|
|
||||
|
|
|
|
|
 |
 |
 |
 |
 |
 |
 |
 |
|
|
University of California, Los Angeles
Project 1 This proposal is built upon an ongoing population-based case-control study on marijuana use and the risk of lung and UAT cancers funded by NIH (R01 DA/CA 11386). The objectives are to better understand the molecular mechanisms for the risk of lung cancer. We propose that the development of lung cancer is an interactive process involving previous environmental exposures such as active and passive tobacco smoking, marijuana smoking; intrinsic host susceptibility such as polymorphisms and methylations of the GST P1 and polymorphisms of the TP53 gene and genetic instability including TP53 and mutations and p16 methylations and other alterations. The study will be based on 600 cases and 600 controls interviewed by the parent study. Among those, we project 420 lung cancer patients with have their tumor tissue available for the analysis and 488 case and 488 controls will have their buccal cell samples for this proposed study, according to our pilot study. Epidemiological factors will be obtained by face-to-face interview. Our study is designed to fulfill these specific aims. (1) We will assay polymorphisms and hypermethylations of GST P1 in 488 cases and 488 controls, to evaluate the effects of those alterations on the risk of lung cancer, and explore gene-environment interaction between GSTP1 and environmental exposures. Other metabolic genes involved in PAH metabolisms such as GST M1and P4501A1 will be assayed. (2) We will measure polymorphisms of TP53 gene in 488 cases and 488 controls and examine the effects of polymorphisms of TP53 gene by a case-control study. We will measure TP53 mutations by PCR-SSCP and sequencing to test the hypothesis that cases with lung cancer with and without p53 mutations are etiologically distinctive groups with regard to major risk factors such as active and passive tobacco smoking, marijuana smoking, occupational exposure, etc. (3) We will measure p16 methylations and other alterations including mutations, homozygous deletions, and microsatellite instability, and correlate these alterations with tobacco smoking and occupational exposures. The result of this study may provide insight into lung carcinogenesis. It may assist us to identify high-risk individuals for intervention and may have translational potential to screening, early detection and prognostic prediction.
Project 2 Lung cancer is associated with an overall 15% survival and is the most common cause of cancer death in the United States. Helical computed tomography (hCT) is very sensitive for detecting lung cancer by virtue of its cross-sectional perspective and ability to acquire volumetric data sets of the chest in single sequences of high resolution. Preliminary data show that lung cancers detected with hCT are frequently small, early stage lesions; however, indeterminate nodules are observed in 25% to 50 % of screened individuals, the vast majority of which will be benign. The exclusion of malignancy may require biopsy, surgery, or prolonged follow-up with CT. There is a compelling need to develop methods of accurate, non-invasive nodule characterization, possibly by providing in vivo surrogates of the aberrant angiogenesis and altered cellular metabolism that are basic to neoplastic proliferation. Both contrast-enhanced CT and positron emission tomography (PET) are able to discriminate benign from malignant lung nodules. The former, based upon the principle that tumors enhance because of increased vascularity, has a high negative predictive value in lesions 7 mm or greater, but is relatively nonspecific, while PET using 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) is accurate, but reliable primarily for larger lesions of at least 15 mm diameter. We propose to develop a combined CT/PET approach for nodule characterization, using image analysis with feature classification to investigate several potentially complementary imaging features to better discriminate benign and malignant lesions. From hCT, basic 2-D non-visual texture features; 3-D volume, shape, and surface features; and post-contrast enhancement patterns will be input to linear discriminant and non-linear neural net classifiers to determine the panel of features that provides the greatest accuracy. Semi-quantitative measures of metabolic activity in lung lesions will be calculated from FDG PET scans that have been optimally corrected for photon attenuation and partial volume effects by subject-specific anatomical templates created from hCT. The diagnostic performance of imaging features will be determined individually and in combination relative to diagnoses confirmed pathologically or by stable radiographic appearance over two or more years. In addition to predicting malignancy in vivo, the relationships between CT and PET features to morphometric and immunohistochemical preparations of lesions will be analyzed with a goal to identifying imaging patterns that are more predictive of tumor biology and clinical outcomes than is currently possible with classical descriptions of tumor size and stage. Our hypothesis is that the information derived from x-ray CT and FDG-PET can combine synergistically to improve the accuracy of nodule characterization for lesions both within and below the current threshold of accuracy of either modality.
Project 3
Neuropeptides, including mammalian bombesin-like peptides gastrin and cholecystokinin, are a structurally diverse group of molecular messengers that function in a rich network of intercellular communication. These signaling peptides have been identified in our laboratory as potent cellular growth factors. Neuropeptides and their receptors are a major driving force behind one of the most clinically aggressive cancers, small cell lung cancer (SCLC) . Our central hypothesis is that the surface receptors and the intracellular signaling pathways that mediate the proliferative and migratory responses induced by neuropeptides are potential targets for novel therapeutic interventions. Given the pivotal role of the protein kinase C (PKC) family in neuropeptide signal transduction, the studies proposed under specific aim 1 will focus on the novel PKC/PKD phosphorylation cascade in SCLC cells.
Project 4 Our efforts to produce an effective cancer therapy are focused on methods to restore tumor antigen presentation. Dendritic cells (DC) are bone marrow-derived leukocytes characterized by the high level expression of major histocompatibility complex (MHC) and co-stimulatory molecules as well as the capacity to take-up, process and present antigens. These powerful capacities facilitate activation and expansion of antigen-specific T cells. Recent murine studies and clinical trials have shown that DC, when appropriately armed with a tumor antigen (Ag), can promote antitumor immunity and significant tumor regression. In this proposal we hypothesize that autologous DC, when transduced with an adenoviral vector expressing the IL-7 gene (DC-AdIL-7), can be used to stimulate specific and therapeutic anti-tumor immunity without the need for priming with a tumor antigen ex vivo. In preliminary studies, the intratumoral injection of these gene-modified DC into established murine tumors induced specific antitumor responses both locally and at metastatic sites. Animals treated in this manner not only experienced complete tumor regression, but were protected from subsequent tumor challenge. We hypothesize that the autologous tumor provides access to the entire repertoire of available antigens, both increasing the likelihood of a response and reducing the potential for phenotypic modulation. The overall goal of this proposal is to use murine models to determine the immunologic mechanisms by which DC-AdIL-7 mediate tumor eradication. The specific aims are: 1) to identify the mechanisms of antitumor responses in DC-AdIL-7 intratumoral therapy and 2) to determine the features of transferred DC responsible for mediating an effective antitumor response. A unique focus of this work is the emphasis on a DC-based approach to stimulate specific immune responses that does not exclude patients on the basis of HLA phenotype or because of lack of expression of a particular tumor antigen. Thus, this therapy would be available to all lung cancer patients in the appropriate clinical setting. We anticipate that the studies described in the current proposal will enhance our understanding of the complex interactions between tumor cells and DC and thus lead to more effective therapy for lung cancer. Based on the preliminary findings in this study the Clinical Trials Core has designed a Phase I/II protocol that will evaluate the intratumoral injection of IL-7-modified DC in endobronchial tumors in patients with IIIb and IV NSCLC.
Project 5 Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related mortality in the US. Clearly, new strategies for therapeutic intervention are necessary. While carcinogenesis is related to genetic and epigenetic events, NSCLC growth is dependent upon angiogenesis. Net tumor-derived angiogenesis is due to an imbalance in the over-expression of angiogenic, as compared to angiostatic factors. This aberrant angiogenesis allows for the perpetuation of tumor growth and eventual metastasis. However, the mediators that orchestrate this aberrant neovascular response in NSCLC have not been fully elucidated. Members of the CXC chemokine family exert disparate function in regulating angiogenesis related to three amino acid residues (the 'ELR' motif) in the NH2-terminus of these cytokines. CXC chemokines have potent angiogenic (ELR+) and angiostatic (ELR-) activity. Our central hypothesis is that net angiogenesis during tumorigenesis of NSCLC is determined, in part, by an imbalance in the expression/function of CXC chemokines that favors the over-expression of angiogenic, as compared to angiostatic CXC chemokines. This paradigm predicts an environment that perpetuates an angiogenic phenotype of the endothelium promoting tumorigenesis, and metastases. The proposed studies will focus on the following specific aims: I) To demonstrate that vascular endothelial growth factor (VEGF)-mediated angiogenesis is related to enhanced endothelial cell survival by Bcl-2- and interleukin-8-dependent mechanism(s). II) To establish that the CXC chemokine receptor, CXCR2, is the putative receptor for IL-8/ELR+ CXC chemokine mediated angiogenesis. III) To determine the mechanism(s) for angiostatic interferon (IFN)-inducible (ELR-) CXC chemokine inhibition of VEGF-, bFGF-, EGF-, ELR+ CXC chemokine-induced angiogenesis. IV) To demonstrate in vivo that the IFN-inducible angiostatic ELR- CXC chemokines account for the angiostatic effects of IL-18, IL-12 and IFN-g in mediating inhibition of tumor-derived angiogenesis, tumorigenesis, and spontaneous metastases. V) To establish that human specimens of NSCLC demonstrate an imbalance in the expression of angiogenic ELR+, as compared to angiostatic IFN-inducible ELR- CXC chemokines, and this imbalance is directly correlated to tumor-derived angiogenesis and recurrence of surgical stage I and II NSCLC. These experiments will use molecular, cellular, whole animal models, and human specimens to assess angiogenesis related to NSCLC tumorigenesis. The experiments designed in this proposal will demonstrate that CXC chemokines mechanistically play critical roles in regulating angiogenesis during NSCLC tumorigenesis. These experiments will provide the foundation for the development of novel therapeutic strategies to modulate this biology and attenuate tumor-derived angiogenesis, that ultimately will reduce NSCLC tumorigenesis and metastases.
Core 1 This Clinical Trials Core is established to identify relevant translational research, work with SPORE investigators to formulate hypothesis driven clinical investigations based on their discoveries, and provide the personnel/resources needed to support exploratory phase I/II clinical trials in patients with lung cancer. Research with a potential for clinical trial applications will be identified within the organizational structure of the SPORE. Robert A. Figlin, M.D., Director of Clinical Operations for the Core, will serve on the SPORE Executive Committee and Co-Chair the Developmental Programs Selection Committee, to assure that applicable projects are identified and prioritized. Similarly, Michael D. Roth, M.D., Director of Research Operations for the Core, will Co-Chair the Career Development Selection Committee. Once projects with clinical potential are identified, the Core Directors will meet with responsible investigators to collaborate in the formulation of testable clinical hypotheses. To implement clinical trials, a Core Administration and Support Staff will be developed, building upon existing resources available through the UCLA Thoracic Oncology and the Lung Cancer Research Programs. Supporting resources required to carry out clinical trials work will be obtained by interacting with the following established programs: 1) Thoracic Oncology Program and Jonsson Comprehensive Cancer Center's Clinical Research Unit; 2) Lung Cancer Research Program; 3) NIH-supported General Clinical Research Center; 4) Adoptive Immunotherapy Cell Culture Facility; 5) Pathology Human Tissue Research Center and Core; 6) Biostatistics Core; 7) UCLA Research and Drug Information Pharmacy; 8) Thoracic Imaging Division of Radiologic Sciences; 9) Thoracic Oncology Tumor Board; 10) Internal Scientific Peer Review Committee; 11) UCLA Institutional Review Board; 12) JCCC Quality Assurance Program; and 13) the established clinical programs, clinics and participating physicians from the Pulmonary & Critical Care Medicine, Hematology/Oncology, Thoracic Surgery and Radiation Oncology services within the Greater UCLA HealthCare System. Once projects have been formulated into working Clinical Protocols, funds will either be budgeted directly from the UCLA Lung Cancer SPORE to carry out exploratory phase I/II clinical trials or independent grants will be submitted for supplemental clinical trial support. Working in conjunction with the research staff from Project IV, the Clinical Trials Core has developed, and will carry out, an initial clinical trial entitled, "A Phase I/II Trial Evaluating Intratumoral Injection of Interleukin-7 Gene-Modified Autologous Dendritic Cells for the Treatment of Non-Small Cell Lung Cancer".
Core 2 Numerous aspects of lung cancer research are dependent on pathology diagnostic expertise as well as numerous pathology-related services. The Pathology Core Resource is dedicated to providing pathology support services for investigators of the UCLA Lung SPORE program. These services include 1) human and veterinary pathology consultation, 2) procurement of remnant tissue samples, 3) histology, 4) immunohistochemistry and in situ hybridization, 5) laser-capture microdissection, 5) electron microscopy, 6) morphometry and imaging, 7) tissue arrays, and 8) molecular pathology. Notably, the Pathology Core Resource is currently functional and highly successful. Services and technologies provided by the Pathology Core Resource are critically important for understanding the pathogenesis of diseases such as lung cancer. As such, all projects presented in this grant propose to utilize one or more components of this shared resource. In addition to the scientific advantages that this core affords, this resource is cost-effective as the cost for individual researchers to perform such activities on their own would be prohibitively expensive. The Pathology Core Resource is directed by a team of faculty members with expertise in the specific pathology services mentioned as well as experience in tumor biology research. In addition, the technical staff is highly skilled. Significantly, all members of the Pathology Core Resource are committed to the success of the UCLA Lung SPORE program. Thus, our Aim here is to supplement the funding of this facility and therefore: i) enable continued expansion and optimization of services; and ii) allow a focused effort to provide services specifically dedicated to promoting lung cancer research at UCLA.
Core 3 The Biostatistics Core is designed as an extension of the UCLA Jonsson Comprehensive Cancer Center B.A.S.E. Unit (Biostatistics, Analytic Support and Epidemiology Shared Resource), a NIH and institutionally-funded resource providing complete biostatistical and biomathematical support for Cancer Center investigators in the areas of basic science and clinical research design, data analysis, modeling, computing and data management. Under the direction of Robert Elashoff, Ph.D., and with dedicated support for data management, the SPORE Biostatistics Core will interact with the B.A.S.E. Unit and with all SPORE Investigators to provide comprehensive biostatistical consulting and analysis. Additional input from Biostatistics and Biomathematics faculty with expertise in statistical genetics, pharmacokinetics and pharmacodynamics, multivariate methodology, categorical data, survival analysis, clinical trials and translational research, regression methods, imaging and functional analysis will be available.
Core 4 The Administrative Core will be responsible for the overall functioning of the SPORE programs. The purpose of the Administrative Core is to provide scientific leadership and administrative support for all aspects of the SPORE program. This will include grants and financial management, scheduling of meetings and seminars, assistance with manuscript preparation and coordination of activities with the Cancer Center and all other UCLA programmatic areas. The CORE will be responsible for coordinating the filing of progress reports and for communications with the NCI as well as other outside agencies. Together with the Clinical Trials Core, the Administrative Core will assure the necessary completion of documents for regulating agencies. The Administrative Core will be responsible for scheduling the Executive Committee, the Career Development Selection Committee, and the Developmental Program Selection Committee meetings. In addition, the Administrative Core will arrange the meetings of the Internal and External Advisory Boards. The organization, scheduling, and advertising of the Annual UCLA Lung Cancer SPORE Symposium will be the responsibility of the Core. The Administrative Core will be responsible for the administering the Developmental Research and Career Developmental Programs. This will include the oversight of established policies for the recruitment of women and minorities. Drs. Dubinett and Figlin have worked closely on administrative matters related to lung cancer investigations for several years. As the Leader and Co-Leader of this Core, they provide the necessary scientific and administrative leadership. As leaders of the administrative core they will assure that all core functions are fulfilled.
List of Investigators |
 |
|||
|
|