Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Psychiatric disorders remain a leading cause of disability in the US and are associated with increased morbidity and mortality. Early detection and treatment is essential to improving long-term outcomes, yet a substantial proportion of patients with psychiatric complaints experience long diagnostic odysseys before receiving an appropriate diagnosis and initiating effective treatment. “Learning health care systems” aim to short-circuit this slow process by leveraging the diagnostic, treatment, and utilization patterns left behind in “big data” (e.g., clinical, genomic, and social determinants of health) to more efficiently and accurately match the right patient with the right diagnosis/treatment, at the right time. Furthermore, over the past several years, a new paradigm– precision medicine–has moved to the forefront of biomedical research and clinical practice. Precision medicine has been defined as “an approach to disease treatment and prevention that seeks to maximize effectiveness by taking into account individual variability in genes, environment, and lifestyle.” Since its inception in 2018, the mission of the PsycheMERGE network has been to advance precision psychiatry in a learning health care system framework. This application, which was developed collaboratively by PsycheMERGE Network members, represents an opportunity for profound advancement of both basic and translational research in precision psychiatry. We propose extending our foundational efforts to now address barriers to scalability, utility of genomic data, clinical application, and translation to clinical practice in a precision psychiatry paradigm. Specifically, Aim 1 creates a nation-wide federated transfer-learning platform for the development of generalizable and bias-aware algorithms. Aim 2 integrates state-of-the-art methods to perform inclusive trans-ancestry genomic analysis of biobank samples and further innovates by leveraging the breadth and depth of medical record data to discover novel biology that can further inform precision psychiatry paradigms. Aim 3 addresses the application of algorithms by focusing on two use cases including (a) differential diagnosis between bipolar disorder 1 and other mood disorders, as well as (b) probabilistic treatment response to antidepressants for acute depressive episodes. Lastly, Aim 4 uses mixed methods to assess the feasibility, utility, and attitudes towards precision psychiatry tools. Our combined sample of clinical EHR data exceeds 29 million individuals and of those, nearly 2 million also have genetic data already available for analysis across the twelve sites included in this application. A cross-cutting theme throughout the application is the intentional focus on equitable performance of algorithms, innovative integration of social determinants of health, and inclusive methods for genomic analyses. The sites included are also representative of many diverse communities across the United States including the East and West Coasts, the South, and the Midwest. This application represents a major step towards equitable precision psychiatry and brings the field closer to the goals outlined in the updated NIMH Strategic plan.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY/ABSTRACT The research proposed in this application aims to study the impact of the publication of the 2016 Centers for Disease Control and Prevention (CDC) analgesic prescribing guidelines on disparities in opioid utilization among older adults who have opioid-responsive pain. These CDC guidelines were published at that time to help curb the rising number of overdose-related deaths related to prescription and illicit opioids, which have caused the death of over half a million people in the United States since 1999. The guidelines recommended cautious use or avoidance of opioid medications in persons with chronic, non-cancer pain and those not near the end of life. However, preliminary evidence suggests that these guidelines may have had an unintended effect of limiting access to legitimate and medically necessary opioids in people with serious illnesses. This is of particular concern among vulnerable subgroups of older adults known to be risk of pain undertreatment. Our analyses will focus on three highly vulnerable subgroups of older adults who have been found to be at increased risk of pain undertreatment – persons with Alzheimer’s and related dementias (PW-ADRD), those identifying as racial/ethnic minoritized individuals, and those experiencing socioeconomic deprivation. For each of these three subgroups, we will employ two models of opioid-responsive pain conditions – chronic cancer pain and acute pain from orthopedic trauma – for which the CDC guidelines were not intended to apply and for which opioids remain a mainstay of therapy for pain control. These two conditions are common in older adults and are often associated with moderate to severe pain. For each of the three subgroups of older adults, we will utilize the Medicare Current Beneficiary Survey (MCBS) to obtain sociodemographic and clinical information on opioid prescribing from 2010 to 2020. MCBS is a longitudinal survey of a nationally representative sample of the Medicare population, with data collected three times per year over a 4-year period and approximately 15,000 respondents each year, and includes linked fee-for-service Medicare Part A and B claims, participant- reported prescribed medicines, and records of Medicare Part D pharmacy events. We will use a difference-in- differences design to compare the unintended impact of the 2016 CDC guidelines on opioid utilization from 2010-2020 in the following groups of older adults who have either cancer or hip or pelvic fracture: PW-ADRD compared to persons without ADRD (Aims 1-3), those with self-reported income at or below the federal poverty line, compared to individuals with higher income (Aim 2), and racial/ethnic minoritized older adults compared to white older adults (Aim 3).

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT The present crisis of opioid addiction in the United States highlights how an initial exposure to opioids, often oxycodone, as a medical patient can lead to escalation of use, misuse and eventually addiction. Chronic exposure to oxycodone produces tolerance to many of its effects, and it is this development of tolerance that leads to taking increasingly higher doses and to withdrawal upon abstinence. Drug-induced adaptations in gene expression and neuronal function are thought to underlie the development of tolerance and withdrawal, but the mechanisms by which this occurs remain unclear. We have found that the habenula-interpeduncular nucleus (IPN), which plays a prominent role in limiting nicotine intake and in nicotine withdrawal responds to chronic nicotine by upregulating nitric oxide synthase 1 (NOS1), and that this same adaptation occurs in response to chronic oxycodone exposure. Acute exposure to oxycodone is not sufficient to upregulate NOS1 in the IPN. Using viral approaches, purification of tagged ribosomes followed by next gen sequencing and oral oxycodone in the drinking water of adult male and female mice, we will examine gene expression in the IPN – both in the presynaptic terminals and postsynaptic IPN neurons - and correlate neuronal calcium dynamics with intoxication and withdrawal behavior. We will also examine the contribution of NOS1 to drug-induced changes in gene expression in neuronal activity and axon terminals from the medial habenula within the interpeduncular nucleus. Together these aims will establish a preclinical model of opioid tolerance that may provide a basis for evaluating potential new therapeutics in the treatment of opioid use disorder.

NIH Research Projects · FY 2025 · 2024-09

Alzheimer's disease (AD) remains uncurable. The complex, and diverse, neuropathology suggests that AD may actually represent not a single disease, but a family of diseases that share plaque and tangle neuropathology. The overall modest effects and variability among patients' response to the recent FDA approved drugs for the treatment of AD highlight potential disease heterogeneity among study subjects. The high heterogeneity of AD has also been strongly supported by the recent discovery of three major molecular subtypes of AD, each possessing distinctive molecular signatures. The therapeutic strategy, referred to as “senolytics,” has gained immense research attention for its hope to improve various age-associated conditions, including AD and related dementias (ADRD), by pharmacologically removing senescent cells in the brain. Based on our preliminary findings, there is compelling evidence indicating that senescence exhibits distinct characteristics in the major molecular subtypes of AD. In this application, we propose to systematically investigate the molecular mechanisms of brain senescence in different molecular subtypes of Alzheimer's Disease (AD) for discovery of novel targets and therapeutics for AD. The project will be carried out by a multidisciplinary team of leading scientists with expertise across translational Sen biology, neuropathology, bioinformatics, systems biology, machine learning and artificial intelligence. We have successfully piloted, and here will deploy, a Sen multi-Omics (senomic) pipeline to define a scale molecular the holistic cellular, molecular and chemical phenotypes of Sen cells in brains from large number of AD and control subjects. In this project, we will systematically analyze all the existing large- single cell RNA-seq and multiome data in AD to identify and characterize senescent cells in major subtypes of AD. We willvalidate key findings using single-cell multi-Omics and spatial transcriptomics and imaging in which the entire human transcriptome can be profiled in single cells while maintaining spatial and multi-scale resolution. Using our unique pipeline, we are very well positioned to characterize and quantify the molecular heterogeneity of Sen cells in different AD subtypes. Our iterative approach involves profiling intact tissues by 10xGenomics Visium and NanoString GeoMx and CosMx platforms as well as disaggregated cells by single nucleus multi-Omics (RNA-seq and ATAC-seq). Results will provide a spatially resolved, comprehensive molecular portrait of both chromatin accessibility, gene and protein expression in the hippocampus in AD in contrast with healthy control. Our methodologies are non-destructive allowing for mapping multi-analytes back to the tissues to determine cellular morphology and neighborhood environment. We will further develop novel therapeutics against brain senescence in AD through cutting edge drug repositioning approaches. Key driver genes and candidate drugs will be validated through extensive in vivo experiments. The data and the analytic and experimental tools will be shared with the community for broader investigations.

NIH Research Projects · FY 2024 · 2024-09

Visceral pain is a key feature of functional gastrointestinal (GI) disorders such as irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), and obstructive bowel disorders (OBD). Notably, there is also a high comorbidity of chronic visceral pain with stress-related psychiatric disorders including anxiety, depression, and fatigue, significantly impairing patients' quality of life. However, the molecular and circuit mechanisms of visceral pain are poorly understood, and the resulting lack of effective clinical management represents a major unmet medical problem. Pilot studies showed that activation of the glutamatergic Tacr3+ neurons in the spinal cord dorsal horn (SDH) is sufficient to drive both sensory discriminative (visceromotor response) and affective (aversion) visceral pain responses. Additionally, the parabrachial nucleus of the pons (PBN) in the brainstem had the most abundant projections of the Tacr3+ SDH neurons. These observations led us to frame the hypothesis that the glutamatergic Tacr3+ SDH neurons are excitatory interneurons and projection neurons mediating visceral pain processing from the mouse colon to the brainstem. The objective of this R56 is to elucidate whether the neural circuits from the colon-innervating spinal visceral afferents form functional synaptic connections with the Tacr3+ SDH neurons and dissect the function of Tacr3+ spinal projections in the brainstem. We will demonstrate for the first time that the glutamatergic Tacr3+ neurons receive synaptic inputs from the colon-innervating spinal visceral afferents, and stimulation of the colon-innervating spinal afferents is sufficient to activate the Tacr3+ SDH neurons. We will also investigate whether activating the Tacr3+ spinal projections in the PBN drives aversive behavior. The outcomes of these studies will advance our understanding of the cellular and circuit mechanisms of visceral sensory processing in the lumbosacral

NIH Research Projects · FY 2025 · 2024-09

SUMMARY Transcription factors play essential roles in establishing the correct gene expression patterns unique to each cell type. These finely controlled patterns can fall apart if a mutation resides within a critical factor. In the hematopoietic system, this can lead to anemia, dysplasia, or leukemia of varying morbidities. Overcoming transcription factor deficits is highly challenging, especially if the mutation is solely in one of the two alleles and if complete removal of the protein’s expression is not an option. Experiments in this proposal addresses the problem in the context of the congenital dyserythropoietic anemia (CDA) caused by a single missense mutation in one allele of the KLF1(EKLF) transcription factor. Technological development of state-of-the-art approaches to selectively target and degrade the mutant variant will be designed and tested. Specifically, the method relies on the cellular introduction of degradative modules (degrons) coupled to targeting motifs based on KLF1/DNA interactions, the details of which are to be based on published and proposed methods of identifying parameters of binding selectivity. These studies, summarized in two Aims that cover (1) in vitro tests and (2) in vivo (cell culture) approaches, will have a transformative effect well beyond the immediate case, as they provide an innovative and distinctive blueprint for the direct removal of problematic proteins in other hematologic and renal diseases caused by monoallelic missense mutations in transcription factor DNA binding domains.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Pulmonary arterial hypertension (PAH), a deadly cardiopulmonary disease that develops following a genetic or environmental insult, is characterized by pulmonary endothelial cell dysfunction and occlusive vascular remodeling. The loss or mutation of the Bone Morphogenetic Protein Receptor Type 2 (BMPR2) function, which regulates the growth and differentiation of pulmonary vascular cells, plays an important role in PAH. Switch-Independent 3a (SIN3a) is a master transcriptional scaffold that plays an essential role in regulating gene transcription and maintaining chromatin structure, and its absence or inappropriate recruitment has been associated with aberrant gene silencing. We have recently shown that SIN3a plays a central role in the hypermethylation of the BMPR2 promoter in pulmonary artery smooth muscle cells and the pathogenesis of PAH. Interestingly, BMPR2 is highly expressed in pulmonary artery endothelial cells (PAECs) and is vital to their function and survival. The molecular mechanisms of SIN3a epigenetic regulation underlying BMPR2 gene alteration in PAH-PAEC are not fully elucidated, thus, identifying the mechanism(s) that restore SIN3a expression/function in PAECs in the setting of PAH will offer new therapeutic opportunities. Our objectives in this proposal are to elucidate the role and the downstream mechanisms of SIN3a on the regulation of BMPR2 and EC dysfunction in PAH and to evaluate the therapeutic effects of targeting PAECs using lipoprotein nanoparticles encapsulating modified RNA encoding SIN3a (SIN3a.modRNA-LNP) and AAV6.hSIN3a gene therapy. While investigating how SIN3a regulates BMPR2 in PAECs, we discovered a novel molecular pathway by which SIN3a overexpression inhibits the Enhancer of Zeste Homolog 2 (EZH2) and reverses FOXK2 promoter region histone methylation and restores FOXK2 and consequently upregulates BMPR2 by increasing FOXK2 binding to the BMPR2 promoter. Based upon these findings, we hypothesize that SIN3a plays a critical role in PAEC homeostasis; mechanistically, disrupting SIN3a expression contributes to FOXK2 promoter histone modifications and thus to BMPR2 dysregulation and EC dysfunction, resulting in PAH. In this proposal, the hypothesis will be tested by pursuing the following specific aims: Aim1) To define the role of SIN3a and the epigenetic mechanisms in regulating the BMPR2 gene and PAH-PAEC dysfunction. Aim 2) To characterize SIN3a/FOXK2 interplay in BMPR2 regulation in the setting of PAH in human lung tissues and SIN3a KO mice. Aim 3) To evaluate the therapeutic efficacy of targeted SIN3a gene transfer to lung endothelial cells in small animal models of PAH. The proposed study presents the novel concept that modulation of SIN3a is essential for endothelium function in PAH disease and applies a systematic approach to integrate sophisticated functional validation assays, modified mRNA, and AAV6 gene therapy. Significantly, the proposed research is expected to vertically advance and expand our understanding of epigenetic regulation in pulmonary endothelial cells in PAH. Such knowledge is critical for the development of PAH therapies targeting epigenetic aberrations.

NIH Research Projects · FY 2024 · 2024-09

Project Summary The promise of genomic medicine to transform healthcare and improve health will not be fully realized until discoveries become relevant to and available for use by diverse populations and their clinicians. As part of the IGNITE II network, we are implementing two prospective randomized pragmatic genotype-guided clinical trials (GUARDD-US and ADOPT-PGx) to determine the impact of implementing genetic testing on hypertension, depression, and pain therapies. This administrative supplement request is to extend the implementation timeline for these two pragmatic clinical trials by an additional 12 months with cost to allow for completion of GUARDD-US and ADOPT PGx data analysis, writing up of study results and closeout of all study activities. We expect the successful results from this clinical trial will provide critical evidence needed to drive the implementation of genomic medicine across broad demographics of patient populations.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Autism Spectrum Disorder (ASD) risk is primarily of genetic origins with an estimated heritability of 80% but non-heritable factors are also important in understanding the underlying etiology. One source through which environmental effects may be influential is preterm birth (birth before 37 weeks gestation) which constitutes 10% of all births in the US. The etiology of preterm birth is primarily of environmental origin, usually due to disease in the pregnant woman or her fetus. The processes underlying preterm birth may initiate the development of ASD. This hypothesis has not been previously studied, and if true the heritability of ASD may be lower in children born preterm, and before the optimal gestational age for delivery at week 39 to 40, suggesting that children born preterm define a subset of the general population where environmental risk plays a larger role for ASD risk. The goal of this project is to examine the etiology of ASD in children born preterm and to determine if the etiology is different compared to term-born children, and across gestational age; to identify risk factors for ASD in PTB children, compared to term born, and to estimate the public health consequences of these risk factors, how much can be attributed to confounding factors instead of the risk factors per se and if there are differences in genetic architecture. To achieve this goal, we will create a prospective cohort of all children born in Sweden and Finland 1996 to 2020 followed for clinical diagnosis of ASD, and link information from Swedish and Finnish National registers until the end of 2024. We will replicate analyses in a sample from California and examine genetic markers in two large genetic databases for ASD research. In Aim 1 we will determine the epidemiology of ASD in children born preterm, and characterize the risk factors for these children. Aim 2 will estimate population measures of risk (heritability and proportion susceptible) and estimate public health consequences of the risk factors from aim 1 by calculating the proportion attributable fractions for the risk factors. Aim 3 will replicate the analyses in aim 1&2 in a similar database from California. Aim 4 will test if the genetic architecture of ASD differs in preterm and term-born children.

NIH Research Projects · FY 2025 · 2024-09

Summary/Abstract Parkinson’s Disease (PD) is the second most common neurodegenerative disease affecting nearly one million people in the United States, and up to 80% of PD patients eventually develop Parkinson’s Disease Dementia (PDD), many with co-Alzheimer’s Disease pathology. PD is a neurodegenerative disease impacting the dopaminergic cells in the substantia nigra pars compacta (SNpc), which play a large role in controlling both movement and cognition. The motor circuit involves the lateral SNpc projections to the striatum, while dementia phenotypes involve the medial SNpc projections to various cortical and limbic related structures. To date, there are no significant therapeutics that prevent progression of disease and dementia, highlighting a critical need for further understanding of PD pathology. Notably, epigenetics has gained prominence, with histone deacetylase inhibitors showing promise for neurodegenerative diseases, including PD. Recently, H3K4me3, trimethylated histone 3 at lysine 4, has come to light as a neuroprotector in a drug induced rat model of PD. This proposal aims to interrogate a novel epigenetic marker, dopaminylation, in PD. Our lab recently established a novel role for dopamine termed dopaminylation, whereby dopamine acts as an epigenetic marker on Histone 3 glutamine 5 (H3Q5dop), adjacent to the significant H3K4me3 modification. H3Q5dop is covalently attached via the Tissue- Transglutaminase 2 enzyme to proteins and this process is concentration dependent. Given the loss of dopamine in PD, there is indeed an observed loss of H3Q5dop in postmortem human PD SNpc tissue and a global loss of dopaminylation observed in postmortem human PD post-synaptic BA25 cortical tissue. Interestingly, the loss of H3Q5dop was found in conjunction with and independent of the neighboring H3K4me3 epigenetic modification. Preliminary findings suggest that H3Q5dop plays an important role in preserving and potentiating H3K4me3 readout, possibly pointing to H3Q5dop’s role in neuroprotection as well. Thus, I hypothesize that H3Q5dop influences chromatin architecture via enhancing H3K4me3 signal, and its decrease in PD alters cellular identity, SNpc projections to key motor and cognitive brain regions, and behavior. Firstly, this proposal will interrogate H3Q5dop in human postmortem SNpc and BA25 prefrontal cortex via the integration of data from epigenetic regulation, chromatin accessibility, and gene expression. Secondly, in mouse models, this proposal will assess the loss of H3Q5dop mediated changes to transcriptional identity, SNpc projections, motor function, and emotion-based learning. This innovative work will offer novel insights into H3Q5dop’s role in the pathophysiology of PD, establishing a novel therapeutic target. This proposed research will take place at the Icahn School of Medicine at Mount Sinai, within the Friedman Brain Institute, one of the world’s premier institutions for translational brain and nervous system sciences. With hopes of becoming a leading physician neuroscientist, working towards the proposed training goals will equip me with essential skills in molecular, behavioral, and computational research while advancing my medical training.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Despite advances in our understanding of rare genetic diseases and their causes, only 8% of these diseases have targeted drugs. Much of this arises from the disconnect between the inhibitory nature of drug molecules and a predominance of loss-of-function mechanisms in such diseases. There has been a growing appreciation of the role of gain-of-function variants in this context, especially towards drug repurposing. More specifically, we and others have shown that even subtle changes in function such as alterations of post-translational modification or molecular interaction sites can frequently lead to such disorders. It is unclear to what extent these observations generalize and are actionable from a therapeutic perspective. Common Fund data sets such as those from the Gabriella Miller Kids First, Undiagnosed Disease Network, the Illuminating the Druggable Genome and LINCS programs provide a unique opportunity to assess this computationally. Our central hypothesis is that gain-of-function variants account for a much larger proportion of rare genetic diseases than currently known and in silico functional profiling can be used to computationally identify such diseases. The proposed work will test this hypothesis through two aims. In Aim 1, we will apply our previously- developed predictors of variant impact towards the identification of known and predicted disease-associated variants in large Common Fund genomic data sets. In Aim 2, we will subset out those variants that impact druggable biochemical properties either directly or indirectly, to thus, infer novel drug-disease pairs. Over the award period, the principal investigator (PI) will leverage his and his team's expertise in variant interpretation, machine learning and bioinformatics knowledgebases towards the systematic integration of genomic and drug- related data from multiple Common Fund data sets to identify candidate drugs that can be repurposed for rare genetic diseases. This work will be carried out at the Icahn School of Medicine at Mount Sinai, home to world- renowned researchers in human disease genetics, robust computational infrastructure, and a thriving biomedical data science training environment. The proposed research will not only provide valuable pilot data for experimental validation of promising drug repurposing candidates but will serve as the foundation for future computational methodology development that will expand the scope of variants and mechanisms that can be queried. The work is expected to have broad impact, as it presents a new mechanism-centric, data-driven approach to identifying drug repurposing candidates for rare genetic diseases, that is generalizable to other situations.

NSF Awards · FY 2024 · 2024-09

The broader impact of this I-Corps project is based on the development of an artificial intelligence-driven clinical decision support system to detect endometriosis. Endometriosis is a chronic medical condition in which tissue similar to the lining inside the uterus, called the endometrium, begins to grow outside the uterus. This condition can cause pain, irregular bleeding, and may lead to infertility. Early diagnosis of endometriosis can improve patient outcomes, increase access to care, and lead to more treatment options. This technology combines patient screening with image analysis to provide a comprehensive clinical decision support system for physicians and radiologists caring for patients at risk of this condition. This innovative approach to early-stage endometriosis detection has the potential to reduce diagnostic disparities and democratize access to high-quality care for all women. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of the technology. The solution is based on the development of a clinical decision support system that offers a non-invasive, objective, and accessible solution for early-stage detection of the superficial peritoneal lesions found in endometriosis. The technology combines a patient screening tool and an imaging analysis tool to identify key health indicators and risk factors to help primary care physicians and gynecologists detect early-stage endometriosis. The technology also analyzes magnetic resonance imaging (MRI) images based on a deep learning model with more than 5 years of high-quality patient data, offering radiologists a more transparent, reliable, and repeatable scoring mechanism for potential endometriosis. 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 2024 · 2024-09

Project Summary/Abstract The serotonergic (5HTergic) system is implicated in a wide range of neurodevelopmental and neuropsychiatric phenomena, including regulation of mood and stress reactivity. While 5HT actions have been assumed to be mediated exclusively through 5HT receptors, studies have established 5HT forms covalent bonds with histone H3—resulting in H3 glutamine 5 serotonin (H3Q5ser)–a process known as H3 serotonylation. H3 serotonylation plays key regulatory roles in establishing normal embryonic and adult patterns of brain transcriptional plasticity. However, functional roles for H3 serotonylation during postnatal brain development, have been unexplored, and the impact of environmental stimuli on this modification during early life remains unknown. Our laboratory implemented FANS coupled CUT&RUN (C&R)-seq to profile the cell type-specific epigenomic landscape of H3 serotonylation in mouse medial prefrontal cortex (mPFC), across critical periods of postnatal brain development. We identify cell-type-, sex-, developmental-, and early life stress- (ELS) induced alterations in H3 serotonylation genomic enrichment, with males showing more pronounced developmental and ELS-induced changes in both neuron and glia populations. In males, further analysis reveals, H3 serotonylation binding increases at oligodendrocyte genes in response to ELS. While these findings are insightful, they alone cannot elucidate the cell-type-specific functional roles of H3 serotonylation on gene expression or its potential influence on behavioral phenotypes. In the F99 phase, I will test the hypothesis that H3 serotonylation critically regulates neurodevelopmental gene expression, and perturbation by ELS results in altered gene expression and increased vulnerability to stress-related behavioral phenotypes as a function of this novel PTM. I will utilize multi-omic bioinformatic tools to integrate epigenomic and transcriptomic (ie. RNA and C&R-Seq) data to uncover the causal link between H3 serotonylation and gene expression during critical postnatal brain development and in response to ELS. Using a novel cell-type specific viral construct, I will attenuate ELS-induced H3 serotonylation changes, to elucidate the functional cell-type specific roles of H3 serotonylation in brain development and its connection to stress-related behaviors in adulthood. During this phase, I will receive technical training on large-scale data analysis, integration of cell-type specific “omic” data, and molecular cloning techniques that can be used to create gene editing tools to manipulate epigenetic modifications such as H3 serotonylation in vivo. In the K00 phase, I will focus on identifying laboratories for my postdoctoral work, prioritizing expertise in circuit-specific approaches (circuit characterization/manipulation), functional readouts of neuronal physiology (fiber photometry), and use of computer vision and machine learning to unbiasedly quantify animal behaviors. Together, this training across F99 and K00 phases will support my successful transition to postdoctoral researcher and ultimately a career as an independent research scientist focusing on how circuit level changes following stress exposure intersect with epigenomic and transcriptomic changes in development and how these changes play in psychiatric disease.

NIH Research Projects · FY 2025 · 2024-09

Abstract The gametes arise either from germ cells or germline stem cells (GSCs), by undergoing differentiation and meiosis. The gametes help launch the next generation by sustaining early development prior to zygotic genome activation by contributing specific components to the zygote. In particular, the mature oocyte, in addition to a haploid genome, contains a “maternal contribution” of RNAs, proteins, complexes, and organelles that is critical to establish a totipotent zygote. Moreover, the oocyte cytoplasm is also sufficient to reprogram a quiescent somatic cell nucleus to totipotency. Thus, the oocyte's maternal contribution is essential and critical for totipotency. Germ cells undergoing differentiation and meiosis to eventually synthesize maternal contribution is conserved up to human. Nevertheless, the mechanisms that establish this highly potent oocyte cytoplasm during oogenesis are incompletely understood. Using Drosophila as a model system, my lab has discovered a germ cell-to-maternal transition, wherein germ cell-specific programs, such as those that promote GSC self-renewal, differentiation, and entry into meiotic cell cycle, are silenced once the oocyte is specified. Our work reveals that this silencing of the germ cell program is critical to establishing the “correct” maternal contribution by excluding transcripts that could interfere with development of the early embryo. We find that this silencing is is mediated by heterochromatin formation to and removal of perduring RNAs by RNA degradation pathways. Although, we are beginning to understand how germ cell specific programs are silenced during the germ cell-to-maternal transition several aspects remain poorly characterized. The objective of the proposal is to uncover how the germ cell-to-maternal transition is regulated by addressing the following questions: 1. How do the changes in gene expression during the germ cell-to-maternal transition relate to reorganization of the genome?, 2. How are germ cell-specific genes targeted for silencing? and 3. How is silencing of germ cell-specific genes coordinated with oocyte specification? The rationale for the proposed work identifying regulators of this transition and the underlying molecular mechanisms could reveal new etiologies and therapies for human infertility and novel concepts in developmental biology.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The overarching goal of this K08 research project is for Dr. Ethan Abbott, principal investigator (PI), to establish himself as an independent physician-scientist whose research addresses healthcare-related disparities and improves survival for out-of-hospital cardiac arrest (OHCA) patients. He has prepared with assistance of his multi-disciplinary mentorship team a compelling and innovative research project with matching career development training components that enables him to conduct this research, prepare and submit his subsequent R01-supported project from the preliminary data, and launch his research career. Dr. Abbott’s K08 research project aims to improve OHCA survival and clinical outcomes by identifying important individual-level health-related social needs (HRSN) domains to improve prediction of 30-day survival after OHCA and survival to hospital discharge. Despite the importance of individual-level HRSN in health outcomes, current OHCA predictive models only account for clinical variables, resulting in significant limitations towards advancing health equity and improving care for patients. Use of data science techniques, such as natural language processing (NLP) and large language models (LLMs) to identify and extract HRSN for inclusion in predictive models, could lead to interventions that decrease OHCA mortality. The specific aims of Dr. Abbott’s K08 research project are to: (1) Create a baseline predictive model to identify key pre-hospital, patient-level, hospital-level and clinical predictors of 30-day survival after OHCA and survival to hospital discharge; (2) Evaluate the efficacy of NLP and LLMs to extract individual-level HRSN for the OHCA cohort; and (3) Determine if inclusion of individual-level HRSN increase performance of the baseline predictive model in predicting 30-day post-OHCA survival and survival to hospital discharge, and if the HRSN- inclusive model performs better than the prior models NULL-PLEASE and CAST. He will rigorously develop the models using mediation analyses, multivariable regression, and machine learning algorithms. Dr. Abbott’s career development plan builds on his experience as an emergency medicine physician and junior faculty researcher to develop and acquire new skills and expertise in: (1) formal mediation analysis; (2) application of NLP algorithms and LLMs for electronic health record information (EHR) extraction, particularly for HRSN; (3) predictive analytics for clinical outcomes using machine learning algorithms; and (4) research independence through professional development activities, including committee leadership positions, grantsmanship, dissemination of research findings. The results of this K08 will generate preliminary data to form the basis of Dr. Abbott’s subsequent R01 application submission. His R01 study will externally validate a predictive model for OHCA and extraction using LLMs for individual-level HRSN. The public health importance of this work is that it will contribute overall to improved OHCA clinical care and survival for patients and evaluating best practice for inclusion of HRSN data in clinical care.

NIH Research Projects · FY 2025 · 2024-09

Functional and molecular characterization of Oncofetal Stem Cells in CRC Project Summary. Background. Colorectal cancer (CRC) is the second most deadly cancer worldwide, with nearly one million deaths annually1. CRC develops as a result of accumulating genetic and epigenetic aberrations in the colonic epithelium leading to formation of benign adenomas that evolve to invasive adenocarcinomas 2,3. Despite the advances in treatment options, resistance to therapy and recurrence remain inevitable in patients with advanced disease, accounting for the high mortality rates 4,5. The underlying malignant features have been often attributed to the LGR5+ CSCs. However, the unexpected findings that certain LGR5- cells sustain tumor growth upon selective ablation of this population 6,7 are consistent with the recent description of a potential second stem cell population, with fetal-like characteristics 8-10. While this is intriguing from a therapeutic perspective, existence of a second, plastic, stem cell population in a more primitive state (hereafter referred to as OnFSCs for OncoFetal Stem Cells) is solely based on transcriptional signatures. Gap. Despite recent studies describing its existence, the molecular underpinnings, functional relevance, and therapeutic implications of the OnF cellular state in CRC remain largely unexplored. Hypothesis. We hypothesize that OnFSCs work in tandem with the LGR5+ stem cells (SCs) to fuel tumor evolution in CRC. OnF cells emerge early during intestinal tumorigenesis (i.e. upon inactivation of the APC gatekeeper). Their resistance to therapy and high regenerative potential makes them a critical driver of tumor plasticity and cancer recurrence. Specific Aims. We here propose to functionally characterize the OnFSCs in CRC. In Aim1, we will first generate phenotypic transcriptional reporters to track the dynamics of the LGR5+ CSCs and OnFSCs. Then, by coupling these reporters to a suicide gene (i.e. DTR), we aim to dissect the functional interplay between these different stem cell pools in CRC. In Aim2, we will use a multi-omic approach to determine the molecular drivers of the OnF state and phenotypic plasticity in CRC. In Aim3 we will perform targeted phenotypic screening to identify compounds able to selectively deplete OnFSCs. The top hits will be tested in combination with the current standard of care or LGR5+SC-targeting therapies11,12. We will validate our top drug combinations in vivo using both mouse tumoroids and human PDOs and/or PDXs. The aim is to determine a selective combination therapy superior to the current standard of care. Impact. The findings of these studies will uncover a new layer of complexity in CRC biology and will establish a mechanistic foundation for designing effective combination therapies with enduring impact on CRC treatment.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Across our lifetimes we have an infinite number of experiences that are stored in our memories. As we age our capacity to reuse this information to influence our future learning decreases, leading to impaired cognitive and behavioral flexibility. Cognitive decline is a major concern for an increasingly large and aging population, and these cognitive deficits will directly impact quality of life for countless individuals. It is essential to understand the neuronal coding deficits underlying cognitive decline to provide the necessary groundwork for developing therapeutic interventions that can spare people from cognitive decline. It has been postulated that the brain uses cognitive maps, or internal neural representations, to enable flexible behavior and relate items in our memory. The discovery of neurons in the hippocampus that fire action potentials in specific locations within an environment, termed place cells, provided initial support for the cognitive map hypothesis, as these cells create a neural representation of the environment. Both the engram and spatial navigation literatures have shown that distinct cognitive maps are used to encode two different environments, which would presumably reduce interference between maps during recall. However, recent work suggests that linking two distinct memories neuronally can enhance memory strength by sharing neural resources. Due to this linking, recall of one memory leads to a higher probability of recalling the other. This implies that some aspects of the two cognitive maps have increased in similarity and are no longer distinct. Others have hypothesized that increased similarity between cognitive maps may link experiences across environments and provide a mechanism to increase learning rates. Indeed, more recent evidence showed that neuronal representations in dCA1 became more similar as mice increased their speed of learning novel problems, suggesting that, under certain conditions, cognitive maps in the hippocampus may become increasingly similar, impacting learning. I will utilize in vivo calcium imaging with miniature microscopes as young adult and middle-aged mice navigate four distinct circular tracks for water rewards to ask the question of whether the similarity between neural representations underlying cognitive maps changes as mice increase their learning rates in novel environments. By using separate measurements of representational similarity, I can provide a detailed account of how the brain encodes new cognitive maps and whether the brain relates cognitive maps to alter learning rates. By using young adult and middle-aged mice, I can determine if these mechanisms change during aging. All results from this proposal have the capability to inform us on how the brain uses stored information to influence our future learning and, ultimately, how these mechanisms change during the early stages of cognitive decline.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The overall goal of this R21 application is to develop methods to predict how the presence of multiple subtypes of ventricular cardiomyocytes influences cardiac function. Recent studies using single-cell RNA sequencing (scRNAseq) have documented the presence of multiple cardiomyocyte subtypes, but these are described only in terms of gene expression patterns, and their functional consequences remain unclear. We will use a combination of approaches in this exploratory project to advance the linkage of gene expression to physiological function. We will first perform scRNAseq on purified cultures of CMs derived from induced pluripotent stem cells (iPSCs) to generate primary data about CM subtypes. Simulations with mechanistic mathematical models will then serve as a computational bridge linking gene expression and function, and physiological experiments to measure action potentials and intracellular [Ca2+] waveforms will test the predictions generated by the simulations. This R21 grant proposal is submitted in response to Notice of Special Interest NOT-HL-21-024, “Bold New Bioengineering Research for Heart, Lung, Blood and Sleep Disorders and Diseases.” The major advance of the planned research will be the development of computational tools, based on mechanistic models, that can link scRNAseq data to experimentally-testable predictions. The work will therefore not only provide new insight into cardiomyocyte subtypes, but will also deliver strategies and tools that can be applied in other contexts. This proposal combines the expertise of PI Eric Sobie, in cardiac physiology and mathematical modeling, with co-Is Ravi Iyengar, in systems biology and omics experiments, and Jens Hansen in computational analysis of omics data. The combined efforts of the investigators will generate new quantitative data and will yield new computational methods that can be applied broadly to understand cell subtypes in different contexts. To achieve the overall project goals, we will combine three complementary approaches, scRNAseq, mechanistic mathematical modeling, and physiological assays to test modeling predictions, as summarized in the following general goals: 1. Use scRNAseq to determine the characteristics and abundances of different subtypes in purified cultures of myocytes derived from induced pluripotent stem cells. 2. Perform simulations with mechanistic mathematical models to predict functional consequences of the presence of multiple cardiomyocyte subtypes. 3. Test predictions of the simulations in experiments measuring action potentials and intracellular calcium in purified cultures of cardiomyocytes.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Anal cancer represents approximately ~10% of all solid cancers among people with HIV (PWH) and is responsible for ~4% of all mortality in this group, representing a major morbidity burden in this population. Screening for anal precancerous lesions and treating these tumors has been proven to prevent invasive cancer and offers a clinical opportunity for addressing this emerging disease. However, standard screening tests (anal cytology and high-risk HPV testing) have very poor specificity leading to burdensome and invasive confirmatory testing. To address this we have identified genomic alterations that can identify anal high-grade squamous intraepithelial lesions (aHSIL), the immediate precursors of invasive cancer, with potentially greater accuracy and at lower cost than standard screening tests. However, the application of this approach for screening has not yet been investigated. The Specific Aims of this research proposal are (1) to compare the performance of a cytobrush-based, novel genomic test for the detection of aHSIL, to standard of care in a prospective cohort of people with HIV (PWH) at high risk of anal cancer and (2) to identify genomic alterations associated with aggressive aHSIL lesions and to develop a classifier the predicts lesions that are more likely to recur. To address these Aims, we will enroll a prospective cohort of 250 PWH for anal cancer screening and longitudinal data collection. This project will validate a genomic panel to identify aHSIL and compare it to standard of care screening as well as identify biomarkers associate with aHSIL recurrence. This application will also support Dr. Richard Silvera's career development into an independent patient-oriented investigator in the field of anal cancer among people with HIV. Dr. Silvera's career goal is to become a leader in developing novel strategies for the identification and prevention of anal cancers among PWH. The proposed career development plan integrates in-depth mentoring from a team of senior scientists and clinicians, advanced coursework in research design, analysis, cancer genomics, biomarker development, and experiential learning through the conduct of the proposed research in a highly supportive environment. The mentorship team, which includes independent investigators with extensive expertise in research of cancers among people with HIV as well as the genomics of cancer, will guide the candidate's research and career development. At conclusion of this project Dr. Silvera will be well positioned to become an independent physician-investigator studying biological and clinical issues related to anal cancers among people with HIV.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Beyond its role in conducting the air required for respiration, the airway epithelium plays a critical role in host defense through its physical and immunological properties. Specialized secretory cells present in the epithelium secrete airway mucins, which form a viscous hydrogel protecting the epithelial surface, and the coordinated beating of multiciliated cells propels this surface liquid and trapped foreign material away from the lower airways. TP63+ airway basal cells serve as the progenitor cell to replenish all other epithelial cell populations. Additional rare cell types with as-of-yet incompletely defined functions include FOXI1+ ionocytes and POU2F3+ tuft-like cells. All epithelial cell types constitute a selectively permeable physical barrier by forming cell-cell junctions with neighboring cells, such that an electrochemical gradient can be maintained across epithelial boundary. Besides these physical functions, airway epithelial cells participate in both innate and adaptive immune mechanisms through pathogen sensing, release of cytokines, and crosstalk with immune cells. The type I interferon (IFN-I) response is a broad-spectrum, first-line innate immune defense against viruses. While traditionally viewed as relatively uniform across cell types, preliminary data from our lab and collaborators show that IFN-I induces distinct gene expression programs across airway cell types, hinting at possible functional heterogeneity in the IFN-I response on a per-cell type basis. However, it is unclear whether these altered gene expression patterns and associated transcriptional regulators have functional significance in the context of respiratory virus infection. A diverse group of respiratory viruses infect the airway epithelium, including influenza viruses, rhinoviruses, “common-cold” coronaviruses, and SARS-CoV-2. These can be important triggers for acute exacerbations of chronic airway disease states. The proposed project will develop state-of-the-art CRISPRa/i functional genomics approaches to perturb IFN-induced regulators and effectors of innate immunity in a human, physiologically relevant model of the human airway epithelium. This will enable precise functional profiling of the IFN-I response in individual airway epithelial cell types. Altogether, this work will shed light on how the airway epithelium functions as an integrated unit to mediate host defense against respiratory viruses and strike a balance between effective host defense and immune homeostasis (using influenza A virus as a clinically relevant model respiratory pathogen), with the ultimate goal of developing novel host-directed antiviral therapeutic strategies. More generally, I anticipate the proposed experimental system developed here will be broadly valuable for precise genetic characterization of other airway disease states and their interaction with respiratory virus infection, including COPD, cystic fibrosis, and asthma. This project also presents a unique opportunity for training in both experimental and computational research approaches relevant to airway biology and respiratory virus infection.

NIH Research Projects · FY 2025 · 2024-09

Alzheimer’s disease (AD) affects more than 5.8 million elderly adults and is the most fatal dementia in the US. Patients diagnosed with AD present with cognitive decline including a loss in long-term memory. Perceptual memory is encoded on the synchronous firing of cells within a cortical network, and ensemble activity can be recalled with sensory cues, or it can be reactivated spontaneously during sleep. During sleep, the brain is placed in a quiescent state of slow-wave activity, where recently acquired memories are believed to spontaneously reactivate in order to consolidate into long-term storage. Human AD patients show memory deficits along with poor and fragmented sleep. Due to impairments in long term memory and sleep in AD patients and animal models, it is theorized that memory reactivation may also be impaired in AD. At this point memory reactivation has not been studied during sleep in AD. Animal studies where memory reactivation is artificially disrupted show memory deficits, and interestingly, animal research has demonstrated that spurring additional memory reactivation during sleep can improve memory consolidation. Interestingly, even the introduction of auditory stimulation to enhance slow-wave activity during sleep has been shown to aid the retention of new memories. In a recent study, stimulating slow- wave activity during sleep in human patients with mild cognitive impairments led to increases in memory retention. The overall goal of this proposal is to gauge memory reactivation in a mouse model of Alzheimer’s disease and to stimulate memory reactivation during sleep in AD mice to increase memory performance. I will use two-photon microscopy in parallel with recordings of electrophysiology to measure brain activity across wake and sleep. AD model animals will be imaged during sleep following a learning task to test the hypothesis that memory reactivation is impaired in an AD mouse model (Aim 1). In a separate set of experiments, the stimulation of virally delivered opsins will be used to spur additional memory reactivation in sleep following a learning task to test the hypothesis that spurring memory reactivation during sleep will improve memory in a mouse model of AD (Aim 2). The conclusion of this project will determine the specific alterations in memory reactivation during sleep contributing to memory deficits found in an AD mouse model, and identify the potential to modulate network activity during sleep to rescue memory deficits in a mouse model of AD. The completion of this preclinical research may influence early diagnosis of AD and lead to innovations of novel therapeutics for AD. Through the completion of this project, I will be exposed to training in optical techniques, electrophysiology, behavior, and sleep science in preclinical animal models to become an independent research scientist.

NIH Research Projects · FY 2026 · 2024-08

Up to one third of women, up to a billion people, experience the emergence or worsening of depressive and anxiety (mood) symptoms as their hormonal milieu shifts into perimenopause. Despite this proportion, we still do not understand what biological changes in perimenopause contribute to the increased risk of symptoms. Recent work suggests that perimenopause initiates a proinflammatory state and links both follicle-stimulating hormone (FSH) and acyl-ghrelin, two peripheral hormones that can cross the blood-brain barrier, to negative health outcomes. The roles of FSH and acyl-ghrelin extend beyond their canonical links to fertility and hunger, respectively. FSH receptors and ghrelin receptors are observed in many tissues and cells, including immune cells. FSH receptors are highly expressed in monocytes and FSH receptor activation stimulates proinflammatory cytokine production. Ghrelin receptors are expressed in the efferent vagus nerve, a well-known anti-inflammatory component of the autonomic nervous system, and acyl-ghrelin signaling increases vagal activity. Despite these observations, the hormonal mechanisms leading to mood symptoms remain elusive because the convergent hormonal pathways linking perimenopausal inflammation and mood symptoms have not been modeled. Our goal is to compare the time course of changes in FSH, estradiol, other reproductive hormones and acyl-ghrelin with the time course of changes in inflammation and mood. The study tests a mechanistic hypothesis in which elevated FSH and decreased acyl-ghrelin elevate proinflammatory markers through actions on monocytes and the autonomic nervous system, and thereby increase depressive and anxiety symptoms. This is a prospective longitudinal case-control study using a surgical menopause model, i.e. risk-reducing bilateral salpingo-oophorectomy (BSO). Surgical menopause reduces inter-individual variation in the onset and duration of perimenopause, thus facilitating our ability to measure associations between hormones, inflammation, and vagal activity pre- and post-menopause. We will recruit women seeking premenopausal risk-reducing BSO and age-matched premenopausal “controls” having non-BSO gynecological surgery. We will collect endocrine, immune and other measures pre-surgery, one week post-surgery and monthly post-surgery up to six months. Aim 1 will test the hypothesis that mood symptoms change with surgical menopause and that changes in hormones and proinflammatory markers relate to mood symptoms, specifically measuring plasma levels of FSH, estradiol, acyl-ghrelin, progesterone, liver-enriched antimicrobial peptide 2, cytokines and adipokines in basal plasma. Aim 2 will test the hypothesis that proinflammatory markers change with surgical menopause and that changes in acyl-ghrelin and vagal activity (vagal reflex, heart rate variability) relate to proinflammatory markers. Aim 3 (exploratory) will test the hypothesis that hormones relate to monocyte signaling changes with surgical menopause and that this signaling relates to proinflammatory markers. Mediation models will test for mechanistic effects across all aims.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic disorder characterized clinically by recurring episodes of pelvic pain and increased urination frequency, significantly impairing patients' quality of life. Irritable bowel syndrome (IBS) is a multifactorial disorder characterized by abdominal pain and altered bowel habits, as well as other somatic, visceral, and psychiatric comorbidities. Despite distinct origins, recent studies have shown a strong overlap between both syndromes and this pathological co-occurrence is believed to be responsible for the comorbidity of a number of lower urinary tract and colonic disorders, including, but not limited to, IBS, inflammatory bowel disease (IBD), overactive bladder (OAB), and IC/BPS. There is an unmet medical need in terms of effective diagnostics and treatment. One of the challenges is a lack of understanding of the molecular and cellular mechanisms underlying the pathogenesis of neurogenic bladder pain. Moreover, how bladder inflammation promotes colon sensitization is poorly understood. Pilot studies showed that MrgprB2 is required for bladder pain and sensitization of colon to mechanical stimulation induced by intravesical instillation of LL-37. These observations led us to frame the hypothesis that MrgprB2+ mast cell (MC)-TRPV1+ nociceptor clusters mediate bladder pain and inter-organ sensitization between the bladder and colon. In this grant proposal we will: 1) Use chemogenetic inhibition of mast cells and virally mediated ablation of TRPV1+ bladder-innervating nociceptors to investigate in vivo functions of the MrgprB2+ MC-TRPV1+ nociceptor clusters in MrgprB2-dependent bladder pain; use humanized mouse model with targeted expression of MrgprX2 in MCs to test if human MrgprX2 also impacts bladder pain; use TRPV1+ nociceptor specific Piezo2 cKO mice and virally mediated ablation of Piezo2 function in bladder-innervating nociceptors to determine if nociceptor-expressed mechanosensitive Piezo2 channels mediate bladder pain induced by various irritants including LL-37, compound 48/80, or CYP. 2) Focus on differentiating visceral nociceptors innervating both the bladder and colon and use unique chemical- and viral-mediated approaches to either ablate or chemogenetically inhibit these distinct nociceptors to determine if they are required for the generation of colon pain induced by bladder inflammation caused by LL-37, compound 48/80, or CYP; test if MC-derived LTC4-neuronal CysLTR2 axis contributes to bladder pain and mediates inter-organ cross sensitization in the colon. Our findings will provide cellular and molecular basis for bladder pain and bladder-colon cross sensitization mediated by the MrgprB2+ mast cell-TRPV1+ nociceptor clusters and will undoubtedly lead to new therapeutic approaches for treating visceral pain.

NIH Research Projects · FY 2024 · 2024-08

Summary: Human cytomegalovirus (CMV) is a β-herpes virus with high seroprevalence rates of 60-90% within the population that can spread through bodily fluids, organ transplants, and from mother to fetus via the placenta. Virus proliferation significantly increases the morbidity and mortality of immunocompromised individuals, such as newborns, organ transplant recipients, AIDS patients, and the elderly. Approximately 30,000 solid organ transplant and 23,000 bone marrow transplants operations are performed in the U.S. every year. CMV is the leading cause of birth defects, affecting ~1% of newborns giving rise to ~30,000 new cases of CMV infection reported annually in the US. While several drugs have been approved by the FDA for the treatment of CMV infections, including ganciclovir, foscarnet, letermovir, and recently maribavir, these compounds were found to exhibit high frequencies of drug resistance and severe side effects, including bone marrow toxicity, gastrointestinal disruption, and nephrotoxicity. Given the large number of patient populations at risk for CMV- associated diseases and the estimated cost to treat CMV in the US ($4.4 billion/year by the National Academy of Sciences), novel therapeutic strategies to treat CMV-associated diseases is needed. The development of novel therapeutics that target different steps of the viral life cycle to limit virus propagation and dissemination would provide therapeutic for treating CMV-related diseases. We developed a high-content screening assay using a CMV AD169 reporter virus to screen >112,000 compounds in collaboration with Microbiotix to identify inhibitors that block the early stage of infection. One compound, MBXC-4302, is a N-arylpyrimidinamine (NAPA) that exhibited potent anti-CMV infection activity (IC50 ~3 µM), limited cytotoxicity (CC50 >100µM), favorable in vitro ADME properties, and a responsive structure activity relationship (SAR). Commercial analogs of MBXC-4302 identified MBXC-4992 with an improved selective index. Our general hypothesis is that the NAPA compounds represent novel CMV entry inhibitors that can be developed into effective CMV therapeutics. To evaluate our hypothesis, we plan to complete the following Aims: 1) Characterize the mechanism of action and broad inhibition of the NAPA compounds; 2) Optimize the NAPA series through SAR- driven analog generation; 3) Evaluate the ability of NAPA compounds to limit virus proliferation and dissemination in diverse in vitro models; and 4) Evaluate pharmacokinetics, tolerability, and efficacy of prioritized NAPA analogs in dissemination studies in vivo. We plan to develop several NAPA analogs as effective CMV therapeutics that inhibit CMV dissemination as a single agent or in combination with FDA approved CMV drugs.

NIH Research Projects · FY 2026 · 2024-08

Project Summary To feel, remember and act in a social environment, incoming social stimuli must be processed and represented. Recent advances have revealed several routes of social information flow which converge on the dorsal (d)CA2. Ensembles encoding individual identity have also been found in the dCA2, but how diverse social information is integrated to form coherent social representations remains elusive. Studies in human participants have provided early evidence that the hippocampus tracks two social features, familiarity and hierarchy, jointly. However, it is poorly understood how neural ensembles encode social characteristics and how incoming information instruct the formation of social representations based on those characteristics. To address this major knowledge gap, our overall objective is to bridge circuit and population coding mechanisms to determine the neural basis of social information processing. As familiarity and hierarchy are two factors by which social relationships are evaluated across species, we reason that dissociating their circuit and population coding mechanisms will reveal how social information is structured in the brain. Our preliminary data suggest that the dCA2 is functionally relevant for hierarchy and familiarity behaviors and its activity scales with rank distance. Given that the entorhinal cortex shows grid-like activity in rank representation, we hypothesize that the dCA2 may display place cell-like dynamics to signal familiarity and hierarchy distance on an abstract social cognitive map. We will test this hypothesis in Aim 1 with bulk (fiber photometry) and single-cell imaging (Miniscope) of the dCA2 in response to varied familiarity or hierarchy stimuli. Using the imaging data, we will evaluate predictive models of coding strategy with linear classifiers. In Aim 2 we will assess how social presentations of familiarity and hierarchy arise. We have preliminary evidence that distinct neuromodulator activity in the dCA2 correlates with rank distance. As neuromodulators are mainly released into the dCA2 by the paraventricular nucleus of the hypothalamus (PVH), we hypothesize that specific neuromodulators released by the PVH differentially impacts the geometry of the social characteristics. Thus, we will use novel neuromodulator sensors to monitor their dynamics and manipulate their signaling during Miniscope recordings. Finally, we will test their functional relevance by inhibiting their release from the PVH during hierarchy and familiarity behaviors. The approach detailed in this proposal is highly innovative because it leverages new technological advances (novel neuromodulator sensors) and integrates cutting-edge techniques (manipulation of neuromodulator signaling with Miniscope imaging and computational modeling) to address a major knowledge gap in social cognition. As no unified theories of circuit and population coding mechanisms for social information processing exist, the proposed research is highly significant. A mechanistic understanding of how neuromodulatory inputs regulate neural population coding will provide druggable targets, which have the potential to transform therapeutic treatment of social cognitive deficiencies in neuropsychiatric disorders.