Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Stroke affects nearly 800,000 Americans each year. Assigning the risk of a future stroke is feasible based on established risk factors, such as a carotid stenosis, but determining the timing of an ischemic event is difficult due to the unpredictability of the timing and final location of a thromboembolism in the brain. In many cases, it is the occurrence of stroke that leads to assessment of risk factors which then necessitates monitoring or an intervention such as a stenting. Surgical procedures themselves have risk of causing stroke from release of emboli, and during recovery there is a risk of a stroke related to antithrombotic drug interruption and transient hypercoagulability. Methods to detect emboli in at-risk patients or to monitor for emboli during or after surgery therefore have clinical utility. Transcranial Doppler (TCD) ultrasound (US) is commonly used to assess blood flow to the brain and to monitor for emboli. However, current TCD is cumbersome and does not allow long term, automated detection for emboli outside the clinical setting. We will address this gap in technology through the development of a compact, portable TCD US device that will improve the ability to monitor for emboli in a clinical setting, but more importantly, to monitor at risk patients in an ambulatory nonclinical setting. We will build a novel TCD US array and FPGA-based control module capable of collecting data over long periods of time and automate the detection of emboli using deep learning (DL). As with current TCD, the device will insonate through the transtemporal acoustic windows. The 2D array will permit beam steering to automatically detect the middle cerebral artery (MCA). Once the MCA is located, sophisticated signal processing and DL algorithms will be employed to flag passing emboli. Aim 1 will focus on the design, fabrication and testing of the TCD array. The novel 1.5 MHz TCD array is based on a sparse 16 element design. The device will be tested using a flow phantom to collect in vitro acoustic signatures of simulated emboli. After establishing acoustic safety limits, a limited study will be undertaken with healthy subjects. In Aim 2, we will first implement and validate DL architectures using the Aim 1 in vitro data. We will use the TCD array to collect in vivo emboli data during right-to-left shunt bubble tests and then retrain the DL model and test DL emboli detection in a prospective clinical analysis. For Aim 3, we will build an FPGA-based TCD US array control module and, once the DL methods are optimized in Aim 2, embed them on the FPGA module for autonomous emboli detection. Finally, in Aim 4, we will test our system during cardiac surgery where quasi-random release of emboli (gaseous and/or particulate) is highly correlated with standard surgical steps. Our autonomous TCD system will open a path to wearable, long term monitoring for emboli in at risk patients.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Immunotherapies, mainly immune checkpoint blockade (ICB; e.g., aPD-1 and aCTLA-4), cause immune-related adverse events (irAEs) in 20-30% of cancer patients, with skin and gastrointestinal toxicities being most common. High-dose corticosteroids are typically the first line of treatment for irAEs but timing, resistance and side effects complicate their use and ICB treatment. Critically, the mechanisms driving irAEs remain unclear, limiting the design of targeted therapies that reduce irAEs while preserving anti-tumor activity. In this proposal, we use skin irAEs as a model system to dissect these mechanisms. Our preliminary data showed that neutrophils play a role in both tumor eradication and skin irAEs in T cell therapy, with VISTA+ neutrophils as the predominant population in irAE-affected skin. VISTA targeting reduced irAEs without affecting anti-tumor immunity. Separately, we found that, in non-tumor models, ICB can unleash inflammatory responses from commensal-specific T cells, promoting neutrophil infiltration and irAEs. Notably, metabolic interventions that lower blood glucose (the main energy source for T cells and neutrophils), including caloric restriction (CR) and a pharmacologic CR-mimic, mitigate these irAEs. However, these strategies require further validation in tumor models following ICB for mitigation of irAEs without losing anti-tumor effects. Additionally, we showed that ICB induces loss of regulatory T cells (Tregs), which may disrupt tolerance to skin commensals and allow commensal-specific T cells to cross-react with tumor antigens and drive irAEs. In this scenario, we hypothesize that immunotherapy induces irAEs by activating commensal-specific T cell and pathogenic neutrophil responses, which can be modulated with targeted or metabolic interventions to prevent toxicities while preserving anti-tumor immunity. To test our hypotheses, we propose the following: In Aim 1, we will define the role of VISTA+ neutrophils in irAEs during T cell therapy using VISTA antibodies and knockout (KO) models, complemented with comprehensive immuno-phenotyping analyses. We will also investigate how VISTA+ neutrophils cause irAEs in ex vivo assays and assess VISTA’s value as a skin irAE biomarker using patient samples. In Aim 2, we will assess the therapeutic potential of blood glucose-lowering nutritional (CR) and pharmacological (tirzepatide) interventions in limiting commensal-specific T cell and neutrophil function to reduce irAEs while maintaining anti-tumor responses. We will also directly assess the role of glycolysis in fueling commensal-specific T cells using T cell- specific glycolysis KO and CRISPR models. In Aim 3, we will investigate whether and how ICB disrupts skin commensal tolerance and induces irAEs following ICB and interrogate commensal/tumor T cell cross-reactivity as one of the underlying causes of irAEs via TCR-seq and skin commensal ablation. Our long-term goal is to develop effective therapies that mitigate irAEs while preserving anti-tumor efficacy for improving precision and safety of immunotherapies. With VISTA antibodies already in trials, simple dietary interventions, and FDA-approved tirzepatide, our findings are poised for rapid translation and clinical use.

NIH Research Projects · FY 2026 · 2026-06

Cervical cancer mortality is preventable when detected by screening and treated at an early or precursor stage. Despite similar rates of cervical cancer screening, women with limited financial means or education, or low health literacy suffer from disproportionately higher rates of cervical cancer incidence and mortality compared to average women. A major influence on these outcomes is lack of understanding of the importance of follow-up after abnormal screening. Effective and scalable educational interventions are currently not widely implemented. An intervention emphasizing the importance of colposcopy and follow-up after abnormal cervical cancer screening that is easily understood by women in high-risk groups is a critical strategy to reduce cancer- related mortality. We propose a hybrid type 1a implementation-effectiveness trial to test the hypothesis that a pilot tested educational video intervention called COLPO (Clear communication, Optimize Follow up, Leverage Education, Promote prevention, Outreach to increase awareness) reduces loss to follow-up after colposcopy in high-risk women. COLPO uses a novel approach to education called the Patient Activated Learning System (PALS), which has been proven to increase knowledge uptake and receptivity to medical advice in low-income populations. On detection of an abnormal cervical cancer screening, women are sent SMS text messages with links to PALS videos explaining information about the human papilloma virus and the importance of follow-up. In Aim 1 we will engage stakeholders to refine the intervention protocol guided by the CFIR framework and our Community Advisory Board (CAB). We will refine the videos based on feedback of participants in our pilot study. In Aim 2, we will conduct the trial by engaging 700 women with abnormal Pap screening results who will be randomly assigned to receive either the COLPO educational intervention or standard follow-up care. The women will be patients at one of our 4 clinics in New York City. The primary effectiveness outcome is timely colposcopy follow-up appointments within 3 months. An exploratory effectiveness outcome is guideline concordant care within 1 year, measured via EMR surveillance. In Aim 3 we will assess implementation of the intervention guided by the RE-AIM (Reach Effectiveness Adoption Implementation Maintenance) framework by collecting survey data on patient satisfaction, anxiety, confidence, and knowledge, as well as qualitative data related to implementation. We will also embed the COLPO intervention into our EMR as a model for scalability. We will work with the CAB to disseminate the findings in the community. Our multidisciplinary team includes experts in Gyn/Oncology, the PALS, qualitative research, implementation trials, and EMR implementation, enhancing the likelihood of success of this project. If COLPO proves effective, the intervention will be scaled across our 11-hospital health system to increase colposcopy and follow-up among the Meyer Cancer Center catchment area serving more than 6 million patients, with the goal of reducing cervical cancer incidence and mortality for all populations.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT - OVERALL Efforts to establish control of the tuberculosis (TB) pandemic face historically unprecedented challenges. In the wake of COVID-19-induced disruptions in TB healthcare-related services, TB has re-emerged as the leading cause of deaths due to an infectious disease with 2 consecutive years of the first increases in TB mortality seen in over a decade. Lacking more effective and scalable transmission blocking interventions, control of the TB pandemic will continue to rest on the clinical efficacy of chemotherapy. Unfortunately, while still therapeutically effective against 95% of newly diagnosed cases, existing chemotherapies have proven functionally far less effective due to their toxicities and months-long duration. This Program Project proposes to lay a basic-science foundation for developing treatment shortening drugs and regimens. Such studies are motivated by the empirical discovery of the treatment shortening properties of rifampin and pyrazinamide, two frontline agents whose addition enabled a nearly 3-fold reduction in treatment duration, and more recent evidence implicating phenotypically drug-resistant bacterial subpopulations as a biological determinant of treatment duration. This application brings a synergistic combination of investigators and disciplines together for a collaborative attack that mobilizes new concepts, technologies and model systems to discover and test new treatment shortening targets and strategies. Project 1 focuses on mechanisms that allow these populations to persist in patients undergoing chemotherapy. Project 2 focuses on reverse translating the treatment shortening activities of rifampin and pyrazinamide into in vitro biomarkers, mechanisms and targets. Project 3 will use innovative CRISPRi-based genetic-genetic interaction mapping to identify highly efficacious Mtb drug-target combinations. Project 4 exploits genetic approaches to define the Mtb gene products whose inactivation has the greatest potential to shorten TB chemotherapy. Core A ensures the efficient flow of information and material among these projects. Core B will combine technical advances in single-cell measurement with data integration approaches to enable single cell studies of how different subpopulations respond to treatment and/or gene inactivation. Core C will provide access to a novel paucibacillary and a drug treatment mouse model of TB relapse.

NIH Research Projects · FY 2026 · 2026-05

Clinical outcomes of Mycobacterium tuberculosis (Mtb) infection are highly variable and shaped by complex host-pathogen-environment interactions. The goal of this program is to improve understanding how Mtb is continuing to evolve essential biological pathways to improve its fitness within the human population. We focus on genes and pathways under positive selection in clinical isolates—key determinants shaped by immune and antibiotic pressures—aiming to clarify their roles in clinical outcomes. Project 1 investigates the functional consequences for TB pathogenesis and antibiotic susceptibility of clinically selected mutations in metabolic enzymes. Project 2 investigates how Mtb uses cyclic AMP (cAMP) signaling to adapt to the host environment and resist chemotherapy. Project 3 dissects how clinically selected mutations in the type VII ESX-1 secretion system affect virulence, antigenicity and drug sensitivity. Project 4 identifies post-transcriptional mechanisms that buffer the fitness costs of evolving mutations. Core A coordinates the activities of the four projects and two scientific cores, and manages progress to goals, resources, and communications. Core B determines the impact of clinically selected mutations on protein activity, stability and structure. Core C performs phylogenomic analysis of a diverse, global collection of Mtb genomic data, tabulates data on genetic variants and signals of selection, and identifies epistatic interactions with them. The proposed research integrates cutting-edge genomic, phenotypic and biochemical analyses to decipher drivers of selection and adaptation, offering a novel framework to explain host-pathogen interactions that dictate disease progression.

NIH Research Projects · FY 2026 · 2026-05

Project Summary / Abstract Melanoma is a striking example of a cancer where a subset of patients are effectively treated, while others are resistant to the same therapies or develop resistance after an initial positive response. This resistance underscores the unmet need to develop new therapies and new strategies to potentiate existing therapies. Inter- cellular interactions with the melanoma microenvironment control tumor initiation and progression and are widely considered critical to the efficacy of chemotherapies and immunotherapies. Axons, the cable-like projections that connect neurons with their target cells, are a newly discovered component of many tumor microenvironments, including amongst melanomas. However, how the nervous system signals within melanomas and the cells within the melanoma microenvironment that they target are largely unknown, critical prerequisites to determining whether axons are viable therapeutic targets. Our project is focused on defining how axons of the sympathetic nervous system inhibit melanoma growth, focused on interactions between these axons and immune effectors. Sympathetic axons release the neurotransmitter norepinephrine that activates adrenergic receptors on target cells. Our first goal is to define the cell types in the melanoma microenvironment that express these receptors and, through genetic analysis, their functional requirement for these receptors for axonal inhibition of melanoma growth. Here, our specific focus is on adrenergic receptors in myeloid derived cells including macrophages as part of our broader hypothesis that macrophages are critical effectors of sympathetic nerve-derived inhibitory signals (Aim 1). Next, we will focus on the anti-tumor immune response that sympathetic axons initiate. We will determine the degree to which intra-tumoral axons are susceptible to and protected from inflammatory damage that may limit their capacity to effectively signal to their targets as tumors grow (Aim 2). These experiments are aided by our ability to perform whole tumor imaging and to reconstruct and quantify the tumor-wide axonal innervation patterns. Finally, we will test whether stimulation of sympathetic axons or the adrenergic receptors that they activate reduces melanoma progression either alone or in combination with established anti-melanoma therapies such as immune checkpoint blockade (Aim 3). Insights from our studies are expected to inform new anti-melanoma strategies that harness the immune-regulatory function of intra-tumoral sympathetic axons.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Adoptive cell therapies (ACT) with chimeric antigen receptors (CAR) hold promise to treat cancer, yet several challenges must be overcome to achieve this goal. Clinical evidence attributes treatment failure and resistance to the lack of functional T-cell persistence, antigen escape, and a suppressive tumor microenvironment replete with inhibitory but not co-stimulatory molecules. Investigating response and relapse mechanisms and effectively targeting tumor resistance mediators are urgently needed to improve current ACT. Our analysis of clinical CAR-T products revealed intrinsic PD-L1 (int-PD-L1) expression on the T cells and trogocytic acquisition of tumor-derived PD-L1 (extrinsic; ext-PD-L1). However, the functional role of PD-L1 on CAR-T remains unknown. We hypothesize that both int-PD-L1 and ext-PD-L1 limit CAR-T activity, and that rewiring PD-L1 and CD80 signaling with synthetic receptors will boost antitumor efficacy across ACT platforms. Aim 1 will determine how int-PD-L1 regulates CAR-T under optimal and suboptimal antigen stimulation; Aim 2 will define the impact of trogocytic ext-PD-L1 transfer on CAR-T function using in vitro assays and in vivo leukemia and melanoma models. To enhance T-cell persistence and function, we developed 80BB, a synthetic dual-costimulatory receptor fusing the CD80 ectodomain to the 4-1BB endodomain. 80BB delivers CD28 and 4- 1BB signals and functions as a CTLA-4 switch receptor, driving tumor control across diverse ACT platforms. Although CD80 interaction with CD28 and CTLA-4 underlies these benefits, CD80 also binds PD-L1, yet 80BB did not protect CAR-T from PD-L1/PD-1-mediated inhibition. We therefore engineered a novel 80BB-based platform that retains CD80 interaction with CD28 and CTLA-4 while redirecting PD-L1 engagement, to overcome PD-L1-mediated suppression. Aim 3 will determine mechanisms of action and therapeutic benefit of this novel synthetic costimulatory receptor in B-cell malignancy and solid-tumor models. Our multidisciplinary approach combines basic immunology with translational immunoengineering to overcome ACT barriers. We aim to advance next-generation engineered T-cells and expand the scope of ACT, improving the survival and quality of life of cancer patients.

NIH Research Projects · FY 2026 · 2026-05

Overall Abstract This HIVRAD Program Project application is made in response to PAR-24-037. It contains two Research Projects, one Scientific Core and an Administrative Core, under the direction of Principal Investigator, John P. Moore, PhD (Weill Cornell Medicine; WCM) and co-Principal Investigator, Ian A. Wilson, PhD (The Scripps Research Institute; TSRI). The third performance site is the Academic University Medical Centers (AUMC), Amsterdam. The goal of the project is to further develop stable, soluble, cleaved trimeric mimics of the native Env spike (SOSIP trimers) as germline-targeting immunogens for the development of vaccines aimed at inducing broadly neutralizing antibodies (bNAbs), and as antigens for structural studies that improve trimer design. Our central hypothesis is that proteolytically cleaved, soluble, trimeric forms of HIV-1 Env (SOSIP trimers) are appropriate structural antigens for high resolution x-ray crystallography and electron microscopy studies, and suitable immunogens for vaccine research aimed at the induction of bNAbs. Our intent is to use structure-based information to help develop an effective, prophylactic HIV-1 vaccine (or component of a more complex vaccine) that is based on the concept of inducing sufficient titers of bNAbs. SOSIP trimers have the desired properties and can be produced efficiently, including in the amounts and qualities required for human clinical trials. We will focus on the use of appropriately modified SOSIP trimers that are able to initiate germline-bNAb lineages that can then be shaped and polished into mature antibodies with the properties possessed by bNAbs. The same HIVRAD program team has made excellent progress during the past 5 years, and now seeks support to continue to work together. Its research plan involves the following sub-components. Project 1: Rogier W. Sanders and PJ Klasse (with Marit van Gils): Design of germline-targeting trimers Project 2: Ian A. Wilson (with Andrew B. Ward): Structure guided design and refinement of trimers Core B: John P. Moore: Production of SOSIP trimers Core A: John P. Moore: Administrative support Marit van Gils (AUMC) will co-Lead Project 1. Andrew B. Ward (TSRI) will co-Lead Project 2. As well as the integral components of the program team, we propose to maintain and expand an extensive network of research collaborations, and we will continue to provide SOSIP trimers and related reagents to the many scientists who request our support.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY Anti-retroviral therapy suppresses HIV replication but fails to completely eradicate the virus, which persists in a reservoir of infected CD4+ T cells that resist immune-mediated clearance. Immune evasion by reservoir cells has canonically been attributed to viral latency, which silences HIV gene expression and thereby precludes the recognition of infected cells by HIV-specific cytotoxic T lymphocytes (CTLs). It is now clear, however, that some reservoir cells continue to express HIV antigens, raising the question of how they resist killing by CTLs. To identify these latency-independent mechanisms of resistance, we have employed molecular, cellular, and biophysical approaches to analyze HIV-infected T cells that survive extended coculture with cognate CTLs. Our results indicate that survivor cells express lower levels of cytoskeletal proteins and cell surface adhesion molecules, accompanied by reduced cortical stiffness and plasma membrane tension. These architectural and mechanical phenotypes are intriguing because CTLs kill their targets using an adhesive and physically active immune synapse, and recent studies indicate that perturbing the biophysical properties of target cells can protect them from CTL-mediated destruction. Accordingly, we hypothesize that a subset of HIV-infected cells elude the cytotoxic immune system by becoming soft and slippery. Our proposed studies, which are divided into two Specific Aims, will investigate the molecular mechanisms underlying these resistance pathways with the end goal of identifying novel strategies for targeting the HIV reservoir. Aim 1 will combine biophysical and immunological assays with in vivo experiments to determine the basis for cell softening in resistant cells. We are particularly interested in the interplay between HIV Nef, a viral protein known to induce actin remodeling, and the host cell cytoskeleton. Aim 2 will focus on the molecular foundation(s) of the adhesion phenotype. Our preliminary studies suggest a role for the histone methyltransferase EZH2, which has been shown to mediate the downregulation of adhesion molecules in the context of B cell lymphoma. This proposal is anchored by the conceptually innovative idea that HIV exploits biophysical vulnerabilities to achieve immune evasion, and it will be carried out using technically innovative approaches and reagents, including single cell biophysical measurements as well as first-of-their-kind authentic reservoir CD4+ T cells clones isolated directly from people with HIV. We will also establish avenues for clinical translation based on small molecule modulators of the cytoskeleton and Tazemetostat, a clinically approved EZH2 inhibitor. These efforts will incorporate samples and data from an ongoing Tazemetostat clinical trial, enabling us to evaluate the effects of this drug in actual patients. The successful realization of our Specific Aims has the potential to substantially broaden the of study HIV resistance mechanisms, paving the way for a different kind of cure research. As such, this work is highly relevant to the NIH mission in that it will contribute to the advancement of knowledge that could improve human health.

NIH Research Projects · FY 2026 · 2026-05

The translocation of polar lipids from one side of a membrane bilayer to the other is critically important in physiology. This process, termed lipid flip-flop, is variously required for biogenesis of mitochondria, growth of the endoplasmic reticulum (ER), all forms of protein glycosylation, and synthesis of glycolipid blood group antigens. It is also needed to expose the signaling lipid phosphatidylserine (PS) at the cell surface in response to physiological triggers – PS promotes blood clotting by activated platelets, marks apoptotic cells for clearance by macrophages and induces cell-cell fusion needed to produce myotubes and osteclasts. Specific proteins – termed scramblases – catalyze lipid flip-flop by acting as lipid channels. These proteins have attracted considerable recent interest, resulting in new information and new concepts concerning intracellular lipid transport and homeostasis, yet many fundamental mechanistic and biological questions remain. The overall goals of our research are to investigate these questions broadly, by discovering the molecular identity of scramblases in key processes where their activity is implicated but not yet been assigned to a specific protein(s), understanding the molecular mechanisms by which these novel lipid channels work, evaluating their specific contributions to physiological processes, and understanding how their activity is regulated. To achieve these goals, we focus here on two understudied research areas which exemplify key knowledge gaps. We recently discovered phospholipid scramblases in the outer mitochondrial membrane. These proteins likely provide the means by which polar lipids enter mitochondria to support the growth and function of the organelle. We will use a multidisciplinary suite of methods to understand how these proteins work and to define their specific contributions to mitochondrial biogenesis. Second, we are interested in the molecular identity of glycolipid scramblases that are implicated in all forms of protein glycosylation in the ER. These scramblases have eluded discovery, and their identification is a major goal for the field. We propose a multi-pronged experimental approach to identify these proteins and validate their function using biochemistry and genetics. Mitochondrial dysfunction is a common source of inborn errors of metabolism, and is associated with cancer, cardiovascular and neurodegenerative diseases, as well as with lipid syndromes such as Barth. Protein glycosylation is essential for life, and specific defects in glycosylation pathways can lead to a number of diseases including muscular dystrophy. In addition to advancing the fields of mitochondrial cell biology and glycobiology and providing unifying biophysical insights into a fundamental membrane transport process, the research proposed here and the overall program that we have conceived will contribute to an understanding of the etiology and presentation of many diseases.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Prostate cancer arises as an androgen driven disease, and androgen receptor (AR)-targeted therapies are the mainstay of treatment for men with advanced disease. One mechanism of resistance is ahistologic transformation from an AR-driven prostate adenocarcinoma to an AR-independent small cell neuroendocrine carcinoma, often referred to as neuroendocrine prostate cancer (NEPC). NEPC is clinically aggressive, frequently metastasizes to visceral organs, and carries a poor prognosis. A thorough molecular understanding of NEPC progression is needed for the development of strategies to treat, prevent, or reverse the development of this lethal disease. Although NEPC tumors arise clonally from prostate adenocarcinoma, there is significant epigenetic and transcriptomic dysregulation that occurs during the lineage plasticity process. Mechanistically, we still do not know how these alterations arise and how best to leverage these alterations as a therapeutic opportunity. Based on published reports and on our preliminary data, E2F1 is over-expressed in the majority of NEPC cases and in a subset of CRPC cases and is associated with a poorer prognosis compared to CRPC with low to no E2F1 expression. However, little is known about the role of E2F1 in the progression from CRPC to NEPC. Our preliminary and published data from patient tumors and in vivo, in vitro and ex vivo models (patient- derived organoids) suggest that E2F1 drives a reprogramming of chromatin accessibility which in turn, results in a NEPC-associated change in gene expression and that this is potentially mediated through a physically interaction with specific NEPC-associated co-factors and transcription factors. Our long-term goal is to develop new biomarker-driven therapeutic strategies for treating patients with advanced prostate cancer and untreatable NEPC. The objective here is to identify the key molecular events and mechanisms underlying lineage plasticity in prostate cancer. This would allow for early therapeutic intervention and improve patient outcome. Our over- arching hypothesis, which is based on our published and preliminary data, is that specific molecular alterations (e.g. RB1 loss and E2F1 induction) in prostate cancer cells drive the progression of CRPC tumors towards NEPC resulting in changes to chromatin accessibility and interactions with specific pro-stem cell- and neural lineage- associated transcription factors to drive a NEPC-associated change in gene expression. To address this hypothesis, we will define essential E2F1-transcriptional complex proteins that mediate the gene expression program driving and maintaining NEPC-progression (Aim 1) and determine if E2F1 is essential in mediating the gene expression program associated with the transition from CRPC towards NEPC (Aim 2). Successful completion of these Aims will provide unique insights into NEPC development, identify potential targetable mediators of lineage plasticity, and a timely and unique opportunity for the early detection of patients with E2F1- expressing CRPC that are evolving towards NEPC that may not respond to standard-of-care anti-AR therapy.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Mycobacterium tuberculosis (Mtb) is a leading cause of death from infection. Curative drug regimens are long, plagued by resistance and often toxic. In addition to using antimycobacterial drugs, adjunctive host directed therapy (HDT) aims to modulate host immune and inflammatory responses to reduce tissue damage, treatment duration and relapse rate. Here we will explore a unique way to combine pathogen-directed and host-directed therapy by exploiting the contrasting ways that Mtb and macrophages, the predominant niche for Mtb, handle cholesterol, Mtb's preferred carbon source. Mtb-infected macrophages form droplets filled with cholesterol esters and triacylglycerides that sustain the bacterium and induce an antibiotic-tolerant state. However, Mtb's degradation of cholesterol generates a toxic intermediate, propionyl-CoA. Mtb avoids propionyl-CoA accumulation by assimilating propionyl-CoA into virulence lipids (PDIMs and sulfolipids). Inhibition of the essential Mtb enzyme phosphopantetheinyl transferase (PptT) blocks synthesis of PDIMS and sulfolipids by inhibiting charging of the acyl carrier proteins required for their synthesis. We found that anti-mycobacterial agents that inhibit PptT kill Mtb in association with propionyl-CoA accumulation and CoA depletion. Moreover, we found that propionyl-CoA directly inhibits Mtb CoaBC in vitro, an essential enzyme for CoA biosynthesis. This likely explains why propionyl-CoA accumulation is toxic. Here, we will harness the PptT–PDIM–propionyl-CoA– CoaBC axis to kill intracellular Mtb by the cumulative effects of inhibition of Mtb's PptT and the host macrophage's sterol-O-acyltransferase (SOAT), an enzyme crucial for conversion of free cholesterol to its esters. Inhibition of SOAT will lead to accumulation of free cholesterol rather than its storage as esters. Mtb's metabolism of free cholesterol will exacerbate killing of Mtb by PptT inhibitors and the consequent prevention of propionyl- CoA incorporation into lipids, leading to the blockade of CoA synthesis. Inhibition of both SOAT isoforms (1 and 2) can prevent lipid droplet formation, and chemical inhibition of SOAT and its knockdown by RNA interference have been shown to restrict intracellular bacterial growth. However, chemical inhibition of SOATs 1 + 2 has resulted in toxicity due to lack of isoform specificity. Herein, we will use a new series of SOAT1 inhibitors that are selective for the SOAT1 isoform, which predominates in macrophages. We will test the efficacy of SOAT1 inhibition on intracellular growth of Mtb in conditions of lipid droplet induction that lead to different ratios of cholesterol esters and triacylglycerides. We will treat macrophages with SOAT1 inhibitor and lipid droplet inducer, infect them with Mtb, treat with our highly potent and selective PptT inhibitors and monitor killing of Mtb.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Parkinson's Disease (PD) is the second most prevalent neurodegenerative disorder all over the world. It is estimated that PD affects 2-3% of people older than 65 years. PD is associated with serious dysfunctions with movement, speech, swallowing, and balance, as well as non-motor symptoms like depression, anxiety, sleep, and cognitive problems, significantly impacting daily activities and ultimately affecting the ability to live independently and maintain a good quality of life. As of now, PD remains pathologically and biologically unclear; and no disease-modifying treatment (DMT) current exist that can slow, halt, or reverse PD disease course. Prior neuropathological evidence indicates that PD pathology may begin up to two decades prior to clinical presentation–40–60% of dopaminergic neurons in the substantia nigra have been lost at time of motor symptom manifesting. Therefore, by the time PD is clinically diagnosed, neurodegeneration may be too advanced to be modified by therapeutic intervention. This has been recognized as a potential reason for the failure of many DMT trials. The pre-diagnostic or prodromal phases of PD (pPD) might represent a critical therapeutic window, during which molecular pathology may still be reversible or more responsive to intervention. In this context, this project aims at targeting the pPD phase to advance development of DMTs for early PD prevention. To this end, Aim 1 will develop an advanced AI-driven framework for harmonizing multimodal health data across PD research datasets, general-purpose biomedical databases, and large-scale EHR, and for building multimodal AI model for early prediction of PD. Aim 2 will develop causal learning-based trial emulation framework to identify and evaluate DMT repurposing candidates in PD, targeting the population with pPD. Aim 3 will leverage PD-derived iPSCs (in vitro) and mouse models (in vivo) for screening and validating the drug repurposing candidates.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Islet transplantation is a promising treatment for type 1 diabetes but suffers from significant islet loss with the standard intraportal liver infusion, consequently requiring large numbers of costly islets, often multiple transplants, and suboptimal achievement of long-term insulin independence. This is because islet isolation protocols result in regression of islet-specific endothelial cells (ISECs), with the loss of the critical, supportive islet vascular niche resulting in islet death. Furthermore, strategies to achieve islet engraftment in an extrahepatic site have been stymied by poor vascularization and lack of knowledge of how β-cell interaction with ISECs contributes to β-cell homeostasis. The overall goal of this project is to engineer ISECs to uncover the cellular cross-talk between -cells and their specialized vascular niche, to ultimately augment transplanted islet engraftment in the subcutaneous space. Each organ is vascularized by unique, specialized endothelial cells (ECs) that provide a tissue-specific vascular niche that supplies angiocrine factors key in choreographing organ homeostasis and repair. Indeed, there is growing evidence that a functional and physical interplay exists between specialized ISECs and -cells. Employing single cell analyses, we obtained a molecular signature of ISECs, identifying the novel transcription factor NKX2-3. This proposal will explore if vascularization of islets with NKX2-3+ ECs will facilitate engraftment, function, and survival of subcutaneously transplanted islets. ISECs have the additional critical feature of modulating the expression and migration of immune mediators, with macrophages being the most abundant pancreatic immune cell. In turn, these macrophages supply growth and immunomodulatory factors to sustain the integrity of -cells. Therefore, we hypothesize that induction of NKX2-3 in ECs confers these cells with the specialized properties of ISECs which, by supplying defined angiocrine factors and proper polarization of pro-reparative macrophages, support islet function. Accordingly, we will test the hypothesis that NKX2-3 is necessary for the specification of ISECs, which have the capacity to support islets in vitro and in vivo, through the following aims: Aim 1: Assess the impact on islet function of selective and conditional loss of NKX2-3 in pancreatic ECs. Aim 2: Examine the efficacy of enforced NKX2-3 expression in human umbilical vein endothelial cells in augmenting durable subcutaneous engraftment of transplanted islets. Aim 3: Decipher the mechanism by which NKX2-3, through macrophage polarization, coordinates pro-regenerative and anti-fibrotic islet innate inflammatory responses within the pancreas. The proposed training will guide and enhance my development in core competencies, including immunology, bioinformatics, and stem cell-derived islets, that will enable me to transition to research independence as a surgeon-scientist dedicated to improving islet transplantation outcomes. Weill Cornell Medicine is an ideal environment to execute this training plan due to its outstanding physical resources and its robust intellectual community of researchers with strong records of mentorship of early-stage investigators.

NIH Research Projects · FY 2026 · 2026-04

Acute radiation syndrome (ARS) is a multi-organ failure (MOF) syndrome, resulting from accidental exposure to irradiation and/or nuclear terrorism. Acute and delayed effects of acute radiation exposure (DEARE) can also be seen in the clinic after radiotherapy. While there are FDA-approved drugs for mitigating hematopoietic ARS (H-ARS), there are no treatment for repairing irradiated intestines, primary cause of death and a critical component of gastrointestinal ARS (GI-ARS). A common feature of radiation injury is damage to the microvascular capillary endothelial cells (ECs) that are not only organotypically programmed to modulate metabolism but also provide tissue-specific angiocrine growth factors that orchestrate intestinal repair and regeneration post-radiation. Indeed, intestinal-specific microvascular ECs (InSECs), defined by the expression of transcription factor NKX2-3, establish instructive vascular niches that are essential for maintenance, repair and regeneration of intestinal stem cells (ISC) of irradiated intestines. We show that InSECs choreograph the polarization of the monocytes into reparative IL1beta macrophages that could play a key role in resolving radiation injury. The objective of this proposal is to leverage reproducible in vitro extra-corporeal perfusable vascularized intestinal organoids models to test approaches to restore normal homeostatic functions of InSEC. Thus, we hypothesize that radiation impairs NKX2-3+ InSEC-derived instructive signals and retards recruitment and programming of intestinotropic macrophages that convey reparative signals for intestinal repair. Restoration of NKX2-3 transcriptional signatures in InSECs would reconstitute angiocrine and intestinotropic macrophage functions, essential for resolution and recovery after radiation injury. Delivery of pro-reparative factors such as amphiregulins and epiregulins supplied by macrophages and InSECs could mitigate radiation-inflicted intestinal injury. These hypotheses will be tested through executing the following specific aims: Aim 1: Develop and characterize GI-ARS injury models in human Gut-On-VascularNet organoid extracorporeal platform. Aim 2: To characterize the “instructive” angiocrine signaling pathways of InSEC on pro-regenerative monocytic polarization that mitigate radiation injury to HIOs. Aim 3: To screen for intestinotrophic radiation MCMs to mitigate GI-ARS by using the Gut-On-VascularNet platform. To this end, Rafii and Guha groups plan to capitalize on an engineered scalable near-physiological and reproducible extra- corporeal human Gut-on-VascularNet platform to study radiation injury in GI-ARS. Leveraging this platform, also allows for examining the role of radiation in perfusable microfluidic devices harboring NKX2-3+ InSECs vascularized intestinal organoids to screen for angiocrine factors and reparative macrophages to mitigate GI- ARS. The human Gut-On-VascularNet platform would bridge the gap between animal models and humans for the approval of radiation countermeasures for mitigation of GI-ARS under the FDA “animal rule” guidance.

NIH Research Projects · FY 2026 · 2026-04

Project Summary Glioblastoma (GBM) is an aggressive and incurable brain tumor marked by its high invasiveness, resistance to therapy, and universal recurrence. A key clinical challenge in GBM treatment is the diffuse infiltration of glioblastoma cells (GBCs) into the neighboring brain parenchyma, limiting the efficacy of current treatment plans. GBCs exhibit extensive genetic heterogeneity, with relapse often driven by subclones originally present but genetically distinct from dominant clones during diagnosis. Recent animal models suggest the heterogenous nature of GBCs’s ability to form interconnected networks, with a subset of GBCs becoming unconnected with a heightened invasiveness. Although multi-region whole-genome sequencing of patient samples suggests clonal invasion, technical limitations still obscure whether these migratory and invasive cells are truly (i) clonal in origin and (ii) driven by cell-intrinsic factors such as genetic alterations, or by cell-extrinsic influences from the tumor microenvironment (TME). Understanding how genetics, transcriptional states, and TME drive the evolutionary trajectory of invasive cells is key to disrupting their behavior. This investigation requires addressing two challenges: (1) linking large-scale migratory clones with genomic and transcriptomic data, and (2) uncovering how the tumor microenvironment (TME) influences subclonal evolution and invasive potential. To (1) track and characterize migratory clones, we will utilize the Landau lab’s expertise in joint single- cell whole-genome and transcriptome profiling across spatially distinct tumor regions. By constructing somatic phylogenies using single-nucleotide variants as natural lineage markers, we aim to trace the emergence and spread of invasive clones, providing a "temporal microscope" of tumor evolution. By integrating single-cell multi- region lineage trees with genomics and transcriptomics, we will gain an unprecedented lens of the evolution of invasive clones through time and identify the genetic alterations and molecular drivers of their aggressiveness towards recurrence. To (2) explore how the TME contributes to invasion, we will incorporate Slide-tags— spatially barcoded oligos—to map individual nuclei within their native microenvironment. This integration of high- resolution spatial with transcriptomics and cell lineages will allow us to assess how histological niches such as necrosis impose selective pressures on GBC subclones. Together, with three-dimensional layers of information for each cell, we can gain valuable insights into how the TME shapes the clonal architecture and cellular programming of GBCs to promote invasive phenotypes. This project will be ideal for me as a training scientist, given its use of novel single-cell technologies and my interest in somatic evolution, along with its direct clinical impact. With the mentorship of my sponsor, thesis committee, PhD program, and the support of this fellowship, I am confident I will be well prepared to pursue and achieve my goal of becoming an independent researcher capable of leading my own research group.

NIH Research Projects · FY 2026 · 2026-04

Project Abstract: Significant advances in antiretroviral therapy (ART) has enabled functional suppression of HIV, allowing individual individuals on ART to live relatively normal lives. However, viral rebound from a persistent reservoir of provirus-harboring cells following ART cessation demands a lifelong reliance on ART. ART-free control of this persistent reservoir therefore represents the current focus of HIV cure strategies. Recent work from our group and others has identified HIV-infected CD4 T cells that exhibit cell-intrinsic resistance to CD8 cytotoxicity, despite antigen expression and recognition. While identification of resistance factors previously annotated in cancer (such as overexpression of the granzyme inhibitor SERPINB9 and the anti-apoptotic protein BCL-2) has provided some insight concerning underlying mechanisms, the factors leading to this resistance phenotype remain largely unknown. To address this, we have developed a novel genome-wide screening platform that allows identification of genes involved in primary human CD4 T cell resistance and susceptibility to CD8 T cell cytotoxicity. In this proposal, we aim to maximize the potential of this newly developed CD4 resistance screening model (CRSM). Additional screens in different contexts will be conducted to further enrich the dataset. We will also employ a methodical pipeline for hit validation and therapeutic characterization. This pipeline will begin with promising hits identified in the initial screen, including the lipid metabolism-associated PPAR family proteins. This proposal will produce a robust functional genomic dataset thoroughly characterizing the mechanisms underlying CD4 resistance to CD8 killing. With these datasets, we envision the nomination of a range of promising targets for small-molecule sensitization of HIV-infected CD4s to CD8 killing.

NIH Research Projects · FY 2026 · 2026-04

Abstract. Variants of APOE are the major genetic risk factors for Alzheimer’s disease (AD). APOE has 3 common variants; APOE3 average risk, APOE4 high risk and APOE2 protective. This data, and in E2E4 heterozygotes, E2 cancels out deleterious effects of E4, led to the concept that gene therapy of a protec- tive APOE in the CNS would reduce the risk for AD in APOE4 homozygotes. Should the CNS gene ther- apy to treat APOE4 homozygotes be with APOE2 (the naturally occurring “protective” allele), or can we modify the CNS of APOE4 homozygotes with a more potent protective APOE? We focused on APOE Christchurch (Ch, R136S), a gain-of-function allele that protects against development of Tau pa- thology and clinical AD in the PSEN1/E280 Colombian family with early onset dominant AD. Using the AAVrh.10 neurotropic adeno-associated virus (AAV) capsid, we tested the hypothesis that AAVrh.10- mediated CNS delivery of the APOE2 allele with the Ch mutation (“E2Ch”), will provide superior protection against APOE4-associated AD amyloid and tau pathology compared to unmodified APOE2 (“E2”). The vectors were assessed in 2 mouse strains with humanized APOE4: APP.PSEN1/TRE4 “amyloid mice” and P301S/TRE4, “tau mice”. The novel E2Ch variant was more effec- tive than E2 alone to treat both the amyloid and tau pathology of murine AD in APOE4 background. Based on this and extensive safety data, we are ready to initiate a Phase I clinical trial of administration of AAVrh.10hAPOE2Ch to the CNS of APOE4 homozygotes with early onset AD. Aim 1 (UG3). Carry out a Phase 1A safety/dose-ranging clinical trial of CSF administration of AAVrh.10hAPOE2Ch to APOE4 homozygotes with early AD to determine the highest tolerable dose. Gain regulatory approval [FDA (IND), IRB, Biosafety, DSMB] to initiate the clinical trial. Manufacture clinical grade AAVrh.10hAPOE2Ch for Phase 1A. Vector administration to CSF via C1-C2, doses 5.7x1012 to 5.7x1014 total genome copies in 5, ½ log increments, n=3 participants each dose. Primary endpoint – safety parameters to determine the highest tolerable dose. Secondary endpoint – preliminary assessment of efficacy biomarkers and CSF levels of APOE2Ch. Aim 2 (UH3). Carry out a Phase 1B study at the highest tolerable dose from Phase 1A focused on exploratory assessment of efficacy parameters. Manufacture clinical grade AAVrh.10hAPOE2Ch for Phase 1B, CSF administration of the highest tolerable dose to n=10 early AD APOE4 homozygotes. Primary endpoint. Pre- and post-therapy evaluation of: target engagement (quan- tification of binding of CSF vector derived APOE to heparin); amyloid and tau PET scans; CSF levels of APOE4 and APOE2Ch, amyloid (Aβ42, Aβ40), tau (total tau, p-tau) GFAP, NfL, biomarkers; MRI CNS vol- umes and cognitive testing. Secondary endpoint – safety parameters.

NIH Research Projects · FY 2026 · 2026-04

Abstract. Human immunodeficiency virus type 1 (HIV-1) facilitates viral egress and ingress via the activity of the polyprotein, Gag. Gag is the sole facilitator of the identification, organization, and sequestration of the viral genome from the host cytoplasm to the plasma membrane, nucleating the assembly of a nascent virion. To produce infectious progeny, this process must occur with remarkable efficiency and specificity, despite the tremendous diversity among viral genomes. After budding, Gag is cleaved into its constituent domains by the viral protease, facilitating the intra-virion assembly of the conical viral capsid (CA) core (Core) and the compaction of the viral RNA genome by the viral nucleocapsid (NC). Upon entry, the Core is released into the cytoplasm and must translocate to the nucleus, traverse the nuclear pore, facilitate reverse transcription, while also shielding the viral genome and its reverse transcription products from the host immune system. The precise determinants and genetic constraints of infectivity by HIV-1 Cores remain incompletely identified and even less is known about HIV-1’s lentiviral relatives. The main goal of this project is to apply deep mutational scanning methodologies to understand the basic biology of the HIV-1 Core and potential heterogeneity therein. In the laboratory setting, single nucleotide substitutions within protein coding and non-coding regions of the HIV-1 genome have been demonstrated to have dramatic effects on viral fitness. Despite this, HIV-1 produces enough infectious progeny to remain viable in most infected individuals. And despite remarkable improvements in our ability to detect mutations in trace amounts of viral genetic material, the precise consequences of mutations within the HIV-1 genome have yet to be comprehensively investigated at scale. Therefore, Specific Aim 1 will seek to Identify the genetic determinants of HIV-1 Core assembly and infectivity. Similarly, Specific Aim 2 will extend this analysis to interrogate divergent lentiviral Cores, to characterize the more general constraints of lentiviral Core form, function, and adaptation to new intracellular barriers to infection. The results of this work could have implications for our basic knowledge of the cell biology of HIV-1 infection, the clinical treatment of HIV-1 infection, the prediction of drug-resistance mutations by HIV-1, and the development of next-generation lentiviral vectors for various applications in the laboratory and clinic. With the mentorship of my sponsor and thesis committee, the leadership of the Tri-I MD-PhD program, and the support of this fellowship, I believe that conducting the work described here will prepare leave me well prepared to take the next steps in pursuing a career as a physician-scientist.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY/ABSTRACT Chronic Hepatitis B Virus (HBV) infection affects nearly 300 million individuals and is a leading cause of hepatocellular carcinoma worldwide. HBV persistence relies on the establishment of 1) the viral covalently closed circular DNA (cccDNA) minichromosome which serves as that viral transcriptional template and 2) the expression of the regulatory protein HBx which prevents cccDNA silencing. Due to limited in vitro and animal models that recapitulate transient events like cccDNA formation early during infection, the mechanisms through which HBx recognizes and regulates cccDNA remain poorly understood. The David lab strives to better understand the molecular mechanisms of chromatin regulation in disease. Towards this end, we have pioneered novel platforms to study HBV biology with the first methods to reconstitutes scarless cccDNA with infectious capacity and full- length HBx protein for biochemical and biophysical studies. We discovered that HBx directly binds to chromatin to induce decompaction in vitro and found that cccDNA histone occupancy is required for HBx expression early during infection. My proposal extends these findings by seeking to elucidate host epigenetic regulation of HBV transcription (Aim 1) and HBx-mediated cccDNA recognition (Aim 2). My working hypothesis is that HBx directly recognizes and decompacts the viral minichromosome to allow host epigenetic machinery to distinctly control viral transcription. We have shown that host histone incorporation onto cccDNA is important for HBV transcription. Importantly, we found that pharmacological disruption of viral chromatin assembly leads to reductions in HBV transcription, antigen secretion, and viral replication in hepatocyte models of infection. The proposed Aim 1 will identify host histone variants and chromatin remodelers that influence HBV transcriptional control. First, I will probe for the presence of active transcription- associated histone variants deposited onto cccDNA during infection using chromatin immunoprecipitation. I will then determine the effect on viral chromatin assembly and transcription of the loss of host epigenetic regulators implicated in cccDNA chromatin regulation using a limited shRNA screen and auxin-inducible degron tagging. In Aim 2, I will use biophysical approaches, such as biolayer interferometry, to define the determinants of HBx- nucleosome binding. I will then investigate changes in viral chromatin compaction due to this interaction through Mg2+ precipitation and MNase protection assays. If validated in vitro, I will expand to testing this phenomenon in cellulo using dCas9 to fluorescently label cccDNA and ATAC-see to measure and visualize host- and cccDNA- specific changes in chromatin accessibility in the absence and presence of HBx. Overall, Aim 2 will reveal new insights into HBx’s role in viral chromatin regulation. Successful completion of this project will reveal mechanisms for how HBx modulates HBV chromatin and uncover novel therapeutic targets for silencing or eradicating cccDNA, improving epigenetic strategies to develop a functional cure for chronic HBV infection.

NIH Research Projects · FY 2026 · 2026-03

More than 80,000 new cases of adolescent and young adult (AYA) cancers are diagnosed annually in the United States. Alarmingly, the incidence of AYA cancers is expected to continue to rise in the coming years leading to a growing population of AYA survivors. Due to their young age at diagnosis, AYA survivors experience an extended survivorship during which they often contend with a spectrum of late and long-term effects of treatment, many of which have not been well-characterized in AYAs treated in the contemporary treatment era where novel therapies, including newer targeted agents, are increasingly used. Further, individuals representing different ages, diagnoses, and socio-economic backgrounds are often not well represented in AYA survivorship research. To address this gap, and given rates of AYA cancers are higher in New York City (NYC) compared to overall rates in the United States, we are proposing to establish the AYA Cohort to Enhance Survivorship in NYC (ACES- NYC), a prospective cohort that will enroll and engage 2000 AYA cancer survivors treated at three NYC cancer centers (Weill Cornell Medicine, Columbia University Irving Medical Center, and Memorial Sloan Kettering Cancer Center). In this cohort, we propose to characterize adverse short and long-term effects in the contemporary treatment era across a wide spectrum of AYA cancers, focusing on three central issues that uniquely impact AYA survivors: 1) fertility outcomes/reproductive health; 2) endocrine dysfunction (hypo/hyperthyroidism, obesity, diabetes); and 3) sexual functioning. Leveraging a mobile health application (app) platform that allows for linkage with the medical record, we will collect comprehensive treatment and clinical data, patient-reported data via serial surveys, sensor-based data collection enabled by the app, in addition to biospecimen acquisition. We will aim to: 1) determine the short and long-term impact of treatment on fertility outcomes, sexual health, and endocrine function, 2) determine demographic and clinical factors associated with underutilization of follow-up care related to reproductive, psychosocial, and endocrine health to inform timing and targets for intervention, and 3) develop, employ, and evaluate novel methods of data collection and engagement, in partnership with community-based stakeholders and patient advocates, to optimize recruitment and retention and to facilitate sharing of research results and survivorship resources. Together with community, patient, and research partners, our team will build and leverage ACES-NYC to shed light on numerous gaps in our knowledge of AYA cancer survivors’ short- and long-term experiences and needs, with a focus on improving survivorship care delivery and outcomes in this population.

NIH Research Projects · FY 2026 · 2026-03

Project Summary Mutations that reduce or alter the activity of repressive chromatin modifiers are frequent causes of neurodevelopmental disorders. The inaccessibility of the developing and the dynamic nature of neurogenesis so far have prevented an adequate understanding of how molecular pathologies arise downstream of the loss of specific chromatin regulators. This knowledge gap represents a hurdle for developing therapeutic interventions for neurodevelopmental disease. This proposal aims to combine targeted protein depletion with highly efficient directed differentiation regimens for human pluripotent stem cells to dissect gene regulatory functions of the autism-associated chromatin repressor EHMT1 during human cortical neurogenesis. Based on the extensive characterization of a novel, multipurpose (degradation/immunoprecipitation/visualization) degron allele, we hypothesize that interactions with cell type-specific co-factors allow EHMT1 to control the expression of stage- specific target genes during neurogenesis, resulting in the accumulation of molecular alterations and cortical neuron (CN) dysfunction when EHMT1 is lost from early development onwards. To systematically test this hypothesis, we will first determine whether molecular alterations caused by EHMT1 deficiency from earlier stages of neurogenesis onwards accumulate in CNs and to what degree dysregulated gene loci and CN function remain responsive to restoring physiological EHMT1 levels (Aim 1). We will then combine genomics, proteomics, and genetic approaches to identify how EHMT1, together with candidate recruiters and co-factors, regulates distinct gene loci at specific stages of cortical neurogenesis (Aim 2). Finally, we will expand our degron approach to dissect the functional interplay of different autism-associated H3K9 methylation regulators during human neurogenesis to identify interactions between these proteins that could be clinically exploited (Aim 3). Our experiments will determine currently unknown gene regulatory functions of disease-associated chromatin repressors at critical stages of human cortical neurogenesis. By generating mechanistic insight into how deficiencies of EHMT1 and other H3K9 methylation regulators introduce molecular pathologies in CNs, we anticipate revealing new opportunities for therapeutic interventions with specific neurodevelopmental diseases.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY Despite the broad use of antibody tools, their reliability and scalability remain major challenges for reproducible cellular and anatomical profiling in large tissue volumes, such as whole-mouse and human brains. In this project, we propose to develop an AI-assisted platform for recombinant antibody resources that meets the high standards required to advance brain research. By integrating our teams' complementary expertise in AI algorithms, structural analysis, antibody validation, and brain imaging, we aim to create a synergistic technical and resource platform for curating recombinant antibody tools, establishing a new paradigm for antibody applications in brain research. As part of the BICCN/BICAN effort, we have validated an extensive collection of commercial and open- source monoclonal antibodies using a high-content screening pipeline, providing a strong experimental foundation for recombinant antibody curation. However, the current experimental approach has limitations in efficiently selecting and prioritizing thousands of monoclonal antibody sequences for cost-effective conversion and validation. It also relies solely on existing sequences without effective optimization or design using structural information for diverse applications. Our integrated approach, leveraging our expertise in AI-based protein structure prediction, modeling, and simulation, will analyze antibody sequences to provide a comprehensive understanding of recombinant antibody structural properties, including antigen binding sites and affinities across target species. This will significantly accelerate the conversion and validation of recombinant antibodies. Additionally, we will continue refining our approach to enhance flexibility in optimizing and designing recombinant antibodies, tailoring them to specific epitopes and cross-species targets for diverse applications. We will also develop a curated database of recombinant antibody resource for easy reference and adaptation within the field. This database will include a comprehensive collection of validated recombinant antibodies, featuring their defined sequences, predicted antibody-antigen binding structures, 2D IHC data in mouse and human, and 3D whole mouse brain labeling datasets for key BICCN targets. To ensure broad accessibility, the database will be available through a user-friendly online portal. This integrated recombinant antibody platform, rAb-GenAI, will be open to incorporating advanced AI algorithms and continuously refined through iterative experimental feedback. Both our AI pipeline and recombinant antibody resource will be highly scalable and adaptable, maximizing cost efficiency to support consistent large-scale profiling, including human brain cohort mapping. Through this project, we aim to establish a new paradigm for antibody-based research, laying the foundation for brain-wide proteomic investigations across species and enabling studies on whole brain cellular and structural dynamics in health and disease.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY/ABSTRACT Access to high-quality primary care providers (PCPs), including physicians and advanced practice providers, is associated with better patient outcomes and is necessary to coordinate the complex needs of an aging population. Currently, primary care is experiencing significant corporate investment, which may have important implications for access to and quality of care in older adults. One key subset of investments has been from major pharmacy chains and retailers that own pharmacies. In the last five years, Walgreens invested over $6B to acquire majority stake in VillageMD, CVS acquired Oak Street Health for $10.6B, Walmart opened 51 Health Centers, and Amazon, which operates a mail-order pharmacy, acquired One Medical for $3.9B. In testing care models, these firms have acquired, opened, and closed many clinics, which are collectively referred to herein as retailer-owned primary care clinics. An important portion of these clinics focus on older adults, as firms see an opportunity to profit in value-based contracts in Medicare. Yet, the implications of retailer-owned primary care clinics for access to and quality of care in older adults is uncertain. The overall objectives for this proposal are to examine the implications of retailer-owned primary care clinics for access to and quality of care for older adults (age 65+). Through the proposed training plan, which includes formal coursework, workshops, and clinical shadowing, as well as a multi-disciplinary mentorship team of NIH-funded investigators, Dr. Kakani will gain clinical knowledge on primary care in older adults and on medication management of chronic disease, expertise in qualitative and mixed methods, and expertise in advanced statistical methods for causal inference. Dr. Kakani will use the knowledge gained through training activities to achieve the following specific aims: (1) Compare the area, PCP, and older adult patient characteristics of retailer-owned primary care clinics to other primary care practices to identify implications for access to and quality of care, (2) Evaluate the impact of retailer-owned primary care clinics on access to and quality of care for older adults, and (3) Characterize the perspectives of health care executives and PCPs of retailer-owned primary care clinics and specialist physicians treating patients using retailer-owned primary care clinics regarding factors impacting care delivery for older adults. This application is innovative because it will be the first study of retailer-owned primary care clinics and will leverage a mixed methods approach to provide nuanced insights into the implications of retailer-owned primary care clinics for older adults. The proposed research is significant because it will provide new, critical evidence on the impact of retailer-owned primary care clinics, which will inform innovators and policymakers seeking to reform and improve access to and quality of primary care in an aging population, and regulators of mergers and acquisitions in health care. This to access K01 Award will also provide Dr. Kakani with the training and mentoring needed become n R01-funded independent investigator leading a research program on primary care and medication and management in older adults. a

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY The proposed project encompasses a comprehensive research plan to support the development of my career as an independent scientist focused on investigating the assembly and conformational dynamics of neuronal integral membrane receptors, and how structure is related to receptor activation and regulation. This biophysical mechanism-focused approach to receptor function will guide promising therapeutic development. Background: The neurotrophic receptor tropomyosin kinase receptor B (TrkB) is a member of the receptor tyrosine kinase (RTK) superfamily of integral membrane proteins which receive extracellular signals and initiate intracellular signaling cascades across the cellular membrane. Canonical RTK activation is characterized by a transition from a monomeric receptor to a dimeric receptor following ligand binding. However, receptor assembly studies on EGFR, an extensively studied RTK, demonstrate that this activation pathway is much more complex than initially thought as an ensemble of ligand-dependent conformations have been observed. Despite the role that TrkB plays in neurodegenerative, psychiatric, and oncological disease, no effective therapeutic agent that targets TrkB is available due in part to the overwhelming lack of structural information relating to receptor function. Specifically, the field currently lacks an understanding of TrkB dimeric assembly and how this relates to intersubunit and intrasubunit conformational dynamics across domains. Specific Aims and Research Design: The proposed study investigates TrkB assembly, structure, and conformational dyanmics using 1) single-molecule and ensemble fluorescence-based binding methods to investigate the molecular determinants of TrkB dimerization 2) cryogenic electron microscopy (cryoEM) to assess the structural basis of TrkB dimeric assembly and allosteric activation. My in vitro and structural findings will be validated and extended by cellular and functional assays. Together this work will guide the development of a K99/R00 proposal aimed to apply the approaches and findings from this F32 research program broadly to other neuronal integral membrane receptors and to pursue the functional consequences of TrkB receptor assembly and conformational shifts in in vivo models. Training and Mentoring: My training goals are supported by 1) my sponsor Dr. Levitz and my assembled team of carefully selected consultants and collaborators with a range of expertise in membrane protein biophysical and biochemical research, 2) a flourishing institutional environment at Weill Cornell Medical College, and 3) scientific meetings, seminars, and planned publications. Impact: The invaluable experience gained during this award will serve as the foundation for my independent career as a protein biophysics researcher and for the development of a novel program of membrane protein research.