphocytes (as well as many other functions in most cell types). Our preliminary studies have shown that abnormalities in SNO/SKI gene expression are involved in abrogating the negative regulatory influence of this signaling system in NHL-B that also appears to be linked to the CD40/NFkB system in NHL-B. Our studies will further examine the role of this system in potentiating the biologic, and possibly the clinical aggressiveness of these lymphoid tumors, and explore whether inhibition of the dysregulatory elements in this pathway can have therapeutic potential. We believe that our CD40 Signalosome model will provide an important "roadmap" into the important control mechanisms involved in NHL-B, that can be used for developing new therapeutic approaches for this very important group of human lymphoid neoplasms.

DESCRIPTION (provided by applicant): Non-Hodgkin's Lymphomas (NHL) are common tumors of the human immune system, primarily of B cell lineage (NHL-B) that have been showing significant unexplained increases in incidence for the last three decades. Unlike normal B-lymphocytes aggressive forms of NHL-B show rapid, dysregulated B lymphocyte growth characteristics, while retaining typical B cell immuno-phenotypes, including expression of characteristic CD40 and SIg. cell surface receptors. Normal B cells, involved in inflammatory, or other immune functions, transduce signals to activate and release, the key transcription factor, NF-kappaB from its cytoplasmic inhibitor; but aggressive NHL-B cells, such as Large B cell Lymphomas (LBCL), show constitutive expression of nuclear NF-kappaB. Our studies have shown that this is accomplished by continually maintaining an assembled, scaffold-like, signaling platform as a concatenate molecular aggregate, called a Signalosome within a lipid raft microdomains, contained within or subjacent to the lymphoma cell membrane. We have developed a hypothetical model of aggressive NHL-B cell pathophysiology that envisions dysregulation of the CD40 mediated signaling pathways as the major mechanism controlling tumor cell growth and other parameters of malignancy. The CD40 Signalosome appears to be initiated through autochthonous production and cognate ligand binding of CD154 (CD40L, gp39) to the CD40 receptor on the lymphoma cell surface. Our studies have indicated that the necessary and sufficient conditions for CD154 expression seem to differ in NHL-B cells from normal activated T lymphocytes, suggesting that the neoplastic lymphocytes show dysregulated gene expression and signaling pathways to mediate autonomous tumor cell growth. Constitutive expression of NF-kappaB in NHL-B can be down-regulated by treatment with specific antibodies or antisense oligos to CD40 or CD154, that disrupt the integrity of the CD40 Signalosomes, resulting in the inhibition of lymphoma cell growth, and the induction of lymphoma cell death in vitro. In this proposal, we will study the molecular characteristics of the CD40 Signalosome, by isolating and sequencing the component proteins of the canonical CD40/NFkB pathway contained within the signalosome, and demonstrate the signaling capabilities of this macromolecular structure. We will also study the signaling pathways controlling gene expression of the CD40 ligand, CD154, that we believe is intimately involved with driving the signalosome pathway, to ascertain the mechanism of its "ectopic" expression, as well as its possible role the aggressive growth pattern and resistance to cell death shown by aggressive NHL-B. In addition to these abnormalities involving lymphoma cell proliferation and cell viability, NHL-B cells also show aberrant expression of the TGF-beta/SMAD-SNO/SKI system, that is responsible for negative growth regulation in normal B lymphocytes (as well as many other functions in most cell types). Our preliminary studies have shown that abnormalities in SNO/SKI gene expression are involved in abrogating the negative regulatory influence of this signaling system in NHL-B that also appears to be linked to the CD40/NFkB system in NHL-B. Our studies will further examine the role of this system in potentiating the biologic, and possibly the clinical aggressiveness of these lymphoid tumors, and explore whether inhibition of the dysregulatory elements in this pathway can have therapeutic potential. We believe that our CD40 Signalosome model will provide an important "roadmap" into the important control mechanisms involved in NHL-B, that can be used for developing new therapeutic approaches for this very important group of human lymphoid neoplasms.

DESCRIPTION (provided by applicant): Mantle cell lymphoma (MCL) is the most therapy-resistant;B cell, non-Hodgkin's lymphoma (NHL-B) that currently lacks adequate therapy, and is increasing in incidence in the USA. Understanding the patho- physiology of MCL and experimental therapeutics for better MCL therapy has been hampered by the lack of a valid animal model. We have recently constructed the first transgenic MCL model that closely simulates the most therapy-resistant form of MCL, the Blastoid variant (MCL-BV). This model not only provides potential insights into the pathogenesis of MCL-BV, but also has a hierarchical tumor cell organization, with a small but distinct tumor cell subpopulation with self-renewal capabilities that resemble tumor initiating cells (TIC). The TIC population in our MCL-BV model also shows similarities to Cancer Stem Cells (CSC) that may represent the "cell of origin" in this MCL-BV model that should allow for testing various predictions of the Cancer Stem cell hypothesis, in a valid murine MCL model. Our MCL-BV model is a murine double transgenic (DTG) that was constructed by crossing IL14 x c-myc single TG mice. IL-14, a cytokine growth factor for normal B-lymphocytes, is over-expressed in NHL-B, where it may function as an oncogene in the B lymphocytic lineage, showing important B cell functional characteristics in addition to growth stimulation. DTG mice stochastically develop NHL-B resembling aggressive MCL-BV by 3-4 months (100% penetrance), allowing for sequential and defined patho-physiologic characterization of molecular and genotypic alterations in the murine B lymphoid cell compartment experimentally, from birth to lymphoma development. The DTG MCL-BV model provides multiple interesting potential insights into MCL, but our studies in this R21 proposal will focus on understanding one aspect in MCL-BV ontogeny, by identifying and characterizing a possible "cell of origin" in DTG/MCL-BV model. These CSC characteristics include tumor cell "self-renewal" characteristics and transplantabilty into immune-deficient (SCID) mice. In this proposal, we will focus primarily on: (1) characterizing the hierarchical nature of DTG tumor cell populations, identification of SP populations to support our hypothesis that our MCL model displays Tumor Initiating Cells (TIC) with Cancer Stem Cell (CSC) characteristics;and (2) further characterizing and validating that our DTG/MCL-BV model simulates anomalous cell cycle and constitutive growth/survival signaling pathway activation, characteristic of human MCL-BV. Our studies will explore how CSC-associated properties of putative MCL/ TIC in the DTG/MCL-BV model, contributes to the initiation, tumor maintenance, and possibly recurrence of the DTG lymphoma. We will also examine whether well-known human MCL characteristics, involving abnormal cell cycle regulation with cyclinD1 over-expression, and the constitutive activation of the NF-?B and pAKT growth and survival (anti-apoptotic) signaling pathways, are also activated in DTG/TIC tumor cells and/or other DTG tumor sub- populations, that may account for the clinical therapeutic resistance seen in patients with MCL, and particularly MCL-BV. PUBLIC HEALTH RELEVANCE: Mantle cell lymphoma (MCL) is the most difficult B cell Non-Hodgkin lymphoma to treat clinically, with the worst survival prognosis. Why MCL is so clinically aggressive is not known, which along with the lack of a suitable animal model has hampered development of more effective therapies. We have developed a genetically engineered model (double transgenic;DTG) of the blastoid variant of MCL (MCL-BV) that will provide both clues to the origin/causes of MCL as well as a model to test new MCL drugs, which will be developed in this grant proposal.

Project Summary Mantle cell lymphoma (MCL) has recently been recognized as a more complex NHL-B than the typical (small cell)/blastoid (large cell) dyad of the last two decades. Indolent, in situ, non-nodal MCL phenotypes indicate that MCL may represent a continuum from indolent lympho-proliferative disease (watch and wait) to the most aggressive disseminated blastoid MCL (hyper-CVAD). Validated patho-physiological and experimental therapeutic models of the MCL disease repertoire are serious unmet current needs for essential translational research in MCL. Based on our experimental studies, we are developing reproducible methodologies for generating both in vitro and in vivo MCL experimental models of MCL from patient samples. These models are based on developing extended growth and survival culture methods mediated by micro- environmental monocyte/macrophage MCL cell interactions. The signals that macrophages provide to induce MCL growth/survival and differentiation/transformation are completely unknown. In this proposal, we will utilize our novel in vitro MCL/macrophage co-culture systems and in vivo patient-derived xenotransplant MCL model systems to initially characterize and delineate the biologic functions of lymphoma- associated macrophages, and then elucidate potential pathways critical for the progression, dissemination and survival of MCL, as part of the MCL progression process from stable disease. This project is expected to provide novel insights into an unexplored area of MCL progression/transformation through microenvironment interactions that will not only fuel a better understanding of the disease process, but will facilitate the development of more mechanistic studies to identify novel druggable targets.