Icahn School Of Medicine At Mount Sinai

NSF Awards · FY 2026 · 2026-07

The immune system protects the body by recognizing abnormal cells and infectious threats, yet the molecular rules that allow immune cells to distinguish dangerous targets from healthy cells remain poorly understood. This project will develop new computational approaches to better understand immune recognition, a fundamental problem in biology with broad relevance to health, cancer immunotherapy, autoimmunity, and future biomedical discovery. The project will also advance computing by developing new methods to model biological systems using structure-aware protein language models. In addition to the research activities, the project will support a month-long summer program in New York City that introduces high school students to immunology, data science, and machine learning through hands-on projects and mentorship. Openly shared software, educational materials, and datasets will help broaden access to computational biology and strengthen the future scientific workforce. This project will develop structure-aware protein language models to decode how T cell receptors recognize peptide antigens presented by major histocompatibility complex molecules. The research will generate high-confidence three-dimensional models of paired T cell receptors, represent local structural environments as symbolic tokens, and integrate sequence, structure, and spatial information in a transformer architecture trained with masked multimodal prediction and contrastive learning. The resulting representations will be tested for their ability to identify shared antigen specificity across diverse immune repertoires and will then be integrated with single-cell transcriptomic profiles to link receptor recognition with cellular function. Experimental studies will be used to evaluate model predictions and iteratively refine the framework. The project is expected to produce new computational methods for modeling molecular recognition, new mechanistic insight into immune specificity, and open tools that can be reused across immunology and machine learning–driven biology. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Lung possesses a remarkable ability to regenerate and restore normal lung function after severe acute injury. However, in the case of chronic injuries, the lung epithelial stem/progenitors fail to restore normal lung function and contribute to a pathologic regeneration, amounting to regenerative failure that typifies chronic restrictive lung diseases like Idiopathic Pulmonary Fibrosis (IPF). Both airway and alveolar epithelial stem cells are capable of differentiating into type 1 and type 2 alveolar epithelial cells (AT1 and AT2, respectively) via a series of intermediate cell states. During normal injury resolution, like in acute injuries in human lungs and murine models, both airway and alveolar epithelial stem cells go through a euplastic intermediate cell (EIC) state to eventually differentiate into alveolar epithelial cells (AT1 and AT2s). However, in chronic lung diseases like IPF or animal models of chronic fibrotic injury, the stem cells progress through EICs and differentiate further into aberrant basal-like dysplastic intermediate cell (DIC) state that result maladaptive regeneration and honeycomb-like cystic structures. Currently, we lack understanding of the signaling mechanisms that control the balance between euplastic and dysplastic regenerative cell states. This is especially relevant in a histologically heterogeneous disease like IPF where the balance between euplastic and dysplastic repair is a key indicator of disease severity. To this end, we discovered that EICs and DICs are spatially distributed in relation to severity of remodeling and state of microenvironment niche signals. EICs are often found in areas of active but mild injury and are associated with mesenchymal cells that secrete pro-regenerative growth factors such as NRG1 that activated ErbB3/STAT3 signaling. On the other hand, DICs are associated with Collagenhigh/CTHRC1high pro-fibrotic mesenchymal cells found in actively remodeling regions. Furthermore, our preliminary data suggests that intermediate filaments such as cytokeratin 17 (KRT17) and Vimentin play an integral role in the switch from EIC to DIC. Therefore, we will test the hypothesis that spatially restricted microenvironment cues activation of distinct signaling pathways in activated epithelial progenitors to determine euplastic versus dysplastic regenerative trajectories in the following three aims: Aim 1: Define the role of NRG1/ErbB3 signaling in euplastic alveolar regeneration. Aim 2: Determine the role of TGFβ1/KRT17 signaling axis in dysplastic alveolar regeneration during chronic injury. Aim 3: Identify spatial cues required for euplastic to dysplastic regenerative switch. We will use newly established animal models of chronic injury, cell-specific in vivo conditional knockout of relevant candidate genes, in vitro co- culture techniques modeling human euplastic and dysplastic repair to test specific signaling components of NRG1/ErbB3 signaling and TGFβ1/KRT17 signaling. Finally, using a novel in silico approach, we will build an integrated mouse PF and human IPF spatiotemporal signaling map of progressive fibrosis. Together, successful completion of this proposal will build a comprehensive model of niche-influenced signaling crosstalk governing epithelial plasticity and reveal novel therapeutic targets for treating fibrotic disorders.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT My goal is to understand the mechanisms underlying pruritus. The ability to sense the environment is essential for survival, with the somatosensory nervous system playing a central role in detecting external stimuli. Despite its evolutionary relevance, the sensation of itch (pruritus) has been historically overlooked, even though it significantly impacts quality of life, particularly in chronic cases. Of note, pruritus affects 15% of the population, yet there are very few FDA-approved treatments, making it a major unmet medical need. Recent studies highlight the crucial interactions between the immune and sensory systems, especially the role of unmyelinated C-fibers in driving both inflammation and pruritus. While C-fiber hyperinnervation is a feature of pruritus, the mechanisms driving this nerve overgrowth and its role in pruritus pathogenesis remain largely unknown. The skin is the primary sensory organ for pruritus and the ecological niche for commensal microbiota. Using a novel TCR transgenic mouse model with T cells specific for Staphylococcus aureus (S. aureus), I previously demonstrated that T cell immunity to commensal S. aureus promotes skin sensory nerve regrowth after injury via the IL- 17A/IL-17RA axis. While host-microbiota interactions are essential for maintaining immune balance and skin homeostasis, disruption of this equilibrium can transform commensals such as S. aureus into pathogenic drivers of inflammation and tissue damage, as observed in psoriasis and atopic dermatitis. Might microbiota- induced cytokines contribute to these outcomes? Building on the neurotrophic effect of microbiota-induced IL- 17A, and considering the established role of type 2 cytokines (IL-3, IL-4, IL-13, IL-31) in itch via sensory nerve activation, my lab is investigating whether IL-17A promotes maladaptive neuronal growth during inflammation, leading to sensory hyperinnervation and pruritus. To test this, we have used two models of skin microbiota (S. aureus and Candida albicans) combined with two inflammatory itch models: IMQ-induced psoriatic itch and MC903-induced atopic dermatitis. We have found that mice in the pruritus group (microbiota + IMQ/MC903) showed both skin hyperinnervation and heightened itch, driven by the IL-17A/IL-17RA axis. Furthermore, snRNA-seq of dorsal root ganglia (DRG) neurons and microscopy revealed a transcriptional signature consistent with axonal growth, including upregulation of Atf3, a key regulator of nerve growth following injury Based on this, I hypothesize that microbiota-induced T cells control skin sensory hyperinnervation upon inflammation, consequently regulating pruritus via the ATF3-IL-17RA axis. To test this hypothesis, I will ask three interrelated but non-dependent aims: Aim 1) How do microbiota-induced T cells control hyperinnervation, Aim 2) To elucidate the transcriptional regulation of IL-17RA- dependent sensory hyperinnervation, and Aim 3) How does ATF3-IL-17RA axis regulate pruritus? This project seeks to establish a novel paradigm of microbiota-driven neuroimmune regulation of sensory neuron plasticity, focusing on hyperinnervation, and long-term effects on pruritus, very relevant to target inflammatory skin conditions.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY: Monkeypox virus (MPXV) poses an escalating global health threat, with recent outbreaks extending beyond endemic regions. Current treatments and vaccines offer limited protection, and a new clade I resurgence in 2024 highlights the urgent need to understand MPXV pathogenesis. Maternal mpox infections have been linked to fetal loss and neonatal death, yet mechanisms of vertical transmission remain unclear due to the lack of suitable small animal models. We developed a pregnancy model to study MPXV in mice and established a human placental tissue model that supports MPXV infection. In this application, we will study pregnant mice as well as a human villous explant system, both of which support MPXV infection to understand how gestational age and viral clade influence infection outcomes in mice and humans. We will then test the extent to which current and next-generation therapeutics, alone and in combination, can alter infection and pathogenesis caused by MPXV. Finally, we will map MPXV-infected cells and the timing of infection at the maternal–fetal interface in mice and human villous explants. Together, this work will provide critical insights into MPXV pathogenesis in pregnancy and guide development of targeted therapies for congenital mpox.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract: APOE4 and aging are the among the strongest risk factors for Alzheimer’s disease (AD). As the aging population continues to grow in the coming decades, novel approaches based on uncovering new AD pathobiology will be needed to meet the increased number of cases. An emerging concept with accumulating support is that AD pathobiology is not limited to dysfunction occurring within the CNS. These studies demonstrate that the activity of youth-associated proteins in young blood revitalizes function in aged tissues, including various processes within the brain. Using plasma transfer and parabiosis models, aged mice sharing young blood exhibit improved behavioral function, including hippocampus-dependent memory, and improved synaptic plasticity. Aged amyloid-bearing mice exhibit reduced AD pathology and improved synaptic integrity following exposure to young blood. Conversely, factors in the aged systemic environment drive aging phenotypes in the young healthy brain, including microgliosis and deficits in cognition, together arguing that blood-borne factors are relevant to neuroimmunological changes relevant to AD pathogenesis. While these studies argue that the systemic environment may represent a target for novel AD therapies, little is known about the drivers of these phenotypes or other critical modifiers of blood-CNS communication that regulate AD pathology. Our preliminary data provide strong support for the concept that the systemic environment differs according to APOE genotype and that sharing blood from opposing APOE genotypes differentially alters hippocampal function. We will address the role of both APOE4-associated and APOE3-associated mediators of blood-CNS neuroimmune communication in the healthy brain and in the context of AD pathology to clarify the link between common risk factors and AD pathomechanisms. We hypothesize that hippocampus-dependent behavioral and neuroimmunological function is regulated by communication between the periphery and brain in a manner that depends on APOE genotype. We will address this in several aims: (1) To examine deleterious APOE4-associated blood-borne mediators of changes in brain function at multiple levels of analysis (molecular, cellular, and behavioral); (2) to similarly characterize the impact of protective APOE3-associated blood-borne mediators on neuroimmunological and behavioral function; and (3) to examine the impact of APOE genotype on the efficacy of young blood-borne factors in revitalizing function in the aged brain and in the context of AD pathology. Our aims incorporate the use of sophisticated transcriptomic, behavioral, and morphometric approaches to comprehensively define cellular and pathological responses to APOE genotype in the systemic environment, potentially opening novel avenues for AD therapy development.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Mammalian skin is characterized by the formation of hair follicles, which exhibit polarized growth, and distinct distributions and patterns in different body regions. Hair follicle development occurs during embryonic and fetal life and is suppressed after birth under normal conditions. Initiation of hair follicle formation requires activity of the Wnt/b-catenin signaling pathway, while hair follicle polarity is established by non-canonical Wnt/Planar Cell Polarity (PCP) signaling. How these pathways are modulated to allow for correct hair follicle orientation and distribution across the body, and to prevent formation of new hair follicles after birth, is incompletely understood. This study will test the hypothesis, based on preliminary data, that the unrelated, secreted Wnt inhibitors DKK2 and SOSTDC1 are required for normal hair follicle orientation and distribution, and act during postnatal life to suppress formation of new hair follicles. Aim 1 will determine the mechanism by which DKK2 and SOSTDC1 regulate hair follicle polarity during embryogenesis. Aim 2 will delineate how these inhibitors control the patterning of tail hair follicles and sebaceous glands. Aim 3 will examine the mechanisms by which DKK2 and SOSTDC1 suppress hair follicle neogenesis in postnatal skin. Delineating these mechanisms has potential to identify new treatments for scarring hair loss disorders and novel therapeutic approaches to regenerate haired skin in cases of severe wounds or burns.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Combination antiretroviral treatment has transformed HIV (Human immunodeficiency virus) infection from a fatal disease to a manageable chronic condition. It requires, however, regular antiretroviral medication. Attempts to cure HIV fall short because of the existence of long-lived HIV reservoirs comprised of cells harboring non- productive proviruses. While HIV is integrated into the genome of these cells, no viral proteins are expressed, leaving the cells indistinguishable from uninfected cells, preventing clearance by antiviral treatments or the immune system. My proposal seeks to define the properties and features of nonproductively infected cells circulating in the blood or residing in lymphoid tissues. My project will leverage a dual-reporter virus which allows for the physical separation of cells nonproductively infected with HIV from those that are productively infected and uninfected cells. Additionally, we have worked to develop methods of culturing and infecting lymphoid cells derived from human tonsils to mimic infection in tissues. Utilizing these model systems, we will work to understand cellular and viral factors which influence nonproductive HIV infection. In Specific Aim 1, I will utilize spectral flow cytometry to phenotype the cell types present in our tonsil organoid model, delineating the cells that harbor non-productive proviruses. We will compare these results to those found in blood derived CD4+ T cells and measure the impact of other bystander cell types on the relative frequency of productive to non-productive infections. We will use RNA sequencing to compare the transcriptomic landscapes of nonproductively infected cells to those of productively infected cells. In Specific Aim 2, we will utilize established and novel techniques to identify viral characteristics which correlate with nonproductive HIV infection. We will use digital PCR technologies to measure proviral intactness, and parallel sequencing techniques to identify HIV integration sites. These experiments will collectively identify host and viral factors which play a role in HIV latency establishment in relevant primary human cells and organoid model systems. This research will be supervised by Dr. Viviana Simon, a recognized leader in the field of virology. I will additionally be aided by esteemed collaborators both at our institution and beyond, who will assist with technical aspects as well as my development as a budding researcher. My fellowship proposal will generate new insights into the non-productive HIV reservoir, provide opportunities to learn advanced techniques both on the bench and in data analysis, and prepare me for a successful career as an independent academic researcher and mentor.

NIH Research Projects · FY 2026 · 2026-06

Stroke is a major cause of death globally. Several studies found a higher stroke risk associated with fine particulate matter (PM2.5), but critical knowledge gaps persist, necessitating urgent investigation. We are uniquely positioned to investigate the associations of pathway-specific hemorrhagic and ischemic (i.e., large artery atherosclerosis, cardio aortic embolism, small artery occlusion, other, undetermined) strokes, with short- and long-term PM2.5 exposure and its composition. We will leverage novel artificial intelligence (AI) models to classify pathway-specific strokes using 13 years of electronic health records (EHR) from the Mount Sinai Health System (MSHS). This comprehensive health and covariates data includes clinical notes, imaging, labs, medications, and diagnoses. MSHS, the largest health care provider in the New York City metropolitan area, serves an urban and suburban population, including all demographic groups. Aim 1 will build Generative AI-based models to categorize pathway-specific stroke cases in the EHR. This model will be generalizable to multiple healthcare systems and improve stroke research, which often combines distinct subtypes into a single endpoint despite their differing pathophysiology. By connecting air pollution exposure to specific stroke pathways, we will provide evidence on the underlying biological mechanisms driving these associations. Aim 2 will assess the association of PM2.5 mass and composition with pathway-specific stroke. Unlike most studies that obtain PM2.5 component measurements from ground monitoring sites, we have developed innovative, highly spatiotemporally resolved models that significantly minimize the exposure measurement error and improve our ability to estimate exposure- response associations. We will use an exposure-mixture approach to estimate the combined associations of the chemical components and identify the components attributing most to each pathway-specific stroke risk. Finally, Although the EPA has lowered the annual PM2.5 standard from 12 to 9 μg/m3, this threshold is still markedly higher than the threshold recommended to protect human health (5 μg/m3). Our previous analyses show that PM2.5 exposure <9 μg/m3 is still associated with increased cardiovascular risk. We will assess whether the associations remain considering different annual PM2.5 standards. Aim 3 will Assess whether the association between PM2.5 and pathway-specific stroke varies by demographic and clinical characteristics. We will quantify exposure variations and evaluate the extent to which the associations between PM2.5 and stroke differ across income and financial hardship levels considering lower PM2.5 standards. In summary, this study will advance the use of administrative data for large-scale environmental health research. We will develop an AI model that can provide accurate stroke identification and classification for patients nationwide. Our findings will provide scientific evidence on the health effects of individual PM2.5 components to inform targeted regulations and air quality standards. Additionally, this study will help shape policies to reduce air pollution exposures and support prevention and treatment strategies to protect cerebrovascular health, with emphasis on susceptible groups.

NIH Research Projects · FY 2026 · 2026-06

SUMMARY Human milk provides nutrients and important non-nutritive factors for infants that promote growth, development, and protection from infection1,2. Therefore, the World Health Organization (WHO) recommends exclusive breastfeeding for 6 months, then combining breastfeeding with solid foods for 18 months3. Lactation disorders reduce breastfeeding rates, and negatively impact both mothers and children. In mothers, lactation disorders influence mood and maternal well-being, while in children, they affect cognitive and socio-emotional development4, and can cause malnutrition, hypernatremia, hypoglycemia, and death5. Moreover, breastfeeding rates are lower in some ethnic minorities, which may partially reflect poor access to lactation consultants and early initiation of infant formula. Lactation disorders that specifically impair milk production and secretion affect about ~40% of breastfeeding mothers, the major phenotypes including: (i) agalactia: complete absence of milk secretion following birth; and (ii) hypogalactia: insufficient volume for optimal infant nutrition6,7. While post-partum stress, obesity, diabetes, and socioeconomic considerations have been associated with hypogalactia, we and others have demonstrated that hypogalactia has an inherited maternal genetic component6,8,9,10. However, the genetic mechanisms responsible for human lactation disorders are mostly unknown and have not yet been extensively investigated. We hypothesize that variations in genes involved in human milk production and secretion underlie disorders of milk production and secretion, and that these variants and genes can be discovered by interrogating genomic and extensive health and metadata from women with lactation disorders cases compared to unaffected female controls. We therefore propose to conduct a comprehensive study on whole exome sequencing (WES) data of lactation disorders patients. We are uniquely positioned to perform the first such study with the largest lactation disorders cohort to date (1,382 patients and over 60,000 female controls), combining four major biobanks: Vanderbilt University’s BioVU11,12, Mount Sinai Hospital’s BioMe Biobank13,14, All of Us and UK Biobank15,16. We propose a rigorous pipeline combining various state-of-the-art with cutting-edge approaches developed by us and others to: (1) obtain a high-quality WES lactation disorders cohort by variant- and sample-level quality control (QC)17,18, annotations19, and impact predictions20-22; (2) perform computational case-control analyses for high impact variants23,24; (3) prioritize variants and genes by biological relatedness approaches25-27 and use a novel quad-culture organotypic mammary gland model to characterize the molecular pathology of high impact variants; and (4) perform phenome-wide association studies (PheWAS)28 and polygenic risk score (PRS) analyses29. We expect that our findings of human lactation disorders genetics will be vital for understanding the physiology and pathophysiology of human milk systems, directly informing maternal, perinatal, neonatal health decisions, and ultimately guiding precision medicine approaches to improve women’s health.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Fibrosis and fatty infiltration impair skeletal muscle's regenerative properties and function in traumatic injuries and chronic diseases, and thus there is an urgent need for innovative therapies to restore muscle structure and function. Such muscle degenerative hallmarks are driven by muscle-resident mesenchymal stem cells, also known as fibro-adipogenic progenitors (FAPs). While these cells typically contribute to muscle regeneration by supporting myogenesis through proliferation, secretion of pro-myogenic paracrine factors, and deposition of matrix before apoptotic clearance, they can become resistant to apoptosis in chronic injuries, before differentiating into myofibroblasts and adipocytes. However, extrinsic factors controlling FAPs’ response to muscle injury are not well understood. One such factor may be the biophysical cues during muscle healing. While matrix stiffness regulates mesenchymal stromal cell fate, how FAPs sense the changing matrix stiffness and drive their function in muscle regeneration remains unknown. The mechanism of sensation may be through PIEZO1, a mechanosensitive ion channel. PIEZO1 has been shown to be present in many cells across different tissues, including cells found within muscle, and responds to a range of mechanical cues, especially substrate stiffness. However, it has not yet been identified in FAPs, nor the role PIEZO1 plays on activation. This project will elucidate how FAPs sense the mechanical changes in their environment and downstream processes of PIEZO1 activation. To address this gap, the role of PIEZO1 in mechanosensation in FAPs during muscle regeneration in vivo will be identified in Aim 1. Following conditional knockout of PIEZO1 in FAPs, the muscles will then be injured with barium chloride injection. Muscles without PIEZO1 will show worsened muscle regeneration and increased pathology. Furthermore, matrix stiffening will be prevented following muscle injury to assess the role of mechanosensation in activating PIEZO1 in vivo. Here, muscles with impaired matrix stiffening will behave similarly to those without PIEZO1, showing signs of impaired muscle regeneration and increased pathology, further implicating the importance of PIEZO1 in matrix stiffness sensation and healthy muscle regeneration. Aim 2 seeks to isolate the mechanism of PIEZO1 activation in FAPs and identify downstream effects of activation. Using engineered hydrogels to match stiffnesses that are seen during muscle regeneration, I expect stiffer hydrogels to cause increased PIEZO1 activation. Furthermore, any mediators that are downstream of PIEZO1 will be identified, with a focus on mechanotransductive genes. This project will frame the research within a clinical context and provide a multi-disciplinary training in tissue engineering and stem cell biology to build my career as a future physician-scientist as well as providing therapeutic targets to prevent the development and progression of muscle degeneration following injury.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Rotator cuff (RC) tears are a leading cause of musculoskeletal impairment, affecting over 250,000 individuals annually in the United States. Despite surgical repair being the standard of care, failure rates remain alarmingly high, with up to 90% of cases experiencing re-tears or poor functional recovery. This failure is largely driven by irreversible muscle degeneration, including atrophy and fatty infiltration, which impair healing and compromise surgical outcomes. Current therapies primarily target the bone-tendon interface while neglecting muscle pathology, underscoring a critical need for regenerative strategies that directly address RC muscle degeneration. This proposal aims to develop a therapeutic approach leveraging lipid nanoparticles (LNPs) encapsulating WNT7a mRNA (W7a-LNP) to promote muscle regeneration and prevent degeneration following RC injury. WNT7a has been shown to increase muscle mass, enhance muscle stem cell (MuSC) expansion, and reduce fatty infiltration, but its recombinant protein form is limited by poor bioavailability and high production costs. W7a- LNP circumvents these limitations by enabling localized, sustained WNT7a production at the injury site, transforming muscle cells into `in vivo protein factories.' Our preliminary data demonstrate that W7a-LNP reduces fibro/adipogenic progenitor (FAP) adipogenesis and fatty infiltration in both in vitro and in vivo models. We will test the hypothesis that intramuscular delivery of W7a-LNP prevents and reverses RC muscle degeneration through three specific aims. Aim 1 will engineer and validate W7a-LNP as a targeted muscle regeneration platform by optimizing delivery, evaluating WNT7a expression kinetics, and assessing its effects on MuSC expansion, myofiber hypertrophy, and FAP adipogenesis in vitro and in vivo. Aim 2 will determine the efficacy of W7a-LNP in preventing RC muscle degeneration when administered at the time of injury using a clinically relevant delayed tendon repair model. Aim 3 will evaluate W7a-LNP's ability to reverse established muscle degeneration when delivered at later stages of injury, with or without mechanical loading, to assess potential synergistic effects. The impact of this research is the development of a scalable, translatable mRNA-LNP therapy that preserves and restores muscle quality in RC injuries. By addressing a critical gap in current treatment paradigms, this strategy has the potential to improve surgical outcomes, enhance functional recovery, and reduce the need for invasive salvage procedures, ultimately transforming the management of RC muscle degeneration.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY The primary factor for the dismal outcome of glioblastoma (GBM) is high inter- and intra-tumor heterogeneity. By using bulk RNA-seq, the Cancer Genome Atlas (TCGA) initiative provided robust gene expression-based identification of three GBM subtypes: Proneural (PN), Mesenchymal (MES), and Classical (CL). These molecular subtypes are not mutually exclusive and can co-exist within a single tumor but are important to comprehensively characterize because they represent ends of the spectrum of different molecular gradients that shape the GBM tumor microenvironment (TME). Hence, better understanding of the TME of different GBM subtypes is a critical first step towards improving patient stratification as well as identifying therapeutic vulnerabilities for different malignant cell subpopulation that make up mixed-subtype GBM tumors. Preliminary data in this application defines two distinct GBM MES subtypes, associated with different genetic drivers, TME cell compositions, and survival of patients. We propose to combine publicly available single-cell RNA-sequencing (scRNA-seq) data from 199 GBM patients with emerging high-resolution spatial transcriptomics technologies to dissect the TME cell composition, spatial tissue organization, cancer-intrinsic, and myeloid-driven immunosuppressive mechanisms underlying differences in survival between the two MES GBM subtypes (Aim 1). We will additionally develop a new machine learning approach for discovering immunomodulators of TME cell composition and combine it with existing computational tools to model cell-cell interactions across the integrated cohort, which will enable us to uncover new, context-specific therapeutic targets. In Aim 2, we will build and comprehensively immunophenotype genetically engineered mouse models (GEMMs) of the two new GBM MES subtypes to dissect the role of distinct genetic drivers on the GBM TME. The GEMMs will also provide a validation platform for the computationally discovered therapeutic targets with conserved function in human and mouse GBM. Hence, our study promises to uncover human-relevant cell-cell interactions and pathways that lead to an immunosuppressive TME in MES GBM and to identify new strategies for immunomodulatory treatment and patient stratification. Finally, this proposal will create an integrated human-murine, single-cell resolution resource that will provide an analysis and experimental platform for target discovery and pre-clinical validation for the two MES GBM subtypes.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY – Overall Application: Immunologic Trajectories of Peanut Desensitization (INROADS) Peanut allergy has tripled in prevalence in recent decades, affecting nearly 2% of adults and up to 5% of children in some US regions. Peanut allergy is typically lifelong, can be fatal, and impacts quality of life. Our group and others performed the studies resulting in the recent FDA approvals of a commercial peanut oral immunotherapy (OIT) and an anti-IgE monoclonal antibody, omalizumab. These therapies can raise the reaction threshold and provide safety from accidental exposures for approximately 67% of patients. Additionally, in a prior period of our AADCRC grant, we focused on children whose reaction threshold to peanut was higher than the children included in the registration trials of the expensive pharmaceutical products. We demonstrated that nearly half of the peanut allergic population could be treated safely and effectively with inexpensive, retail store-purchased, home-measured peanut. These findings are revolutionizing desensitization therapy for peanut allergy and provide options for our patients other than strict peanut avoidance. However, in clinical practice, there is no way to predict response to therapy, and there is an insufficient understanding of the mechanisms that are involved in the success, or lack thereof, from these therapies. This proposal, Immunologic Trajectories of Peanut Desensitization (INROADS), builds upon discoveries and techniques from our prior AADCRC and our additional studies to address the knowledge gaps of these desensitization therapies that will improve personalized care and provide mechanistic insights for better treatments. INROADS will address the overarching hypothesis that dynamic changes in circulating cellular subpopulations and molecular networks are mechanistically linked to peanut desensitization outcomes. Project 1, MICRO-TRACK, will use high-dimensional immune profiling, allergen- specific T cell assays, and in vitro stimulation models to identify predictive biomarkers and mechanisms of immunologic control of desensitization therapies. Project 2, SPADE, will complement this effort using transcriptomics and machine learning to identify signatures of desensitization and desensitization classifiers to provide mechanistic insights and clinical tools for treating individuals with peanut allergy. We will synergize Mount Sinai’s cutting-edge clinical therapeutics program via a Clinical Core (PATHWAYS) to provide clinical data and biosamples from oral food challenge tests performed before and following desensitization treatments. The two research projects, interactions with the Clinical Core, management of biosamples and data, and communications with NIAID and the AADCRC network will be coordinated and supported by an Administrative Core and a Data Stewardship Core. In addition to creating a rich resource of clinical, immune, and molecular data that will be made publicly available for the research community, our integrated program will advance personalized medicine, enrich mechanistic understandings, and potentially identify new therapeutic targets for peanut allergy that will be informative for allergy to any food.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT The goal of this project is to uncover the mechanisms driving brain metastasis occurrence in breast cancer models that are clinically less associated with brain metastasis at diagnosis. Metastasis stands as the utmost obstacle to Breast Cancer treatment. While both genomic and epigenetic increase in complexity during metastasis, bone has been identified as the primary site of metastasis relapse in luminal breast cancer. More recently, we began to better appreciate the complexity of the metastatic cascade during breast cancer progression. In fact, we realized that cancer cells can expand from various metastatic sites to initiate new lesions which contribute to accelerating disease progression. Despite these clinical observations, the mechanisms driving these secondary metastasis processes remain largely unexplored. More recently, we have identified a poorly studied neuron-associated ligand, Neuregulin 3 (NRG3), as a bone metastasis-related factor. NRG3 was induced in bone metastasis via epigenetic mechanisms and demonstrated to enhance the seeding potential of breast cancer cells. Surprisingly, we found that in the brain, the ecosystem surrounding secondary micrometastases derived from bone-reprogrammed cancer cells was also NRG3-enriched, arguing for key roles of NRG3 in the brain metastasis initiation process. The ability of luminal models to transition from “bone-only” to brain metastasis-initiating cells suggests that NRG3 could enhance the brain metastasis potential of cancer cells. As brain metastasis is associated with worse survival outcomes in bone metastasis patients, we hypothesize that the bone-mediated NRG3 promotes brain metastasis by remodeling the brain microenvironment and establishing a pro-survival niche. To test our hypothesis, we will (i) elucidate the role of neuron-derived NRG3 on CTC chemoattraction from bone to brain, (ii) investigate the mechanisms of the “NRG3 niche” establishment in bone metastasis-derived BrM, and (iii) assess the therapeutic potential of NRG3 inhibition using experimental and spontaneous preclinical models. Ultimately, this paradigm-shifting proposal will narrow down a large gap of knowledge on the metastasis-to-metastasis seeding process and establish a new foundation for brain metastasis prevention and treatment in patients with bone metastasis as the primary site of distant relapse.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract The predominant myosin heavy chain expressed in human heart, beta-MyHC, is encoded by the MYH7 gene. MYH7 variants are well described in hypertrophic cardiomyopathy and less frequently seen in dilated cardiomyopathy. A recent series of publications link variants in the 5’ end of the MYH7 gene as implicated in left ventricular noncompaction cardiomyopathy, often in the setting of a dilated ventricle with impaired function. Importantly, premature truncations as well as missense variation within the MYH7 gene has been linked to LVNC in both population studies and in individuals and families. We now generated a heterozygous premature truncation in MYH7 in human induced pluripotent stem cells (hiPSCs). When differentiated into engineered human heart tissues, we observe the heterozygous premature truncation in MYH7 produces a phenotype consistent with excess proliferation and reduced function, which are key features thought to underlie the development of LVNC in vivo. We hypothesize that truncations and missense variants identified in LVNC are associated with reduced contractility, rather than hyperdynamic MYH7 variants seen in hypertrophic cardiomyopathy. Additionally, many missense variants in MYH7 are considered variants of uncertain significance and methods such as those being used here may help adjudicate variants of risk. Through this training program under the K99 phase, Dr. Monroe will evaluate missense MYH7 variants associated with LVNC and evaluate their performance in engineered heart tissues. In his second aim, he will expand the search for LVNC-associated MYH7 variation to the population scale using linked cardiac imaging and genotype data in the in population datasets. As Dr. Monroe transitions to his independent phase, he will build from work performed earlier in his train implicating the Hippo pathway in proliferation and specification. In Aim 3, he will detail new disease relevance for the Yes-associated protein (YAP) in MYH7-associated LVNC using the models already in hand and further developed under his K99 training. Finally, in Aim 4, Dr. Monroe uses unbiased approaches to characterize human cardiomyocyte heterogeneity in healthy and LVNC engineered heart tissues in order to better delineate the range of differentiation and identify additional downstream pathways that will fuel future investigations. To promote his career development, Dr. Monroe will draw on the strengths of his mentoring committee and primary mentor which will focus on expanding his management and his own mentoring skills.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Intrahepatic cholangiocarcinoma (iCCA) is a devastating liver cancer with limited therapeutic options. While immune checkpoint inhibitors (ICIs) alone are ineffective in iCCA, the combination of ICIs with standard chemotherapy has recently become the new standard-of-care (soc). However, ~75% of iCCA patients present with primary resistance to the soc, emphasizing the critical need to identify underlying mechanisms of resistance and design more effective ICI combinations. Tumor genotype plays a critical role in iCCA immunogenicity, therapeutic resistance, and composition of the tumor microenvironment (TME), highlighting the need for precision models that (1) capture the genetic complexity of primary tumors, (2) reproduce patient-specific tumor- TME interactions, and (3) enable high-throughput immunotherapy screening. Current preclinical models fail to fully recapitulate these features. To address this gap, we have developed a novel co-culture system that allows the simultaneous reconstitution of iCCA patient-derived organoids (PDOs) with multiple autologous TME components, here called PDOTs. Our PDOT system integrates key stromal and immune components, providing a more physiologically relevant platform for immunotherapy testing. Our preliminary data show that iCCA PDOTs: (1) preserve the malignant programs and spatial cellular interactions of primary tumors; (2) reproduce real-life clinical response; and (3) identify surrogates of resistance to current ICIs that can be targeted to improve therapeutic efficacy. Based on these exciting preliminary data, our objective is to demonstrate the translational utility of our newly developed PDOT system in guiding the personalized design of effective immunotherapies that will ultimately improve the outcome, quality of life, and prognosis of patients afflicted by this devastating disease. Our central hypothesis is that PDOTs faithfully model the malignant programs and mechanisms of resistance to the soc chemoimmunotherapy in iCCA, and thus represent a valuable platform to test the preclinical efficacy of rational combination strategies able to improve patient responses. Thanks to the strategic position of Mt Sinai Hospital – a leading US center in the number of new liver cancer patients evaluated annually, including iCCA, – we are uniquely positioned to pursue the following Specific Aims. In Aim 1, we will establish a biobank of 50 iCCA PDOs and matched autologous immune and stromal components to assess their molecular and functional fidelity to primary tumors. In Aim 2, we will use PDOTs derived from 36 biopsies of iCCA patients prior to soc treatment to uncover cellular and molecular mechanisms of resistance and design rational combination therapies that improve patient responses. With over 70% of iCCA patients displaying primary resistance to the current ICI- based soc, investigating mechanisms of innate resistance presents a major unmet need in this disease. The completion of this study will provide an unprecedented framework to accelerate the development of precision immuno-oncology therapies for patients with iCCA and potentially other cancers, and open new horizons into translational studies aimed at developing more effective combination strategies in a patient-specific way.

NIH Research Projects · FY 2026 · 2026-05

Project Summary / Abstract Type 1 Diabetes (T1D) results from progressive destruction of pancreatic β-cells driven by autoimmune attack and intrinsic stress. While much attention has focused on immune-mediated mechanisms, the contribution of β- cell–intrinsic stress pathways to disease progression remains incompletely understood. Emerging evidence implicates ChREBPβ (Carbohydrate-Responsive Element-Binding Protein β), a stress-inducible transcription factor, as a central integrator of metabolic and inflammatory signals that impair β-cell identity and survival. Although ChREBP has been studied in the context of glucose metabolism and Type 2 Diabetes, its pathological role in T1D remains unexplored. Our preliminary data demonstrate that ChREBP and its target genes are upregulated in β-cells from autoantibody-positive (AAb+) and T1D donors, as well as in NOD mice, suggesting early activation in disease pathogenesis. ChREBPβ overexpression induces apoptosis, oxidative stress, and β-cell dedifferentiation. We also developed a novel small molecule, Compound 43, which functions as a “molecular glue” that stabilizes the ChREBPα–14-3-3 interaction, thereby suppressing ChREBPβ expression and protecting β-cells from stress- induced damage, and we demonstrate here that this stabilizer is able to protect human β-cell identity and function under cytokine-induced toxicity. This proposal aims to define the contribution of ChREBPβ to β-cell dysfunction in T1D and evaluate the therapeutic potential of Compound 43 in mitigating this process. In Aim 1, we will characterize the impact of ChREBPβ activation on β-cell stress and survival under inflammatory and ER stress conditions using human islets. We will assess gene expression, apoptosis, UPR activation, and lipid metabolism using transcriptomic and lipidomic profiling. In Aim 2, we will test whether Compound 43 can protect human islets and stem cell– derived β-cells from cytokine- and ER stress–induced dysfunction, using functional, metabolic, and transcriptomic readouts. This study will establish ChREBPβ as a previously unrecognized contributor to early β-cell failure in T1D and provide preclinical validation for a new pharmacologic approach that targets β-cell resilience, rather than immune modulation. These findings will offer a new conceptual and therapeutic framework for preserving β-cell function in the earliest stages of T1D, directly aligning with the mission of the Human Islet Research Network (HIRN) to understand and prevent β-cell failure.

NIH Research Projects · FY 2026 · 2026-05

Colorectal cancer (CRC) presents an immunological paradox: while CD8+ T cell infiltration is associated with better outcomes, most microsatellite stable (MSS) CRC cases remain resistant to immune checkpoint blockade (ICB), particularly in patients with liver metastases. This resistance suggests alternative immunosuppressive mechanisms, with myeloid cells—especially macrophages—playing a central role in suppressing anti-tumor immunity. Our research has identified two dominant macrophage programs, SPP1+ and IL-4-activated macrophages, that contribute to immune suppression and tumor progression in both human and murine CRC. Importantly, we found that cancer-associated stromal cells promote the accumulation of IL-4-producing eosinophils via IL-33, reprogramming macrophages into a repair-like phenotype that enhances tumor growth. We also found that GREM1+ cancer-associated fibroblasts (CAFs) drive the SPP1+ macrophage program, reinforcing a pro-tumorigenic feedback loop, potentially through IL-1 signaling. We hypothesize that myeloid- stromal crosstalks in the CRC tumor microenvironment (TME) drives immune suppression, stromal remodeling, and tumor invasiveness. To test this, we will: (1) define how the IL-4-myeloid axis promotes CRC progression; (2) dissect the pathogenic interaction between SPP1+ macrophages and GREM1+ CAFs; and (3) evaluate therapeutic strategies targeting these pathways to improve response to ICB. Using in vivo and ex vivo models, including orthotopic mouse tumors and human CRC tissue slices, we aim to establish the therapeutic potential of blocking IL-4Rα, IL-1R, IL-1RAP, and IL-33 pathways alone and in combination with ICB, to inform clinical trials for patients with oligometastatic CRC and liver metastases.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Perinatal or Postpartum Mood and Anxiety Disorders (PMADs) are leading causes of maternal morbidity and mortality, prompting great need to understand factors that influence risk. Parity, the state of having carried pregnancies to a viable gestational age, can promote positive parenting adaptations and/or increase the risk of developing PMADs, especially when compounded by chronic stress over the peripartum period. Yet, how stress and parity interact to result in long-lasting vulnerability is unknown. Therefore, the objective of this K01 proposal is to investigate the molecular and epigenetic mechanisms by which stress negatively interacts with parity programming during the critical postpartum period to influence future risk. Using a novel parity x postpartum stress mouse model, we show that parity alone leads to lasting positive adaptations in maternal and cognitive behaviors. However, the interaction of parity x postpartum stress results in increased anxiety-like phenotypes paired with loss of cognitive adaptations. Thus, we propose a framework in which parity promotes maternal brain plasticity to support behavioral adaptations, but that postpartum stress exploits this period of plasticity to induce lasting susceptibility. Using this model, we identify the dorsal hippocampus (dHpc) as a brain region exhibiting robust transcriptomic sensitivity to parity that may underlie maternal behavioral adaptations, both of which are disrupted by postpartum stress. Further, our data suggests that chronic postpartum stress dysregulates dopamine responsiveness to maternal stimuli via projections from the ventral tegmental area (VTA), underlying long-term perturbations of dHpc plasticity. Thus, under the mentorship of Dr. Ian Maze and the guidance of my Advisory Committee, I have designed three Specific Aims to address the hypothesis that epigenetic regulation of the VTA-dHpc projection alters dopamine signaling during postpartum, which in turn mediates the persistent impacts of parity and postpartum stress on maternal (mal)adaptations. In Aim 1, I will test the involvement of a putative parity-dependent transcription factor in regulating the VTA-dHpc response to postpartum stress by employing a combination of AAV viral vector strategies with transgenic mice, followed by functional and single cell epigenomic assessments. In Aim 2, I will determine the role of dHpc dopamine signaling in postpartum stress-induced acute and persistent behavioral alterations, and whether an additional pregnancy and postpartum exposure will induce maternal deficits and anxiety-like behavior during a subsequent postpartum window. In Aim 3, I will examine the involvement of the novel protein modification “dopaminylation” by using a novel chemical approach coupled with mass spectrometry to identify dopaminylated proteins that may mediate the long-term effects of parity and postpartum stress on dHpc plasticity. This research is critical for understanding how parity and stress interact to shape maternal brain health and PMAD risk, with implications for intervention strategies. Moreover, the training and career development activities in this K01 will provide the expertise and protected time I need to transition to an independent research career.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Pulmonary arterial hypertension (PAH) is a rare and progressive cardiopulmonary disorder characterized by the remodeling of pulmonary vascular cells, which leads to narrowing and the obstruction of pulmonary arteries. This results in increased pulmonary vascular resistance and pressure, ultimately causing right ventricular failure. Despite current therapies targeting key pathogenic pathways, the morbidity and mortality associated with PAH remain unacceptably high, highlighting the urgent need for new treatments. A hallmark pathological feature of PAH is the phenotypic shift of pulmonary artery smooth muscle cells (PASMCs) to a “cancer-like” state, which is characterized by hyperproliferation and resistance to apoptosis. Previous studies have suggested that abnormal histone methylation contributes to this pathological phenotypic switching, promoting uncontrolled vascular remodeling during the early stages of PAH. However, the precise molecular mechanisms involved are still poorly understood. Menin (Men1), a scaffold protein that interacts with the histone methyltransferase Mixed Lineage Leukemia 1 (MLL1 or KMT2A), plays a crucial role in regulating gene expression through epigenetic mechanisms, including the trimethylation of H3K4me3 histone marks, which influences oncogenic pathways and cell cycle control. While its role in cancer is well established, the involvement of the Men1/MLL1 complex in the pathogenesis of PAH has yet to be elucidated. In this proposal, we aim to elucidate the role of this complex by investigating how Men1 contributes to the phenotypic reprogramming of PASMC and vascular remodeling in PAH. Importantly, we will evaluate the therapeutic potential of Revumenib, a potent and selective Men1/MLL1 complex interaction inhibitor, in both in vitro and in vivo preclinical models of PAH. Our preliminary results show an upregulation of Men1 and MLL1 levels in both human PAH lung tissues and a rat model of PAH. Treatment with Revumenib led to a reduction in Men/MLL1 expression and interaction, accompanied by a decrease in H3K4me3 histone marks and reduction of PASMC hyperproliferation and migration. Based on these findings, we hypothesize that Men1 exacerbates vascular remodeling in PAH via MLL1-mediated H3K4me3 marks and associated transcriptional gene programs, thereby promoting the reprogramming of PASMCs. SA1 will evaluate the in vitro efficacy of Revumenib in PAH-PASMCs by examining its effects on cell proliferation, apoptosis, and gene expression signatures associated with Men1/MLL1 activity. SA2 will assess the in vivo therapeutic potential of Revumenib in healthy and in rat models of PAH. Biochemical and cellular assays will be used to evaluate drug distribution, Men1/MLL1 complex disruption, and downstream epigenetic modifications and signaling in the lungs. Given Revumenib’s promising safety profile in previous human and animal studies in acute myeloid leukemia, we anticipate minimal adverse effects in the context of PAH. The successful completion of this project will enhance our understanding of epigenetic regulation in PAH and may establish Revumenib as a foundation for the development of a novel, mechanism-based treatment strategy for this rare, deadly disease.

NIH Research Projects · FY 2026 · 2026-05

Prostate cancer remains the second-leading cause of cancer-related deaths in US men, mainly due to metastatic disease. Metastasis occurs most frequently in bones, thus entailing significant patient morbidity including pain, propensity to fractures and potential spinal cord compression. Moreover, the bone is a favored reservoir for undetectable disseminated tumor cells that maintain minimal residual disease and can thus critically define future patient outcomes. Despite this pressing clinical need, the mechanisms of progression to bone metastasis remain incompletely understood. Our overall goal is thus to understand the functional determinants of progression to lethal metastatic prostate cancer in order to develop more efficient therapies. Given that tumor progression and metastasis occur through multiple steps involving interactions with different benign cells and tissues, experimental models in which prostate cancer progression may be studied in a whole immunocompetent organism may help identify hitherto unappreciated mechanisms of progression. Our preliminary studies using novel mouse and human prostate cancer models show that ATAD2, an epigenetic and transcriptional regulator, is a critical mediator of metastasis (including bone) and of antitumoral immune responses. ATAD2 is progressively overexpressed during prostate cancer progression and may be an important therapeutic target because of its restricted expression in normal adult tissues as well as the presence of a potentially druggable and specific bromodomain. Furthermore, despite its widely reported association to worse survival in multiple cancer types, remarkably little is known about its functional role in metastasis. In this proposal we will determine the functional significance of ATAD2 expression for prostate cancer progression and metastasis. We will focus on its ability to modulate bone colonization and antitumoral immune responses, two critically relevant steps in the development of metastasis, and uncover the chromatin and transcriptional mechanisms through which it acts. Using state-of-the art syngeneic mouse models, ex-vivo epigenetic editing, human organoids and advanced tissue engineering technologies, our expert multidisciplinary team is uniquely poised to have a positive impact on our understanding of how tumor cells progress to lethal metastatic disease. Our studies will uncover novel mechanisms linking metastasis and immune escape, paving the way for future biomarker driven targeted therapies that may lead to durable and systemic therapeutic responses in currently incurable metastatic disease.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY How chronic type I interferon (IFN) signaling affects the lung epithelium and the connection between chronic interferon signaling, lung damage, and lung disease is poorly understood. IFN activation produces hundreds to thousands of interferon stimulated genes (ISGs), while the role of many ISGs is poorly understood. Dysregulated or persistent interferon responses, as seen in both type-I interferonopathies and severe respiratory infections, are linked to inflammatory lung pathologies, including interstitial lung disease and severe outcomes from viral infections. We aim to apply a novel proteomics technology, size exclusion chromatography coupled with mass spectrometry (SEC-MS), to the IFN system, adding a dimension of information to IFN system – protein interactions. This strategy seeks to identify new protein complexes and molecular "nodes" that modulate the IFN pathway response and may be disrupted in disease states. The proposed research aims to utilize: 1) SEC-MS and Affinity Purification Mass Spectrometry (AP-MS) to create a high resolution map of the protein interactions induced by IFN. This interactome will be functionally tested through targeted knockdown or overexpression of high-priority candidates – those that appear to amplify or suppress IFN signaling. In parallel, we aim 2) to use Human Airway Epithelium (HAE) cultures to model chronic IFN states, mirroring key features of interferonopathies and persistent interferon signaling in severe respiratory infections. Parallel study of the protein complexes formed in lung epithelial systems upon interferon activation and HAE cultures engineered to have persistent interferon signaling will deepen our understanding of how overt and dysregulated IFN signaling drives inflammation and lung damage. These studies could offer new insights for therapeutic interventions. The proposed research will be conducted under the joint mentorship of Dr. Jeffrey Johnson (Icahn School of Medicine at Mount Sinai) and Dr. Dusan Bogunovic (Columbia University). The dual expertise of both research groups will provide an excellent research environment to develop my skills as an independent scientist, comprising a wet lab and analytical and computational methodologies. The proposed training plan, which applies mentorship from Dr. Johnson and Dr. Bogunovic, guidance from my committee, and professional development from Mount Sinai, Columbia University, and the NYC area, will provide a strong professional and scientific launching point for my career as an independent scientist.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Cystic fibrosis (CF) is a multisystemic, autosomal recessive disorder with the majority of morbidity and mortality extending from lung disease. Given the benefits that older children and adults with CF have derived from cystic fibrosis transmembrane conductance regulator (CFTR) modulator therapies, it is anticipated that early childhood – or even in utero – treatment with CFTR modulator therapies may significantly delay or even prevent the development of CF lung disease. The role of CFTR protein in fetal lung development, and thus the potential impact of early CFTR modulator therapies, has yet to be fully elucidated. The proposed research project aims to better understand the pathogenesis of CF lung disease and the impacts of deficient CFTR protein on fetal lung development while overcoming a general roadblock in the study of many pediatric lung diseases, the scarcity of available human material. Previous immunohistochemistry studies in the fetal CF lung described a three-week delay in the developmentally regulated pattern of CFTR protein expression as well as an early, intrinsic, pro- inflammatory state. Leveraging a three-dimensional, in vitro and in vivo, lung organoid model that undergoes branching morphogenesis and alveologenesis – thus recapitulating early fetal lung development – we will characterize CFTR gene transcript expression and CFTR protein expression and function and articulate the transcriptome and proteome of normal and CF lung organoids. We hypothesize that CFTR protein deficiency in the fetal CF lung results in pro-inflammatory transcriptomic and proteomic profiles in respiratory epithelial cell populations and that this can be reasonably demonstrated using a lung organoid model. To test our hypotheses and validate the findings in the lung organoids, we will also articulate the cellular multi-omes in fetal normal and CF lungs, characterizing any disease-related cellular, transcriptomic, epigenomic, and proteomic changes. The proposed research project will provide critical insights into the pathogenesis of CF lung disease and establish a new, well characterized, in vitro and in vivo, model system of the CF lung that can be leveraged in future research endeavors (e.g. viral disease modeling, therapeutic testing). The proposed research project has the potential to identify the earliest pathogenic changes in the CF lung, determine the pulmonary impact of the application of in utero CFTR modulator therapies, and determine the need for alternative, novel treatment modalities to more readily change the early trajectory of CF lung disease.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Temporal lobe epilepsy (TLE) is a debilitating disorder that includes pervasive memory impairments that significantly impact quality of life, yet have no available treatment options. In both patients and rodent models of TLE, altered neural circuits lead to both hypersynchronous events like seizures, interictal epileptiform discharges (IEDs), and high frequency oscillations (HFOs), as well as desynchronization, with reduced coherence of oscillations and neural phase locking across regions. These changes in synchronization within and across brain regions are likely to disrupt normal cognitive function and contribute to memory impairment. One important mechanism of synchronization in the healthy brain is the dentate spike, a large-amplitude event that can occur along with synchronous neural activity across the brain. Yet, little is known about how these events are initiated, how they drive synchronous neural activity, and how they contribute to spatial memory. In Aim 1 of this proposal, we will use in vivo electrophysiology with silicon probes to characterize how neural activity is synchronized across medial entorhinal cortex (MEC) and hippocampus during type 2 dentate spikes (DS2s), which are hypothesized to be driven by inputs from layer 2 stellate cells in MEC. We will optogenetically identify these MEC2 stellate cells and both stimulate and inhibit them to determine their causal role in DS2 generation and neural synchronization. Then, in Aim 2, we will examine how DS2 rates and neural synchronization are altered in the pilocarpine-induced status epilepticus mouse model of TLE. In addition, we will record and directly manipulate MEC2 stellate cells during a cognitive task to determine their causal role in spatial cognition, and how they mediate memory impairment in epilepsy. Together, these Aims will determine the precise neural circuits that drive DS2 events and associated neural synchronization, and test how these events and synchronization processes are disrupted in chronically epileptic mice.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY Preliminary Data, based on siRNA knockdown studies, suggests that host encoded DExD/H box family protein DDX60L exerts anti-Ebola virus (EBOV) activity. We therefore hypothesize that DDX60L is a negative regulator of EBOV replication that can influence EBOV disease (EVD). While constitutively expressed in a number of cell types, DXX60L is an interferon (IFN) stimulated gene (ISG) and is related to DDX60, which is reported to augment production of IFNα/β. Therefore, DDX60L may also contribute to the anti-EBOV effects of IFNs and promote upregulation of IFN responses. As a negative regulator of infection, understanding the mechanisms of DDX60L-mediated inhibition may suggest strategies to mitigate EVD. We proposed to define how DDX60L exerts its anti-EBOV effects using a combination of siRNA knockdown and CRISPR-Cas9-based approaches. Studies will be performed in cell lines that correspond to cell types infected by EBOV in vivo. These are the Huh7 (hepatocyte), A549 (epithelial) and THP-1 (monocyte/macrophage) cell lines. Because siRNA knockdown measurably impairs EBOV growth in cell culture, we will use this approach to define effects on a transfection- based EBOV replication cycle modeling system that can assay all the major steps in the virus replication cycle. In parallel, we will use in knockdown studies a recombinant EBOV in which the viral VP30 coding sequences have been replaced with GFP (EBOV-GFPΔVP30). This virus only replicates in cells that provide VP30 in trans, and can be used at reduced biocontainment levels. Finally, we will assay the effects of knockdown in the context of fully replication competent EBOV. For each system, we will perform assays in the absence and presence of IFNα and define steps in the replication cycle affected by knockdown using a combination of approaches that include measuring viral RNA synthesis, viral protein production and host IFNα/β responses. We will also build additional experimental systems to further characterize the role of DDX60L in EBOV infection. First, we will generate knockout cells using CRISPR-Cas9. This will allow us to define the impact of DDX60L more definitively. Second, because DDX60L is an enzyme, we will use CRISPR base editing to mutate the ATP binding site in the endogenous gene, to determine whether enzymatic activity is required for anti-EBOV effects. Finally, we will use a CRISPR-Cas9 activation approach to induce expression of endogenous DDX60L and determine whether this enhances its antiviral effects. This latter approach will enable over-expression studies in difficult to transfect cell types and overcome reported difficulties of expressing DDX60L from mammalian expression plasmids. Multiple alternatively spliced DDX60L mRNAs have been described. By inducing expression of DDX60L from its endogenous promoter, the CRISPR activation approach is expected to yield normal splicing patterns, overcoming the need to select a specific isoform for study. As these new cell-based systems become available, they will be used in the assays developed to study DDX60L-EBOV interactions in the context of siRNA knockdown.