Rutgers Biomedical And Health Sciences

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Genetically engineered immune cells are an exciting and promising frontier for treating a wide range of complex diseases. However, hyperinflammatory toxicities such as cytokine release syndrome plague clinical trials, stymieing their widespread clinical adoption. Macrophages are innate immune cells that fulfill many roles in tissue repair, regeneration, and homeostasis and are important regulators of inflammation. However, they are significantly under-utilized as engineered immune cell therapies because significant knowledge gaps exist in understanding how to engineer synthetic gene circuits that work robustly in human macrophages. Tools do not yet exist for determining how macrophages should be biologically manipulated to activate desired effector functions (biological design principles). Tools also do not exist for determining what gene circuit architectures are needed to robustly induce desired gene circuit behaviors (gene circuit design principles). The overall goal for this proposal is to create a human macrophage design toolkit for engineering therapeutic macrophages. Our published and preliminary data demonstrate that we have developed tools that enable us to discover cell signaling interventions that can control macrophage effector functions (biological design principles) and gene circuit architectures that can exert robust behaviors in human macrophages (gene circuit design principles). Here we will apply both these approaches to elucidate biological and gene circuit design principles that can be used to engineer therapeutic macrophages that can suppress inflammatory cytokine secretion or induce anti- inflammatory cytokine secretion in inflamed tissues. We will elucidate biological design principles using an interpretable machine learning approach that we previous developed. This approach combines biochemical screening with predictive network modeling and machine learning to discover network mechanisms causally regulating cell phenotypes. We will elucidate gene circuit design principles using a recently developed ultra-high- throughput genetic screening approach (CLASSIC). This approach synthesizes and screens large, barcoded gene circuit libraries to associate gene circuit architectures with gene circuit behaviors. With these design principles we will engineer gene circuits for controlling IL-1β or IL-10 secretion in inflamed tissue contexts and validate these synthetic gene circuits in human monocyte-derived macrophages and THP-1 cells. In its entirety, this Trailblazer R21 project is a first step towards addressing the unmet need for design principles for engineering therapeutic macrophages. We envision that insights gained by this project will help establish engineered macrophages as a platform technology for treating a wide range of complex human diseases.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Lung cancer is the leading cause of cancer death among US men and women and accounted for more than 125,000 deaths in 2023—more than 20% of all cancer deaths. An important strategy to reduce mortality is lung cancer screening by low-dose computed tomography (LDCT), which has been estimated to reduce lung cancer mortality by up to 20%. In 2013, the US Preventive Services Task Force (USPSTF) began recommending LDCT lung cancer screening for people aged 55–80 years with a 30 pack-year history of smoking who currently smoke or had quit within 15 years; in 2021, these criteria were revised to include adults aged 50–80 with a 20 pack- year smoking history. Although uptake has been slow, there have been favorable shifts in diagnoses towards earlier stages. Still, nearly half of lung cancers today are diagnosed at distant stages, and inequities in lung cancer screening and mortality persist for several groups such as African Americans, women, and people with lower income, many of whom smoke menthol vs non-menthol cigarettes and at increasing rates. Menthol in cigarettes is associated with increased smoking initiation, propensity to progress to regular smoking, increased nicotine dependence, and decreased cessation success. Paradoxically, people who smoke menthol cigarettes have comparable levels of nicotine and carcinogen exposure biomarkers yet consume fewer cigarettes per day than people who smoke non-menthol cigarettes. Prior studies have concluded that menthol cigarette type need not be considered when estimating risk for lung cancer. However, these studies predated growth in the menthol cigarette market and the introduction of LDCT lung cancer screening, for which eligibility is determined largely by cigarettes smoked per day. This raises new questions about the potential indirect impact of menthol cigarette smoking on lung cancer screening and outcomes: if people who smoke menthol consume fewer cigarettes per day, are they less likely to meet screening eligibility criteria, despite having the same exposure to carcinogens? If true, people who smoke menthol face greater risk for later-stage diagnosis, and subsequently, mortality. This application will utilize the Tobacco Use Supplement to the Current Population Survey (TUS-CPS) and the Tobacco Longitudinal Mortality Study (TLMS) to evaluate the influence of menthol in cigarettes on lung cancer screening eligibility and outcomes. Under Aim 1, we will compare lung cancer screening eligibility for people who smoke menthol vs nonmenthol cigarettes, hypothesizing that menthol smoking will be associated with decreased probability of meeting screening eligibility criteria. We will also examine changes over time and differences by race/ethnicity, sex, and income through stratified analyses. Under Aim 2 we will evaluate lung cancer screening eligibility and menthol smoking exposure among individuals who have died of lung cancer using a case-case approach. We hypothesize that adjusted odds of menthol smoking will be higher for ineligible vs eligible cases and will evaluate heterogeneity by time (< 2013 vs after), race/ethnicity, sex, and income. We will also investigate the reasons for ineligibility among those not eligible (e.g., pack-years vs age) and if this differs by menthol status.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Cadmium, a non-essential metal, is persistent in the environment and highly detected in pregnant people. Non-smokers are primarily exposed to cadmium via the intake of contaminated foods (e.g., cereals, breads, vegetables). During pregnancy, cadmium concentrates in the placenta and can interfere with biological functions and processes including hormone production and nutrient transfer. As a result, maternal exposure to cadmium may contribute to preterm birth, reduced birth weight, and impaired child growth. Placental transporters such as the breast cancer resistance protein (BCRP/ABCG2) may offer the fetus some protection by removing cadmium from the placenta and returning it to maternal circulation. In vivo and in vitro models show that BCRP reduces placental cadmium concentrations and cellular toxicity. In humans, a nonsynonymous BCRP polymorphism (C421A/Q141K) that is present in 17% of people is associated with ~50% lower placental BCRP protein levels. This F31 fellowship will first leverage existing data from an estimated 4,763 mother-child pairs across 13 cohorts in the NIH’s Environmental Influences on Child Health Outcomes (ECHO) program to determine whether a common variant in the BCRP gene may leave some fetuses more vulnerable to the adverse impacts of cadmium exposure. Second, we will utilize novel placental proteomics data on transporters from a single ECHO cohort (UPSIDE) to examine relationships between urinary and placental cadmium exposures and placental transport protein concentrations. Our aims are: (1) To examine prenatal cadmium concentrations in relation to gestational age at delivery and growth in the ECHO-wide cohort, considering effect modification by BCRP genotype. We hypothesize that higher prenatal cadmium will be associated with increased risk of preterm birth, earlier gestational age at delivery, smaller size at birth, and reduction-pattern growth trajectories. Further, we hypothesize that these associations will be stronger in pregnancies with the reduced-function BCRP polymorphism compared to the wild-type; (2) To examine placental BCRP protein concentrations in relation to prenatal cadmium concentrations in the UPSIDE cohort. We will explore associations with timing of birth and child growth. We hypothesize that placental BCRP protein concentrations will be inversely associated with prenatal cadmium concentrations. In addition to BCRP, we will consider protein concentrations associated with other less prominent gene variants and transporters involved in cadmium transport including a second BCRP variant and the Multidrug Resistance 1 [MDR1] transporter. This work will provide insight on the role of placental transporters in protecting against prenatal cadmium exposure and potentially identify a subpopulation of children at heightened risk of cadmium toxicity due to the BCRP variant. In addition, this research can be extended to improve our understanding of the developmental impacts of additional high priority contaminants that bind to BCRP including bisphenol A, perfluorooctanoic acid (PFOA), and zearalenone.

NIH Research Projects · FY 2025 · 2025-08

Abstract This T32 application seeks support for four postdoctoral lines at the newly established Rutgers Addiction Research Center (RARC), the largest comprehensive addiction research center in the U.S, to provide training in the etiology and consequences of alcohol involvement across the lifespan. The interdisciplinary training program will be led by a core team of 11 RARC-affiliated faculty mentors across a number of academic departments and centers at Rutgers who have active NIH funding portfolios and successful mentoring histories. Additional expertise will be provided by a group of 9 supporting faculty from RARC who are domain experts in related topics and/or methodological/ statistical experts. Faculty mentors contribute expertise in course of alcohol use across the lifespan, the underlying biological mechanisms (e.g., stress hormones, genetics, neurobiological) and psychosocial mechanisms (e.g., psychological, inter-personal, environmental), and ensuing consequences. Specific faculty expertise in related conditions include trauma/PTSD, mental and physical health, behavioral addictions including gambling and gaming, co-use of multiple substances, stress, sleep, and women’s health. Trainees will be able to customize advanced methodologic training specific to their research training goals across the areas of genomic analyses, longitudinal and intensive longitudinal analysis, digital health, community-based research, treatment implementation, and intervention design. An Internal Advisory Committee of five leaders at Rutgers and an External Advisory Board of four world-class researchers with substantial knowledge in alcohol research training and experience fostering the integration of diversity scholarship and alcohol research will monitor progress and provide guidance and recommendations. The training program will consist of a two-year sequence of mentored and didactic experiences and individually tailored professional development activities. Training includes hands-on mentored research with a primary mentor supported by a mentoring team as well as engagement in a faculty-led core seminar series, a grant writing sequence culminating in submission of an independent grant, journal clubs, and professional development panels, as well as trainee research talks and external speakers at an in-person retreat. First-year trainees will receive specialized training in state-of-the-art methodology, data analysis, and data science; diversity, equity, inclusion, and accessibility; and responsible conduct of research including practices to enhance research rigor and transparency. The program will host communication skills, mentorship training, and career development events and will support a strong scientific community through work-in-progress and shared interest groups and near-peer mentors. Finally, training faculty will receive mentorship training. Evaluation of individual trainees, faculty mentors, and the training program will be based on formal benchmarks. This training grant will launch the scientific careers of the next generation of alcohol researchers through interdisciplinary alcohol training in an untapped, rigorous, and highly resourced addiction research environment.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Cryptococcus (Cn) species have a global impact on human health as an environmental, opportunistic fungal pathogen that most commonly presents as pulmonary disease or meningoencephalitis (CM) causing an estimated 180,000 deaths annually in a range of patient populations. Despite knowing that T-cells are required for immune protection, little is known about the molecular mechanisms driving susceptibility to cryptococcosis. Our long-term goal is to identify genetic and immunological defects leading to susceptibility to Cn to inform pathobiology and personalized care. To address this goal, whole exome sequencing was performed on a cohort of previously healthy patients without known immunosuppression to identifying rare alleles associated with cryptococcosis and predicted to have deleterious functional consequences. For the first time, gene enrichment pathway analysis identified MTOR signaling deficits as enriched in this population. The goal of this proposal is to determine whether identified MTOR variants cause dysfunction in MTOR signaling associated with increased susceptibility to cryptococcal disease. Our central hypothesis is that MTOR deficiency predisposes patients to cryptococcosis and suggests functional MTOR signaling is required to clear Cn. This will be accomplished by: (Aim 1) Characterizing the immunologic consequence of MTOR deficiency on susceptibility to Cn using established murine models of cryptococcosis; and (Aim 2) Determining how patient MTOR variants alter cell signaling and function in patient lymphocytes. The results from this proposal will begin to elucidate for the first time mechanistically how human genetic variants in a single gene (MTOR) are associated with specific susceptibility to cryptococcosis in previously healthy individuals with no known immunologic defects. Thus, these results will provide novel insights into human disease susceptibility to Cn and reveal the protective immune responses affected by MTOR deficiency. Ultimately, expanding our definition for who is at risk for cryptococcosis and suggesting improved treatment opportunities for cryptococcosis patients identified to have aberrant MTOR signaling through pharmacologic modulation of cell-specific immunity. The applicants career goal is to become an independent scientist in the field of fungal immunology with a focus on understanding the mechanism by which specific host genetic defects lead to susceptibility of previously healthy patients to cryptococcosis. To meet this goal, the applicant will begin the process of transition to an independent scientist by beginning to apply for tenure-track faculty positions in fall 2024 cycle. Further, the applicant proposes a career development plan that will allow her to gain more experience in host genetic susceptibility/fungal immunology, grant writing, and leadership skills through practical experience, formal course work, and mentoring. Highly accomplished collaborators specialized in similar areas as the proposed research project will mentor the applicants career development and provide expertise on different aspects of the project.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Adolescent siblings of children with cancer are at-risk for psychological distress and report unmet needs in response to their sibling’s cancer diagnosis and treatment. However, most siblings do not have access to support services due to multiple barriers, including their absence from the hospital setting, inadequate staffing, billing challenges, and a paucity of widely disseminated evidence-based sibling interventions, which ultimately leads to significant service gaps and siblings being overlooked. Digital single-session interventions (SSI) significantly decrease psychological distress for adolescents in the general population; leveraging these high reach SSIs may overcome existing barriers to sibling care in psycho-oncology. Using a community-partnered approach, the goal of this proposal is to adapt, refine, and pilot test a Sibling SSI that aims to reduce psychological distress in adolescent siblings of children with cancer. Further, we aim to understand potential barriers and facilitators to implementation of this Sibling SSI. In Aim 1, the PI and the mentorship team will adapt an existing SSI to meet the specific needs of adolescent siblings of children with cancer. Adaptations will retain core SSI design principles and use solution-focused brief therapy to adapt an interactive, self-guided SSI for adolescent siblings. Adolescent siblings will participate in the adaptations through Think Aloud sessions (n=12). Aim 2 will evaluate the feasibility, acceptability, and preliminary outcomes of the Sibling SSI versus a waitlist control to decrease psychological distress in siblings (n=50). The team will assess key components of participant engagement to inform the future, fully powered clinical trial, including recruitment rate, retention, and preliminary outcomes. In Aim 3, semi-structured interviews will be conducted with key informants (i.e., adolescent siblings and parents of children with cancer, hospital administrators and psycho-oncologists, and administrators at community organizations) to identify potential barriers and facilitators to implementation using the RE-AIM framework. Findings will inform the development of specific implementation strategies and an implementation plan to be tested in future work. The long-term objective of this work is to support the families of children with cancer through effective, accessible interventions to improve their psychological well-being, with a specific emphasis on siblings. This K08 will provide opportunities for the PI to acquire skills and knowledge in: (1) adaptation of digital interventions with an emphasis on SSI and user-informed design, (2) the conduct of clinical trials focused on behavior change interventions, (3) implementation science for digital interventions, and (4) building and sustaining community partnerships for community-partnered research. This research and career development plan, which is supported by a multidisciplinary team of experts in a rich academic environment and in partnership with a community organization, will support the PI’s transition to independence as a clinical scientist who possesses unique skills and expertise to adapt evidence-based interventions within the context of pediatric cancer to meet the needs of children with cancer and their families.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract T cells play a central role in immunity and are of great therapeutic importance. Studies on T cell development provide direct insights into the pathophysiology of a wide variety of diseases and can facilitate adoptive T cell immunotherapies. Though a large effort has been made to identify pathways that regulate T cell differentiation, these studies have mostly focused on secreted proteins, cell surface receptors, and transcription factors known to directly drive developmental processes. Recently, we discovered that manipulation of an epigenetic regulator of lymphoid potential promotes the generation of mature, functional T cells from human induced pluripotent stem cells (iPSCs), demonstrating an important role for epigenetic modulators in T cell development and function. To date, the epigenetic regulation of T cell development remains elusive. In this application, we propose to test the hypothesis that iPSCs provide a platform to identify novel mechanisms regulating T cell development, and new insights into epigenetic regulations of T cell differentiation can be used to facilitate the generation of more robust iPSC-T cells for adoptive T cell immunotherapy. Previously, we have established a stroma-free differentiation system that faithfully recapitulates T cell development in culture. In Aim 1, we will perform small molecule screens at different stages of iPSC- T cell differentiation to discover new epigenetic regulators that can affect lymphoid specification and T cell maturation. We have provided a proof-of-concept by conducting a pilot screen using a library of small molecules that modulate the activity of epigenetic modification enzymes. The screen identified several epigenetic modulators, such as histone methyltransferase G9a, that promote T cell fate commitment. In Aim 2, we will transition our study to mechanistically examine how epigenetic regulations govern lymphocyte fate decisions via modulating chromatin structure. The impact of epigenetic modulators, including G9a, on lymphopoiesis will be probed in both iPSC-derived blood cells and zebrafish models. Lastly, Aim 3 will focus on using small molecule-mediate epigenetic modulation to facilitate the production of mature iPSC-T cells with enhanced function. Success in the proposed study will not only improve our understanding of the molecular mechanisms underlying the formation of immune cells but also open new avenues for stem cell-based immunotherapies. The career development award will allow me to develop new skills necessary to fulfill the proposed goals and foster my development into an independent investigator.

NIH Research Projects · FY 2025 · 2025-07

SUMMARY We request funding to purchase of an IVIS™ Spectrum 2 to be housed at the Rutgers New Jersey Medical School (NJMS) Advanced Preclinical Imaging Laboratory (APIL) which is part of the Rutgers University Molecular Imaging Core (RUMIC). The Spectrum 2 is a state-of-the-art optical imaging platform for in vivo imaging of live specimens ranging from cells to rodents. It will support 20 currently-funded projects of 8 major users and additional minor users. It advances research programs focused on parasitic and viral infections, pancreatic cancer, Alzheimer’s Disease, intestinal diseases such as colitis and improved methods for drug delivery. The investigators listed on this shared instrumentation proposal utilize optical imaging to study a wide variety of biological processes. One group of investigators uses it to detect and quantify drug distribution. A second group of investigators uses it to track disease progression in different tumor models. A third group of investigators uses it to detect dissemination of pathogens and quantify pathogen replication. The thermoelectrically cooled CCD camera in the Spectrum 2 enables use of a broad range of bioluminescent, fluorescent and chemiluminescent reporters, as well as Cerenkov luminescence. It attains improved quantum efficiency from a patented coating that captures more signal across the visible and near- infrared wavelengths. This enables highly sensitive, non-invasive 2D bioluminescence and fluorescence imaging which can be algorithmically reconstructed into 3D images. The improved signal-to-noise ratio provides high- quality images with excellent resolution. The Spectrum 2 has a standard high-throughput 22.5 cm field of view for simultaneous imaging of 5 mice, extendable to 10 mice with the optional 10-mouse manifold. Trans- illumination from underneath the samples enables detection of fluorescence even from deep tissue and epi- illumination aids imaging that requires higher throughput. The image analysis software is widely adopted, intuitive and user friendly. It removes autofluorescence and separately quantifies multiplexed fluorescent signals. Fluorescence and bioluminescence signals are quantified in 2D or reconstructed 3D space and viewed in the context of idealized anatomical structures. Accessories are designed to streamline imaging workflow and boost throughput for expedited data acquisition. In summary, the IVIS Spectrum 2 represents cutting-edge in vivo optical imaging that will propel research forward and lead to therapeutic advances to improve human health.

NIH Research Projects · FY 2025 · 2025-07

SUMMARY Staphylococcus aureus is an important Gram positive pathogen of the airway. It is of great concern due to its significant morbidity, mortality and antibiotic resistance, and has been acknowledged as a high priority target for new research from major international research and government bodies. The long term goal of our research is to understand the host-pathogen interactions between S. aureus and the host in the context of the airway infection. The objective of this proposal is to determine the signaling mechanisms and specific cell types behind the ability of S. aureus to evoke trained immunity in the airway epithelium. Our rationale underlying this proposal builds upon our significant in vitro and in vivo preliminary data that demonstrates only viable S. aureus is able to evoke trained immunity in the airway and this is driven by airway epithelial cells that demonstrate alterations in DNA modification, glycolysis and metabolism. Understanding these concepts in the context of infection will provide a greater understanding of host resistance mechanisms that may be adapted to new therapeutic concepts for at-risk patient populations for prophylactic use. Our specific aims investigate training of the innate immune system to S. aureus, with a focus on airway epithelial cells. In Aim 1 we will determine the factor of S. aureus that drives trained immunity and that sensing results in epigenetic changes to the chromatin architecture. Our goal for Aim 2 is to identify, using single cell sequencing, the specific epithelial subset that drives trained immunity and demonstrate its role through cell-specific knockout mice. This research is innovative as it combines next-generation sequencing approaches scRNA-seq and ATAC-seq with powerful mouse models to define the sensing and roles of airway epithelial subsets in trained immunity in the airway. The proposed research is significant, as it will further investigate our novel observation that airway epithelial cells are involved in trained immunity and will be the first study to attribute specific bacterial factors trained immunity. We will be able to further dissect this response to specific cell subsets. The results will have an important positive impact immediately because they will establish a better understanding of how the host innate immune system can be altered to better defend against this important pathogen, and long-term, because it lays the groundwork to potentially target specific aspects of the innate immune system to prevent infections in at-risk patient populations.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Acute kidney injury (AKI), characterized by a rapid decline in kidney function, is a growing health concern as it is significantly associated with increased morbidity and mortality, as well as reduced quality of life. In particular, vancomycin, an antibiotic commonly used empirically in hospitalized patients, is associated with a 2.45-fold increase in AKI risk. Despite its pronounced nephrotoxicity, the need to cover a broad spectrum of bacteria including potentially resistant strains often limits treatment choices to vancomycin. In a recent retrospective analysis by our research team, patients receiving the sleep medication, melatonin, along with vancomycin had a 63% lower incidence of AKI compared to those receiving vancomycin alone. Melatonin, a hormone that regulates circadian rhythms, is well established for its antioxidant activity. This indoleamine is preferentially taken up by the mitochondria, thereby eliminating free radicals and preserving mitochondrial functions. Melatonin likely also acts as an antioxidant by up-regulating nuclear receptor erythroid 2-related factor 2 (NRF2) target genes involved in the cellular defense system against oxidative stress. Notably, its antioxidant function is speculated to be also mediated through the activation of G-protein coupled melatonin receptors, MT1 (MTNR1A) and MT2 (MTNR1B). Therefore, to address an urgent need to identify novel interventions for management of vancomycin- induced nephrotoxicity, the proposed fellowship will investigate the mechanisms by which melatonin and related pharmacological agents protect against vancomycin toxicity in human kidney cells in vitro. The central hypothesis is that melatonin ameliorates antibiotic-associated kidney injury through coordinated activation of MT receptors and the NRF2-ARE signaling pathway. Aim 1 will investigate the role of melatonin receptor isoforms in mediating the renoprotective effect of melatonin and FDA-approved melatonin receptor agonists against vancomycin kidney toxicity. Immortalized and primary human proximal tubule epithelial cells (HK-2 cells and hPTECs) will be used to evaluate the changes in kidney injury (real-time apoptosis, mitochondrial health, and up-regulation of a proximal tubule injury biomarker), as well as other parameters, including receptor internalization and intracellular drug concentrations. Aim 2 will define the MT-NRF2 signaling gene network responsible for melatonin renoprotection by leveraging a stably transfected HK-2 cell line with disrupted NRF2 activity. RNA sequencing and qPCR will be used to dissect the transcriptomic profiles of genes modulated by the MT-NRF2 network. Consequently, elucidation of the renoprotective mechanisms will provide convincing evidence to re-purpose melatonin for a new clinical indication to prevent vancomycin nephrotoxicity in a cost- effective manner. Upon completion of the proposed studies and training, the Principal Investigator will be well- equipped to ultimately become an independent scientist and faculty member studying drug-induced renal injury at an R1 university.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT - OVERALL Until recently, Tuberculosis (TB) has been viewed as a disease that progresses over several discrete stages, principally consisting of a period of infection followed by either active TB disease or a latent state with the potential for reactivation. Similarly, Mycobacterium tuberculosis (Mtb), the causative agent of TB, has been viewed as a relatively stable bacterium with little genomic diversity, predictable causes of antibiotic resistance, and phenotypic uniformity both during culture and within its infected host. However, recent findings, many spearheaded by the members of this application, have begun to discover unexpected heterogeneity in TB disease states, host responses, the genotypes and phenotypes of the bacteria, and among the apparently clonal infecting population of Mtb. The premise for this program is that the heterogenous outcomes of TB infections and treatments are determined by the interplay between heterogeneous host-bacteria transcriptional and metabolic programs. Host and bacteria may be pre-programmed phenotypically or genetically to progress from TB infection to TB disease; and to do so rapidly or slowly; and, with or without extensive inflammation and lung damage. Immune tolerance, evasion or subversion may be another result of these interactions, which could lead to worsening disease and adverse treatment outcomes including relapse. Drug tolerance or resistance is another result of these interactions that may have widespread effects on treatment responses. Although Mtb-host and Mtb-drug interactions would seem to be unrelated, we will also study the possibility that immune and drug tolerant Mtb share a number of transcriptional and metabolic programs; and thus, also share some of the same vulnerabilities that could provide therapeutic targets. Consisting of 4 Projects and 3 Cores, this program will be accomplished in the following Specific Aims: 1) To determine the effects of bacterial and host heterogeneity on the manifestations, progression and consequences of close exposure to TB in the household, and of active TB. Addressed in Project 1: Bacterial and Host Determinants of Progression, Manifestations and Consequences of TB. 2) To uncover the immunological mechanisms underlying the diverse clinical outcomes in hosts infected with high and low transmission strains of Mtb. Addressed in Project 2: Immune Determinants of the Course of Mtb infection and Disease. 3) To define the host immune pathways that induce drug tolerance and identify potential routes to therapeutic intervention. Addressed in Project 3: Minimizing in vivo Drug Tolerance Induction in TB. 4) To define bacterial factors that contribute to the heterogeneous expression of drug tolerance and characterize links with adverse treatment outcomes. Addressed in Project 4. Drug Tolerance, Bacterial Heterogeneity and Adverse TB Treatment Outcomes.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Premium cigars, which were responsible for the cigar boom in the late 1990s, are now experiencing a resurgence. Through litigation, political forces have succeeded in protecting premium cigars from regulatory oversight via a series of court decisions. Ultimately, a court ruled to vacate FDA of its authority to regulate premium cigars in August 2023. The lack of evidence on the public health impact of premium cigars (e.g., effects on initiation and use among young people) played a major role in these court decisions. Given the paucity of evidence, the FDA commissioned the National Academies of Sciences, Engineering, and Medicine (NASEM) to evaluate the available evidence on premium cigars and to identify future research needs. As noted in the NASEM report, most data on cigar use has either been non-specific (i.e., does not distinguish between cigar types) or has focused on inexpensive, mass-produced cigars. The limited data on users of premium cigars suggest that they are distinct from users of other cigars and that their patterns of use also differ. The negative health effects and extent of addiction of premium cigars vary based on use patterns, therefore examining how premium cigars are used is essential for understanding their impact on public health. Research is also needed on factors that impact patterns of use, such as exposure to cigar marketing. Tobacco marketing is a risk factor for tobacco use in the general population—young people in particular—and marketing research is well-established for cigarettes and other cigars. Yet, research is limited for premium cigars. Premium cigars are not heavily promoted via traditional channels like other tobacco products (e.g., mainstream magazines), but are the focus of lifestyle magazines (e.g., Cigar Aficionado) and online marketing (i.e., websites and social media). Yet the content and impact of premium cigar messaging on perceptions and behavioral intentions in both users and vulnerable non-users (e.g., young adults), remains largely unexamined. The central objective of this multi-method study is to begin to address priority research gaps identified in the premium cigar NASEM report, and to serve as important precursor to a R01 proposal that comprehensively examines perceptions, patterns, severity of dependence, and influences (e.g., marketing) of premium cigar use over time. However, the findings from this R21 can also be used immediately to inform federal, state, and local decision making regarding premium cigars. Specifically, we aim to: 1) Understand consumer experiences and perceptions of premium cigars via focus groups; 2) document and characterize premium cigar marketing via a longitudinal content analysis; and 3) assess the effect of features from premium cigar advertisements on product perceptions and intentions via an online survey experiment.

NIH Research Projects · FY 2025 · 2025-06

Summary/Abstract Hepatitis C virus (HCV) causes a global epidemic with 71 million people infected. HCV is a blood-borne flavivirus that can cause cirrhosis and liver cancer and is responsible for ~400,000 deaths each year. Direct- acting antivirals (DAAs) can effectively cure HCV infection; however, most infected individuals are unaware of their infection status or only receive treatment after severe liver damage. Moreover, DAAs are cost-prohibitive and do not reach at-risk populations including people living in low- and middle-income countries or injecting drug users. Consequently, the number of yearly new infections (~1.4 million) exceeds the number of people receiving treatment (~650,000), thereby exacerbating the epidemic. Elimination of HCV by 2030 – a goal set by the World Health Organization (WHO) – will require multifaceted, decentralized public health prevention strategies. One such strategy calls for the development of point-of-care assays for active HCV infection, which is expected to accelerate the diagnoses of individuals who are most likely to transmit the virus and link those infected to care. As such, this proposal aims to develop a low-cost, instrument-free rapid antigen test for active HCV infection. We recently immunized mice with the HCV core antigen (HCVcAg), a protein secreted into the blood that can be detected almost simultaneously with HCV RNA during an active infection. Using flow cytometry, we sorted HCVcAg-specific B cells from immunized mice, then sequenced the antibody variable segments, and cloned and recombinantly expressed monoclonal antibodies (mAbs) that target the HCVcAg. In Aim 1, we will characterize the binding kinetics of the HCVcAg-specific mAbs. The mAbs with the most robust binding kinetics will then be used to generate a rapid antigen test. In Aim 2, we will determine the analytical and clinical sensitivities and specificities of the rapid antigen test using recombinant proteins, native viruses, and cryopreserved, de-identified serum/plasma previously collected from HCV viral load positive and negative patients and healthy controls. Our rapid antigen test is expected to have a sample-to-result processing time of <20 minutes and a maximum manufacturing price per test of <$4 USD. The successful completion of this project will lead to one of, if not the first, cost-effective, instrument-free rapid antigen test for the point-of-care diagnosis of active HCV infection.

NIH Research Projects · FY 2025 · 2025-06

Abstract B cells differentiate to produce antibodies that neutralize and inhibit viral entry aiding in host protection from infections. Following infection, B cells undergo CD4 T cell-dependent maturation and differentiation either in germinal center (GCs) or extrafollicular (EF) pathways within secondary lymphoid organs. B cells that have the potential to differentiate into antibody-secreting cells (ASCs) following secondary infections such as conventional memory B cells and CD11c+Tbet+ B cells are generated via both EF and GC reactions. Yet, the specific contribution of each of these B cell subsets to the generation and activity of anti-viral antibodies upon re-infection remains unclear. CD11c+Tbet+ B cells (herein referred to as Tbet+ B cells) are a distinct B cell subset that differentiate following viral infections as well as autoimmunity in mice and humans. Emerging studies have shown that in murine and human infections, Tbet+ B cells are critical drivers of protective antibodies. Tbet+ B cells are capable of differentiating into class-switched autoantibody-secreting cells upon stimulation by cytokines and TLR agonists. Additionally, these B cells have elevated expression of MHCII and secrete inflammatory cytokines such as IFNg and IL-12 following stimulation, suggesting a role in effector T cell activation. The development of Tbet+ B cells in a primary infection has been well elucidated, but their differentiation and contribution to host protection following secondary viral infection is poorly understood. Our new data and published studies demonstrate that Tbet+ B cells develop from the GC response, but mainly arise from the EF pathway following a primary viral infection. However, whether Tbet+ B cells that arise from GC or EF origin have different functions upon re-infection is unknown. Currently, we found that following the resolution of a viral infection both the GC and EF-derived Tbet+ B cells persist in the spleen. However, upon re- infection, the GC population appears to form secondary GCs or differentiate into ASCs, while the EF maintains a Tbet+ B cell phenotype indicating a role for T cell-mediated immunity. We hypothesize that the developmental origin of Tbet+ B cells influence their differentiation and function following secondary viral infection. Accordingly, our study will dissect how GC and EF origin impact transcriptional regulation and localization of Tbet+ B cells. We will utilize an allotypic transfer system to discern the protective contributions of Tbet+ B cells versus memory B cells following re-infection. We expect this work to reveal a novel mechanism of Tbet+ B cell regulation that critically impacts the host response following infection. Ultimately, this work should provide insight into new therapeutic targets to boost antibody responses and/or effector CD4 T cell responses in immunization strategies or during secondary infection or dampen harmful humoral responses during autoimmunity.

NIH Research Projects · FY 2026 · 2025-06

ABSTRACT Chlorine (Cl2) is a reactive gas that can cause severe lung damage. It has been used as a chemical warfare agent since World War I and recently has been used on a large scale in the Syrian civil war. It matches the criteria of a chemical of concern as defined by the countermeasures research program. In addition to its importance as a chemical warfare agent Cl2 injury is often seen in accidental exposures. The major route of Cl2 exposure is through inhalation and thus, most of the morbidity and mortality that results is due to respiratory complications. Signs and symptoms of Cl2 exposure include cough, choking, irritation and burning in the throat and upper airway, and general difficulty breathing. These symptoms typically display within the first 24 hr following exposure. In the long term, Cl2-mediated lung injury can result in a range of pathologies. The early initiation of injury and the varied long-term sequelae indicate that it is important to intervene within the first 24 hr. Therefore, we have developed a mouse model of inhaled Cl2 exposure that focuses on the first 24 hr for the purposes of testing medical countermeasures. Upon inhalation, Cl2 partitions into the lung lining fluid where it forms hydrochloric and hypochlorous acids and can react with the respiratory mucosa leading to lipid oxidation, DNA damage and cytotoxicity within both pulmonary epithelial and endothelial cells. These reactions result in surfactant dysfunction, loss of epithelial barrier integrity and consequent inflammatory activation. A key step in the inflammatory process is the recruitment of pro-inflammatory activated macrophages. We propose that inhibiting the recruitment and activation of pro-inflammatory macrophages will mitigate Cl2 toxicity in the lung. Electrophilic fatty acids (EFAs) are a class of compounds that have been found to inhibit inflammation in a variety of circumstances. Previously, we have found that the naturally occurring EFA nitro-oleic acid (OA-NO2) inhibits macrophage activation and recruitment in bleomycin-mediated lung injury. In preliminary studies, we have seen this effect in Cl2 injury. It is our contention that by limiting macrophage activation, one can reduce the consequent inflammatory activation and its associated injury. It is our hypothesis that systemic delivery of EFAs post Cl2 inhalation will reduce macrophage recruitment and activation leading to mitigation of lung injury. There are a wide variety of substitutions that can be made upon EFAs that will alter their target specificity and pharmacokinetics. Within the first aim of this study, we will pre-screen up to 55 EFAs for their ability to inhibit macrophage activation in vitro with a view to identifying the most effective potential treatments of Cl2 toxicity in the lung. In our second aim we will examine how i.p. administration of five lead compound EFAs each with a different structural alteration can lead to pro-inflammatory macrophage inhibition and alleviation of injury. These studies will yield mechanistic information as to the role of macrophage recruitment in Cl2-induced lung injury and how EFAs can alter pro-inflammatory activation. As well as assessing the potential of EFAs as medical countermeasures for Cl2 induced lung injury.

NIH Research Projects · FY 2026 · 2025-06

ABSTRACT Mycobacterium tuberculosis (Mtb) infection is a global health pandemic claiming 1.5 million lives each year. Current tuberculosis (TB) therapies are challenged by drug resistance, characterized by mutations that render approved drugs increasingly less effective. PZA is a critical component of first-line TB therapy. It allows for shorter treatment regimens and is unrivaled in its efficacy against non-replicating, persister Mtb populations, which decrease the chance of resistance and relapse respectively. If the past predicts the future, the success of many short course and ultra-short course TB therapies may depend on the presence of PZA or other POA releasing prodrugs as part of the treatment regime; thus, the development of a replacement for PZA in the setting of PZA resistance should be among the highest priorities of modern TB drug development. POA itself is relatively polar and has poor penetration into the bacterium. However, PZA crosses the Mtb cell envelope via passive diffusion and is then activated by PncA, an amidase present in the cytosol, to release POA. The amide structure of PZA thus allows the transport and delivery of POA into the cytosol. Unfortunately, PZA resistance is increasing, with approximately half of all isoniazid and rifampin resistant Mtb also showing PZA resistance, usually due to inactivating mutations in the pncA gene. Fortunately, Mtb contains a wide variety of amidases other than pncA that can activate antitubercular compounds which contain a central amide bond. This proposal focuses on the potential for using Mtb amidases as a new class of antitubercular prodrug activators that can circumvent existing forms of resistance. We have identified over 150 Mtb genes that are likely to encode amidases. We have also shown that, like PncA, the Mtb amidase AmiC is involved in the activation of several experimental antitubercular compounds, although AmiC and PncA have different substrate specificities. Conversely, we have shown that approximately 20% of amide containing compounds present in libraries with validated antitubercular activity undergo rapid intrabacterial amide hydrolysis. Thus, different Mtb amidases can specifically activate prodrugs, and diverse antitubercular compounds can be activated by intrabacterial amide hydrolysis. We propose to characterize the diverse set of Mtb amidases and the compound structures that they hydrolyze into paired acids and amines. We will then use this knowledge to rationally design new POA-amide prodrugs that readily diffuse into Mtb where they can then become activated by intrabacterial amidases, releasing POA. We will then confirm that these compounds have activities similar to PZA but are effective against both PZA susceptible and resistant Mtb. Finally, we will perform hit-to-lead optimization and confirmatory pharmaco-kinetic studies in a mouse model.

NIH Research Projects · FY 2025 · 2025-06

Abstract From Original Submission Epilepsy is a severe neurological disease affecting more than 65 million people worldwide and is characterized by unpredictable abnormal electrical discharges resulting in recurrent seizures. About one third of patients with epilepsy suffer from intractable seizures that do not respond to anti-seizure medications (ASMs). Neurosurgical interventions and neurostimulator devices are useful options for only a fraction of patients with drug-refractory seizures, underscoring the urgent need to develop new therapies. One strategy with considerable promise is to engraft new neurons to provide enhanced GABAergic inhibition in an activity-dependent manner. However, use of fetal neurons for cell therapy is associated with practical and ethical issues. Therefore, to overcome such hurdles, in our previous studies, we pioneered the transplantation of human pluripotent stem cell (hPSC)- derived medial ganglionic eminence (MGE)-type human cortical interneurons (cINs) into epileptic mouse brains and demonstrated their integration into dysfunctional circuitry, accompanied by the suppression of seizures and comorbid behavioral abnormalities. Furthermore, more recently, we have determined the optimal stage of human cIN differentiation to ensure maximal integration into host circuitry as well as safety without risk of tumor formation, and developed a method to efficiently generate these safe and highly migratory populations of synchronized early postmitotic cINs from hPSCs in large quantities, bringing cell therapy for epilepsy one step closer to reality. Furthermore, we have successfully tested the efficacy of human early postmitotic cINs in 2 different models of temporal lobe epilepsy (TLE), observing >80% of seizure reduction. With these strong previous studies, now we are ready to embark clinical translation of this novel and restorative therapy for epilepsy patients with limited options. Thus, in this proposed study, we will scale up production of synchronized early postmitotic cINs that are optimal for grafting under cGMP condition. For added safety, we will utilize well- characterized HLA-edited hypoimmunogenic iPSCs to minimize the need for immunosuppression for off-the- shelf use of human cINs. We will also extensively analyze the produced early postmitotic cINs’ phenotype, efficacy, safety, tumorigenesis and biodistribution to seek IND approval. Once we obtain IND approval, we will do a first-in-human clinical trial of early postmitotic cIN grafting with a primary goal of safety analysis, while also checking efficacy as a secondary measure. This will be done in patients with intractable TLE who are candidates for resection while they undergo intracranial EEG to identify the seizure focus without additional invasive steps. Completion of these studies is pivotal for translating this experimental therapy into a viable therapeutic strategy for intractable epilepsy.

NIH Research Projects · FY 2025 · 2025-05

Abstract/Project Summary We are seeking funding to purchase a Refeyn Two-MP Mass Photometer, a cutting-edge instrument that will revolutionize our research capabilities. This instrument enables quantitative label-free single-particle mass measurement of biomolecules in solution, resolving a broad range of mass species (30 kDa to 5 MDa) across various oligomeric states, quantifying protein molecular interactions, determining stoichiometry, and optimizing conditions for functional assays and macromolecular assembly. Its integration will significantly enhance structural, biochemical, and biological research by NIH-funded researchers at Rutgers University and Rutgers Health. The instrument will be housed at the Mass Spectrometry core facility within the Proteomic building at Rutgers University and Robert Wood Johnson Medical School (RWJMS). It will be the first of its kind across Rutgers University and Rutgers Health campuses, filling a critical void in available technologies and benefiting numerous investigators. Core users will include NIH-funded and junior investigators’ research laboratories, with accessibility extended to other researchers. The instrument's unique ability to quantify mass with high resolution and accuracy under native conditions with minimal sample volume and concentrations, as well as its capacity to measure interactions in the nanomolar concentrations, make it an invaluable tool for screening samples for structural and functional studies. Recent advancements in the microfluidics attachment to the Two-MP enable the quantitation of weak molecular interactions at micromolar concentrations, further enhancing its utility. The instrument’s capabilities extend beyond conventional methods, potentially replacing or complementing techniques such as size exclusion chromatography, multiangle light scattering, electrophoretic mobility shift assays, and protein interaction assays. We have strong institutional support from the Rutgers Health Chancellor, RWJMS Dean, and Rutgers University to ensure support staff and space in the core facility, and service contracts. The operation of the instrument is user-friendly, akin to the nanodrop instrument, requiring minimal training. A support staff at the core facility will oversee routine cleaning, maintenance, and coordination of usage and fees. A technical expert committee composed of local members with strong expertise in single-molecule optical methods will provide consultation on experimental design and advice on data interpretation. An advisory committee has been formed to ensure proper and efficient use of the instrument. The anticipated outcomes of this acquisition are significant, including bolstering current NIH-funded projects and empowering junior faculty to explore previously inaccessible research avenues. The instrument is expected to catalyze new collaborations and discoveries across the basic, clinical, and applied research missions of Rutgers and to accelerate biomedical discoveries widely.

NIH Research Projects · FY 2025 · 2025-05

ABSTRACT Autism Spectrum Disorder (ASD) and other neurodevelopmental disorders (NDDs) are more prevalent in males, but the molecular mechanisms behind this sex bias are unclear. This proposal investigates how hormones and sex-specific signaling interact in developing behavioral deficits, using mouse models of Coiled-coil and C2 domain containing 1a (Cc2d1a) loss-of-function. Human biallelic null mutations in CC2D1A cause intellectual disability (ID), ASD, and seizures. Male Cc2d1a-deficient mice display behavioral deficits that recapitulate features of ASD (reduced cognition and sociability), which are absent in females. CC2D1A is a signaling scaffold that modulates cyclic AMP (cAMP) signaling, crucial for synaptic plasticity and learning and memory. Our previous research showed sex-specific disruptions in cAMP signaling in the hippocampus of Cc2d1a-deficient males, that could be rescued only in males by modulating cAMP levels. This indicates that CC2D1A loss leads to sex-specific brain signaling deficits affecting cognition and sociability in males only. We hypothesize that gonadal hormones establish these sex-specific signaling deficits upstream of CC2D1A function leading to distinct male and female behavioral outcomes. However, gonadal hormones can act at different developmental stages to establish sex-specific mechanisms with organizational role early after birth and activational (acute) roles after puberty. Early postnatal lethality in global Cc2d1a knockout (KO) mice previously hindered these studies as the two timepoints could not be studied in the same animal. We recently generated a new hypomorphic mouse strain with a 90% reduction in CC2D1A levels that allows hormone modulation at various times during development. Our expertise in CC2D1A function complemental with advice from an expert neuroendocrinologist perfectly position us to conduct the proposed research. In Specific Aim 1, we will determine whether gonadal hormones cause male-specific behavioral deficits in Cc2d1a-deficient mice through organizational or activational action. We will treat female pups with estrogen postnatally to mimic the male testosterone surge and reduce estrogen/testosterone levels in adults via ovariectomy or orchidectomy. In Specific Aim 2, we will probe the molecular mechanism underlying how sex hormones establish sex-specific intracellular signaling in neuronal cultures from Cc2d1a KO and hypomorph mice. This combined approach will These studies will explore a new molecular mechanism for sex-specific cognitive deficits, also informing whether therapeutic approaches should be sex-specific.

NIH Research Projects · FY 2025 · 2025-05

PROJECT SUMMARY/ABSTRACT This integrated research and training plan will prepare Dr. Subramanian, an attending in a dental school- and hospital-based residency program, to become an independent NIH-funded dentist-scientist with expertise in non-opioid pain-mitigative strategies during post-surgical recovery, following prevalent surgeries such as impacted third molar removal. Annually, over 3.5 million predominantly young and healthy individuals undergo out-patient wisdom tooth extractions. A vast majority of these individuals are prescribed postoperative opioid pills, a practice that often introduces opioids to opioid-naïve patients and poses a risk of future repeat use. Pain and limitation to mouth opening remain the two most serious consequences during the week of post-surgical recovery. This innovative proposal explores the ability of a novel local anesthetic injection (the temporo-masseteric nerve block or ‘TMNB’) developed by Dr. Subramanian and team, that ‘numbs’ the jaw closer muscles to reduce post-surgical pain and ease mouth opening during the post-surgical week. The TMNB has shown early promise in providing sustained pain and jaw muscle spasm relief post-dental surgical procedures and in alleviating chronic jaw muscle pain. Using the TMNB is expected to complement pain relief from non-opioid analgesics, and, secondarily, reduce resorting to opioids, mitigating the prescription opioid crisis. Under the guidance of a strong team of NIH-funded clinical trialists, Dr. Subramanian will accomplish two key training goals that integrate with the specific aims of her research plan 1) Gain experience in designing, executing, and analyzing clinical trials, and 2) Learn qualitative research methods and analysis. Dr. Subramanian’s training activities will be complemented by an innovative research plan that will provide experiential and practical exposure. She will test the feasibility and potential efficacy of the TMNB in reducing post-surgical pain and jaw functional limitation compared to conventional treatment in a triple blind, placebo- controlled pilot phase II trial. Per this study design, 80 subjects undergoing impacted lower third molar extractions under sedation will be randomized to receive the TMNB injection/ saline if they develop significant jaw muscle pain after surgery. In addition to chairside outcome assessment, patient-reported outcomes will be collected remotely on a daily basis for a week. All participants will receive identical guidance for pain medications usage. At the end of study, Dr. Subramanian will conduct qualitative interviews with subjects who were previously randomized to TMNB/placebo to understand potential patient-centered strategies to describe the TMNB experience, understand post-operative recovery and guide oral analgesic use during the postoperative period. This research will be critical for designing rigorous future clinical trials to validate the TMNB intervention. This innovative K23 Mentored Patient Oriented Career Development Award responds to NIH high priority HEAL topics including the exploration of non-opioid strategies to improve pain control and ultimately develop guidelines to improve pain management and reduce reliance on opioids.

NIH Research Projects · FY 2026 · 2025-05

Summary Aging is a critical risk factor for heart disease. Cellular senescence, characterized by cell cycle arrest, resistance to apoptosis and the senescence-associated secretory phenotype (SASP), occurs in many cell types, including cardiomyocytes (CMs). Senescence precipitates the aging process in surrounding cells and the organ as a whole through paracrine mechanisms. Generalized autophagy is decreased during aging in many organs, including the heart. This decrease causes deterioration of cellular quality control mechanisms, facilitates aging and negatively affects lifespan in animals, including mice. Mitophagy, a mitochondrion- selective form of autophagy, plays an essential role in mediating mitochondrial quality control mechanisms. We have recently shown that alternative mitophagy, an unconventional form of mitophagy that does not rely on the conventional mechanism of autophagy, plays a major role in mitochondrial quality control during myocardial ischemia when conventional autophagy/mitophagy is inactivated. Although suppression of generalized autophagy could promote senescence, senescent cells are metabolically active and rely on mitochondria; thus, alternative mitophagy may be stimulated in order to maintain mitochondrial function. Therefore, the major goal of this project is to elucidate the role of autophagy and mitochondrial quality control mechanisms, including mitophagy, in senescence of the heart and the CMs therein. Our overall hypothesis is that aging- and stress- induced suppression of autophagy promotes senescence in CMs. Suppression of general autophagy leads to the accumulation of YAP, the terminal effector of the Hippo pathway, and consequent stimulation of alternative mitophagy, which are required for the survival and the SASP of senescent CMs, thereby promoting cardiomyopathy. To prove our hypotheses, we will: Aim 1: Test whether downregulation of autophagy directly induces senescence in CMs through YAP-dependent mechanisms. Aim 2: Test whether alternative mitophagy promotes survival and the SASP in senescent CMs. Aim 3: Test whether YAP and alternative mitophagy mediate CM senescence and the cardiac aging phenotype in mouse models of aging. Using mouse models of cardiac senescence in which general autophagy is downregulated, the critical involvement of YAP activation and alternative mitophagy in senescence-mediated cardiac dysfunction will be demonstrated by conducting CM- and senescent cell-specific genetic manipulation of YAP and alternative mitophagy together with fate mapping of p16High senescent cells. Expression of specific diphtheria toxin fragment A in p16High cells also allows selective killing of senescent cells. We will show that unique molecular mechanisms more strongly activated in senescent CMs than in non- senescent cells, namely YAP and alternative mitophagy, promote the survival and the SASP of senescent CMs, thereby mediating the development of cardiomyopathy. Our study will provide important clues for the development of novel senolytics to alleviate cardiac dysfunction induced by CM senescence and aging.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY/ABSTRACT Blunts – cigars with the tobacco filling replaced or mixed with cannabis – present substantial health risks to their users given exposure to harmful chemicals from combustion and the high addictive potential from nicotine in the cigar wrap and tobacco. Blunts are predominantly used by young adults (YAs) and groups who go on to suffer disproportionately from tobacco-attributable death and disease, chiefly people who identify as Black/African American (B/AA). Therefore, B/AA YAs are at greatest risk for the health effects of blunt use. In the United States, tobacco policies have become increasingly restrictive while cannabis policies have become increasingly permissive. In this rapidly changing context, it is important to understand how blunt use is impacted by both tobacco and cannabis policies. However, no studies have evaluated these impacts. There is a critical need to understand the impact of evolving cannabis and cigar policies on people who use blunts, particularly B/AA YAs, to promote public health. Aligned with National Institute on Drug Abuse research priorities (NOSI NOT-DA-22-003), the overall objective of the proposed study is to determine the impact of policies relevant to blunt use on health outcomes. I will build upon analyses of national data and focus group data (K99 phase) to construct a hypothetical choice experiment (R00 phase) to predict potential impacts of intersecting tobacco and cannabis policies on blunt use in B/AA YAs. The first aim is to detect the impact of state cannabis and tobacco policies relevant to blunt use on B/AA and non-B/AA YAs through secondary analyses of Nielsen convenience store data and Population Assessment of Tobacco and Health Study data. The second aim is to understand multi-level determinants of blunt use and policy impacts through focus groups with B/AA YAs. The third aim is to assess the impact of proposed cannabis and tobacco policies to reduce blunt use among B/AA YAs through a discrete choice experiment. This study will provide timely evidence on an understudied pattern of cannabis and tobacco use (blunt use) impacted by rapidly-evolving tobacco and cannabis laws. Results are expected to have substantial positive impact by 1) directly informing policy interventions to support public health, 2) identifying determinants of policy impacts on blunt use, and 3) introducing unique scientific approaches to a budding field of cannabis regulatory science. A comprehensive training plan is proposed to ensure the implementation of study aims, with objectives focused on: expanding data analytic skills for policy evaluation; obtaining experience with experimental methods to inform and evaluate policy attributes; extending knowledge in core substantive areas of cannabis use and policy; and developing leadership and professional skills relevant to tobacco and cannabis regulatory science.

NIH Research Projects · FY 2026 · 2025-05

Project Summary/Abstract An increasing percentage of youth in the US report inadequate sleep and sleep problems, which negatively impact health and daily living, leading to the development and worsening of psychiatric and non-psychiatric health conditions. Individuals with ADHD suffer from sleep problems at higher rates compared to the general population, with sleep disturbances one of the most frequent adverse effects of stimulants, the primary ADHD treatment. Despite inadequate evidence on safety, a variety of medications are prescribed to treat sleep problems in youth. This proposal focuses on five commonly prescribed medications for sleep problems: trazodone, benzodiazepines, nonbenzodiazepine sedative hypnotics, alpha-blockers, and hydroxyzine. None of these are FDA-approved for pediatric insomnia. The safety of pharmaceuticals prescribed for sleep problems in youth is vastly under-researched, hampering treatment recommendations. Medications commonly prescribed for sleep disorders have been linked to risks of suicide, unintentional overdose, cardiovascular events, and hallucinations. However, these adverse outcomes have not been rigorously examined in youth. Further, risks may be magnified in stimulant-treated youth with ADHD, given independent risks of stimulants and of having ADHD. The proposed project is motivated by high rates of sleep problems and sleep medication use in youth, especially among those with ADHD, combined with a paucity of drug safety information and the potential for magnified risks in youth with ADHD. In this proposal, we will provide the first estimates for the safety of prescription sleep medications in children, adolescents, and young adults with ADHD. The study team will use two large, national administrative claims data sets (2001-2023) covering over half million privately and publicly insured youth with ADHD starting one of the study medications. We will evaluate nationwide age-specific patterns of prescription sleep medication use and dose-escalation in youth with ADHD (Aim 1) and determine the extent to which prescription sleep medications increase the risk of a) suicide attempt, b) unintentional overdose, c) cardiovascular complications, and d) hallucinations and whether stimulant treatment or ADHD magnify the risk of these harms in youth (Aim 2). Lastly, we will use novel signal detection methods to discover and evaluate new, biologically plausible adverse outcomes of prescription sleep medications in youth with ADHD (Aim 3). This research will yield new mechanistic insights into drug safety and drug-stimulant and drug-ADHD interactions, thus producing novel, generalizable, and actionable risk estimates of serious harms from prescription sleep medications in youth with ADHD. Our findings will enable patients, caregivers, and clinicians to make individualized evidence-based decisions concerning pharmacological management of sleep problems in youth. The proposed methods offer a valid, timely, and effective approach to yield clinically relevant evidence and support our long-term goal of generating rigorous safety data on sleep medications in youth with comorbid mental illness.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY/ABSTRACT Multiple forms of psychopathology involve problems with resisting temptations and urges. Experience sampling (ES) methodology allows the assessment of multiple dissociable facets of temptations, including their intensity and whether they are successfully resisted. Despite their relevance to multiple forms of externalizing psychopathology and certain internalizing conditions, studies applying the ES approach to urges and their resistance have not to date been integrated into research on the structure of psychopathology. Rather, existing ES studies have focused narrowly on specific disorders in isolation (e.g., examining craving for one specific drug while ignoring the multitude of other temptations that the person experiences). Hence, it is still unknown whether individuals with relevant psychopathology display a broad pattern of differences in temptations and their resistance beyond the abused substance or behavior that is the focus of that disorder. Furthermore, it is unknow to what extent observed findings are specific to these individual disorders or reflect transdiagnostic features that contribute to the high comorbidity of conditions across broader dimensions (spectra and superspectra) of psychopathology. We propose to test the hypothesis that the broad ability to successfully resist temptations and urges across temptation classes is associated with the broad externalizing superspectrum, and preferentially its underlying disinhibition spectra as defined by the Hierarchical Taxonomy of Psychopathology (HiTOP). Additional associations are expected to arise between reduced temptation resistance success for specific temptation classes and narrower symptom domains associated with the externalizing and internalizing spectra (e.g., binge eating, nonsuicidal self-harm among internalizing symptoms). We further test genetic and environmental influences on temptation strength and resistance ability with a focus on polygenic scores, adverse childhood experiences and childhood neighborhood deprivation variables that have previously been related to externalizing. Finally, we aim to determine which resistance strategies are the most successful within and across different psychopathology domains (which has direct implications for interventions). To address these issues, we will study 1200 subjects (with enrichment for psychopathology) who will complete two weeks of experience sampling and broad measures of externalizing and internalizing. Because, our ES regimen collects data on the types of temptations and urges, their subjective strength, the goal of the urge (pleasure/relief from negative feelings), the level of conflict with other goals, whether the person attempted to resist the temptation, the resistance strategy, and resistance success, it becomes possible to test the extent to which differences in each of these facets are related to both narrow and broad dimensions of psychopathology. As the most comprehensive transdiagnostic assessment of temptations in psychopathology, the dataset will become a shared resource that allows testing of novel hypotheses about temptations and their resistance in mental health.

NIH Research Projects · FY 2026 · 2025-04

Project Summary: Systemic lupus erythematosus (SLE), the predominant form of the disease, represents a chronic autoimmune disorder characterized by the production of autoantibodies from hyperactive immune cells, along with pervasive inflammation. This immune dysregulation culminates in severe tissue damage to vital organs such as the kidneys, heart, joints, and skin. Despite significant advancements in SLE treatment over the past five decades, encompassing the use of immunosuppressive agents, numerous complications endure, including inflammation, osteoporosis, cardiovascular issues, acute renal failure, and lupus nephritis, perpetuating the burden of disease. To confront these enduring challenges, monoclonal antibodies (mAbs) targeting CD20 or CD19 antigens on B cells, such as rituximab, ocrelizumab, obexelimab, and obinutuzumab have been developed. These mAbs make a “low affinity” immunologic synapse with natural killer (NK) cells and a high affinity connection with B cells, instigating B cell depletion through antibody-dependent cell-mediated cytotoxicity (ADCC). However, the low affinity interaction of mAbs with NK cells poses a critical obstacle, leading to inefficient clearance of B cells, including memory B cells, in peripheral blood and tissues, contributing to rapid disease relapse. To bridge this gap in treatment efficacy, our proof-of-concept investigation endeavors to pioneer a new approach for SLE therapy. The objective of this study is to engineer a Trispecific Killer Cell Engager (TriKE), designed to engage and activate NK cells with high affinity, thereby facilitating efficient clearance of pathogenic immune cells in SLE. To our knowledge, the utilization of "high affinity" NK cell engagers for SLE therapy represents uncharted territory in medical research, signifying a technically innovative strategy. Leveraging our expertise, our laboratory has successfully isolated a unique single-domain antibody with “high affinity” and specificity for the CD16a receptor on NK cells. Capitalizing on this breakthrough, we intend to construct a TriKE that can bind to three target antigens, empowering NK cells to effectively identify and clear pathogenic immune cells. Through rigorous evaluation using peripheral blood from lupus patients, the efficacy of TriKE in eliminating pathogenic immune cells will be meticulously examined. Rituximab and obexelimab will serve as benchmark controls in our comparative analyses. Validation of NK cell activation and subsequent target immune cell death will be achieved through the quantification of NK cell degranulation, along with the release of granzyme B and perforin. In summary, our focus lies in the eradication of autoreactive immune cells. By eliminating these aberrant cells, we aim to offer not just transient symptomatic relief, but the potential for enduring alleviation for a majority of patients and even the possibility of a cure for select individuals. This transformative approach holds promise for revolutionizing the landscape of SLE treatment, offering hope for improved outcomes and enhanced quality of life for those affected by this debilitating condition.