Vanderbilt University

NSF Awards · FY 2024 · 2024-06

This project explores a proof-of-concept and feasibility evaluation to inform the future development of a centralized data repository to support the privacy research community. The repository will enable tracking and systematic study of privacy harms. Current incident reporting systems are designed to track the occurrence of large-scale data breaches, but there is currently no centralized reporting system to effectively track other types of privacy violations (e.g., online harassment, cyber abuse) that negatively impact end-users. Without access to this information, it is difficult to quantify / qualify how and to what extent different online platforms propagate privacy breaches, as well as how to redesign such systems to be more secure and trustworthy. Therefore, this planning effort aims to (1) solicit the opinions of privacy experts on the design of the repository; (2) prototype the repository and solicit feedback from experts piloting it; and (3) build on these learnings to develop a plan to develop a centralized privacy incident repository. This will ultimately enable researchers to work together to (1) identify and prioritize privacy harms and the factors associated with the incidents; (2) understand how various populations are impacted by these harms; and (3) develop and evaluate potential interventions. This repository is envisioned to support the protection of vulnerable end-users who are disproportionately threatened and harmed by digital privacy violations, addressing the recent R&D budget priority from the White House and the Office of Science and Technology Policy focused on reducing inequities. By identifying evolving privacy risks, we also work towards two other budget priorities -- advancing trustworthy AI technology and maintaining global security and stability. 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.

NSF Awards · FY 2024 · 2024-06

While there has been extensive research on the barriers Black and brown students face as they strive to participate in engineering education and the workforce, there is less scholarship on solutions for addressing this complex challenge. One reason for this is because the scholarship on how change happens in engineering education tends to focus on course content and classroom instruction. Unfortunately, such findings do not easily lend themselves to value-laden, systemic issues like diversity, equity, and inclusion (DEI). Fortunately, some Colleges of Engineering (COEs) throughout the U.S. have adopted change strategies that have resulted in consistently being named among the top-ten producers of Black and brown engineers. This project is motivated by a desire to learn from and follow their example. This CAREER project will disrupt the status quo regarding who gets to be an engineer by investigating five COEs that have significantly changed the face of engineering over the last 20 years. This project will: (1) Advance our understanding of the change strategies that exemplary COEs have used to improve Black and brown students’ access to engineering education and careers; (2) Identify evidence-based models for broadening participation of underrepresented racial/ethnic groups in engineering; and (3) Set COEs on a path to parity, such that the student body demographics in COEs across the country reflect the racial/ethnic makeup of the nation. Using Kotter’s Leading Change Model and Acker’s Inequality Regimes as a framework, this multi-case study will investigate how exemplary COEs envisioned, implemented, and institutionalized changes that influenced Black and brown students’ access to engineering. The five COEs that will be investigated are: Florida International University, Morgan State University, University of Central Florida, University of Maryland-Baltimore County, and University of Maryland-College Park. Given variations in the types of universities included in the research design, comparing and contrasting insights that emerge from each case will enable the PI to understand the conditions for change. The use of a research study design that relies on both qualitative and quantitative data will produce complementary forms of evidence on what promotes and impedes progress in this context. The research outcomes will include: (1) impact narratives that document concrete examples of how to expand who gets to be an engineer; and (2) a model for broadening participation informed by a cross-case analysis of these exemplars. Furthermore, this timely work focuses on the need to leverage talent from every demographic to diversify the engineering workforce and improve the lived experiences of minoritized groups. The educational outcomes will include: an Impact Playbook that translates the research into actionable strategies; a graduate course for future engineering faculty designed around each of the cases; a townhall discussion among associate professors; sharing insights with ASEE’s Engineering Deans Council; and a partnership with Virginia Tech’s (VT) College of Engineering and College of Science to build capacity among its leaders to envision and enact sustainable changes that promote DEI on VT’s campus. This CAREER project has the potential to reshape how COEs approach their DEI efforts, and increase the likelihood of long-term success. The proposed activities are designed to foster a network of STEM leaders motivated to envision and enact sustainable, scalable changes that expand who gets to be an engineer at their institution. 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 · 2024-05

PROJECT SUMMARY Granule cells (GCs) constitute over 95% of the cerebellar volume. They receive and integrate sensory, motor, and non-sensorimotor signals to fine-tune motor behaviors and cognitive tasks. GCs are generated from transiently proliferating granule cell precursors (GCPs) over a long time extending from early embryonic period until first postnatal year in human. Accordingly, cerebellar hypoplasia is one of the most common brain complications in premature infants with poor developmental outcomes. We have very limited basic knowledge of how GC lineage is established. Our long term goals are to elucidate the regulatory mechanisms of GC lineage development, and to understand how different risk factors cause cerebellar hypoplasia. A master regulator of GCP development is the bHLH transcription factor Atoh1 that maintains the GCP fate through activation of its own expression. This autoregulatory feedback loop is further supported by a cell cycle regulator Ccnd1 that stabilizes Atoh1 protein from degradation. However, it remains unclear as to how Atoh1 and Ccnd1 expressions are terminated to enable timely progression from GCPs to GCs. Our preliminary data suggest that Sin3A, a component of histone deacetylase (Hdac)–containing transcriptional corepressor complex, is essential for GCP differentiation by epigenetically silencing Atoh1 expression. We have also identified Insm1, a zinc-finger transcription factor, as a potential partner of the Sin3A-Hdac complex that inhibits Atoh1 and Ccnd1 expression. Based on these and other preliminary observations, we propose the novel hypothesis that the Sin3a/Hdac/Insm1 complex epigenetically represses Atoh1 and Ccnd1 expression, thereby integrating transcriptional and posttranslational signals to regulate GCP fate and cell cycle progression. This hypothesis will be tested by establishing (1) Sin3A function during GC lineage development, and (2) the role of Insm1 in GCP differentiation.

NIH Research Projects · FY 2026 · 2024-05

Project Summary/Abstract: The objective of this proposal is to create a new surgical robot to enable women who currently face the life- long consequences of hysterectomy to have minimally invasive, uterine-sparing interventions. The robot will deliver needle-sized instruments through an endoscope, enabling independent tissue manipulation, electrosur- gical probe movement, and visualization, facilitating more precise, accurate, and efficient intrauterine surgery. Clinical significance comes from the fact that over 50,000 women per year will lose their uterus to fibroids that elite surgeons have demonstrated can be removed endoscopically. Instrument dexterity limitations prevent typical surgeons from offering uterine-sparing endoscopy to these women. This results in only 1 in 10 women benefiting from the minimally invasive endoscopic approach while 2 in 3 face the lifelong negative consequences of hysterectomy. Our innovation is to provide the surgeon with two miniature robotic instruments delivered through the endo- scope, to enhance dexterity and enable two-handed retraction and resection. These instruments are made from telescoping, curved, elastic tubes. By axially rotating and telescopically extending the tubes, our robot will provide the surgeon with two tentacle-like instruments small enough to be delivered through standard-sized endoscopes. These instruments, combined with a variable view angle optic, will enable lateral dexterity and visualization, countertraction, and accurate and efficient tissue resection. Our approach consists of three Specific Aims. Aim 1 addresses the design of our new instruments, in con- junction with the variable view angles provided by our optic, and endowing them with bipolar electrosurgery capability. Aim 2 focuses on the design of a novel touchscreen-based user interface that enables the surgeon to simultaneously control the endoscope and the instruments delivered through it. Aim 3 consists of ex vivo and in vivo animal studies to demonstrate the robot’s ability to reach everywhere in the uterus, to enable surgeons to perform the procedure who otherwise could not, and to resect FIGO type 0, 1, and 2 fibroids of various sizes from all relevant locations (the fundal, anterior, posterior, lateral-left, and lateral-right zones) of the uterus efficiently. The endpoint of this R01 is a device that has been fully validated in established clinical training scenarios and live animals, setting the stage for clinical translation after the successful completion of this R01.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Cardiovascular disease (CVD) is the leading cause of death globally. The root cause of CVD is a chronic inflammatory disease of the arterial wall called atherosclerosis. A critical determinant of atherosclerosis is macrophage inflammatory phenotype. Within atherosclerosis, anti-inflammatory pathways are dysfunctional, which results in pro-inflammatory stimuli driving macrophage phenotype. We have identified Paired immunoglobulin-like receptor B (PirB) as a novel inhibitor of macrophage-mediated inflammation by facilitating apoptotic cell digestion and promoting mitochondrial function. The aims of this proposal will determine the mechanisms by which PirB inhibits inflammation through phagosomal and mitochondrial signaling pathways. We will use macrophages containing a lysosomal reporter to track phagosome maturation and cargo fate. Additionally, we will dissect the molecular mechanism by which PirB regulates mitochondrial metabolic function and signaling.

NIH Research Projects · FY 2026 · 2024-05

The long-term goal of the PI’s research program is to illuminate the structure, dynamics and mechanistic principles that underpin active efflux of solutes, often of impressive size such as protein domains, across cell membranes. Clinical multidrug resistance (MDR) in the treatment of bacterial and fungal infections, and chemotherapy of neoplasms can be associated with overexpression of membrane-embedded efflux pumps, collectively referred to as MDR transporters, that selectively extrude cytotoxic molecules from the cell. MDR transporters harness the free energy of ATP hydrolysis or that stored in electrochemical gradients to power a conformational cycle that drives the energetically uphill vectorial translocation of substrates. The cycle entails the energy-coupled isomerization of the transporter between multiple intermediates thereby executing alternating access of the substrate binding site. Defining the structural elements mediating alternating access and decoding the mechanism of energy conversion in a lipid bilayer environment are central questions in the field and critical for elucidating transport mechanisms. This MIRA proposal will continue support of two established, productive research programs focused on addressing these questions for ATP binding cassette (ABC) and Multidrug and Toxin Extrusion (MATE) transporters. Our innovative experimental blueprint capitalizes on recent transformational advances in machine learning protein structure prediction, state of the art electron paramagnetic spectroscopy (EPR) tools in the context of high resolution cryoEM structures. Project 1, motivated and grounded in the contribution of the PI’s group, seeks to decipher ion-substrate coupling, to define conserved and divergent elements of alternating access, and to reveal specific transporter-lipids interactions that shape the energy landscape of conformational changes in MATE transporters. Project 2 will expand a long-standing investigation of energy transduction and alternating access ABC efflux transporters in three archetypes that represent a spectrum of energy conversion and substrate size. The two projects will illuminate mechanistic principles for families of transporters implicated in the phenomena of drug resistance and basic bacterial defense strategies.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY/ABSTRACT Approximately 40% of patients with epilepsy are resistant to drug treatment. These patients may benefit from targeted surgical interventions, such as surgical resection or neurostimulation of the seizure onset zone (SOZ). Patients being considered for surgical intervention undergo a series of noninvasive diagnostic tests to localize their SOZ. If these noninvasive tests cannot delineate the SOZ clearly enough for surgical intervention, patients undergo intracranial electroencephalography (iEEG) to more accurately localize their SOZ. Yet, even in patients who undergo iEEG, SOZ localization and subsequent surgical treatment only leads to seizure freedom in 47- 68% of patients, which can be partially attributed to SOZ mis-localization. Thus, there is a need for more accurate noninvasive SOZ localization methods to guide iEEG implantation and surgical treatment for improvement of seizure outcomes after surgical intervention. Resting-state, interictal (between seizure) functional magnetic resonance imaging (fMRI) noninvasively measures fluctuations in blood oxygenation, an indirect measure of brain activity. In this proposal, we aim to develop two novel interictal fMRI SOZ localization methods that improve upon current noninvasive methods by leveraging interictal physiological abnormalities of the SOZ. We will develop these methods in patients with temporal lobe epilepsy as a well-characterized model of focal epilepsy. Interictal epileptic spikes, large amplitude electrical events, are detected clinically with noninvasive scalp EEG to localize the SOZ. However, clinical interictal spike localization methods are limited by low sensitivity to detect spikes and either poor spatial resolution or specialized hardware and extensive preprocessing. There is evidence that interictal epileptic spikes induce specific dynamic fMRI connectivity patterns, therefore, here we propose to develop a method to detect and localize interictal epileptic spikes with dynamic fMRI connectivity (Aim 1). We hypothesize that interictal epileptic spikes identified on scalp EEG induce dynamic fMRI connectivity patterns that could be used to detect and localize spikes with fMRI alone. Fluorodeoxyglucose (FDG) positron emission tomography (PET) is used clinically to image the glucose hypometabolism of epileptic regions, however, this method has limited utility for SOZ localization due to the moderate specificity of hypometabolism to the SOZ. We aim to improve the specificity of FDG-PET SOZ localization by combining it with fMRI (Aim 2). We hypothesize that the SOZ has atypical physiological uncoupling of interictal metabolic (FDG-PET) and hemodynamic (fMRI) activity that could be used for localization. If successful, these studies will provide more accurate and specific noninvasive SOZ localization methods that could be integrated into presurgical noninvasive testing to guide both iEEG implantation and surgical intervention, ultimately improving seizure outcomes after surgical intervention. Moreover, these studies will investigate the fMRI correlates of interictal electrophysiological and metabolic abnormalities in focal epilepsy which may provide insights into the pathophysiological underpinnings of fMRI alterations previously reported in epilepsy.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY Gastric cancer is currently the fourth leading cause of cancer deaths worldwide and is diagnosed in approximately one million people per year. The most common type of gastric cancer is intestinal-type, which develops through a series of metaplastic and dysplastic lesions of the stomach lining beginning with pyloric metaplasia. The hallmark of pyloric metaplasia is the transdifferentiation of gastric chief cells into spasmolytic polypeptide-expressing metaplastic (SPEM) cells. While gastric carcinogenesis is histologically well defined, the molecular drivers of the cellular and morphological changes that occur in each of these stages have not yet been discovered. Transcription factors are proteins that control cell fate via gene expression during development and in adult progenitor cell differentiation. To determine if transcription factors also control cell fate in metaplastic processes in the stomach, my lab probed for the expression of select factors in carcinogenic gastric tissue and found a strong upregulation of the master transcription factor SOX9 in SPEM cells. To follow up on this finding, I characterized two novel transgenic mouse models of pyloric metaplasia with Sox9 knock- out in chief cells and found that SPEM cell formation and subsequent carcinogenesis was severely disrupted. I hypothesize that SOX9 is necessary for chief cells to transdifferentiate into SPEM cells which are essential for carcinogenesis. To understand the significance of SOX9 in SPEM cells and carcinogenesis, I propose three specific aims. First, I will investigate the functional role of SOX9 by performing an in vitro experiment using organoids from Sox9 knock-out tissues to determine if SOX9 is required for chief cells to transdifferentiate. I will also examine whether Sox9 knock-out chief cells are able to stimulate to transdifferentiate into SPEM cells. Second, I will perform transcriptomic profiling to identify direct downstream genes of SOX9 to elucidate its transcriptional mechanism in SPEM cell identify and function. Finally, I will investigate the success of microenvironment recruitment which is essential to the progression of carcinogenesis without SOX9 upregulation in chief cells. This will elucidate a biological mechanism by which SPEM cells drive carcinogenesis. Together, these data will provide functional and transcriptional mechanisms of SOX9 in SPEM cells during gastric cancer development. This research will move the field forward by uncovering the molecular mechanisms of gastric carcinogenesis and will provide potential therapeutic targets for preventing gastric cancer development.

NIH Research Projects · FY 2025 · 2024-04

Abstract There are reported sex differences in both the propensity to develop nicotine addiction and the efficacy of treatment interventions pointing to women as a particularly vulnerable population. Thus, understanding how sex differences in nicotine effects on the brain arise is a critical unanswered question in addiction neuroscience. Dopamine signaling within the nucleus accumbens is involved in initiation and maintenance of nicotine addiction, and dopamine release is sensitive to heavy modulation by acetylcholine through nicotinic receptors located on dopamine axons. Acetylcholine modulation of dopamine release at distal axon terminals is discussed extensively, and better understanding this interaction has driven experimentation across fields - critically, we find that this interaction is not present in adult intact females. However, our preliminary data show that this effect is rescued in females after ovariectomy, suggesting that ovarian hormones play a critical role in modulating acetylcholine effects on dopamine release; the goal of this proposal is to define where this modulation occurs and how this influences nicotine effects on this system. This proposal is guided by the overarching hypothesis that estradiol acts acutely through estrogen receptors located within acetylcholine interneurons in the nucleus accumbens core to strengthen acetylcholine effects on dopamine release. However, in intact females, long-term estrogen modulation of this interaction leads to an inability of modulators – such as nicotine – to further enhance dopamine release. This is supported by robust preliminary data showing that in males and ovariectomized females estradiol effects on dopamine release can be blocked via nicotinic receptor antagonism. In intact females, estradiol effects on dopamine release are blunted and insensitive to nicotinic receptor agonists and antagonists. This fellowship application describes a robust training plan to investigate the microcircuitry of how ovarian hormones mediate dopamine-acetylcholine interactions and the role that this plays in attributing motivational value to nicotine-associated cues. Aim 1 will define the estrogen receptor subtypes contributing to estrogen effects on acetylcholine-dopamine interactions. Aim 2 will determine how estrogen receptors regulate acetylcholine release using optical acetylcholine sensors. Aim 3 will use genetic approaches to knock out estrogen receptors from acetylcholine interneurons and determine how this affects the ability of dopamine to respond to nicotine and nicotine-associated cues in operant tasks. This plan expands on my current electrophysiology expertise to train me on optogenetic tools and optical imaging and gives me the career development opportunities necessary to propel me to a successful placement in a tenure-track position at a respected institution. Together, the results of this project will critically inform the addiction field on the effects of ovarian hormones on dopamine interactions in the nucleus accumbens core, providing new directions for more effective treatment of addiction. .

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY/ABSTRACT Evidence suggests that cocaine use, notably during critical neurodevelopmental periods like adolescence, is associated with significant and persistent impairments in cognitive processes like working memory and cognitive flexibility. Cognitive symptoms are associated with loss of GABAergic inhibitory transmission in the prefrontal cortex (PFC). Our preliminary data demonstrates that adolescent cocaine exposure induces persistent deficits in GABAergic somatostatin-expressing interneurons (SST-INs) in the PFC and cognitive performance in adulthood. Thus, enhancing cortical inhibition represents a novel therapeutic approach to alleviate cognitive deficits associated with adolescent cocaine exposure and cocaine use disorder (CUD). This therapeutic goal may be achieved via activation of the mGlu1 subtype of metabotropic glutamate (mGlu) receptor. Leveraging highly selective mGlu1 positive allosteric modulators (PAMs), we recently found that mGlu1 positively regulates cortical inhibition via actions on SST-INs and improves cognitive performance. These findings led us to our central hypothesis that the ability of mGlu1 PAMs to reverse deficits in cortical inhibition and cognitive impairment associated with adolescent cocaine exposure is likely driven by actions on mGlu1 receptors located on SST-INs in the PFC. The present proposal will use cutting-edge transgenic mouse lines and pharmacological tools to elucidate brain region- and cell type-specific mechanisms underlying mGlu1 regulation of cortical inhibition and cognitive function and their implications following adolescent cocaine exposure. We will test this hypothesis in three specific aims. Aim 1 (K99): To gain expertise in whole-cell patch-clamp electrophysiology to test the hypothesis that mGlu1 regulates PFC inhibitory transmission via actions on SST-INs. Aim 2 (K99): To develop expertise in in vivo fiber photometry to test the hypothesis that mGlu1-mediated activation of PFC SST-INs facilitates working memory performance. Aim 3 (R00): To apply whole-cell patch-clamp electrophysiology and in vivo fiber photometry to test the hypothesis that mGlu1 modulation can ameliorate adolescent cocaine-induced pathophysiology through actions on PFC SST-INs. Together, the proposed studies will expand our current understanding of the critical role of SST-INs in cognitive processes impaired by adolescent cocaine use and evaluate the therapeutic potential of mGlu1 PAMs in mitigating these cocaine-induced deficits. I will gain extensive training in whole-cell patch-clamp electrophysiology and in vivo cell type-specific Ca2+ imaging during touchscreen-based cognitive testing. Training in these techniques will allow me to apply my interests in behavioral and receptor pharmacology to unanswered questions about how drug exposure during critical neurodevelopmental periods manifest in physiological and behavioral consequences of substance use disorders.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY With age and exposure to genotoxic stress, tissues throughout the body acquire somatic mutations. In the blood and bone marrow, somatic mutations are frequently found in leukemia-associated genes in subsets of hematopoietic cells even in the absence of hematologic cancer. This phenomenon has been termed Clonal Hematopoiesis of Indeterminate Potential (CHIP) because while many people with such mutations will have no known clinical impact, the presence of these clonal populations is associated with increased risk or poor outcomes in diseases ranging from hematologic malignancy and cardiovascular disease to osteoporosis and COVID-19. While CHIP affects at least 10% of people over age 70, it impacts as many as 25% of patients with solid tumors, likely driven by the genotoxic stress of chemotherapy and radiation. Recent studies have shown that CHIP may result in aberrant inflammatory programming, particularly in myeloid lineages, and that immune cells with CHIP mutations can infiltrate the tumor microenvironment. Moreover, research suggests that CHIP may be associated with worse overall survival among solid tumor patients. Given the importance of the tumor immune microenvironment and the increased burden of CHIP in these populations, there is a fundamental need to examine the interplay between CHIP and solid malignancies and to explore how to clinically manage solid tumor patients with CHIP. This proposal seeks to use clinical sequencing data, patient samples, mouse models, and ex vivo experiments to determine the impact of CHIP on solid tumor outcomes and immune microenvironment using triple negative breast cancer (TNBC) as a model system. TNBC is an aggressive subtype of breast cancer that lacks expression of estrogen receptor, progesterone receptor, and HER2. Importantly, TNBC often has a prominent immune cell infiltrate that is prognostic: the presence of lymphocytes is associated with favorable outcomes, while the presence of tumor-associated macrophages is a negative prognostic indicator. Aim 1 will leverage multiple large biobanks, prospectively collected patient specimens, and a novel mouse model to define CHIP as a prognostic biomarker for TNBC outcomes, including overall survival and response to therapy. Aim 2 will harness both patient-derived immune cells and mouse models to determine how myeloid cells with CHIP mutations interact with TNBC cells. Techniques to be utilized include histopathology, flow cytometry and immunophenotyping, gene expression analysis, and functional immune cell assays. Completion of this work will provide a comprehensive training vehicle for this fellowship and will simultaneously yield new insights into clinical management of solid tumor patients with CHIP, suggesting novel therapeutic approaches for these patients. These studies will also provide a greater understanding of how dysregulated myeloid cell signaling in CHIP clones leads to differential interaction in the tumor microenvironment.

NIH Research Projects · FY 2026 · 2024-04

Title: Glial roles in experience-dependent critical period remodeling Summary We propose glia actively prune brain circuits to optimize connectivity based on early-life sensory experience. Drosophila approaches are used to identify molecular mechanisms of activity-dependent glial pruning restricted to this short critical period. We propose these glial mechanisms go awry in several newly-linked disease states of intellectual and autism spectrum disorders, including Fragile X syndrome (FXS), Noonan syndrome (NS), LEOPARD syndrome (i.e. NS with Multiple Lentigines; NSML), and Neurobeachin (NBEA) associated autism spectrum disorder (ASD). Our plan is to test the role of glia in experience-dependent circuit pruning in normal and disease states, and to order glial mechanisms of recruitment, infiltration, engulfment and phagocytosis. We employ targeted CRISPR knockout, conditional gene manipulations, and transgenic brain circuit connectivity mapping to dissect neuron-to-glia signaling and glia function in this sensory experience-dependent remodeling. We use timed olfactory cues to activate odorant receptor neurons, downstream projection neurons, and central learning/memory center Kenyon cells, to test glial phagocytosis activity during and following the critical period. In Aim 1, we block glial phagocytic function at multiple levels to test experience-dependent connectivity pruning throughout this defined brain circuitry. We use transgenic single neuron synaptic labeling to visualize glial phagocytosis via transmission electron microscopy. To test downstream activity-dependent mechanisms, we use both excitatory and inhibitory optogenetic tools, as well as transgenic blockage of neurotransmission, in hierarchical circuit studies of synaptic connectivity is sequential brain neuropils. This aim systematically tests glial pruning in normal juvenile brains. In Aim 2, we dissect glia-specific Fragile X Mental Retardation Protein (FMRP) roles in experience-dependent brain circuit pruning. We assay FMRP requirements in glial infiltration phagocytosis using combined light microscopy and ultrastructural imaging. We test the FMRP-dependent neuron-to-glia signaling mechanisms of glial recruitment and phagocytosis during experience-dependent circuit remodeling. This work distinguishes FMRP roles within glia and neurons to understand FXS disease model impairments in critical period brain circuit remodeling. In Aim 3, we test the roles of FMRP translational targets, and consequent regulation of PKA/ERK signaling pathways. We test roles of 1) the direct FMRP mRNA target Rugose/NBEA causative in autism spectrum disorder, which acts as a regulatory PKA anchor, and 2) the direct FMRP mRNA target Corkscrew/SHP2 causative in the two Noonan syndromes of intellectual disability, which is an ERK pathway regulatory phosphatase. We use separation of phases-based activity reporter of kinase (SPARK) biosensors to image experience-dependent PKA/ERK signaling during the early-life critical period. Taken together, this research program dissects gene-environment interactions in normal critical period brain circuit pruning by glial phagocytes, with translational links to new molecular mechanism intersections between Fragile X syndrome, two related Noonan syndromes, and Neurobeachin-associated autism spectrum disorder.

NIH Research Projects · FY 2025 · 2024-04

PROJECT ABSTRACT Spinal cord injuries (SCI) initiate a cascade of complex physiological and molecular mechanisms at and around the site of injury. Beyond the initial damage, secondary and chronic neuropathological effects triggered by neuroinflammation and molecular changes at the injury site can severely impact spinal cord regeneration. Some of the mechanisms of interest include glutamate excitotoxicity, demyelination, glial scar formation, toxic free radical accumulation, and pro-inflammatory cytokine release. These secondary and chronic pathological effects can severely impede spinal cord regeneration through inhibition of axonal regrowth and progressive cell death at the injury site. Multi-parametric MRI (mpMRI) provides an array of contrasts sensitive to changes that occur in SCI. Quantitative magnetization transfer (qMT) imaging characterizes myelin concentration changes through measurements of immobile macromolecular content. Diffusion tensor imaging (DTI) provides complementary structural information by evaluating spinal cord axonal tract integrity post-injury. Resting-state functional MRI (rsfMRI) reports the integrity of gray matter resting state functional networks that are disrupted post-injury. Chemical exchange saturation transfer (CEST) and Nuclear Overhauser Enhancement (NOE) imaging generates Z-spectra that reflect changes in the concentrations and/or exchange rates of specific metabolites and macromolecules, providing high resolution molecular information from the SCI region. In addition, PET imaging using radiotracers that bind specifically to the translocator protein (TSPO) will provide three-dimensional spatial maps of neuroinflammatory activity in the injured spine and can validate the interpretation of mpMRI measures. We hypothesize that using mpMRI, we can detect and quantify relevant structural, functional, and molecular changes in the spinal cord, longitudinally over time, in a rat injury model, and that imaging metrics can be used as biomarkers of the effects of therapeutic interventions. These biomarkers will correlate with animal functional recovery and can be used to evaluate the efficacy of SCI treatments. We will evaluate the validity of these hypotheses in the proposed study. First, we will optimize and implement CEST and NOE MR sequence protocols for imaging SCI rats. Next, we will combine mpMRI imaging modalities to measure the sensitivity of mpMRI for quantifying SCI severity and recovery and for evaluating the efficacy of SCI treatments, in a treatment study using the neuroprotective drug Riluzole. Finally, we will confirm the structural, molecular, and behavioral basis of the mpMRI imaging metrics through TSPO PET imaging of neuroinflammation, motor and somatosensory behavior testing, and histological analysis. The proposed studies are significant because they will develop, validate, and implement innovative mpMRI methods that can evaluate and track various neuropathological changes that accompany recovery after SCIs in a clinically relevant animal model. These methods may then be used to assess novel SCI interventions and treatments, and to provide comprehensive information on structural, functional, and molecular changes in SCIs over time.

NIH Research Projects · FY 2026 · 2024-03

Project Summary This application is in response to Funding Opportunity Announcement PAR-21-034 a five day course entitled” Isotope Tracers in Metabolic Research: Principles and Practice in Kinetic Analysis”. The course faculty will present state of the art methods for using both radioactive and stable isotopes to investigate whole body and organ metabolism in vivo and intracellular flux rates and pathway regulation in vivo and in vitro. The basic aspects of modeling will be considered, as well as specific applications to the study of carbohydrate, fat, protein metabolism and energy balance. Theoretical and practical matters related to sample analysis by mass spectrometry and NMR will be discussed, including detailed numerical examples of calculations involved in determining isotopic enrichment and basic kinetic parameters. Advanced lectures will discuss in more detail the use of positional and mass isotopomer analysis for intracellular flux rates and various aspects of protein and amino acid metabolism. Specific applications (hyperinsulinemic euglycemic clamp) for use in humans and animals will be presented. The course uses a lecture format combined with problem discussions. In addition a popular aspect of the course is that trainees meet one on one with course faculty to discuss their specific research project and can present their project in an evening session. A new addition is detailed theory and experimental protocols to monitor specific pathways in an educational webinar style format that will be available on the course website. Typically, 70-80 trainees attended the course each year. Feedback from attendees has been very laudatory. A number of trainees have developed new research projects using isotope technologies. Thus, this popular course builds on and reinforces fundamental skills needed to study metabolic processes that are relevant to the mission of NIDDK.

NIH Research Projects · FY 2026 · 2024-03

The central thesis of the Vanderbilt AUD Research and Education Center (VAREC) is that effective treatment requires recognition of, and deep understanding of the distinct symptoms, etiologies, and disease progressions that underlie what is unitarily referred to as Alcohol Use Disorder (AUD). We propose a reverse translational “precision neuroscience” approach. We will dissect AUD endophenotypes to generate human circuit construct validated animal models that we will utilize to gain mechanistic insights and test therapeutic targets. VAREC will consist of 4 research components (Projects1-4) supported by Administrative and Research cores, as well as a Dissemination core. Project 1 (Blackford) expands our initial human imaging studies into a deep sampling approach to analyze BNST and network connectivity in AUD across the abstinence timespan, exploring individual differences in these networks with AUD relevant domains such as anxiety and depression. Project 2 (Winder) will build off an already active collaboration with Project 1 to perform reverse translational mouse studies to test emergent hypotheses on insula-BNST and hippocampal-BNST connectivity in mouse models of AUD, and to explore potential time-dependent therapeutic approaches during abstinence. Project 2 will also perform specific broad-scale mouse brain imaging studies to define network nodes for Project 1 to explore in years 4-5. Project 3 (Calipari) will explore whether alterations in negative affect observed in Project 1 and Project 2 lead directly to functional enhancement of negative reinforcement-oriented circuitries and behavior. Finally, Project 4 (Siciliano) will engage in deep phenotyping of individual differences in mouse behaviors correlated with compulsive drinking behavior, working across the projects to identify populations of precise behaviors that are predictive of compulsive ethanol seeking. Projects 2 and 4 explore therapeutic targets through analysis of endocannabinoid and dynorphin signaling. The project interactions will be coordinated by an Administrative core. A Research core will provide mice that have undergone one of two voluntary alcohol exposure models, Chronic Drinking-Forced Abstinence (CDFA) or Structured Tracking of Alcohol Reinforcement (STAR). It will also provide research infrastructure and computational support. Through a Dissemination core, VAREC will play an important role in public health by activities aimed at destigmatizing AUD treatment seeking and increasing prevention through novel nearpeer targeting of adolescents. This core will also allow VAREC to serve as a resource to the alcohol research community to facilitate mainstream incorporation of cutting-edge tools into the field.

NIH Research Projects · FY 2025 · 2024-03

Project summary Mounting evidence indicates that vascular damage and blood-brain barrier (BBB) dysfunction contribute to the progression of Alzheimer’s disease and related dementias (ADRDs). However, it remains challenging to study and understand connections between BBB dysfunction and ADRDs. In vitro BBB models represent valuable tools for investigating vascular contributions to ADRDs due to higher throughput and ease of manipulation. Historically, in vitro BBB models were constructed from primary brain microvascular endothelial cells (BMECs)— the principal functional component of the BBB—but after removal from the brain, BMECs rapidly de-differentiate and lose BBB-specific properties that are crucial for studying ADRDs. Strategies have been developed to differentiate human induced pluripotent stem cells (iPSCs) into endothelial cells with BBB attributes, but such approaches remain imperfect even after a decade of refinement. For example, iPSC-derived cells with a strong BBB phenotype lack robust endothelial character, but artificial reinforcement of endothelial identity via transcription factor overexpression completely ablates BBB properties. Conversely, efforts to imbue endothelial progenitors or mature endothelial cells with a BBB phenotype, through transgene overexpression or small molecule and growth factor treatments, have been largely ineffective. In vivo, BBB development is driven specifically by Wnt7a, and this Wnt ligand can only be transduced into active β-catenin signaling when membrane receptors GPR124 and RECK are present on the endothelium—for example, deletion of Wnt7a, GPR124, or RECK activity yields defects in BBB and neurovascular development. Given the importance of this pathway for BBB development, we hypothesize that specific activation of Wnt7 signaling in naïve iPSC-derived endothelial progenitors will impart representative BBB identity more effectively than prior efforts using generic activation of Wnt/β-catenin signaling. We will investigate this hypothesis using CRISPR and synthetic biology approaches to activate Wnt7 signaling in naïve iPSC-derived endothelial progenitors in simple two-dimensional models (Aim 1) and more complex three-dimensional neurovascular assemblies (Aim 2). Outcomes from this project are expected to yield improved in vitro human BBB models that are better suited for studying causes and impacts of vascular disturbance in ADRDs.

NIH Research Projects · FY 2026 · 2024-03

Project Summary/Abstract Type one diabetes (T1D) is an autoimmune condition characterized by the progressive destruction of insulin- producing β cells in the pancreas, leading to onset of clinical hyperglycemia and chronic dysglycemia. T1D can be managed for decades with constant blood glucose monitoring and administration of exogenous insulin. However, there is still no real cure to prevent or reverse onset of disease. Many current treatments have targeted the immune system to prevent action of autoreactive lymphocytes, however concerns for patients remain over immunosuppression. Additionally, recent literature has highlighted the level of β-cell dysfunction that also contributes to disease pathogenesis. Therefore, treatments designed to target both β-cell health and autoimmunity may provide the greatest potential for prevention and reversal of T1D. Prostaglandin E2 (PGE2), a lipid signaling molecule, possesses the capacity to modulate both the immune system and the β cells, through the action of two receptors, EP3 and EP4. These two receptors have near equal affinity for the PGE2 ligand, but couple to opposing intracellular pathways. Blockade of EP3 tone and enhancement of EP4 tone have previously been demonstrated to promote β-cell proliferation and survival in both mouse and human islets ex vivo. Additionally, in vivo, antagonism of EP3 enhances β-cell proliferation, mass, and oxidative stress responses in a mouse model of type 2 diabetes. In the pathogenesis of T1D, β cells are exposed to high levels of oxidative stress in the presence of proinflammatory cytokines from islet-infiltrating immune cells. This stress, along with direct destruction by cytotoxic T cells, results in a significant loss of β-cell mass. Therefore, modulating the EP3 and EP4 receptors pathways may be able to provide β cells significant protection against this attack to prevent loss or promote regeneration of β-cell mass. Furthermore, the primary function of PGE2 is to serve as an immunomodulator, and it can affect immune cell phenotype and function at various stages throughout the inflammatory response. At later stages of inflammation, sustained PGE2 production from macrophages helps facilitate resolution of inflammation, a process which is hypothesized to be disrupted in autoimmunity. Many of these pro-resolution activities are mediated via action of EP4, and therefore, enhancement of EP4 tone in the islet microenvironment may also promote β-cell survival by modulating immune cell activity and allowing proper resolution of inflammation. Ultimately, the proposed studies will elucidate whether targeting the opposing pathways of EP3 and EP4 can serve to mitigate β-cell death by promoting pathways associated with β-cell proliferation and survival, while simultaneously altering the islet microenvironment to favor a less proinflammatory state. This approach may unlock a novel therapeutic strategy for treating T1D by targeting both sides of the autoimmune equation at once.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY The objective of this application is to investigate the role of MeCP2 in the sustained antidepressant action of ketamine. Studies from our laboratory, as well as others, demonstrate that MeCP2 is a strong regulator of synaptic function with bidirectional changes in MeCP2 expression producing reciprocal alterations in neurotransmission. Studies have been examining how intracellular signaling mechanisms produce the acute antidepressant action of ketamine and transition to a sustained effect that may last over a week. This sustained ketamine effects requires the function of Methyl-CpG binding protein 2 (MeCP2), specifically MeCP2 Ser421 phosphorylation. We propose experiments to identify the mechanisms by which acute ketamine administration elicits MeCP2 phosphorylation at Ser421 and how this event is required for the sustained, but not the rapid effects of ketamine. A mechanistic understanding of the sustained effects of ketamine, and the underlying neurobiology, provide possible avenues to prolong antidepressant action and thus reduce the frequency of ketamine administration and potential adverse effects.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Age-related heart valve disease is the 3rd leading cause of cardiovascular disease and is especially prevalent among the elderly. Studies have shown that degenerative aortic valve disease affects over 25% of people over 65 years of age, and eventually leads to calcific aortic valve disease (CAVD). There is no effective medical therapy that modifies the progression of CAVD. CAVD is believed to be primarily driven by the valve interstitial cells (VICs). Briefly, when these normally quiescent VICs become ‘activated’ via unknown mechanisms, they become myofibroblasts and express smooth muscle α–actin and cadherin-11 (CDH11). We have generated robust evidence over the past decade to advance CDH11 as a prime candidate for CAVD therapy. Our hypothesis is that CDH11 is the mechanobiological driver of the majority of cases of CAVD and that targeting it can halt the progression and possibly reverse CAVD. Therefore, we propose three aims to advance targeting of CDH11 for treatment of CAVD. Aim 1: Clarify the cell-cell-specific mechanism of CDH11 engagement that leads to CAVD. Aim 2: Determine if targeting CDH11 can halt or reverse AS progression. Aim 3: Evaluate circulating CDH11 as a biomarker to identify interventional timing in AS patients. At the conclusion of these aims, we will have clarified the precise cell-cell interaction by which CDH11 drives CAVD, established if targeting CDH11 after the development of CAVD can halt or reverse the pathology, and determined if circulating CDH11 can be used as a biomarker in the progression of AS to guide a future clinical trial.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT The proposed short course is designed to expose researchers in learning and developmental sciences to principles and best practices in research data management and data sharing, and to provide scaffolded support to help researchers to engage in data sharing. The work has four specific aims. Aim 1: To develop a short course training researchers in children’s learning and development about data management and data sharing. Aim 2: To deliver a five-day summer intensive short course to early and established career scholars and enroll them in an online community of practice (Q/A sessions and webinars) for the remainder of the year. Aim 3. To disseminate the work developed via aim 1 by developing an open educational resource on data management and data sharing for scholars in the learning and developmental sciences. Aim 4: To evaluate the proposed project in terms of the knowledge, beliefs, and behaviors. The course will be developed and provided by core project faculty and consultants with years of expertise in research data management and data sharing. This team of investigators is uniquely positioned to train researchers in the areas of data management and data sharing because we include the primary investigators of the LDbase; an NIH funded data repository, and the only domain-specific data repository for learning and developmental science.

NIH Research Projects · FY 2024 · 2024-02

PROJECT SUMMARY Mitochondria serve as a central signaling hub for innate immune responses. Disruption of mitochondrial function is a hallmark for various infections and chronic inflammatory diseases. The overall objective of this proposal is to define the molecular mechanisms that regulate mitochondrial membrane homeostasis and determine how membrane disruption promotes inflammatory cell death. Mutations in leucine-rich repeat kinase 2 (LRRK2) are associated with disrupted mitochondrial integrity and increased reactive oxygen species. They are also associated with increased susceptibility to hormonal breast cancer, Crohn's disease, and mycobacterial infection, strongly suggesting a role in innate immune function. Recently, the Watson lab set out to investigate LRRK2's role in peripheral innate immunity, focusing on a gain of function mutation, Lrrk2G2019S. Mitochondrial stress conferred by the Lrrk2G2019S mutation increases demand on the electron transport chain, which leads to excessive ROS production. This increased ROS triggers a new type of cell death where a protein canonically associated with pyroptosis, gasdermin D (GSDMD), can associate with mitochondrial membranes and cause necroptotic cell death. Aim 1 of this proposal will identify the minimal domain within GSDMD that targets mitochondrial membranes, enabling a better understanding of the molecular mechanisms underlying GSDMD's newly described role in necroptosis. Aim 2 will investigate the contribution of various aspects of mitochondrial dysfunction to GSDMD mitochondrial targeting and necroptosis, providing new insights into connections between the disruption of mitochondrial homeostasis and GSDM relocalization. With the goal of understanding how mitochondria are impacted by genetic mutations and/or stress, Aim 3 will measure relocalization of the mitochondrial inner membrane phospholipid cardiolipin in WT and Lrrk2G2019S macrophages and catalog mitochondrial lipids in WT vs. Lrrk2G2019S macrophages. Defining the molecular mechanisms that drive inflammation in the face of specific mitochondrial mutations will help enable therapeutic interventions designed to correct specific aspects of mitochondrial dysfunction associated with a variety of inflammatory, infectious, cardiac, and neurological disorders.

NIH Research Projects · FY 2026 · 2024-02

Abstract The homeostatic gut microbiota provides essential functions to multiple aspects of human health, including modulating the interactions between host and enteric pathogens. Perturbations, such as intestinal inflammation, can shift the beneficial microbiota to an imbalanced state frequently referred to as dysbiosis, which instead exacerbates inflammatory disease outcomes in susceptible individuals. As such, microbial resilience is crucial to maintaining the structural and functional stability of the gut microbiome in the face of perturbations. Despite the central role in host health, mechanisms underlying microbiota resilience remain largely unexplored. During intestinal inflammation, the host immune system impedes invading pathogens through the sequestration of iron, among other micronutrients, in a process termed nutritional immunity. While decades of research have described how pathogens utilize small iron-chelating molecules termed siderophores to survive and thrive in the iron-starved inflamed gut, commensal survival strategies during nutritional immunity remain largely unknown. Our preliminary studies suggested that the prominent gut commensal Bacteroides thetaiotaomicron (B. theta) can capture iron from siderophores produced by enteric pathogens. Additionally, B. theta can prioritize its iron expenditure through the activation of a small RNA-mediated iron-sparing response, thereby conserving the limited iron for essential cellular processes. I hypothesize that B. theta couples xenosiderophore piracy and iron-sparing response through the action of small RNA to maintain resilience in the iron-limited inflamed gut. I will test this hypothesis using genetic, biochemical, and computational approaches in tandem with in vitro growth assays and murine models of infectious colitis. Experiments proposed in Aim 1 will determine the contribution of xenosiderophore piracy to B. theta fitness, using in vitro growth kinetics and mouse models of intestinal inflammation. Experiments in Aim 2 will define the mechanism, regulatory targets, and fitness contribution of iron-responsive B. theta sRNA during iron-limitation in vitro and in vivo. This proposed work is innovative because it presents a heretofore unexplored microbial factor in microbiome structure during intestinal inflammation. This proposed work is impactful because establishing a model for iron regulation in B. theta will provide insights into how interphylum iron metabolism and intracellular iron homeostasis may broadly contribute to gut microbiota resilience in the inflamed gut.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Pulmonary arterial hypertension (PAH) is a progressive lethal disease characterized by widespread obstruction in the smallest arteries of the lungs. Pulmonary microvascular obstruction leads to increased pulmonary vascular resistance, which subsequently causes right heart failure. Potential new targets for PAH therapy are lacking, yet despite the existing need for disease-modifying therapeutics, the rational selection of targets is significantly hampered by the still poor understanding of the cellular and molecular mechanisms that mediate PAH pathogenesis. We have shown in multiple animal models of PAH that targeting the serotonin 2B receptor (5- HT2B) genetically and pharmacologically prevents the development of disease. While 5-HT2B antagonism shows tremendous promise for PAH therapy, currently embodiments penetrate the blood-brain barrier. 5-HT2B mutation in the central nervous system is associated with several adverse consequences, such as depression, aggression, impaired sleep, and suicidality that limit the clinical potential of this treatment. Therefore, to overcome this limitation we have developed novel 5-HT2B antagonists that are both selective and potent, but also systemically restricted such that they do not cross the blood-brain-barrier. Here, we propose to de-risk these compounds for clinical translation through lead compound selection, safety and toxicity studies, and finally efficacy in preclinical models. At the conclusion of these aims, we hope to have identified a well-positioned, disease-modifying PAH therapy that has completed IND-enabling studies.

NIH Research Projects · FY 2026 · 2024-01

Project Summary Type 2 diabetes (T2D) is caused by the collapse of glucose homeostasis mechanisms, and the odds of devel- oping T2D increase during old age. Beta cells make insulin, a major hormone involved in the regulation of glucose homeostasis; these cells modulate insulin release in response to changes in blood glucose levels and/or meta- bolic demands. During T2D and aging, beta cell function and identity become compromised due to activation of stress pathways. Recent studies by us and others show that aging beta cells experience an underlying state of chronic endoplasmic reticulum (ER) stress associated with compromised autophagic flux and reduced expres- sion of beta cell identity markers, while reversal of ER stress rescues beta cell function and identity. It is currently unknown how beta cell heterogeneity is modulated during the adaptation process in response to changes in metabolic demands, and how these aspects are affected by aging and/or ER stress. In addition, the spatial organization of the beta cell epigenome, and its correlation with beta cell gene transcriptional heterogeneity, at the single cell level remains unknown. This project has the overarching goal of investigating how beta cell epi- genetic, transcriptional, functional, and cell longevity landscapes are affected by metabolic modulation intro- duced by changes in diet composition (e.g., calorie restriction (CR), high-fat diet), or during aging and/or ER stress. Moreover, we will investigate fundamental aspects regarding the spatial (in situ) organization of the beta cell (epi)genome in 3D, and how this aspect correlates with heterogeneous gene transcriptional patterns. The experiments in this proposal will leverage single cell multiome transcriptomics, high-resolution light, electron and stable-isotope microscopy, and quantification of in vivo beta cell function to determine the spatial, molecular, and functional landscapes of mouse and human beta cells exposed to different metabolic challenges. It is anticipated that CR will promote beta cell longevity and health by increasing the expression of beta cell identity while reduc- ing ER stress; this feature is expected to enable CR to mitigate the loss of beta cell identity and function observed in aging human beta cells. Understanding how beta cells adapt to changes in metabolic demands and/or ER stress to maintain their long-term function and health can lead to new ways to promote and/or preserve beta cell function in aging patients as well as in those living with T2D.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT The overall objective of this proposal is to elucidate the cellular mechanism and therapeutic potential of targeting the serotonin-2B receptor (5-HT2B) to attenuate fibrosis in hypertrophic cardiomyopathy (HCM). HCM is the most common monogenic heart disease and the most common cause of sudden death in young adults. A hallmark of this disease is myocardial fibrosis, an early manifestation of HCM defined as pathological remodeling that leads to heart wall stiffening and diastolic dysfunction. Due to the lack of knowledge on HCM disease emergence and progression, there is an unmet need to identify new pathways and therapeutic opportunities targeting fibrosis in HCM. Cardiac fibroblasts (CFs) are the primary cell type that contribute to fibrosis, as their function to preserve the myocytes’ surrounding environment by degrading and synthesizing the extracellular matrix can lead to pathological remodeling of the myocardium. Previous studies have antagonized 5-HT2B in cardiopulmonary diseases and demonstrated a decrease in fibrosis; in a mouse model of myocardial infarction, 5-HT2B antagonism decreased both the fibrotic scar size and number of isotropic collagen fibers in the myocardium, preventing the fibrotic remodeling process and preserving systolic and diastolic function. Our preliminary data shows an increase in 5-HT2B expression in HCM patients and a correlation between 5-HT2B and ANP, a marker of cardiac hypertrophy. In our hands, the 403/+ mouse model of familial HCM confirms an increase in 5-HT2B expression and pro-fibrotic markers. Therefore, we hypothesize that ablating 5-HT2B activity will attenuate fibrosis in HCM. To investigate this hypothesis, the following aims will be addressed. 1) Clarify the CFs vs. cardiomyocytes contribution to attenuation of fibrosis in HCM through global and CF-specific genetic ablation of 5-HT2B. This will be tested through the creation of two mouse models that provide both global and CF-specific genetic deletion of 5-HT2B. 2) Test the therapeutic efficacy of 5-HT2B antagonism and subsequent mechanical characterization of the cell type responsible for 5-HT2B – mediated fibrotic remodeling in HCM. Collaboration with Dr. Craig Lindsley has led to the discovery of two novel compounds that are highly specific for 5-HT2B and are systemically restricted from entering the brain. Following 5-HT2B antagonism, CFs will be isolated and mechanically characterized for their proliferative, contractile, and migratory capabilities. The training received throughout this fellowship will enable the candidate to conduct a successful project that will enhance our understanding of the underlying pathology and therapeutic potential of 5-HT2B antagonism in hypertrophic cardiomyopathy.