Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2026 · 2023-05

There is a fundamental gap in understanding how the diversity of cortical cell types and connectivity patterns translates into functional dynamics of the circuits to support cognitive behaviors. This knowledge gap hampers our understanding of the dysfunctions of decision making and other debilitating cognitive abnormalities associated with most psychiatric illnesses, including addiction, major depression, and eating disorders. My long- term goal is to unravel the intricate link from genes to circuits and to systems and reveal the pathology, pathophysiology, and behavioral deficits involved in mental disorders at the level of specific circuits and their cellular constituents. This proposal aims to determine how the genome instructs the organization and function of the premotor cortex to support decision making. The premotor cortex in mice resembles those of the non-human primates and humans, illustrating their evolutionarily conserved role in higher-level cognitive functions. In addition, we have developed behavior paradigms in mice to permit the dissection of neural circuits underlying complex behaviors using the powerful molecular tools unavailable in many other species. The central hypothesis is that molecular signatures and connectivity patterns collectively drive premotor cortex neurons to acquire distinct functions to support decision making. This hypothesis has been formulated based on previous work and the preliminary data produced by the applicants. The rationale for the proposed research is that this study will provide a new target brain area together with specific cell types and pathways for understanding and treating the cognitive deficits implicated in psychiatric illnesses. This hypothesis will be tested by pursuing two specific aims: 1) Determine the function of the molecular cell types of the premotor cortex in decision making; and 2) Establish the functional role of the afferent inputs of the premotor cortex. Under the first aim, the neural responses of individual neurons will be mapped to their molecular identity by coupling in vivo imaging and spatial transcriptomics. Further, the molecular identity will be manipulated to determine their causal contribution to function. Next, the molecular identity and function of premotor cortex neurons defined by specific afferent inputs will be established by single-cell RNA sequencing and imaging during decision making. The functional role of these afferent inputs will be further characterized by pathway-specific optogenetic manipulations. This approach is innovative because it combines in vivo imaging with spatial transcriptomics and utilizes transplantation methods and the latest circuit mapping tools to reveal the novel, cognitive role of the premotor circuit in decision making. This proposed research is significant because it answers the long-standing question about the structure and function of cortical circuits: How do neurons of distinct identities connect and interact to produce network dynamics underlying higher-level cognition. Ultimately, such knowledge has the potential to reveal the specific cell types and brain pathways underlying decision making and to better understand, intervene, and treat dysfunctions of decision making that are prevalent in psychiatric illnesses.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder with a complex genetic architecture. The development of effective therapeutics and diagnostic tools for ASD has been hindered by our incomplete understanding of underlying genetic variation. De novo variants (DNVs), estimated to contribute to 30-40% of cases, have been primarily studied in protein-coding regions of the genome. Hundreds of thousands of non-coding variants have been identified but deciphering their functional contribution to ASD etiology remains challenging. Cis-regulatory elements such as promoters and enhancers represent one avenue to assay the potential impact of non-coding DNVs, but their regulatory activity is dependent on cellular contexts such as cell type and activation state. The two cell types primarily involved in ASD biology are excitatory (glutamatergic) and inhibitory (GABAergic) neurons, both of which can be generated in vitro from human pluripotent stem cells and depolarized to model the transcriptomic and epigenetic changes caused by neuronal activation. Our lab annotated the enhancers present in both cell types at baseline and activated states and found 2495 enhancers containing non-coding ASD DNVs, including several hundred that are cell-type specific or activity-dependent. Using a massively parallel reporter assay (MPRA), this proposal will determine whether non-coding DNVs found in individuals with autism alter cis-regulatory activity in glutamatergic or GABAergic human neurons in either baseline or activated states. Further, gene-enhancer mapping has revealed that a subset of DNV-containing enhancers is predicted to regulate genes previously implicated in ASD. To validate cis-regulatory activity and to compare trans-effects on downstream gene networks, a CRISPR inhibition screen will be performed in both cell types and activation states, targeting 25 ASD genes and their DNV-containing enhancers. If successful, this work will demonstrate the potential functional contribution of non-coding de novo variants to ASD biology, which has thus far remained an outstanding question in the field. Moreover, this will generate transcriptomic datasets for top ASD risk genes in two clinically relevant cell types at both resting and active states to expand upon the growing number of functional genomics ASD studies, emphasizing convergent regulatory gene networks. This research will take place at the Icahn School of Medicine at Mount Sinai, containing the 2nd best NIH-funded neuroscience department and home to the Seaver Autism Center, renowned for bridging basic science and clinical trials for more effective ASD care. The scientific rigor, innovative techniques, sophisticated analyses, multi-disciplinary collaborations, and ample mentorship opportunities outlined here would propel me towards a successful career as an independent research professor studying the molecular mechanisms underlying psychiatric disorders while mentoring future generations of scientists and advocating for mental health education.

NIH Research Projects · FY 2025 · 2023-04

Prenatal programming of child asthma and respiratory health is potentially influenced by maternal exposures, such as a woman’s lifetime stress, although mechanisms of this biologic embedding have not been fully delineated. Emerging evidence suggests that exposure to trauma can be a particularly robust potentiator of biological events that increase vulnerability to asthma in offspring and may help explain increased risk found in lower-income urban U.S. populations. Lower-income BIPOC (Black, Indigenous, People of Color) women experience traumas over their lifetime at rates above national U.S. samples. Research from our group has shown that lifetime exposure to traumatic stressors in women, even when remote, impact stress-related programming of respiratory disease starting prenatally. Oxidative stress (OS) resulting from an imbalance between reactive oxygen species (ROS) and antioxidant defenses is increasingly thought to play a central role in asthma pathogenesis and lung growth and development. While evidence indicates that BIPOC populations have increased OS, studies examining whether elevated OS, indexed using traditional biomarkers in prior studies, in part explains health disparities have been mixed. Inconsistent findings may be a consequence of select biomarkers used in prior studies. Moreover, the critical role placental OS plays in fetal programming is increasingly appreciated with a high reliance on mitochondrial function to maintain optimal oxidant balance. Chronic stress can result in dysfunctional mitochondrial processes and the accumulation of ROS-generating mitochondria. Thus, higher order biomarkers deployed in multiplex panels considered as complex mixtures and/or biomarkers of cumulative OS, may provide greater insight into underlying OS processes that vary across populations. Finally, emerging evidence suggests that relationships between OS and disease outcomes may be modified by underlying metabolic factors that vary by maternal race/ethnicity and body mass index (BMI). This proposal will leverage a well-established urban, ethnically mixed longitudinal pregnancy cohort study to examine associations among maternal lifetime stress, oxidative stress biomarkers, and children’s risk for repeated wheeze and asthma and reduced lung function by age 6-7 years assessing for joint effects of postnatal stressors and oxidative stress biomarkers. Maternal prenatal OS will be indexed by (i) a mid- pregnancy urinary oxidative stress panel (OS mixtures) and (ii) placental mitochondrial DNA (mtDNA) heteroplasmy. The proposed analyses will more comprehensively examine the role of OS in prenatal programming of child asthma and early childhood lung function including placental mitochondriomics. Accounting for modifying effects of maternal race/ethnicity and BMI may better inform observed disparities. In addition, elucidating molecular mechanisms may lead to novel prevention and treatment strategies and because of the central role of mitochondria in regulating the maternal-fetal interface, our findings may provide a model that can be extended to additional prenatal risk factors and other fetal disorders.

NIH Research Projects · FY 2026 · 2023-04

SUMMARY The gastrointestinal (GI) tract is the only abdominal organ that has evolved with its own enteric nervous system (ENS) fully contained within the gut wall, also known as the “second brain” in the gut. Our long-term goal is to understand how the intrinsic primary sensory neurons (IPANs) in the ENS detect and respond to both physical and chemical cues in the gut lumen and control propulsion of content in the colon. Although known for about 25 years, the IPANs are still a subset of the most mysterious neurons in the ENS because how they participate in coordinated muscle movements (motility), regulate immune cell function (immunity) and maintain integrity of intestinal barrier is not completely understood. Equally as mysterious is whether the IPANs can fulfil the function as a “pattern generator” and can control the rhythmicity of cyclical propagating contractions along the colon. This is largely due to a lack of tools that can be used to selectively manipulate the excitability of specific classes of enteric neurons and any drugs that have been tried to stimulate or block activity in IPANs will likely act on many other types of neurons (or non-neuronal cells), making interpretation of the results unclear. In pilot studies, we have generated critical resources enabling us to identify and selectively targeting the β- CGRP-expressing (β-CGRP+) IPANs. By using these unique resources, we will be able to ask important questions regarding the roles of the β-CGRP+ IPANs in the ENS: What role the β-CGRP+ IPANs have in the propagation of neural activity along the gut? Are these IPANs activated by both mechanical and chemical cues in the gut lumen? Can these IPANs serve as cellular sensors for distinct microbiota-derived metabolites? This proposal represents a major technical advance by using cutting-edge neurogenetic approaches which make it possible to genetically target and determine the functionality of the β-CGRP+ IPANs in the ENS both ex vivo and in vivo, providing the first insights into how selective activation and inhibition of the β-CGRP+ IPANs in the ENS affects GI-motility. This information will advance our understanding of the inner workings of the ENS and shed new insights on the development of novel strategies for the treatment of motility-related GI disorders by targeting the IPANs in the ENS.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Blunted or flat facial expression is characteristic of schizophrenia (Sz) spectrum disorders and their clinical high risk (CHR) states, and is associated with negative symptoms, social impairment, and poor outcome. Currently, there is no objective test to quantify blunted face expression in Sz. However, with advancements in computational methods, we can begin to operationalize blunted face expression, necessary for both methodological studies and clinical trials, especially as there are no evidence-based treatments. We propose computational analyses of time series of video frame-based estimates of movements of individual face muscles, based on Ekman and Friesen’s Facial Action Coding System or FACS, obtained during ecologically valid open-ended interview on a remote platform (and during a standard viewing paradigm of valenced stimuli). Face action units (AUs) index movement of individual face muscles, which have known physiology and circuitry. Our preliminary data are consistent with early small video coding and electromyography studies of face AUs in Sz, in replicating decreased mean amplitude of face AUs, especially of AU7 (“lid tightener”) or orbicularis oculi, which is involved in smiling, but also social signaling more broadly. Further, in generating matrix and distance profiles for face AU time series data, we find indices of decreased repertoire of face expression in Sz, also associated like amplitudes in CHR with ratings of blunted affect and poor function. Aim 1 is to assess the correlates and psychometric properties of these face expression amplitude and repertoire metrics in a large international cohort of early course Sz spectrum, CHR and healthy individuals well-characterized for demographics, symptoms, cognition and function, (and in whom effects of medications and chronicity may be less). These metrics are expected to differentiate psychosis from the norm, have convergent validity with ground truth ratings of “blunted affect”, and be correlated with social and role impairment, and also face processing ability. Variation by demographics (age, sex, ethnicity) will be assessed, as well as test-retest reliability at one year. Aim 2 is to align video and audio time series to create per frame sets of features and envelope metrics in segments in participants to test for differences in facial dynamics when individuals are speaking vs. listening. We hypothesize that global face expression will decrease significantly during pauses in individuals with Sz, consistent with preliminary data. Based on a cognitive model, we hypothesize decreased face expression and pause behavior will be correlated and associated with slowed processing speed. Aim 3 assesses synchrony of face expression between interviewers and participants, with patients hypothesized to have decreased synchrony and alignment across modalities. Overall, this large rich dataset of multimodal time series raw data will be archived and available for analyses, including more complex nonlinear time series analyses.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Mitochondria created an evolutionary advantage for eukaryote and metazoan organization, and their impact on cell biology extends from anabolic and catabolic metabolism to determining the final moments of cell survival by engaging apoptosis. Throughout the last decade, interest in studying how mitochondria influence cancer cell biology led our laboratories to identify mechanisms linking oncogenic signaling (i.e., BRAFV600E / NRASG12V ) to multiple mitochondria-centric processes within malignant cells including altered mitochondrial dynamics, oxidative phosphorylation, and chemosensitivity. More recently, we focused on exploring how oncogenes intersect upon mitochondrial biology prior to transformation – which will likely provide molecular details into pre- malignant cell biology and early stages of disease. We commonly position our studies in the context of melanoma as we have extensive experience with primary human melanocytes, integrated cohorts of patient RNA-seq datasets and tissues, and multiple in vitro and in vivo models of early and late disease. For instance, the introduction of oncogenic signaling (BRAFV600E / NRASG12/Q60) in primary human melanocytes causes rapid oncogene-induced senescence (OIS), and this tumor-suppressive mechanism is reflected in patients who present with pre-malignant skin lesions in the clinic. Therefore, the scientific premise for this application is based on three novel observations: (i) primary human melanocytes expressing oncogenes rapidly expand their mitochondrial networks during OIS; (ii) this expansion is dictated by the undescribed activation of the ATF5- dependent mitochondrial unfolded protein response (mtUPR); and (iii) the mtUPR controls the rate and extent of OIS. While the mtUPR is a fundamental organelle-specific quality control signaling pathway that is essential to mitigate mitochondrial stress, no literature mechanistically connects oncogenic signaling to mtUPR activation, melanocyte biology, nor melanoma progression. In our preliminary experiments, we explored biochemical signaling, mitochondrial responses, cellular gain-of-function / loss-of-function approaches, and hundreds of patient samples to establish the hypothesis that the oncogene-activated ATF5-dependent mtUPR is a key signaling pathway that instructs melanocytes during OIS and its escape. In this R01 application, we propose three complimentary, but distinct, specific aims to examine this hypothesis using melanocytes, numerous models of early disease, and patient samples. Specific Aim #1: Interrogate the mechanistic relationship between oncogenic signaling, mtUPR, and OIS. Specific Aim #2: Identify the gene expression programs mediated and maintained by the mtUPR during OIS and early disease primary melanoma. Specific Aim #3: Define the impact of mtUPR activation in models of nevi and primary disease.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Oncolytic viruses (OV) are an emerging class of therapeutics capitalizing on their preferential ability to replicate in cancer cells and modify the tumor microenvironment. Using Newcastle Disease Virus (NDV) as an OV model, we were one of the first groups to identify OV ability to induce tumor-infiltrating lymphocytes and synergize with immune checkpoint blockade. Based on this rationale, we have conducted a clinical trial of intraperitoneal (IP) ONCOS-102 (oncolytic adenovirus) in combination with anti-PD-L1 antibody durvalumab in patients with advanced ovarian and colorectal cancer (NCT02963831). Despite these studies, major gaps exist in our understanding of how OVs interact with the immune system. While an increase in TILs or “making tumors hot” is commonly used as a readout of response to OV, such readouts are highly nonspecific: (1) It is largely unknown whether the increase in TILs represents primarily tumor-reactive or OV-reactive T cells; (2) It is unknown whether the OV-reactive T cells positively or negatively impact anti-tumor immunity; (3) It is unknown how the balance between the tumor-reactive or OV-reactive T cells is influenced by biologically distinct OVs. In our preliminary studies, we find that intratumoral NDV markedly expands the number of T cell clones that are shared between the different tumor sites and that dominant T cell clones in the treated and distant tumors are associated with distinct T cell phenotypes, thus possibly identifying tumor- vs. OV-reactive T cells. Using immunologic and genetic tools such as T cell receptor (TCR) sequencing and single cell RNA-sequencing and our team’s expertise in computational biology, immunology and virology, the key objectives of our study are to define the balance between the OV-reactive and tumor-reactive T cells generated in response to OV therapy and to determine the impact of OV-reactive T cells on T cell-mediated anti-tumor immunity. In Aim 1, we will identify the distinct TCR sequences and phenotypes associated with OV-reactive and tumor-reactive T cells and will quantify their balance in the treated and distant tumors. We will determine how prior anti-OV immunity and OV-specific CD4 or CD8 T cells impact the anti-tumor T cell response and therapeutic efficacy. To ensure broad applicability of our findings, we will test our hypothesis in two distinct mouse models: flank B16 melanoma and peritoneal model of ovarian carcinoma MPB1 and will employ NDV (RNA virus) and vaccinia (DNA virus) as two biologically distinct virus models. In Aim 2, we will leverage the tumor microenvironment data as well as TCR data from pre- and on-treatment tumors and blood from our clinical trial of oncolytic adenovirus ONCOS-102 in combination with durvalumab to determine how putative OV- and tumor-reactive T cells evolve in tumors and in peripheral blood and whether these parameters are predictive of clinical benefit. This study will determine how different OVs activate anti-viral and anti-tumor adaptive immune response and will enable us to understand the nature of OV-induced T cell responses beyond “hot vs. cold” within the context of mouse and human studies. These findings will be key to understand the nature of immune responses to OVs in trials and to enable future development of strategies aiming to enhance anti-tumor rather than anti-OV immunity.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract: Opioid use disorder is a public health crisis in the United States with roughly 80,000 overdoses taking place in 2021. Addiction can be conceptualized in three phases: binge/intoxication, withdrawal/negative affect, and preoccupation/craving. Current treatments for opioid use disorder substitute reward and prevent withdrawal, but there exists a treatment gap for the habitual/compulsive use associated with the third phase of addiction, necessitating better understanding of the molecular and cellular underpinnings of this behavior. Using an unbiased screening of the epigenome of postmortem samples of dorsal striatum from human heroin users, the Hurd Lab identified a promoter region for the gene encoding Fyn kinase as the locus with the most variance explained by heroin use in neurons. Fyn mRNA expression was elevated both in these human samples and in the dorsal striatum of rats that underwent heroin self-administration. Inhibition of Fyn either with a small molecule drug or siRNA infusions into the dorsal striatum decreased response for heroin in a translational model of relapse. Different subregions of the dorsal striatum mediate different aspects of drug-seeking behavior. I found that rats that compulsively sought heroin showed increased Fyn expression in both dorsomedial and dorsolateral striatum, while rats that took heroin non-compulsively showed increased Fyn expression in the dorsomedial subregion. My project will investigate the molecular and cellular mechanisms of Fyn in heroin self-administration behavior. I will knock down Fyn in dorsomedial and dorsolateral striatal subregions and test motivated and compulsive heroin seeking behavior, with the hypothesis that Fyn knockdown in dorsomedial striatum will reduce goal-directed responding, while Fyn knockdown in dorsolateral striatum will reduce habitual responding. I will perform RNA-sequencing in these subregions to assess molecular networks modulated by Fyn. To determine how Fyn regulates neural activity during heroin seeking, I will record neuronal activity in the dorsal striatum using in-vivo fiber-photometry. These experiments will enable me to uncover the molecular and cellular mechanisms underlying Fyn’s role in heroin use disorder and its effects on behavioral models of motivated versus habitual drug taking. I will learn a behavioral model of compulsivity, calcium imaging, and RNA-sequencing which will equip me to answer questions relating molecular changes to neuronal activity to behavior in my future career as a physician-scientist.

NIH Research Projects · FY 2026 · 2023-04

(PLEASE KEEP IN WORD, DO NOT PDF) The long-term goal of this grant is to obtain a comprehensive understanding of the intercellular mechanisms involved in the maintenance of organismal heme homeostasis. In mammals, heme, an iron-containing cofactor and signaling molecule, is synthesized in all nucleated cells and is essential for a diverse array of fundamental biological processes. As a hydrophobic and cytotoxic molecule, heme must be transported in a highly controlled manner through membranes via specific cellular pathways. The current paradigm posits that mammalian cellular heme homeostasis is maintained solely by the balance between heme biosynthesis, driven by 5-aminolevulinic acid synthase (ALAS), and heme degradation. While the discovery of eukaryotic heme transporters has given rise to the emerging notion that inter- and intra-cellular heme trafficking pathways also contribute to the maintenance of heme homeostasis, the pathways and mechanisms that mediate heme homeostasis outside of the context of heme biosynthesis and heme degradation remain poorly understood. Our recent unpublished studies using a newly generated hepatocyte-specific ALAS knockout mouse model (Alas1/2KO mice) strongly suggest that hepatocytes acquire “on demand” heme from reticuloendothelial macrophages through a currently unexplored intercellular heme homeostatic pathway. In this proposal, we will test our paradigm-shifting hypothesis that hepatocytes maintain cellular heme homeostasis by acquiring a substantial portion of their heme from macrophages using the Alas1/2KO mice. To this end, Specific Aim 1 will identify the cell types that are directly involved in heme transfer from macrophages to hepatocytes by establishing an in vitro primary liver cell co-culture system. Specific Aim 2 will employ in vivo approaches to identify key molecules involved in the hepatocyte heme acquisition pathway and to investigate the mechanisms by which the acquisition system is activated. Specific Aim 3 will identify additional candidate genes and pathways that drive hepatocyte heme acquisition using single cell RNA sequencing. These studies will advance our understanding of how liver maintains heme homeostasis in health and disease and may aid in improving our understanding of the pathogenesis of the life-threatening acute neurovisceral attacks that occur in the acute hepatic porphyrias.

NIH Research Projects · FY 2025 · 2023-04

Project Summary Broadly neutralizing antibodies (bNAb) against HIV Env represent a novel class of antivirals that have the potential to neutralize viral entry, while simultaneously activating non-neutralizing immune effector functions. However, a major obstacle for their clinical use is identifying the virologically suppressed HIV+ patients that are best suited for a given bNAb or bNAb combination. Indeed, early-stage therapeutic studies with bNAb have not found uniform concordance between viral outgrowth assay sequences (in vitro) and the Env resistance mutations that arise in treated patients in vivo. Our laboratory has developed robust and quantitative flow cytometry-based assays for neutralization of both cell-free and cell-to-cell infection. We have observed decreases in the potency and efficacy of Abs using cell-associated versus cell-free HIV-1 neutralization assays. The detection of these cell-associated phenotypes may be particularly important for cure-based approaches where functional reactivity of bNAbs against infected cells may be important for in vivo efficacy. To overcome the limitations of PCR-first based approaches that often identify non-functional Env, we propose the development of a single cell Viral Outgrowth Neutralization Assay (scVONA) that will be performed in the presence and absence of bNAb. This scVONA will simultaneously quantify viral latent reservoir sequences and determine their sensitivity to bNAb. To accomplish this, scVONAs will leverage and enhance a highly sensitive single cell reporter cell line to detect infection and identify cells to test for neutralization of cell-to-cell spread. The ultimate readout are single cell flow cytometry measures that enable capture of resistant env genes by sorting and sequencing insensitive versus sensitive clones. Long read Env sequencing will be employed for bioinformatic analyses to determine linked sequences that correlate with resistance. Titration of bNAb will also provide estimated IC50 and sequences may allow a measure of the fraction of sensitive clones. We believe that scVONA will provide a rapid <2 week readout of neutralization sensitivity and simultaneously provide a measure of functionally intact env genes for confirmatory analyses, significantly reducing the time, cost and labor associated with identifying bNAb resistant sequences from HIV+ patient samples.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Myelodysplastic syndromes (MDS) are myeloid neoplasms with dismal prognosis, frequent progression to acute myeloid leukemia (AML) and no effective treatment. A decade ago, a development with transformative potential for this disease was the discovery that more than half of MDS patients have somatic mutations in genes encoding splicing factors (SFs, i.e. RNA binding proteins that regulate pre-mRNA splicing). While the high frequency and early occurrence during the disease course of SF mutations rendered them promising targets, efforts to therapeutically leverage this through splicing modulator drugs did not show promise in clinical testing and the drug development pipeline for these targets, and for MDS in general, is currently nearly empty. We developed genetically faithful human induced pluripotent stem cell (iPSC) models of SF-mutated MDS using CRISPR gene editing and performed integrative analyses of splicing (RNA-Seq) and RNA binding (eCLIP) to search for mis-spliced transcripts that are direct common targets of two of the main SF mutations (SRSF2 P95L and the U2AF1 S34F). We found that both mutant SRSF2 and mutant U2AF1 cause altered splicing of the gene GNAS, promoting the production of a longer isoform (GNAS-L), which in turn produces a longer form of the alpha subunit of the stimulatory G protein, G⍺s (G⍺s-L). G proteins are key signaling molecules involved in many important signaling pathways and cell functions, including oncogenic processes. Our preliminary data using functional, biochemical and population genetics approaches support a critical role for the long form of G⍺s (G⍺s-L) as an MDS driver and reveal a new mechanism by which SF mutations drive MDS that opens a completely novel and unexplored therapeutic avenue for MDS, AML and other cancers with SF gene mutations. The goal of this proposal is to investigate G⍺s as a therapeutic target for MDS and identify opportunities for therapeutic interventions that inhibit signaling by G⍺s-L. Specifically, we propose to: (1) Evaluate the effects of direct degradation (through a dTAG) or inhibition (by novel cyclic peptides) of G⍺s in splicing factor (SF)-mutated MDS and AML hematopoietic cells (primary and iPSC-derived) using recently developed iPSC-based models of MDS-to-sAML progression and longitudinal bone marrow samples from MDS patients who progressed to AML; (2) Characterize and target cell signaling downstream of Gs-L through candidate and unbiased approaches (Reverse Phase Protein Array, CyTOF, transcriptomics); and (3) Identify the G-protein coupled receptors (GPCRs) involved in Gs-L activation in SF-mutated MDS and AML through focused CRISPR KO screens. This study can establish a novel therapeutic target that may transform the treatment of MDS, AML and possibly other cancers.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Hepatic stellate cell plasticity and maladaptive fibrogenic memory in chronic liver disease Fibrosis associated with chronic liver disease affects hundreds of millions of patients worldwide. Hepatic stellate cells (HSCs), in turn, represent the main cellular driver of hepatic fibrosis. A key unanswered question is why HSCs activate to facilitate tissue repair in response to acute liver injury but hyperactivate to produce exuberant extracellular matrix in response to repeated liver injury, leading to fibrosis. Central to this behavior switch is a mechanism for HSCs to remember previous injury episodes in order to respond differently, i.e. hyperactivate, when exposed to re-injury. Recent studies, including ours (Wang et al. Dev Cell 2019), strongly support the epigenome and in particular DNA methylation patterns as the carrier of cell memory in development and in tissue injury. The objective of this research is to clarify how the epigenome and specifically DNA methylation patterns encode this maladaptive cell memory and amplify HSC’s fibrogenic response following re-injury. This proposal builds upon recent advances in single cell technology and low-input chromatin profiling which we optimized extensively to measure HSC gene expression and epigenomic changes in vivo in a novel mouse model of fibrogenic memory. In our fibrogenic memory model, HSCs completely deactivate following fibrosis regression, with their transcriptome indistinguishable from uninjured HSCs, however epigenomic changes persist in the form of chromatin accessibility changes. In response to re-injury, HSCs from regressed liver hyperactivate and are driven by unique transcriptional networks (WT1, TEAD1, TBX20, and PBX1) not found in HSCs undergoing initial injury. Using the Uhrf1 floxed mice generated previously in our Dev Cell paper to specifically remove Uhrf1, a critical component of the DNA methylation machinery, in HSCs, we found that these mice display augmented fibrogenic memory in response to re-injury. Our central hypothesis is that memory of previous injury through epigenetic changes modify HSC plasticity and amplify their activation in response to re-injury. We will address this hypothesis in three interrelated, but distinct specific aims:1) Define the regulatory elements controlling fibrogenic memory in HSCs; 2) Determine how UHRF1 and the DNA methylome control fibrogenic memory in HSCs; 3) Uncover novel regulatory nodes driving HSC’s maladaptive response in re-injury. This innovative approach leveraging cutting-edge genomics technology and unique animal models is significant because it will yield fundamental new insights into stellate cell biology by uncovering the epigenetic basis of fibrogenic memory, the contribution of specific genes and transcriptional networks to fibrogenic memory and hepatic fibrosis, and conserved fibrogenic drivers in liver disease patients that can lead to potential therapeutic targets.

NIH Research Projects · FY 2026 · 2023-04

Abstract A leading contributor to the significant mortality burden of prostate cancer, the second cause of cancer death among U.S. men, is the short-lived efficacy of androgen deprivation therapy (ADT), the mainstay of care for advanced and symptomatic disease. Increasingly alterations in DNA damage repair (DDR) genes, predominantly BRCA2, have been linked to ADT resistance and poor prognosis. We previously showed that deleterious alteration of BRCA2 is sufficient to induce ADT resistance in castration-sensitive prostate cancer (PC) cells. Our current proposal will investigate the molecular mechanism underlying BRCA2 loss/mutation- induced ADT resistance and progression to lethal prostate cancer. Using a panel of 107 DDR-associated genes from the PROREPAIR B cohort, we made the novel observation that ~82% of patients with mCRPC harbor alterations of one or more DDR genes. Herein, we aim to comprehensively investigate the role of DDR alterations (other than BRCA2) in the development of mCRPC and resistance to therapy. To do so, we will use CRISPR screening to prospectively investigate how the gain or loss-of-function alterations of DDR genes induce castration resistance. We have observed high expression levels of prostate-specific membrane antigen (PSMA) in response to the loss of BRCA2 and other DDR genes (e.g., ATM). Our second aim will investigate the impact of PSMA- targeted radiotherapy with [177Lu]-PSMA-617 and PARP inhibitor combination in BRCA2/DDR-deficient PC. Prostate cancer is predominantly resistant to immunotherapy. Since BRCA2-deficient cells also exhibit higher PSMA expression and are possibly immunogenic due to increased genomic instability, we will explore the effect of PSMA-targeted CAR T cells on BRCA2-deficient prostate cancer. Finally, as two PARP inhibitors (olaparib and rucaparib) have been approved to treat patients with DDR-deficient prostate cancer, we will investigate whether PARP inhibitors synergize with CAR T-cell therapy. For the first time, the proposed project will demonstrate the crucial importance of BRCA2/DDR alterations in prostate cancer biology and possibly lay the foundation for consideration of DDR alteration as the master driver of the transformation from indolent, localized prostate cancer to lethal mCRPC. We believe this work will lead to clinical trials that will establish new and effective treatments for this deadly disease.

Fonds de recherche du Québec – Santé · FY 2023-2024 · 2023-04

Volet: Renouvellement - Bourses de formation postdoctorale pour les citoyens canadiens et les résidents permanents; Domaine: Maladies infectieuses et immunitaires; Objet: Choc septique; Objet: Systèmes neuronaux; Application: Santé; Application: Évolution et traitement des maladies; Mots-clés: SEPSIS, INFLAMMATION, LOG-DISTANCE COMMUNICATION, BRAIN MOTOR CORTEX, NEUROIMMUNOLOGICAL CIRCUITS, HIGH-INTENSITY PHYSICAL ACTIVITY

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Alzheimer’s disease (AD) and related dementias (ADRD) are highly heritable, severe, and complex brain disorders. Molecular profiling efforts of the brain have enabled us to understand more about how common variants may be associated with disease risk via regulating gene expression. However, the status quo as it pertains to the use of multiomics data for gene-trait association, is limited to independent transcriptome- and proteome-wide association studies. Here, we propose to: (1) additionally leverage epigenetic datasets and (2) apply a data-driven correlation-aware meta-analytic approach to integrate a wide range of brain cell-type and area specific imputed transcriptomes, proteomes and epigenomes. Our preliminary data suggest that this will greatly increase the power to identify genetically driven gene dysregulation associated with these disorders while controlling for the correlation between different genomic features. First, we will impute brain cell-type (based on FANS/FACS-sorted and single-nucleus data) and brain area (PFC, ACC, EC, STG, IFG) specific transcriptomes, proteomes and epigenomes for AD, frontotemporal lobar degeneration, and dementia with Lewy bodies. Secondly, for each disease, we will perform a data-driven correlation-aware meta-analysis of these imputed genomic features to identify key genes. Lastly, we will develop an open resource that provides for the rapid dissemination and open access to analyses and outcomes. To our knowledge, the proposed approach will result in the largest multiscale modeling of genetically driven gene dysregulation in the brain, will identify novel genes associated with dementias and will enable similar studies in other brain disorders as well.

NIH Research Projects · FY 2025 · 2023-03

Project Summary Human genetic studies provide a powerful approach to identify genes and pathways associated with Alzheimer's disease (AD). Apolipoprotein E isoform 4 (APOE4) is the strongest genetic risk factor for sporadic Alzheimer's disease. Despite the strong effect of APOE4 on AD risk, a wide range of clinical outcomes exist within APOE4 carriers: some can develop cognitive impairment as early as 30 years of age while other carriers remain cognitively normal beyond 100 years. The large range in clinical manifestation amongst APOE4 carriers suggests that there are genetic variants that modulate APOE-associated risk. To investigate this hypothesis, we conducted stratified genome-wide association studies of APOE4 carriers at the extremes of age at onset distribution to identify variants that modify risk. We identified a single variant in EIF2B3 encoding the amino acid substitution S404A as a candidate risk modifier. eIF2B3 is a subunit of eIF2B, a guanine exchange factor (GEF) for eIF2 that is involved in translational control and the integrated stress response. Our lab has recently shown that APOE4 induces a constitutively upregulated ISR and aberrant protein translation in murine and human iPSC-derived microglia, demonstrating that APOE4 alone can lead to increased levels of cellular stress1,2. Furthermore, EIF2B2, another eIF2B subunit, is an AD risk gene identified in AD GWAS3. The overall goal of this proposal is to directly test the hypothesis that EIF2B3 variants modulate APOE4-associated risk by dysregulating the ISR and thus alter microglial cell function in the context of disease. In Aim 1, I will combine cell-free in vitro fluorescent GEF assays and structural chemistry to determine the effects of EIF2B3S404A directly on eIF2B structural integrity and enzymatic activity, in the absence of cellular stress. Aim 2 will determine the impact of EIF2B3S404A on APOE4 microglia cell function in vitro using human induced pluripotent stem cell (iPSCs)- derived microglia and live-cell imaging in the context of disease-related stressors. Lastly, in Aim 3, I will employ novel xenotransplantation methods involving direct injection of human microglia precursor cells into the mouse brain to evaluate the effect of EIF2B3S404A on APOE4 microglia cell function in vivo and in the context of disease. Together, the results here will confirm or refute the hypothesis that EIF2B3 variants modulate APOE4-associated risk by altering microglial cell function. Throughout the project, I will test the efficacy of an ISR inhibitor (ISRIB) drug at alleviating EIF2B3S404A -induced effects on EIF2B activity in vitro and in microglia in vitro and in vivo. The work here has the potential to greatly impact public health through understanding the way in which variants affect immune cell function and modify disease risk and by potentially identifying a novel drug for the treatment of AD. The fellowship training plan proposed here will provide me with the technical skills, scientific knowledge, critical thinking skills gained from Dr. Alison Goate's mentorship and the collaborative environment at the Icahn School of Medicine at Mount Sinai at large, equipping me with the necessary skills for a successful transition to a career as an independent scientist.

NIH Research Projects · FY 2026 · 2023-03

Myeloproliferative Neoplasms (MPNs) are chronic hematologic disorders that are associated with significant morbidity and mortality and have the potential to progress to myelofibrosis and a blast phase. These diseases are characterized by a series of driver mutations that originate at the hematopoietic stem cell (HSC) level, however, the mechanisms driving disease progression are currently unclear. Current therapeutics for MPNs largely do not alter the disease course and it is therefore imperative to decipher the mechanisms underlying progression in order to identify new, more effective therapeutic agents. This proposal focuses on elucidating the role of Phosphatase Mg2+/Mn2+ Dependent 1D (PPM1D) in MPNs and evaluating it as a potential target to prevent MPN disease progression. PPM1D is involved in the maintenance and differentiation of HSCs and has recently been found to be mutated and/or overexpressed in a subset of MPN patients. Preliminary data shows that PPM1D mutation/overexpression leads to increased fitness of the JAK2 mutated MPN clone and that PPM1D inhibition depletes MPN HSCs. To further discern the consequences of dysregulated PPM1D in MPNs we will complete the following aims utilizing a multipronged approach involving primary MPN samples, human induced pluripotent stem cell (iPSC) lines and murine models: (1) Delineate the effects of PPM1D dysregulation on molecular signaling in JAK2V617F+ human hematopoietic cells, (2) Assess the in vivo consequences of PPM1D overexpression on MPN disease phenotype, (3) Identify therapeutic vulnerabilities of MPN cells with dysregulated PPM1D function. Experiments will employ a variety of techniques including biochemical assays, murine transplant models, primary hematopoietic cell colony assays, RNA sequencing and genotyping of transcriptomes. This proposal will also evaluate a novel strategy to target PPM1D in MPNs. Dr. Bridget Marcellino, an assistant professor at the Icahn School of Medicine at Mount Sinai, will be completing these studies under the mentorship of Dr. Ronald Hoffman, a scientific leader in the field of MPNs. She will have 75% of protected research time and will be provided the necessary resources to complete these studies. As an institution Mount Sinai is ideal for fostering the development of physician-scientists with academic seminars and state of the art facilities and resources. Bridget will also have an advisory committee consisting of Dr. Eirini Papapetrou at Mount Sinai, Dr. Ross Levine at Memorial Sloan Kettering Cancer Center, Dr. Dan Avi Landau at Weill Cornell Medical College and biostatisticians, Dr. Amylou Dueck and Heidi Kosiorek at Mayo Clinic. The career development plan outlined here will allow Dr. Marcellino to gain scientific expertise as well as increase her skills in manuscript and grant preparation. This proposal will facilitate Dr. Marcellino to achieve both her short term goal of publishing her findings and her long term goals of developing as a physician-scientist and establishing an independent research program focused on identifying novel therapeutic targets in MPNs.

NIH Research Projects · FY 2026 · 2023-03

Dr. Karyn Goodman plays a critical role in Tisch Cancer Institute’s (TCI) cancer clinical trial program and in supporting NCI-funded clinical trials. She has demonstrated a long-term commitment to this work through her previous role as Associate Director (AD) of Clinical Research for 3 years at the University of Colorado Cancer Center. In the same role in the TCI, Dr. Goodman has been able to further expand on the work she was doing in Colorado and has focused on five major goals: 1) to expand access to clinical trials across the Mount Sinai Health System (MSHS); 2) to increase the numbers of NCI-sponsored trials performed across the MSHS; 3) to address the need for more oversight and integration of clinical trial services provided across the MSHS; 4) to support junior faculty to develop investigator-initiated trials (IITs) and present concepts to the Experimental Therapeutic Clinical Trials Network (ETCTN) for potential support; and, 5) to increase recruitment to TCI clinical trials. Institutionally, she has been the local PI of numerous National Clinical Trials Network (NCTN) studies at several NCI-designated cancer centers and is currently the Mount Sinai PI for one ECOG/ACRIN trial and one ETCTN trial and serves as the PI for the Early Drug Development Opportunity Program (EDDOP) at Mount Sinai. In addition, she is a national leader in the NCTN both through her involvement in the GI committees of the Alliance for Clinical Trials in Oncology and the NRG Oncology, as well as her role as the co-chair of the NCI Gastrointestinal Steering Committee. She has nicely bridged the work she has done nationally with her institutional AD of Clinical Research position. She brings her expertise in clinical trial development and oversight which she gained as the National Study Chair of the CALGB/Alliance 80803, a Phase II trial that evaluated PET-directed therapy for esophageal cancer, and the National Radiation Oncology Principal Investigator of the RTOG/NRG 0848, a large Phase III trial investigating the role of adjuvant chemoradiation in resectable pancreas cancer. Her overarching goal as a senior leader in the TCI is to support the development of both IITs and NCI-funded cooperative group clinical trials that will further advance the fight against cancer. The R50 funding mechanism will undoubtedly help foster her ability to expand NCI-funded oncology clinical research at Mount Sinai Hospital and expand access to these trials across the MSHS. Dr. Goodman is uniquely positioned for this award with both her institutional and national service focused on developing and supporting cancer clinical research.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT Bulimia nervosa (BN) is a disabling and often chronic eating disorder associated with medical complications and premature death. There is an acute need to identify the mechanisms that maintain BN and that may serve as targets for new treatments. Most studies have assumed that BN is perpetuated by stable, trait-like factors, including self-reported impulsivity and deficits in the ability to exert cognitive control. However, the out-of-control excessive intake and compensatory behaviors characteristic of BN are episodic and tend to alternate with periods of restricted intake or even fasting, suggesting intermittent oscillations in control. In addition, emerging work in behavioral economics and cognitive neuroscience suggests that failures to ultimately exert cognitive control depend on numerous neural computations, including: updating predictions about whether control will be needed in the next moment, and deciding whether trying to exert control is worth the costly cognitive effort. The overarching goal of this R01 is to clarify how these cognitive control computations may fluctuate across fasted and fed states in BN, potentially maintaining the cyclical nature of the disorder. Specifically, the proposed study combines functional magnetic resonance imaging (fMRI), computational modeling, and real-time mobile assessments to examine whether food consumption abnormally impairs frontostriatal function and associated control-related updating and effort-valuation processes in BN. We propose that the impairment of these processes in the fed state and relative improvement of these processes in the fasted state perpetuate out-of- control binge/purge episodes alternating with periods of dietary restriction. We will compare behavioral and neural responses of women with BN to those of group-matched healthy women during an inhibitory control paradigm to assess control-related prediction updating (Aim 1) and during a cognitive effort discounting paradigm to assess control-related effort valuation (Aim 2) in two states: after a 16-h fast and after a standardized meal. Aim 3 will use multi-modal symptom measures to examine main effects and potential interactions of state-specific updating and valuation processes on binge eating, purging, and restriction severity at baseline and at 6-month follow-up. The research team includes experts in BN-focused research, neuroimaging, computational neuroscience and psychiatry, advanced statistical methods, and ecological momentary assessment. The project innovatively 1) applies a neurocomputational framework to examine the roles of understudied subcomponents of cognitive control in BN; 2) assesses the influence of metabolic state (fasted, fed) on these subcomponents; and 3) relates the dynamics of these subcomponents in the laboratory to real-world, state-specific experiences and symptoms at baseline and over time. Data from this project will pinpoint altered elements of the control process that may represent prognostic biomarkers. Results will also identify the potential optimal patient state (i.e., fasted or fed) for control-focused interventions. Therefore, this study will yield vital data to inform urgently needed, precisely targeted treatments for cycles of binge eating, purging, and restriction.

NIH Research Projects · FY 2026 · 2023-03

Project Summary Axon regeneration after peripheral nerve (PN) injury is often incomplete. There is currently no effective treatment beyond surgical reconstruction, which is only beneficial for a small percentage of cases. Understanding the repair mechanisms is thus crucial. Here, we investigate a previously unknown function of the core circadian regulator Bmal1 in gating neuroregenerative responses after PN injury. The study is based on our new data that neuron-specific deletion of Bmal1 accelerates axon regeneration after PN injury. This exciting finding was made in the context of a series of advances from our laboratory in deciphering transcriptional networks that control neuronal intrinsic axon growth potential, i.e., how transcription factors (TFs) cooperate with epigenetic factors to reshape the chromatin landscape for induction of regeneration- associated genes (RAGs). Our most recent work has leveraged our genome-wide mapping of DNA hydroxymethylation (5hmC) dynamics in regenerating dorsal root ganglia (DRG) neurons. Intriguingly, we discovered enrichment of the Bmal1 binding motifs in genomic loci displaying 5hmC changes after PN injury, suggesting an interaction of Bmal1 with the 5mC/5hmC converting enzyme Tet3. Indeed, Bmal1 cKO in mice showed that the Bmal1-Tet3-5hmC axis regulates genes linked to axon growth, metabolism, and immune interactions. Moreover, pilot data show for the first-time a diurnal epigenetic rhythm of Tet3 and 5hmC in DRG neurons that is anti-phasic to Bmal1 transcriptional oscillation and corresponds to time-of-day differences in regenerative responses. Here, we will test the central hypothesis that Bmal1 functions as an inhibitor of axon regeneration and a gatekeeper of injury-trigged immune activation via regulation of 5hmC reprogramming. In Aim 1, we will characterize Bmal1-gated regenerative gene programs and examine the effect of pharmacological inhibition of Bmal1 transcription by SR9009, a potent Rev-Erb agonist with CNS penetration capability. Mechanistically, we will test a “two-hit model” wherein Bmal1 deletion primes DRG neurons, but an injury signal is required for RAGs induction. In Aim 2, we will characterize Tet3/5hmC epigenetic rhythmicity, correlation with “neurite outgrowth clock”, and the underlying mechanisms by testing the working model that Bmal1 controls Tet3 expression as well as Tet3 recruitment to target loci for 5hmC reprogramming. We will then map short- and long-term impact of PN injury on Bmal1-Tet3-5hmC rhythmicity and whether these rhythms return to normal upon axonal reconnection. In Aim 3, we pivot to in vivo study of the promoting effect of Bmal1 inhibition on nerve repair, including motosensory functions. We will also address sustainability of the effect, sex differences, and timed Bmal1 inhibition shortly after PN injury. In sum, our proposal has the potential of connecting Bmal1 circadian pathway, Tet3/5hmC epigenetic reprogramming, injury-triggered immune responses, and axon regeneration, thus advancing basic science of nerve regeneration and opening translational paths.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY SARS-CoV-2, the causative agent of the COVID-19 pandemic, modifies the cells that it infects in profound ways. One such modification is the activation of host cellular mitogen-activated protein kinase (MAPK) pathways, which contribute to severe inflammation that is a hallmark of severe COVID-19 disease. Inhibition of one MAPK pathway, the p38/MAPK pathway, reduces SARS-CoV-2 replication by an undefined mechanism. This proposal aims to measure the impact of human MAPK pathways on SARS-CoV-2 infection using a multidisciplinary approach that combines state-of-the-art proteomics technologies, medium-throughput genetic screening, and in vivo and ex vivo models of SARS-CoV-2 infection. These findings will inform the potential application of MAPK inhibitors for COVID-19 treatment and may identify alternative targets within the MAPK families. MAPK pathways play critical roles in many disease states, and this work will inform research in these areas by providing molecular mechanisms for MAPK regulation and providing tools for the unbiased discovery of MAPK substrates and regulators.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY/ ABSTRACT Psychosis typically emerges in late adolescence or early adulthood, which is a vital stage in social and cognitive development, and can therefore have a profoundly adverse impact on an individual’s long-term functioning. The onset of psychosis is preceded by a clinical high risk (CHR) phase characterized by attenuated psychotic symptoms and functional decline. CHR programs have enormous potential to reduce the long-term severity of the illness, and the suffering and cost associated with it. Youth at CHR also typically have environmental and individual-level barriers to accessing and engaging in services, including stigma, a dearth of trained providers, clinic location and transportation issues, suspiciousness, and a tendency to socially isolate. Reducing some of these barriers via telehealth interventions may improve treatment accessibility and engagement, thereby improving clinical outcomes. There is a substantial need to evaluate the feasibility of different CHR interventions to determine which may be most promising and for whom. There is also a significant need to systematically investigate remote delivery methods as a way of increasing access to critical services for CHR. We have established a comprehensive Group and Family-Based Cognitive Behavioral Therapy (GF-CBT) program for youth at CHR. GF-CBT aims to facilitate psychosocial recovery, decrease symptoms, and prevent or delay transition to psychosis in youth at CHR. GF-CBT is grounded in sociocultural ecological systems theory, psychosocial resilience models, and research on information processing in delusions. GF-CBT has been implemented as part of SAMHSA funded CHR services in New York, Missouri, and Delaware and has been adapted for telehealth delivery (GF-CBT-TH). This study will evaluate the feasibility and acceptability of GF-CBT-TH and gather data on the preliminary efficacy of GF-CBT-TH compared to individual CBT for CHR delivered via telehealth (I-CBT-TH). Subjects between the ages of 14 and 25 who meet CHR criteria on the SIPS (n=60) and their families will be randomly assigned to receive GF-CBT- TH or I-CBT-TH for a period of 15 weeks. Data will be collected at baseline, post-treatment, and 3-month follow-up. Feasibility of GF-CBT-TH will be measured by recruitment rate, session attendance, dropout rate, and subjects’ satisfaction with the interventions. The following intervention targets will be assessed in both groups: cognitive biases, social connectedness, family emotional climate, and family members’ proficiency in CBT and communication skills at post-treatment and follow-up. The GF-CBT-TH and I-CBT-TH groups will be compared across the following domains: psychosocial functioning, symptom severity, rates of remission from CHR, and rates of transition to psychosis. We will also explore whether patient treatment preference (for GF- CBT-TH vs. I-CBT-TH), family emotional climate and sociodemographic factors will differentially moderate treatment outcomes. In depth qualitative interviews will be conducted with patients, families, and clinicians to inform dissemination of GF-CBT-TH and make adaptations to the implementation manuals as needed.

NIH Research Projects · FY 2024 · 2023-03

Every year, 15 million babies are born premature. Over 75% of preterm births (PTBs) are termed spontaneous (sPTB) resulting in parturition at early gestational time points without clear causes. Our lack of understanding of the mechanisms and overall pathogenesis that promotes sPTB results in limited successful interventions. While uterine contractility and cervical remodeling appear to be obligatory processes in parturition, premature triggers of these processes remain poorly elucidated. Recent studies reveal close associations between cervicovaginal (CV) microbial communities and the occurrence of sPTB. In particular, we recently studied a cohort of 2000 pregnant women and assessed the CV microbial communities, metabolic and immune responses early in pregnancy, providing strong evidence that colonization with specific bacterial taxa, specific metabolic profiles, and local immune responses were strongly associated with sPTB. However, to develop preventive or therapeutic strategies, understanding the cause of sPTB is essential. We speculate that interplay between the CV microbial communities, local immune response and the cervical and vaginal epithelial barriers induce premature cervical remodeling and initiate sPTB. The overall goal of this study is to define how specific CV bacteria interact with vaginal and epithelial cells in clinically relevant in vitro and in vivo models and to understand how those interactions modify tissue remodeling and biomechanics of the pregnant cervix, driving sPTB. We propose a process whereby bacterial taxa that are highly associated with sPTB in humans provoke exfoliation of the vaginal epithelium. This process promotes epithelial-mesenchymal transition (EMT) from both vaginal and cervical epithelial cells. While activation of EMT prevents the ascension of these bacteria, a tradeoff is that EMT fosters breakdown of the extracellular matrix in the cervical tissue, triggering premature cervical remodeling and sPTB. Therefore, our central hypothesis is that specific bacteria, such as Gardnerella vaginalis (G.vaginalis) and Mobiluncus mulieris (M. Mulieris), promote EMT of the vaginal and cervical epithelial barrier which alters the structure and function of the pregnant cervix, leading to sPTB, even in the absence of ascending infection (above the cervix). This paradigm-shifting hypothesis will be tested through a series of in vitro and in vivo experiments. This proposal will first address whether ascension of bacteria into the uterus is actually necessary for PTB to occur; these studies have the potential to reframe our scientific and therapeutic approach to PTB. We will then demonstrate how bacteria induce EMT in CV epithelial barriers and how EMT might promote premature cervical remodeling. Unique to this proposal, we will provide quantitative assessment of the pregnant cervix, in terms of structure and function, in a mouse model of PTB. A multidisciplinary team adds rigor to our work by applying novel concepts and techniques to the study of sPTB. These studies will provide insight as to new and focused therapeutic targets to limit or prevent sPTB and will significantly advance this field.

NIH Research Projects · FY 2026 · 2023-03

The rate of aging of the breast differs between women. Further, older women tend to develop estrogen receptor alpha (ERα) positive breast cancer sub-type, despite lower hormonal levels. Our preliminary data compel us to hypothesize that mitochondrial genetics alters the rate of aging of the breast and impacts the sub- type of breast cancer. The mammary ductal tree is composed of luminal hormone sensitive (HS), ERα positive cells as well as luminal alveolar (AV) and basal cells that are both ERα negative. A recent scRNAseq analysis revealed that in both the basal and luminal AV cells, mitochondrial function declines with age but this decline was not observed in luminal HS cells. Rather, the luminal HS cells seem to up-regulate of the unfolded protein response of the endoplasmic reticulum (UPRER). The UPRER is closely interconnected with the mitochondrial UPR (UPRmt). Our group has identified the ERα and the mitochondrial sirtuin-3 (SIRT3) as key players of the UPRmt. While the level of the ERα does not fluctuate with age, in most individuals, SIRT3 levels decrease with age. Therefore, our central hypothesis is that the decline in mitochondrial function observed in the basal and luminal AV cells over aging may be due to the decline in the SIRT3 axis of the UPRmt. However, in the luminal HS cells this decline is not observed as they maintain mitochondrial function through the ERα axis of the UPRmt. We hypothesize that the ability of luminal HS cells to maintain mitochondrial function through aging, allow them to survive transformation and explains the selective bias toward ERα positive breast cancer in older women. Further, we performed RNAseq on the young and old-females derived mammary tumors and established luminal HS cells derived from both young and aged females. We found that markers of the ER? axis of the UPRmt and UPRER are up-regulated specifically in the aged luminal HS cells. This observation suggests that the transcriptional program of the ER? may be altered by aging. We hypothesize that the increase in ROS and the decline in hormones during aging alter the transcriptional program of the ERα. Further, the rate of decline in SIRT3 with age varies between individuals. Likewise, we found that the levels of SIRT3 differ between the BL/6NZB and BL/6C57 mice which have the same nuclear genome (BL/6), but different mtDNA; (C57 or NZB). Therefore, the implication is that the rate of decline of the SIRT3 axis of the UPRmt with age differs based on mtDNA haplotypes. Lastly, we hypothesize that while BL/6C57 mice (low SIRT3) will develop exclusively ERα positive mammary tumor over aging, in BL/6NZB females (high SIRT3) both basal and ER? positive mammary tumors will be observed. To test these hypotheses, we propose the following aims: Specific aim 1: Analyze of the UPRmt and UPRER and the ERα transcriptome in ERα positive luminal mammary cells over aging. Specific aim 2: Perform scRNAseq analysis of the mammary gland over aging in BL/6C57 and BL/6NZB mice. Specific aim 3: Compare the sub-types of mammary tumors between BL/6C57 and BL/6NZB mice over aging. We propose to do these analyses in pre-, peri- and post-menopausal as well as elderly mice.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY The goal of Dr. Neha Goel’s K23 project is to enable her to become an independent physician-investigator who conducts research on the implementation of evidence-based care for patients with acute respiratory failure. Building upon her strengths as a pulmonary-critical care physician and her prior training in clinical emergency care research, Dr. Goel will transition into the next phase of her career through a well-defined training plan. The training plan is meticulously designed to enable her to acquire requisite skills in guideline development, implementation science methodology, and advanced statistical analyses. Her career development plan contains focused training through conduct of a mentored research study, relevant didactic coursework, and close mentorship from leaders in the fields of emergency and critical care medicine, and implementation science. Dr. Goel’s ultimate research objective is to improve clinical outcomes for patients with acute respiratory failure through the implementation of a systematically developed, evidence-based, high flow nasal oxygen (HFNO) monitoring protocol for the emergency department (ED). In keeping with this objective, her K23 mentored research project will focus on investigating the implementation of evidence-based protocol on HFNO use and management for The Mount Sinai Hospital ED. HFNO use and management is a timely and important research topic. Since the COVID-19 pandemic began, there has been a rapid increase in HFNO usage. However, guidance on HFNO use and monitoring in the ED setting is lacking, and adverse clinical outcomes occur in the absence of guidance. Dr. Goel’s K23 research project specific aims are to: (Aim 1) develop a pragmatic protocol for HFNO use and management in ED setting for hypoxemic acute respiratory failure using feedback from a national panel of multi-disciplinary experts and current evidence; (Aim 2) select implementation strategies that address the determinants of protocol-concordant HFNO use and management, commensurate with the ED’s capabilities, resources, and needs of its patients; and (Aim 3) examine both the implementation process for the HFNO protocol and the applicable clinical outcomes in a pilot hybrid effectiveness-implementation study. The completed project will include an implementation plan for HFNO management in the ED setting, and will provide estimates of effect and sample sizes for a future R01 implementing and testing the impact of HFNO monitoring protocols across health systems. Using the skills and preliminary data gained throughout this K23 mentored career development project, Dr. Goel will be well positioned to become an independent physician-scientist with R01 funding to implement and test the effectiveness of respiratory care protocols for improving patient and process outcomes across health systems. Larger scale implementation of such interventions can help standardized care delivery, with the ultimate goal of improving patient outcomes for more than a million acutely ill patients presenting to EDs annually in the United States with respiratory complaints.