Rutgers Biomedical And Health Sciences

NIH Research Projects · FY 2024 · 2020-09

T cells play a critical role eliminating pathogens and the generation of memory T cells is an important component in protection from secondary infection. Memory T cells can be divided into two groups based on their location, those that are capable of circulating throughout the body and those that are lodged in tissues, poised to respond rapidly to secondary infection. Tissue-resident memory T cells (Trm) cells remain in the tissue and are not replenished by circulating cells after infection is resolved. Circulating T cells are often not sufficient to protect from secondary infection; therefore, it is of significant interest to determine how Trm cells are generated and maintain their function over time. Only a small number of microbes need to breach the mucosal surface to initiate disease. The ability of adaptive immune cells to locate pathogens in large, complex tissues and eliminate them before they disseminate to deeper tissues is a necessary component of protective immunity. We have used infection with the intestinal pathogen Yersinia pseudotuberculosis to examine pathogen-specific CD8+ Trm during infection, and using this model we have uncovered significant phenotypic heterogeneity in intestinal Trm cells, with expression of the integrin CD103 defining these populations. Proximity of T cells to areas of infection within the intestinal tissue regulates Trm differentiation, with inflammation and activation of the transcription factor STAT4 leading to increased numbers of CD103 Trm cells. This proposal will identify the underlying mechanisms that regulate the differentiation and maintenance of the CD103 Trm subset. We have already shown a critical role for CD103 Trm cells in controlling pathogen replication during primary infection, and we have developed new tools to analyze the division of labor between Trm subsets during secondary infection. These findings will address a fundamental gap in our knowledge regarding the function of Trm cells in controlling intestinal colonization during secondary infection. Additionally, it is currently unclear whether either Trm subset alone is sufficient to confer protection, and we will determine if the full complement of Trm cells is necessary for robust immunity. This work will identify strategies to maximize the number and persistence of Trm cells, an important component of any successful vaccination strategy to target mucosal pathogens.

NIH Research Projects · FY 2024 · 2020-09

Project Summary/Abstract Candidate: My career goal is to develop an independent research program that examines how hierarchical transformations of auditory information downstream of core auditory cortex supports sound-driven decisions. My previous training experiences has provided me with technical and intellectual skills on auditory neurophysiology, behavioral neuroscience, and hearing loss-induced deficits in auditory processing and perception. I propose to expand my skill set with additional training by conducting wireless cortical recordings while gerbils simultaneously perform a complex auditory task, pharmacological and chemogenetic attenuation of neural activity in awake- behaving animals, and statistical modeling of psychophysical and neural data. During the K99 phase, I will prepare my transition to independence by developing the necessary skills for producing a successful chalk talk, plan job interview strategies, and gain effective lab management applications. By the end of the R00 phase, I plan to have a strong publication record and ample preliminary data that will lead to a successful R01 application. Environment: The K99 phase of this will be award will take place in the Center for Neural Science at New York University; an ideal location for the proposed training phase of this award. My primary mentor, Dr. Dan Sanes, possesses 30+ years of experience in developmental plasticity and auditory neuroscience. His laboratory utilizes innovative physiological, pharmacological, and anatomical techniques that include in vitro slice physiology, wireless extracellular awake-behaving recordings, pharmacological manipulation of neural activity, optogenetics, and anatomical tract tracing with viral vectors. Additional mentoring will be provided by Dr. Xiaoqin Wang (John Hopkins University), who has 25+ years of experience as an expert in the neural coding of acoustic information along the auditory neuroaxis and neurophysiological recordings in awake preparations, and Dr. Roozbeh Kiani (New York University), who is an expert in the neural mechanisms of perceptual decision-making. Research: The neural representations of acoustic signals are transformed at each locus of an ascending auditory pathway. An example of a principal characteristic of this hierarchical processing is the increase of integration time (i.e., the time required to fully encode a sensory cue) across higher auditory cortices. Currently, it is less certain how auditory information is transformed downstream from core auditory cortex (ACx) where it becomes integrated with other sensory inputs, and supports sound-driven decisions. Perturbations of neural activity in parietal cortex (PC), a region downstream of ACx, will be made in animals performing an auditory temporal integration task to determine whether PC is necessary for the observed behavioral temporal integration times (K99). Wireless recordings will be made in ACx and PC from awake-behaving animals performing an auditory temporal integration task to reveal the neural representations that support perceptual integration times (K99). Similar approaches will be used to determine whether the neural representation of PC correlates and supports task performance during a combined multisensory condition (R00).

NIH Research Projects · FY 2026 · 2020-09

Project Summary. New Jersey has significant cancer disparities across certain population groups, which also remain underrepresented in the cancer research and healthcare workforce. Increasing workforce diversity is one of the key strategies for reducing cancer inequities. In alignment with the National Cancer Institute’s (NCI) mission to train and educate a diverse workforce to reduce cancer health disparities, the goal of the Rutgers Youth Enjoy Science (RUYES) program is to increase the diversity of the biomedical, specifically cancer research workforce. The ongoing RUYES program at the Rutgers Cancer Institute is funded by the NCI grant R25 CA247785 awarded in September 2020. In the current funding period, we have successfully engaged 58 participants, including high school science teachers, high school and undergraduate students, in mentored cancer research, professional development, and community outreach since the summer of 2021. The outreach initiatives undertaken by RUYES participants have positively impacted thousands of individuals within their families and communities, ultimately bolstering the importance of science education and careers. This renewal application introduces new components and learning opportunities to further enhance our highly successful RUYES program. These additions are informed by valuable lessons learned and stakeholder feedback. In the next funding period, we propose a holistic program that focuses on student success, includes new research experiences in Population Sciences and Cancer Health Equity, a virtual Community Scientist Training Program, and partnerships with Minority-Serving Institutions (MSI) and Historically Black Colleges and Universities (HBCUs). We expand our approach to 1) Provide robust support to middle school and high school science teachers and undergraduate faculty in MSI and HBCUs, offering mentored cancer research experience and comprehensive help with curriculum development; 2) Actively engage high school and undergraduate students from diverse backgrounds in mentored hands-on cancer research, as well as provide extensive co- curricular and professional career development activities; 3) Create dynamic and impactful cancer-related outreach activities that actively connect program participants with schools and families in their communities. In our renewal proposal, we will leverage our experience and track record of highly successful pathway programs with excellent outcomes, a multi-disciplinary research environment, and robust educational and outreach resources of our Cancer Institute, the state’s only NCI-designated Comprehensive Cancer Center. Our strategically forged synergistic partnerships with similar initiatives across and beyond Rutgers will ensure that RUYES continues to expand on its achievements in preparing the next generation of a diverse cancer research and healthcare workforce.

NIH Research Projects · FY 2025 · 2020-08

PROJECT SUMMARY/ABSTRACT The goals of this project are to further our understanding of the long-term clinical epidemiology of health outcomes among drug users who are at high risk for HIV. This is important because it will guide clinical practice and health systems planning in the face of the continuing HIV/AIDS epidemic and the evolving epidemic of drug abuse, including opioids. Many HIV-infected persons are living longer than they had in the past, due to the advent and availability of highly active antiretroviral therapy. We have a unique prospective cohort of drug users established in the 1980s, backed by a linked biospecimen repository and permission for indefinite continuing follow-up, established by the NCI and NIDA for the long- term study of drug users. The cohort numbers nearly 11,000 drug users, who are diverse in gender, race, ethnicity and geography, and include large numbers of both HIV-positive and HIV-negative persons. Hepatitis C virus infection is extremely common in these persons. WE SEEK TO UNDERSTAND THE HEALTH OUTCOMES IN THIS COHORT THROUGH TWO AIMS. AIM 1. To determine the epidemiologic features of mortality outcomes associated with HIV among drug users over a 35-year period. This will be accomplished by matching our cohort to vital statistics registry data. AIM 2. To determine the cancer epidemiology, by cancer site, among drug users over a 35-year period, including incidence, stage at diagnosis, treatment regimens, mortality, and HIV-associated risk. This will be accomplished by matching our cohort to cancer registry data. Our project analyses will integrate these two types of data, with stepwise repetition of matching over several years. Mortality data will enable us to more accurately conduct survival analyses on an array of disease outcomes, with a special focus on cancer to identify putative risk factors. This has the potential to assist with the identification of interventions that would reduce disease burden and prolong survival. We will characterize disease outcomes in HIV-positive compared to HIV-negative drug users, accounting for baseline demographic, behavioral, and medical factors. Our cohort uniquely affords the opportunity to study health outcomes over a prolonged time period spanning several decades in both sexes and in underrepresented minority populations. The size of our study and the extent and richness of baseline data will make it a powerful tool in understanding a wide array of disease outcomes. Given the continuing rapid evolution of biotechnology, future research might employ evolving biotechnology to use our biospecimen repository to explore specific disease outcomes we will be identifying. Our cohort has joined the NIDA Genetics Consortium. We will explore joining other consortia which focus on other outcomes, which could amplify the power of those consortia.

NIH Research Projects · FY 2024 · 2020-07

Project Summary This proposal is a summer research and education program for health professional students in areas related to the research mission of the NHLBI. Mentored by faculty drawn from selected health professional schools of Rutgers, the program has three primary aims: (1) to increase student knowledge and appreciation of biomedical research by providing a closely-mentored, hands-on research experience; (2) to increase student knowledge and interest in pursuing careers in research in mission-related areas of the NHLBI through career development and educational activities; (3) to provide continued advice, support and guidance to program alumni to facilitate post-program career planning and implementation. Ten health professional students from Rutgers New Jersey Medical School (NJMS), Robert Wood Johnson Medical School (RWJMS), School of Dental Medicine (SDM) and School of Health Professions (SPH) will be admitted to our program each year beginning in 2020. Special consideration will be given to students from economically and/or socially disadvantaged backgrounds, including underrepresented minority applicants and those with limited research experience. Our selection of participants is enhanced by the proven track record in diversity recruitment as demonstrated by the ranking of Rutgers NJMS in the 80th percentile in underrepresented minority students and 85th percentile for minority faculty, according to the 2014 Association of American Medical Colleges’ Mission Management Tool. The research component of the proposed program is aligned with the training program being implemented in a parallel pending Clinical and Translational Science Award program to Rutgers. The R25 program will provide hands-on experimentation in labs with active, nationally recognized and funded research programs at Rutgers in mission areas of the NHLBI, specifically studies related to cardiovascular, pulmonary and hematological diseases. These students will be exposed to high-level research using cutting- edge, experimental strategies and contemporary methodology. The infrastructure and organization of the proposed R25 summer program will take advantage of the leadership, teaching, research knowledge and skills of faculty who have extensive experience in training health professional students. The program is designed to integrate student research experiences and mentoring with weekly meetings comprising training in science communication, critical reading of manuscripts and Responsible Conduct of Research. Each year, students will present the results of their summer research projects in a symposium and submit a paper on their project in the format of a manuscript. Contact by program directors with alumni via social network platforms and in- person reunion events will provide continued career-related advice, mentoring and networking of prospective, current and past participants who can share their experiences and advise each other.

NIH Research Projects · FY 2024 · 2020-07

PROJECT SUMMARY: Over the past two decades, high rates of opioid prescribing have led to widespread opioid use and epidemic levels of adverse opioid-related outcomes. Early interventions focused on reducing opioid prescribing through the development of practice guidelines and implementation of programs and policies to increase prescription opioid management and monitoring. While overall opioid prescribing has declined in recent years and may have contributed to reductions in prescription opioid-involved overdose death rates in 2012 and 2013, the rate has since risen to a new high, comprising 40% of all opioid-related deaths in 2016. Furthermore, prescribing remains elevated in disadvantaged groups and areas, highlighting a need to better understand how individual characteristics intersect with health system and population-level factors in the pathways from opioid use to fatal and non-fatal overdose. Using a multi-level approach, the proposed project examines trajectories from opioid treatment for chronic non-cancer pain to opioid overdose and fatality, accounting for: 1) individual sociodemographic and clinical characteristics (e.g. sex, age, race/ethnicity, comorbidities), 2) opioid treatment patterns (e.g. dosage, duration, formulation), 3) county-level health system (e.g. facilities, services) and population characteristics (e.g. percent poverty, minority, unemployed, disabled), and 4) state-level policies (e.g. Medicaid policies, Prescription Drug Monitoring Programs). Opioid treatment patterns will be assessed using national Medicaid Analytic Extract (MAX) data, adjusting for individual characteristics. Risk of overdose based on individual characteristics and treatment patterns will be assessed using MAX data for medically treated non-fatal opioid overdose and National Death Index (NDI) data for fatal opioid overdose. Overdose outcomes will be further assessed using county-level Area Health Resource File (AHRF) data and state-level policy measures. Contextualizing individual outcomes within wider health care systems and social and geographic settings supports NIDA’s strategic plan to increase the public health impact of research. Additional substantive and methodological training will contribute to the applicant’s long-term goal to develop an independent academic research career and generate and disseminate robust evidence to inform effective interventions to reduce opioid-related morbidity and mortality. The training aims include advanced study and application of: 1) innovative methods to characterize clinical care patterns over time, 2) social and spatial epidemiology theory and methods to study multilevel influences on health, 3) policy drivers of health and health care, and 4) professional development for conducting ethical, independent research. The research and training are supported by an interdisciplinary mentorship team of Columbia University researchers with expertise in substance use and policy research, clinical and spatial epidemiology, and statistics with additional support from external collaborators in health services and policy research methods. This project and career development plan will prepare the applicant to secure R01 funding to extend this line of research.

NIH Research Projects · FY 2024 · 2020-06

ABSTRACT In response to RFA-AI-19-030, Feasibility of Novel Diagnostics for TB in Endemic Countries (FEND-TB) the leadership team has brought together a consortium of experienced investigators and clinical sites and developed a research plan to address critical unmet TB diagnostic needs. This program benefits from experience gained during the successful 7-year tenure of the NIH DMID-funded TB-Clinical Diagnostics Research Consortium (TB-CDRC), with overlap in leadership, investigators and sites. This program has been adapted in several ways to further enhance capacity to meet the current challenges in the field -- the successful collaboration with the Foundation for Innovative New Diagnostics (FIND) has been strengthened to now a full partnership that will facilitate access to cutting edge technologies and alignment of FEND-TB work with global stakeholder priorities; clinical study sites in India and Peru have been added to accelerate recruitment and augment capacity to enroll patients with co-morbidities and drug- resistance; inclusion of a mature analytic laboratory and revised technology evaluation strategy that together allow for rational, nimble, step-wise evaluation of early-stage diagnostics; and inclusion of mathematical modeling capacity to inform optimal diagnostic strategies in TB endemic settings. This proposal will test two main hypotheses: A. Novel early stage TB diagnostics, that target bacterial and/or host targets and will be ready for evaluation in the next five years, will have performance characteristics suitable for point of care (POC)/near-care use for TB detection, triage, or drug susceptibility testing. B. Rapid non-sputum diagnostics will provide ancillary support as components of algorithms for the diagnosis of childhood TB as well as paucibacillary pulmonary TB and extrapulmonary TB in adults. Specific Aims are: 1. To evaluate the diagnostic accuracy of early stage diagnostic tests for tuberculosis. 2. To identify new early stage diagnostics for evaluation, and to develop and implement for each a stepwise evaluation plan. 3. To use economic analysis and transmission modelling to design optimal diagnostic algorithms.

NIH Research Projects · FY 2026 · 2020-04

Project Summary/Abstract Young adults with cancer experience more emotional distress and greater impairments in health-related quality of life (HRQOL) than older patients. Cancer diagnosis during young adulthood interferes with participation in normal developmental tasks such as obtaining higher education, starting a career, establishing financial independence, and developing romantic partnerships. This disruption in normal activities coupled with the unfamiliar and challenging demands of cancer treatment results in emotional distress and reduced HRQOL. There is an urgent and critical need to develop, test, and implement evidence-based interventions to support these young adults as they navigate perhaps the most challenging and debilitating period of their lives. Current psychosocial care does not adequately address the unique concerns of young adults. An optimal solution would give young adults the skills to deal with diverse and numerous stressors, address underdeveloped problem-solving ability characteristic of this age group, and be relatively simple to learn and use during the highly stressful time following a diagnosis of cancer. To address these clinical care gaps, the parent award is evaluating the efficacy of a problem-solving skills training intervention developed specifically for young adults and grounded in the core tenets of problem-solving therapy. “Bright IDEAS-Young Adults” (Bright IDEAS-YA) draws upon and notably extends prior research demonstrating the efficacy of a cognitive-behavioral problem-solving skills training program for reducing emotional distress in caregivers of pediatric patients. Bright IDEAS-YA is a 6-session, one- on-one intervention that teaches patients a systematic approach to overcome personal challenges across any life domain. It aims to enhance patients’ problem-solving ability in the face of significant stressors such as cancer. In preliminary work, young adults with cancer found Bright IDEAS-YA acceptable, relevant, and useful. Patients who received Bright IDEAS-YA showed improvements in problem-solving ability and reductions in symptoms of depression and anxiety. In the parent award, we are currently conducting a multi-site randomized controlled trial of Bright IDEAS-YA compared with enhanced usual psychosocial care with 344 YA patients (ages 18-39 years) undergoing cancer treatment. We will evaluate efficacy and examine mediators and moderators of intervention effects using assessments at baseline, post-intervention (3 months), and follow-up (6 and 12 months). In the proposed extension period, we will evaluate the sustainability of intervention effects by extending the follow-up to 2-years post-enrollment; identify barriers and facilitators of integrating Bright IDEAS-YA into clinical practice; and address barriers to access by culturally and linguistically translating for Hispanic young adults.

NIH Research Projects · FY 2025 · 2020-04

ABSTRACT Halting tuberculosis (TB) transmission will be most efficiently achieved by focusing control efforts on sites of high transmission and by developing new drugs and vaccines that target bacterial transmission. We hypothesize that household TB transmission is dramatically under-estimated because mixed infections are common and secondary cases may be infected by minor populations having a transmission advantage; ergo that mismatch between the Mycobacterium tuberculosis (Mtb) strain isolated from the index case and secondary case does not preclude household transmission. We also hypothesize that the host immune and pathological response to Mtb isolates and the ability of different Mtb isolates to survive the stress of aerosolization strongly impacts transmission potential. Here, we propose to study characteristics of TB transmission within households, the bacterial isolate and its interaction with the host innate and adaptive immunity that aid or are permissive of transmissibility. To address these hypotheses, we will conduct retrospective and prospective cohort studies in Brazil and in Uganda leveraging over a decade of household contact studies in Brazil (2004-date) funded by the International Collaboration for Infectious Diseases Research (ICIDR) and the Tuberculosis Research Unit (TBRU), and over 20 years of collaborative research among our study team. Our Specific Aims are: 1) To determine the extent to which household transmission is responsible for co-prevalent and incident TB cases that initially appear to be genetically distinct from index cases within the same household. 2) To investigate the in vitro immune phenotype of transmitted versus not-transmitted Mtb isolates in Brazilian and Ugandan households stratifying for HIV status. 3) To characterize the innate and adaptive immune response in persons with known household exposure to high and low transmission isolates and the impact of HIV infection. 4) To identify the component of Mtb genes that aid in bacterial survival in aerosols. 5) To investigate the bacterial factors underlying divergence in the host response to high and low transmission isolates. Together, these studies will provide new paradigms for transmission and novel targets for intervention.

NIH Research Projects · FY 2025 · 2020-04

PROJECT SUMMARY Cell fate is determined by a number of factors including genetics, cellular signals and availability of nutrients. How cells process these numerous inputs to produce a specific output, such as cell differentiation or proliferation remains poorly understood. Our project will address how cells process these inputs following the principle of “supply and demand.” During early T cell development, the synthesis of a diverse repertoire of T cell receptors defines a robust immune system that will allow recognition of various pathogens, while tolerant to self-peptides. The synthesis of a diverse repertoire requires abundant nucleotides and hexosamines for proper synthesis and folding. Our studies will elucidate how the de novo hexosamine biosynthesis pathway reprogram metabolism via regulation of mTOR signaling and pyrimidine metabolism to generate a diverse TCR repertoire. As we gain better understanding of how these pathways are reprogrammed to balance nutrient supply with cellular demand, we hope to develop better strategies for dietary manipulation that can potentially enhance the functions of the immune system.

NIH Research Projects · FY 2026 · 2020-02

The USFDA approval of the α particle emitting radiopharmaceutical (RP) radium 223 dichloride (Xofigo®) and the β-particle emitting lutetium 177 dotatate (LUTATHERA®), and their successful implementation in the clinic, has contributed to reinvigorated interest in radiopharmaceutical therapy (RPT) of cancer. RPT entails the delivery of radioactive drugs to the primary tumor, metastases, and disseminated tumor cells (DTC). Different classes of radionuclides have been advocated for therapy including α , β , and Auger emitters. The different ranges of these radiations in tissue, and their differences in relative biological effectiveness (RBE), contribute to the complexity of predicting therapeutic efficacy. ‐However, like external beam radiation therapy, the future of RPT will depend in part on our capacity to plan treatments that maximize therapeutic effect while minimizing adverse effects in normal tissues. Key to the long term success of RPT is to overcome limitations of the intrinsic nonuniform uptake of the radiopharmaceutical by cancer cells that can impact our capacity to sterilize tumors, metastases, and DTC. While primary tumors can often be addressed with external beams of radiation, micrometastases and DTC cannot. While there are commercial tools to assist with calculating absorbed dose to macroscopic disease based on external imaging and using it to predict response, there is a dearth of tools that can be used to optimize and plan RPT of microscopic disease. Only MIRDcell V2, developed in the Howell lab in collaboration with the MIRD Committee in 2014, is widely available. MIRDcell V2 has strengths and weaknesses. This project seeks to overcome many of the weaknesses by creating MIRDcell V3 with new capabilities to facilitate RPT design and treatment planning of micrometastases and DTC. In addition, MIRDcell V3 will serve as an indispensable educational tool for dosimetry and radiobiology of radiopharmaceuticals. Students will be able to operate MIRDcell V3 and learn about how the selection of different radionuclides and other parameters are expected to affect cell killing. The influence of particle range, RBE, activity distribution and other parameters can be explored. In view of the new research that was spurred by its predecessor, MIRDcell V2, this educational element is perhaps one of the most important aspects of MIRDcell V3.

NIH Research Projects · FY 2025 · 2020-02

Abstract Lung cancer is the leading cause of cancer mortality, with non-small cell lung cancer (NSCLC) accounting for more than 85% of these cases. KRAS, the most common oncogenic driver in NSCLC, confers a poor prognosis with limited treatment options. LKB1 is the third most frequently mutated gene in NSCLC. The mutations in both KRAS and LKB1 account for about 30% of NSCLC, with increased aggressiveness, a high frequency of metastases and resistance to therapeutics. Autophagy degrades proteins and organelles and recycles them to provide metabolic substrates, a function that is critical when extracellular nutrients are limited. Although the role of autophagy in cancer has been intensively studied, the precise role of autophagy in cancer, especially in vivo, remains elusive and controversial. Moreover, targeting autophagy to treat cancer generally has not contributed significantly to the advancement of clinical trials. Therefore, identifying genetic vulnerability that renders strong sensitivity to autophagy inhibition is urgently needed to improve autophagy targeted- therapies. LKB1 regulates energy homeostasis by activating AMP-activated protein kinase (AMPK), which inhibits catabolic processes and upregulates anabolic processes, in response to energy crisis. Based on earlier studies, we began to test the hypothesis that loss of LKB1 promotes cancer cell proliferation but also restricts adaptation to metabolic stress, a property that may be further compromised by loss of autophagy. Using genetically engineered mouse models (GEMMs) of NSCLC, we found that autophagy inhibition was synthetically lethal in KrasG12D/+;Lkb1-/- (KL) mediated tumorigenesis; in contrast to KL lung tumors with intact autophagy, loss of an essential autophagy gene, Atg7, dramatically impaired both tumor initiation and tumor growth. This is in sharp contrast to wild-type LKB1 (KrasG12D/+;p53-/-) tumors that are much less sensitive to essential autophagy gene ablation. Our in vitro study further revealed that autophagy modulates lipid metabolism essential for KL cancer cells to survive nutrient starvation. These observations indicate that LKB1 mutations predispose KRAS- driven NSCLC to autophagy inhibition and that LKB1 mutations could be explored as a predictive biomarker for precision lung cancer therapy. Based on our recent findings, we form our central hypothesis: autophagy compensates for LKB1 loss by maintaining the metabolism of Lkb1-deficient Kras-driven lung tumors and promoting their metastasis. We will test this with following specific aims: Aim 1. Elucidate the mechanism by which autophagy regulates lipid metabolism and KL tumorigenesis in vivo. Aim 2. Determine how autophagy promotes KL tumor metastasis. Aim 3. Identify metabolic bypasses that potentially create resistance to autophagy inhibition in KL NSCLC. Successful completion of this proposal will: (1) yield new insights into the role of autophagy in modulating cellular metabolism in support of KL lung tumorigenesis and metastasis; (2) validate the novel concept that autophagy inhibition is a selective and powerful therapeutic strategy against primary and metastatic KL NSCLC; and (3) reveal metabolic bypass as a potential mechanism of therapy resistance.

NIH Research Projects · FY 2025 · 2020-01

Abstract The life cycle of Mycobacterium tuberculosis (Mtb) is complex, encompassing an acute phase, during which the pathogen replicates exponentially; a chronic phase, when bacterial burden is stably maintained, and a latent paucibacillary state that can reactivate. Chronic tuberculosis (TB) is associated with the development of tissue-damaging immunopathology and can promote disease transmission. It has been estimated that approximately 1/4 of the world's population are infected with Mtb, and a significant proportion of these individuals harbor latent bacilli that can reactivate to cause diseases. Unraveling the mechanisms that regulate Mtb growth in an infected host in the different phases of infection is paramount to understanding TB pathogenesis. It is generally thought that certain host environmental conditions (e.g., hypoxia, nitrosative stress, starvation) can promote the establishment of a latent infection. However, the precise mechanisms that regulate TB latency are incompletely defined. Mtb Rv2623, which is among the most upregulated genes in the dormancy regulon, encodes a universal stress protein (USP) that can regulate bacillary growth both in vivo and in vitro. A deletion mutant ΔRv2623 is hypervirulent in susceptible mice and Guinea pigs, and in the latter, it is defective in establishing a chronic persistent infection. In vitro, overexpression of Rv2623 in mycobacteria retards growth in recipient cells; and Mtb ΔRv2623 exits from the non-replicative phase of the hypoxia-induced Wayne latency model more expeditiously than wild-type (WT) Mtb upon transfer into O2-sufficient media. These results provide evidence that Rv2623 regulates Mtb growth, including possibly during the latent/reactivation phase of infection. We showed that Rv2623 interacts with the FHA domain-containing Mtb Rv1747, a putative exporter of lipooligosaccharides. The FHA domain is a signaling protein module that mediates a wide variety of biological processes via phosphorylation-dependent mechanisms. We further showed that the Rv2623-Rv1747 interaction is mediated through binding of the FHAI domain of Rv1747 with a phosphothreonine (at position 237)-containing motif of Rv2623, and that the T237 residue is essential for mediating the growth-regulatory attribute of Rv2623. In contrast to the hypervirulent ΔRv2623, ΔRv1747 is attenuated for growth in vivo. And while the hypervirulent ΔRv2623 expresses enhanced levels of the immunoregulatory phosphatidyl-myo- inositol mannosides (PIMs) relative to WT Mtb, the hypovirulent ΔRv1747 is a hypo-producer of PIMs. In addition, we showed that Rv1747-overexpressing strains hyperproduce PIMs. The correlation of Rv1747's expression levels and Mtb cell wall PIMs amounts suggests that Rv1747 may function as an exporter of Mtb cell wall biogenesis intermediates. This, together with the opposing PIMs phenotype and in vivo growth phenotype of ΔRv2623 and ΔRv1747, has led us to hypothesize that Rv2623 negatively regulates the functional activity of Rv1747 to modulate the levels of Mtb cell wall PIMs, which immunoregulatory properties can alter Mtb-host interactions, thereby influencing the in vivo fate of the tubercle bacillus. We will use biochemical, genetics, and immunological approaches, in conjunction with animal modeling and integrative bioinformatics and computational data analysis, to rigorously test this hypothesis. Finally, accumulating knowledge derived from functional and structural analysis of Rv1747, and the discovery of the relationship between Rv1747 expression and PIM levels, will enable the generation of a set of isogenic Mtb mutants expressing graded levels of PIMs, which can be used to stringently probe the significance of these immunoregulatory glycoplids in influencing the in vivo fate of Mtb. The proposed studies should illuminate how the Rv2623-Rv1747-PIM pathway regulates in vivo Mtb growth. The data generated may help gain insight into the function of Rv1747 in modulating the cell wall PIM levels, the roles of PIMs in impacting the fate of the tubercle bacillus in an infected host, Mtb cell wall biogenesis, and potentially the mechanisms that regulate TB latency and reactivation.

NIH Research Projects · FY 2026 · 2019-05

Project Summary Over 250 million courses of antibiotics are prescribed annually to out-patients in the United States. The perception that antibiotic use has minimal adverse side effects contributes to their over-use. Since deploying antibiotics seems to be relatively free of toxicity, there are few disincentives to their use even when perceived benefits are marginal. Recent understanding of the human microbiome suggests the relevance of antibiotic exposures to health, with the emerging view that our microbiota are central to human physiology. In animal models, perturbing the microbiota affects metabolic, immune, and cognitive physiology. Antibiotics diffuse into all body compartments, selecting for resistance. We propose to examine the effects of two commonly used antibiotics, a beta-lactam (amoxicillin)and a macrolide (azithromycin) on microbial populations and on metabolic and immune physiology, in healthy volunteers in a prospective randomized clinical trial conducted at the NIH Clinical Center (CC). We hypothesize that in addition to acutely perturbing the human microbiome, these agents will have measurable metabolic and immunologic effects, with effects continuing over the following weeks and months. To test this hypothesis, in Aim 1, we assess the effects of a brief therapeutic antibiotic course on microbiota and metagenome composition. After initial evaluation, antibiotics are given for 5-7 days, with a prolonged post-treatment evaluation. Specimens are obtained from multiple sites at each of 10 time-points and used to estimate bacterial and fungal composition and gene content. In Aim 2, we assess the effects of the antibiotic course on immune physiology. At multiple time points, blood is obtained and used to determine plasma and cellular levels of markers of both innate and adaptive immunity. In Aim 3, we assess the effects of the antibiotic course on metabolic physiology. The specimens obtained are assessed for markers of metabolic and hormonal physiology. A subset of subjects enter the unique CC Metabolic Chamber to quantify 24-hour energy expenditure and carbohydrate and fat utilization. In addition to the primary data analyses, we will build an informatic model integrating the temporal data to provide insight into the complex interdependent physiology between microbiome and host. This project is an opportunity to perform comprehensive and integrated evaluations of two pharmacologic agents prescribed >90 million times annually in the USA. Careful analysis and development of an integrated model to understand the pathophysiology of the perturbations may identify problems below the radar in clinical medicine. The purpose of this renewal is to recruit sufficient numbers of subjects to complete the study after the interruption caused by COVID-19 and the subsequent COVID studies in the Clinical Center. Increasing the number of subjects to ~20 per group will permit robust analyses of the extent and duration of microbiome changes due to the antibiotic perturbations, and their consequent effects on host metabolic and immune functionalities, and their interactions; the present limitations in numbers of participants constrain a thorough analysis.

NIH Research Projects · FY 2024 · 2019-05

Project Summary: Neural computation requires the coordinated effort of thousands of interrelated and often genetically similar neurons. These neurons form physically intermingled networks and subnetworks that act together to amplify and strengthen sensory perceptions or select motor action. Such co-active ensembles are known to be preferentially interconnected, and may represent a functional element of neural processing with unique properties, such as pattern completion and competition between ensembles. In this proposal I will gain a mechanistic understanding of how ensembles of co-active neurons interact by probing the function of individual and groups of neurons in an awake mouse. I will examine: how ensembles of pyramidal cells interact with other pyramidal cells and local inhibitory neurons in a visual task, how these ensembles influence motor behavior, and how specific ensembles respond to information from other cortical areas. Despite the potential importance of ensembles in cortical coding, the intermixed nature of these groups has made them particularly hard to study. While conventional optogenetic techniques can manipulate genetically identified neurons in a region, they are incapable of selectively manipulating intermingled neurons that differ only by their functional properties. Critically, new multiphoton optogenetic techniques are beginning to allow manipulation of cells chosen by their activity alone, however such techniques require further development. In the K99 phase of this proposal, I will continue my training through the development of novel optical systems for multiphoton stimulation and through use of these technique understand cortical function. By combining these new optical techniques with novel opsins designed for in vivo multiphoton use that I have already developed, I now have the ability to write in or edit neural activity across many neurons with a precision never before possible. By altering ensemble activity during visual perception I will determine the causal contributions of individual neurons as well as populations of neurons to sensory coding. In the R00 phase, through manipulations in motor cortex I will unravel the behavioral impact of these groups, probing the role motor ensembles play in motor action, and study how neurons interact across modalities. The ability to both edit and monitor the activity of neural subnetworks is critical to gaining a mechanistic understanding of perception and action. The conclusions we draw from this proposal will help to describe how all information is presented in the cortex, but can only be reached with advanced techniques.

NIH Research Projects · FY 2026 · 2019-05

This project will characterize a large family of transcription factors that regulate utilization of complex dietary and host glycans in Bacteroides, a major component of the human gut microbiota. Bacteroides specialize in utilization of complex carbohydrates found in dietary fiber and the mucous that lines the gut, converting the glycans into nutrients used by themselves, other bacteria, and humans. Bacteroides devote ~20% of their genomes to genes encoding proteins necessary for capturing, processing, importing and metabolizing glycans, with genes for utilization of specific glycans clustered into dozens of different polysaccharide utilization loci (PULs). Regulation of PULs determines which glycans are utilized, shaping the composition of the gut microbiota and the balance between diet and host glycan utilization, with impacts on human health including immunomodulation, neurotransmitter release and bowel inflammation. Hybrid Two-Component System (HTCS) transcription factors are a prevalent strategy used for regulating PULs. HTCSs are an unusual variation of Two-Component System (TCS) signaling in which the transmembrane sensor histidine kinase and response regulator transcription factor are combined into a single protein chain. Despite the critical role these regulators play in a central genus of the human gut microbiota, remarkably little is known about their function. Specifically, the genes regulated by each HTCS have not been identified, the regulatory network that produces prioritized utilization of different complex glycans has not been defined, and the mechanism of coordination of HTCSs that enables regulation of multiple PULs necessary for utilization of a single complex dietary fiber is unknown. Several obstacles have hindered investigations to date. The specific activating ligands of many HTCSs are unknown; DNA-binding sites for HTCS transcription factors are poorly defined; and direct vs. indirect effects on gene expression in response to specific glycans cannot be distinguished. Our recent studies have revealed a conserved feature of HTCSs that can be exploited to activate DNA binding independent of an activating ligand. This strategy will be employed to define the regulons of each HTCS using CHiP-seq and RNA-seq to identify DNA-binding motifs and establish regulatory sites for activation or repression. These data will enable identification of cross-regulation of multiple PULs by individual HTCSs, thus establishing a regulatory network that will provide insights into prioritization of glycan utilization. Mechanisms of coordination of HTCSs involved in pectin utilization will be explored using regulon analyses, HTCS interaction assays, and HTCS localization studies. Structural characterization of HTCSs by x-ray crystallography and cryoEM will be pursued to provide a comprehensive structural description of phosphotransfer signaling from ligand-binding input to DNA-binding output, furthering understanding of TCS signaling and elucidating features specific to HTCSs adapted for regulation of polysaccharide utilization. These studies will provide foundational knowledge about complex carbohydrate utilization by Bacteroides as a step toward developing dietary strategies to improve gut health.

NIH Research Projects · FY 2025 · 2019-03

ABSTRACT Invasive fungal infections are emerging diseases that kill over 1.5 million people each year. Among them Cryptococcus neoformans is the leading cause of fungal meningitis that is account for 15% of AIDS-related deaths. The treatment option for fungal infections is very limited and there is no fungal vaccine available in clinical use. Therefore, there is an unmet need to develop new treatments against this life-threatening fungal infection. Our recent studies uncovered that Fbp1, a F-box protein in the ubiquitin-mediated proteolytic pathway, acts as a master regulator of C. neoformans immunogenicity and identified fbp1 mutant as a new vaccine candidate. We found that vaccination with heat-killed fbp1 (HK-fbp1) conferred significant protection from a subsequent challenge with not only virulent parental strain H99 in mice, but also showed cross protection against C. gattii and other fungal infections, including Aspergillus fumigatus. Treating C. neoformans infected animals with HK- fbp1 vaccine also showed protection. Thus, HK-fbp1 is an optimal vaccine candidate with broad protection, and a better understanding of how HK-fbp1 induces protection will facilitate the future development of new treatments against cryptococcosis and other fungal infections. During the last funding period, we demonstrated that the HK-fbp1 vaccine protection is dependent on the activation of innate and adaptive immune responses that included the rapid recruitment and maturation of CCR2+ monocytes and the enhanced activation of CD8 and CD4 Th1/Th17 responses. Such protection is dependent on IFN- We also identified an Fbp1 E3 ligase substrate Crk1 that is critical for cell size regulation and Fbp1 mediated immunogenicity, leading to a better mechanistic insight. In ongoing studies, we determined that the addition of CpG oligodeoxynucleotides (CpG- ODN) to HK-fbp1 leads to improved vaccine efficacy. Importantly, HK-fbp1+CpG-ODN induced protection against Cryptococcus infection after intramuscular (i.m.) administration whereas i.m. HK-fbp1 alone is unable to protect. The immune mechanisms of this substantially improved protection mediated by the adjuvanted vaccine remain unknown. Based on our published and unpublished observations we hypothesize that the success of HK-fbp1 as an effective vaccine is critically linked to the enhanced capacity of fbp1 to induce protective innate and adaptive immune responses. Our overarching goal is to systematically decipher the immune mechanisms of vaccine-induced protection and to define the function of specific Fbp1-regulated targets that shape the immunogenicity of C. neoformans to develop a vaccine applicable for human use in the future. To test our hypothesis and achieve our goal, we propose to 1) decipher the distinct contributions of innate and adaptive immunity from adjuvanted HK-fbp1 vaccine-induced protection, and 2) define immunogenic factors in HK-fbp1 to develop an improved vaccine against fungal infections. These studies will advance our understanding of host- pathogen interactions involved in the immune regulation by Cryptococcus and guide the design of effective vaccines critical for the control of fungal infections.

NIH Research Projects · FY 2024 · 2018-09

ABSTRACT (No change from original, funded submission) Young adults’ initiation and use of e-cigarettes are on the rise in the U.S. E-cigarette use produces toxicants, is addictive, and is associated with future use of combustible tobacco products among young adults. The situation is compounded by aggressive e-cigarette marketing, which often features flavors, models, marketing claims, and price promotions. Evidence has shown the strong influence of tobacco marketing on the initiation and use of tobacco products among young adults, especially young adults who are naïve to tobacco. Thus, a timely public health response comprising more regulation on e-cigarette marketing is needed to prevent and reduce e-cigarette uptake among this group. Although e-cigarette marketing has been pervasive and is rapidly growing, there is a lack of studies examining the influence of e-cigarette advertisement features on young adults’ reactions and other behavioral antecedents. The proposed project will examine the influence of four e- cigarette advertisement features (flavors, models, marketing claims, and price promotions) among young adult non-tobacco users (including never users and experimenters) who are susceptible to e-cigarette use. This study will pursue two Specific Aims: Aim 1 is to identify key features of e-cigarette advertisements that lead to greater attention, cognitive arousal, and positive emotional responses; and Aim 2 is to determine whether edited e-cigarette advertisements without key features lead to reduced positive e-cigarette perceptions and behavioral intentions compared to original, unaltered advertisements. The Aim 1 study will use within-subjects design and adopt eye-tracking technology equipped with pupil diameter assessment and facial expression analysis among 70 young adults. The Aim 2 study will adopt a between-subjects comparative randomized experiment among 900 nationally representative young adults enrolled in an online panel. Individual differences (e.g., biological sex and tobacco use history) in neurocognitive reactions and e-cigarette related perceptions and behavioral intentions will also be assessed for the two studies. The proposed research will innovatively use neuroscience technologies to objectively measure young adults’ neurocognitive reactions to e- cigarette advertisements and help draw causal inferences between viewing e-cigarette advertisements features and immediate perceptions and intentions of using e-cigarettes. This research is directly relevant to the development of the FDA’s policymaking efforts to reduce the impact of e-cigarette advertisements on potential uptake of e-cigarettes among young adults who are naïve to tobacco products.

NIH Research Projects · FY 2025 · 2018-09

PROJECT SUMMARY Glutamine (Gln) is a multifaceted amino acid that serves as important carbon and nitrogen sources. In a situation where circulatory supply of Gln is limited, such as in poorly vascularized tumors, cell autonomous Gln synthesis is turned on to assimilate the inorganic ammonia into Gln to support the nitrogen anabolic processes. In mammals, Gln synthesis is catalyzed by glutamine synthetase (GS) that condenses ammonium with glutamate to produce Gln. GS has been shown to enhance cell survival/growth/repairs in various cancers by promoting de novo glutamine synthesis and subsequent nitrogen anabolism. In the liver, where blood supply of Gln is abundant, GS-mediated Gln synthesis serves as a mechanism for ammonia detoxification next to the urea cycle. Interestingly, in hepatocellular carcinoma (HCC), the major histological subtype of liver malignancy, GS expression is frequently elevated whereas urea cycle enzymes (UCEs) down-regulated. Both elevated GS and decreased UCEs correlate well with -catenin activation, a prevalent oncogenic driving event in HCC. While this seems to be consistent with the common thought that GS promotes tumor growth, surprisingly, using the Glulflox/flox mouse, we recently reported that genetic ablation of hepatic GS accelerated tumor growth in several HCC models that involve -catenin6. Echoing the scenario in clinic, oncogenic -catenin suppressed the expression of the UCEs while induced the expression of GS. It was then speculated that the suppression of UCEs led to defective ammonia clearance, and GS was upregulated to help alleviate the hyperammonemia situation. GS ablation exacerbated the ammonia burden and facilitated the production of Glu-derived non- essential amino acids (NEAAs) that subsequently stimulated mTORC1. These findings prompt us to form the hypothesis that GS-mediated ammonia clearance functions to suppress tumor growth in -catenin-driven HCC. We propose two Specific Aims: 1) Study the theory that defective ammonia clearance promotes HCC. We will first determine whether GS suppresses HCC growth in a cell-intrinsic fashion, study how hyperammonemia promotes HCC, and take unbiased approach to uncover other mechanisms that may be responsible for enhanced HCC growth resulting from hyperammonemia. 2) Study how -catenin regulates GS expression in the liver. GS transcription is activated by Wnt/-catenin during liver zonal development and upon oncogenic transformation of hepatocytes. However, the underlying mechanism remains elusive. Our preliminary data suggest that GS expression is regulated by a liver-specific cis-acting distal enhancer. We will identify and characterize the liver-specific distal enhancer and study the underlying mechanism for GS expression regulated by the novel transcription factors that are associated with the enhancer. Accomplishing these Aims will help establish a novel theory that GS-mediated ammonia clearance is a tumor-suppressing mechanism in HCC, and will uncover the molecular basis for liver-specific regulation of GS expression driven by -catenin.

NIH Research Projects · FY 2026 · 2018-07

ABSTRACT Increasing the retail age of sales of commercial tobacco products from 18 to 21 (known as Tobacco 21 or T21) has been shown to decrease youth and young adult tobacco access and use. However, underage sales are widespread despite the enactment of T21, and retailer characteristics may contribute to underage sales rates. In addition, the tobacco retail and regulatory environment has changed considerably since federal T21 became effective in December 2019 (e.g., temporary suspension of T21 enforcement during COVID, tobacco product diversity, regulatory actions, electronic identification [ID] verification technology). Using the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) framework and Purtle’s conceptualization framework of policy in implementation science research, we seek to better understand compliance and enforcement of T21 over time and how T21 implementation can be improved, including identifying modifiable factors that affect key implementation outcomes (i.e., ID check and sales to underage purchasers) and may lead to differential implementation outcomes across social groups and neighborhoods. This competing renewal application intends to examine how tobacco product types, retailer characteristics, and neighborhood-level factors, contribute to T21 implementation in a longitudinal sample of tobacco retailers in three states: New Jersey, the focus of our original application; North Carolina, a state added after federal T21; and now Nevada, the first state to require electronic ID verification. Specifically, by conducting underage purchase attempts in a longitudinal sample of retailers (at least 4 Waves of data collection), we aim to 1) Examine T21 implementation (i.e., ID checks, sales to underaged buyers) over time by tobacco product types to determine the propensity of tobacco retailers to check IDs and sell cigarettes, cigars, electronic cigarettes, smokeless tobacco, and nicotine pouches to underage purchasers; 2) Identify the extent to which retailer-level characteristics (i.e., store type, tobacco industry contracts, electronic ID verification) contribute to tobacco product ID checks and underage sales over time; 3a) Determine how neighborhood-level factors (i.e., sociodemographic characteristics, tobacco retailer availability) are associated with tobacco product ID checks and underage sales over time and finally, 3b) Test whether neighborhood sociodemographic characteristics and tobacco retailer availability moderate associations between tobacco product types (Aim 1) and retailer-level characteristics (Aim 2) and tobacco product ID checks and underage sales. Results from this project will provide a longitudinal evaluation of T21 implementation and identify modifiable characteristics that can inform regulatory efforts, retailer education programs, and overall T21 compliance and enforcement.

NIH Research Projects · FY 2025 · 2018-05

Project Summary/Abstract Combining high-throughput biomedical data sets from multiple studies is advantageous to increase statistical power in studies where logistical considerations restrict sample size or require the sequential generation of data. However, significant technical heterogeneity is commonly observed across multiple batches of data that are generated from different processing or reagent batches, experimenters, protocols, or profiling platforms. These so-called batch effects confound true relationships in the data, reducing the power benefits of combining multiple batches of data, and may even lead to spurious results. Many methods have been proposed to filter technical heterogeneity from genomic data. These methods are designed to remove batch effects, unmeasured or “surrogate” variation, or other “unwanted” variation caused by biological or technical sources. Although these approaches represent impactful advances in the field, there are still significant gaps that need to be addressed to appropriately filter technical heterogeneity from -omics data and other high-throughput datasets. For example, many existing methods assume relevant covariates are known or that raw data are generally independent. Some applications require more specific and direct correction methods, including single cell transcriptomics data that are often missing cell-type identifiers, microbiome data that are compositional in nature, and imaging and spatial transcriptomics data that have spatially correlated data points. Furthermore, batch correction introduces correlation into the adjusted data, which needs to be accounted for in downstream analyses, and most researchers performing batch correction are unaware of this negative impact and often incorrectly apply downstream analysis tools. Finally, there is still significant need for additional software tools and benchmark datasets for evaluating batch effect methods and their efficacy in specific datasets. We propose to develop algorithms and software to address these specific research gaps facing researchers combining data from multiple experimental batches.

NIH Research Projects · FY 2026 · 2018-02

Project Summary: This proposal is a renewal application of R01 (AI130197). In the previous funding cycle, we sought to identify the mechanism(s) by which the small adaptor protein Crk controls NK cell activation and inhibition, using human NK cells and NK cell-specific conditional knockout mice. We found that Crk phosphorylation plays a key role in function and that Crk haploinsufficiency leads to NK cell deficits and impaired integrin signaling. Importantly, we identified new small molecule inhibitors of pCrk (Patent #: 63/305,354). In the proposed renewal, we use this new tool to further dissect Crk’s ability to integrate activating and inhibitory input at the immune synapse (IS) of both NK and T cells in a variety of relevant settings: in high-resolution live human primary cell imaging assays, in human Crk haploinsufficiency, and in triple negative breast cancer (TNBC). We test our central hypothesis that Crk functions as a master regulatory switch, fine-tuning activating and inhibitory influences via interactions with downstream molecules and the actin cytoskeleton, and propose the new hypothesis that pCrk inhibition can restore exhausted immune responses in cancer via novel signaling pathways. Guided by strong preliminary data, we propose three Specific Aims: 1) How does Crk function as a master regulatory switch to control NK cell function? Here, we use cutting-edge imaging systems, lipid bilayers, live human primary cells, RNA-Seq, and our new CrkL inhibitors to determine where Crk, pCrk, and their upstream and downstream partners localize in the cytotoxic, inhibitory, and exhausted IS, and how pCrk inhibitors influence NK signaling in healthy and exhausted NK cells. 2) How does Crk influence immune function and integrin signaling in pDGS T cells? Our previous program showed that NK cells from patients with pDGS – a common genetic syndrome resulting from haploinsufficiency of CrkL-containing locus 22q11 – exhibit functional NK cell deficiencies, as well as defective integrin activation at the IS. We expand these studies to T cells, using primary pDGS patient cells and integrin conformation-specific antibodies to assess how Crk functions as a two-way regulatory switch for CTL function. We present data that haploinsufficiency of CRKL results in functional T cell deficits, and examine integrin signaling, T cell function, and IS quality with an eye towards clinical application of small molecule LFA-1 agonists to restore pDGS-related functional immune cell deficiencies. 3) Discover the role of pCrkL inhibitors in cancer immunotherapy. Our preliminary data show that pCrk inhibitors significantly inhibit tumor growth, restore cytotoxicity of inhibitor receptor-dominant immune cells, and enhance survival. We hypothesize that pCrk inhibitor-mediated solid tumor suppression results from both direct tumor killing and restoration of exhausted immune cell function. In Aim 3, we use a battery of TNBC mouse models to elucidate Crk’s molecular mechanisms in solid tumors, testing the impact of Crk inhibition on tumor growth and cell death, immune cell function in the tumor microenvironment, and metastasis. Our long-term goal is to develop new treatments for human immunodeficiencies and cancer.

NIH Research Projects · FY 2026 · 2018-01

Project Summary/Abstract The current proposal is to support a Summer Undergraduate Research Program in Neuroscience (NeuroSURP) mentored by Rutgers University research faculty. The program has three primary aims: (1) To increase student knowledge and appreciation of basic biological research by providing a closely-mentored, hands-on research experience; (2) To increase student knowledge and interest in pursuing careers in research through career development and educational activities; (3) To provide continued advice, support and guidance to program alumni to facilitate post-program career planning and implementation. Twelve undergraduates will be admitted to our program each year beginning 2024. Special consideration will be given to students from historically underrepresented minority and disadvantaged backgrounds including Black, Hispanic, Native American, Pacific Islander, or have physical or mental disabilities, or are from economically and/or socially disadvantaged backgrounds (collectively referred to as URM) that comprise a deep and diverse pool of potential future scientists. The research component of the proposed SURP involves hands-on experimentation in labs with active, nationally recognized and funded research programs in neuroscience with a focus on neurological disorders at Rutgers University. The infrastructure and general organization of the proposed program will be based on an existing program that has been in place for 27 years. Students will work on their own research projects under the close supervision of mentors in their host labs. The proposed SURP is designed to integrate student research experiences and mentoring with weekly meetings comprising training in neuroscience, oral and written presentations, critical reading of manuscripts, Responsible Conduct of Research as well as Rigor and Reproducibility including statistical analysis. In addition, there is a significant career development component during the weekly meetings in which PhD, MD/PhD and MD guest speakers discuss their career paths representing academic research, non-academic research as well as research- related careers. Each year students will present the results of their summer research projects in a symposium that is open to scientists on campus and members of the general public. In addition, the students submit a paper on their project in the format of a manuscript and receive feedback from MPIs. Finally, the Co-directors will maintain close contact with the alumni of our program as we have done for the past 27 years and will continue to provide them with advice and mentoring related to applying to graduate school and their careers. The mentors who will be supervising the students study neurological disorders including Alzheimer's, Parkinson's, multiple sclerosis, autism, traumatic brain injury, spinal cord injury, sensory loss, epilepsy, and stroke. The multidisciplinary research focus of the faculty mentors on the cellular, molecular, and genetic processes underlying developmental and degeneration of the nervous system has implications for the etiology, pathogenesis and progression of neurological disorders.

NIH Research Projects · FY 2026 · 2017-09

ABSTRACT With the recent revolution in artificial intelligence for molecular design and protein structure prediction, we were motivated to apply these new tools to address bottlenecks in our research focusing on retroviral entry, integration, and drug design. Viral entry and receptor usage has become a major focus with the emergence of new viruses with expanded host and tissue specificities. Our research focuses on the ability of retroviral Envelopes to select and utilize novel host cell receptors. In this proposal, we are utilizing new approaches to identify and study two novel retroviral Env proteins and their host cell receptors. These include the binding of the A5/A9a Env protein to the host membrane protein SLC35F2 and the identification of the cognate receptor(s) for the L1 Env. Applying AlphaFold2, Phyre2 and Evolutionary Covariance structure prediction programs, we have a first generation model for SLC35F2, which serves as the foundation for a broad number of questions relating to the function of this orphan receptor. The second area of focus builds on the gammaretroviral Integrase (IN) protein binding to the host BET proteins. Through our structural studies of both the Murine Leukemia Leukemia (MLV) IN protein and the host NSD3 protein binding to the ET domain of the Brd3 BET protein, we are now able to apply our knowledge of the common binding pocket to advance approaches for studies of alternative binding substrates and small molecule inhibitors. The ET domain of the BET family of proteins serves as a node for multi-protein assemblies. Understanding the affinity of different proteins and small molecules to this ET binding pocket can have profound effects of gammaretroviral pathogenesis as well as gene and oncogene regulation. Using computer-aided drug discovery approaches, we have identified first-in-class lead compounds targeting the ET domain. These studies build on our cohesive collaborative research team with expertise in virology, NMR structural studies, and bioinformatics.

NIH Research Projects · FY 2026 · 2017-09

Project Summary Porphyrias are genetic disorders caused by mutations in enzymes involved in eight sequential biosynthetic conversions that combine glycine and succinyl coenzyme-A in the first enzymatic step to ultimately generate heme. Porphyrin accumulation also occurs in ‘secondary porphyrias’ in association with other diseases such as hepatitis C virus infection. Current major unmet needs with regard to the porphyria disorders include: (a) our present limited understanding of the biochemical mechanism of cell and tissue injury, (b) the molecular triggers of porphyria acute attacks, (c) the reasons why some individuals develop significant organ complications such as end-stage liver disease that requires liver or bone marrow transplantation, while others do not, and (d) the limited availability of drugs to treat the different porphyrias. Our central hypothesis is that the liver is susceptible to light-independent porphyrin-mediated proteotoxic damage that leads to cell and tissue injury in porphyria, and that drugs can be identified that lead to increased or decreased porphyrin accumulation. This hypothesis will be tested by pursuing three interconnected specific aims: (i) Define the mechanism of light-independent porphyrin- induced protein aggregation in internal organs, with a focus on the liver; (ii) Elucidate the mechanism of detoxification of porphyrin-induced proteotoxic damage using in vitro and in vivo models; and (iii) Characterize small molecules that decrease or increase tissue porphyrin accumulation and porphyrin-mediated proteotoxicity. We have assembled extensive preliminary results to support the likely success of our aims, including substantial evidence for porphyrin-mediated protein aggregation that is light-independent, the reversibility of protein aggregation and enzymes that are likely to be involved in reversing protein oxidation, and the use of zebrafish high-throughput screening to identify known drugs that decrease or increase porphyrin accumulation in liver. The drugs that decrease porphyrin accumulation will be tested for their mechanism of action and examined in preclinical porphyria experimental models as drugs that may be repurposed as potential new therapies. In parallel, drugs that increase porphyrin accumulation will be characterized as potential candidate drugs to avoid in patients with porphyria. Completion of our proposed aims provides fundamental knowledge regarding which proteins are prone to porphyrin-mediated oxidation and aggregation, the molecular signatures that define such aggregation, the mechanism of aggregate turnover and disaggregation, whether compounds we characterize are candidates for testing in patients with porphyria, and whether currently used drugs in non-porphyria disorders might need to be avoided or monitored in patients with porphyria. This proposal uses state-of-the-art technologies, multiple biochemical and porphyria animal model tools including zebrafish and mice, and introduces the novel concept of proteotoxicity as an alternative mechanism for porphyria exacerbations and progression that may shed light on genetic modifiers that account for liver disease progression in some patients but not others.