Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY / ABSTRACT Coronavirus disease 2019 (COVID-19) is an ongoing global pandemic. Despite substantial short term mortality risk, the overwhelming majority of infected patients survive acute illness, resulting in a growing population at risk for long term events. Cardiopulmonary symptoms are common after COVID-19, as shown by survey data reporting fatigue (53%), dyspnea (43%), and worsened quality of life (44%) 60 days after acute infection, but mechanism and time course of symptoms are unknown. Recent studies and our own preliminary data have shown myocardial tissue abnormalities on cardiac magnetic resonance (CMR) to be common in COVID-19 survivors – raising the possibility that symptoms stem from viral effects on the heart. However, CMR findings to date are limited by small size and clinical data susceptible to referral bias, raising uncertainty as to generalizability. It is also unknown whether altered myocardial tissue properties (fibrosis, edema) impact clinical outcomes. The central hypothesis of our research is that CMR tissue characterization will be incremental to clinical assessment and cardiac contractile function for prediction of long-term cardiopulmonary symptoms, effort tolerance, and prognosis among COVID-19 survivors. To test this, we will study patients from an active multi- ethnic NYC registry of COVID-19 survivors: We have already leveraged echocardiographic imaging data from this registry to show that (1) adverse cardiac remodeling (dilation, dysfunction) markedly augments short term mortality, (2) COVID-19 acutely alters left and right ventricular remodeling, and (3) many patients who survive initial hospitalization for COVID-19 have adverse cardiac remodeling – including 40% with left ventricular (LV) dysfunction and 32% with adverse RV remodeling (dilation, dysfunction): Our current proposal will extend logically on our preliminary data to test whether CMR tissue characterization provides incremental predictive utility with respect to reverse remodeling and prognosis. At least 510 COVID-19 survivors will be studied. Echo will be analyzed at time of and following COVID-19 for longitudinal remodeling, as will CMR at pre-specified (6- 12, 36 month) follow-up timepoints. Established and novel CMR technologies will be employed, including assessment of cardiac and lung injury, high resolution (3D) myocardial tissue characterization, and cardiopulmonary blood oxygenation. In parallel, QOL, effort tolerance (6-minute walk test), biomarkers, and rigorous follow-up will be obtained to discern clinical implications and relative utility of imaging findings. Aim 1 will identify determinants of impaired quality of life and effort intolerance among COVID-19 survivors. Aim 2 will test whether myocardial tissue injury on CMR is associated with impaired contractility, and whether fibrosis predicts contractile recovery. Aim 3 will determine whether myocardial tissue injury is independently associated with adverse prognosis (new onset clinical heart failure, hospitalization, mortality). Results will address key knowledge gaps regarding COVID-19 effects on the heart necessary to guide surveillance, risk stratification, and targeted therapies for millions of COVID-19 survivors at risk for myocardial injury, cardiopulmonary symptoms, and adverse prognosis.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Cardiovascular disease (CVD) is the most common non-communicable disease and the leading cause of morbidity and mortality in Haiti. It has surpassed infectious diseases over the past decade. Hypertension (HTN) is the single most common CVD risk factor in Haitian adults and its onset is 10-20 years earlier than among Black Americans in the US. Potential drivers of HTN in Haiti include social determinants such as extreme poverty, food insecurity, a high salt diet, psychological stress, and also environmental pollutants such as high lead exposure. Concurrently, Haiti continues to have the highest HIV prevalence in the region with over 35% of persons living with HIV having HTN. To curb this alarming CVD epidemic, Haiti urgently needs CVD research training for both young investigators and research staff in order to increase its capacity to conduct interdisciplinary, impactful CVD research. This proposal builds on a ~40-year history in Haiti of NIH research and Fogarty training by the Groupe d’Etude du Sarcome et des Infections Opportunistes (GHESKIO) and its US collaborating partner, Weill Cornell Medicine. The Cornell-GHESKIO partnership began with a focus on HIV and since 2015 has expanded to address the CVD epidemic in Haiti. We have built a strong foundation with 8 active NIH and foundation grants in CVD research and the creation of the GHESKIO NCD Clinical and Research Unit. We now need to train a skilled workforce of specialized CVD researchers who can expand Haiti’s research portfolio and transform research findings into practice. The goal of this training program is to strengthen research that will ultimately improve CVD-related health outcomes in Haiti and the world. Our three training objectives include: 1) to provide long-term mentored CVD research training to 8 Haitian clinician scientists through MPH coursework at Quisqueya University with mentored CVD research in Haiti; 2) to train 24 medium-term research staff in CVD knowledge and technical research skills through a new 1-year CVD Core Research Course taught by Haitian and international faculty; and 3) to establish a CVD Implementation Network of 6 CVD clinics to increase the uptake, optimization, and evaluation of high-impact CVD interventions in Haiti. Staff at the Network clinics will have opportunities for short-term training including participation in monthly CVD Grand Rounds and participation in 1-week workshops within the CVD Core Research Course. In addition, we will sponsor an annual national CVD Research Conference with a focus on trainee leadership and career development. The proposed program is the first CVD research training program in Haiti and brings innovation in CVD coursework, new research mentors and faculty, and a platform for building implementation capacity for CVD interventions across the country. We are committed to training the next generation of CVD researchers in Haiti.

NIH Research Projects · FY 2025 · 2021-09

Accumulating data from human and mouse support the hypothesis that system level lipid disregulation is an early and critical factor in etiology and progression of Alzheimer's disease (AD). The explosion of 'omics methods in the past decade has resulted in a proliferation of various studies and data sets that interrogate specific regions of the brain. Using Imaging Mass Spectrometry (IMS), our preliminary studies have found regionally differential lipid composition in coronal sections from wild type mouse brain and a mouse model of Alzheimer's disease over expressing the amyloid precursor protein (APP) transgene. This regional lipid disregulation requires system-wide interrogation of lipid homeostasis which can singularly be accomplished with lididomics. A candidate based screen of lipid modifying enzymes in mouse embryonic stem cells for resistance to Aβ-triggered synapse loss, identified multiple metabolic enzymes which may be responsible for exit of polyunsaturated fatty acids (PUFA), specifically docosahexaenoic acid (DHA) from an acyl chain remodeling pathway, the Land's cycle. The Land's cycle has recently been identified to be dysfunctional in two animal models of AD. In the human context, DHA transport into the brain is aberrant by age 30, in carriers of a variant of apolipoprotein E (ApoE4) strongly associated with AD risk. Synthesis of these results with multiple hits from GWAS implicating lipid metabolism and transport, strongly support system-wide dyshomeostatis of acyl chain composition in the brain. However, the reports of regionally defined lipid composition are currently limited. We propose to test the hypothesis that acyl chain composition among multiple lipid classes is severely disregulated in brain regions known to be susceptible in AD including hippocampus and entorhynal cortex. Using IMS we will interrogate the lipid composition of mouse models of AD, Tg2576 and targeted replacement APOE mice as well as human brain tissue. We will then test the hypothesis that DHA accretion is a critical modifier of AD associated behavioral deficits and pathology in mouse models using knock-out lines of acyl- CoA synthetase 6 (Acsl6), a key mediator of DHA enrichment in the brain. We will generate new strains of Tg2576 and TRAPOE4 lacking Acsl6 and overexpressing Acsl6 to determine both necessity and sufficiency of DHA brain accretion for AD associated deficits. Finally, we will integrate and assemble our data into a publically available lipid brain atlas. These studies have potential to synthesize accumulating lipidomics data in aging and neurodegenerative disease. The use of spatial lipidomics at scale in the brain has not been yet be comprehensively accomplished, and is required for clear understanding of the basic metabolic pathways thus uncovering connectivity and functionality in the brain. Completion of these studies will represent compelling evidence for the critical nature of lipid composition in basic biology of AD and lead to new strategies for biomarker discovery as well as therapeutic targets.

NIH Research Projects · FY 2025 · 2021-09

Leveraging linked registry and electronic health records to examine long-term patient outcomes after peripheral vascular intervention Project Summary/Abstract Peripheral arterial disease (PAD) affects over 200 million people worldwide. Peripheral vascular interventions (PVI) are the most common procedures that are performed to manage PAD. Existing randomized controlled trials (RCTs) and observational studies of patient outcomes after PVIs all had limited follow-up lengths due to difficulties in long-term data collections. In addition, heterogeneity of treatment effect (HTE) for stent placement vs. percutaneous transluminal angioplasty (PTA) alone has not been well understood with the current approach of effect modifier assessment. Real-world data (RWD), particularly registries linked with electronic health data (EHR), are useful for studying long-term outcomes after vascular procedures. However, methods for working with multiple data sources and analyzing unstructured text data are still evolving. The proposed research aims to address current evidence gaps in long-term patient outcomes after PVI procedures. This will be facilitated by innovatively apply and refine data linkage, natural language processing (NLP), and effect modifier assessment methods. Specifically, this project will link registry and EHR data to 1) examine long-term major adverse limb events after stent placement vs. PTA alone as well as assess heterogeneity of treatment effect by patient characteristics; 2) develop an NLP pipeline with machine learning methods to analyze unstructured text data and examine long-term efficacy endpoints after stent placement vs. PTA alone, and; 3) establish feasibility and updating requirements for the deployment of the NLP tool for long-term PVI outcome assessment to other institutions. To support the research activities and the transition toward independence, the candidate will undertake the following career development activities during the award period: 1) gaining an in- depth understanding of NLP and machine learning methods; 2) refining data science expertise to integrate EHR into medical device epidemiologic research; 3) strengthening knowledge in current and novel vascular disease treatment; 4) developing and improving skills in grant writing and academic leadership; 5) training in responsible conduct of research. The candidate will be mentored by a team of experts with complementary strengths in surgical and device outcomes research, natural language processing and machine learning, and vascular disease and surgery. The proposed career development and research activities will develop the candidate's skillset and expertise and lead to an R01 level application. The candidate's long-term goal is to become an independent researcher focusing on the development and application of advanced multidisciplinary methods in the evaluation of surgical and device outcomes in the vascular disease area, supporting clinical, patient, and regulatory decision-making.

NIH Research Projects · FY 2025 · 2021-08

Modified Project Summary/Abstract Section Despite the success of standard antiretroviral therapy (ART), the need for an HIV cure remains compelling, both to improve the lives of PWH and to bring about the end of the pandemic. Strategies for an HIV cure fall under two categories: those that seek ART-free ‘remission’, and those targeting a classical cure or ‘eradication’. While precedents exist for both scenarios, the latter have only been achieved with bone marrow transplantation. In contrast, although naturally occurring immune-mediated control of HIV (remission) is relatively rare, many such cases have been described. Our proposed “Martin Delaney Collaboratory for HIV Cure Research” program is entitled “REACH” - Research Enterprise to Advance a Cure for HIV. The central theme of REACH is that cellular immune responses (NK and T-cells), combined with next generation virus-neutralizing biologics, can be harnessed to achieve durable remission and eradication of HIV reservoirs. The proposed research focuses on closing gaps in our understanding of the fundamentals of the system that we are trying to perturb, i.e.: the HIV reservoir in relation to cellular immunity, as the means to achieve real progress towards effective and viable HIV cure strategies. Our approach centers around three research foci, which emphasize back to basics science, but connect this with discovery to translational pipelines directed towards both remission and eradication. The proposed objectives, broadly defined, aim to: (1) redefine the three-way relationship between the persistent HIV reservoir, CD8+ T-cells, and rebound virus at the levels of: single cells, individuals, and populations, (2) harness conventional and unconventional (bNAb-induced) CD8+ T-cell responses, in combination with bNAbs and ‘next generation’ biologics, to achieve durable control of HIV replication, and (3) develop a discovery-to-translation pipeline to overcome multiple barriers to the eradication of HIV reservoirs by CTL/NK cells. These studies will be rooted in a strong basic science program that will contextualize results with novel insights into barriers to immune-mediated reservoir elimination, including the role of the proviral integration site and of viral and host factors influencing immune susceptibility. These objectives will be realized by a group of accomplished investigators with strong collaborative histories, along with community and industry partners.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY/ABSTRACT The candidate’s career goal is to become an independent physician-scientist studying HIV and aging, and the increased burden of medical co-morbidities and geriatric syndromes in older adults with HIV (OAH). The candidate has laid the foundation for achieving this goal by gaining clinical expertise in caring for OAH, conducting research, and obtaining a Master’s Degree in Clinical and Translational Investigation. To achieve her career goal, the candidate will need to expand upon her geroscience and translational research skills, including additional biostatistics and research methods training which are described in this resubmission. A key component of the candidate’s training will be conducting the proposed research project. Despite effective antiretroviral medications, OAH bear a greater burden of medical co-morbidities and geriatric syndromes than their HIV-negative peers. Translational research investigating biomarkers inflammation offers as opportunity for insight to the process of accelerated/accentuated aging that is observed in OAH. Cell-free mitochondrial DNA (cfmtDNA) is released from cells undergoing stress and necroptosis-mediated cell death and has the potential to serve as a mediator and marker of chronic immune activation and dysregulation. We hypothesize that cfmtDNA will be associated with lower cognitive performance and greater frailty in a longitudinal study of OAH. Previously, we have studied a cohort of OAH (age 55 and over) at our institution, and those with cognitive impairment had higher average levels of cfmtDNA in plasma than participants without cognitive impairment. We propose to leverage this existing study to investigate the following specific aims: 1) Determine the association between cfmtDNA and cognition in OAH; 2) Determine the association between cfmtDNA and longitudinal physical function in OAH; and 3) Evaluate the immunostimulatory potential of cfmtDNA from OAH with and without cognitive decline. Participants from our existing cohort will be invited back for two study visits separated by 18-24 months, each visit will include detailed neurocognitive assessment, physical function measures, falls and instrumental and activities of daily living, and blood and urine specimen collection for analysis and creation of a biorepository. Together, these investigations will shed light on the relationship between cfmtDNA, immune activation and geriatric-related syndromes in OAH. This project proposes a five-year, multifaceted training program under the mentorship of Dr. Marshall Glesby as the primary mentor, as well as Drs. Mary Choi, Lishomwa Ndhlovu, and Eugenia Siegler as co-mentors. Together with a Scientific Advisory Committee, they will provide the expertise in research design, biomarkers, immunology and geroscience that will allow support the success of this project. The completion of the proposed project will lead to an enhanced understanding of cfmtDNA as a biomarker of geriatric syndromes in OAH, and a translational research tool to identify OAH at the highest risk of morbidity and mortality. After completion of this project, the candidate will be poised to submit a competitive R03 and R01 proposals.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY/ABSTRACT: Sepsis is a deadly infection characterized by a dysregulated host immune response. Outcomes have failed to improve despite decades of research. The immune response in sepsis is varied. Immunologic therapy has failed in part due to the heterogeneity of the syndrome. Beyond the immunologically silent apoptosis, necroinflammatory cell death, commonly necroptosis, is immunologically stimulating and can perpetuate inflammation in sepsis. The initiation and coordination of necroinflammatory cell death is complex. TNF related apoptosis inducing ligand (TRAIL) coordinates cellular processes associated with increased apoptosis and necroinflammatory cell death. Receptor interacting serine/threonine kinase 3 (RIPK3) is essential to necroptotic cell death. Our work has shown that RIPK3 is increased in septic patients in the intensive care unit in parallel with increased organ dysfunction and is associated with poor outcomes. In the ICU, we have demonstrated that lower TRAIL is associated with higher RIPK3 and increased organ dysfunction. In this project, we will examine TRAIL and RIPK3 at three time points, in the emergency department and ICU. We hypothesize that necroinflammatory cell death, characterized by high RIPK3 and low TRAIL will identify those who progress to sepsis and septic shock and that there will be novel patterns of necroinflammatory cell death in patients at increased risk of death with sepsis. AIM 1 will create a human cohort of patients at three critical time points during an acute admission to the hospital. The first is soon after admission to the emergency department prior to resuscitation and the administration of antimicrobial therapy. The follow up blood draws are obtained following admission to the ward or ICU when organ dysfunction is established and therapy has been initiated. AIM 2 will examine the relationship between TRAIL and RIPK3 and sepsis, septic shock and mortality through two methodologies. The first will examine whether TRAIL and RIPK3 will increase our ability to diagnose sepsis when combined with physiologic sepsis prediction tools in the emergency department. The second will evaluate the effect of the follow up TRAIL and RIPK3 on outcomes, after modeling the effect of time dependent patient, pathogen and treatment factors. For AIM 3, we will measure levels of a targeted mechanistic cell death panel including, RIPK1, RIPK3, MLKL, along with key damage associated molecular patterns, mtDNA and HMGB1. We will also evaluate a broader necroinflammatory biomarker panel in a proteomics platform. We will then evaluate whether there are clusters of patients defined by relative biomarker levels together with physiologic variables. We will examine if these patient clusters have differential outcomes. If the aims of this proposal are achieved, we will have useful information concerning the role of necroinflammatory cell death in human sepsis from multiple time points. Results from this study may offer insight into the development of biomarkers for predictive enrichment of clinical trials targeting necroinflammatory cell death.

NIH Research Projects · FY 2025 · 2021-08

This MPI R01 combines the expertise of the Departments of Psychiatry, Neurology, and Internal Medicine at Weill Cornell Medicine to address MCI (mild cognitive impairment) and early stage AD/ADRD in patients with comorbid chronic pain and depression. We propose to test whether Problem Adaptation Therapy for Pain (PATH-Pain), a novel primary-care based psychosocial intervention designed to reduce stress in MCI and early stage AD/ADRD patients with comorbid depression and pain, has better cognitive, affective, and functional outcomes than Attention Control Usual Care. PATH-Pain is an easy-to-administer psychosocial intervention designed to improve emotion regulation and reduce stress in older adults with MCI or early stage AD/ADRD, chronic pain, and depression. To reduce stress, PATH-Pain aims to: a) reduce negative emotions associated with pain and pain-related disability; b) reduce negative emotions that interfere with pain treatment (e.g., hopelessness, helplessness); c) increase positive emotions and increase engagement in pleasurable activities; d) help patients identify addressable problems in their lives and try to find the best possible solution to these problems; e) reduce interpersonal tension between patients and family members, caregivers, and friends; and f) shift attention during experiences of pain to reduce pain intensity. To achieve these aims, PATH-Pain employs emotion regulation, problem solving, and behavioral activation techniques. In our preliminary study, PATH-Pain participants showed high acceptability and treatment satisfaction with the intervention. Based on our power analysis, we will randomize 100 older adults (60 years and older) with MCI or early stage dementia (probable or possible Alzheimer’s Disease), comorbid depression, and chronic pain to 8 weekly in-office sessions and 6 monthly phone (3 individual and 3 group) sessions of PATH-Pain vs. Attention Control Usual Care in 4 primary care sites. The Attention Control Usual Care arm consists of Usual Care, a pamphlet on pain and depression, and a structured interview of the same duration as each PATH-Pain session. The structured interview aims to control for attention and time, and will consist of general questions regarding health habits and other non-medical topics unrelated to cognitive impairment, pain, and depression. The investigators have shown evidence of feasibility of recruitment, retention, and assessment procedures for the proposed study. Assessments will be performed at baseline, and weeks 5 (no cognitive outcomes), 9, 24 (no cognitive outcomes), 36, and 52 by research assistants unaware of the study hypotheses and the participants’ randomization status. PATH-Pain will be administered by certified mental health workers and the Attention-Control sessions will be administered by a non-clinician team member experienced in structured interviews.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY/ABSTRACT Obesity-induced lipotoxicity is the primary pathophysiological defect that predisposes to non-alcoholic fatty liver disease (NAFLD). Because current management options remain limited, identification of new regulatory mechanisms that govern the maladaptive response to overnutrition will serve to identify novel opportunities for pharmacologic intervention. Acyl-CoA thioesterases (Acots) control the cellular utilization of fatty acids by hydrolyzing acyl-CoA into non-esterified fatty acids. Our long-term goal is to define how aberrant Acot activity can be leveraged for therapeutic purposes. The objective of this research is to determine the mechanisms by which Acot9-mediated hydrolysis of acetyl-CoA culminates in metabolic disease. Our preliminary data indicates that Acot9 locates to the inner membrane (IM) of the mitochondria in the liver where it traffics acetyl-CoA towards the citric acid (TCA) cycle. This results in increased hepatic glucose production (HGP) and de novo lipogenesis (DNL) as well as lipotoxicity as evidenced by reactive oxygen species (ROS) and hepatic insulin resistance. In addition to TCA cycle, Acot9 increased acetyl-CoA supply for lysine acetylation of proteins (AcK) by controlling acetyl-CoA bioavailability and by inhibiting the deacetylase sirtuin 3 (Sirt3), which reduces ROS by inhibiting AcK. In contrast to liver, in thermogenic adipose tissue (BAT), cold-induced translocation of Acot9 from IM into mitochondrial matrix suppressed thermogenesis by trafficking acetyl-CoA away from TCA cycle. Our central hypothesis is that obesity-induced activation of Acot9 impairs nutrient homeostasis by promoting lipotoxicity in the liver and by limiting thermogenesis in BAT. The rationale is that the mechanisms of acetyl- CoA trafficking by Acot9 will reveal novel targets for the prevention of lipotoxicity and its pathophysiological consequences. The central hypothesis will be tested in three specific aims: 1) To identify the molecular mechanisms by which hepatic Acot9 promotes hepatic lipotoxicity; 2) To elucidate the mechanisms by which Acot9 in BAT limits thermogenesis; and 3) To determine the mechanisms by which Acot9 controls AcK and ROS in the liver. In Aim 1, the mechanisms of Acot9-induced lipotoxicity, HGP and DNL will be elucidated in mice with liver-specific ablation of Acot9 (Acot9LKO) using lipidomics and metabolomics. Impact on insulin signaling and lipotoxic pathways will be determined in mice and primary hepatocytes. Aim 2 will use mice with BAT-specific ablation of Acot9 for the indirect calorimetry measurements in climate-controlled cages. Clamp, tissue histology and metabolomics will assess the metabolic function of Acot9 in BAT. Cultured brown adipocytes and recombinant Acot9 will be used to determine the mechanism of Acot9 translocation into mitochondrial matrix. Aim 3 will use Acot9LKO/Sirt3–/– double knockout mice to determine the role of Sirt3 in Acot9-mediated regulation of AcK and ROS in the liver. Overall, this proposal will elucidate new mechanisms of thioesterase-mediated control of acetyl-CoA utilization. This is significant because acetyl-CoA is the common breakdown end-product of nearly all dietary lipids and sugars. These studies are expected to establish Acot9 as a tractable target for the management of NAFLD.

NIH Research Projects · FY 2026 · 2021-08

Longitudinal Analysis of Diffusion Tensor Imaging to Discover Adolescent Alcohol Use Effect PROJECT ABSTRACT Alcohol abuse is the third leading preventable cause of death in the United States. A signature injury of Alcohol Use Disorder (AUD) is in the white-matter (WM) microstructure and its constituents, which enable connectivity of proximal and distal brain structures and functional integration. Despite the progress in understanding alcohol’s effects in adults, still unclear is the causal direction between abnormal WM maturation and initiation of heavy yet non-dependent drinking during adolescence. To gain insight into regional development of connectivity, the National Consortium on Alcohol and NeuroDevelopment in Adolescence (NCANDA), a prospective longitudinal study of 831 youth to track normal and alcohol-related deviant neurodevelopmental trajectories, acquired diffusion tensor imaging (DTI) data annually. Unraveling alcohol effects from the complex dynamic course of adolescent neurodevelopment requires highly sensitive analysis approaches. In this project, I propose a new type of longitudinal DTI analysis, in which a unified trajectory model is used both for explicitly capturing biologically-plausible variation in DTI measurements across visits within each subject and for estimating a group-level developmental trajectory. Such an approach will result in longitudinally consistent DTI measurements enabling accurate characterization of microstructural development in normal adolescents. Based on this approach, I will test the hypothesis that precursors of drinking onset and the disruption effects induced by initiation of alcohol use are related to different brain regions. The analysis will identify the precursors by comparing developmental trajectories prior to drinking onset between the no-to-low and heavy drinking cohorts and will identify the disruption by comparing trajectories after drinking onset. The alcohol- induced disruption to WM maturation will be further stratified with respect to age, highlighting the heightened vulnerability in younger adolescents. Lastly, the proposed longitudinal analysis also facilitates further tracking of microstructural remodeling following abstinence to identify reversible or persistent alcohol-related injury. Revealed imaging phenotypes linked to adolescent heavy drinking could point to potential causes and precursors of AUD, which could in turn improve diagnosis and prevention in clinical settings. Moreover, researchers will be able to use the proposed longitudinal approach as a general tool to answer their neuroscientific questions in the context of seeking developmental change.

NIH Research Projects · FY 2024 · 2021-08

PROJECT SUMMARY Nearly 2 million people are diagnosed with lung cancer each year and it is the leading cause in cancer related death worldwide. Standard of care for non-small cell lung cancer (NSCLC) patients has changed little over the past several years however targeted therapies and immune checkpoint inhibitors have recently become a promising option. The most common oncogenic drivers in NSCLC are mutations in EGFR and KRAS which upregulate a myriad of downstream signaling pathways that promote tumor cell growth. Until recently, efforts to develop therapeutics that directly target EGFR or KRAS have largely been met by failure. Specifically, development of farnesyl transferase inhibitors and downstream pathway inhibitors like MEK and RAF, have not shown improvement in survival in the clinic and resistance mechanisms are described. The discovery of KRASG12C mutant inhibitors like AMG 510 and MRTX849 is encouraging as they have both shown promising signs of clinical activity and promise to transform treatment of KRAS mutant cancer. These inhibitors work by covalently binding to the reactive Cys12 locking KRAS in its inactive GDP-bound state. However, as with previously targeted therapies, mechanisms of resistance are beginning to be described. Utilizing precision modeling in mice, I will test the hypothesis that KRAS allelic imbalance and genetic determinants in NSCLC drive tumor progression and confer unique responses to targeted therapies. Since our lab has developed LSL-Kras allelic series that allows for selective targeting of the WT Kras allele, in Aim 1, I will use CRISPR-based genome editing technology to knockout WT Kras in vivo and measure effects in tumor burden and G12C inhibitor response. Further, I aim to understand how WT KRAS signaling contributes to the tumor immune microenvironment and how it affects targeted treatment response. In Aim 2, I will use a patient data guided approach to elucidate how cooperative mutations in tumor suppressors effect tumor progression and G12C inhibitor response. This powerful genetic approach will allow me to directly interrogate ways in which KRASG12C targeted therapy can be affected. Identifying an effective approach to disrupt KRASG12C mutant NSCLC will have a profound impact on the clinical management of these patients. Thus, we believe our work will contribute significant pre-clinical data to developing safe and effective targeted therapies for NSCLC and other cancer types.

NIH Research Projects · FY 2024 · 2021-08

Abstract Since 2010, clinical medicine has benefited from a rapid surge of clinical research on chronic diseases using data from electronic health records (EHRs). EHRs are appealing because they can offer large sample sizes, timely information, and a wealth of clinical information beyond that obtained from either health surveys or administrative data. However, while millions of patient records are included in large EHR records, they are not population-representative random samples, a constraint that potentially biases inferences based on such data and, therefore, has limited their utility for population health research. EHR data typically contain multiple types of biases, particularly: 1) sampling inclusion bias: EHR data only include information on patients visiting participating medical systems, and they primarily capture data when patients are ill. Even among populations with a particular disease, patients represented in EHRs tend to over-represent individuals who are sicker and have higher health care utilization; 2) sampling frequency bias: the numbers of patients’ encounters and features in EHRs are at various frequencies and these frequencies correlate with both patients’ characteristics and outcomes; and 3) institution bias: EHR samples of any hospital reflect the characteristics of patients population served by that specific hospital. Consequently, EHR-based risk prediction models will have 1) biases in risk factor selection and estimation for population inferences; 2) disparate mistreatment (unfairness) in terms of variation in a model’s prediction accuracy across patient subgroups (such as gender, race, and age) with various sampling inclusion probabilities or frequencies; 3) biased prediction model to reflect characteristics of patients served by the local hospitals. We propose to develop: 1) effective sample-weighting method to correct biases in risk factor selection and estimation for population inferences (Aim 1), 2) flexible deep learning method for EHR personalized risk prediction with fairness criteria (Aim 2); and 3) innovative calibration method to improve reproducibility of EHR-based risk models between institutions (Aim 3). We will predict risk of subsequent incident cardiovascular disease (CVD) in patients with type 2 diabetes (T2DM) as a demonstration of methodology development. Broader use of these methods will be generally applicable to other diseases outcomes and population of interest. To develop and validate these methods, we propose to analyze three unique datasets: 1) the New York University Langone Health EHR data (NYU-CDRN, 2009 to now) including demographics, vitals, diagnoses, lab results, prescriptions, and procedures; 2) the New York City Clinical Data Research Network (NYC-CDRN)—an EHR network comprising 20 NYC healthcare institutions, including the NYU-CDRN, with longitudinally linked data on >12 million patient encounters under a Common Data Model, and 3) the Health and Retirement Survey (HRS, begun in 1992 and ongoing), as a benchmark population- based cohort, that has nationally representative health interview data for over 20 years, as well as biomarkers, physical assessment information, prescription drug data, and claims linkages.

NIH Research Projects · FY 2024 · 2021-08

PROJECT SUMMARY/ABSTRACT Staphylococcus aureus is a major cause of both community-acquired and nosocomial infections, which have become increasingly challenging to treat due to the widespread evolution of antimicrobial resistance. There is a critical need for the development of alternative therapeutic approaches against multidrug-resistant bacteria that also spare the protective commensal microbiota, which often provide colonization resistance against pathogens. Bacterial disease is driven by S. aureus toxins and other virulence factors, which are mainly encoded by mobile genetic elements (MGEs). In particular, numerous enterotoxins and the superantigen toxin causing Toxic Shock Syndrome are all carried by a class of MGEs called the S. aureus Pathogenicity Islands (SaPIs), which spread between bacteria by hijacking the reproductive machinery of bacteriophages. Staphylococci also possess CRISPR-Cas systems, which provide adaptive immunity by blocking invading MGEs like phages and plasmids. In this proposal, building upon preliminary data, I will test the central hypothesis that CRISPR-Cas systems also prevent the transmission of SaPI elements and their associated virulence genes. In Aim 1, I will define the complex tripartite interplay between staphylococcal CRISPR systems, SaPIs, and their helper phages using various molecular and genetic approaches. In Aim 2, I will investigate the mechanisms by which SaPIs manage to overcome CRISPR-mediated restriction and disseminate throughout bacterial populations. I anticipate that these studies will elucidate both the molecular basis and biological consequences for CRISPR-SaPI interactions. In Aim 3, I will evaluate whether CRISPR can be used to selectively kill SaPI-harboring S. aureus and establish a proof-of-concept for CRISPR-based antimicrobials directed against virulence-encoding MGEs. The proposed experiments will contribute to the long-term goal of designing alternative therapeutic approaches in an effort to overcome the shortcomings of antibiotics in treating multidrug-resistant infections. This fellowship will support my training in the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, including my doctoral work in the laboratory of Dr. Luciano Marraffini at Rockefeller and the remainder of my medical training at Weill Cornell. The training plan outlined in this fellowship project is designed to optimally prepare me for a research career as an independent principal investigator and physician-scientist.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Alzheimer's disease (AD) is the most common form of dementia in elderly. Amyloid-β (Aβ) and tau pathologies and neuroinflammation are three major hallmarks of Alzheimer's disease. Vast majority of the drug discovery efforts in the past decades have focused on targeting the Aβ pathology, but none are successful in Clinical Trials. While tau pathology, not the Aβ pathology, has emerged to play critical role in memory decline in AD, drugs targeting the direct effects of tau on neurons also have not met success either. Compelling human genetic studies link the innate immune responses to elevated risk of developing late-onset AD, supporting targeting microglia, resident immune cells in the brain, as the next-generation treatment for AD. We showed that a critical role of cyclic GMP-AMP synthase (cGAS)-Stimulator of interferon genes (STING) signaling in microglial toxicity and tau-mediated cognitive decline. Activation of cGAS, a major cytosolic dsDNA sensor, catalyzes production of cGAMP, an extremely potent STING agonist as the second messenger that activates cGAS-STING pathway, leading to a production of the antiviral responses through activation of interferon regulatory factors (IRFs) and expression of cytokine and type I interferon genes. We found that a partial or complete genetic cGAS ablation protected against the tau-mediated spatial learning and memory deficits in PS19 Tau mice. Moreover, treatment with a small molecule inhibitor of cGAS reduces interferon responses, diminished microgliosis, and protected against cognitive deficits in an AD mouse model with tauopathy. We hypothesize that inhibitors the cGAS activity will dampen neuroinflammation and maladaptive immune responses, protect against AD-related deficit. We propose to develop small molecule human cGAS (h-cGAS) inhibitors as novel microglial modulators to treat AD. In Aim 1, we will develop lead h-cGAS inhibitors starting with two known hits and determine whether these inhibitors effectively modulate the cGAS-STING pathway in cell-free and cell-based assays. We expect to identify new hits via hits expansion and synthesize >200 analogs. Aim 2 focuses on optimization of analogs a potent cGAS inhibitor, TDI-6570, which is a lead low nanomolar potent mouse cGAS (m-cGAS) inhibitor and possesses 10x less h-cGAS activity. We will design up to 50 new analogs. Results of SAR, docking experiments, and in- silico calculation will be used to maximize the lead quality. Completion of Aim 1 and Aim 2 will lead to 5 lead compounds for in vivo and efficacy studies. In Aim 3, we will establish PK and efficacy of h-cGAS inhibitors in mouse model of tauopathy, and efficacy in human stem cell-derived microglia and cerebral organoids with tauopathy. At the end of the proposed 5 years study, we anticipate identifying 1-2 lead h-cGAS inhibitors as tool compounds and a proof of principle to further advance as drug candidates to treat Alzheimer's disease and related neurological disorders.

NIH Research Projects · FY 2024 · 2021-08

PROJECT SUMMARY The colon is a major segment of the intestine and differs significantly from the small intestine in morphology, cell types, physiological function and disease susceptibility. Devastating and prevalent diseases, including colorectal cancers and ulcerative colitis, arise from colon but not small intestine. Colon absorbs water but cannot uptake most nutrients like the small intestine and consequently a significant loss of the small intestine will lead to digestive failure that the colon cannot compensate. Despite significant progress, aspects of the colon biology remain poorly understood. Molecular determinants that distinguish the colon from the small intestine and govern colon-specific cell lineage differentiation and homeostasis remain largely uncharacterized, hindering a deeper understanding of regionalized intestinal diseases. In preliminary studies, we identified SATB2, a chromatin factor with restricted expression in the colonic epithelium, as a crucial molecular regulator of colon identity and differentiation. SATB2 deletion from adult intestine led to a homeotic-like transformation of colonic epithelium into one that resembles small intestine ileum in cellular composition and gene expression, and the mutant colon can absorb nutrients, a function unique to the small intestine. These data suggest that SATB2 is a potential “master regulator” of colonic epithelium. The identification of SATB2 offers a unique opportunity to study colonic ontogeny and fate determination, and assess its therapeutic implications. In this project, Aim 1 will evaluate the hypothesis that colonic stem cells harbor primed ileal enhancers and thus harbor a chromatin-level permissiveness for ileal transcriptional activation and cell fate plasticity. Aim 2 studies will evaluate the hypothesis that SATB2 recruits two chromatin remodeling factors, MTA2 and SMARCD2, to separate pools of colonic and ileal enhancers to modify local chromatin, allowing differential access of intestinal transcription factors and effecting transcriptional regulation. In Aim 3, using mouse models of Short bowel syndrome (SBS), we will evaluate whether promoting colonic nutrient absorption can combat digestive failure and the associated pathophysiology in SBS. These studies together will elucidate the cellular and molecular mechanisms by which SATB2 preserves colonic identity and effects a colonic to ileal conversion, which may be exploited as a novel therapeutic approach to treat SBS.

NIH Research Projects · FY 2025 · 2021-07

Abstract The human heart shows little regenerative capacity following an injury such as myocardial infarction (MI). Instead, the heart scars, decreasing cardiac function, and leading to heart failure. There is no clinically meaningful regenerative therapy available for MI patients. By contrast, adult zebrafish regenerate heart muscle after severe cardiac damage without significant scarring. This is achieved through proliferation of existing cardiomyocytes (CMs), aided by the environment provided by non-muscle cells, such as the epicardium, a mesothelial cell sheet covering the surface of the heart. An analogous regenerative machinery of CM proliferation and epicardium contributions also exists in the adult mammalian heart; however, it is not sufficiently activated for significant regeneration. Recent studies demonstrated that restoring epicardial factors through the application of epicardial patches or co-transplantation of human stem cell-derived epicardial cells together with stem cell- derived CMs after an MI benefit heart regeneration. Thus, enhancing the pro-regenerative activation of the epicardium may benefit mammalian heart regeneration after MI. We and others previously found that the zebrafish epicardium is activated by injury and aids muscle regeneration through paracrine effects and as a source of multipotent cells. However, little is known about the cellular and molecular mechanisms controlling epicardial activation that lead to successful heart regeneration. To this end, understanding how regenerative responses of the epicardium are regulated in adult zebrafish will lead to new therapeutic targets that underlie the regenerative deficiencies in mammals. To address this, using single-cell RNA-sequencing, we have identified a transient adult epicardial progenitor cell (aEPC) subpopulation within the epicardium after heart injury. Transplantation assays implicate a capacity of aEPCs to give rise to perivascular cells, which are critical for coronary revascularization. Genetic ablation of these aEPCs blocks heart regeneration, suggesting an indispensable role. Pharmacological manipulations and transcriptome analyses yielded candidate genes that underlie the activation of aEPCs. Further, unbiased genome-wide profiling of chromatin accessibility using ATAC-seq revealed putative regulatory elements that exert transcriptional regulation of these genes. We hypothesize that activation of a progenitor cell state in the epicardium underlies successful heart regeneration. To test this hypothesis, we propose to 1) define the cell fates and functions of the aEPCs in adult zebrafish heart regeneration using genetic fate mapping, genetic ablation, and single-cell transplantation approaches; and 2) define the molecular mechanisms underlying aEPC activation through genetic manipulations and analyzing dozens of transgenic lines and mutants. The outcome of this proposal may ultimately inform approaches for activating the epicardial progenitors to enhance the limited regeneration displayed in humans after MI.

NIH Research Projects · FY 2025 · 2021-07

ABSTRACT Right ventricular remodeling leads to serious complications in congenital heart disease. Congenital heart disease (CHD) is the most common birth defect. Due to improved diagnostics and surgery, 1 million patients live in the US with CHD, many of whom develop right ventricular (RV) heart failure. Our understanding of the underlying pathobiology and therapies are very limited, creating a pressing research need. Patients with Tetralogy of Fallot with pulmonary stenosis (ToF/PS), the most common form of cyanotic CHD and the form most available for research, develop adverse RV remodeling, leading to heart failure and arrhythmias. It has been thought that the RV remodeling is a consequence of surgical repair. However, we have recently shown that ToF/PS patients have decreased heart muscle cell (cardiomyocyte) division, indicating the possibility of developing a new mechanistic paradigm of RV heart failure development in CHD. Increased β-adrenergic receptor signaling decreases cardiomyocyte proliferation in ToF/PS. We have taken an innovative research approach, using administration of thymidine labeled with a stable isotope tag (15N-thymidine). Proliferating cells incorporate 15N-thymidine into their DNA, which we visualize with Multi- isotope Imaging Mass Spectrometry (MIMS) analysis of pieces of RV myocardium. By detecting cardiomyocytes labeled with 15N-thymidine, MIMS revealed decreased cardiomyocyte division in ToF/PS. Our mechanistic investigations showed that overactive β-adrenergic receptor signaling inhibits cardiomyocyte division. Our pre-clinical studies in neonatal mice and cardiomyocytes from ToF/PS infants demonstrate that administration of the β-adrenergic receptor blocker propranolol increases cardiomyocyte division. β-blockers have been used in ToF/PS, but this use has been limited to preventing hypercyanotic spells. We propose a randomized, placebo-controlled (1:1), double-blinded, single-center clinical trial of 40 ToF/PS infants to test the mechanistic hypothesis that β-blocker administration in ToF/PS infants increases cardiomyocyte division and decreases RV hypertrophy. The recent success of propranolol administration in infantile hemangiomas and American Academy of Pediatrics guidelines provide the necessary pharmacokinetics and safety experience to support these studies in infants. As primary outcome, we will quantify cardiomyocyte division using our innovative 15N-thymidine labeling approach with MIMS readout. As a secondary outcome, we will characterize changes in RV and cardiomyocyte hypertrophy. This initial single-center trial will provide the foundation for future multi-center randomized controlled trials of propranolol administration in infants with ToF/PS and other types of CHD at risk for RV remodeling, such as hypoplastic left heart syndrome, with the long-term goal of preventing RV failure. The Heart Institute at Children’s Hospital of Pittsburgh is ideal for this research. We have achieved the lowest mortality of infant cardiac surgery and have the research infrastructure to carry out the proposed work.

NIH Research Projects · FY 2025 · 2021-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This T32 grant application seeks support for Training in the Pharmacological Sciences (TIPS) at the Weill Cornell Graduate School (WCGS). This grant will extend support for a highly successful T32 predoctoral training program with current funding slated to end on June 30, 2021. Our TIPS Program is inter-departmental and inter-institutional, comprising 33 outstanding faculty mentors from Weill Cornell Medicine and the Memorial Sloan-Kettering Cancer Center (22 Prof., 4 Assoc. Prof., and 5 Asst. Prof.). We provide a rich research environment for training, including state-of-the-art instrumentation and core facilities, generous space allocation for the TIPS Program, and a continuing commitment to recruiting the brightest and best new faculty. The participating faculty are a cohesive group of world-class investigators with vibrant ongoing research and solid records of training early-stage scientific leaders and scholars. We are dedicated mentors and biomedical researchers with labs that receive >$1.2M in annual average research support. Formal mentorship training is obligate for all of our faculty members and admissions committee members, and we have strict protocols in place for adding and removing training faculty members. A major emphasis of the Program is to share the excitement of discovery with trainees, cultivate the student’s capacity for critical reasoning, and instill in students all necessary skills to fulfill their career aspirations in the many career paths and opportunity our training offers. Notably, 97% of TIPS graduates in the past 10 years have continued in research or a research-related career. Our faculty mentors are highly-collaborative, yet with an array of research interests – providing trainees broad training opportunities in areas that include translational biomedicine, neuropharmacology, cancer biology, cell signaling, metabolism, chemical biology, synthetic chemistry, computational biology and structural biology. The Pharmacology Program uses a rigorous review process for predoctoral applicants that considers overall preparation, motivation and perceived grit, and we enroll 12-16 outstanding trainees annually (90.1% training grant-eligible). During the past 5 years, incoming students had a mean college GPA of 3.58 and 26.2 months of prior research experience. We are proud of the prominent positions held by our graduates, including past/present scientific leaders in government, academia, pharma and biotech - attesting to the impact of our training Program for training the next generation of scientists.

NIH Research Projects · FY 2024 · 2021-07

PROJECT SUMMARY Melanoma is the most lethal of skin cancers, with progression to local invasion and metastasis leading to poor patient outcomes, highlighting the need for better understanding of melanoma progression. During melanoma progression, tumorigenic cells must overcome growth restraints imposed by the microenvironmental keratinocytes. Although much is known of keratinocyte regulatory controls on normal melanocytes, less is known about their interactions in melanoma. Our preliminary data suggests that melanoma induces an epithelial-to- mesenchymal transition (EMT) program in adjacent keratinocytes In Aim 1, I will investigate the role of keratinocyte EMT on melanoma initiation. For this study, I will use the zebrafish as an animal model to study in vivo interactions between keratinocytes and melanoma cells. I will induce spontaneous melanoma formation in transgenic zebrafish lines with GFP labeled keratinocytes and use imaging to confirm morphological changes indicative of EMT in tumor-associated keratinocytes (TAKs). Then, I will assess them for EMT transcription factor and adhesion protein changes. We hypothesize that melanoma-induction of keratinocyte EMT will result in loss of keratinocyte regulation on melanoma proliferation. We will test this hypothesis by knocking out EMT transcription factors in keratinocytes and assess effects on melanoma initiation and proliferation in the zebrafish model. In addition, our preliminary data has also highlighted an upregulation of paracrine signals from TAKs involved in melanoma migration and invasion. In Aim 2, I will determine how keratinocyte-derived secreted factors affect migration and invasion of nascent melanoma. To study migration of melanoma in vivo, I will first optimize existing imaging pipelines in our lab to quantitatively to track cell migration by imaging of the zebrafish skin. I can then assess the effect of knocking out keratinocyte-derived factors such as endothelin, Wnt5A and BDNF using cell-type specific CRISPR-editing to determine their effects on migration. We hypothesize that given the migratory role of these paracrine factors from in vitro data, we will see reduced melanoma migration and invasion into adjacent tissues on migration tracking and histology by knocking-out these factors in keratinocytes. By characterizing the role of tumor-associated keratinocytes in the melanoma microenvironment, this proposal seeks to understand how melanoma modifies its microenvironment to overcome its natural growth restraints and identify new targets to limit melanoma progression.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY/ABSTRACT Christopher Brown MD, PhD is an Instructor of Medicine in the division of infectious diseases at Weill- Cornell Medical College in New York City. His research interests focus on defining the biology of transmission in Mycobacterium tuberculosis. This five-year training grant outlines technical, research and professionalism goals needed to launch an independent academic career. Dr. Brown has extensive research training as an organic chemist and chemical biologist from his doctoral work in the laboratory of Laura L. Kiessling. This proposal expands into the fields of microbiology, metabolomics and regulated gene expression. Utilizing a combination of these disciplines is a powerful method for the molecular interrogation of biological systems. Mtb transmission biology is a field of substantial opportunity for discovery. Development of a transmission blocking therapy would have a profound impact on global health and mortality. The research goal of this proposal is to identify bacterially-encoded determinants that favor successful transmission by imparting survival in air. The PI will expose laboratory and clinical strains of Mtb to air-drying stress and document the effects on metabolism and cellular integrity. Hypotheses regarding the essentiality of metabolic pathways for survival will be tested using CRISPRi (clustered regularly interspersed short palindromic repeat interference) gene silencing.

NIH Research Projects · FY 2025 · 2021-06

ABSTRACT Alzheimer's disease (AD) is the most common form of late-onset dementia. The extents of tau pathology are closely related to memory decline. How pathogenic tau causes cognitive deficits is not clear. While most studies on tau have been focused on direct effects of tau on neurons, compelling human genetic studies linked maladaptive innate immune responses, including microglial responses, to elevated risk of developing late- onset Alzheimer disease. The striking enrichment of innate immune genes as risk alleles for Alzheimer disease supports critical disease-enhancing role of maladaptive microglia in tau-mediated cognitive deficits. Identifying how maladaptive microglial enhances tau toxicity could lead to new therapeutic strategies. In our preliminary studies, we found that tauopathy mice exhibit hyperactive Cyclic GMP-AMP synthase (cGAS)- Stimulator of interferon genes (STING) signaling. As a major sensor of cytosolic DNA, cGAS-STING pathway mobilizes antiviral responses via activation of interferon regulatory factors (IRFs) and expression of cytokine and type I interferon genes. The hyperactive cGAS pathways contributes to tau toxicity since genetic reduction of cGAS protected against tau-mediated spatial learning and memory deficits in a tauopathy mouse model of Alzheimer disease. In addition, the protective effects were associated with reduced interferon-enriched microglial subpopulations and reprogramming of disease-associated microglial states as identified using single nuclei RNA-seq. We hypothesize that microglial cGAS-STING hyperactivation mediates the maladaptive disease-enhancing microglial response in tauopathy. To test this hypothesis, In Aim 1, we will first determine how cGAS activation in microglia enhances tau toxicity (1a). Using a combination of single nuclei RNA-seq, pathological and functional analyses, we will investigate if toxicity from cGAS hyperactivation in microglia or bone marrow-derived monocytes (1b, 1c). In Aim 2, we will dissect if the toxic effects of cGAS activation in tauopathy are mediated by STING-dependent or -independent mechanisms, we will determine if loss of STING phenocopies the effects of cGAS inactivation on tau toxicity and transcriptomic changes (2a). We will then determine STING-independent mechanisms of cGAS activation by assessing how cGAS inactivation affects tau toxicity on Sting null background (2b). The significant protective effects of partial loss of cGAS supports that partial inhibition with pharmacological inhibitors of cGAS could be beneficial for tauopathy. We showed that a specific cGAS inhibitor, TDI6570, is brain permeable and effectively inhibit expression of type 1 interferon target genes in tauopathy mice. We will then optimize the dosing using formulated chow based on PK/PD, and evaluate the beneficial effects of the cGAS inhibitor before or after the onset of cognitive deficit in tauopathy mice in Aim 3. Completion of the proposed study will identify novel disease-enhancing properties of innate immune responses in AD, and provide new therapeutic direction for pharmacological intervention.

NIH Research Projects · FY 2025 · 2021-06

Abstract Immunoglobulin A (IgA) is prominently secreted at mucosal surfaces and coats a fraction of the intestinal bacterial microbiota. In health and disease, secretory IgA (sIgA) binding influences intestinal immunity and homeostasis by crosslinking microbiota in the lumen to prevent encroachment on the intestinal epithelium, shuttling bound microbes to secondary lymphoid tissues, and directly modulating microbial metabolic activity. Aside from the “natural” polyreactive IgA detectable in germ-free mice, sIgA is predominantly gut colonization dependent. The identification of immunogenic commensal bacteria and their specific IgA epitopes have further elucidated our understanding of the mechanisms governing gastrointestinal balance and how dysbiosis can drive intestinal pathologies. Meanwhile, the potential involvement of the fungal component of the gut microbiota (mycobiota) in these processes is largely unknown. Only recently have intestinal fungi been recognized as a factor contributing to events associated with inflammatory disease or response to therapy prompting multiple questions regarding the development of antifungal mucosal antibody responses, their specificity, and mechanisms of induction in the gut. In recent work, we have shown that polymorphisms in the Dectin-1 gene (CLEC7A) or the fractalkine receptor gene CX3CR1 are associated with defects in antifungal immunity in Inflammatory Bowel Disease (IBD) patients, and notably the latter leading to gut fungal overgrowth and substantial decrease of antifungal antibodies. In preliminary studies we unexpectedly identified a broad range of fungal organisms that were targeted by sIgA antibodies. Hyphal formation is a primary mechanism used by many dimorphic fungi to invade and survive within their hosts. Notably we found that mycobiota aggravated intestinal damage and inflammation is dependent upon hyphae-produced virulence factors that are targets of sIgA. These preliminary data support the overall hypothesis that antifungal sIgA antibody responses are naturally induced by specific gut mycobiota species and act against fungi-produced factors to play a key role in mucosal immunity by averting direct contact of fungi with the intestinal epithelium to prevent intestinal barrier damage and related gut inflammation. We will investigate this hypothesis both in vitro and in in vivo models, aided by deep sequencing and computational platforms, and genetically modified fungal strains. We will determine IgA-reactive gut mycobiota and fungal morphotypes involved in the induction of antifungal sIgA antibodies and will make use of several model systems to define the functional role of antifungal sIgA in gut.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY Current HIV antiretroviral treatment successfully controls viral replication and has transformed HIV-infection from a fatal illness to a manageable chronic condition. However, despite suppression of viral replication during treatment, studies have shown that pools of latent viral reservoirs remain detectable, which fuel viral rebound when antiviral suppression treatment is interrupted. These viral reservoirs are established almost immediately upon infection when HIV irreversibly integrates its viral genome into human DNA. Viral reservoirs are extremely durable, not susceptible to therapeutic effects of currently available antiretroviral agents, and have been refractory to recent experimental treatment approaches. HIV infection is also characterized by a high level of intrahost genotypic diversity of viral quasispecies. In addition to genetic diversity associated base substitution mutations, pools of viral DNA genomes recovered from chronically-infected patients under prolonged suppressive therapy often contain high frequencies of genome-truncated and/or hypermutated, non-replication- competent viral DNA genomes. Only a small fraction of proviral genomes in these patients are genome-intact and may lead to productive viral replication and virologic rebound in the absence of treatment. Furthermore, HIV-infected cells infected with both genome-intact and genome-defective proviruses have been shown to clonally expand, serving as a mechanism of HIV persistence. However, our current understanding of HIV reservoirs has been derived almost exclusively from studies on a strain called subtype B HIV-1, the predominate viral subtype affecting first-world nations but only makes up 10% of the global epidemic. In contrast, non-B HIV- 1 subtypes predominate regions such as sub-Saharan Africa where disease burden is the highest globally. Questions remain on whether the remaining 90% of infections by other HIV-1 subtypes differ in reservoir sizes and compositions. To address this question, we will leverage an existing biobank of a previously NIH-funded Ugandan HIV cohort (UARTO), which houses 12360 blood cell samples collected longitudinally over ten years from 500 predominantly subtype A1 and D HIV-1-infected individuals. We will use three cutting-edge technologies (1) FLIP-seq to obtain near-full-length HIV-1 DNA genomes profiles, (2) MIP-seq to co-capture HIV- 1 integration sites and viral genome, and (3) and the Intact Proviral DNA Assay (IPDA) to longitudinally measure the decay/expansion rate of the reservoir. All three technologies allow us to focus on the rare intact viral DNA genomes that is the target for HIV cure strategies. Across subtypes, we will compare reservoir characteristics including absolute genome-intact reservoir sizes, extent of clonal expansion, integration site profiles, viral promoter genotypes, and longitudinal decay/expansion dynamics. We will further investigate demographic, clinical and host factors associated with genome-intact viruses. Overall, we aim identify differences, or the lack of differences, between HIV-1 subtype reservoirs to inform HIV cure research effort on whether a cure strategy should be subtype-specific.

NIH Research Projects · FY 2025 · 2021-06

Project Summary Coronary artery disease (CAD) is a leading cause of morbidity and mortality: revascularization via coronary artery bypass grafting (CABG) is a common therapy for CAD, which is performed via either single arterial graft (SAG) or multiple arterial grafting (MAG) strategies. Whereas arterial grafts have better long-term patency than vein grafts, data indicates that patients receiving MAG may experience higher perioperative risk. Cardiac function is known to predict prognosis after CABG, but the impact of different revascularization approaches on cardiac performance and consequences on clinical outcomes are incompletely understood. In this re- submission K23 proposal, I will build on my background in cardiothoracic anesthesia and advanced echo imaging to elucidate the effect of the two different revascularization strategies (SAG vs. MAG) on cardiac function immediately after CABG and subsequent clinical outcomes, in order to provide a framework for perioperative management of CABG patients. In Aim 1, I will examine the impact of CABG revascularization strategy on the change in cardiac function defined by echo strain. I will identify if MAG will result in lower cardiac performance as quantified by transesophageal echocardiography from baseline to after the operation. I will also evaluate whether the change echo strain will be associated with medications need to support hemodynamics. In Aim 2, I will determine the mechanism by which echo strain decreases after CABG. Myocardial perfusion on contrast-enhanced echo and flowmeter will be utilized to test the association with cardiac functional change (echo strain). I will also evaluate if MAG will result in lower myocardial perfusion than will SAG-based revascularization. In Aim 3, I will determine whether the decrease in echo strain predicts clinical outcomes at 6 months better than conventional imaging indices. By studying the link between revascularization techniques, cardiac function and physiology, I will lay the groundwork for a research career translating new insights regarding underlying mechanisms of cardiac dysfunction into meaningful anesthetic interventions that improve clinical outcomes. Under my strong mentorship team of Drs. Devereux (extensive expertise in cardiovascular imaging trials and cardiac outcomes research), Gaudino (internationally renowned cardiac surgeon with a focus on multi-arterial grafting and designing randomized clinical trials and m-PI of the funded parent trial ROMA), and Weinsaft (an expert in quantitative cardiac imaging and translational research), I will be able to enhance my clinical research skills while obtaining new technical skills advanced echocardiography and translational research that will allow me to become an independent investigator. In addition, new skills obtained from my K23 will directly guide the design of a planned R01 to (a) to assess LV response to vasoactive medications designed to improve arterial graft flow in CABG patients and (b) test vasodilator therapy to improve clinical outcomes in patients with MAG.

NIH Research Projects · FY 2025 · 2021-06

PROJECT ABSTRACT Inflammation in the central nervous system (CNS) is causally associated with the pathogenesis and progression of multiple demyelinating and neurodegenerative diseases, including Multiple Sclerosis (MS), Parkinson's Disease, Alzheimer's Disease, Amyotrophic Lateral Sclerosis, and general age-associated cognitive decline Therefore, there is an urgent need to develop novel approaches to prevent, limit or reverse neuroinflammation. Basic, translation and clinical studies indicate that T cells can be major drivers of inflammation in the CNS, but the mechanisms promoting or inhibiting these responses remain poorly defined. The fundamental focus of this research proposal is to: (i) interrogate a novel pathway by which the innate immune system controls T cell responses in the CNS, (ii) understand the regulation of these cellular interactions, and (iii) define whether it is possible to harness this or related pathways for therapeutic benefit in neuroinflammation. We will employ innovative approaches and develop new tools to address these fundamental gaps in knowledge, and where possible, translate our findings from mice into clinically defined patient samples. Results from these studies will advance our understanding of the pathways that promote or inhibit pro-inflammatory T cell responses in the CNS, and could provoke the next generation of novel preventative, therapeutic or curative treatment strategies for demyelinating and neurodegenerative diseases.