Steve Cole, Ph.D. - US grants

Development of a vaccine against HIV-1 is critical to containing the spread of AIDS in both the United States and the world. Research suggests that an effective HIV vaccine must induce strong cellular immune responses, but many current HIV vaccine preparations do not induce such responses. The proposed studies evaluate a novel strategy for enhancing cellular immune responses to HIV-1 vaccines using "molecular adjuvants" that regulate the activity of dendritic cells (DCs) as they initiate primary immune responses. In contrast to previous molecular adjuvants that encode growth factors or cytokines, the Transcription-Regulating Vectors (TRV) examined here express signaling molecules that simultaneously regulate the expression of a variety of DC genes involved in antigen presentation and the induction of cellular immunity. These studies evaluate the fundamental hypothesis that molecular adjuvants encoding pleiotropic transcriptional regulators can provide a more powerful and flexible means of enhancing vaccine- induced immune responses than do either traditional exogenous adjuvants or previous molecular adjuvants. The proposed studies address the following specific aims: Aim 1: Optimize transcription regulating vectors (TRVs) to enhance dendritic cell induction of primary cellular immune responses. Aim 2: Evaluate costimulation of dendritic cell NF-kappaB and PKA/Sp 1 pathways as a means of enhancing primary and secondary cellular immune responses to an HIV-1 vaccine. Aim 3: Evaluate the efficacy of TRVs in eliciting antiviral cellular immune responses in vivo using a mouse DNA vaccine model. These studies will establish a basic research foundation to support the subsequent application of TRV technology to construction of safe and effective antiviral vaccines suitable for worldwide use in stemming the spread of HIV-1 infection.

Antiretroviral medications currently represent the only effective means for controlling HIV pathogenesis, but existing drug regimens cannot fully stop viral replication in vivo. It is thus critical to identify physiologic processes that support residual HIV replication during highly active antiretroviral therapy (HAART). Recent data indicate that high levels of activity in the sympathetic division of the autonomic nervous system (ANS) represent an in vivo risk factor for elevated plasma viral load in untreated patients and those receiving HAART. The nervous system has long been known to regulate the activity of certain viral pathogens (e.g., herpes viruses), but little is known about its impact on lentiviruses such as HIV or SIV. ANS neurons innervate the lymphoid organs that serve as major sites of HIV-1 replication in vivo, and T lymphocytes bear beta adrenoreceptors that allow neural modulation of cellular activation, cytokine production, and cell trafficking. In previous studies, we found that the ANS neurotransmitter norepinephrine accelerates HIV-1 replication in vitro by altering cellular vulnerability to infection and enhancing viral gene expression. The proposed studies seek to define the molecular mechanisms of these effects. Specifically, these studies aim to: 1. Identify transcriptional mediators of ANS-induced HIV-1 gene expression. 2. Define ANS effects on soluble factors that modulate HIV-1 replication. 3. Assess interactions between ANS neurons and HIV/SIV replication in lymphoid tissues. These studies will establish a virologic framework for interventions aimed at blocking ANS support of residual HIV replication during HAART. Such interventions would enhance the efficacy of current antiretroviral treatment regimens by opposing physiologic processes that support ongoing pathogenesis.

DESCRIPTION (provided by applicant): Targeted therapy of cancer requires a clear understanding of the genetic alterations that drive malignant cell growth. Identification of causal genetic alterations is complicated by three characteristics of cancer etiology: 1.) multiple interacting alterations are often required to cause cancer, 2.) several distinct alterations may be sufficient to generate a single cancer phenotype, and 3.) oncogenic alterations appear in a dense background of normal genetic activity and spurious consequences of malignant cell growth. We propose to apply a variant of the machine learning algorithm PRIM to the task of identifying disjunctive sets of conjunctive genetic alterations that cause specific cancers or provide prognostic information about clinical course and treatment efficacy. These analyses synthesize information from low-level bioinformatics resources we have already developed to map chromosomal alterations and monitor global patterns of transcription factor activity. Based on those foundations, the present studies develop high-level analytic tools to map combinatorial interactions among low-level genomic events. Specifically, these studies seek to: Aim 1: Develop graphical user interface (GUI) software to support combinatorial genomic analyses by biologists with limited computational background. Aim 2: Optimize combinatorial prediction of disease progression and treatment response. Aim 3: Develop PRIM-based statistical models to identify functional complementation groups of genetic alterations and transcriptional control signals. The bioinformatic tools produced in these studies will create a generalized analytic infrastructure for mapping complex etiologies in cancer and deploying patient-specific targeted therapies.

DESCRIPTION (provided by applicant): The proposed research seeks to determine how adverse social environments influence the risk of inflammation-related disease by up-regulating the expression of pro-inflammatory genes. These studies test the hypothesis that adverse social environments stimulate the hematopoietic production of immature pro-inflammatory monocytes (CD16- in humans, Ly-6c-high in mice) via threat-induced activation of beta-adrenergic receptors in bone marrow myelopoietic cells. Specific aims will: (Aim 1) Define the neural and endocrine pathways by which chronic threat up-regulates pro-inflammatory monocytes; (Aim 2) Define the specific beta-adrenergic receptors and target cell types mediating threat-induced expansion of pro- inflammatory monocytes; and (Aim 3) Define the myelopoietic molecules mediating beta-adrenergic expansion of pro-inflammatory monocytes (including GM-CSF, TGF-beta, and the CXCL12/CXCR4 chemokine signaling axis). When complete, these studies will provide an integrated mechanistic model of the neural / hematopoietic pathway by which chronic adversity can up-regulate inflammatory gene expression in circulating immune cells. The overarching goal of these studies is to develop a comprehensive theory that explains how common social risk factors can influence multiple inflammation-related diseases. In addition to clarifying the basic physiologic mechanisms involved in defensive programming of the immune system transcriptome, these studies will identify specific CNS mechanisms (e.g., Crf gene activation in central nucleus of the amygdala), pharmacologic intervention strategies (e.g., beta-2 and beta-3 adrenergic antagonists, and antagonists of GM-CSF, TGF-beta, and/or CXCR4), and mechanistic biomarkers (e.g., myelopoietic molecules and circulating monocyte phenotypes) that can be applied in future studies to clarify how stress-induced up- regulation of pro-inflammatory monocytes impacts specific inflammation-related diseases such as atherosclerosis, Type II diabetes, Alzheimer's disease, and cancer.

Overview Abstract The USC/UCLA Center on Biodemography and Population Health (CBPH) represents a unique and highly successful collaboration between the Davis School of Gerontology of the University of Southern California (USC) and the Multi-campus Program in Geriatric Medicine and Gerontology in the Geffen School of Medicine at the University of California at Los Angeles (UCLA), each of which focuses exclusively on research and teaching on aging. The CBPH has a longstanding role as a leader in efforts to promote theory-based integration of biological measurement into population-based studies, on-going development and validation of biological measurement protocols, and theoretically motivated research on the biological mechanisms by which social, economic, psychological, medical and environmental factors ?get under the skin? to influence the process of health change with age. The CBPH has developed unique clinical and laboratory infrastructure and pilot projects to improve understanding and use of biodemographic indicators, increase indicators available to population studies, support more reliable and valid collection of data across a large number of national and international surveys, and made advances in measurement and validation that allow population surveys to keep pace with scientific advances in the science of aging. This application proposes a set of activities designed to (i) expand and enhance theoretical development of the field of biodemography so that we focus on a new generation of biomarkers reflecting molecular and cellular processes that reflect the basic mechanisms of aging, (ii) continue efforts to attract new and promising researchers to the field, and (iii) enhance our Center?s unique role in supporting development, validation, implementation and dissemination of new and better biodemographic measurement protocols. The specific aims of the CBPH will be to: (1) support and foster biodemographic research to understand the multiple and interacting factors that affect population health (with a particular focus on expanding and deepening our understanding of the biological pathways through which experiences and exposures over the life-course impact trajectories of health and how such influences may vary across subgroups and settings); (2) further develop an active biodemographic research community by engaging established and promising junior researchers in a network of scholars who can help advance biodemographic research; (3) offer funding for pilot projects to support cutting-edge biodemographic research; (4) support development and dissemination of new research technologies, methodologies and data through our Research Resource & Dissemination Core and dissemination activities of Administrative Core. Such advances are needed to support development of models of population health that provide much needed evidence for policy planners regarding the most effective points of intervention to improve population health and reduce health disparities. The CBPH has developed the infrastructure, resources and expertise to accomplish its goals and continue its significant leadership role in advancing biodemographic research on aging. OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015) Page Continuation Format Page