Rutgers Biomedical And Health Sciences

NIH Research Projects · FY 2026 · 2017-08

PROJECT SUMMARY Congenital heart disease (CHD) is the most common birth defect, affecting approximately 1% of newborns. About 25% of CHD patients require surgery within the first year of life and often need lifelong medical care. Despite advances in surgical care, CHD mortality remains high, highlighting the need for a better understanding of its underlying causes. Abnormal development of the arterial pole of the heart accounts for roughly 30% of CHD cases, leading to severe and often lethal malformations due to the misalignment of the great arteries with the ventricles. The great arteries arise from the cardiac outflow tract (OFT). Initially, the OFT forms as a single vessel connecting the heart with systemic circulation. As the embryo develops, the OFT elongates and becomes subdivided into two vessels, the aorta and pulmonary artery, which then rotate to align with the left and right ventricles, respectively. However, if the OFT fails to elongate during its initial stages of formation, the great arteries fail to properly align with the ventricles, resulting in the mixing of oxygenated and deoxygenated blood after birth. This causes severe morbidity and lethality in the absence of surgical intervention. The OFT elongates via the addition of progenitors from the second heart field (SHF). SHF-derived cells form an epithelial layer in the dorsal pericardial wall (DPW). OFT elongation depends on both tissue tension and epithelial organization in the DPW. When either is altered, SHF cells fail to migrate and incorporate into the OFT, causing defective OFT elongation and, consequently, aberrant great artery morphogenesis. We discovered that the conditional ablation of fibronectin (Fn1) in the cardiogenic mesoderm phenocopies the SHF and OFT abnormalities observed in Tbx1-null mutants, a model of 22q11.2 deletion syndrome—the most prevalent chromosomal disorder in humans. Like Tbx1, Fn1 controls OFT elongation by regulating epithelial organization and mechanotransduction, specifically in the anterior DPW. In this grant application, we propose to determine the mechanisms by which Fn1 regulates epithelial cell shape and mechanotransduction in the SHF. We will test the hypothesis, supported by our preliminary data, that Fn1 regulates SHF cell architecture, biomechanical properties, and OFT elongation by balancing cell- extracellular matrix (ECM) interactions with the anti-adhesive ECM glycoprotein Tenascin C (TnC). We hypothesize that balanced interactions of SHF cells with Fn1 and TnC are required for SHF cells to achieve proper cell shape, size, polarity, and nuclear enrichment of YAP, all of which are necessary for OFT elongation and great artery morphogenesis. Upon completion of these studies, we will gain new insights into the molecular and genetic regulation of OFT elongation and morphogenesis of the arterial pole of the heart. Our research will provide novel candidates (Fn1 and TnC) for prenatal CHD screening and inform targeted therapeutic approaches in the future. Ultimately, understanding how alterations in ECM composition contribute to the pathogenesis of CHD promises to broaden therapeutic strategies and enhance patient outcomes.

NIH Research Projects · FY 2025 · 2017-08

Project Summary T helper type 2 (Th2) cells play a crucial role in allergies, humoral immunity and host protection against parasitic infections, but our understanding of the mechanism of their differentiation remains incomplete. Upon encountering a cognate antigen presented by dendritic cells (DCs), naive CD4T cells make a fate decision to become one of the effector cell types such as Th1, Th2, and Th17 cells. However, unlike their differentiation into Th1 or Th17 cells, in which IL-12 and IL-23 play a crucial role, respectively, the DC-derived fate instruction signal universally required for Th2 differentiation has not been identified. CD301b (Mgl2) and its human homolog CLEC10A are selectively expressed by a major subset of type 2 conventional DCs in the dermis and other peripheral organs. We previously developed mouse models in which CD301b+ DCs are depleted, enriched or genetically manipulated and demonstrated that CD301b+DCs are both necessary and sufficient for the Th2 cell differentiation upon exposure to a protease allergen papain in the skin as well as after infection with a hookworm Nippostrongylus brasiliensis. Interestingly, the depletion of CD301b+DCs results in a shift of the effector CD4T cell phonotype from Th2 to Th1 and Th17 phenotype, suggesting that CD301b+ DCs regulate the effector cell fate at the clonal level. The molecular mechanism for the Th2 fate instruction by CD301b+DCs still remains unclear, but our data indicate the requirement of cognate interactions between CD301b+DCs and antigen-specific CD4T cells. Thus, we hypothesize that CD301b+ DCs imprint the Th2 fate on CD4T cells mainly through a contact-dependent, rather than soluble, mechanisms. In this application, we aim to address the following two questions by vigorously characterizing the CD4T cell differentiation kinetics in our unique mouse models: (1) What is the CD4T cell-intrinsic, Th2-skewing signal(s) imprinted by CD301b+ DCs?; and (2) How do CD301b+ DCs instruct antigen-specific CD4T cells to become Th2 cells? Answering these questions will deepen our basic understanding of the in vivo mechanism of the initiation of a Th2 response and help us to develop a unified model that comprehensively explains the role of DC subsets in CD4T cell differentiation. Understanding such mechanism would help us to improve our strategies for treating allergic diseases and developing effective vaccines for infectious diseases.

NIH Research Projects · FY 2026 · 2017-07

Project Description Vitamin D is an essential nutrient whose active hormonal form, 1,25-dihydroxyvitamin D (1,25(OH)2D), regulates vitamin D receptor (VDR)-mediated gene expression to stimulate intestinal calcium absorption, to maintain intestinal barrier function, to suppress colonic inflammation and to suppress carcinogenesis. However, intestinal resistance to vitamin D develops with advancing age. This reduces intestinal Ca absorption in rodents and in humans but it is not clear how age-associated intestinal vitamin D resistance affects other aspects of intestinal vitamin D action. Thus, there is a critical knowledge gap regarding the mechanisms for how intestinal 1,25(OH)2D action changes across the lifespan. Our long-term goal is to determine how vitamin D regulates intestinal biology across the lifespan to modulate classical (Ca absorption) and non-classical (cancer, inflammation) vitamin D endpoints. Our studies from the past grant period were the first to show the complexity of 1,25(OH)2D- regulated gene regulation across the functional compartments of the intestine (i.e. small intestine (SI) crypt, SI villus, colon). Our new preliminary data demonstrate that advanced age can suppress the intestinal induction of some, but not all, vitamin D target genes. Thus, we hypothesize that age-related intestinal resistance to 1,25(OH)2D is due to context-dependent interference of VDR transcriptional mechanisms. To address this hypothesis, we have designed three aims: Aim 1: Determine how 1,25(OH)2D genomic action is modified across the lifespan to result in age-associated intestinal vitamin D resistance. Based on our pilot data, we hypothesize that the impact of aging on vitamin D-dependent gene expression is not uniform across genes or by intestinal compartment (i.e. SI/colon, crypt/villi). We will use state-of-the-art genomic approaches to study age- and compartment-specific changes in intestinal vitamin D action. Aim 2: Determine how VDR genomic action depends upon other transcription factors and co-regulators across the lifespan. We will conduct studies to reveal the critical, VDR-interacting proteins that define compartment- and age-sensitive regulation of intestinal gene expression and functionally test their roles in organoid models. Aim 3: Test whether targeting 1,25(OH)2D to the proximal colon can enhance Ca absorption and reduce age-related bone loss. We will determine whether colon-targeted forms of 1,25(OH)2D can enhance Ca absorption in mice to prevent trabecular bone loss in adults and increased cortical porosity in old mice. Our proposed studies provide the unique opportunity to determine (a) the scope of genes and regulatory regions impacted by aging in the intestine, (b) the role of intestine-specific transcription factors and co-activators in age-associated intestinal vitamin D resistance, and (c) whether colon-targeted forms of 1,25(OH)2D can have health benefits by overcoming age-associated vitamin D resistance and Ca malabsorption. When complete, this research will serve as a scientific foundation for developing strategies to improve intestinal vitamin D action, especially in older populations at high risk for developing diseases like osteoporosis as well as colon cancer, and inflammatory bowel syndromes.

NIH Research Projects · FY 2026 · 2016-11

ABSTRACT Cryptococcus neoformans (Cn) is the most common cause of fungal meningoencephalitis, a life-threatening disease. Despite extensive efforts, the mechanism whereby Cn crosses the blood-brain barrier (BBB) and causes meningitis remains poorly understood. Our previous study discovered that inositol, a host sugar metabolite highly abundant in the brain, promotes fungal traversal of the BBB and plays a critical role in host- pathogen interactions during infection of the central nervous system (CNS). In our previous funding period, we have demonstrated that Cn contains an unusually complex inositol uptake system that is distinct from those characterized in other fungi. Thus, Cn may be uniquely adapted to thrive in the inositol-rich environment of the CNS and to utilize inositol-dependent pathways for pathogenesis. Our data have confirmed that (1) growth of Cn under inositol-rich conditions enhanced virulence in the murine model, (2) fungal cells deficient in inositol uptake and catabolism were also deficient in virulence during CNS infection, (3) inositol induced production of fungal capsule, a major virulence factor, and hyaluronic acid, a cell surface molecule involved in fungal binding to the BBB, and (4) inositol induced a unique capsule structure enriched in a specific mannosylated structure group that helps the fungus evade the host immune response. We also identified an inositol receptor Itr4 and a transcription factor Hlh6 that are required for inositol function in Cn. Our central hypothesis is that cryptococcal cells sense and utilize host inositol to modify the fungal cell surface in a way that promotes penetration of the BBB and development of cryptococcal meningoencephalitis. The overarching goal of this proposal is to understand how this pathogen senses and utilizes host nutrient inositol to establish human brain infections. To further test our hypothesis, we propose two related but independent Specific Aims: 1) Characterize Inositol mediated fungal cell surface modification in Cryptococcus-host interaction using genetics, molecular biology and biophysical approaches in combination of in vitro BBB system and in vivo animal models. Fungal strains with single motif capsule structure will be utilized for this study. 2) Define the inositol sensing and regulatory mechanism important for fungal pathogenesis by characterizing the inositol sensor Itr4 and inositol transcription factors and the pathways they regulated. Moreover, we will test a number of inhibitors on inositol function for their potential antifungal activity to explore the possibility to develop the fungal inositol function as a potential therapeutic target. The proposed study will provide a mechanistic understanding of inositol utilization in Cn and its effects on fungal pathogenesis, especially during CNS infections.

NIH Research Projects · FY 2025 · 2016-05

Project Summary The overarching goal of our research is to understand the mechanisms of helicases and polymerases in processes such as viral RNA recognition, DNA transcription, and replication. The unifying approach is the quantitative characterization of the enzymatic reactions using rigorous biochemical and biophysical methods such as transient state kinetics, single-molecule kinetics, computational kinetic modeling, and cryo-electron microscopy. The integration of structural and functional studies allows the development of a complete mechanistic picture. In project 1, we are studying viral RNA recognition by RIG-I like receptors which are helicases serving as the first responders of viral RNA infections. The RIG-I like receptors recognize pathogen- associated molecular patterns on viral genomes and replication intermediates and respond by triggering an immune response to create an antiviral state. Our research focuses on understanding the mechanisms of RNA recognition and ATPase/helicase functions of RIG-I like receptors using biochemical, structural, and cell- signaling assays. We are elucidating the intrinsic mechanisms in RIG-I that enable self versus non-self recognition and developing new strategies to understand how they are activated and regulated. In project 2, we are studying the mechanism and regulation of mitochondrial DNA transcription catalyzed by RNA polymerases that resemble phage T7 but regulated by transcription factors. Transcription initiation and transition into elongation are key stages that are regulated by transcription factors. We are using cryo-electron microscopy, and ensemble/single-molecule kinetics to elucidate the structure and dynamics at these stages of transcription using in vitro reconstituted yeast and human mitochondrial RNA polymerases. In project 3, we are studying the mechanism of DNA replication by phage T7 and human mitochondrial replisomes. We study how helicase and polymerase work together to catalyze strand-displacement DNA synthesis, in particular, how they are energetically coupled. We are studying the mechanism of DNA synthesis by mitochondrial DNA polymerase to understand the role of helicase, Twinkle, and mitochondrial single-strand binding protein. An in- depth understanding of the enzymatic mechanisms is critically necessary to understand mitochondrial DNA deletions caused by defects in helicase and polymerase. This research will provide the mechanistic framework to quantitatively model the reactions of replication, transcription, and pathogen recognition that will guide in the development of therapies for viral infections, cancer, mitochondrial diseases.

NIH Research Projects · FY 2026 · 2016-02

Abstract and summary Alcohol use disorders (AUD) are complex behaviors accompanied by substantial morbidity, mortality and societal expense. Both genetic and environmental factors contribute to AUD. Despite progress in the human genetics of AUD, especially the identification of genome-wide significant AUD genetic risk variants, the neural basis of AUD in humans is largely unknown. Over the past five years, we have provided compelling evidence that: 1) Human neuronal cells derived from induced pluripotent stem (iPS) cells can be used as a tractable model to study neuropsychiatric disorders including AUD; 2) Ethanol exposure results in an inflammasome response in human neurons; 3) Human neurons carrying OPRM1 A118G minor gene variants showed enhanced sensitivity to opioids and ethanol; 4) Ethanol causes gene expression changes in both human neurons and glial cells; and 5) Microglia-containing 3D neural cultures can be a powerful system to study neuroinflammatory microglia-neuronal interactions. These premises provide a foundation for further mechanistic studies of the genetic and molecular underpinnings of AUD in human iPS cell-derived neural models. We also identified key gaps of knowledge in utilizing human neurons as a model system to study AUD that need to be filled, particularly: 1) how ethanol affects neuro-glial interactions in a human neural context is not known; and 2) how ethanol affects neurogenesis in a 3D context is not known. Moreover, 3) because of the polygenic nature of AUD, the contribution of single gene to AUD risk is likely small and the phenotypical manifestation is strongly influenced by individual’s genetic makeup. To address these outstanding questions, we hypothesize that ethanol impairs neuronal function via affecting neuroglial interactions, which is influenced by individual polygenic risk backgrounds. We have selected 36 subjects of both sexes with either extremely high polygenic risk score (PRS, top 10%ile, n=18 AUD) or low PRS (bottom 10%ile, n=18, no AUD) to test this hypothesis. We will differentiate iPS cells derived from these subjects into both 2D and 3D (i.e. brain organoids) neuronal cells co-cultured with human astrocytes and human microglia. Upon exposure to ethanol, these neural cells are subjected to a combination of morphological, immunocytochemical, electrophysiological, live cell imaging and genomic analyses to unravel the mechanism(s) underlying the impact of ethanol, focusing on neuro-astro-microglial interactions. In a relatively large collection of human iPS cells (n=36 lines) derived from subjects with extreme AUD PRSs, we hope to unravel the convergent phenotype and gene-networks that are linked to extreme high or low AUD PRSs. The results will advance our mechanistic understanding of the pathogenic role of AUD risk gene variants and the influence of polygenic risk background in a human neural system.

NIH Research Projects · FY 2025 · 2015-05

Abstract The hematopoietic system is one of the organ systems most vulnerable to radiation induced short- and long- term damage. Efficient recovery from pathological or medically induced bone marrow failure is dictated by the intrinsic sensitivity of the hematopoietic stem cell and the bone marrow environment niche. Identification of molecules that affect hematopoietic recovery is essential to the development of novel medical countermeasures against radiation damage. Our preliminary studies suggested that loss of even a single copy of Bccip confers hypersensitivity of mice to radiation-induced hematopoietic syndrome and lymphomagenesis, and the recruitment of BCCIP to DNA damage sites are dependent on PARP1. We hypothesize that Bccip haploinsufficiency can sensitize the hematopoietic stem cells to radiation killing, impair the long-term competency of stem cell to reconstitute the hematopoietic system, and/or affect the bone marrow niche’s capacity to nourish hematopoiesis. In Aim 1, a series of long-term and short-term experiments will be used to determine whether Bccip haploinsufficiency enhances the killing of hematopoietic stem and progenitor cells, impair stem cells’ capacity to reconstitute the bone marrow, and diminish the ability of bone marrow niche to nourish the hematopoiesis. We also hypothesize that Bccip haploinsufficiency alters the bone marrow progenitor cell susceptibility to tumor initiation and subsequent tumor progression. In Aim 2, we will test this hypothesis by examining the tumor clonality and defining the landscapes of chromosome rearrangements in the tumors formed in wild type and Bccip haplo-insufficient mice using newly developed genomic and computational approaches. In Aim 3, we will determine the PARylaiton dependent mechanism by which BCCIP is recruited and retained at the DNA damage sites. Completion of these studies will elucidate a unique role of Bccip in modulating hematopoiesis after radiation damage and in suppressing radiation-induced tumorigenesis.

NIH Research Projects · FY 2025 · 2012-09

Air pollution and tuberculosis (TB) represent two of the most devastating and life-threatening global public health problems. With hundreds of million persons living in urban slums worldwide, vulnerable, marginalized and hard- to-reach populations are most affected by indoor and outdoor air pollution and Mycobacterium tuberculosis (Mtb) transmission. This project explores if and how air pollution fine particulate matter (PM2.5) exposure increases the transmission of Mtb from TB index cases to their household contacts (HC) in a high TB incidence urban slum area (Namuwongo) in Kampala/Uganda. We have previously shown that air pollution PM2.5 exposures suppress essential, protective, Mtb-specific, human innate and adaptive host immune responses. A key unanswered question of great significance to global TB control efforts is whether air pollution exposure increases the risk for transmission of Mtb in vulnerable communities, both through increased infectiousness (of the TB index cases) and susceptibility to Mtb infection (of their contacts). We hypothesize that inhalational exposure to PM2.5 increases the infectiousness of TB index cases by increasing their source strength i.e. the Mtb content in their respiratory aerosols, and by triggering cough. We further hypothesize that PM2.5 exposure suppresses protective immune responses leading to increased susceptibility to Mtb infection in the HC of the TB index cases. Both would be expected to increase transmission of Mtb in the community. To address our hypotheses, we will evaluate (1) the source strength of TB index cases and new Mtb infection in their HC (Mtb transmission) (SA1), (2) personal air pollution (PM2.5) exposures (SA2) and (3) how personal air pollution (PM2.5) exposure is associated with PM load in airway macrophages, source strength, and Mtb transmission to HC (SA3). To assess the source strength of the TB index cases, we will quantify the Mtb load in their respiratory aerosols using a novel face mask sampling approach. We will also determine TB index case cough frequencies with the Leicester Cough Monitor, assess chest radiographs and time to culture positivity (MGIT culture). Transmission of Mtb from TB index cases to HC will be ascertained by tuberculin skin test and QFN-Plus blood test between weeks 0 and 8. All measures combined reflect infectiousness of TB index cases. Exposure to PM2.5 of TB index cases and a random sample of HC will be studied using multiple approaches: `gold standard' gravimetric (UPAS) and real- time air monitoring between weeks 0 and 8, assessment of PM load in airway macrophages obtained by sputum induction of TB index cases and in randomly selected as well as newly Mtb-infected HC, stationary household PM2.5 and outdoor PM2.5 monitoring, and household study with questionnaires. Associations between TB index case source strength, Mtb transmission and PM2.5 exposure will be sought using linear modeling, to identify both main effects and modifying factors. Deciphering environmental factors that contribute to transmission of Mtb in vulnerable urban populations will provide much needed data for mandated public health actions and improved TB control approaches worldwide.

NIH Research Projects · FY 2025 · 2012-08

Project Summary/Abstract Tumors have distinctive metabolism. These distinguishing metabolic features enable both cancer diagnosis and therapy. Due to fast glucose metabolism, tumors light up by fluorodeoxyglucose PET imaging. Due to enhanced nucleotide synthesis, they are susceptible to antifolates, which are first-line therapies for leukemia (methotrexate) and lung cancer (pemetrexed). New facets of tumor metabolism continue to be discovered. Previously, co-PIs White and Rabinowitz contributed to the identification of autophagy and macropinocytosis as non-canonical means of cancer cell nutrient acquisition. Recently, they identified thrifty energy metabolism in tumors: by shedding normal tissue functions, tumors can grow and divide despite making less energy than most normal tissues. While valuable, up to now studies measuring tumor metabolic activity (e.g. with isotope tracing and mass spectrometry) have mainly studied the tumor as a whole. In the current era of immunotherapy, the insufficiency of this approach is painfully clear. Whole tumor measurements confound the metabolism of cancer cells and cancer-fighting immune cells. To resolve metabolism in tumors with cell type specificity, new methods are needed. Here we will develop and deploy such methods. The resulting knowledge will provide the basis for manipulating metabolism more effectively to help fight cancer. Aim 1 will determine, in state-of-the-art mouse modes, in vivo central metabolic fluxes (glycolysis, TCA) across different tumor cell types and during active response to immunotherapy. Aim 2 will measure how different tumor cell types obtain nucleotides. Nucleic acid synthesis is a clinically proven cancer vulnerability but at the same time, nucleotide synthesis inhibitors can also impair immune cells. We will assess nucleotide synthesis and scavenging, and how it changes in response to anti-folates. Defining the dynamics of nucleotide metabolism in vivo with cellular resolution will inform the metabolic relationship between tumor and immune cells in the tumor microenvironment. Aim 3 will explore the interplay between autophagy, metabolism, and antitumor immunity. Autophagy supports tumor cell metabolism, and host autophagy supplies nutrients to tumors and dampens antitumor immunity. We will use mouse genetics, nutrient supplementation, single cell transcriptomics, and in vivo isotope-tracing of host and tumor metabolism to define underlying mechanisms. The net impact of these efforts will be (i) broadly useful methods for tracking metabolism in action across cell types, (ii) fundamental knowledge of the metabolic feedstocks and pathways used by different tumor cells, and (iii) next-generation metabolic strategies for treating cancer.

NIH Research Projects · FY 2026 · 2012-04

Project Summary The focus of this research program, which has been NIH-funded since 2012, is the influence of paternal psychostimulant self-administration on the physiology and behavior of subsequent generations (i.e. offspring and grand-offspring) using rat models. Our work previously demonstrated that sire cocaine self-administration reprogramed the germline epigenome resulting in decreased cocaine reinforcing efficacy in the male progeny. The current application expands our focus on cocaine alone to encompass the transgenerational effects of paternal methamphetamine (meth), which will be compared and contrasted with cocaine. The proposed research will examine the mechanisms whereby information is passed to meth-sired offspring (and potentially grand-offspring) through epigenetic changes in sperm. We also will define epigenetic and transcriptional profiles in the nucleus accumbens of cocaine- and meth-sired offspring that may underlie the respective influences on psychostimulant self-administration. Specific Aim 1 will examine the behavioral consequences of paternal meth self-administration on psychostimulant self-administration in male and female offspring (F1) and grand-offspring (F2). Intriguingly, our preliminary results indicate that meth and cocaine produce opposite effects on psychostimulant reinforcing efficacy in male offspring. In contrast to our prior results with cocaine, preliminary data indicate that paternal meth self-administration results in increased self-administration of, and motivation for, this psychostimulant selectively in male offspring. Drug naïve F1 rats will be used to generate an F2 generation, where the acquisition, maintenance and reinforcing efficacy of meth will be assessed just as in F1 offspring. In Specific Aim 2 we will assess epigenetic changes in sperm through which paternal psychostimulant self-administration may influence the behavior of offspring. Potential transgenerational cocaine- and meth-induced epigenetic alterations (small noncoding RNAs and DNA methylation) in sperm will be evaluated. Finally, genome-wide assessments of psychostimulant transgenerational effects have not yet been performed on neurons in the nucleus accumbens, a brain region that plays a critical role in modulating psychostimulant-induced behaviors. The experiments in Specific Aim 3 are designed to interrogate the landscape of accessible chromatin and gene expression by coupling single-nuclei ATAC-seq and single-nuclei RNA-seq analyses in the accumbens of experimentally naive meth-sired, cocaine-sired and saline-sired rats. Collectively, the experiments described in this application will use state-of-the-art cellular, molecular and behavioral methodologies to examine epigenetic mechanisms whereby psychostimulant-associated information can be transmitted from sires to offspring and grand-offspring. These cross-generational studies represent a novel strategy to identify transcripts related to risk or protective factors for psychostimulant self- administration, which will illuminate new targets for therapeutic development.

NIH Research Projects · FY 2026 · 2011-08

PROJECT SUMMARY This is a competitive renewal application for continued support of the Rutgers Summer Undergraduate Research Fellowship (SURF) program. The SURF program provides undergraduates with 10-week basic science and translational hands-on research experiences in environmental health sciences, along with weekly networking and career activities. In this renewal application, funds are requested to support fellowships for at least 10 students. Students are admitted to the SURF program after a review of scholastic achievement, letters of rec- ommendation, and personal statements. Recruitment strategies are in place to continue to attract students from across the country and ensure broad participation from various scientific backgrounds. Under the mentorship of NIH-funded Rutgers faculty, SURF students conduct hands-on research projects focused on environmental health sciences and participate in an innovative team-based field study that involves sampling and analysis of traditional and emerging contaminants, including toxic metals, micronanoplastics, or particulate matter from wild- fire smoke. Students meet with mentors weekly to receive one-on-one training, ensure the continued develop- ment of research skills, and evaluate the progress of scientific projects. The SURF program draws upon the resources, infrastructure, and expertise of faculty who are members of the Environmental and Occupational Health Sciences Institute, and part of our NIEHS T32 Toxicology predoctoral and postdoctoral training grant and NIEHS P30 Center for Environmental Exposures and Disease at Rutgers University Ernest Mario School of Pharmacy, Robert Wood Johnson Medical School, and School of Public Health. Students also participate in weekly seminars and workshops on topics of career development, responsible conduct of research training, laboratory safety training, and environmental research. Working with SURF leadership, each student develops a customized training experience that leverages the core training and research activities but provides tailored experiential learning based on their competency gaps and interests. The summer program culminates in the preparation of oral presentations and abstracts (written and graphical) that describe key research findings. The SURF Program Directors have an extensive evaluation plan to ensure the continued development of the fellow- ship program and dissemination of innovative training approaches and successful metrics. During the prior fund- ing period, R25-funded trainees have had outstanding outcomes, including numerous national awards, co-au- thorship on peer-reviewed publications, and matriculation into professional and graduate programs in toxicology. Renewal of this R25 application will provide an opportunity for Rutgers faculty to continue to train the next gen- eration of toxicologists and environmental health scientists.

NIH Research Projects · FY 2025 · 2011-04

ABSTRACT Transient Receptor Vanilloid 6 (TRPV6) and its close relative TRPV5 are Ca2+ selective epithelial ion channels. The membrane phospholipid phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is the endogenous ligand of these channels that is required for their activity. These channels also undergo Ca2+-induced inactivation which is mediated by binding of Ca2+-calmodulin (CaM) to the channel. TRPV6 and TRPV5 are constitutively active, and their level of activity is determined by the balance between the activating PI(4,5)P2 and the inhibitory CaM. In the previous funding period, we identified the PI(4,5)P2 binding site in TRPV6 using homology modeling and site directed mutagenesis, which was essentially identical to the experimentally determined PI(4,5)P2 binding site in the closely related TRPV5. CryoEM structures for CaM and TRPV6 or TRPV5 also became available, showing a highly consistent picture of one CaM molecule binding to the channel tetramer, and blocking the pore in a fashion consistent with our experimental results. An important development during the previous funding period was the finding that loss of function mutations in TRPV6 are associated with chronic pancreatitis. The likely mechanism is reduced Ca2+ removal from pancreatic fluid in the acini or the ducts leading to increased Ca2+ and premature activation of pancreatic digestive enzymes. Given the crucial role of Ca2+ in digestive enzyme activation, small molecules that increase the activity of TRPV6 can, in principle, be used as novel therapeutic approach to treat pancreatitis. The overall goal of this application is to gain molecular insight into how the two key endogenous regulators PI(4,5)P2 and CaM gate TRPV6 and to use our molecular level understanding of their regulation to identify small molecules that increase their activity. We will use a combination of computational and experimental approaches to decipher the molecular mechanism of PI(4,5)P2 activation of TRPV6 and TRPV5 in Aim1, to determine the relationship between CaM and PI(4,5)P2 regulation of TRPV6 in Aim 2, and to identify small molecules that increase TRPV6 and/or TRPV5 activity by interfering with CaM inhibition in Aim 3. This work will further our understanding of TRPV6 and TRPV5 gating, and small molecules that enhance the activity of TRPV6 or TRPV5 can be used in future experiments to explore the possibility of using this approach to treat or prevent pancreatitis and kidney stones.

NIH Research Projects · FY 2025 · 2010-08

SUMMARY/ABSTRACT This application seeks to renew the IRACDA at Rutgers - INSPIRE Postdoctoral Training Program- that began at Robert Wood Johnson Medical School (RWJMS) in August of 2010. RWJMS is now part of Rutgers University, expanding the pool of postdoctoral fellows and laboratories. INSPIRE would like to continue working towards its two goals: (1) to train the next generation of scientist-educators by creating an innovative postdoctoral training model and (2) to encourage undergraduates underrepresented in research to consider careers in biomedical sciences. The success of INSPIRE is evident in that (1) 74% of INSPIRE Fellows have obtained academic positions after research training at Rutgers laboratories and mentored teaching at our partner institutions, Medgar Evers College – CUNY, New Jersey City University, NJ and William Paterson University, NJ, three institutions that educate students underrepresented in biomedical research; and (2) INSPIRE has promoted improved and new science courses, Scientific Teaching training by partner school faculty, and increased research experiences for the undergraduates at partner schools. To increase the impact of INSPIRE the program will add a 4th year of formal training for the Fellows, and will expand mentoring programs for the partner school students. The proposed INSPIRE IRACDA program will include 3 years of funded postdoctoral training program for 5 trainees per year, with a 4th year of training to be supported by the Rutgers Research Mentors. Some INSPIRE Fellows will be recruited to become Rutgers faculty, as Rutgers Presidential Postdoctoral Fellows, a program to build Rutgers faculty diversity. Trainees will participate in mentored research in Rutgers laboratories; a structured career development program; a structured educational component including intensive training in scientific teaching during year 1; and mentored teaching and course development at partner schools during years 2 and 3, and focused training in academic writing in year 4. The program will promote research and teaching interactions between partner school students and faculty with faculty and postdoctoral trainees at a research-intensive university. Program evaluation will measure (a) undergraduate student application and entry into research programs and awareness of research careers; (b) curriculum development at partner schools of 4 science courses to increase student engagement in STEM courses; (c) research collaborations between Rutgers and its partner schools facilitated by ongoing summer research by partner school undergraduates at Rutgers, supervised by the Fellows and (d) entry of 70% of INSPIRE postdocs, including underrepresented trainees, into education/research careers, including becoming Rutgers research faculty.

NIH Research Projects · FY 2024 · 2010-07

PROJECT SUMMARY Congenital heart disease (CHD) is a significant cause of neonatal mortality and morbidity worldwide. Defects in the aortic arch artery (AAA) and its major branches are among the most severe malformations that cause CHD. The AAA and its branches arise through the asymmetrical remodeling of the three bilaterally symmetrical pairs of the pharyngeal arch arteries (PAAs), 3, 4, and 6. Defects in the left 4th PAA are particularly devastating, leading to the interruption of the aortic arch type B (IAA-B). IAA-B disrupts the systemic circulation and is lethal after birth. The genetic causes of IAA-B are mostly unknown, underscoring a critical need to understand genes and mechanisms regulating the formation of the left 4th PAA. PAA formation is a multi-step process: PAA endothelial cell (EC) progenitors arise in the second heart field (SHF) by E7.5 and populate pharyngeal arches within the next two days, culminating in the formation of a plexus of small SHF- derived blood vessels between by E10.0 in the 4th arch. Following its formation, the EC plexus gradually remodels by merging in the middle of the 4th arch into one large vessel, the 4th PAA. The deletion of Fn1 or integrin a5b1 in the Isl1 lineages results in the defective formation of the right and left 4th PAA and leads to IAA-B. My lab has demonstrated that cell-ECM interactions regulated by Fn1 and integrin a5b1 function reiteratively, at multiple stages to regulate the 4th PAAs’ formation and remodeling in a cell-type-specific manner. In this grant, we present data implicating Fn1 and integrin a5b1 in maintaining the ipsilateral patterning of the SHF-derived vasculature in the pharynx at E9.5. Our data show that the deletion of Fn1 or integrin a5b1 in the Isl1 lineage disrupts symmetrical allocation of SHF-derived cells to the left and right pharyngeal arches, causing an imbalance in the number of SHF-derived cells on the left vs. the right. At a later stage, between E10 and E10.75, Fn1 and integrin a5b1 regulate vascular patterning by mediating the remodeling of the pharyngeal EC plexus into the 4th PAA. Our published data show that Fn1 and integrin a5b1 mediate the plexus-to-PAA remodeling in an EC-non-cell- autonomous manner by regulating the expression of negative vascular guidance cues and the activation of VEGFR2 and Erk1/2 in the 4th arch. To determine the mechanisms by which Fn1 and integrin a5b1 regulate the patterning of SHF-derived vasculature, we will use live imaging, quantitative immunofluorescence microscopy, genetic epistasis analysis, and single-cell RNAseq to test two hypotheses 1) that Fn1 and integrin a5b1 expressed in the Isl1 lineage maintain the balanced allocation of SHF-derived cells in the pharynx by regulating ECM assembly and ipsilateral SHF cell migration and 2) that Fn1 and integrin a5b1 regulate plexus-to-PAA remodeling by modulating the expression of EC guidance cues and the activation of Erk1/2. Completing the proposed studies will provide essential insights into the functions of cell-ECM interactions in the development of SHF-derived vasculature and CHD pathogenesis.

NIH Research Projects · FY 2024 · 2009-07

Project Summary/Abstract PALB2 is tumor suppressor protein that physically and functionally links BRCA1 and BRCA2, the two major breast cancer suppressors, in the DNA damage response and tumor suppression. The 3 proteins function in a BRCA1-PALB2-BRCA2 axis to play essential roles in in homologous recombination (HR)-based DNA double strand break repair (DSBR) and cell cycle checkpoint control after DNA damage. These functions are critical for the maintenance of genome stability and suppressing tumorigenesis. In recent years, much has been learned about the mechanisms of PALB2 in HR-DSBR, but the mechanisms by which PALB2 functions with BRCA2 and BRCA1 to promote cell cycle checkpoint response remain elusive, and how normal PALB2 mutant cells survive and proliferate in vivo and evolve into cancer cells is poorly understood. In this proposal, we will deploy a combination of state-of-the-art in vitro and in vivo approaches to answer the above open questions. In Aim 1, we will use phosphoproteomics, targeted proteomics and functional studies to identify key factors and mechanisms of PALB2-mediated G2/M checkpoint control. In Aim 2, we will build upon our recent discovery of NFκB activation in Palb2 mutant mice and use a combination of systemic knockin, conditional knockout, lineage tracing and organoid models to define the mechanisms of PALB2-associated mammary tumorigenesis and the role of NFκB in the process. In Aim 3, we will determine the genetic mechanisms of our newly discovered complementary relationship of PALB2 and BRCA1 in mammary tumor development by testing the roles of RAD52 and RNF168 in the respective models.

NIH Research Projects · FY 2026 · 2008-01

Summary Oxidative stress affects the function of signaling molecules through posttranslational modifications of cysteine residues, which in turn regulate survival and death of cardiomyocytes (CMs) during myocardial stress. We have shown that thioredoxin1 (Trx1), a 12 kD oxidoreductase, modulates the function of signaling molecules, including HDAC4, AMPK, mTOR, and Atg7, in the heart by modulating their cysteine residues through either thiol-disulfide exchange or transnitrosylation. Here we will investigate a novel role of cysteine oxidation in mediating the degradation of sarcoplasmic reticulum (SR)/endoplasmic reticulum (ER) Ca2+ ATPase 2a (SERCA2a) during heart failure and its regulation through non-canonical interaction between p22phox, SERCA2a and Trx1. SERCA2a mediates Ca2+ reuptake into the SR/ER. Reduced contractility in the failing heart caused by impaired Ca2+ cycling between the sarcoplasm and SR and SERCA2a downregulation are important drivers of heart failure progression. Although rescue of SERCA2a using adeno-associated virus 9 (AAV9)-mediated delivery is effective in animals, it does not appear fully effective in heart failure patients. Complex posttranslational modifications of SERCA2a, including oxidation, acetylation and SUMOylation, may not allow effective restoration of SERCA2a function with the current strategy. Oxidation of SERCA2a induces proteolysis through the ubiquitin proteasome system. We discovered that mice with cardiac-specific deletion of p22phox are more susceptible to heart failure during pressure overload (PO) even though bulk production of reactive oxygen species is attenuated. In depth studies of the underlying mechanism suggest that endogenous p22phox binds to SERCA2a, protects SERCA2a from oxidation of cysteine residues, including Cys498, and prevents Smurf1/Hrd1-mediated proteasome degradation of SERCA2a during oxidative stress in vitro. These results suggest that p22phox plays a previously unrecognized role in the regulation of the function of SERCA2a function through direct interaction. We will test the following hypotheses: 1. p22phox protects the heart against heart failure in response to PO by stabilizing SERCA2a through direct interaction. 2. p22phox stabilizes SERCA2a against oxidative stress by mediating SERCA2a-Trx1 interaction, inhibiting oxidation of SERCA2a cysteines, including Cys498, and preventing SERCA2a-Smurf1/Hrd1 interaction. To test these hypotheses, we will use newly generated genetically altered mouse models, including p22phox cardiac specific knock-out and SERCA2a(C498S) knock-in mice. We will use proteomic approaches to evaluate posttranslational modification of cysteine residues in SERCA2a and demonstrate thiol-disulfide exchange between SERCA2a and Trx1. Our study will not only demonstrate a novel noncanonical function of p22phox but also clarify the detailed mechanism through which oxidation of SERCA2a at previously unrecognized cysteine residues leads to proteasomal degradation during PO. The study will provide useful information for efficient restoration of SERCA2a function during heart failure through modulation of cysteine oxidation.

NIH Research Projects · FY 2024 · 2006-09

Abstract This is a renewal application for the Rutgers University CounterACT Research Center of Excellence, leaders in the discovery and development of drugs to treat poisoning from exposure to chemical threat agents. The Center focuses on vesicants including sulfur mustard, nitrogen mustard, and other mechanistically related chemicals and is organized into three Research and Development Projects, each focused on a major vesicant target: the skin, the cornea and the lung. The overall goal of the Center is to identify specific targets in these tissues that can be used for therapeutic intervention. To accomplish this goal, mechanistic research using appropriate translational models for the skin, cornea and lung is being performed by scientific experts in each of these areas. The Research and Development Projects are supported by a Pharmacology and Drug Development Scientific Support Core and a Pharmaceutics and Medicinal Chemistry Scientific Support Core, each consisting of investigators with considerable expertise in drug discovery, delivery and development. Through mechanistic research, the Rutgers Center has 1) discovered and prioritized several efficacious lead compounds and 2) developed innovative formulations and delivery systems that augment pharmacological activity and improve pharmacokinetic profiles. Three lead compounds, two for mustard-induced ocular injury and another for mustard-induced lung injury, are FDA approved for other indications, and are being advanced for use as mustard countermeasures. Furthermore, IND-enabling studies have been completed on a lead compound discovered to be effective against mustard-induced skin injury. The Rutgers CounterACT Center has established strategic corporate partnerships for advanced drug development with lead compounds and support by BARDA. A Research Education Core directed at students, postdoctoral trainees, technical staff, and new and established investigators has been highly successful at training a skilled workforce in countermeasures. During the next grant period, we will 1) identify and develop second generation drug products with improved effectiveness, 2) disseminate key findings about mechanisms of toxicity and novel testing strategies and models, 3) advance current drug leads through the FDA approval process and 4) expand training and workforce development opportunities. By meeting key research, development, and training milestones, the Center is well-poised to advance new and repurposed drugs through the approval process for treating vesicant poisoning in humans.

NIH Research Projects · FY 2024 · 2005-09

Project Summary The goal of this competing continuation is to examine the antecedents and health consequences of alcohol misuse from adolescence through early midlife (the 30s). There is a paucity of longitudinal studies on alcohol misuse that extend through early midlife in the general population, and the existing knowledge base comes from studies of singletons, which are prone to confounding. We propose to address these limitations by bringing together two longitudinal studies of Finnish twins, FinnTwin12 (FT12) and FinnTwin16 (FT16), ns = 5178 and 5563, ~50% female. The innovative longitudinal twin design, which allows us to control for confounding factors through the comparison of exposures and outcomes within families and within individuals, will add much needed rigor to our understanding of the antecedents and consequences of alcohol misuse through early midlife. FT12 was recruited at age 12 and subsequently assessed at ages 14, 17, and 22. Under this renewal, we will conduct an early midlife (age 35) assessment of the FT12 twins (online surveys regarding their current health, behavior, and environments; a salivary DNA sample; and, for a subsample of participants who have been intensively studied since age 14, diagnostic psychiatric and life history calendar interviews and laboratory-based neurocognitive and health measures) and the twins’ spouses (online survey only). FT16 was recruited at age 16 and subsequently assessed at ages 17, 18, 25, and 35. Finland offers a unique environment for conducting the proposed research, including record linkage to comprehensive national registries and a modern biobanking infrastructure. Guided by a multilevel developmental contextual framework, our aims are to: (1) Characterize patterns of alcohol misuse from adolescence to early midlife. (2) Identify factors associated with trajectories of alcohol misuse between adolescence and early midlife. These include person-level factors such as polygenic predispositions, personality, and neurocognitive functioning; environmental factors such as parents, peers, spouses/partners, life events, parenthood, employment, and education; and internalizing, externalizing, and other substance use problems. (3) Examine the health outcomes associated with trajectories of alcohol misuse, including measures of physical health, sleep, and life satisfaction. Finally, we will examine the generalizability of effects using the U.S. nationally representative Add Health sample (n = 20,745), which has comparable measures and assessments, including a sibling component. The results will provide important information about alcohol misuse and its consequences through the understudied early midlife period. Through partnerships with collaborators in the arts and mass communications, we will translate and disseminate our findings to the public through nontraditional creative content to increase accessibility of scientific results. In sum, this project will contribute to NIAAA’s goals to identify factors associated with alcohol misuse across the lifespan; delineate the effects of alcohol misuse on health; and enhance the public health impact of research.

NIH Research Projects · FY 2026 · 1997-04

The Center for Environmental Exposures and Disease (CEED) at Rutgers University is a recognized leader in New Jersey (NJ) - the most densely populated state in the Union, with many of its residents living in close proximity to major congested roadways, and industrial and commercial centers. NJ is also the home of more Superfund sites than any other state due to a long history of poorly regulated industry, as well as active efforts to identify contamination. Over 85% of the state’s land area is already ‘built-out’ or preserved, meaning there will soon be no remaining undeveloped land. Thus, NJ is a microcosm that reflects the longstanding and emerging environmental problems confronting the entire nation. The strategic vision of CEED is to address the environmental health concerns of all NJ residents. We aim to accomplish our strategic vision by engaging community members, organizations, and agencies as advisors and partners through all stages of our research, providing critical investment and infrastructure for multidisciplinary collaborative research, and training the next generation of environmental health scientists. Our approach to achieve this vision is to identify and prioritize the most urgent and hazardous environmental health concerns through an engaged community advisory board and community-driven research initiatives that are integrated with the innovative translational research of CEED scientists. Over the next five years, CEED will confront environmental health-related disease by working with affected communities at the center of our efforts. Through bidirectional partnerships, rigorous science and training, cutting edge technologies, and effective communication, CEED will work to reduce health risks within NJ, and thereby improve public health across the state, the region, and the world.

NIH Research Projects · FY 2026 · 1997-03

Rutgers Cancer Institute is a matrix/consortium with Princeton University (PU) and is New Jersey’s only NCI-designated Comprehensive Cancer Center, serving a population of 9.2 million. The cancer institute focused the strengths and resources of the major research institutions in NJ on addressing the cancer burden in its catchment area (NJ). Rutgers Cancer Institute is an independent institute of Rutgers University (RU), optimizing its potential for growth and transdisciplinary collaboration. In FY2023, the cancer institute received over $140 million of institutional support (RU and PU, the State of NJ, the RWJBarnabas Health System [RWJBH], philanthropy). Metrics of success in this grant period include increases in: Center Members, PU Members, cancer focused peer reviewed support, NCI Institutional Training Grants, space, patient visits, interventional therapeutic trial accrual, State Support, and counties covered by ScreenNJ. Steven K. Libutti remains Cancer Center Director, Vice Chancellor for Cancer Programs for Rutgers Biomedical and Health Sciences, and RWJBH Senior Vice President for Oncology Services since 2017. Rutgers Cancer Institute entered into an innovative Integrated Practice Agreement with RWJBH that provides a financially sustainable model for provision of high-quality, cutting-edge, academic oncology services, and enabled system-wide restructuring of the clinical trial enterprise and a significant increase in accruals. Driven by its Strategic Plan, Rutgers Cancer Institute increased high impact center science; supported efforts to train the next generation of cancer researchers and leaders; and increased the impact of the Community Outreach and Engagement Office on its catchment area. The CCSG Research Programs all demonstrated enhanced collaborations and scientific impact: 1) Cancer Metabolism and Immunology, 2) Genomic Instability and Cancer Genetics, 3) Cancer Pharmacology, 4) Clinical Investigations and Precision Therapeutics, and 5) Cancer Prevention and Control. The Research Programs are supported by shared resources (Biomedical Informatics, Biostatistics, Biorepository and Histopathology, Genome Editing, Comprehensive Genomics, Immune Monitoring/Flow Cytometry, Metabolomics and Cancer Prevention/ Outcomes Data Support) and other services. Rutgers Cancer Institute’s future is guided by a Strategic Plan that includes increasing access for NJ residents to screening (new ScreenNJ mobile unit); increasing trial accruals through an expanded clinical research infrastructure; improving faculty and trainee mentorship and leadership; increasing multi- project grants through strategic investment; and operationalizing the Jack and Sheryl Morris Cancer Center, NJ’s only free-standing cancer hospital.

NIH Research Projects · FY 2026 · 1994-12

Parathyroid hormone (PTH) is an essential regulator of calcium homeostasis and became the prototypic osteoanabolic hormone for treating osteoporosis. On the other hand, hyperparathyroidism, with its consequent bone breakdown, is a major problem in and of itself and in chronic renal failure. The hormone acts through its G-protein-coupled receptor on the osteoblast to elicit enhanced bone resorption by the osteoclast. To do this, the osteoblast produces receptor activator of nuclear factor kappa-β ligand (RANKL) in response to PTH. RANKL is widely considered a principal mediator of PTH-induced bone catabolism and has been associated with bone loss in hyperparathyroidism. Our work in the last cycle of this grant revealed that PTH activates cAMP/protein kinase A (PKA) to inhibit salt-inducible kinases (SIKs) and requires protein phosphatases (PPs) to stimulate Rankl expression in osteoblasts. Knockdown of salt-inducible kinases (SIKs) 2 and 3 and cAMP- regulated transcription coactivators 2 and 3 (CRTC2 and 3) indicate that all four are part of this pathway. Moreover, inhibition of serine-threonine protein phosphatases decreased both PTH-induced Rankl expression and the stimulation by PTH(1-34) of CRTC2 and 3 translocation into the nucleus. Upon entry into the nucleus, these CRTCs associate with unknown basic leucine zipper domain (bZip) transcription factor(s), activating Rankl transcription through binding to its distal PTH-responsive cAMP-response elements (CREs). From these data of cells in culture and preliminary data in vivo, we have developed the central hypothesis that SIK/PP regulation of CRTC2/3 is essential to their function in the skeleton, and PTH controls this regulation. The long- term goals of this work are to delineate the signaling and transcriptional regulatory mechanisms conveying PTH action in bone. Consequently, the specific aims to test our hypothesis of this resubmitted competing renewal proposal focus on the SIK and PP regulation of CRTC2/3 function and action on Rankl, and will, 1) determine the mechanism of PTH regulation of Rankl transcription in osteoblasts by a. elucidating the regulatory mechanisms involved in CRTC2/3 nuclear translocation in osteoblasts, b. identifying the bZip transcription factor(s) responsible for CRTC2/3-induced Rankl transcription, 2) determine the role of CRTC2/3 in bone development and PTH’s anabolic and catabolic effects by a. determining the site and role of osteoblast CRTC2/3 in bone development, b. ascertaining the effects of deletion of CRTC2/3 on PTH’s anabolic and catabolic actions in bones of adult mice. The results of this work will make major contributions to our knowledge of how PTH exerts its nuclear effects on skeletal function through the SIK/PP/CRTC/bZip pathway. In so doing, the data will also provide new perspectives into treatment of disorders of calcium metabolism and other severe diseases functioning through the PTHR1 pathway such as Jansen’s metaphyseal chondrodysplasia and McCune-Albright syndrome.

NIH Research Projects · FY 2026 · 1992-09

Atlantic Center for Occupational Health and Safety Training Overall Abstract The Atlantic Center for Occupational Health and Safety Training has a long history of effective training, providing over 5.1 million contact hours of hazardous materials knowledge and skills, reaching over 506,000 workers since 1987. Our Center will train workers about safety issues during hazardous waste site clean-up, issues related to generation, treatment and storage of hazardous materials, emergency response, and disaster preparedness and response. Our Center will also provide career development training to unemployed and underemployed individuals, creating sustainable careers in environmental health and safety. The courses develop competency in workers to critically analyze dangerous situations, enable them to identify safe work practices, and make workplaces safer. Key principals of adult education are incorporated into our training. Courses are developed to include peer-learning, hands-on activities, and development critical thinking skills. The Center has a robust evaluation plan and will continue to expand the types of data collected on the impact of training on workplace practice, as well as how training can more effectively influence workplace safety culture. The Center adheres to the principles identified in the NIEHS Minimum Criteria Document. Training provided by the Center is aligned with several sections of the NIEHS Strategic Plan. The Center proposes to continue training through the HWWTP, HDPTP, and ECWTP programs. The HWWTP provides core hazardous waste and emergency response training. The training provides critical knowledge and skills for those requiring training under 29CFR1910.120 and other occupational safety and health standards. Our Center proposes to expand our capacity to reach underserved groups throughout Region 2 by adding The City College at the City University (CCNY) and The Work Environment Council of New Jersey (NJWEC). Over the next five years, the Center proposes 26,380 workers in 1,835 courses, providing 387,640 contact hours of training in the HWWTP. The HDPTP enhances the safety and health of current hazardous materials workers and chemical responders by delivering training to workers and volunteers responding to disasters. The courses focus on prevention and preparedness so that disaster response personnel are aware of safety and health hazards and mitigation techniques before they initiate a disaster response. The Center proposes to add the Northeast New York Coalition for Occupational Safety and Health (NENYCOSH) to provide training in the NY State Capital (Albany) Region. Over the next five years, the Center proposes to train 9,335 workers in 560 courses resulting in 76,415 contact hours in the HDPTP. The ECWTP provides training to unemployed and underemployed individuals. The program creates career paths in environmental health, allowing trainees to achieve sustainable employment. The program targets minority communities with high unemployment and high poverty rates in low-income areas in New York City and New Jersey. The Center proposes to add Make the Road New York to the ECWTP to increase the employment opportunities for disadvantaged immigrants. Over the next five years, the Center proposes to train 825 individuals in 50 cohorts, in 210 courses, providing 161,250 contact hours in the ECWTP.

NIH Research Projects · FY 2025 · 1989-06

ABSTRACT Ozone is a ubiquitous air pollutant and the main component of photochemical smog. It remains a federally regulated air pollutant of ongoing public health concern. Inhaled ozone irritates and damages the lung in both healthy and susceptible individuals, including children and the elderly. Ozone causes inflammation and constriction of the airways, reducing pulmonary function. Ozone also exacerbates asthma and chronic lung disease. Thus, elucidating mechanisms of toxicity is highly relevant in terms of identifying new strategies to reduce lung injury from ozone and potentially other air pollutants. Our studies are focused on macrophages, which we have demonstrated play a key role in both initiating and resolving inflammatory responses to ozone- induced tissue injury. These activities are mediated by distinct subsets broadly classified as proinflammatory M1 and proresolution M2 macrophages. Effective resolution of inflammation following tissue injury depends on metabolic reprogramming of macrophages from an M1 phenotype to an M2 phenotype, which involves a switch from glycolysis to oxidative phosphorylation as a source of energy. We discovered that this reprogramming is suppressed following ozone exposure. The goal of our studies is to analyze mechanisms underlying suppression of macrophage reprogramming. In recent studies we identified farnesoid-X receptor (FXR), a nuclear receptor important in regulating lipid metabolism, with anti-inflammatory activity, as central to promoting M1 to M2 macrophage reprogramming in the lung. Following ozone exposure, macrophage FXR activity is downregulated. This is associated with increased activity of proinflammatory M1 macrophages and reduced activity of proresolving M2 macrophages. We also found that microRNAs that regulate the proinflammatory transcription factor NFκB are dysregulated in macrophages after ozone exposure. As a consequence, there is protracted activation of NFκB signaling resulting in increased production of inflammatory mediators. We hypothesize that these mediators suppress FXR activity which prevents activation of the PPARγ coactivator (PGC-1β), an inducer of macrophage M1 to M2 metabolic reprogramming. To test this hypothesis, we will perform parallel studies in mice and humans and (1) Determine if persistent inflammation following ozone exposure and lung injury is due to impaired development of proresolution M2 macrophages, and assess whether this is caused by protracted activation of NFκB; (2) Analyze the role of FXR and its target, PGC-1β in the development of proresolution M2 macrophages in the lung following ozone exposure; and (3) Assess whether protracted activation of NFκB is a consequence of ozone-induced alterations in microRNAs regulating NFκB. Results of these studies will provide new mechanistic insights into ozone toxicity. This will have significant translational implications for the development of new strategies for preventing and treating the toxicity of ozone, and possibly other agents that induce inflammatory lung injury.

NIH Research Projects · FY 2026 · 1988-11

Project Abstract HIV remains a key health challenge, with fundamental aspects of the viral replication machinery remaining poorly understood. Building on our extensive studies of HIV-1 reverse transcriptase (RT) structure and function, we propose to further investigate RT’s roles within the reverse transcription complex (RTC) and the viral maturation process. Our previous work has provided fundamental insights into HIV-1 RT's molecular architecture, including key structures of RT complexed with diverse nucleic acid substrates and nucleoside, non-nucleoside, and other inhibitor types, and has contributed to elucidating molecular mechanisms of inhibition and resistance. We propose to address three critical knowledge gaps: (1) further definition of the specific conformational states and transitions during first- and second-strand DNA synthesis initiation and elongation; (2) the molecular interaction between RT and its partner host protein eEF1A and its functional role within the RTC; and (3) the role of the RT portion of Gag-Pol in HIV virion assembly and maturation. Using cutting-edge cryo-EM and X-ray crystallography, complemented by biochemical and virological studies through established and new collaborations, we will determine structures of RT complexes that capture these dynamic processes. Our preliminary data include novel structures of RT/DNA:RNA polypurine tract complexes, successful engineering of polyprotein constructs enabling the visualization of the RT and protease (PR) dimeric regions within Gag-Pol and Pol polyprotein structures by cryo-EM, and optimization of protein expression and structure determination for studying the RT-eEF1A interaction. Anticipated outcomes and insights include high- resolution structures detailing: 1) reverse transcription initiation phases and transition to elongation; 2) RT's interactions with eEF1A and rationale for its role in promoting reverse transcription; 3) how RT participates in virion assembly within Pol and Gag-Pol polyprotein intermediates; and 4) how targeted activators of cell kill (TACK) non-nucleoside RT inhibitors enhance PR dimerization and viral maturation, triggering HIV-specific cell death, with potential application to HIV cure strategies. These efforts also involve improvement of current processing strategies for recovering multiple conformations in cryo-EM data, given the inherent conformational heterogeneity of viral polyproteins. These studies will significantly advance our understanding of HIV replication and guide the development of novel therapeutic strategies, particularly through the identification of new druggable interfaces and further characterization of established (non-nucleoside inhibitors) and innovative inhibitor classes, such as TACK inhibitor molecules. Impact: This research will provide fundamental insights into HIV biology while opening new avenues for therapeutic intervention in the continuing fight against HIV/AIDS.

NIH Research Projects · FY 2025 · 1987-09

The Joint Graduate Program in Toxicology (JGPT) is an interdepartmental training program in mechanistic toxicology at Rutgers University. Founded in 1981, the JGPT has trained more than 200 doctoral students, postdoctoral fellows, and clinician scientists. Graduates of the JGPT have established distinguished careers in academia, industry, and government. The NIEHS T32 training grant is the driving force of the JGPT. This competitive renewal application requests funding for years 36-40 to support 8 predoctoral students and 3 postdoctoral fellows each year. The central mission of the JGPT is to provide talented and motivated predoctoral and postdoctoral trainees with rigorous didactic and laboratory training in contemporary mechanistic toxicology and in-depth expertise in their individual field of research. The program is highly interdisciplinary with students performing research rotations under scientists from varied scientific disciplines. Training is supported by an exceptional research environment centered at the Environmental and Occupational Health Sciences Institute (EOHSI) that includes state-of-the-art instrumentation and collaboration that synergizes with our NIEHS P30 Center of Excellence in Environmental Health Sciences. Rutgers University has designated “Environmental Health Sciences” a Signature Program. This affords our program strong institutional support with significant investment in research, training, and new faculty recruitment. JGPT trainees benefit from an outstanding seminar series and strong participation by leading scientists from the local pharmaceutical, chemical, and personal care products industry. Intensive efforts are expended to attract students and fellows of exceptional quality with prior research experience from broad scientific disciplines. Trainee accomplishments include high research productivity, strong publication and fellowship records, job placement, and honors and awards from professional organizations. The JGPT adapts to advances in the field of toxicology using comprehensive outcomes assessment. RELEVANCE: The unifying goal of the JGPT and this training grant is to prepare trainees to excel in the competitive and rapidly evolving arena of environmental health sciences. Toxicology is a core discipline in understanding the impact of chemicals on human health. For the last 35 years, this training grant has enabled Rutgers to educate scholars who have become leaders in academic, industrial, and governmental toxicology.