Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2025 · 2025-09

Aberrant actin-thin filament assembly results in life-threatening muscle diseases such as dilated cardiomyopathy (DCM) and nemaline myopathy. Our long-term goal is to determine the physiological link between thin filament architecture and myopathies, with the ultimate goal of formulating novel therapeutic strategies. It is known that sequential exchange of three α-actin isoforms: α-smooth (α-SMA), α-skeletal (α-SKA) and α-cardiac (α-CAA) into thin filaments is critical for development and function of striated muscle, and perturbations in this process lead to altered contraction and myopathies. However, the mechanisms underlying regulation of this process remain largely unknown. We discovered actin-binding cyclase associated protein 2 (CAP2) is a major regulator of cardiac thin filament assembly. Specifically, we found that it mediates α-actin isoform exchange by both regulating cofilin-2’s ability to depolymerize filaments comprised of α-SMA and promoting preferential incorporation of α-CAA into thin filaments; in other words, CAP2 acts as novel “actin monomer exchanger” of thin filaments. Accordingly, we found that cardiac thin filaments from Cap2-knockout (KO) mice contain low levels of α-CAA and high levels of α-SKA and α-SMA, while normally α-SKA and α-SMA are down-regulated in hearts of wild type mice as they age. These findings are important since CAP2 mRNA levels are reduced in human cardiomyopathies and the protein is critical for life; human mutations in CAP2, as well as knockout of Cap2 in mice, lead to DCM and sudden cardiac death. Unexpectedly, we discovered that CAP2-mediated subcellular alterations, and cardiac dysfunction are significantly more prominent in male mice, which may help to explain why human idiopathic DCM is more often frequent in men, with earlier age of onset and higher mortality. Together, these data lead us to an overarching hypothesis that aberrant α-actin isoform exchange exacerbates cardiac disease progression, and restoring proper actin isoform expression in myopathic hearts will ameliorate disease development. Excitingly, in support of this hypothesis, we found that expression of GFP-α-CAA in Cap2-KO hearts via adeno-associated virus (AAV) remarkably ameliorates onset of cardiomyopathy. We propose a multidisciplinary approach utilizing a unique combination of state-of-the art in vitro biochemical, biophysical and structural assays, cellular experiments, and novel CAP2 murine models to accomplish Specific Aims to determine: the role of CAP2/cofilin-2 complex in thin filament assembly and contraction; the role of individual CAP2 domains in -actin isoform switch; the relationship between thin filament organization and sex-specific cardiomyopathy; and whether modulating α-actin expression in diseased hearts improves cardiac differentiation and contractility. This work is of broad interest since abnormal α-actin isoform switching is a hallmark of many myopathies, regardless of the underlying cause of disease. This proposal leverages our ~30 years of experience filling critical gaps in our understanding of muscle contraction in health and disease.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY: CCR5 is a chemokine receptor involved in leukocyte trafficking. We identified a neuroprotective role for CCR5 in the context of West Nile virus (WNV) infection both in mice and humans. In mice, we found that mice lacking Ccr5 are significantly more susceptible to WNV-induced encephalitis compared to wild type (WT) mice. We translated these results using human cohorts, where we found that individuals that naturally lack CCR5 (CCR5∆32 homozygotes) develop more severe symptoms following infection with WNV. Furthermore, we and others have shown that this protective effect was not limited to WNV infection, but that the loss of CCR5 increased susceptibility to numerous other neurotropic flaviviruses, including tick-borne encephalitis virus (TBEV), and Japanese encephalitis virus (JEV). ZIKV is of particular concern because of its impact on pregnancy, with microcephaly and other congenital malformations, as well as fetal loss, stillbirth, and preterm birth among the possible outcomes. In preliminary data, we found that the loss of CCR5 increases susceptibility to ZIKV (a known teratogen) during maternal infections, resulting in poor pregnancy outcomes. This was also true for WNV, a virus not known to impact pregnancy. In this application, we will test the hypothesize that CCR5 is protective against flavivirus-induced congenital disease. This application will unravel the mechanisms involved in CCR5-mediated protection during maternal flavivirus infections.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Coronary heart disease (CHD) is a leading cause of death in the U.S. Despite the importance of diet as a primary prevention strategy, current population-level dietary guidelines are ineffective, resulting in rising disease prevalence. Precision nutrition can improve CHD prevention but is limited by our inability to accurately predict what dietary pattern(s) will associate with disease at an individual level. To realize the promise of precision nutrition, two problems with the current approaches to diet modeling must be addressed: the modeling of complex dietary patterns and measurement of dietary intake. Traditional dietary pattern analysis methods oversimplify dietary components and cannot capture complex food interactions, and self-reported dietary intake is prone to recall bias. The application of artificial intelligence to large-scale biobanks offers a potential solution. Variational autoencoders (VAEs) and graph neural networks (GNNs) are classes of deep learning models that learn a simplified representation of high-dimensional data and have the capacity to model complex relationships, such as those present in dietary intake data. Under the guidance of a mentorship team with expertise in precision medicine and clinical informatics, I propose to apply these innovative methods to the UK Biobank (UKB; ~103,000 individuals) and the National Heart, Lung, and Blood Institute (NHLBI) Trans-Omics for Precision Medicine (TOPMed; ~30,000 individuals) cohorts. I will use a GNN to model compositional dietary patterns and a VAE to model omics–dietary pattern associations as objective markers of dietary intake. To address limitations in dietary pattern modeling, I propose: Aim 1 – Evaluate the utility of GNNs to model dietary patterns and food interactions. I will perform a comparative analysis of GNNs against principal components analysis and k-means clustering, current data-driven dietary pattern analysis methods, by evaluating (1A) the consistency and novelty of dietary patterns identified across methods, (2B) the accuracy of CHD risk prediction; and (1C) robustness of the model through external validation in TOPMed. I hypothesize that the GNN will (1) identify novel dietary patterns, (2) model the impact of food interactions on CHD risk, and (3) outperform traditional methods in CHD risk prediction with robust results across datasets. To address limitations in dietary intake assessment, I propose: Aim 2 – Identify multi-omics profiles of dietary patterns as an objective marker of dietary intake. I will apply the previously developed multi-omics variational autoencoder (MOVE) framework to UKB and TOPMed cohorts to (2A) identify novel metabolomics/proteomics profiles that associate with dietary intake patterns, and (2B) characterize individual response patterns to diet perturbation. I hypothesize that (1) MOVE will identify novel multi-omics response patterns of dietary intake and (2) there will be distinct omics and clinical response patterns to diet perturbation. This project, supported by the Icahn School of Medicine, offers training in precision nutrition and nutritional epidemiology, facilitating my professional development and computational skills expansion for an independent research career.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Obsessive-compulsive disorder (OCD) is characterized by repetitive thoughts and behaviors, often driven by an inability to adapt behavior in response to changing environments. The basal ganglia, a network of subcortical nuclei, plays a crucial role in cognitive and motor processes, and imbalances between the direct and indirect pathways of this network is implicated in OCD. The indirect pathway is hypothesized to be hyperactive in OCD, leading to excessive suppression of behavioral flexibility. This hyperactivity may underlie the hallmark compulsive and repetitive behaviors seen in OCD, which can be interpreted as an over-reliance on habitual associations, effectively "locking" patients into rigid behavioral states This project aims to investigate the role of the external part of the globus pallidus (GPe), a key node in the indirect pathway of the basal ganglia, in compulsive choice behavior. By leveraging intracranial single-unit recordings in OCD patients undergoing deep brain stimulation (DBS) surgery, this study will assess how neural activity in the GPe contributes to adaptive and maladaptive decision-making during a reversal learning task. The goals of this study are twofold: (1) characterize the role of the GPe in adaptive and maladaptive decision-making in OCD, and (2) determine the computational contributions of GPe to the basal ganglia indirect pathway using single- unit recordings during reversal learning. Specifically, we hypothesize that neural activity in the GPe will be modulated during changes in choice selection ("switch" versus "stay" conditions), with increased firing rates observed during switch conditions. Additionally, we predict that GPe activity will differ based on the outcome valence of decisions (positive, negative, and neutral results). We will also test the hypothesis that reward prediction errors and choice confidence, estimated using reinforcement learning and Bayesian models, respectively, will be correlated with GPe firing rates. By addressing these aims, this project will generate novel insights into human GPe signaling, which has never before been recorded, offering a unique perspective on the neurobiology of compulsive behaviors. Ultimately, these findings may lead to new therapeutic targets for OCD, improving our understanding and treatment of this debilitating disorder. This research will also provide the applicant with comprehensive training in computational modeling and human electrophysiology, preparing her to become a leader in computational psychiatry.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Over 40 million people in the U.S. live with serious illness, many of whom experience significant financial impact due to the high costs, utilization, and risk of mortality that accompany serious illness. Older adults in particular have a greater number of chronic conditions and lower income to manage increased health spending as they retire. One quarter of older adults experience out-of-pocket spending (OOPS) in the last five years of life that exceeds their entire household assets. Despite the disproportionate impact of high OOPS on older adults with serious illness, the underlying drivers of high OOPS, and the potential care consequences from high OOPS for this population, are inadequately understood. Prior studies show drivers of higher OOPS for seriously ill older adults include serious illness type, health care utilization, insurance type, and living situation, among other factors. However, none have examined how insurance generosity affects high OOPS for seriously ill older adults. Insurance generosity varies within insurance types and can be dictated by state-level policy. Omitting insurance generosity from models of high OOPS excludes a highly modifiable factor that could mitigate high OOPS and their consequences. Such consequences of high OOPS include potentially burdensome care (e.g., emergency department reliance, days away from home, care transitions in the last three days of life, and in-hospital death), some of which have been associated with lower care quality at end-of-life as rated by patients and families; and depression, which has a well-established relationship with decreased wealth. Examining insurance generosity at the onset of serious illness, when OOPS is typically most concentrated, may further illuminate a path through which potentially burdensome care, depression, and care quality could be addressed. In this project, we will leverage rich health and financial data from the nationally representative Health and Retirement Study (HRS) to a) determine the relative contributions of insurance generosity, serious illness type, and other factors to high OOPS among Medicare beneficiaries over 65 with new-onset serious illness, and b) test the association of insurance generosity during serious illness onset with subsequent burdensome care and depression among Medicare beneficiaries over 65. Such analyses will provide a more complete picture of the factors contributing to high OOPS for a population that is most affected, and also reveal novel insights on the relationship of insurance generosity with burdensome care and depression. These data can guide researchers, health systems, policy makers, and the public towards consideration of insurance generosity as a possible target for intervention. Mitigating high OOPS and its consequences at serious illness onset has the potential to interrupt a cascade of wealth depletion, worsened depression, health decline, and escalating care needs for older adults with serious illness.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY The overall goal of this proposal is to delineate epigenetic mechanisms that control Meibomian gland homeostasis and are dysregulated in aging. The Meibomian gland’s key function is to secrete a lipid-rich film that coats the ocular surface and prevents tear film evaporation. Aged Meibomian glands show reduced size and decreased proliferation of basal progenitor cells. These phenotypes are thought to contribute to Meibomian gland dysfunction in aging, including development of Evaporative Dry Eye Disease (EDED) which is common in aged patients, can seriously impair vision and currently has no effective treatment. Histone deacetylases (HDACs) are druggable targets with therapeutic potential and play key roles in controlling progenitor cell activity and maintenance in other epithelia. HDACs function by modifying chromatin and by controlling the activities of key transcription factors. Our published data show that inducible Hdac1 and Hdac2 deletion in adult Meibomian gland basal cells causes decreased proliferation and Meibomian gland loss. The transcription factor GLI2, a key component of the Hedgehog pathway, is hyper-acetylated in Hdac1/2 mutant Meibomian glands, a modification that is known to reduce GLI2’s transcriptional activity and association with active chromatin. Deletion of the Hedgehog co-receptor Smo causes decreased basal proliferation and Meibomian gland hypoplasia, partially mimicking Hdac1/2 loss, while epithelial deletion of the Hedgehog inhibitory co-receptor Ptch1 or forced expression of activated GLI2 in Meibomian gland basal cells results in acinar basal cell expansion, identifying Hedgehog signaling and GLI2 as key regulators of Meibomian gland proliferation. Comparative analysis of single nuclear RNA-seq data from the eyelid tarsal plates of young and old mice showed that Hedgehog signaling is decreased in aged compared with young Meibomian glands. This decline is associated with increased acetylation of GLI2 and histone H3, suggesting that HDAC1/2 activity is decreased in aged Meibomian glands. By contrast, Hdac3 deletion increases basal progenitor cell proliferation and GLI2 expression but not its acetylation levels, like the effects of elevated Hedgehog signaling. Based on these data we hypothesize that HDAC1/2 and HDAC3 play distinct and key roles in adult Meibomian gland progenitor cells in part by differentially modifying the expression and activity of Hedgehog pathway components and target genes, and that decreased HDAC1/2 activity contributes to aging phenotypes. To test this hypothesis and delineate additional mechanisms in an unbiased fashion, we will use genetic and single- cell genomics approaches to identify the direct targets and transcriptional co-regulators of HDAC1/2 and HDAC3 in Meibomian gland progenitor cells and determine the extent of overlap in HDAC1/2-regulated genes with those dysregulated in aging. These experiments have potential to reveal novel therapeutic strategies to enhance progenitor cell activity in aged Meibomian glands.

NIH Research Projects · FY 2025 · 2025-09

Liver transplantation stands as the sole curative remedy for individuals grappling with end-stage liver disease. The scarcity of donor livers poses a formidable obstacle, compounded by the substantial number of discarded organs due to viability concerns. The advent of Normothermic Machine Perfusion (NMP) during the past few years has opened a new era by employing machine perfusion to enhance liver graft viability. Normothermic perfusion has specifically emerged as a revolutionary technique, involving the assessment and reconditioning of donor livers at normal body temperature (35-37°C). This process extends liver viability and augments transplant success. While normothermic perfusion holds immense promise, it is currently hindered by the lack of comprehensive data and standardized protocols. We propose to establish an international cohort of NMP cases using a tailored digital data collection, storage, and sharing system under the PUMP consortium, the PUMP platform, and leverage machine learning and artificial intelligence to standardize protocols, identify key markers of liver viability, and predict post- operative outcomes. The project has two main objectives: to establish a international cohort of NMP cases through the usage of the PUMP platform (Aim 1) and developing a machine-learning engine to predict post-operative outcomes using the registry data (Aim 2). To achieve this, we have put together an interdisciplinary team of liver transplant surgeons, statisticians and machine- learning experts who have worked together during the past year to build the environment that will sustain the proposed project. Leveraging the robust statistical power derived from the aggregation of data from thousands of perfused liver transplants worldwide, our ultimate objective is to increase liver availability, mitigate complications, enhance patient outcomes, and provide personalized clinical decision support for physicians caring for liver transplant patients. This proposal holds the promise to fundamentally reshape the landscape of liver transplantation, ushering in advancements that stand to significantly enhance the lives of thousands of individuals awaiting liver transplants.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Advances in immune checkpoint inhibition (ICI) therapies have shown great promise in activating lung adenocarcinoma (LUAD) patients’ own anti-tumor immunity and prolonging survival. However, such immunotherapies have shown variability in response among LUAD patients of different ages. With the increasing age of lung cancer patients at diagnosis (35% are >75), there is a critical need for better understanding of the relationship between age and the LUAD tumor microenvironment (TME). New insight into the age-related cellular and molecular mechanisms and pathways in LUAD have the potential to uncover new therapeutic opportunities to modulate the TME, synergize with existing therapies, and improve disease outcome in elderly patients. We propose to use highly-parallelized single-cell RNA-sequencing (scRNA-seq) and emerging single-cell resolution spatial technologies to characterize the effect of age on the LUAD TME. We will integrate the scRNA-seq data collected as part of the Human Tumor Atlas Network (HTAN) with publicly available data to increase our cohort size to ≥178 LUAD patients. The expanded scRNA-seq data set will provide us with the necessary statistical power to decipher the cell type expression, cell composition, and regulatory differences between LUAD patients of different ages. We will also use machine learning to model the effect of cell type specific gene and pathway expression on TME cell compostion and search for therapeutic targets across the integrated cohort in order to better understand the underlying immunomodulatory mechanisms in the LUAD TME and how they associate with age. Finally, we will validate our computational modeling by measuring the spatial organization of the TME using tissue samples from LUAD patients of different ages. Our study promises to improve our understanding of the effect of age on cell-cell interactions, gene expression programs, and multicellular communities in the TME, identify new treatment strategies, and uncover new biomarkers that can improve therapy decisions for elderly LUAD patients. Finally, our general methodology can be extended to study the relationship between age and the TME in other tumor types in order to discover new age-specific therapeutic targets and improve patient stratification for treatment in different disease contexts.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Advancing personalized medicine requires the study of women's health encompassing female-specific condi- tions, such as adverse reproductive outcomes, endometriosis, menopause and gynecological cancers, as well as disorders that clearly differ in prevalence, etiology and natural history between sexes, such as chronic respir- atory disease, cardiometabolic disease, and neuropsychiatric disorders. The underlying pathogenesis of multi- factorial disorders with variable onset across the lifespan reflect development-specific exposures/experiences, at both the individual- and broader community-level, and individual response to these factors. Programming of health outcomes results from environment-induced shifts in a host of integrated molecular, cellular, and physio- logical states against a genetic background with innumerable social and chemical, nutritional, and microbial exposures modulating these mechanisms. Moreover, sex-specific biology further modulate disease trajectories over the lifespan. Given the cross-disciplinary contributing factors, addressing women's health will require transdisciplinary (TD) team-based science training that integrates and extends beyond discipline-specific concepts, approaches, and methods. Life course science underscores the relative importance of exposures dur- ing different life stages in relation to health, with exposure(s) within vulnerable windows having more significant and lasting effects on health than those outside these windows. Although much focus has been on the in utero and early childhood periods in disease programming, other life stages including adolescence (puberty), pregnancy, and the menopause transition, need to be considered as vulnerable windows during which future women's health potential is programmed or shifted. These complexities underscore the need to conduct studies addressing both susceptibility windows and the importance of coincident exposures to elucidate disease mani- festations. The exposome concept addresses such complexities as it frames the study of the effects of all health- relevant environmental factors over the life course using both targeted and untargeted (omics-scale) discovery approaches. In light of the institutional investments, established infrastructure and resources, as well as men- tored training track record summarized herein, Mount Sinai Life-course, Exposomics, and Analytic Program (LEAP) in Women's Health K12 is uniquely poised to constitute a unique training ecosystem for women's health scholars focused on developing transdisciplinary (TD) competencies and foundational principles in life course theory, exposure science and environmental epidemiology, and data science - all key components of exposomics - to address current research and training gaps in women's health. Proposed initiatives will be facilitated through the LEAP Core with centralized resources and scientific expertise that establishes a knowledge base serving to accelerate training in cutting-edge research that more comprehensibly considers environmental influences on women's health across the life-course.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY This application outlines a program of research designed to better understand the molecular, cellular, and circuit- based mechanisms of opioid use disorder (OUD). We recently showed that μ opioid receptors (μORs) are densely expressed by neurons in the dorsal peduncular nucleus (DPn) of the ventral prefrontal cortex. Genetic ablation of μORs from DPn neurons abolished the rewarding effects of opioids and instead rendered opioids highly aversive. Disrupting μOR signaling in the DPn also exacerbated the severity of physical and affective components of the opioid withdrawal syndrome, while chemogenetically silencing neural activity in the DPn attenuated opioid withdrawal. Single-cell and spatial transcriptomics showed that μORs in the DPn are expressed by an atypical population of cortical pyramidal neurons that also express vesicular glutamate transporter 2, which densely innervate the parabrachial nucleus (DPnvGlut2àPBn neurons). These exciting new findings suggest that DPnvGlut2àPBn neurons play critical roles in the behavioral actions of opioids that drive the development of OUD. In this application, we will use cutting-edge molecular, cellular, and behavioral approaches to investigate the role of DPnvGlut2àPBn neurons in the motivational properties of opioids. In AIM 1, we will use intersectional genetics strategies to manipulate the expression of μORs in only DPnvGlut2àPBn neurons and thoroughly investigate the consequences on intravenous (IV) opioid self-administration behavior. In AIM 2, we will use whole-cell electrophysiological recordings combined with single-cell sequencing (PATCH-seq) to define the molecular identity of the PBn neurons that receive direct excitatory synaptic input from DPnvGlut2 neurons (PBnDPn neurons). We will then chemogenetically manipulate the activity of these PBnDPn neurons and determine the consequences on IV opioid self-administration behavior. In AIM 3, we will use single-nuclei RNA sequencing (snRNA-seq), single-nuclei spatial transcriptomics, and whole-cell electrophysiological recordings to define the transcriptional and function adaptations that occur in DPnvGlut2 and PBnDPn neurons during the development of opioid dependence. We will also investigate the behavioral consequences of chemogenetically manipulating the activity of DPnvGlut2 and PBnDPn neurons on the expression of physical and affective components of the opioid withdrawal syndrome. This highly innovative program of research builds on exciting new findings from our laboratory and promises to yield fundamentally new insights into the neurobiological mechanisms of opioid addiction.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Stress-induced disruptions in brain and heart communication can perturb the balance between sympathetic and parasympathetic responses, potentially causing health consequences across multiple domains. Our laboratory has focused on understanding the mechanisms of organ-organ communications under stress via two pathways: vagus nerve-to-brain and brain-to-heart. The vagus-to-brain communication involves the nodose ganglion, transmitting sensory interoceptive information such as heart rate to the nucleus of the solitary tract (NTS), the brain's visceral center. The parasympathetic regulation of the cardiovascular system occurs through the nucleus ambiguus (NAmb) brain region, a crucial brain-to-heart communication circuit. The Principal Investigator (PI) has made significant strides in investigating brain and heart interactions through pilot studies. Our preliminary findings show that ethologically relevant stressors negatively affect these two parasympathetic regulations, leading to disruptions in behavior, cardiorespiratory, immune, and other functions. We have further discovered the role of the neuropeptide pituitary adenylate cyclase polypeptide (PACAP) and its receptor PAC1 in modulating the nodose ganglion of the vagus nerve to NTS and the NAmb to heart circuits, shedding light on cellular mechanisms of interoceptive signaling during stress. The Maximizing Investigators' Research Award (MIRA) R35 project will elucidate the intricate neural mechanisms connecting the brain and heart during stress, focusing on two communication mechanisms: bottom-up (nodose ganglion to NTS) and top-down (NAmb to heart). The current project fits well with NIGMS's focus on multiorgan physiology under stress in the Division of Pharmacology, Physiology, and Biological Chemistry. In this proposal, we will explore the impact of the nodosePACAP to NTS and NAmbPACAP to heart on behavior, cardiorespiratory, and cardiovascular functions. The MIRA for Early-Stage Investigators (ESI) allows simultaneously investigating the brain and heart communication in stress through complementary top-down and bottom-up pathway analyses. In the next five years, our research will uncover stress-related organ-organ communication mechanisms, focusing on neuropeptide pathways and understanding the intricate processing of bodily signals. The project's overarching vision is to rigorously investigate stress-related effects on brain-body communication, addressing a critical gap in stress biology research. While rooted in basic science in mice, the research has broader implications for practical applications to general medicine.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Type 2 inflammation is mediated by a specific branch of the immune system, assumed to have developed as a major defense mechanism against parasitic infections and infestations. However, in our modern lifestyle, this kind of immune axis is mostly encountered in the context of pathological activation, especially in inflammatory diseases such as atopic dermatitis (AD). Consistently, the type 2 blocking antibody dupilumab was the first targeted FDA-approved treatment for AD. However, only 50% of patients achieved a 75% improvement in their skin scores upon dupilumab treatment during phase III clinical trials, and 10% even develop de novo paradoxical inflammatory side effects such as blepharitis, conjunctivitis, psoriasiform rashes, or a specific form of dupilumab- associated head/neck dermatitis (DAHND). Until today, mechanisms underlying treatment resistance and paradoxical inflammatory side effects of dupilumab treatment remain only insufficiently understood. Thus, there is an urgent need to correct this knowledge gap for the development of better treatment modalities for AD. Preliminary data from our group show increases in type 1 and type 22 immune signatures in dupilumab non- responders at baseline and throughout treatment. In addition, we find oligoclonal expansion of cytotoxic type 22 T cells in biopsies of DAHND. Interestingly, these cells showed overlapping features with type 2 cells, suggesting a considerable degree of inflammatory T cell plasticity, or transdifferentiation, both in dupilumab-resistant AD and in DAHND. In this proposal, we will utilize human skin biopsies from dupilumab-treated AD patients to study cellular and molecular features of the immune microenvironment in resolving (dupilumab responders), stable (dupilumab non-responders) and paradoxically flaring skin lesions (DAHND and psoriasiform rashes) through high-throughput techniques including single-cell RNA-seq, single-cell ATAC, and spatial transcriptomics. T cells isolated from these lesions will be studied in in vitro cultures and in skin spheroid models to better characterize their cytokine production behavior over time, as well as their crosstalk with cells of the tissue microenvironment, respectively. The rationale for this proposal is that once it is understood how type 2 cells are regulated in AD during individual treatment responses, these mechanisms can be utilized to create effective and novel stratified therapies for AD.

NIH Research Projects · FY 2025 · 2025-09

Project summary Chronic HIV infection produces pathologic inflammation which drives disease progression and contributes to the development of serious co-morbid medical conditions, even in the setting of effective combination antiretroviral therapy (CART); translocation of bacterial products across the gastrointestinal (GI) mucosa is a major antigenic stimulus for this process. Our research focuses on how HIV-associated autonomic neuropathy (HIV-AN), which is part of the spectrum of HIV-associated neuropathies, affects GI and immune function in people with HIV (PWH). This application describes a continuation of work begun as part of R01DK122853 “Effects of Vagal Dysfunction on Gastrointestinal and Inflammatory Pathways in HIV” (2/11/2020-01/31/2025). The title of this new submission is: “Complex Autonomic Neuropathic Dysfunction Leading to Gastrointestinal and Immune Effects in People with HIV (CANDLE),” which reflects a shift away from vagal dysfunction toward a more holistic consideration of the autonomic nervous system (ANS). In the prior grant we found that: 23% of PWH experience prolonged small bowel transit time (SBTT) as assessed using a Wireless Motility Capsule (WMC); prolonged SBTT is associated with small intestinal bacterial overgrowth (SIBO); HIV-AN is associated with alterations in immune function, including elevations in IL-6; HIV-AN independently predicts morbidity and mortality; and HIV-AN has heterogenous phenotypes with variable dysfunction in different branches of the ANS. Operationally, the new project will have two components: 1) longitudinal study of 125 PWH and 50 HIV- negative controls; 2) in depth, multidimensional analysis of samples/data collected during the prior R01 which include gold standard autonomic function testing; time-stamped, continuous whole gut pressure data from the WMC; saliva and stool samples for microbiome analyses; and blood samples for analysis of immune cells (using CyTOF and scRNA-seq) and plasma immune biomarkers. Our overarching goals will be to solidify the etiologic role for HIV in the development of HIV-AN, and to understand the time course of HIV-AN development and progression; and to continue to build on our understanding of the GI-mediated and direct pathways between HIV-AN and systemic inflammation which was begun in the prior R01.

NIH Research Projects · FY 2025 · 2025-09

Skin injuries and repair failures are a significant public health concern, with over 100 million new acute skin wounds occurring annually and non-healing wounds affecting 2% of the U.S. population. The economic burden of wound management is substantial, with annual treatment costs exceeding $25 billion. Despite this impact, effective interventions to improve skin healing outcomes remain elusive. Skin injuries expose internal organs to environmental threats and allow the translocation of surface microbes. While the pathological role of microbes in non- healing wounds is well studied, their contribution to acute healing and the potential of specific microbial factors to serve as pro-repair signals remain unclear. We and others have shown that signals from the skin microbiome can modulate cutaneous immunity and barrier function, suggesting that microbially-derived signals may also enhance repair. Our strong preliminary data show that several Staphylococcal species are enriched in acute human wounds. Using an in vivo screen with a newly developed physiological microbial translocation wound model, we identified specific Staphylococcal species that enhance repair four-fold without causing infection. Loss-of-function bacterial genetics revealed that enhanced Staphylococcal-driven healing depends upon toxin production. We hypothesize that Staphylococcal toxins act as novel pathogen-associated molecular patterns for repair by modulating both epidermal and dermal repair processes. Aim 1 will identify the specific toxins and epithelial responses involved in promoting repair, while Aim 2 will investigate how microbial toxins drive dermal repair responses. This research will leverage cutting-edge spatial omics technologies, including spatial transcriptomics and spatial lipidomics, combined with rigorous bacterial genetics and in vivo functional studies in repair. Our efforts promise to uncover non- canonical microbial mechanisms that enhance skin repair, potentially leading to innovative microbe-based therapies to boost this fundamental process and increase barrier fitness.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Caspase activation platforms (CAPs) are multi-oligomeric complexes essential for initiating cell death or inflammation through the activation of initiator caspases. CAPs are pivotal players in these processes; they are responsible for both sensing lethal/pathogenic stresses and, in turn, engaging caspase activity in the cell. CAPs have come to define entire signaling pathways and include the apoptosome (cyt-c/APAF1/caspase-9), DISCs (DR/FADD/caspase-8), inflammasomes (NLR,AIM2/ASC/caspase-1) and the CAP for caspase-2 (C2), the PIDDosome (PIDD1/RAIDD/C2; the main focus of our lab for the past decade). Recent evidence points to the existence to at least one additional CAP for caspase-2 which, in contrast to the DNA damage-sensing, nucleolar PIDDosome, assembles in the cytoplasm in response to cytoskeletal injury. This RAIDD-dependent but PIDD1-independent C2 CAP may also be responsible for C2 activation and ensuing neuronal cell death in response to beta-amyloid (Ab), as suggested by elegant experiments in an in vitro model of Alzheimer’s disease (Carol Troy, Columbia University, collaborator). However, the molecular identity of this novel C2 CAP distinct from the PIDDosome has remained unknown. Through a genetic screen and additional preliminary data, we identify a strong candidate for this complex, in which the cytoskeletal adaptor, ankyrin-G (ANKG), substitutes for PIDD1 as scaffold, thus physically coupling cytoskeletal integrity to C2. In ongoing experiments, the newly discovered ANKG/RAIDD/C2 complex, which we designate ‘ANKosome’, assembles and drives cell death in mammalian cells exposed to cytoskeleton-targeted anticancer drugs. In this R21 application, the first to study the ANKosome, we will identify the precise ANKG isoform which nucleates the complex and, through structure/function analyses, elucidate the mechanism by which cytoskeletal injury triggers ANKosome assembly via ANKG. We will also delineate the biologic function(s) of the ANKosome, with two main hypotheses. First, we will test whether the complex truly transduces cytoskeletal damage into an apoptotic response in both mammalian cells and live zebrafish embryos. Second, we will test whether the ANKosome mediates apoptosis in murine neurons exposed to Ab using the Troy lab’s Alzheimer’s disease model discussed above. In further support of this latter hypothesis, high levels of ANKG have been reported to accumulate at Ab plaques in Alzheimer’s disease, contributing to the neurodegenerative phenotype. This short term, high risk/reward project seeks to lay the foundations for a new field in apoptotic signaling, that of the ANKosome, with potentially broad impact ranging from AD to cancer treatment, that is, two of the major public health challenges facing our aging population.

NIH Research Projects · FY 2025 · 2025-09

The Cardiovascular Science Training Program (CSTP) at the Icahn School of Medicine at Mount Sinai School (ISMMS) is dedicated to recruiting and training a cohort of pre-doctoral and post-doctoral scientists, aiming to foster their independent careers in cardiovascular research. Given the persistent health challenges associated with cardiovascular disease, understanding the cellular and molecular foundations of cardiac pathologies is crucial for discovering effective treatment strategies. The Icahn School of Medicine is ranked No. 10 in the nation for NIH funding by the Blue Ridge Institute for Medical Research, as well as Number 1 in New York and No. 4 globally on Newsweek’s list of “The World’s Best Specialized Hospitals.” Drs. Filip Swirski and Carol Gregorio, the Program Directors, are internationally-recognized scientists with outstanding credentials in leadership, research and mentorship. The training program offers its trainees vast opportunities in different areas relevant to cardiovascular science, including but not limited to Vascular Biology, Cardioimmunology, Electrophysiology, Myocardial Disease, Genetics, Development, Metabolic Diseases, Stems Cell/Gene Therapeutics, Artificial Intelligence, Systems Biology, and Drug Discovery, with cross- cutting opportunities in disciplines that highlight programs at Mount Sinai that are well-developed and internationally-recognized including advanced imaging and genomics/personalized medicine. The Cardiovascular Research Institute (CVRI), under the leadership of Dr. Swirski, along with various translational research institutes on the ISMMS campus, provides trainees with access to cutting-edge research facilities. The CSTP offers training positions to graduate students, Ph.D. post-doctoral fellows, and physician-scientist fellows, each with specific selection processes tailored to their respective educational backgrounds. The program offers intensive fellowships in one of 27 laboratories, didactic training, professional development, as well as rigorous training in the responsible conduct of research and in methods for enhancing reproducibility. Trainees participate in weekly research seminars and have access to courses through the Mount Sinai Graduate School of Biomedical Sciences. Annual evaluations, based on well-defined criteria for professional advancement, assess trainee progress. Opportunities for interaction with established investigators and past trainees, who have successfully advanced through the CSTP, are provided. Each trainee, in collaboration with their primary mentor, develops an individual development plan evaluated by the Program Directors. Internal and external advisory committees evaluate the program based on benchmarks such as trainee progress, recruitment success, extramural funding, and the career paths of former trainees. Our primary objective is to cultivate the next generation of fundamental and translational investigators, including clinician/scientists, within an environment that nurtures their professional growth and independence.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Alzheimer's disease (AD) is commonly associated with social behavioral abnormalities and neuropsychiatric symptoms (NPS). In fact, NPS are increasingly recognized as core features of AD. However, research in AD has been primarily focused on the diagnostic and therapeutic treatment of memory deficits. There is an urgent need to better understand the neurobiology, behavior, and underlying mechanisms of NPS in AD. In response to PAR-23-207, we propose to investigate the influences of individual vulnerability to stress on the expression of NPS syndromes in mouse models of AD and to test the hypothesis that promoting stress resilience could prevent or delay the future emergence of AD-associated NPS. Stress is a major risk factor for neuropsychiatric disorders. In mice, we found that stress-susceptible (SUS) mice exhibit a spectrum of social, motivational, and reward deficits that are similar to the NPS, such as apathy, anxiety, and social withdrawal in human AD. We established that stress-susceptibility is associated with aberrant microglia activation in the prefrontal cortex (PFC) and, in turn, disrupts PFC neurophysiological properties. Moreover, gene expression analysis in the PFC of SUS mice showed several differentially expressed genes resembling those of disease-associated microglia (DAM) found in AD. Critically, stress- resilient mice maintain healthy-like PFC neurophysiological properties and healthy-like microglia activity. Our data shows that by manipulating microglia activity during stress exposure, we prevented stress-induced anhedonia and social withdrawal. Based on these observations, we hypothesize that vulnerability to prior stress exposure, stress-SUS, may prime PFC microglia in the PFC and promote the initiation and progression of AD-associated NPS behavioral and neuronal deficits, and oppositely, the active resilient mechanisms of stress-RES individuals, could prevent or delay AD-associated NPS and neuronal deficits. Using the 5xFAD and the 3xTg mouse models and leveraging two different stress paradigms: chronic social defeat stress and chronic variable stress, we propose to promote stress-resilience and to assess its short-term and long-term impacts on 1) NPS-like behavior development; 2) cortical neuronal activity and molecular underpinnings, and 3) circuit-connectivity associated with NPS-like phenotypes. To account for sex as a biological variable, we will characterize molecular, electrophysiological, and behavioral responses to stress, and AD-type NPS predispositions in female and male mice. We will employ cutting-edge methodology from cognitive and affective neuroscience and integrate multiple levels of analysis to address neurobiological, behavioral, and molecular mechanisms. These studies will uncover the active resilient mechanism(s) in the brain and offer novel targets for the prevention and treatment of NPS in the context of AD.

NIH Research Projects · FY 2026 · 2025-09

Project Summary The United States is in the midst of a significant paradigm shift around the perception and use of cannabis. The majority of states now permit recreational or medical use, reflecting a shift in views of cannabis with many believing the drug is not harmful or addictive. Despite the rise in these beliefs, repeated large-scale reports show that 10-30% of regular users meet the criteria for cannabis use disorder (CUD), an addictive disorder characterized by loss of control over use and relapse. As cannabis becomes more broadly available, it is essential that we determine the foundational mechanisms that drive problematic use. A major obstacle in studying CUD has been the lack of preclinical models of drug use. To bridge this gap, we have implemented a translational model of volitional edible ∆-9-tetrahydrocannabinol (THC; the main psychoactive constituent in cannabis) consumption to assess drug taking and seeking. With this paradigm we observed that unlike intravenous self-administration models, rats readily consume THC gelatin. Critically, this model recapitulates drug taking and cognitive phenotypes observed in CUD including increased drug intake and cue-induced seeking in males as well as individual differences in the capacity to titrate intake. This model provides a powerful foundation to assess neurobiological mechanisms that drive CUD-like behavior. Such clear sex and individual differences suggest potential involvement of epigenetic factors contributing to these phenotypes. Drugs of misuse like cannabis induce changes in epigenetic mechanisms such as histone modifications (e.g. methylation) which impact the accessibility of chromatin, permitting or suppressing gene expression. These mechanisms can activate or inhibit molecular machinery underlying synaptic plasticity leading to addiction-like behaviors. Recent clinical data indicate cannabis use is associated with changes in DNA methylation, and animal models have demonstrated a causal impact of THC exposure on epigenetic substrates throughout the mesocorticolimbic system. Using unbiased RNA sequencing of the nucleus accumbens core (NAcC), a reward subregion implicated in addiction, we observed dysregulation of transcripts underlying synaptic plasticity associated with changes in histone methylation. Although epigenetic mechanisms are a promising target for mitigating addictive behaviors, these mechanisms are tightly coordinated in a cell-type specific manner which can influence phenotypes. To improve the development and translation of novel therapeutics for addictions like CUD that target epigenetic mechanisms, it is essential we develop a cell-type specific understanding of the recruitment and role of these mechanisms in problematic cannabis use. This award will be used to interrogate the impact of histone methylation of distinct NAcC neuron outputs on the transcriptome, calcium activity, and regional networks. This program will be the first to show the causal impact of epigenetic modifiers on behaviors that promote and maintain CUD and will provide critical insight into novel therapeutic targets.

NIH Research Projects · FY 2026 · 2025-09

ABSTRACT Alzheimer’s Disease (AD) and AD-related dementias (ADRD) are devastating neurodegenerative disorders characterized by progressive cognitive decline and neuroinflammation. While epigenetic and protein-coding gene alterations have been widely studied in relation to brain aging, the role of RNA modifications, particularly N6-methyladenosine (m6A), remains less understood. m6A modifications influence gene expression, RNA stability, and cellular responses, but their specific contribution to AD/ADRD pathogenesis is largely unexplored. Microglia, the brain’s resident immune cells, play a central role in AD, contributing to processes such as amyloid-beta clearance and synaptic remodeling. Many AD-associated genetic risk loci, including TREM2 and APOE, are highly expressed in microglia, and microglia-specific regulatory regions capture a significant proportion of AD heritability. However, the post-transcriptional mechanisms driving microglial responses in AD, particularly through m6A RNA modifications, are poorly understood. Given microglia's critical role in neuroinflammation and AD pathogenesis, understanding how RNA modifications in these cells contribute to disease could reveal novel therapeutic targets. We hypothesize that dysregulation of m6A RNA modifications in microglia plays a key role in AD/ADRD pathogenesis by altering gene expression, disrupting cellular homeostasis, and exacerbating neuroinflammation. To investigate this, we will systematically map m6A RNA modification landscapes in microglia isolated from human brain tissue and human induced pluripotent stem cell (hiPSC)-derived microglia models using advanced long-read RNA sequencing. We will also use CRISPR-based perturbation of m6A writers, erasers, and readers to assess the functional consequences of these RNA modifications on microglial activities such as phagocytosis, cytokine production, and inflammatory responses. Aim 1 will profile m6A RNA modifications in microglia from AD and control brain tissues using long-read RNA sequencing, identifying differential m6A sites and investigating their interactions with AD risk loci to understand how these modifications influence microglial function. Aim 2 will utilize CRISPR-based approaches to perturb key m6A regulatory proteins in hiPSC-derived microglia and assess the impact on functions relevant to AD pathology. By integrating cutting-edge genomic technologies with functional assays, this study will provide crucial insights into how m6A RNA modifications contribute to AD-related neuroinflammation and microglial dysfunction. Ultimately, this research has the potential to uncover novel therapeutic targets, advancing efforts to develop RNA-based interventions for AD/ADRD.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY This is a resubmission of an application for a K23 Mentored Patient-Oriented Research Award by Dr. Francesca Cossarini, an Assistant Professor of Infectious Diseases at the Icahn School of Medicine at Mount Sinai and a current KL2 Scholar at the same Institution. This application aims to investigate the effects of proposed HIV cure strategies in the gut-associated lymphoid tissue. Despite the remarkable progress in the survival and quality of life of people living with HIV, forty years from its first discovery, HIV infection remains uncurable due to the persistence of a small pool of cells harboring the virus that can replicate if antiretroviral treatment is stopped. These cells constitute the HIV viral reservoir and efforts to identify and eliminate these cells have been ongoing for the past decades. Most of these studies so far have evaluated the effects of cure strategies in the peripheral circulation. With this proposal, I aim to evaluate the gut-specific effects of proposed cure strategies. The overall objective of this proposal is highly significant since a large proportion of the HIV viral reservoir is located in the gut-associated lymphoid tissue, and there is evidence of cross-infection with the peripheral circulation. Additionally, although memory CD4+ T represent the vast majority (if not the entirety) of the viral reservoir in the circulation, other cells likely function as reservoirs in intestinal tissue, however, their response to HIV cure strategies has not been evaluated. Dr. Cossarini is establishing herself as a physician-scientist in the field of HIV pathogenesis, remission, and cure. She has a specific interest in the role of gut-associated lymphoid tissue in the establishment and maintenance of the HIV reservoir, an area that has received less attention in the field compared to systemic circulation but is nonetheless critical for any strategy looking at potential cure strategies. The results of this study will contribute to addressing a gap in knowledge in the field of HIV cure strategies and will allow the PI to acquire robust data that will be tested in future research applications. With the completion of this proposal, the PI will also be able to further her knowledge in the field and master the analytical and laboratory skills to successfully transition to independence.

NIH Research Projects · FY 2025 · 2025-08

I am a dually trained neuroscientist and environmental epidemiologist with a primary research interest in understanding how early-life environmental exposures shape brain development. This R00 proposal supports my transition to an independent, transdisciplinary investigator in the emerging field of big-data environmental developmental neuroscience. Leveraging rich, existing behavioral and multi-modal MRI data from the Healthy Brain Network (HBN), a large-scale neurodevelopmental biobank, I propose to investigate how prenatal exposure to fine particulate air pollution (PM2.s) influences neurodevelopmental outcomes. Using a novel spatiotemporal model, I will estimate weekly PM2.s exposure throughout gestation and apply lagged weighted quantile sum (LWQS) regression to identify sensitive windows of exposure. Rather than relying on diagnostic categories, I adopt a cross-diagnostic, dimensional approach that focuses on continuous measures of social and attentional functioning-key domains impacted in both autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD). In this R00 phase, I will acquire longitudinal behavioral and neuroimaging data to examine how prenatal PM2.5 exposure shapes developmental trajectories in these domains. This project advances our understanding of the neurobiological mechanisms linking environmental exposures to neurodevelopmental disorders and provides a foundation for preventive strategies and targeted interventions. My long-term goal is to establish an independent research program using large-scale data to uncover environmental determinants of brain and behavioral development in children.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract This Mentored Clinical Scientist Research Career Development Award (K23) is designed to prepare Katherine Guttmann, MD, MBE, Assistant Professor and Attending Physician at the Icahn School of Medicine at Mount Sinai (ISMMS) and the Mount Sinai Hospital to achieve her career goal of becoming an independent researcher focused on communication between parents of infants and providers in the Neonatal Intensive Care Unit (NICU) or Pediatric Intensive Care Unit (PICU). She is driven by the overarching aim of improving communication between parents of critically ill infants and clinicians to enhance decision-making and optimize outcomes for children and families. In the short term, she seeks to secure the mentorship, training, and experience to successfully compete for R01 grants designed to improve communication between parents of critically ill infants and providers in order to improve parent and patient outcomes. To achieve these goals and transition to independence, Dr. Guttmann and her mentors have developed a detailed career development plan comprised of: (1) mentorship from a team with whom she has collaborated and published impactful work; (2) advanced training in family-centered communication, outcomes research methods, and implementation science; (3) mentored leadership training, and (4) an innovative research plan to define high quality communication with parents of critically ill infants, validate a measure of communication quality for such parents, and explore the impact of poor quality communication in the ICU. Intensive Care Unit admission creates psychosocial morbidity for parents of critically ill infants. Extrapolating from studies in adult contexts, the long-term impact of poor communication in the NICU or PICU is likely lasting and negative yet the depth and breadth of the impact of communication quality on parents and infants is not known. Communication in pediatric settings serves unique functions yet it is incompletely understood and there is not a high quality validated measure of communication quality specific to critically ill infants with parents of critically ill infants; 2) evaluating the psychometric properties and validity of a communication quality measure in parents of critically ill infants; and 3) exploring the long-term impact of ICU communication on parents. The proposed K23 will address a fundamentally important gap: lack of knowledge regarding quality of communication with parents of critically ill infants and its impact on families. The findings from this work will directly inform future R01 proposals which seek to improve communication in order to improve patient and family outcomes. career development plan delineates a carefully constructed approach to acquiring the skillset, expertise, and experience needed to become an independent researcher and a leader in communication quality research.

NIH Research Projects · FY 2025 · 2025-08

Summary All forms of diabetes result from insufficient numbers of insulin-producing beta cells. Thus, beta cell regenerative drug therapies could provide a scalable and affordable approach for millions of people with diabetes. The most advanced beta cell regenerative therapies include inhibitors of the kinase DYRK1A, exemplified by harmine, alone, or with GLP1 receptor agonists (GLPRA’s). However, the complete mechanisms of action for DYRK1A inhibitors and GLP1RA’s are incompletely understood. Therefore, to further optimize their efficacy, and to more deeply understand the mechanisms of action of beta cell regenerative therapies, a deeper cellular and molecular analysis of regenerative drug-treated human islets is needed. My Preliminary Data from single studies on regenerative drug-treated human islets suggest that “cycling alpha cells” serve as a beta cell progenitor reservoir in response to DYRK1A inhibitors such as harmine. However, most of our studies and reports from others have focused on beta cell proliferation at one single time point. As it is clear that islet cell subpopulations may progress through different phenotypes and proliferation phases over time following drug exposure, the mechanisms through which beta cell expansion occurs over time with regenerative drug treatment remain largely unexplored. Therefore, in this application I will address this knowledge gap by utilizing a human islet-derived organoid (islet MT) system that allows for long-term culturing and drug treatment. This will allow accurate study of lineage trajectory dynamics through collection of data at multiple time points. More specifically, in Aim 1 of this project, I will perform single cell and single nucleus RNAseq as well as single nuclear ATACseq, to explore temporal evolution of cellular mechanisms through which harmine enhances human beta cell regeneration in vitro. These experiments will also provide deeper insights into the gene regulatory networks that regulate transcriptional as well as epigenetic control. Since MT organoids are not identical to normal human islets, it is critical to compare data from Aim 1 with my existing single cell transcriptomic profiles of drug-treated human islets. Therefore, In Aim 2, I will integrate the single cell transcriptomic and accessible chromatin profiles of the human islet organoids from Aim 1 and compare and contrast these data with the single cell transcriptomic profile of human islets. Collectively, these analyses will be instrumental to pinpoint the temporal control of the fate decisions of cycling alpha cells and elucidate other possible islet cell types with the potential to transdifferentiate into beta cell.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY Mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene are the primary cause of cystic fibrosis (CF) and are associated with many other genetic diseases. Thus, CFTR is a compelling therapeutic target for treating various CFTR-related disorders. Genome editing techniques can correct the defective genes at their native locus and permanently restore the gene function, potentially offering a one-time curative treatment for CF. However, given that over 1,700 CF-causing mutations have been identified in the CFTR gene, mutation- specific genome editing methods, such as base editing, may not be suitable for correcting the wide range of CFTR mutations. To address these limitations, we propose to develop a more generalizable genome editing strategy for CFTR correction. Because most CFTR mutations occur within the exons or at the exon-intron boundary, we propose to replace the frequently mutated exons with their wild-type counterpart. Therefore, a single set of gene editing agents can correct all the mutations within the same exon. Prime editing (PE) employs an extended guide RNA (pegRNA) as a template for a conjugated reverse transcriptase to incorporate desired edits into the targeted genomic site. Recently, we devised a paired prime editing strategy to program the replacement of target genomic sequences without requiring a DNA donor in vitro and in vivo. Building upon this work, the goal of this proposal is to develop paired prime editing-based sequence replacement strategies to correct a broad spectrum of CFTR mutations, regardless of the mutation types or locations. We hypothesize that replacing the defective CFTR exons with corrected ones will effectively restore CFTR protein expression and function in vitro and in vivo. In the proposed research, we will utilize a donor-free sequence replacement strategy to replace individual CFTR exons without creating double-stranded DNA breaks or requiring a DNA donor (Aim 1). To further expand the editing scope and efficiency of the paired prime editing approaches in inserting or replacing large DNA sequences, we will develop a paired PE-based homology-independent insertion strategy and apply it to correct CFTR mutations caused by large deletions or occurred in the large exons (Aim 2). Finally, we will explore the therapeutic applications of the newly-established paired prime editing-mediated gene correction strategy by delivering the genome editing agents to the lungs of a CF mouse model (Aim 3). Successful completion of this project will result in a gene therapy strategy tailored for CF and provide a universal gene correction framework for treating genetic diseases of high allelic heterogeneity.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY I am an early-stage clinician investigator and an advanced multi-modality imaging cardiologist at the Icahn School of Medicine at Mount Sinai. My goal is to become a leader at the intersection of aging, cardiovascular imaging, and cardiovascular disease. My work focuses on how best to deliver patient-centered care and improve outcomes of older adults through use of cardiovascular imaging for stable ischemic heart disease (SIHD). Assessment of frailty is one key tool to improve these outcomes, as one in four of the 10 million older adults in the U.S. with SIHD is frail. Frailty is associated with increased risk of complications, hospitalizations, health care costs and mortality among patients with or without SIHD. Unlike most geriatric syndromes, frailty can be reversed with targeted interventions if diagnosed early and can guide personalized treatment decisions. Yet, frailty is rarely assessed in clinical practice due limited time and resources, or unfamiliarity with screening tools. This may lead to clinicians withholding appropriate beneficial treatment options for certain patients or subject other patients to inappropriate treatments that increase harm with limited benefit. Novel frailty screening tools that overcome these barriers are critical to increase frailty screening in clinical practice. The opportunistic use of existing imaging data acquired for other clinical indications is an innovative application for frailty measurement. These imaging data are ubiquitous: majority of the more than 1.5 million stress nuclear myocardial perfusion imaging (MPI) studies performed annually in older adults for diagnosis and management of SIHD also acquire low-dose chest CT images to improve image quality and diagnostic performance. In this project, I will develop and validate a novel chest CT-based frailty screening tool using artificial intelligence (AI) assisted multi-organ body composition information available from existing chest CT data performed with stress MPI. Next, we will compare the prognostic performance of the new CT-based tool with a validated electronic health record-based electronic frailty index to predict healthcare utilization and death. Finally, we will adapt this screening tool for integration within routine clinical workflow, and pilot test its feasibility using a pre-post approach among older adults presenting to an outpatient cardiology clinic visit for clinical decision making after stress imaging. The results of our study will integrate novel frailty screening into routine cardiovascular practice, to help guide patient-centered treatment decisions, target effective prevention strategies, and inform my future R01, which will test AI-enhanced patient evaluation strategies among older adults. This Beeson award will support my training in (1) geriatrics and geriatric cardiology; (2) clinical applications of AI (3) dissemination and implementation science and (4) leadership. Our approach can expand the availability of real-time point-of-care frailty screening without additional clinical burden by using existing CT data. The Beeson award will help me achieve my goal to become a leader in aging who develops novel strategies aligning imaging use with the distinct needs of older adults with SIHD.