Sonia C. Flores - US grants

The aims of this project are to correlate changes in SOD levels with a particular disease, namely ARDS and to assess the effectiveness of the SOD variants at alleviating the damage in a rat model of acute lung injury. This project will also address the mechanisms of extracellular superoxide dismutase alterations in mammalian tissue culture models of oxidative stress and identify possible markers of acute lung injury in vivo.

DESCRIPTION (Adapted from applicant's abstract) Background: Myocardial inflammation with pericardial effusions leading to cardiac tamponade and cardiomyopathies are a frequent finding in AIDS patients. Up-regulation of microvascular endothelial cell adhesion molecule (CAM) expression by oxidants and/or circulating cytokines produces a hyperadhesive endothelium that preferentially sequesters leukocytes and contributes to the inflammatory process. Further secretion of oxidants, pro-inflammatory mediators and/or proteases by these cells causes alterations to the endothelial permeability barrier, setting up a cascade of events that end in cardiomyocyte dysfunction and cardiac failure. The cardiac pathologies in AIDS patients occur even in the absence of known opportunistic infections or direct infection by HIV, suggesting the existence of a soluble factor, either of viral of cellular origin. This soluble factor may be the HIV-1 Tat protein. It is proposed that release of the soluble HIV-1 Tat regulatory protein from infected cells and its uptake by uninfected cells increases oxidant stress, affects NF-kB activation and elevates CAM levels, which sequesters leukocytes and depresses cardiac cell function. The mechanisms underlying this process relates to inhibition of the target cell's antioxidant enzymes Mn-SOD and is reflected by a perturbed GSH/GSSG ratio. Therefore, the specific aims are to test the hypotheses that 1) Tat-dependent Mn-SOD inhibition of cardiac microvascular endothelial cells increases oxidant stress 2) that Tat-mediated oxidants stress results in CAM up-regulation, possibly via NF-kB and/or AP-1 activation 3) that Tat increases cytokine secretion 4) that these Tat-mediated alterations increase leukocyte-endothelial cell interactions 5) that Tat also mediates increases in vascular permeability. All of these events converge to affect the integrity of cardiac endothelium and may therefore result in some of the cardiovascular abnormalities seen in AIDS. The effect of Tat on cardiac endothelial cell, and/or leukocyte function will be evaluated in cells isolated from human sources and from Tat-transgenic mice. The proposed research addresses one of the important questions regarding the etiology of cardiovascular complications in AIDS: why is there evidence of myocarditis, yet very little evidence of a pathogenic viral, bacterial, or fungal agent? Tat-mediated oxidative stress may be an important contributor to the cardiac inflammation seen in these patients. (End of Abstract)

DESCRIPTION (Adapted from applicant's abstract) Background: Myocardial inflammation with pericardial effusions leading to cardiac tamponade and cardiomyopathies are a frequent finding in AIDS patients. Up-regulation of microvascular endothelial cell adhesion molecule (CAM) expression by oxidants and/or circulating cytokines produces a hyperadhesive endothelium that preferentially sequesters leukocytes and contributes to the inflammatory process. Further secretion of oxidants, pro-inflammatory mediators and/or proteases by these cells causes alterations to the endothelial permeability barrier, setting up a cascade of events that end in cardiomyocyte dysfunction and cardiac failure. The cardiac pathologies in AIDS patients occur even in the absence of known opportunistic infections or direct infection by HIV, suggesting the existence of a soluble factor, either of viral of cellular origin. This soluble factor may be the HIV-1 Tat protein. It is proposed that release of the soluble HIV-1 Tat regulatory protein from infected cells and its uptake by uninfected cells increases oxidant stress, affects NF-kB activation and elevates CAM levels, which sequesters leukocytes and depresses cardiac cell function. The mechanisms underlying this process relates to inhibition of the target cell's antioxidant enzymes Mn-SOD and is reflected by a perturbed GSH/GSSG ratio. Therefore, the specific aims are to test the hypotheses that 1) Tat-dependent Mn-SOD inhibition of cardiac microvascular endothelial cells increases oxidant stress 2) that Tat-mediated oxidants stress results in CAM up-regulation, possibly via NF-kB and/or AP-1 activation 3) that Tat increases cytokine secretion 4) that these Tat-mediated alterations increase leukocyte-endothelial cell interactions 5) that Tat also mediates increases in vascular permeability. All of these events converge to affect the integrity of cardiac endothelium and may therefore result in some of the cardiovascular abnormalities seen in AIDS. The effect of Tat on cardiac endothelial cell, and/or leukocyte function will be evaluated in cells isolated from human sources and from Tat-transgenic mice. The proposed research addresses one of the important questions regarding the etiology of cardiovascular complications in AIDS: why is there evidence of myocarditis, yet very little evidence of a pathogenic viral, bacterial, or fungal agent? Tat-mediated oxidative stress may be an important contributor to the cardiac inflammation seen in these patients. (End of Abstract)

[unreadable] DESCRIPTION (provided by applicant): Human immunodeficiency virus infection results in significant organ injury due to inflammation. The lung and the heart are particularly prone to develop inflammation-mediated damage, thus resulting in cardiopulmonary morbidity and mortality. In recent years, it has become apparent that reactive oxygen and nitrogen species play a fundamental role in the initiation, amplification and cell destruction of innate and acquired inflammatory processes. The Tat protein of HIV, a viral regulator required for efficient transcription of the HIV genome during host infection, also activates the transcription of NF-K-B-dependent key inflammatory molecules. We demonstrated that Tat induces oxidative stress and partial depletion of glutathione. This effect is partly mediated by Tat-dependent inhibition of Mn-superoxide dismutase (MnSOD) expression, resulting in increased levels of oxidants. We have also demonstrated that one of the downstream effects of Tat-mediated oxidative stress is the activation of endothelial cell NF-KB, which is a potent inducer of proinflammatory molecules such as the leukocyte adhesion molecule ELAM and the cytokines TNF-a, IL-1 and IL-6. The enhanced oxidative stress in patients with HIV infection may result in direct tissue damage. We hypothesize that HIV-1 Tat protein represses antioxidant systems, resulting in increased free radical species generation and enhanced activation of NF-kB-dependent pro-inflammatory mediators. Lung Tat expression leads to chronic inflammatory injury per se and amplifies lung tissue destruction in the setting of increased oxidative stress due to environmental stimuli or in pro-inflammatory states induced by bacterial products. Abrogation of Tat-induced oxidative stress may lead to significant lung protection against HIV-induced lung inflammation. To address this hypothesis, we have engineered several transgenic (Tg) mouse lines with either low, intermediate or high lung-specific expression of the Tat protein. Preliminary evidence suggests these mice are oxidatively stressed and are sensitized to pro-inflammatory agents. Using this animal model of Tat-mediated pulmonary oxidative stress, we propose to address the following questions: 1) Does expression of the HIV-1 Tat increase basal oxidant stress in lungs? 2) Does Tat-mediated oxidative stress increase basal NF-&-dependent pro- inflammatory mediators and amplify inflammatory damage? 3) Does antioxidant treatment reduce inflammation induced by Tat alone or that induced by the combination of oxidative stress and Tat? This animal model will provide important insight on how retroviral products reset the OxidanVantioxidant balance and thus predispose to or cause inflammatory cell-mediated organ injury and disease. Furthermore, these studies should provide important information regarding the rationale for an antioxidant regime to treat HIV-induced organ dysfunction.

DESCRIPTION (provided by applicant): The University of Colorado Health Sciences Center (UCHSC) and its Graduate School seek renewal of NHLBI funding for a minority summer Biomedical Sciences Research Internship Program (BioRIP). The current grant was first funded on June 2000 and provided funds for 6 trainees the first 2 years and 8 in subsequent years. The goal of this program has been to provide a stimulating research experience and recruit these students to attend graduate or professional schools and continue studies that will lead to a career in biomedical research. The UCHSC has an established commitment to increasing the diversity of its student population and to provide an experience in contemporary biomedical research to undergraduates that are members of under-represented populations. In addition to the NIH-funded slots, the President's Office of the University of Colorado and the Chancellor of UCHSC believe that the student population should be as diverse as possible and have therefore provided combined funds for 2-3 additional trainees. These trainees would not normally qualify as underrepresented minorities but are economically disadvantaged, from rural areas or first generation college attendees. In addition, funds have also been provided for training of talented minority high school students. The combined program, called Graduate Experiences for Multicultural Students (GEMS), has funded a total of 42 students in the past 4 years. The majority (88%) of these students have expressed a commitment to continue professional studies. Of the 26 students who have already graduated, 9 are enrolled in professional schools, and 13 are in the process of applying to graduate science and professional programs. All of the students who are still in school will definitely pursue post-graduate degrees in science or medicine. The current application builds upon this success and is seeking to increase the number of NIH-funded student slots to 10 and continue offering mentored research experiences in one of our research laboratories. These laboratory rotations will be coupled with an introductory course on molecular biological and biochemical techniques and overviews of biochemistry and cell biology. In addition, the students will participate in brown bag lunches covering topics such as "Ethics in Science and Medicine", How to write a research paper", or "How to put together an effective oral presentation". The UCHSC is unique in that there are no undergraduates at our institution, and that the majority of our research focuses upon health-related issues. Many of the undergraduates we have trained never had contact with a medical school, even if one exists on their campus, and therefore the experience on our campus represents the first contact they have with biomedical research in a dedicated setting. The University of Colorado Health Sciences Center and its neighboring institution, the National Jewish Medical and Research Center, offer a very talented group of investigators with a diverse interest in other biological problems of significance to the health sciences. These institutions offer a particular strength in pulmonary diseases. The research and educational components of BioRIP is also complemented by personal advising on health professional careers, and gaining admission to professional schools.

DESCRIPTION (provided by applicant): Primary pulmonary hypertension (PPH), a rapidly progressive and usually fatal disease, has an incidence rate among the HIV infected population many times higher than in the general population. Unfortunately, the pathogenesis of HIV-related pulmonary hypertension (HRPH) is not well understood. Nevertheless, the histological similarities are striking: uncontrolled endothelial cell (EC) proliferation and formation of plexogenic lesions obliterate the Lumina of the pulmonary arteries with subsequent right heart failure. The immune dysregulation, chronic exposure to viral products in the lung and altered chemokine/cytokine profile may contribute to the injury. In the lung, HIV-1 infects primarily macrophages providing a potential reservoir of virus and a source of localized viral proteins such as Nef, which can circulate and affect surrounding cells. Studies of HRPH have been hampered by lack of a suitable animal model. Since numerous primate models of HIV-1 recapitulate the immune deficiencies and complications seen in humans, we undertook a study of lungs from macaques infected with an SIV/HIV chimeric virus containing HIV-1 Nef (SHIVnefSF33A) and found plexogenic lesions in the lungs of SHIV-nef but not in SIV Nef-infected macaques, suggesting that there are functional differences between the nef alleles in their ability to promote pulmonary vascular remodeling. We propose to study the natural history and progression of HRPH in SHIVnef infected monkeys. Our specific hypothesis is that immune dysregulation of SHIVnef-infected monkeys, triggers a phenotypic switch in EC that allows selection of a highly proliferative, growth-dysregulated EC population that obliterates the Lumina of pulmonary arteries through plexiform lesion formation. To study this, we will address the following question: What is the natural history of PH in macaques infected with SHIVnef, and in a background of gammaherpesvirus infection? We will infect the monkeys, track PH development post-infection, and will correlate immunological parameters with lesion formation. Does HIV nef lead to the acquisition of a proliferative phenotype in lung microvascular EC? We will examine the in vitro proliferative properties of pulmonary endothelial cells after exposure to various nef alleles/mutants or to conditioned media from macrophages exposed to these as well. Using a primate model system that is phylogenetically very close to humans allows us to study both the initiation and progression phases of HRPH.

DESCRIPTION (provided by applicant): The University of Colorado Health Sciences Center (UCHSC) and its Graduate School seek renewal of NHLBI funding for a minority summer Biomedical Sciences Research Internship Program (BioRIP). The current grant was first funded on June 2000 and provided funds for 6 trainees the first 2 years and 8 in subsequent years. The goal of this program has been to provide a stimulating research experience and recruit these students to attend graduate or professional schools and continue studies that will lead to a career in biomedical research. The UCHSC has an established commitment to increasing the diversity of its student population and to provide an experience in contemporary biomedical research to undergraduates that are members of under-represented populations. In addition to the NIH-funded slots, the President's Office of the University of Colorado and the Chancellor of UCHSC believe that the student population should be as diverse as possible and have therefore provided combined funds for 2-3 additional trainees. These trainees would not normally qualify as underrepresented minorities but are economically disadvantaged, from rural areas or first generation college attendees. In addition, funds have also been provided for training of talented minority high school students. The combined program, called Graduate Experiences for Multicultural Students (GEMS), has funded a total of 42 students in the past 4 years. The majority (88%) of these students have expressed a commitment to continue professional studies. Of the 26 students who have already graduated, 9 are enrolled in professional schools, and 13 are in the process of applying to graduate science and professional programs. All of the students who are still in school will definitely pursue post-graduate degrees in science or medicine. The current application builds upon this success and is seeking to increase the number of NIH-funded student slots to 10 and continue offering mentored research experiences in one of our research laboratories. These laboratory rotations will be coupled with an introductory course on molecular biological and biochemical techniques and overviews of biochemistry and cell biology. In addition, the students will participate in brown bag lunches covering topics such as "Ethics in Science and Medicine", How to write a research paper", or "How to put together an effective oral presentation". The UCHSC is unique in that there are no undergraduates at our institution, and that the majority of our research focuses upon health-related issues. Many of the undergraduates we have trained never had contact with a medical school, even if one exists on their campus, and therefore the experience on our campus represents the first contact they have with biomedical research in a dedicated setting. The University of Colorado Health Sciences Center and its neighboring institution, the National Jewish Medical and Research Center, offer a very talented group of investigators with a diverse interest in other biological problems of significance to the health sciences. These institutions offer a particular strength in pulmonary diseases. The research and educational components of BioRIP is also complemented by personal advising on health professional careers, and gaining admission to professional schools.

DESCRIPTION (provided by applicant): Little is known about the pathogenesis of pulmonary arterial hypertension (PAH) and the factors that contribute to its higher prevalence in HlV-infected patients. Chronic exposure to viral products in the lung as well as HlV-induced immune dysregulation may contribute to pulmonary vascular disease. The experiments proposed in this grant application are in response to the RFA-HL-07-008 titled "Longitudinal Studies of HIV- Associated Lung Infections and Complications''and will expand our initial studies that found an association between the HIV-1 Nef (negative factor) protein and HIV-related pulmonary arterial hypertension (HRPAH). In collaboration with Dr. Marc Humbert from France, we obtained peripheral blood mononuclear cell DNA (PBMC) from French HRPAH patients and sequenced nef alleles in these samples. Phylogenetic analyses revealed that the majority of sequences clustered with subtype B sequences. We therefore compared the French Nef sequences with all HIV-1 subtype B sequences in published databases. We found that particular nef molecular signatures occurred more frequently in French HRPAH patients than in the databases. We therefore hypothesize that allelic variants of nef, which can be identified by unique molecular signatures, are associated with the development of pulmonary arterial hypertension (PAH) in patients already diagnosed and predict disease progression in a population of patients at risk as assessed by echocardiography. These variants may trigger a cascade of events in the lungs that allows for the selection of a rapidly growing, apoptosis-resistant endothelial cell population. We will rely upon two distinct, well-established cohorts of HIV-1-infected subjects - a group of French patients with HRPAH diagnosed by right heart catheterization (RHC), the diagnostic gold standard, and a cohort of HIV-1- infected individuals in San Francisco who have been diagnosed with elevated pulmonary artery systolic pressures (PASP) by echocardiography and who may be at a high mortality risk. Retrospective and prospective plasma, PBMC and bronchoalveolar lavage fluid cells (BALF) from the two clinical sites will be sent to the research site in Denver where nef sequences will be obtained, analyzed and characterized. We aim to evaluate the amino acid sequence and known domain structures from patients with and without HRPAH to determine if common sequence disruptions or similarities are associated with disease pathogenesis;to evaluate whether nef alleles from HIV-1 infected patients with elevated PASP and at risk of developing PAH in San Francisco have signature sequences similar to sequences identified in French patients and that may predict progression to HRPAH;and to identify and compare nef sequences isolated from BALF with sequences found in PBMC and plasma in patients with and without elevated PASP in San Francisco for evidence of compartmentalization. Finally, we will evaluate whether nef alleles identified in these patients induce proliferation in cultured human lung microvascular endothelial cells. For all cohorts, DNA, RNA, plasma, recombinant vectors, BALF cell samples, immunophenotypes and data as well as molecular tools generated from these studies will be deposited with the Data Coordinating Center (DCC) and made available for future mechanistic studies.

DESCRIPTION (provided by applicant): In 2004 the University of Colorado Health Sciences Center consolidated its campus with the University of Colorado at Denver downtown campus to become the University of Colorado at Denver Health Sciences Center (UCDHSC). Because urban high schools with sizable populations of ethnic and racial minorities have to contend with a myriad of cultural and societal issues affecting basic achievement and academic proficiency, students with underlying talent and capability are too often not afforded the intellectual support available to students from more affluent suburban school districts. We will intervene by providing promising students with a program designed to strengthen their fundamental skills, enhance their academic credentials, and elicit their passion for science. Our objectives are (1) to provide a Summer Science Academy designed to improve any deficiencies in basic math, science, or language skills;(2) to use supplemental instruction to improve retention and enable at least 80% of participants to succeed in gatekeeper courses;(3) to prepare students for research careers via hands-on training in molecular biology techniques and advanced training in systems biology (proteomics, genomics and bioinformatics);(4) to prepare students for a research career by providing workshops in study skills, time management, library research skills, scientific/technical writing, critical literature review, ethics, and competent communication;(5) to provide a selected group of participants with employment as student researchers, addressing research problems particularly focused on diseases, including HIV, diabetes, heart disease, and others, that affect minority populations disproportionately (15 hours per week for 9 academic months per year, 40 hours per week during the summer);(6) to prepare students for graduate school by offering a GRE preparation course and guidance on writing graduate school essays, navigating the application process and developing interview skills so that 100% of participating students apply to and 65% are accepted into a graduate school program;and (7) to provide a selected group of students with the support necessary to succeed in required courses in graduate school and apply for a predoctoral fellowship;90% will complete their dissertation project in a timely fashion. Faculty members from UCDHSC as well as graduate students, fellows and upperclassmen will participate as peer tutors, supplemental instruction facilitators or instructors in the proposed courses/ workshops/ academies. The Chancellor, Provost, Deans and Vice Chancellors have committed funds to support the program and will fund additional slots for highly qualified candidates beyond what has been budgeted in the application, if necessary. Assessment, process and outcome evaluations will be conducted using tools developed by the Associate Dean for Curriculum and Evaluation. We will employ a newly implemented tracking system to measure the impact of the IMSD program on elevating student participant interest in biomedical research and their success in gaining entry into Ph.D.-track programs. Because urban high schools with sizable populations of ethnic and racial minorities have to contend with a myriad of cultural and societal issues affecting basic achievement and academic proficiency, students with underlying talent and capability are too often not afforded the intellectual support available to students from more affluent suburban school districts. We propose to intervene by providing promising students with a program designed to strengthen their fundamental skills, enhance their academic credentials, and elicit their passion for science. Boosting the number of successful biomedical researchers from underrepresented communities will bring much needed diversity to our ranks and potentially encourage greater research interest into the health disparities experienced by these populations.

AIDS Virus; Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; Address; Affect; Alleles; Allelomorphs; Animal Model; Animal Models and Related Studies; Biological Models; CRISP; Cell Growth in Number; Cell Multiplication; Cell Proliferation; Cells; Cellular Proliferation; Chronic; Computer Retrieval of Information on Scientific Projects Database; Conditioned Culture Media; Conditioned Medium; Culture Media, Conditioned; Cytokines, Chemotactic; Development; Disease; Disorder; Employee Strikes; Endothelial Cells; Exposure to; Funding; General Population; General Public; Generalized Growth; Grant; Growth; HIV; HIV-1; HIV-I; HIV1; HTLV-III; Heart failure; Homologous Chemotactic Cytokines; Human; Human Immunodeficiency Viruses; Human T-Cell Leukemia Virus Type III; Human T-Cell Lymphotropic Virus Type III; Human T-Lymphotropic Virus Type III; Human immunodeficiency virus 1; Human, General; Hypertension, Pulmonary; Immune; Immunodeficiency Virus Type 1, Human; In Vitro; Incidence; Infection; Injury; Institution; Intercrines; Investigators; LAV-HTLV-III; Lead; Lesion; Localized; Lung; Lymphadenopathy-Associated Virus; Macaca; Macaque; Mammals, Primates; Man (Taxonomy); Man, Modern; Model System; Modeling; Models, Biologic; Monkeys; NIH; National Institutes of Health; National Institutes of Health (U.S.); Natural History; Pathogenesis; Pb element; Phase; Phenotype; Population; Primates; Property; Property, LOINC Axis 2; Pulmonary Artery; Pulmonary Hypertension; Pulmonary artery structure; Rate; Research; Research Personnel; Research Resources; Researchers; Resources; Respiratory System, Lung; SIS cytokines; SIV; Simian Immunodeficiency Viruses; Source; Strikes; Strikes, Employee; Time; Tissue Growth; United States National Institutes of Health; Vascular remodeling; Viral; Viral Gene Products; Viral Gene Proteins; Viral Proteins; Virus; Virus-HIV; Viruses, General; cardiac failure; chemoattractant cytokine; chemokine; cytokine; disease/disorder; heavy metal Pb; heavy metal lead; human T cell leukemia virus III; human T lymphotropic virus III; macrophage; model organism; mutant; ontogeny; primary pulmonary hypertension; pulmonary; virus protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. According to the world health organization HIV is the number one cause of human morbidity and mortality worldwide. The increased use of highly active antiretroviral therapy in the developed world has resulted in a decrease in morbidity and increase in survival of HIV infected individuals. One consequence of this increased survival is the emergence of new tissue specific manifestations of HIV infection, including pulmonary arterial hypertension. Pulmonary complications of chronic HIV infection are now being recognized with increasing frequency. Currently the pathogenesis of HIV pulmonary arteriopathy is poorly understood, limiting the development of both treatment and prevention strategies. There are numerous limitations in understanding of the relationship between HIV and the development of pulmonary hypertension in humans. These include difficulty in making serial measurements of cardiac and pulmonary function over the natural course of infection, the use of illicit drugs or agents that may be cardiotoxic. Prior studies have shown that rhesus macaques infected with SHIV-nef which is a chimeric viral construct containing the HIV nef gene in a Simian immunodeficiency virus backbone developed complex plexiform [unreadable]like pulmonary arterial lesions similar to those see in HIV associated pulmonary hypertension. In contrast, rhesus macaques infected with a non-chimeric Simian immunodeficiency virus (SIV) did not develop pulmonary arterial lesions similar to those occurring in HIV infected individuals. These findings suggest that the nef gene plays a key role in the development of this life threatening condition. The aim of this study is to try to elicit the cellular mechanisms by which the HIV nef gene induces complex pulmonary artery disease. We hope that by understanding the mechanisms involved in the development of pulmonary vascular disease in SHIV-nef infected macaques that both treatment and prevention strategies for this syndrome can be developed.

DESCRIPTION (provided by applicant): University of Colorado Denver has a successful undergraduate (UG) student diversity research short-term internship program. Better known as GEMS (Graduate Experiences for Multicultural Students), it pairs UG students from diverse and disadvantaged backgrounds with lab mentors. Over the past 10 years, 101 UG students participated: 27 are still UG (85% will apply to professional schools);74 have graduated, 49 are in some health professional school or have a health-professional degree for a success rate of 66%. The Clinical Translational Science Award (CTSA) launched the Colorado Clinical Translational Science Institute (CCTSI) to build infrastructure for educational resources/enhancement of translational research. The CCTSI will partner with the GEMS program. In this new R25 we request funds to continue (and replace) the 10 year-old NHLBI-funded T35 GEMS and provide experiences to 10 UG and 2 health professional students. We propose several aims: 1.expose students to research careers by immersion in laboratories with basic or physician scientists;2. expose students to quantitative and analytical skills;3. improve student comprehension of science by participation in brown bags covering several topics;4. expose students to science topics through research seminars;5. increase awareness of clinical/translational research by shadowing physician scientists at the clinic;6. create a website in partnership with the CCTSI to track students. With this award, we will continue to stimulate students from diverse backgrounds to pursue health-related careers. PUBLIC HEALTH RELEVANCE: This internship program at the University of Colorado Denver, Anschutz Medical Campus, seeks to expose undergraduate and health professional students from diverse and disadvantaged backgrounds to summer laboratory or clinical translational research experiences in order to increase their representation in research related to cardiovascular, pulmonary or hematologic areas. The goal is to create a workforce of health-related researchers who are as diverse as the community they serve. By exposing these students early in their careers, we hope to capture their interest and excite them about the possibilities of biomedical research. (End of Abstract)

DESCRIPTION (provided by applicant): As HIV-infected individuals continue to age, non-infectious complications increase in frequency. Despite the first descriptions of patient cases in 1987, little is known about the pathogenesis of pulmonary hypertension associated with HIV (PAH-HIV). Chronic exposure to viral products such as HIV-1 Nef and HIV-induced immune deregulation in the lung may contribute to pulmonary vascular disease, particularly through their impact on pulmonary endothelial cells (EC). Our group was the first to associate the HIV-1 nef protein with the pathogenesis of vascular remodeling PAH-HIV. We longitudinally followed 34 individuals with PAH-HIV and collected sequential clinical and echocardiographic; we created and curated a repository of plasma, bronchial lavage fluid and cells. We sequenced the nef gene from blood and lung samples from patients with PAH, elevated pulmonary artery systolic pressures and from non- PAH HIV infected. We found amino acid substitutions in the Nef protein statistically associated with the PAH phenotype; these substitutions clustered around Nef functional domains that potentially interfere with Nef adaptor functions. Further studies, which are preliminarily reported in this application, suggest that particular amino acid signatures predominate in the lungs compared with the periphery. The lung may be a protected environment that allows the virus to evade immune responses; furthermore, particular nef alleles will enhance T cell responses and result in pulmonary vascular endothelial cell proliferation/apoptosis, disrupt signal transduction pathways and result in vascular remodeling. We hypothesize that allelic variants of nef will have an impact on T cell and pulmonary endothelial cell function. We propose to 1) infect T cells with molecularly cloned virions containing primary nef alleles and measure T cell receptor density; 2) human pulmonary artery endothelial cells will be co-cultured with infected T cells or transfected with nef molecular clone and endothelial cell gene expression, apoptosis and proliferation measured and 3) determine whether the lung is a protected compartment for evolution of nef sequences. These studies will examine the functional properties of nef alleles containing these amino acid substitutions from PAH-HIV individuals. The studies proposed in this application will use existing biospecimens and cloned nef alleles to examine the mechanisms whereby HIV-nef influences pulmonary vascular remodeling in the pathogenesis of PAH-HIV. Our research tem, with a combination of basic and clinician scientists is well poised to address how the nef viral protein and immune dysregulation are contributing factors to this lung complication of HIV.

DESCRIPTION (provided by applicant): As the face of the United States becomes more diverse, it is imperative that institutions of higher education increase the number of PhDs awarded to students who are members of underrepresented minority groups. The University of Colorado at Denver (UC-Denver) is committed to this goal and is in an excellent position to enhance and intensify its training of undergraduate students to foster diversity not only in the PhD workforce in higher education, but also in research beyond the University setting. This MARC U*STAR program application supports these efforts by providing intensive interdisciplinary curricular and research training for 25 URM students during the last two years of their undergraduate education that will enable them to enter PhD programs in basic and applied behavioral science and pursue research careers in these fields. The regional and local demographics of UC Denver along with the rapid expansion of research in the behavioral sciences and an already thriving undergraduate program provide a strong foundation for building the proposed MARC U*STAR Scholars program in behavioral sciences. The proposed program features a new series of courses in the methodology and ethics of behavioral and health science research as well as on and off- campus research experiences that will provide the strong academic and experiential foundation necessary for students to gain entrance into and succeed in PhD programs. Importantly, these activities leverage other minority training programs at UC Denver to expand an institutional culture that supports the success of minority students and their entrance into graduate education in the sciences. The MARC program, which builds on existing interdisciplinary degree programs in Psychology and Public Health, enjoys strong institutional support and holds the potential to be transformational for the climate of diversity and inclusion at UC Denver.

DESCRIPTION (provided by applicant): This is a competing renewal application for a short-term research education program originally funded as a T35 in 2000 and as an R25 in 2010. Our major objective is to continue providing annual short-term research education experiences for highly motivated minority students in order to expose them to biomedical research in the area of pulmonary and cardiovascular disease. Locally known as GEMS (Graduate Experiences for Multicultural Students). Over the past 13 years, >170 UG students participated; >115 supported by the T35/R25 and the rest supported by other programs. Collectively, these students have published 81 manuscripts; >60% have earned terminal degrees or are enrolled in professional school. More than 70% of student participants were under-represented ethnic minorities. The program builds upon our established infrastructure and uses the significant strengths of one of the top pulmonary medicine programs in the country. We continue the tradition of addressing the pipeline by requesting 10 undergraduate and 4 health professional student slots. We will also emphasize the concept of team science that is so critical to success in the biomedical research field. Here, in addition to the usual didactic and hands-on research activities, we propose a new model of training that deviates from the traditional mentor/mentee relationship; a paradigm shift where we introduce the concept of academic coaches. A coach is not intended to supplant the mentor, but rather complement this relationship, since coaches do not interact with the student on a day-to-day basis. Coaches are experienced faculty members who guide the students through a successful career path and stay in contact beyond the summer internship. Furthermore, to ensure student success, we propose to use social science approaches and provide the students with a toolkit that will create an environment, a community of practice, where they feel safe to talk about personal, academic and professional issues and to bond through shared norms and values. We will use traditional approaches to student selection but will incorporate a set of targeted questions in the application that will aidin selection of students highly motivated to pursue biomedical research. We believe that these approaches will continue the GEMS tradition of excellence in training under- represented students while at the same time enhancing student's academic success beyond the summer GEMS internship. (End of Abstract)

PROJECT SUMMARY As the United States becomes more diverse, it is imperative that institutions of higher education accelerate their efforts to ensure equity in the number of doctoral degrees awarded to students who are members of underrepresented (UR) groups. The University of Colorado Denver (CU Denver) is committed to this goal, and uniquely situated culturally and geographically to meet this need. The CU Denver campus is consolidated with the Anschutz Medical Campus (Anschutz) and, together, is classified as a ?Doctoral University: Higher Research Activity?, with more than $400 million in annual sponsored research funding. As such, CU Denver is the only urban public research university in the State of Colorado, and it has the most diverse student body of any University of Colorado campus. In 2016, at least 34% of entering freshmen identified as underrepresented minority (URM) students and at least 57% as students of color. CU Denver was awarded its first MARC U-STAR award on May 22, 2013 (NIGMS T34 GM096958; 2013-2018), and is one of only two in the state. This competing renewal expands the successful initial development of the CU Denver MARC U-STAR program in several important ways. First, it broadens the focus of the first award (supporting students in the biomedical- oriented behavioral sciences) to the biomedical sciences more broadly by providing 24-month support to 40 additional rising junior undergraduates (eight new scholars per year) majoring in Integrative Biology, Chemistry, Psychology, and Public Health. Second, it expands and further institutionalizes a series of successful undergraduate Behavioral and Biomedical Research training courses, developed for the MARC program and taken by all MARC scholars, to accompany and facilitate their authentic research experiences. Third, it leverages and supports an institutionally-funded pre-MARC ?U-RISE? program, developed by the principal investigators and program staff, to identify and support freshman, sophomore, and transfer students with authentic research experiences that engage early interests in research careers. The CU Denver MARC program complements and extends other training programs at CU Denver and Anschutz, expanding an institutional culture that supports the success of UR students and their entrance into doctoral programs in the biomedical sciences. The new efforts described here, focused on expanding our initial successful work with recruitment and training of MARC scholars from the Psychology Department, and building the base of students interested in these opportunities, are complemented by a rigorous, external program evaluation that continuously assesses the effectiveness and relevance of our program activities, and includes intentional outcomes in program optimization and scholarly dissemination. The MARC program enjoys strong institutional support and has already helped transform the climate of diversity and inclusion at CU Denver. These efforts, together with an outstanding research environment and culture of support for diversity and inclusion, will help the CU Denver MARC U-STAR program continue to meet the overarching MARC program goal that 90% of supported students graduate with a STEM degree and at least 60% will matriculate into Ph.D. (or combined M.D./Ph.D.) programs in the biomedical sciences with a completion rate of at least 80%.

PROJECT ABSTRACT This is a new application for a ?PRIDE Academy: Impact of Ancestry and Gender on omics of lung diseases? whose objective is to introduce scholars from under-represented backgrounds to omics of lung diseases with a focus on how datasets should be interpreted and applied when working with under-represented populations. This academy will be housed at the Pulmonary and Biomedical Informatics and Personalized Medicine (BiPM) Divisions at Anschutz Medical Campus in Colorado. The Pulmonary Division has a distinguished historical record of training leaders in pulmonary medicine and the relatively new BiPM Division has been instrumental in elucidating to what extent genetics can explain health disparities in complex diseases, particularly asthma. We propose a PRIDE summer academy that will include didactic and hands-on workshops in genomics and proteomics of lung diseases such as asthma, emphysema, pulmonary fibrosis, and pulmonary hypertension. The PRIDE scholars will be assigned a mentoring team closely aligned with the mentees? research interests. Here, in addition to the usual didactic and hands-on activities, we propose an additional level of training that uses the concept of academic ?coaches?. A coach is not intended to supplant the mentor, but rather complement this relationship, since coaches do not interact with the scholars one-on-one but rather as a group. The coach will guide team members through the process of successfully navigating the academic world using well-tested social science approaches. The overall objectives are to select talented underrepresented junior faculty scholars with a demonstrated interest in pulmonary diseases; introduce the scholars to a toolkit including a mentoring team, instruction on ?omics? of cardio-pulmonary diseases and a suite of social science theories for academic persistence; pair coaches with scholars to ensure that all the milestones are achieved; provide scholars with grant-writing workshops and mock study sections that will position them to compete for intramural and extramural grants; implement an evaluation plan that measures the degree to which the program is achieving its objectives. Scholars will travel to Denver for the PRIDE summer academies for 2 consecutive years and to Aspen for a face-to-face mid-year meeting at the Annual Tom Petty Aspen Lung Conference. By using the combination of coaching and social science practices such as cultural capital and communities of practice that allow team members to feel connected with each other, and pairing the scholars with distinguished pulmonary and BiPM faculty mentors, we will ensure their academic persistence and ultimate success.

PROJECT SUMMARY As the United States becomes more diverse, it is imperative that institutions of higher education accelerate their efforts to ensure equity in the number of doctoral degrees awarded to students who are members of underrepresented (UR) groups. The University of Colorado Denver (CU Denver) is committed to this goal, and uniquely situated culturally and geographically to meet this need. The CU Denver campus is consolidated with the Anschutz Medical Campus (Anschutz) and, together, is classified as a ?Doctoral University: Higher Research Activity?, with more than $400 million in annual sponsored research funding. As such, CU Denver is the only urban public research university in the State of Colorado, and it has the most diverse student body of any University of Colorado campus. In 2016, at least 34% of entering freshmen identified as underrepresented minority (URM) students and at least 57% as students of color. CU Denver was awarded its first MARC U-STAR award on May 22, 2013 (NIGMS T34 GM096958; 2013-2018), and is one of only two in the state. CU Denver was awarded a competing renewal in 2016. The renewal expanded the successful initial development of the CU Denver MARC U-STAR program to support 20 additional rising junior undergraduates (four new scholars per year) majoring in Integrative Biology, Chemistry, Psychology, and Public Health. The CU Denver MARC program complements and extends other training programs at CU Denver and Anschutz, expanding an institutional culture that supports the success of UR students and their entrance into doctoral programs in the biomedical sciences. One unique aspect of the CU Denver MARC U-STAR program is a partnership with the CU Denver Clinical Health Psychology (CHP) program. Through this partnership, we have retained an advanced CHP doctoral student to serve as a wellness point-person for our scholars. This doctoral level student addresses threats to wellness and resiliency on multiple levels using preventative, in- vivo, and follow-up communication aimed at detecting scholar distress and building adaptive copings skills to mitigate distress for our scholars. This supplement aims to adapt this evidence-based program to be more culturally-responsive through a formative needs assessment (barriers impacting scholars, supports needed, self-advocacy skill development) and integration of new learning with the wellness and resiliency program. Culturally-responsive wellness and resiliency training can provide minoritized scholars with knowledge and skills necessary to successfully navigate academic adversity and discrimination with self-advocacy and resilience. These efforts, together with outstanding academic and research training and a culture of support for diversity and inclusion, will help the CU Denver MARC U-STAR program continue to meet the overarching MARC program goal that at least 90% of supported students graduate with a STEM degree and at least 60% matriculate into Ph.D. (or combined M.D./Ph.D.) programs in the biomedical sciences with a doctoral completion rate of at least 80%.