George Washington University

NIH Research Projects · FY 2025 · 2021-09

Glycans and glycoconjugates are critical to cellular communication, immunity, development, and disease processes, but remain underrepresented in biomedical data ecosystems. This project integrates GlyGen's glycoconjugate, glycan array, and protein variation data into the NIH Common Fund Data Ecosystem (CFDE), ensuring the unique data types are interoperable, findable, accessible, and reusable. The effort directly supports the CFDE mission to enable cross-domain biomedical research and advance precision medicine. The project introduces extensions to the data model for protein site-specific variation and glycan-protein datasets. The project will also prepare data in formats that support next-gen analytical methods, including machine learning and knowledge graph development, making the resources adaptable and broadly usable. The development of documentation, reusable workflows, and training materials will enhance reproducibility, interoperability, and community adoption. The GlyGen team will perform quarterly submissions of GlyGen metadata and datasets to the portal, with persistent access maintained through resolver services. Usage statistics and feedback will be collected to monitor adoption and guide improvements. Active engagement in steering committees, working groups, and public webinars will ensure alignment with consortium priorities. The project will participate in the Ontology Working Group and implement recommendations that will enable connecting GlyGen data with other DCC data and facilitate metadata submission. The team will provide expert suggestions on RFCs, contributing to consortium standards development. Education and training efforts will be coordinated with the Training Center to enhance workforce development and increase adoption of resources.  Sustained mechanisms for access, usage monitoring, and outreach will be established. By integrating GlyGen data into CFDE, researchers will gain access to detailed maps connecting glycans, genes, proteins, and other -omics data. The integration will support unique glycan use cases, including exploring sequence variants in glyco-related genes, understanding glycan epitopes, and investigating glycosyltransferases in rare diseases. Collaboration with CFDE-DCCs will facilitate data exchange and tool sharing, enabling multi-disciplinary queries and hypothesis testing that were previously impossible, now possible through CFDE.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Current techniques for measuring the physiological or psychological effects of obesity, and in particular bariatric surgery, either lack sensitivity, are invasive, or require expensive specialized devices. For measuring the physiological effects, BMI is commonly used to diagnose obesity despite the known shortcomings as a marker for metabolic syndrome. Biomarkers such as anthropometric measurements (e.g., waist-to-hip ratio) and visceral adipose tissue (VAT) have been shown to be superior to BMI in predicting health risks associated with obesity, but these measures lack sensitivity, specificity, or are expensive. Another key morbidity associated with obesity is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). The gold standard to the diagnosis and assessment of these conditions is histologic evaluation of liver biopsies. Fibroscan transient elastography is a noninvasive test which can also assess fibrosis, but high BMI and severe steatosis can decrease its accuracy. These approaches are either invasive, expensive, or relatively inaccurate. Measures of self-body perception are commonly used to assess psychological aspects of obesity, as body image is an important motivator for diet, physical activity, and weight loss intention. Body image perception is commonly assessed using self-reports and cartoon-like line drawings which are non-subject specific and insensitive. Based on our preliminary work (R21HL124443) that developed optical body scanning technology to capture 3D body shapes using inexpensive hardware, we propose to use the technology to study the physiological and psychological effects on subjects with severe obesity: · Develop prediction algorithm for hepatic steatosis, fibrosis, adiposity, and blood biomarkers using optical scans. The optical scan and biopsy data will be used as the training and validation set to develop a machine learning algorithm to cheaply and non-invasively predict the biomarkers from 3D body surface data. · Develop the use of optical scans for measuring the physiological effects of obesity. We will conduct a cross sectional and longitudinal study to further assess the use of optical surface scans to determine health indicators associated with obesity (using data from surface scan, DXA and serum biomarkers) for one year following surgery. We will establish a database of such data along with software for data mining the database. · Develop the use of optical scans for measuring the psychological effects of obesity. We will collect 3D surface geometry of each subject using optical scans and morph these to produce subject-specific images of larger or smaller body shape. We will use these images to study perception related to obesity and in particular patients undergoing post-operative body changes

NIH Research Projects · FY 2025 · 2021-09

HIV/AIDS once constituted a lethal diagnosis that ravaged many communities and has now become for some a chronic disease with associated co-morbidities. Likewise, our historic research successes in deciphering the mechanisms of fundamental HIV pathogenesis must now be directed to new questions about HIV elimination, vaccines and therapeutic development, and efforts to treat and prevent subsequent disorders among people surviving the disease. Researchers focused on these questions, with specific skillsets and knowledge, attractive to multiple career paths, are needed to address these emerging challenges in HIV/AIDS disease. The overall goal of this T32 is to prepare doctoral students with the knowledge, methodologic, analytic and leadership skills to become successful future HIV research investigators. To achieve these goals, we will (A) Provide intensive didactic education in HIV/AIDS research. A new required course in HIV Persistence will be offered to complement a core didactic research curriculum. Research in progress will be shared among trainees and experts at an annual retreat, and a catalog of elective courses will round out opportunities to build HIV knowledge in three major emphasis areas: (i) cure research, including T-cell therapy and the reversal of viral latency; (ii) co- morbidities, including malignancies and CNS disease; and (iii) prevention research, including vaccines and novel therapeutics; (B) Recruit two outstanding doctoral students per year for 2 years of support with activities to accelerate their progression to independence. Each trainee will have an individualized project characterized by a) multiple-mentor teams to aid in the design and implementation of hypothesis-driven research projects, b) a curriculum to build HIV specific knowledge and research skill sets, c) professional development including presentation skills and grant-writing d) strong exposure including participation in group-based externship opportunities in multiple career paths for HIV scientists, and strong programmatic oversight; and (C) Cultivate a local infrastructure and multi-departmental culture of collaboration to support HIV research training. This application joins faculty from multiple GW departments and schools, as well as the District of Columbia Center for AIDS Research (DC CFAR) to introduce new perspectives and fresh ideas that will collectively offer outstanding collaborative training in cross-cutting high priority areas of HIV research. Enhanced cross-school and regional investigator orientation and networks, and research progress reports and meetings are intrinsic to this plan. New approaches to enhance mentor development will be expanded and institutionalized to strengthen faculty mentoring practices in effective autonomy-supportive mentoring.

NIH Research Projects · FY 2025 · 2021-08

Summary Despite the development of potent anti-retroviral therapy (ART) that successfully suppresses virus replication in the majority of people living with HIV (PLWH), there is no treatment that can cure this infection entirely. The major obstacle in eradicating HIV is the persistence of various anatomical viral reservoirs (VRs), including the central nervous system (CNS), that have the capacity to produce infectious virus and systemically spread within a short period upon cessation of ART in all, with few exceptional cases. Therefore, developing novel interventions aimed at reducing or eliminating the VRs is one of the key priorities for HIV research. In response to RFA-MH- 20-701, our application proposes basic science and preclinical research in SIV-infected rhesus macaques (RMs) to model aspects of VR in the CNS-resident myeloid cells of PLWH, and to investigate the efficacy of the novel pharmacologic strategy to prevent establishment of HIV persistence in the CNS. Thus, based on the observations outlined in this application we hypothesize that the disruption of PMC formation during acute phase of infection will limit the seeding and maintenance of VR and, as a consequence, the extent of viral rebound in the CNS following analytical therapy interruption (ATI). Three aims are proposed: (1) To investigate whether the systemic disruption of PMC formation during acute phase of infection, regulates viral persistence in the CNS; (2) To investigate whether the systemic disruption of PMC formation during acute phase of infection, regulates the kinetics and extent of viral rebound after ATI; and (3) To investigate whether the systemic disruption of PMC formation during acute phase of infection, regulates the neuroinflammation and synaptodendritic damages associated with long-term ART and ATI. These aims will be achieved by (i) using a well-established model of SIV-infected RMs treated with suppressive ART, and (ii) performing in vivo Ab-mediated disruption of PMC formation during acute phase of untreated infection. Revealing the mechanisms through which platelets regulate the persistence of HIV in myeloid cells will provide a critical understanding of how these cellular interactions function in mammalian cells, and an insight into how a potential HIV cure can be achieved in PLWH.

NIH Research Projects · FY 2025 · 2021-07

The objective of this proposed research is to develop a noninvasive, challenge-free, and widely available method for quantitative mapping of cerebral oxygen extraction fraction (OEF). As the brain continuously consumes 20% of the total oxygen supply, oxygen deficiency easily causes severe brain tissue damages as in hypoxia in ischemia in stroke and Alzheimer's disease. Regional OEF is an essential, direct biomarker for tissue viability and function and highly desired for evaluating and stratifying treatments in these neurologic disorders. Widely distributed MRI provides the potential to overcome the 15O PET, the current reference standard but clinically not used due to its limited availability. In MRI, quantitative mapping of OEF requires estimating deoxyheme concentration [dH] from the MRI signal. Three major approaches have been proposed to estimate [dH] from MRI magnitude signal. However, they commonly suffer from poor sensitives and burdensome data acquisition schemes as the MRI magnitude signal which they utilize has a complex dependence on [dH]. Consequently, no MRI-based OEF mapping has been routinely used in clinical setting. Furthermore, none of these methods have been validated against the current reference standard, 15O PET. Recently, we developed a promising, novel, non-invasive MRI-based OEF method that requires no vascular challenge and utilizes a single routine MR sequence. By integrating quantitative susceptibility mapping (QSM) modeling of often neglected MRI phase signal and quantitative blood oxygenation level dependent (qBOLD) modeling of MRI magnitude signal, our model (QSM+qBOLD=QQ) can distinguish deoxyheme iron in venous vasculature from diffusive other susceptibility sources. In our preliminary data, QQ has been validated against 15O-PET in healthy adults and showed OEF abnormalities in ischemic stroke, multiple sclerosis, and brain tumor. However, for clinical use, data acquisition and processing scheme of QQ should be improved to ensure the accuracy of OEF as a lack of data at short echo time and a non-optimal signal modeling with gradient-based solvers in current setting hinders the accurate OEF estimation. In this K99/R00 project, we will establish a clinically readily applicable MRI toolset for quantitative OEF mapping, which is validated and available to every MRI scanner, by improving QQ. We will achieve this through 4 specific aims. Aim1. Develop optimal data acquisition for MRI-OEF mapping. Aim 2. Develop data processing algorithms for robust OEF estimation. Aim 3. Perform technical validation of MRI-OEF against 15O PET. Aim 4. Perform clinical validation of MRI-OEF in patients with intracranial stenosis. Our experience and preliminary data give us confidence that we will very likely succeed this this proposed project. In a timely fashion, the project will lead to a novel, validated, non-invasive, challenge-free, routinely usable and quantitative MRI OEF mapping, offering the potential to replace invasive, complicated current standard 15O PET OEF. This tool will lead to better understanding and management of neurovascular disorders, e.g. stroke.

NIH Research Projects · FY 2025 · 2021-07

Abstract Effective treatment of HIV infection has reduced the severity of HIV-associated neurocognitive disorder (HAND), however, the incidence of CNS neurological dysfunction (~50% of HIV patients) has not been diminished by the treatment. With dramatically extended life expectancy of HIV-infected patients, neurological dysfunction reduces the quality of life by affecting learning and executive functions, and puts these individuals at risk of developing significant health problem. The treatment options for this co-morbidity are limited by poor understanding of its pathogenic mechanisms in virologically suppressed patients. Several hypotheses have been suggested, ranging from low grade chronic neuroinflammation caused by HIV infection, to neurotoxicity of HIV-related factors, to HIV accelerating the natural development of known neurodegenerative diseases, such as Alzheimer’s disease. Although these hypotheses are consistent with some elements of HAND, none of them explains the full pathological manifestation of this disorder and its unique relationship with HIV infection. In this application, we propose to test a novel hypothesis that, if confirmed, will point to the key element of pathogenesis of CNS neurological disorder caused by HIV infection and may translate to novel treatment opportunities. We hypothesize that the central mechanism in HIV-associated CNS disorder is the reorganization of lipid rafts caused by HIV Nef. Changes in neuronal lipid rafts promote protein misfolding/aggregation, exacerbate inflammatory responses, and affect neuronal communications leading to functional impairment and eventually to neurodegeneration. Dysfunction of the lipid rafts is essential for pathogenesis of many neurodegenerative diseases, including Alzheimer’s, pointing to a broad relevance of our hypothesis to diseases of aging population. This hypothesis is based on our published and preliminary findings that HIV protein Nef reorganizes lipid rafts in macrophages and neurons. We recently demonstrated that changes to lipid rafts inflicted by Nef are similar to those found in neurons infected by prions. Importantly, recent studies have shown that neurons exposed to Nef-containing exosomes, released by HIV-infected brain macrophages, microglia and astrocytes, take up exogenous Nef, which is functionally active. Nef production in viral reservoirs, including brain, continues in the presence of antiretroviral therapy. The following aims will be pursued to test this innovative hypothesis. Aim 1: To establish the contribution of Nef to HIV-associated CNS neurological dysfunction in mouse models; Aim 2: To determine mechanisms by which Nef released from HIV-infected cells affects cholesterol metabolism in neurons, causing neurological dysfunction; Aim 3: To target lipid rafts as a potential therapeutic approach to treat HIV-associated neurological dysfunction. These interconnected but independent aims will provide an actionable model of HIV-associated CNS disorder.

NIH Research Projects · FY 2025 · 2021-07

In the U.S., training staff at community- based organizations (CBOs) to deliver psychological support has been highlighted as a way to increase availability of mental health services and increase access to mental health care. We propose to employ a mental health task-sharing model by partnering with CBOs in NYC to train CBO staff without professional mental health training to deliver mental health services. We will examine the impact of CBO staff delivering mental health services to improve mental health outcomes including reduced general psychological distress, depression, and anxiety, as well as improve economic outcomes including saving on healthcare expenditures. We propose a cluster randomized control trial (cRCT) in NYC comparing Services as Usual (SAU arm) and delivery of mental health services with Problem Management Plus (PM+) (Intervention arm) among CBOs offering services for housing, job training, financial literacy, and other social services promoting independent productive livelihoods. The target conditions will be psychological distress, depression and anxiety. We will evaluate mental health outcomes of populations served by CBOs integrating PM+ into their other activities compared to CBOs delivering SAU. We will evaluate implementation processes to inform policy recommendations for community-based delivery of psychological interventions and inclusion of mental health to improve outcomes of other social services (e.g., integration of PM+ to improve financial, employment, and other independent productive livelihood skills). Successful completion of these aims will contribute to the NIMH Strategic Plan. Specifically, a) employing implementation science to maximize the impact of research for improving effectiveness and reach of mental health services b) strengthening research-practice partnerships to expedite adoption, sustained implementation, and continuous improvement of evidence-based mental health services; and c) developing innovative service delivery models to dramatically improve outcomes of mental health services for the goals of promoting employment, housing stability, and promoting savings on healthcare costs.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract This UG3/UH3 proposal is in response to RFA-TR-20-031-Basket Clinical Trials of Drugs targeting Shared Molecular Etiologies in Multiple Rare Diseases. The proposed studies focus on two ultra-rare maternally inherited mitochondrial diseases MELAS (mitochondrial encephalopathy, lactic acidosis, stroke-like episodes) and LHON-Plus (Leber’s hereditary optic neuropathy-Plus). Both diseases are among those studied by the Rare Diseases Clinical Research Network. Patients do not have access to effective therapeutic intervention, resulting in significant disability, morbidity, and premature death. The devastation wrought by these diseases underscores the urgency to address this unmet medical need and develop novel therapeutic candidates. However, their ultra-rare prevalence makes it challenging to recruit an accrued number of MELAS and LHON- Plus patients to clinical trials. Thus, the proposed basket clinical trial design will combine these two ultra-rare populations to provide proof-of-concept of its feasibility for divergent patient populations. MELAS and LHON-Plus patients exhibit divergent and overlapping clinical neurological and non-neurological symptoms. They are caused by a maternally inherited pathogenic variant that results in a defective oxidative phosphorylation pathway responsible for mitochondrial ATP synthesis. Both diseases share the molecular etiology of Complex I deficiency, causing ATP deficiency and chronic energy deficit. We designed a novel two-pronged pharmaco-epigenomic strategy to increase ATP output in MELAS and LHON-Plus patients. Our pre-clinical studies using ex-vivo patient-derived fibroblasts demonstrate the feasibility of our lead compound to promote mitochondrial recovery in MELAS and LHON-Plus patient’s fibroblasts. The proposed multi-PI studies combines the cross-disciplinary strengths of the George Washington University School of Medicine and Health Sciences and Children’s National Medical Center, a referring site for the North American Mitochondrial Disease Consortium. This partnership is funded by an NIH Clinical and Translational Science Award UL1 Program providing a robust infrastructure for the proposed studies. Aim 1 (UG3 phase) focuses on translational MELAS and LHON-Plus studies and submission of an IND protocol to the FDA. Aim 2 (UH3 phase)focuses on a basket clinical trial with MELAS and LHON-Plus to: 1) provide proof-of-concept that the basket design can be applied to divergent ultra-rare diseases; 2) advance the dataset for safety and pharmacokinetics/pharmacodynamics of our lead compound for a larger number of patients than in a conventional clinical trial setting; and 3) gather outcomes and practical information for optimizing the design of future basket clinical trial. Our innovative design lies in applying the concept of basket clinical trial not only to multiple diseases with a common molecular target, but also to groups with similar ex-vivo fibroblasts response to butyrate across these diseases to improve our ability to evaluate our therapeutic drug in ultra-rare disease populations.

NIH Research Projects · FY 2025 · 2021-04

Project Summary Ovarian cancer (OC) is the fifth leading cause of cancer-related death among women and the deadliest gynecological cancer in the United States. The current standard treatment for ovarian cancer consists of surgery followed by platinum drug-based chemotherapy. Platinum drugs could induce DNA interstrand crosslink (ICL), which results in replication fork stalling and thus decrease cell viability. Although most of patients initially respond to platinun drug-based chemotherapy and achieve remission, up to 80% of patients become refractory to platinum drugs over time and ultimately succumb to the disease due to the resistance to platinum-based therapy. Thus, it is urgent to develop novel approaches to overcome platinum drug resistance of ovarian cancer (PROC). Fanconi anemia (FA) pathway is critical to repair ICLs and elevated activity of FA signaling is one of major mechanisms leading to platinum-resistance in ovarian cancer. The role of FA pathway in repair of ICL has been well studied last decade, however, how the regulatory switch from a stalled replication fork caused by platinum drugs to initiation of FA signaling and how FA is activated in PROC cells are poorly understood. To elucidate the mechanism regulating initiation of FA signaling, we have conducted the significant amount of preliminary studies to demonstrate that And-1 is critical for activation of FA signaling and FA- mediated platinum drug resistance in ovarian cancer. The major objective of this proposal is to determine the mechanism of how And-1-FANCM axis regulates FA signaling and platinum resistance in PROC. Here, we propose three specific aims. Aim 1: Determine how And-1 promotes the switch from stalled replication forks to initiation of FA signaling at ICLs. Aim 2: Determine the role of And-1 in the regulation of platinum drug resistance in PROC cells. Aim 3: Evaluate the effects of And-1 inhibition on platinum drug resistance using orthotopic PROC PDX and syngeneic models. The completion of proposed studies will not only elucidate a novel mechanism regulating platinum resistance in PROC, but also provide an innovative therapeutic strategy as well as a new potential drug for treatment of PROC patients.

NIH Research Projects · FY 2026 · 2021-04

Despite focused research efforts, the five year survival for ovarian cancer (OC) has remained unchanged for decades and novel therapies are urgently needed for this deadly disease. Therapies that activate the immune system to kill cancer cells, including anti-PD-1 checkpoint blockade therapy, have shown vigorous and durable responses, but the majority of patients, including those with OC, fail to respond. The underlying mechanism remains unclear. Repetitive elements (REs) comprise the majority (45%) of the human genome. In most somatic tissues, REs are silenced by DNA methylation and other epigenetic modifications to prevent their transcription. We demonstrated that treating OC cells with DNA methylation inhibitors (DNMTis) and histone deacetylase inhibitors (HDACis) increases immune signaling from tumors through demethylation of REs and production of RE double-stranded RNA to activate the interferon response. This signaling recruits CD8+ T cells to sensitize tumors to anti-PD-1 immunotherapy. REs translate proteins that can be targeted as tumor-associated antigens. Thus RE activation both promotes interferon signaling to reverse the immune- suppressive tumor microenvironment and presents potential tumor-specific antigens as T cell targets. The premise of this proposal is that P53 and epigenetic mechanisms regulate REs in cancer and thus mutant TP53 will affect immune signaling and response to epigenetic and immune therapy. Approximately half of all cancers have mutations in TP53, the gene encoding the P53 protein, 90% of which are “hotspot” mutations located in the DNA binding domain. These missense mutations encode functional proteins with reduced transcriptional activity at canonical cell cycle target genes that may also exhibit oncogenic gain of function transcriptional activity at new targets. High grade serous OC makes up about 70% of all cases and is characterized by nearly 100% mutant TP53. While the critical role of P53 in cell cycle regulation and apoptosis is known, P53 regulation of REs in cancer remains poorly defined. Approximately 30% of P53 binding sites are found in REs and our preliminary data show that P53 binds directly to REs. Further, we show that P53 hotspot mutant cell lines treated with DNMTi/HDACi exhibit significantly increased chromatin accessibility at REs and transcription of REs compared to TP53 wild type cell lines. We hypothesize that mutant P53 aberrantly activates REs, amplifying the RE-induced immune response. We will test this hypothesis via the following aims: In Aim 1, we will determine how wild type and mutant TP53 regulate REs to affect the DNMTi/HDACi-induced interferon response. In Aim 2, we will determine how p53 status affects the DNMTi/HDACi-induced T cell response and sensitization to immune therapy in a mouse model of OC and a clinical trial of OC patients treated with immunotherapy. In Aim 3, we will evaluate REs as tumor antigens in different P53 backgrounds. Results of this innovative work will answer novel basic science questions about REs and P53 and open new directions for epigenetic and immune therapy in OC.

NIH Research Projects · FY 2025 · 2021-04

ABSTRACT In the US, an estimated 3.3 million children experience dizziness and balance problems (19). Children with congenital vestibular disorders (CVDs) show delayed motor development and challenges in maintaining posture and balance, indicating that the vestibular neural circuitry is affected. Computed tomography (CT) shows that children with CVDs most commonly form a sac-like inner ear with the semicircular canals missing or truncated. It is not known how their vestibular connectivity is altered. We hypothesize that formation of a sac-like inner ear during early gestation results in a reduced number of vestibular ganglion cells forming fewer primary vestibular synapses on hair cells peripherally and on vestibular nuclei neurons centrally, leading to underconnectivity in the vestibular system. We further hypothesize that the sac-like inner ear pathology results in abnormal convergence of canal and otolith fibers onto vestibular nuclei neurons, or anomalous connectivity, contributing to abnormal signal processing in these neurons. The proposed work will establish a framework to test the overarching hypothesis that formation of a congenitally-malformed, sac-like inner ear alters the peripheral and central vestibular neural circuitry. To address these questions, our laboratory has implemented and validated a new chick embryo model. We can produce a reproducible animal model in 85% of cases by surgically rotating the developing inner ear or “otocyst” 180° on one side in two-day old chick embryos (E2). Since the procedure involves Anterior-posterior axis Rotation of the Otocyst to produce a Sac-like inner ear, the model is called the ARO/s chick. The sac-like inner ear of ARO/s chicks resembles the sac-like inner ear in children with CVDs. After hatching (H), ARO/s chicks experience challenges in maintaining balance and walking. As a first step in understanding the consequences of the sac-like inner ear on the developing vestibular neural circuitry, in Specific Aim 1 we will further analyze the vestibular epithelium and quantify vestibular ganglion cells to determine to what extent primary vestibular afferent synapses are decreased in ARO/s chicks. In Specific Aim 2, we will combine imaging and electrophysiological approaches to determine whether a structurally-uniform class of vestibular nuclei neurons, the principal cells of the tangential nucleus (TN), acquire the orderly inputs from canal and otolith fibers, passive and active membrane properties, and synaptic transmission found in normal chicks. In Specific Aim 3, we will perform ethological tests to characterize posture and balance in H5 ARO/s chicks, followed by the horizontal vestibuloocular reflex (hVOR) using Earth vertical axis rotation (EVAR) to test canal function and the static tilting platform test to evaluate otolith function. The experimental outcomes will provide a foundation to better understand the pathological changes occurring in the vestibular neural circuitry of CVD patients, and modify our thinking on how to treat these disorders.

NIH Research Projects · FY 2025 · 2021-03

Project Summary This resubmission proposal aims to elucidate the role of a histone deacetylase, HDAC11, in diseases such as multiple sclerosis (MS), and to establish HDAC11 inhibition as a potentially effective new treatment strategy for diseases including MS. MS is a chronic, immune-mediated demyelinating disease of the central nervous system. Like many autoimmune disorders, it presently has no known cure, and current drugs available for managing this disease are only effective early on and are accompanied by many adverse effects. The disease mechanism of MS remains unclear, and no effective targeted therapy is available for chronic progressive MS. Our preliminary studies show that deletion of HDAC11 ameliorates clinical symptoms in a mouse model of MS. In parallel, we discovered a novel HDAC11 enzymatic activity that is >10,000-fold more efficient than its deacetylase activity. This novel activity allows us to begin to uncover physiologic substrates of HDAC11, which in turn will help to uncover the biological mechanisms of HDAC11’s actions. One of the goals of this research is to investigate how this newly discovered enzymatic activity underlies the immune-regulatory function of HDAC11 in MS. Knowledge gained from these studies will help to further understand the disease mechanism of MS and to develop better therapeutics. Because the discovery of a novel HDAC11 activity has enabled us to develop, for the first time, HDAC11-specific inhibitors, the chief objective is to further improve these inhibitors and test whether they can be used to treat diseases such as MS in our established mouse models. Our multidisciplinary team has expertise in all aspects needed to make this project successful. Overall, the proposed studies in this application will not only yield a better understanding of HDAC11’s function in health and diseases, but may also result in a first prototype targeted therapy for the treatment of chronic progressive MS, and possibly other diseases as well.

NIH Research Projects · FY 2026 · 2020-12

Project Summary The District of Columbia Clinical Trials Unit (DC CTU) will conduct clinical trials for persons with and at risk for HIV in DC, where 1.8% of the overall population and 2.7% of the African-American population are living with HIV. The DC CTU will unite two existing, high-performing DAIDS/NIAID Clinical Research Sites (CRS), the Whitman-Walker Health CRS and the George Washington University CRS, in a new CTU, participating in the HIV/AIDS Adult Therapeutics and HIV Prevention Clinical Trials Networks. Our innovative and geographically- focused DC CTU will ensure exemplary clinical trials management, recruitment, and retention of a diverse sample of participants, with a community engagement strategy built on existing partnerships and Good Participatory Practices. The DC CTU will achieve four specific aims: 1) Provide scientific leadership and administrative infrastructure to allow high-quality conduct of therapeutic and prevention studies at two established CRSs; 2) Implement best practices to ensure continued high recruitment and retention rates; participant safety; and laboratory, pharmacy, data, and regulatory excellence; 3) Actively engage with the local community to ensure optimal enrollment of diverse participants from within and beyond clinic settings using Good Participatory Practices; and 4) Capitalize on the resources available from the DC CFAR and DC Cohort to engage senior investigators, mentor investigators in clinical trials conduct, and access a broad array of services to address NIH and network scientific and programmatic priorities. The DC CTU will engage participants who can benefit from innovative clinical trials while we contribute to high-quality participatory research that will eventually end the HIV epidemic.

NIH Research Projects · FY 2024 · 2020-07

In order to achieve the end of the HIV epidemic, concerted efforts will be needed to address the HIV care continuum, including improving retention in care and viral suppression among persons living with HIV (PLWH). Currently, in the U.S., it is estimated that less than 50% of PLWH are retained in care and even fewer are virally suppressed. Studies have shown that these PLWH have poorer clinical outcomes and are at risk of transmitting HIV to others, hence the need for innovative solutions to improve retention in care and subsequent viral suppression. Theory-based mHealth interventions have been shown to be promising in reaching these at- risk groups and improving HIV-related outcomes. PositiveLinks is a clinic-deployed mHealth platform that includes patient and provider smartphone apps, a web portal for clinic staff and providers to manage patient cohorts, an online implementation guide, and a learning management system to train and certify clinic staff. It has theory-based features including daily queries of adherence, mood, and stress, graphical feedback for self- monitoring, secure messaging with staff, appointment reminders, anonymized peer support, information resources, and document upload capability to support insurance re-enrollment. A 12-month prospective study in poorly retained adults with HIV found that PL increased RIC and VS, with app usage related to benefit as well as improved social support and stigma. PL is a promising existing mHealth tool for PLWH, but its efficacy has not been tested in a rigorous randomized trial, nor in urban populations. We propose to test the efficacy of PositiveLinks to improve retention in care and viral suppression among a cohort of PLWH in a high HIV prevalence city of Washington, DC. Participants will be identified from the DC Cohort, a longitudinal observational prospective cohort of PLWH receiving HIV care at 14 clinical sites in Washington, DC. First, we will conduct formative research to assess the feasibility, acceptability and usability of PositiveLinks among this urban cohort and conduct subsequent adaptations based on these findings. We will then conduct an efficacy study through a cluster randomized controlled trial at 14 DC Cohort sites among 560 PLWH. Clinics will be randomized to PL or usual care. Our primary outcomes will include viral suppression, retention in care and visit constancy at 12 months. Finally, we will conduct mixed methods implementation science research guided by the Consolidated Framework for Implementation Research and RE-AIM to identify site, patient, provider, and system factors that characterize best practices in program implementation. If successful, this research will lead to the development of a novel and efficacious approach to improving retention in care and viral suppression among PLWH which could lead to next-generation dissemination research that will contribute to HIV epidemic control. This project is responsive to NIH priorities, National HIV/AIDS Strategy, and Ending the HIV Epidemic goals as it is cross-cutting, seeks to reduce health inequities, and to improve health outcomes to achieve sustained viral suppression in a geographic hotspot for HIV.

NIH Research Projects · FY 2024 · 2020-07

PROJECT SUMMARY/ ABSTRACT The overall goal of the proposed George Washington-Cancer Biology Training Program (GW- CBTP) is to educate and train scientists who will advance research on the molecular and genomic causes, progression, treatment and relapse of human malignancies. This application builds upon the formation of the George Washington Cancer Center (GWCC), which supports a vibrant cancer research community of over 150 members with a vision to create a cancer-free world through groundbreaking and rigorous research, innovative education and equitable care for all. As the only cancer training program at GW, the CBTP will reflect the major cancer research strengths of the GWCC that include Cancer Immunology and Immunotherapy, Cancer Signaling & Genomics and Cancer Epigenetics & Technology. This application seeks to appoint 3 postdoctoral and 1 predoctoral trainee annually and provide each with an individualized two year program of three elements: a discovery research project guided by expert cancer researchers and clinician investigators; a curriculum of courses and workshops on state-of-the- art approaches and emerging therapies; and career and professional development, including experience in grant writing and presentations. Key to this program is an institutional commitment to address the prominent cancer health disparities in breast, cervical, colorectal, pancreatic, liver and prostate cancers in the District of Columbia. Outstanding faculty preceptors who are all leaders in their fields and maintain outstanding track records of research productivity and mentoring of young scientists will serve as research mentors in a dual-mentor model with clinician scientists. Postdoctoral fellow appointees will have completed an M.D. or Ph.D. degree and have a clear commitment to pursuing independent careers in cancer research. Predoctoral students in their second year of graduate study in the doctoral program in Cancer Biology will be eligible for appointment. All trainees will have demonstrated a significant interest in pursuing a career in cancer-related research. The program will provide highly sought-after training experiences to transform promising young researchers into future independent cancer research scientists.

NIH Research Projects · FY 2025 · 2020-06

Abstract Latent toxoplasmosis continues to be a problem for immunocompromised infected individuals and toxoplasmic encephalitis (TE) is one of the most common life threatening central nervous system infections in these patients. The reactivation of latent toxoplasmosis is attributed to lack of adequate CD4 T cell help that compromises the CD8 T cell immunity against the parasite. Similar to humans, mouse models of toxoplasmosis have demonstrated the critical role of CD4 T cells for the maintenance of robust CD8 T cell immunity. In a recent study we demonstrated that CD8 T dysfunction leading to reactivation in chronically infected host is a consequence of CD4 T cell exhaustion. Treatment of chronic host with antigen-specific non-exhausted CD4 T cells can restore CD8 T cell functionality and prevent reactivation. Interestingly, CD4 exhaustion is linked to up-regulation of transcription factor BLIMP-1, which causes an increase in the expression of inhibitory receptors on these cells. Preliminary data for the proposal suggests that during latent toxoplasmosis increased BLIMP-1 expression leads to epigenetic changes in antigen-specific, CD4 TCM (central memory) subset. The transcription factor gains accessibility to chromatin sites on this population and changes their epigenetic landscape. BLIMP-1 ablation re-invigorates CD4 T cells due to downregulation of inhibitory receptors and increased expression of positive co-stimulatory molecules. The proposal has two specific aims. In aim 1 we will determine the chromatin accessible sites on CD4 TCM that BLIMP-1 binds to. We plan to define the epigenetic changes in CD4 TCM during latent toxoplasmosis that leads to their exhaustion. In aim 2 studies will be performed to evaluate if restoration of CD4 T cell function due to BLIMP-1 ablation is dependent on the up-regulation of 4-1BB and OX40 or other costimulatory molecules identified in aim 1. We will determine if cell intrinsic signaling by these co- stimulatory molecules is required for optimal recovery of CD4 T cell function in BLIMP-1 ablated cells. Finally, studies will be performed to determine if CD4 T cells treated with agonist for co-stimulatory molecules downregulate inhibitory receptors on CD8 population and confer strong effector cytotoxic program on these cells that is critical for containing chronic toxoplasma infection.

NIH Research Projects · FY 2024 · 2020-06

Project Summary/Abstract Recent developments in the field of immunotherapy clearly support the contribution of the immune system in eradicating cancer. Histone deacetylase (HDAC) inhibitors are currently employed in the treatment of many malignancies, and accumulating evidence suggests that many of the anticancer effects of HDAC inhibitors involve the immune system. Previously, there was limited information on the role of HDAC11 in immunity and cancer. We discovered that HDAC11 negatively regulates IL-10 production in antigen-presenting cells. We also found that HDAC11 is highly expressed in T lymphocytes and neutrophils and, subsequently, revealed that HDAC11 disruption in T cells is associated with an enhanced pro-inflammatory cytokine profile and effector molecule production. T cells lacking HDAC11 are less susceptible to regulatory T cell suppression in vitro, are refractory to tolerance induction in vivo, and display enhanced anti-tumor responses in transplanted mantle cell lymphoma murine models. Furthermore, HDAC11 is a multifaceted regulator of neutrophils. The absence of Hdac11 in neutrophils significantly increases cellular production of proinflammatory cytokines and promotes cell migration and phagocytic capacity. More recently, our group discovered an efficient novel activity for HDAC11, the removal of long-chain fatty acyl groups from protein lysine residues. This novel activity is >10,000-fold more efficient than its deacetylase activity. Using a syngeneic mouse-to-mouse model, we established ectopic tumors in Hdac11 wildtype and knockout (KO) mice. The growth of the syngeneic lymphoma cells in the Hdac11 KO mice was markedly inhibited, pointing toward a crucial role of HDAC11 in the tumor microenvironment. In this resubmission application, the central hypothesis is that HDAC11 reprograms anti-cancer immunity via its defatty-acylation activity and presents a potential novel drug target for cancer treatment. Our long-term goal is to develop a detailed molecular understanding of HDAC11's role in anti-tumor immunity. Results from this work will: (1) provide a better understanding of the anti-tumor behavior of HDAC11; (2) expand a functional understanding of protein lysine defatty-acylation in cancer; (3) develop better treatment strategies for cancer through targeting the lysine defatty-acylation mechanism; and (4) produce selective HDAC11 inhibitors, which will accelerate the development of new cancer treatment strategies.

NIH Research Projects · FY 2025 · 2020-06

Washington, DC is an HIV “hotspot,” as it is one of the 50 jurisdictions in the US that accounts for more than 50% of new diagnoses. With 11,904 people with HIV (PWH) residing in DC, the epidemic disproportionately impacts certain populations and reflects an aging epidemic. As we strive to end the HIV epidemic, it is critically important to characterize persons who are at risk for HIV, monitor the use of novel prevention tools, identify persons newly diagnosed with HIV, and effectively direct HIV treatment efforts. The DC Cohort, a longitudinal cohort study of over 12,800 PWH receiving care at 14 clinical sites in DC, is uniquely positioned to help end the HIV epidemic. Launched in 2011 with the goal of bending the trajectory of the HIV epidemic in DC by improving the quality of care, the Cohort was established as an R24 research platform in 2019. The Cohort engages academic and community-based clinics, links to DC Health Department databases, and provides near real-time performance feedback to clinics on HIV outcomes using a data visualization Clinical Dashboard. All of this is achieved through automated data extraction and harmonization of electronic medical records data from varied clinical settings and across age groups. In this renewal application, we aim to enhance our existing population-level database to improve the longitudinal characterization of care and outcomes using a status neutral approach by enrolling both people at risk for and living with HIV. We will continue our DC Health data linkages to improve our ability to describe and monitor HIV and co-morbidities outcomes to help inform local and national efforts to end the HIV epidemic. Our second aim is to use cutting edge research to identify factors and strategies to monitor and improve HIV prevention for people at risk for HIV and care and treatment for those living with HIV in DC and beyond. By continuing to triangulate data from the core database, patient reported outcomes surveys, and our various data linkages, we will describe status neutral care outcomes aligned with local and national research priorities. New data on those at risk will be used to monitor and describe pre-exposure prophylaxis use, demographics, and persistence as well as HIV incidence. We will expand the clinical Dashboard to monitor key programmatic and clinical indicators and identify strategies to improve the quality of care. Finally, we will leverage the DC Cohort as a valuable and dynamic population- based resource to support the development and testing of pioneering interventions to optimize HIV prevention, clinical care, and treatment. The goals of the Cohort align with OAR and NIAID priorities that focus on reducing HIV incidence, achieving viral suppression among PWH, and improving the management of HIV-related clinical outcomes. The DC Cohort is an unparalleled clinical, research, and public health resource that enables us to monitor care in a high prevalence city. Lessons learned will produce generalizable knowledge that can be applied in other urban hotspots across the US.

NIH Research Projects · FY 2025 · 2020-01

PROJECT SUMMARY/ABSTRACT Advances in genetics, molecular biology, and cognitive neuroscience offer promise towards personalized treatment and improved outcomes in individuals with Autism Spectrum Disorder (ASD). However, the promise of precision medicine has been hindered by a lack of mechanistic models that explain phenotypic and etiological heterogeneity; instead of using such models to identify subgroups likely to respond to specific treatments, the field relies on availability of service, trial-and-error, and clinical judgment to make treatment decisions. This is a major barrier to effective treatment of ASD. This project addresses this problem by integrating mathematical models of behavior and brain activity across adolescent development, in order to establish a neurocognitive model that can successfully predict individual adolescents' social and nonsocial learning profiles at key developmental time-points. Specifically, this work compares the suitability of various reinforcement learning models to capture selective deficits in social learning of adolescents with ASD, as well as variability in both social and nonsocial learning across typically developing (TD) adolescents and those with ASD. Identifying how these model-based predictions are implemented in brain circuits may allow for characterization of the neural architecture underlying learning in therapeutically relevant contexts. This project focuses on the understudied role of the cerebellar posterior lobe in learning processes of interest, given recent research indicating this region's involvement in updating social information. The proposed work will identify and characterize neurocognitive variability in the substrates of learning within ASD, with the long-term goal of applying these models to inform, refine, and individualize diagnosis, prognosis, education, and treatment of youth with ASD.

NIH Research Projects · FY 2025 · 2019-09

Myasthenia gravis (MG) is a chronic autoimmune disorder affecting neuromuscular transmission, with a prevalence between 77 to 167 per million. To further complicate investigations of MG is that it consists of distinct subcategories: 1) early-onset AChR antibody-positive, 2) late-onset AChR antibody-positive, 3) paraneoplastic thymoma-associated, 4) MuSK antibody-positive, and 5) AChR/MuSK antibody-negative MG. MGNet, funded in 2019 as part of the Rare Disease Clinical Research Network, aims to continue to enhance biomarker development, improve clinical trial performance, and broaden the understanding of MG pathophysiology. Despite recent FDA approvals of new therapies, treatment outcomes remain variable with significant adverse effects. Patients face challenges due to the absence of a cure, treatment complications, variable responses, high costs, and poor quality of life. Clinical outcome metrics are limited as unanimously agreed by patients, industry and investigators. No biomarkers predict or correlate with treatment response and about 10 percent of patients have no serological diagnostic marker. To address these unmet needs, we propose the following specific aims. Aim 1 will enhance clinical trial readiness through prospective patient monitoring using an electronic medical record-based application (Project 1). Aim 2 serves to identify and validate diagnostic, treatment-predictive and -responsive biomarkers to enhance early-phase clinical trials and improve disease monitoring (Projects 2 and 3). Aim 3 supports a diverse pool of investigators focusing on rare diseases and engage novel investigations into MG (Pilot Studies, Career Enhancement). Aim 4 maintains a successful Administrative Core to assure support for all investigators in their work and improve awareness among scientists, physicians, and the public regarding the unique needs of MG patients. We assure our success through our continued work with the MG Foundation of American and Conquer MG with the addition of the MG Association and the MG Foundation of Michigan.

NIH Research Projects · FY 2026 · 2019-03

Project Summary/Abstract Despite over 65 years of electroencephalography (EEG) in newborn infants, our understanding of the rhythmic cortical activity patterns associated with normal development and brain injury remains largely speculative. The long term goal of the current project is to create a developmental activity atlas in which abberant EEG patterns in the at-risk newborn are matched to specific disruptions in brain areas and neuronal types. A first step towards this goal requires an un-anesthetized preclinical model with demonstrated homology to fetal and neonatal human cortical activity in which cells can be genetically manipulated and the underlying effects on cortical circuits rigorously probed. By measuring activity in vivo through the depth of cortex and corresponding thalamic networks in developing neonatal rodents, we will provide insight into the circuit changes that may underlie human fetal thalamo-cortical development to inform future studies of subcortical injury in infants and non-human primates. By manipulating the activity of thalamic neurons we will assay the distinct contributions of the thalamic relay and inhibitory neurons to the maturation of specific features of the EEG common to developing humans and rodents. Given the challenges of imaging and diagnosis in the fragile at-risk newborn, EEG has the potential to be a valuable and inexpensive bedside diagnostic tool. An improved understanding of the control of cortical activity development will inform diagnosis after neonatal brain injury and improve targeting of treatments for the cognitive and intellectual disability that often results.

NIH Research Projects · FY 2025 · 2016-06

Obstructive sleep apnea (OSA) is recurrent closure of upper airway during sleep, which results in intermittent hypoxia (IH). OSA is the most common type of sleep disordered breathing (SDB), which is particularly prevalent in obesity affecting more than 50% individuals with BMI ≥ 30 kg/m2. OSA is a leading cause of hypertension with IH playing a major role. IH activates the sympathetic nervous system (SNS) inducing hypertension in humans and animal models. IH-induced activation of SNS originates in the carotid body (CB), a peripheral sensor of hypoxia, which provides key afferent input to medullary network that regulates SNS activity. Obesity also activates SNS and causes hypertension. CB plays a role in obesity-induced hypertension. Interactions between IH and obesity in CB, which are highly translationally relevant in SDB, have not been explored. This project will examine novel molecular targets in the CB differentially regulated by IH and obesity to optimize treatment of cardiovascular complications of OSA. During the previous funding periods of this award we identified a pathway by which obesity increases CB and carotid sinus nerve (CSN) activity leading to hypertension. Specifically, we have shown that obesity-induced hypertension is a consequence of leptin binding to the leptin receptor LEPRb on hypoxia-sensing tyrosine hydroxylase (TH) positive CB type I cells, which leads to up-regulation of the transient receptor potential melastatin-subfamily member 7 (TRPM7). Our Preliminary Data demonstrate that IH increases expression of TRPM7 in CB and CSN activity and that pharmacological and genetic blockade of CB TRPM7 abolishes IH-induced hypertension. We have shown that both leptin and IH increase CB TRPM7 gene expression via epigenetic modifications including DNA demethylation of Trpm7 gene promoter. We identified three differentially methylated regions (DMRs) in the Trpm7 promoter, which can be demethylated by 5-aza- deoxycytidine. We demonstrated that DMR region 1 (R1) is selectively demethylated by obesity via the leptin- LEPRb mechanism, while the DMR region 2 (R2) and DMR region 3 (R3) are demethylated by IH. Furthermore, we provide preliminary data that targeted methylation of these regions in vivo greatly decreases Trpm7 gene expression in CB and decreases CSN activity and blood pressure. Our overarching hypothesis is that obesity and IH interact to induce epigenetic up-regulation of Trpm7 gene expression in CB and increase CSN and SNS activity leading to hypertension. We will use our mouse models of DIO and IH, wildtype and transgenic mouse strains to address this hypothesis in four aims. Specific Aims 1 and 2 will examine how IH and obesity interact on the CB TRPM7 to induce hypertension (Aim 1) and CSN/SNS activity (Aim 2) using genetic and pharmacological blockade of LEPRb and TRPM7 in CB. Specific Aims 3 and 4 will examine how IH and obesity affect methylation of the Trpm7 promoter in CB to induce hypertension (Aim 3) and CSN/SNS activity (Aim 4) using methylated oligonucleotides complementary to the DMRs R1, R2, and R3. Our proposal will contribute to pharmacotherapy of obesity and OSA and open new horizons for translational use of epigenetics.

NIH Research Projects · FY 2025 · 2015-06

Modified Project Summary/Abstract Section The District of Columbia Center for AIDS Research (DC CFAR) was established in 2010 to galvanize HIV research in Washington, DC in response to one of the most severe urban HIV epidemics in the United States The DC CFAR has a unique citywide multi-institutional model with 260 investigator members at nine collaborating institutions: four universities - George Washington, Georgetown, Howard and American; two hospitals - Children’s National Hospital and the DC Veterans Affairs Medical Center; two community-based organizations - Whitman-Walker and Us Helping Us, People Into Living; and the DC Department of Health. The DC CFAR mission is to “Advance our multi-institutional research effort to contribute to ending the HIV epidemic and improving the health and quality of life of persons with HIV in Washington, DC and beyond in partnership with community and government.” The DC CFAR will address this mission through four Specific Aims which briefly are to: provide scientific leadership and institutional infrastructure; stimulate and support HIV research; develop the next generation of HIV investigators; and foster communication and collaboration. To address these Aims, the DC CFAR will support five Cores: the Administrative Core to provide overall leadership and management; the Developmental Core to oversee an array of pilot award and mentoring programs; and the Advanced Technology, Clinical and Population Sciences, and Social and Behavioral Sciences Cores to provide technical services, consultations and training. The DC CFAR will support two Scientific Working Groups – HIV Persistence and Cure, and Aging and Comorbidities, and one Scientific Interest Group on Women and HIV. To promote HIV research in this high priority area. The DC CFAR provides the vital infrastructure that interconnects the multisectoral HIV research response in DC, and the resources to help propel this response forward. Sustaining the commitment to the DC CFAR and its mission is essential to ending the HIV epidemic in our nation’s capital.

NIH Research Projects · FY 2025 · 2013-09

The visual pathways of the human fetal brain are highly active before birth. During this pre-visual period spontaneous activity in the retina provides the primary input to visual brain and data from neonatal rodents implicate this early activity in normal development and organization of visual pathways. While we understand much about the specialized circuits that produce activity in the developing retina, and the consequences of disruption of that activity for eye and brain outcomes, we know little of the brain activity that supports the earliest stages of sensory development. Early retinal activity is not passively transmitted in the developing brain. Rather, it is actively amplified and transformed by mechanisms unique to this developmental stage. This proposal will use a rodent model of human fetal brain development to follow the propagation and transformation of early retinal activity at each stage of the primary visual pathway in thalamus and visual cortex, to understand the mechanisms of its transformation, the role of individual thalamic regions, and the ultimate impact of this circuit on functional visual maps and responsiveness. This knowledge is important because disruption of early retinal, thalamic, or cortical activity associated with preterm birth or hypoxic birth complications can cause lasting visual impairment. Any treatment or early diagnosis (such as using EEG) requires knowledge of the normal developmental circuitry, activity and function of thalamus and cortex, which this project will provide.

NIH Research Projects · FY 2025 · 2012-09

Project Summary Liver fluke infection with Opisthorchis viverrini remains problematic in East Asia and is endemic in Thailand and Laos, where ~10 million people are infected. The public health implications of this situation are substantial since there is no stronger link between a human malignancy and a eukaryotic pathogen than that between cholangiocarcinoma (CCA, bile duct cancer) and infection with O. viverrini. Northeast Thailand reports the highest incidence of CCA worldwide, with the 2014 CCA age standardized incidence rate (ASR) of 85 per 100,000, which equates to 26,000 CCA-related deaths annually. The contrast with countries without liver flukes is stark given that incidence of CCA is less than 3 per 100,000 elsewhere (USA, 1.67 ASR, 2014). To survive in the biliary tract, the liver fluke actively releases excretory/secretory (ES) proteins and extracellular vesicles (EVs) that facilitate fluke feeding and manipulation of the host immune response, and ultimately modify cellular homeostasis that contributes to malignant transformation. Our hypothesis is that targeting proteins that are essential for several discrete facets of parasitism and/or are known carcinogens via a multi-valent vaccine will deliver a novel anti-infection/ anti-cancer therapy. We will address this hypothesis through the following aims: Aim 1. Explore the importance of key fluke ES proteins in host-parasite communication and leverage the findings to prioritize their selection as vaccine antigens. Aim 2. Determine whether mRNA and/or protein subunit vaccines induce antibodies that reduce both fluke burdens and CCA incidence in the hamster infection-cancer model. This proposal targets liver fluke ES proteins which drive the phenotypic hallmarks of cancer in the biliary epithelium: 1) the granulin-like growth factor, Ov-GRN-1, known to induce rampant cholangiocyte proliferation; 2) the tetraspanin, Ov-TSP-2, a key element of EVs and host-cell communication; 3) the Ov-catF cysteine protease, a key component of the protein digestion cascade; and 4) the Ov-M60mucinase which degrades host defensive mucus in the bile ducts. The conceptual innovation we utilize is a rodent model of human carcinogenesis where liver fluke infection is a known risk factor. Technical innovations include gene knockout in the liver fluke, targeting key pathogenic and nutritional processes, and combining the findings to develop vaccination against fluke infection and the infection-induced cancer. These studies will determine whether fluke proteins that communicate at the host-parasite interface represent the Achilles’ heel of this carcinogenic parasite. Targeting fluke-host communication in combination with nutrient acquisition pathways may ultimately combat liver fluke-induced bile duct cancer in the form of an anti-fluke, and indeed anti-cancer vaccine, a public health development with the potential to benefit millions of (often impoverished) residents of endemic regions in East Asia.