Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2025 · 2026-02

PROJECT SUMMARY Cardiac development is a complex process that occurs early during embryonic development and requires precise regulation. Early cardiac development, especially of the endocardial and myocardial lineages, is implicated in the development of congenital heart diseases (CHDs). Unfortunately, despite CHDs being the most frequent birth defect affecting approximately one in 100 live births, very few mechanisms are understood about many different forms of CHDs. Proper cardiac development requires precise post-transcriptional regulation, such as alternative splicing, translation initiation, and mRNA modification. These functions are mediated by RNA-binding proteins (RBPs), and unsurprisingly, mutations to many RBPs are implicated in cardiac development and formation of CHDs. One such RBP is DDX3X, which is known from clinical evidence to have implications in CHDs: patients with mutations in DDX3X exhibit DDX3X syndrome, marked by neurodevelopmental disorders and increased risk of CHDs. However, DDX3X has not been well-studied to date in the context of heart development. My proposed research will study the dosage-sensitive effects and mechanisms of DDX3X in the endocardial lineage during cardiac development. In Aim 1 of this proposal, I will determine the effects of reduced DDX3X levels on endocardium formation and function. Using transgenic mice, I will conditionally delete DDX3X in male and female mouse endocardium. Expression level of DDX3X in the endocardium will be quantified using immunofluorescent imaging and flow cytometry. I will then characterize resulting phenotypes by examining phenotypic onset, structural and functional consequences, and cellular and molecular consequences in the heart using a combination of brightfield and immunofluorescent imaging, weight measurements, and echocardiograms. In Aim 2 of this proposal, I will identify the molecular mechanisms of DDX3X in the endocardium. Using immunofluorescent imaging, I will characterize the subcellular localization of DDX3X in the endocardium. I will then identify the direct regulatory network of DDX3X in the endocardial lineage using RNA-seq and Ribo-seq. Targets will be validated against pre-existing eCLIP data as well as in an in vitro system of human pluripotent stem cell-derived endocardial cells. The results of this research will contribute to our understanding about RNA biology, gene dosage, development and disease.

NIH Research Projects · FY 2026 · 2026-01

Project Summary Alzheimer’s disease (AD) and Alzheimer’s disease-related dementia (ADRD) and Parkinson’s disease (PD) are chronic neurodegenerative disorders characterized clinically by cognitive impairment and neuropathologically by abnormal protein accumulations (Aβ and α-Synuclein). Recent advances in understanding transcriptomic changes in AD and PD have focused on specific cell subpopulations and brain regions, such as microglia in the parahippocampal gyrus in AD and dopaminergic neurons in the substantia nigra in PD. Although significant, this narrow focus has diverted attention on other cell types and brain regions implicated in those diseases. Recognizing this shortfall, our project broadens the scope of studied cell subpopulations and brain regions with a special emphasis on deciphering impaired cellular cross-talk, detecting perturbed spatial interactions with morphological markers, and identifying gene regulatory programs driving AD and PD progression. In our previous work, we have established two resources through single-nuclei profiling. The first is a cross-disorder atlas that includes data from the prefrontal cortex of 1,494 donors with seven major brain-related disorders, including AD, PD, and other dementia. The second is a PD-specific atlas derived from 100 PD donors, encompassing five different brain regions. Our current proposal aims to leverage data from both AD/ADRD and PD datasets to systematically describe the differences in the transcriptional landscape of those diseases and augment the PD-centric atlas by integrating epigenomics and spatial transcriptomics. In Aim 1, we will characterize shared and distinct transcriptional changes between AD, PD and related dementia and identify cell types with significant proportional shifts associated with the disease progression. For the prefrontal cortex, included in both previously generated resources, we will conduct detailed comparative analyses between those diseases with a special focus on the interplay between AD and PD neuropathology measured by the Braak scoring system, i.e. accumulation of neurofibrillary tangles and Lewy bodies. Aim 2 focuses on developing an atlas of brain regulome changes in PD, which will pinpoint altered epigenetic regions leading to disruptions in chromatin structure. Utilizing both epigenomics and transcriptomics data will help us uncover enhancer-driven gene regulatory networks and model how these networks shape the transcriptome during disease progression by manipulating key epigenome regulators and related TFs. By integrating epigenome data with genome-wide association studies on PD, we will identify cell subtypes with increased PD heritability and refine the mapping between risk loci and the causal genes. Aim 3 will utilize integrative spatial genomic assays to detect differential cell-to-cell interaction networks, pointing to the gain or loss of ligand-receptor relationships and spatial relationships with morphological markers. Collectively, these studies will enable us, at unprecedented resolution, to explore brain region- and cell-type-specific transcriptomics and epigenomics responses in PD and distinguish them from those employed in AD and ADRD.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Alzheimer’s disease (AD) is a progressive neurodegenerative disease affecting more than 50 million people worldwide, but the mechanisms involved in disease development remain poorly understood. Genome-wide association studies have uncovered numerous loci associated with disease, but identifying causal variants and genes remains a challenge. Studies have revealed that AD risk variants are enriched in active enhancers of human monocytes, macrophages, and microglia, suggesting many of these variants act by disrupting gene expression specifically in myeloid cells. In addition, many genes implicated in AD risk are highly expressed in myeloid cells, strongly implicating these cell types in the etiology of AD. By integrating human genetics with epigenomic and transcriptomic datasets from human myeloid cells, we identified a myeloid cell enhancer containing an AD-associated functional variant on chromosome 11, and two putative target genes of this enhancer, embryonic ectoderm development (EED) and phosphatidylinositol binding clathrin assembly protein (PICALM). EED is an essential subunit of the polycomb repressive complex 2 (PRC2) known to function in regulation of gene expression and clearance behavior in mouse microglia, but it remains relatively unstudied in the context of AD in human cells. PICALM is an adaptor protein known to function in endolysosomal pathway and autophagy, but its role in microglia remains unknown. The overall goal of this proposal is to understand how our nominated AD risk enhancer influences the expression of its two target genes PICALM and EED, and to test the hypothesis that these two potential causal genes regulate microglia functions downstream of TREM2. In Aim 1, I will determine the role of the candidate AD risk enhancer in regulating the expression of PICALM and EED and human microglial cell function by combining CRISPR gene editing in human induced pluripotent stem cells (iPSCs), microglial differentiation protocols, and xenotransplantation methods involving direct injection of microglia precursor cells into the mouse brain. I will delete the candidate enhancer region and perform transcriptomic and epigenetic profile of edited microglia via RNAseq and ATACseq in addition to microglia functional assays. To assess the in vivo consequences of deleting the risk enhancer in microglia, I will transplant the edited microglia into the brains of wildtype and 5xFAD humanized mice and characterize transcriptomic profile of these microglia via snRNAseq analysis and perform immunohistochemistry. In Aim 2, I will investigate how PICALM and EED regulate human microglia function, namely TREM2 mediated efferocytosis, or phagocytic pathway. I will determine the subcellular localization of PICALM and EED using biochemical and imaging techniques and test the role of these two genes in microglial functions downstream of TREM2 signaling such as actin rearrangement, phagosome formation, and downstream kinase signaling. My proposed aims will help understand the mechanisms in which the causal variants and target genes drive the AD risk, and enhance our understanding of PICALM and EED in microglia and in AD.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT To address the unmet need for an effective treatment for the ultra-rare porphyria, Congenital Erythropoietic Porphyria (CEP), we propose a phase 1/2 clinical trial to be conducted by a team of porphyria experts experienced in successful clinical trials for other porphyrias. CEP is an autosomal recessive inborn error of heme biosynthesis resulting from mostly missense mutations that markedly reduce the activity of the heme biosynthetic enzyme, uroporphyrinogen synthase (UROS), resulting in the massive accumulation of the non- physiologic and phototoxic porphyrins, uroporphyrin and coproporphyrin I isomers. Pathophysiologically, the pathogenic porphyrins accumulate in erythroid cells, leading to hemolysis, with the resultant severe clinical manifestations of transfusion-dependency, splenomegaly, and phototoxic cutaneous blistering on sun-exposed skin that often become infected, causing disfiguring photomutilation. Although bone marrow/hematopoietic stem cell transplantation has been curative in transfusion-dependent children, the limitations of match, ablation damage, graft rejection, graft vs. host disease, expert availability, etc. limit its use in clinical practice. Chronic erythrocyte transfusions and iron chelation do not treat the underlying disease pathogenesis. Therefore, the recent discovery of a pharmacologic chaperone, ciclopirox, previously FDA-approved as a topical fungicide, has been modified for oral administration. This repurposed oral form, ciclopirox olamine, has been shown in in vitro studies to increase the stability and activity of most known UROS missense mutations present in over 80% of known CEP patients. In the knock-in murine CEP model, it increased residual enzymatic stability and activity, thereby reducing accumulated porphyrins, splenomegaly, and cutaneous phototoxic damage. Importantly, this oral drug would treat the pathogenesis of the disease as has been successfully accomplished with specific chaperone therapies for Cystic Fibrosis and Fabry disease. A recent toxicity study in 34 normal adults found the oral drug well tolerated. As the severity of CEP varies from hydrops fetalis to later-onset transfusion-independent patients with severe phototoxicity, a phase 1/2 “N-of-1” study is proposed to evaluate the safety and efficacy in adult patients. The proposed clinical trial involves a six-month run-in, a three-month open-label treatment period with 2mg/kg of ciclopirox olamine, followed by nine months of 4mg/kg. As proof-of-concept endpoints, patients will be evaluated to determine the drug's effect on blood hemoglobin levels and symptoms of anemia, sunlight tolerance, porphyrins, hematologic measures including percentage of fluorocytes and reticulocytes, and quality of life measures.

NIH Research Projects · FY 2025 · 2025-09

Without the occurrence of cancer cell dissemination (metastasis), most breast cancer patients would survive following diagnosis and treatment. Alas, cancer cells can disseminate to distant sites long before diagnosis of the primary tumor. Based on clinical investigations, bone is established as the most predominant site of metastasis in breast cancer. Over the years, we started to realize a preferential association of breast cancer subtypes with certain metastatic sites. For instance, hormone receptor- positive (HR+) breast cancer, which represents the most common subtype of breast cancer (65-70%), has a high propensity toward skeletal tissues compared to HR- subtypes (TNBC and HR-/Her2+). Accumulating evidence indicates a key role of bone resident cells (osteoblasts, osteoclasts, and vascular cells) in these processes. Although multiple mechanisms can contribute to these processes, the clinical implications remain limited. Recently, we identified a bone-induced factor (NRG3) that may help switch current paradigms. NRG3 is part of the neuregulin family of ligands that modulate erbb receptors. While the role of Neuregulins is still enigmatic in breast cancer, our preliminary results suggest strong implications in metastasis progression. Here, we will use molecular approaches to (i) determine and functionally characterize the NRG3 regulome induced in the bone microenvironment, (ii) identify the NRG3 signaling using genetic approaches, and (iii) assess the impact of NRG3 depletion on secondary metastasis. Considering the well-established role of Receptor Tyrosine Kinases such as Her2 in stemness, DTC, and metastasis progression, our proposed investigation may uncover novel bone-mediated properties with high therapeutic potential.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT Kinase enhancing mutations in Leucine-Rich Repeat Kinase 2 (LRRK2) are associated with both familial and sporadic forms of Parkinson’s (PD), including the prevalent G2019S mutation. In brain, LRRK2 is highly expressed in striatal projection neurons (SPNs) and cortical neurons. Our lab has previously shown significant AMPA receptor (AMPAR) trafficking deficits in mouse Lrrk2G2019S SPNs, however the molecular mechanisms underlying these trafficking deficits is unknown. LRRK2 phosphorylates a number of Rab GTPases, including Rab8, and previous studies in hippocampal neurons suggest a role for Rab8 in AMPAR trafficking. Rab8’s interactions with specialized proteins that maintain a tightly regulated activation cycle are impaired by LRRK2- mediated phosphorylation, suggesting that Rab8 activity is further regulated by LRRK2 phosphorylation. How Rab8 regulates AMPAR trafficking in SPNs, and the functional consequences of LRRK2-mediated phosphorylation of Rab8, are not known. Preliminary evidence suggests that both phosphorylated and unphosphorylated Rab8 interact with the AMPAR subunit GluA1 in vitro. Therefore, I hypothesize that Rab8 phosphorylation state regulates AMPAR trafficking in SPNs, and excess phosphorylated Rab8 (p72-Rab8; pRab8) mediates AMPAR trafficking deficits seen in the Lrrk2G2019S mutation. To address these gaps, the proposed experiments aim to characterize Rab8’s functional relationship with AMPAR trafficking pathways in SPNs and explore the effects of LRRK2-mediated phosphorylation of Rab8 by combining genetic manipulations of Rab8 with imaging, biochemical, and electrophysiological approaches.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT Regular exercise promotes health and prevents disease, and for chronic suppurative lung diseases, like cystic fibrosis (CF), exercise serves an additional therapeutic role as an independent airway clearance technique. People with CF (pwCF) generally exhibit lower levels of physical fitness as compared to their healthy peers, such that regular exercise is recommended as part of the standard treatment regimen for CF. Beyond its physiological, functional, and psychological benefits, regular exercise is thought to exert anti-inflammatory effects and reduce infection susceptibility. Understanding how the immune modulatory effects of regular exercise influence CF disease progression, along with the underlying mechanisms, will help to determine the optimal type and dose of exercise needed to improve the overall fitness and modify the clinical course of pwCF. The proposed research project aims to 1) generate hypotheses regarding the impact of regular exercise on CF disease progression and 2) identify blood biomarkers, reflecting lung-specific responses to exercise, that can be applied to the study of human health and disease. Leveraging the Molecular Transducers of Physical Activity Consortium (MoTrPAC) young adult rat endurance exercise training dataset, in conjunction with publicly available CF and healthy control datasets, we will conduct a cross-species comparison of differentially expressed gene modules in the lungs and blood of young adult rats undergoing endurance exercise training with those in pwCF. This comparison will help to generate hypotheses about the impact of regular exercise on CF disease progression, focusing on the lungs but considering the broader impact on the entire body. We will additionally use the MoTrPAC young adult rat endurance exercise training dataset to identify a panel of sexually conserved blood biomarkers associated with lung-specific responses to endurance exercise training. The successful completion of the proposed research project will generate a set of testable hypotheses regarding the impact of regular exercise on CF disease progression and a panel of blood biomarkers to facilitate the translation of the MoTrPAC young adult rat endurance exercise training data into human studies.

NIH Research Projects · FY 2025 · 2025-09

Abstract The role of exercise in improving health and fitness has become an increasingly important area of scientific research. The MoTrPAC multiomic rat endurance exercise training (EET) study has convincingly shown that the effects of exercise are pervasive throughout the body and these effects are found at all omic levels. The study measured the effects of up to eight weeks of EET on transcriptomic, epigenomic, proteomic and metabolomic features across up to 19 tissues in male and female rats. In order to fully understand the mechanisms by which exercise impacts the body and health, we need a whole body, multiomic model of the regulatory networks that drive EET-induced molecular changes. One critical aspect of gene regulation that was overlooked in the MoTrPAC rat data analyses is exercise-regulation of alternative mRNA splicing (AS). Initial studies of the acute human MoTrPAC data show dramatic regulation of AS by acute exercise in the limited tissues sampled in that study. This indicates that exercise-regulated splicing is an important mediator of the effects of exercise and motivates our proposed study of the effects of EET throughout the organism. We propose to develop a 19 tissue map of the AS landscape, both at baseline and in response to EET. This will be an unprecedented diversity of tissues with which to explore AS behavior. By integrating these results across omes, we can elucidate both what is changing in the AS landscape and how it affects downstream gene and protein activity. In addition, we can use epigenome and transcription factor activity analyses to identify the mechanisms underlying AS regulation. First, we will measure baseline un-exercised tissue specificity of AS events, and second, we will find differential AS events (DASs) in response to one, two, four, or eight weeks of EET. We will modify bioinformatic integration tools developed by our team to optimize integrated analysis of splicing and regulatory mechanisms, including PLIER to look for cross-tissue patterns of AS response to EET annotated to enriched pathways and MAGICAL to generate regulatory circuits of coordinated EET responses in chromatin accessibility or DNA methylation with DASs and enriched transcription factors. This knowledge will be a valuable resource for future AS analysis in general and, specifically, in studies of exercise. Because most of the tissues included in this study cannot be analyzed in human subjects, we will use these results as a foundation for future grant proposals seeking to connect the 19 tissue AS responses in rat to human understanding of the AS landscape.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Even with effective antiretroviral therapy (ART) that suppresses viral replication, individuals with HIV frequently experience chronic neuroinflammation and immune activation, leading to increased risk of neurocognitive disorders and other health complications. The connection between persistent HIV reservoirs in the central nervous system (CNS) and ongoing neuroinflammation remains poorly understood. This project investigates the role of the NLRP3 inflammasome in HIV-induced neuroinflammation, with a particular focus on how inflammasome activation contributes to blood-brain barrier (BBB) dysfunction and the persistence of HIV latency in the CNS. The research team, with expertise spanning HIV biology, inflammasome pathways, CNS model systems, and advanced data analysis, is uniquely equipped to address this critical gap in knowledge. Aim 1: This aim seeks to elucidate the link between HIV-1 infection and NLRP3 inflammasome activation in the CNS. We will use advanced brain organoid models and humanized mouse models to study how HIV-1 triggers inflammasome activation in microglia and contributes to BBB breakdown, providing insights into the mechanisms that sustain neuroinflammation and HIV persistence. Aim 2: This aim investigates the impact of NLRP3 inflammasome activation on the establishment and maintenance of HIV latency in the CNS. By examining how inflammasome activation influences the behavior of latent HIV reservoirs, the study will explore potential pathways that could be targeted to disrupt latency and reduce inflammation. Aim 3: This aim focuses on developing and evaluating therapeutic strategies to mitigate neuroinflammation and HIV persistence. Using innovative lineage tracing techniques and advanced in vivo models, we will test small molecule inhibitors and other therapeutic interventions to assess their efficacy in reducing inflammasome-driven inflammation and preserving CNS function. This project is expected to yield significant advancements in understanding the mechanisms driving HIV-associated neuroinflammation and latency. The findings from this project will provide crucial insights into the role of the NLRP3 inflammasome in sustaining HIV persistence, identify novel therapeutic targets, and potentially pave the way for a functional cure for HIV, ultimately leading to improved treatment strategies and quality of life for people living with HIV.

NIH Research Projects · FY 2025 · 2025-09

In Alzheimer’s disease and related disorders (ADRD) research, there is a lack of ethnic/racial diversity at every level, from investigators to regulators, monitors and clinical research coordinators and administrators. In this R25, we propose to address this issue by developing and implementing the Postbac Alzheimer’s Disease Development Initiative (PADDI), a comprehensive two-year program, to train postbacs about issues related to the design and implementation of AD/ADRC research methodologies, and the ethical conduct of research. The Mount Sinai ADRC, which has been funded since 1984, is an ideal host for this training endeavor. Based on our collective experiences, which include decades of teaching, mentoring, directing training programs, curriculum development, outreach and recruitment for a wide range of ADRD studies, and supervising trainees at all levels, we know that early engagement in science coupled with clinical experience with those with cognitive impairment leads to a motivated generation of allies, staff, clinicians and investigators with the passion and commitment to address these diseases. We will partner with the numerous training environments at Mount Sinai to provide educational opportunities for the postbacs and collaborate with several ADRC, Mount Sinai, and community outreach resources to attract young college graduates interested in a scientific career. PADDI researchers will spend 75% of their time as clinical research coordinators (CRCs) and 25% of their time in didactics and other professional development activities. The proposed program will include the design of a comprehensive recruitment, mentorship and career development strategy that targets recent college graduates from local public universities and colleges . Curriculum will encompass training in fundamentals across three areas including clinical, research and regulatory processes. PADDI will provide hands-on clinical research training and promote the development of scientific communication and leadership skills. Professional networking opportunities will be provided through collaboration with other MSHS training programs. The program will provide mentor training and evaluate their success. We will implement robust administrative, evaluation and tracking mechanisms to gauge the PADDI program’s success and incorporate feedback from our Advisory Committee. Finally, PADDI program results and materials will be disseminated online and presented at relevant meetings and workshops.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY Effective antiretroviral therapy (ART) has dramatically reduced mortality, therefore people with HIV (PWH) are now surviving into middle and old age. Despite this success, PWH experience high rates of multimorbidities, and functional and cognitive decline at a younger age than those without HIV, a phenomenon known as accelerated aging. Thus, there is an urgent need to focus not just on mortality but on healthspan, or the time someone is healthy not just alive, by defining the drivers of the accelerated aging. Epigenetic clocks are powerful tools for predicting aging-associated diseases and lifespan in the general population; however, there are little data on the effects of HIV and ART on epigenetic aging in older PWH. More recently, clonal hematopoiesis, a noncancerous expansion of a somatic blood cell clone, of indeterminate potential (CHIP) has been shown to be associated with increased risk for hematological cancers, cardiovascular (CVD) and pulmonary diseases, obesity, osteoporosis, and all-cause mortality, in the general population. Importantly, the prevalence of CHIP is much higher and onset earlier among PWH. Moreover, CHIP has been more commonly identified in PWH with CVD, sometimes years before CVD diagnosis. However, data are lacking on its contribution to other age-related outcomes in PWH. Moreover, the modifying effects of underlying genetic predisposition, socio-behavioral aspects and other HIV- specific risk factors are largely unknown limiting our ability to prevent or slow down functional and cognitive decline and multimorbidity. CFAR Network of Integrated Clinical Systems (CNICS) is a large, well-characterized, prospective gender, and racially/ethnically diverse cohort with ~49,000 PWH. CNICS captures rich longitudinal clinical data, patient-reported measures and outcomes, health-related quality of life (HRQL), and additional social determinants of health factors, and provides access to a biospecimen repository. We will leverage and build on the extensive platform and data collection of CNICS to identify the drivers of accelerated aging. We hypothesize that the interplay of epigenetic changes and CHIP with genetic predisposition and HIV-specific and socio- behavioral factors drive functional and cognitive decline, multimorbidity, and mortality among PWH. In Aim 1, we will establish a sub-cohort from 6,442 already enrolled PWH ages ≥50 years with existing biospecimens and prospective clinical data and determine the role of the epigenetic aging, genetic predisposition, and socio- behavioral factors in functional and cognitive decline, osteoporosis, multimorbidity, HRQL, and morbidity in PWH. In Aim 2, we will generate whole exome sequencing data to identify specific CHIP-driving mutational patterns that define different risk of developing adverse age-related outcomes and explore the interplay between CHIP and epigenetic changes in PWH. In Aim 3, we will use Mendelian randomization to detect the causal effect of epigenetic aging and CHIP on age-related diseases and identify potential drugs that may reverse adverse outcomes. Understanding the predisposition to and the impact of accelerated aging is critical to identifying PWH at high risk, developing preventive and therapeutic strategies, and improving the healthspan in the setting of HIV.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Approximately 150,000 vertical HIV transmission events occur annually via breastfeeding; yet, only ~10-15% of infants breastfed by a mother living with HIV will become infected, suggesting components in human milk are partially protective against HIV infection [1, 2]. Human milk is comprised of cells that are >90% viable and remain functional well beyond ingestion, with infants likely to ingest ∼105–108 milk leukocytes daily [3, 4]. Functionality is often in conjunction with milk antibodies (Abs). The level and type of HIV-specific Abs in milk have been correlated with reduced milk vertical transmission (MVT), but what remains largely unexplored is the contribution of the cellular fraction [5]. Abs elicited against HIV need not be neutralizing to exhibit anti-viral effects, and numerous studies have demonstrated Fc-mediated Ab effector functions such as Ab-dependent cellular phagocytosis (ADCP) and Ab-dependent cellular cytotoxicity (ADCC) to be critical in protecting against HIV [6-10]. Our studies using milk obtained from HIV- women in the USA demonstrated milk leukocytes can perform ADCP of HIV targets when elicited by any IgG isotype, mediated by granulocytes as well as various myeloid cell types [11]. This work identified a significant population of atypical CD14- myeloid cells in milk, many of which appeared capable of ADCP. These data call for a comprehensive analysis of milk myeloid cells, especially in the context of their role in MVT of HIV – both as vehicles of and defenders against transmission. Not only must these cells be classified and compared across lactation periods, but their anti-viral functionality and HIV susceptibility determined as well [12]. Herein we intend to analyze milk obtained from donors recruited as part of the UPenn-Botswana Partnership (BUP), a subsidiary of the University of Pennsylvania, and therefore living in a geographic context relevant for MVT. We will compare milk cells from HIV- and HIV+ participants, including a subset of HIV+ participants on antiretroviral therapy (ART) with suppressed viral loads. The proposed project aims to test the hypothesis that transcriptional profiling will reclassify and reveal novel milk myeloid cell subsets unique from those expected in PBMCs, and that certain milk myeloid subsets will be susceptible to HIV infection and/or exhibit Fc-mediated function, thus identifying cellular targets to eliminate or to augment as part of a precision therapeutics strategy for lactating HIV+ women designed to eradicate MVT. As such, we aim to: 1) Conduct an in-depth analysis of human milk myeloid cells obtained from HIV- and HIV+ donors using single cell RNA sequencing (scRNAseq) in order to define cell subsets based on transcriptional profiling; 2) Determine the HIV susceptibility and HIV Envelope expression profile of each newly-defined milk myeloid subset; and 3) Assess the Fc-mediated functionality of each newly-defined milk myeloid subset. The overall goal of this proposed foundational work is to ultimately develop a targeted, therapeutic vaccine for breastfeeding women living with HIV to eliminate MVT. This work is focused on the unique needs of these women and their infants, who are doubly underserved being in a low-income country setting.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Maternal depression affects approximately one in six new mothers worldwide. Left untreated, there is an increased risk of adverse child development, self-harm, partner/marriage dissolution, and self-medicating, which can lead to disastrous effects on the mother, child, and family. Despite its prevalence, the multifactorial etiology of maternal depression and the underlying biology is poorly understood. Emerging evidence indicates that stress (i.e., social exposome) and the built environment (i.e., urban exposome) may be causal factors in depression through affecting key biological pathways such as inflammation, hormones and steroids, and tryptophan metabolism. Indeed, our preliminary data links PM2.5 exposure (i.e., particulate matter with diameter <2.5 μm) with maternal depression and demonstrates that psychosocial stress during pregnancy exacerbates maternal depression. However, a study of the complex interactions between the multi-level, built environment and psychosocial factors that describe urban living during pregnancy—built environment (i.e., PM2.5, NO2, noise, light-at-night, greenness, temperature, noise, and population density) and multiple stress constructs (i.e., negative life events, salivary cortisol, and coping mechanism)—have not been robustly investigated. We will address this gap using rigorous epidemiologic and statistical methods. We will leverage the Programming Research in Obesity, GRowth, Environment and Social Stressors (PROGRESS) longitudinal birth cohort, which includes a comprehensive questionnaire and biological measures of psychosocial stress, high temporally- and geo-spatially resolved built environment measures, and longitudinal maternal depression phenotyping and biospecimen collection from pregnancy until 14 years postpartum. To address multiple mental health constructs, we will characterize maternal depression across this critical decade of maternal life using a validated questionnaire (longitudinal EPDS collected 12 times) and the gold standard in clinical depression diagnosis (SCID-5). We will use state-of-the-art satellite remote and ground sensing models to investigate the impact of the urban exposome during pregnancy on the risk of maternal depression (postpartum, depression progression over 14 years, and clinical depression up to 14 years post pregnancy). We will then determine the role of the social exposome alone and in combination with the urban exposome on maternal depression risk. We will use our metabolomics platform to simultaneously measure key neuro-immuno-endocrine metabolites in a single assay to identify metabolic changes that link the urban–social exposome with depression. Our population of 600 mothers in Mexico City with high but variable exposures to the urban–social exposome provides a unique opportunity to investigate the complexity of modifiable risk factors of urban life and early biomarkers of maternal depression. This will significantly enhance our knowledge of when, where, and how to intervene to reduce the risk and incidence of depression.

NIH Research Projects · FY 2025 · 2025-09

Depressive disorders (DD) are highly prevalent among older adults with asthma and are associated with increased asthma morbidity. Asthma affects 8% of the United States population ≥ 60 years and older adults are more likely to die from asthma and have hospitalizations. Self-management behaviors (SMB) are key for achieving good asthma control. Unfortunately, depression is a major contributor to low adherence to controller medications and other SMB. We developed Supporting Asthma Self-Management Behaviors in Older Adults (SAMBA), a self-management support (SMS) program that improves asthma outcomes. However, it was not designed for patients with DD and secondary analyses showed SAMBA was less effective in these patients. Cognitive behavioral therapy for adherence and depression (CBT-AD) was designed for patients with chronic disease and depression and improves SMB, disease control, and depression. This proposal brings together a multidisciplinary team of investigators to test the feasibility and preliminary efficacy of an integrated intervention of SAMBA and CBT-AD. The Specific Aims are to: 1) Develop an integrated 10-12 session asthma SMS program with CBT-AD for older adults with uncontrolled asthma and comorbid DD (current major depressive disorder or persistent depressive disorder) with input from patients, caregivers and other stakeholders to validate content and protocols; 2) Conduct a pilot RCT to determine the feasibility and preliminary impact on asthma (medication use tracked by electronic devices, asthma control, quality of life, and lung function) and depression (Geriatric Depression Scale scores) outcomes of the SAMBA-CBT-AD intervention compared with SAMBA alone for urban older adults with uncontrolled asthma and a current DD. An exploratory aim will assess the mechanisms (via health beliefs and asthma management self-efficacy) underlying the effect of the integrated SAMBA-CBT-AD intervention. The pilot RCT will enroll 60 older adults (≥60 years) from practices in East Harlem, New York City. Participants will undergo a baseline evaluation of their asthma and the gold standard psychiatric interview to assess DD. Outcome measures will be assessed at intervention completion (month 3), and for sustainability at 6 months. This innovative theory-based intervention addresses a significant public health problem among older patients, who are rarely the focus of interventions, to improve both depression and asthma outcomes. The project will prepare the team for conducting a subsequent larger efficacy trial of the integrated SAMBA-CBT-AD intervention.

NIH Research Projects · FY 2025 · 2025-09

Abstract H5N1 Highly Pathogenic Avian Influenza virus (HPAIV) spreads globally through migratory birds. Outbreaks of avian influenza cause significant financial losses for particularly for the poultry and dairy industries. The first human infection with HPAIV H5N1 was in 1997, followed by an additional 17 cases and a total of 6 deaths. Since 2003, there have been over 900 human cases with a mortality rate exceeding 50 percent. Currently, no effective vaccine is available, and Tamiflu does not work well against clade 2.3.4.4b H5N1. It is pivotal to understand the risk of the virus acquiring efficient cross-species transmission and further sustained human-to-human transmissibility. Adaptive mutations have emerged during infection of humans and a broad spectrum of mammals, posing a huge threat to public health. As a host restriction factor, myxovirus resistance 1 (Mx1) protein is recognized as a crucial gatekeeper candidate. Studies have shown Mx1 protein exhibits antiviral activity against HPAIV H5N1. The proposed research will test the hypothesis that different H5N1 isolates have different sensitivity to different Mx1s, which is associated with mutations of the viral genome. Preliminary data showed that murine Mx1 (muMx1) provides impressive resistance against H5N1 infection. Mx1 expression increased the lethal dose by at least 1000-fold when comparing that between wild type B6 and Mx1 (+/+) A2G mice. Meanwhile, the Mx1 sensitivity differs significantly between a human isolate and a sea lion isolate, carrying limited variation between two viral genomes. I will test the sensitivity of different H5N1 viruses to muMx1 in vivo and investigate any correlation to specific genotypic polymorphisms (Aim 1). It is anticipated that the data will show different H5N1s carry different muMx1 sensitivity. Based on the genome alignment, the genomic determinant will be identified and related to Mx1 sensitivity. I will investigate the impact of these new biomarkers on virus sensitivity to Mx1. The results will provide insight into the risk of H5N1 breaking the species barrier with novel genetic markers. Additionally, I will assess the sensitivity of H5N1 viruses to Mx1 from different hosts in vitro, serving as a tool to assess potential host range (Aim 2). A549 STAT1 knockout cell line will be used as a platform to express exogeneous Mx1 proteins to examine Mx-mediated antiviral phenotype. It is anticipated that the results will show viruses from different hosts display different Mx1 sensitivities, indicating each virus has its own susceptible hosts. This Mx1 library can be expanded with new Mx sequences from mammalian species of interest, assessing the potential transmissibility of certain H5N1s among different mammalian species. The overall project will provide risk assessment of the current HPAIV H5N1 to public health and in vitro and in vivo models for the future study. Completion of this proposal will provide training in (1) advanced molecular biology approaches, (2) virology concepts and methods, (3) next generation sequencing skills, (4) mass spectrometry techniques, and (4) biosafety procedures for working with select agents – equipping me with essential expertise needed to develop an independent research program on emerging infectious diseases.

NIH Research Projects · FY 2025 · 2025-09

Project Summary / Abstract Coronary artery bypass grafting (CABG) is the most common adult cardiac surgery procedure, and the standard of care for patients with severe coronary artery disease. CABG can be performed using either arterial or venous grafts, with arterial grafts being associated with better outcomes in observational studies, but not in the only published randomized trial. Women represent a minority of patients in CABG trials, and there are important biological and surgical differences between men and women undergoing CABG ROMA:Women is the first trial comparing arterial vs. venous graft in a exclusive women population. In a rigorous randomized trial that focus enrollment exclusively in women we will determine 1) the impact of MAG vs SAG on major adverse cardiac and cerebrovascular events in women undergoing CABG and 2) the impact of MAG vs SAG on generic and disease- specific quality of life (QOL) as well as physical and mental health symptoms in women undergoing CABG. We will enroll 2,000 women randomized 1:1 to MAG or SAG. ROMA:Women will leverage the existing infrastructure of ROMA (conducted on both Women and Men) including the clinical trial database, case report forms, randomization system, site training resources, regulatory approvals, network of participating sites, and study coordinators. The trial primary outcome will be a composite of death, stroke, non-procedural myocardial infarction, repeat revascularization and hospital readmission for acute coronary syndrome or heart failure at a minimum follow-up of 2.5 years. The secondary outcome will be the absolute changes in the Seattle Angina Questionnaire (SAQ) at 12 months compared to baseline. The trial is designed to have >90% power to demonstrate a 25% relative risk reduction in the primary composite outcome in the MAG group. Generic QOL (SF-12, EQ-5D), and symptoms (PROMIS-29) as well as key clinical events will also be captured.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Applications for messenger RNA (mRNA) in a wide range of clinical indications—such as pathogen vaccination, cancer immunotherapy, protein replacement, and gene editing—are rapidly advancing. Since Moderna and Pfizer’s mRNA vaccines against SARS-CoV-2 highlighted the efficacy and potency of mRNA technology, there has been ongoing effort to optimize the formulation of the mRNA and the delivery vehicle, which is often a lipid nanoparticle (LNP), for novel applications. One of the major challenges faced by mRNA therapeutics, even those that have targeting moieties, is mRNA uptake and expression by off-target cells. Current mRNA vaccines depend on antigen presentation by antigen-presenting cells (APCs) to induce robust immunity, yet hepatocytes and skeletal myocytes are also transfected, leading to unintended immune modulation. To better understand how these two cell types influence adaptive immunity to the mRNA-encoded protein, I will regulate the cellular expression patterns of delivered mRNA by incorporating target sites complementary to cell-type specific, endogenous miRNA. My preliminary data indicate that hepatocytes dampen the magnitude of CD8+ T cell responses to the mRNA-encoded antigen, whereas skeletal myocytes enhance CD8+ T cell responses. Understanding the mechanisms underlying the immunomodulatory effects of these two cell types is crucial for optimizing mRNA-based therapeutics. My central hypothesis is that hepatocytes drive an immunoregulatory response to mRNA-encoded protein through abortive CD8+ T cell stimulation and regulatory T cell (Treg) induction, while skeletal myocytes can directly present antigen to and activate CD8+ T cells. I will engineer clinically-relevant mRNA formulations with regulated mRNA expression in these two cell types to enhance antigen-specific adaptive immunity while reducing deleterious off-target effects. In Aim 1, I will delve into the mechanisms by which hepatocytes and skeletal myocytes modulate antigen-specific T cell immunity against the mRNA-encoded protein. I will specifically evaluate the effect of Treg depletion and hepatocyte-restricted antigen presentation on CD8+ T cell responses. To assess the role skeletal myocytes may play, I will evaluate the duration of antigen presentation and whether skeletal myocytes are sufficient to induce T cell recall. I will also investigate whether exogenous antigen uptake by APCs can induce antigen-specific T cell immunity. In Aim 2, I will leverage the impact of mRNA expression by hepatocytes and skeletal myocytes to enhance antigen-specific adaptive immune responses to tumor-associated antigen (TAA) and SARS-CoV-2 spike mRNA vaccines, while reducing off-target cytotoxicity from CD8+ T cells, especially in the liver. I will design mRNA vaccines to boost anti-TAA CAR-T cells, induce exogenous anti-tumor CD8+ T cell activity, and enhance anti-spike antibody and T cell responses. Ultimately, this project aims to provide insight into design strategies for mRNA vaccines to tailor immunogenicity while minimizing adverse effects.

NIH Research Projects · FY 2025 · 2025-09

Chinese Americans are one of the fastest-growing populations in the U.S.; however, their participation in Alzheimer’s Disease and Alzheimer’s Disease-Related Dementias (AD/ADRD) research remains low. This lack of representation may mask health outcomes in this group, which varies by generation status (foreign- vs. native-born) and social determinants of health (SDOH). SDOH factors include, but are not limited to acculturation, language proficiency, healthcare access, educational quality, stress, environmental exposures, and social support networks, which can contribute to health disparities in Chinese American older adults, worsening AD/ADRD outcomes. However, the relationship between SDOH and AD/ADRD risks in this population is not well-studied, particularly in relation to other health issues like sleep disruption, which is prevalent in older populations. Consistent with the existing literature, our preliminary research suggests that foreign-born Chinese Americans experience poorer sleep quality. However, it is unclear whether there is an association between sleep disruption and elevated AD/ADRD biomarkers, as has been observed in non-Hispanic White participants. Overall, differences in cognitive function and AD/ADRD biomarkers may exist between foreign- and native-born older Chinese Americans due to differential experiences of SDOH, including stress, which has been linked to AD/ADRD-related functional/structural changes (e.g., hippocampal atrophy) and increased levels of Aβ production in other groups. Our team demonstrated that increased levels of distress was associated with higher levels of plasma p-tau level and better performance on tests of attention, executive function, processing speed, and memory in a small sample of older Chinese Americans. Yet, a limited number of studies have explored these relationships specifically in a well-characterized sample of Chinese American older adults born in and outside of the US. This study aims to address these gaps by examining the interplay between SDOH, sleep disruption, and biological factors in AD/ADRD risks among older Chinese Americans. The research team will recruit 250 participants (125 foreign-born and 125 native-born) for baseline evaluations and 200 participants (100 foreign- born and 100 native-born) for one- and two-year follow-ups. The study will use objective measures like at- home sleep tests, cognitive testing, PET/MR, and blood-based proteomic and single cell multiomic biomarkers to investigate how sleep disturbances and SDOH contribute to AD/ADRD risks. Additionally, a novel, integrative network biology and machine learning (ML) based approach will be employed to develop highly predictive diagnostic and prognostic biomarkers of AD/ADRD by integrating all clinical, imaging and multiomic data. This project, consistent with NIH priorities, will provide crucial data to understand the unique health disparities faced by Chinese Americans in AD/ADRD research.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Most influenza cases resolve without complications, but some progress to serious conditions like pneumonia, acute respiratory distress syndrome, or encephalitis. Genetic predispositions, such as inborn errors of immunity, are known factors that contribute to these severe outcomes, yet the underlying mechanisms in otherwise healthy individuals, particularly children, remain unclear. This proposal focuses on ANKEF1, a ciliary protein identified as having a loss-of-function mutation in a child who succumbed to influenza-associated rhombencephalitis—a rare and severe complication characterized by inflammation of the brainstem that can occur in some severe cases of influenza-associated encephalitis (IAE). Our preliminary findings indicate that ANKEF1 is critical for ciliary motility in airway epithelial cells, which is essential for effective mucociliary clearance (MCC). In mouse models, ANKEF1 deficiency leads to impaired MCC, reducing the ability to clear pathogens from the respiratory tract and leading to increased susceptibility to influenza infection, higher morbidity, and greater mortality. We hypothesize that persistent viral presence in the airways due to defective MCC triggers an exaggerated inflammatory response, which elevates the risk of IAE through increased cytokine production and disruption of brain barrier integrity In this proposal, we will leverage novel mouse models, human iPSC-derived airway epithelial cultures, and advanced multi-omics technologies to define how ANKEF1 regulates ciliary function and host defense. Aim 1 will investigate how ANKEF1 deficiency alters ciliary beat frequency, coordination, and motility in the airways and brain, and assess the resulting impact on viral replication and immune responses ex vivo. Aim 2 will characterize disease progression in ANKEF1-deficient mice by measuring viral titers, immune responses, and tissue damage in both the respiratory system and brain. Additionally, we will use single-cell RNA sequencing and spatial transcriptomics to identify key pathways associated with increased disease severity. Aim 3 will elucidate the molecular mechanisms by which ANKEF1 modulates ciliary functions and identify key interactions and regulatory networks that contribute to host defense against IAV infection. This research will provide new insights into the role of ciliary function in airway and brain defenses, potentially revealing therapeutic targets for severe influenza and other respiratory diseases.

NIH Research Projects · FY 2025 · 2025-09

I am an environmental epidemiologist with a main research focus on the “exposome” and its role in obesity-related outcomes and chronic disease. The training presented in this proposal will enable me to become an independent public health researcher in the field of environmental epidemiology, specializing in endocrine-disrupting chemical (EDC) mixtures and long-term cardiometabolic health in women. This project addresses the role of EDCs in the onset of metabolic syndrome (MetS), which is a cluster of conditions that can increase the risk for heart disease and type 2 diabetes, as well as examines potential underlying mechanisms. This opportunity will allow me to hone the skills needed to become an independent investigator by 1) expanding my knowledge in exposure-mixture methods, 2) acquiring the tools to analyze and interpret high-dimensional metabolomics data, and 3) advancing my expertise in a relatively highly prevalent, yet understudied, metabolic condition. The rising MetS epidemic has reached alarming rates in North America, with a prevalence in adults of more than one-third in the US. Currently, the etiology of MetS and its mechanisms of action are poorly understood. An emerging risk factor for MetS is exposure to environmental “obesogens”, such as EDCs. Endocrine disruptors can act in the body by mimicking steroid hormones and increasing oxidative stress, thereby triggering MetS components such as hypertension, abdominal adiposity, hyperglycemia, or dyslipidemia. Moreover, pregnancy is likely a susceptible period that could exacerbate the deleterious effects of EDCs with long-lasting consequences for women’s health. I will leverage ~400 pregnant women from the Programming Research in Obesity, Growth, Environment, and Social Stressors (PROGRESS) prospective cohort to examine 1) the longitudinal association between EDC exposure-mixtures—phthalates, metals, air pollutants, pesticides, and PFAS—and MetS onset, 2) the subsequent alteration in individual metabolites and pathways stemming from these exposures, and 3) the mediating metabolomic features characteristic of MetS. The training originating from the K99 phase of this proposal will ensure the correct implementation of chemical mixtures-MetS association analyses in the first phase of the award (K99), and of mixture- omics and mediation analyses during the second phase (R00). A renowned multi- disciplinary team of experts will be convened to help me achieve my goals, with whom I will meet and discuss rigorous training related to environmental endocrinology and metabolism (Dr. Damaskini Valvi), interpretation in longitudinal cohort data (Dr. Martha Téllez-Rojo), maternal health (Dr. Lida Chatzi), metabolomics (Dr. Dinesh Barupal), and statistical methodology for exposure-mixtures and mediation analysis (Dr. Elena Colicino). Formal coursework, scholarly seminars, guided readings, conferences, and workshops will also be incorporated into my curriculum, collectively helping me build my career as an independent environmental epidemiologist with a focus on cardiometabolic diseases.

NIH Research Projects · FY 2025 · 2025-09

Summary Skin diseases exhibit varying prevalence, severity, and responses across groups of different ancestry, yet the underlying reasons remain unclear. A recent study highlighted that hundreds of genes are uniquely expressed in the skin of healthy individuals of African American ancestry compared to those of European, indicating the profound diversity within the human species. DNA regulatory elements, including promoters and enhancers, are crucial in controlling gene expression by acting as switches to turn genes on or off. However, most studies on transcriptional control of epidermal keratinocytes have been done in European ancestry populations, thus limiting the utility of the results across a variety of human populations. This R21 application is submitted in response to NOT-AR-24-009: Accelerating Research in Understudied Skin Types and is focused on the special interest research topic “using functional genomics, epigenomics, and transcriptomics to identify lead candidate genes and transcripts associated with increased prevalence and severity of skin diseases in patients with understudied skin types.” In response to the NOT-AR-24-009, we will test the hypothesis that the observed differences in gene expression in skin cells from different ancestral backgrounds are due to the variation in DNA regulatory elements. We assembled a multi-disciplinary team of clinicians, biostatisticians, and bioinformaticians to test this hypothesis. Our team has access to human skins collected from healthy individuals of different ancestry. We developed a flow-cytometry-based strategy to isolate epidermal keratinocytes from human skin. We also refined the ATAC-seq and CUT&RUN assay using isolated epidermal cells to identify DNA regulatory elements. Armed with these technological advancements, we will identify common and unique DNA regulatory elements in epidermal cells obtained from individuals of different ancestry. By integrating our findings with existing GWAS data, our goal is to pinpoint SNPs within these regulatory elements associated with atopic dermatitis, a chronic skin inflammatory condition with a higher severity among individuals of African American and Asian ancestry. We will then functionally validate the identified DNA regulatory elements and SNPs located in these regions on controlling gene expression by performing the Massively Parallel Reporter Assay. Our focus on healthy skin samples is intended to establish a fundamental understanding of transcriptional and epigenetic differences in epidermal keratinocytes in individuals of understudied skin types, which is one of the priorities of NOT-AR-24-009. Uncovering these differences will provide much-needed information on differences in gene expression observed in different skin types and can contribute to understanding the different presentations of various skin diseases in groups of different ancestry.

NIH Research Projects · FY 2025 · 2025-09

Our long-term goal is to understand the contribution of stress-induced neural signals to glucose regulation and the development of metabolic diseases with the aim of developing better therapies. To achieve this goal, we need a more detailed understanding of the neural circuits modulated by internal and external signals that, in turn, regulate metabolic function. In this proposal, wewill create a detailedand systematic analysis of the activity of specific amygdala neural populations in response to stress, determine their physiological roles, identify genetic markers for stress-responsive neural populations with defined projections and assess the contribution of these populations to metabolic regulation in acute and repeated stress. Our pilot data strongly support the activation of specific populations of amygdala neurons in response to stressors and chemogenetic activation of these neurons and circuits rapidly and robustly increases blood glucose. Our proposal will create new insights into the neural populations and circuits that regulate metabolism bring an unprecedented level of precision and specificity by combining cell-type specific neural tracing, spatial transcriptomics and targeted neuromodulation in which we have extensive experience. Without this detailed knowledge, our understanding the contribution of defined amygdala circuits to metabolic responses to stress will remain incomplete. The overall objective of this proposal is to understand the structure, genetic composition and function of defined amygdala circuits. Our central hypothesis is that defined medial amygdala neural populations and their hypothalamic projections are critical to the metabolic responses to stress. The rationale that underlies the proposed research is that internal and external stressors result in profound metabolic responses. However, we do not know the activity or genetic identity of the activated neural populations, their downstream circuits, or physiological roles. These represent major gaps in our understanding. To test our central hypothesis and attain the overall objective, we will a) determine the activity and physiological roles of defined medial amygdala neurons in the glucose response to acute and repeated stress and b) identify the neural activity, function and gene markers for specific downstream circuits that regulate blood glucose in responses to acute and repeated stress. To do so, we will use in vivo calcium imaging to assess neural activity, cell type and circuit- specific neuromodulation to define physiological roles of defined amygdala circuits in the regulation of glucose with stress, and spatial transcriptomics to identify gene markers for neural populations with defined projections contributing to glucose regulation in stress. The proposed studies will provide a comprehensive understanding of the unique biology of medial amygdala circuits regulating metabolic function in response to acute and repeated stress to form a crucial foundation for future studies to identifying potential therapies for stress-associated metabolic diseases such as type 2 diabetes. in response to homeostatic challenges. We will

NIH Research Projects · FY 2025 · 2025-09

Project Summary. Accumulating evidence suggests that Alzheimer’s disease (AD) has a long preclinical phase, lasting years to decades during which the accumulation of pathological proteins such as amyloid beta (Aβ) and tau can occur prior to the appearance of overt cognitive symptoms. Obstructive sleep apnea (OSA) is one of the most common sleep disorders and is composed of both sleep disruption (SD) and intermittent hypoxia (IH). Disentangling the separate contributions of chronic SD and chronic IH may reveal how OSA impacts risk for AD. While the effects of SD and IH on Aβ have been studied in both humans and animal models, less is known about the effects of SD and IH on tau spread and propagation throughout the brain, a crucial step in the formation of neurofibrillary tangles (NFT) throughout the brain. Our rationale for targeting tau stems from our own findings demonstrating significant associations between cerebrospinal fluid (CSF) concentrations of the wake-promoting neuropeptide orexin and both total and hyperphosphorylated tau, and that individuals with OSA might show signs of MCI and AD at a younger age. In addition, the treatment of OSA has been shown to delay the age of onset of MCI and to improve cognitive function in AD. Although these observations and others implicate OSA in the regulation of tau in the brain they do not establish directionality. A causal role for OSA in accelerating NFT formation would be strengthened by both chronic SD and chronic IH in mouse models of tauopathy and forms the basis for Aim 1. Given the association between tau and orexin, effective treatment of OSA holds particular promise in slowing the spread of tau throughout the brain. While hyperphosphorylation of tau is important in the formation of NFT’s, the development of AD may be separately accelerated by the spread of tau from neuron to neuron by particular aggregate conformations, a phenomenon that depends on activity of the seeding neuron. The locus coeruleus (LC) is a key brain region that expresses some of the earliest tau pathology, and importantly, the LC promotes wakefulness through rostral noradrenergic projections and is silent during sleep. Therefore, the increased neural activity of the LC during SD or IH represents a potential mechanism by which chronic SD affects the spread of tau pathology to rostral targets. Identifying the precise brain targets to which tau spreads from the LC and the time course over which this occurs is addressed in Aim 2. Addressing the hyperphosphorylation and spread of tau in models with and without LC specificity and the associated behavioral consequences on spatial, motor and fear learning as a function of chronic sleep disruption or chronic intermittent hypoxia serves as the basis for Aims 3.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Immune cells are an integral part of the adipose microenvironment and have pronounced effects on the expansion and activation of thermogenic adipocytes, also known as beige/brite adipocytes. Anti-inflammatory factors are postulated to be beneficial for insulin resistance and diabetes. Our lab recently reported that ablation of interleukin-10 (IL10)-associated anti-inflammatory signaling in mice, surprisingly, improved insulin sensitivity and glucose tolerance, protected against diet-induced obesity, and elicited browning of white adipose tissue. My proposal builds upon comprehensive data to address the crosstalk between immune cells and adipocytes within adipose tissue depots in regulating adipose thermogenesis. Our published and pilot studies show that interfering with IL10 activity in adipocytes is associated with increased thermogenesis, mitochondrial respiration, and energy expenditure. Ablation of adipose T cells increased adipose thermogenesis, pointing to a hematopoietic origin of the IL-10 signal regulating adipose tissue function. The IL10 receptor alpha (IL10R) is highly expressed in mature adipocytes, and adipocyte-specific knockdown of IL10R decreases fat mass and increases adipose thermogenesis, energy expenditure, and lipolysis. Conversely, IL10 treatment directly antagonizes the expression of thermogenic and lipolytic genes in a cell-autonomous manner. Furthermore, genome-wide Assay for Transposase-Accessible Chromatin (ATAC)-seq, ChIP-seq, and RNA-seq demonstrated that IL-10 represses the transcription of thermogenic genes by altering chromatin configuration at key enhancer and promoter regions. Our findings identify the “IL10-IL10R axis” as a novel regulator of the thermogenic transcriptional program in adipose tissue and challenge conventional assumptions regarding links between immune and inflammatory signaling and adipose tissue function in obesity. To further test the hypothesis that adipose-specific IL10R directly “senses” IL10 in the microenvironment to limit adipose thermogenesis and promote obesity-related traits I propose three Specific Aims: i) to define the therapeutic potential of ablating IL10R in the adipocytes of adult obese mice; ii) to identify IL10 secreting immune cells, and iii) to determine the mechanism by which IL10 controls adipocyte thermogenic program. Immune cells are an integral part of the adipose microenvironment; thus, studying how IL10 modulates adipocyte regulatory processes should advance our knowledge beyond inflammation and anti-inflammation and inform future therapy for obesity and insulin resistance.

NIH Research Projects · FY 2025 · 2025-09

Secretory Immunoglobulin A (sIgA) is the dominant immunoglobulin (Ig) in human mucosae and their fluids and is critical in protecting these surfaces from infection [1, 2]. sIgA is most often comprised of dimeric IgA linked by joining (j) chain. This dimer is secreted by the B cell, with j-chain marking it for transport by polymeric Ig receptor (pIgR) on epithelial cells into mucosal fluids [3]. pIgR is cleaved as it transports, leaving a domain known as secretory component (SC) attached, creating sIgA [4, 5]. SC renders it highly resistant to degradation in harsh mucosal environments [6, 7]. Human milk Ig is ~90% sIgA [8-10]. B cells that produce human milk sIgA originate from the gut associated lymphoid tissue (GALT), via the entero-mammary pathway [5, 11-13]. This pathway is evident in humans in that mammary B cells display adhesion receptors similar to GALT B cells, and milk sIgA exhibits strong reactivity for enteric microbes [14-20]. Our work analyzing the milk antibody response to SARS-CoV-2 infection found robust sIgA is elicited that is exceptionally long-lasting, a critical finding that is likely representative of any mucosal infection where immunogens ultimately present in the GALT [21-23]. In contrast, after COVID-19 vaccination, the response is IgG-dominant with low or undetectable sIgA, all waning within 3-6 months. We have also shown that influenza-specific milk sIgA is poorly boosted after seasonal vaccination [24]. Certainly, intramuscular (IM) vaccines do not elicit significant milk sIgA [24, 25]. Intranasal live-attenuated influenza vaccine is not better than IM vaccine for eliciting milk sIgA or IgG [26]. There is a true need to design vaccines aimed to elicit potent, specific sIgA responses in milk to protect breastfed infants. We expect such vaccines would be best designed to target the GALT of a lactating woman. However, animal models for such targeted vaccine design do not exist. The entero-mammary pathway is not universal; for example, little evidence has been found for this pathway in sheep or cows, who’s milk contains mainly serum IgG [27]. Early studies in mice and pigs suggested IgA-secreting GALT B cells traffic to the mammary gland, and in these species, sIgA predominates in milk [27]. Mice are not a good model animal for our work, given the paucity of milk produced and, unlike humans, their milk IgG is actively transported from the infant gut into the systemic circulation during the neonatal period [6, 28]. Pigs are likely more ideal. As such, this exploratory project aims to verify that pigs are an appropriate pre-clinical model for the human entero- mammary pathway and therefore, our future vaccine work. Using state-of-the-art single cell RNA sequencing, we will determine if, compared to other immune compartments, the cellular adhesion marker profiles of GALT and mammary gland (and/or milk) B cells destined to produce/producing sIgA are most similar; and the expressed Ig of these B cells are also highly related. The overall goal of this foundational work is to use pigs as a model for the development of targeted vaccines for lactating women that elicit robust, long-lasting milk sIgA to protect the breastfed infant. This work is focused on the unique needs of lactating women.