New York University School Of Medicine

NIH Research Projects · FY 2025 · 2025-08

Adaptive randomized clinical trials are critical in infectious disease research, offering flexibility to adjust sample sizes, introduce new interventions, discontinue ineffective treatments, and target specific subgroups to enhance treatment efficacy. This adaptability is particularly valuable in rapidly evolving public health crises, such as the development of treatments for emerging infectious diseases like COVID-19. However, adaptive trials present significant challenges, including unclear inferential targets, statistical biases from temporal and spatial variability, complexities in handling dynamic data structures, and an increased risk of false-positive findings. These concerns are reflected in recent FDA guidance on estimands, which emphasizes the need for clearly defined inferential targets, and on adaptive designs, which acknowledges that statistical bias in adaptive trials remains an understudied issue. Despite these recognized challenges, current research lacks a principled framework for structurally representing and unbiasedly estimating causal effects in adaptive trials. This project will develop a structural causal framework for adaptive trials, leveraging modern causal inference and statistical techniques alongside secondary data from the Adaptive COVID-19 Treatment Trial (ACTT)—an adaptive trial evaluating novel therapeutics in hospitalized COVID-19 patients—to enable transparent, efficient, and statistically unbiased estimation of causal effects. To achieve this, we propose the following specific aims: Aim 1: Develop a structural causal approach that deals with temporal variability. Aim 2: Extend our framework to handle spatial variability. Aim 3: Expand our framework to handle complex data structures, including failure-time and missing data, while dealing with false-positive results. Our project aligns with NIAID’s mission by advancing key methodologies for infectious disease clinical trials, particularly in adaptive designs for pandemic response, emerging pathogens, and the development of antiviral treatments. While our primary focus is on infectious disease trials, our methods have broader applicability to other disease areas, such as schizophrenia. We show this by also leveraging secondary data from schizophrenia studies, including the DECIFER trial, the RAISE study, and the EPINET study. RELEVANCE (See instructions): This research aims to improve how adaptive clinical trials are designed and analyzed. By developing methods that address key challenges in adaptive trials, our work will help ensure more accurate and reliable results, ultimately leading to better treatments and public health responses to emerging infectious diseases.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY / ABSTRACT: DECODING POSITIONAL EPIGENETIC MEMORY Hox gene expression is crucial for patterning the main body axis of all bilaterian species. In a classic epigenetic memory fashion, positional identity is acquired early in development and is inherited through cell lineage via Hox expression. Misexpression of even a single Hox gene can lead to homeotic transformations or diseases like cancer, making stable and accurate Hox positional epigenetic memory essential for proper cell differentiation and disease prevention. Building on a decade of seminal contributions to the field, our research aims to decipher how stable Hox positional identity is generated. In mammals, the Hox genes are organized into four chromosomal clusters, each containing a subset of 13 genes within approximately 100 kb. Polycomb Repressive Complexes (PRCs) decorate and suppress Hox clusters in undifferentiated cells. Transient patterning signals such as retinoic acid partition Hox clusters into active (PRC-removed) and repressed (PRC-high) domains at early differentiation stages, establishing precise and stable boundaries that convey positional identity across cell generations. We made three critical contributions to the accepted Hox positional epigenetic memory model. First, we identified that patterning signals result in the binding of activating transcription factors to Hox cluster domains. Second, we discovered that this binding results in PRC eviction from CTCF-established chromatin boundaries. Lastly, we demonstrated that the Hox cluster is sufficient to establish positional identity. In other words, a 100 kb Hox cluster engages with the three elements required for positional memory: repression (PRC recruitment), activation (signaling transcription factors), and boundary formation (CTCF). The ability of CTCF to engage in boundary formation is an active area of research and the best-understood component. However, there is a knowledge gap in the mechanisms that control activation and repression: Challenge 1: Activation. The current working model cannot explain how Retinoic Acid signaling evicts PRC while initiating a progressive colinear Hox gene transcription. Further, the model lacks definitive proof of the physical distribution of the activating Retinoic Acid Receptor and CDX2 factors along Hox cluster domains. We will challenge the Hox regulatory paradigm to the core by building novel Hox regulatory units with synthetic DNA technology. Challenge 2: Repressors. Although it is among the earliest examples of PRC repression, there is no mechanistic understanding of how Hox clusters recruit PRC in mammalian cells. With our powerful synthetic DNA approach, we will identify the mammalian Hox polycomb recruitment element (PRE). By merging synthetic DNA, new genomic engineering tools, and stem cell differentiation systems, we aim to understand positional memory through Hox chromatin boundary formation and build the smallest genetic unit sufficient to establish and sustain positional epigenetic memory.

NIH Research Projects · FY 2025 · 2025-08

Summary Neocortical layer 1 (L1), the “crowning mystery” of David Hubel, is the most superficial layer of the cortex. It is a major target of high order cortico-cortical and thalamocortical projections that carry “top-down” or “feedback” information, such as behavioral saliency, expectations, predictions, and memories. This top-down information is integrated with bottom-up sensory input received by the pyramidal cells (PCs), the output neurons of the cortex, to generate percepts and drive context-dependent behavior. A detailed understanding of how top-down and bottom-up signals are integrated is necessary to understand functions such as sensory perception, learning and decision making. This is also of significant medical relevance, as disruptions of this integration are thought to underlie various neurocognitive disorders such as autism and schizophrenia. L1 is unique among neocortical layers in that it lacks excitatory neurons but instead contains the distal (tuft) dendrites of the pyramidal cells (PCs) located in deeper layers. These dendritic compartments receive the diverse top-down projections arriving in L1, allowing their integration with the feedforward sensory input arriving at the basal dendrites of the PCs. L1 also contains a complex specialized population of GABAergic interneurons (INs) the only neurons in this layer. L1 INs modulate how top-down information is relayed to the tuft dendrites of the PCs. However, the precise circuit mechanisms through which these INs regulate the processing of top-down signals is poorly understood. Unlike INs in other layers, which have been the focus of extensive research, L1 INs are distinct, and their functional roles are less well characterized. This knowledge gap has persisted largely because of the lack of molecular genetic tools that have facilitated the study of neuronal circuits in other layers. Recent efforts by us and others have provided genetic reagents that facilitate the identification and manipulation of three of the four IN populations of L1: the NDNF neurogliaform cells (NGFCs) and canopy cells, as well as to VIP INs, an interneuron population that has low abundance in L1, but is highly enriched in L2/3 and contains prominent dendrites in L1. However, we still lack access to the fourth IN population, called a7 INs due to their prominent expression of a7 nicotinic receptors (a7 nAChRs). a7 INs are also unique to L1, are key targets of long-range projections of L1 and we hypothesize they define a novel L1 disinhibitory system. To advance our understanding of signal integration in the neocortex we are studying signal processing in L1 in an associative learning task, for which we plan to submit a BRAIN Circuits Projects R01. To prepare for this submission, in this R34 Targeted BRAIN Circuits Planning Project we plan to develop tools to gain genetic access to the L1 a7 INs. In Aim 1, we propose several strategies to develop mice to target the a7 IN population (Aim 1A) and utilize these mice to investigate the efferent connectivity of a7 INs unto L2/3 and L5 PCs and INs (Aim 1B). In Aim 2, we generate a conditional knockout of the a7 nAChR in L1 a7 INs to obtain support for the hypothesis that the fast cholinergic modulation of these neurons by these receptors is essential for associative learning in our behavior.

NIH Research Projects · FY 2025 · 2025-08

Project Summary: Ubiquitylation, the speci3ic addition of ubiquitin groups to targeted proteins, dictates a multitude of cellular pathways crucial for maintaining cell homeostasis, including cell cycle pathways. Cyclin D, a key regulator of the cell cycle, predominantly modulates the progression from the G1 to S phase through the activation of its partners cyclin-dependent kinase 4 and 6 (Cdk4/6). Its expression is tightly controlled to prevent aberrant cellular proliferation. The ERAD (Endoplasmic reticulum associated degradation) system plays a fundamental role in protein homeostasis, speci3ically by the identi3ication and targeting of misfolded or aberrant proteins located within the ER. ERAD substrates are selected by molecular chaperones for degradation by the ubiquitin–proteasome machinery. Disruptions in the ERAD operation can activate cellular stress responses, possibly leading to disorders like neurodegenerative diseases and cancers. Intriguingly, in differentiated cells such as neurons, aberrant activation of the cyclin D-Cdk4 complex has been linked to protein aggregation and degenerative conditions. This multidisciplinary research proposal aims to broaden our understanding of cell-cycle regulated processes affecting protein homeostasis. We focus on a novel cyclin D dependent regulation implicated in orchestrating the ERAD. Our overarching aspiration is to bridge the knowledge gap between protein aggregation and cell cycle anomalies. To this end, we propose the following holistic studies: Aim 1: We plan to employ advanced structural techniques to understand how a D-type cyclin-dependent phosphorylation modulates the interface between key ERAD effectors. Additionally, in vitro studies will be conducted to understand how phosphorylation impacts the proteins’ structural and functional attributes. Aim 2: The project seeks to elucidate the mechanism of action of the novel cyclin D-dependent regulation in the ERAD degradation machinery. We aim at investigating its role in the breakdown of speci3ic substrates identi3ied through unbiased screening methods. To facilitate this, we have engineered cell lines that enable controlled expression of 3luorescently labeled reporter substrates in the presence of inducible cell cycle regulators. Aim 3: Our 3inal objective involves investigating the role of the cell cycle regulation in the protein degradation machinery in shaping the proteomic landscape at membranal organelles. A combination of methodologies such as live-cell imaging, proximity ligation techniques, and compartment-speci3ic sub-fractionation will be employed. These studies will be validated using a genetically engineered mouse model. By achieving these objectives, we aim at 3illing existing gaps in our knowledge about the intricacies of cell cycle regulation and protein degradation. The outcomes of this research are expected to offer invaluable insights into diseases characterized by protein aggregation.

NIH Research Projects · FY 2025 · 2025-08

Project Summary NYU Grossman School of Medicine and Erasmus University Medical Center jointly propose to evaluate how organophosphate pesticides, glyphosate, phthalates and bisphenols contribute to renal risks in childhood and adolescence. The incidence of chronic kidney disease (CKD) is steadily rising and synthetic chemicals are increasingly understood to contribute to acute and chronic kidney injury. Case-control studies of populations with high incidence rates have identified pesticide and herbicide exposures as risks, raising the question whether developmental exposures may be even more impactful. Our own studies of children with CKD (R01DK100307) have revealed modest declines in kidney function with increasing phthalate and bisphenol exposures, accompanied by increases in oxidative stress. However, these findings do not contribute to our understanding of the origin of CKD. A major limitation is the failure of our and other studies to account for the developmental biology of the kidney and strong influence of perinatal/infant factors. The premise of the present proposal is that intrauterine inflammatory processes disrupt nephrogenesis and that environmental chemicals also impair renal parenchymal growth longitudinally during gestation and postnatal development via oxidant stress. We further interrogate this hypothesis by examining phthalates, bisphenols, glyphosate and organophosphate (OP) pesticides as modifiable risks. We will test these hypotheses in Generation R First, a prospective, longitudinal multi-ethnic birth cohort with existing measurements of phthalate and bisphenol exposure at three time points in pregnancy and three time points in childhood (5-6, 9-10 and 13-14 years) in 1405 mother-infant pairs (funded by R01ES022972 and R01ES032826). Organophosphate (OP) exposures were measured in the same time points in pregnancy in a subsample of 776 (ZIAES103314). We therefore propose to: add measurements of glyphosate and oxidative stress exposure in pregnancy and childhood at the same six time points (n=1312 children with at least one measure of kidney size, eGFR or albuminuria); complete the measurements of OP in pregnancy and childhood to permit mixture analyses that also consider phthalates, bisphenols and glyphosate; and evaluate glomerular (albuminuria) and tubular injury (KIM-1, NGAL) as well as functional renal mass (urinary epidermal growth factor, EGF) at the four time points in childhood. Leveraging Generation R First for the proposed work presents substantial cost-efficiency, with available measurements of kidney size, eGFR and albuminuria at ages 5-6, 9-10 and 13-14, and planned measurements at 17-18 years of age. The proposal couples the expertise of an internationally renowned environmental pediatrician (Trasande) with a life course epidemiologist (Jaddoe) who is PI of Generation R.

NIH Research Projects · FY 2025 · 2025-08

The initial interaction between a virus and a human cell represents the critical first step in viral infection, making its prevention a primary focus in antiviral therapy. The viral entry process remains largely unexploredfor orthopox viruses, which caused an escalating number of outbreaks in recent years. In 2022, the mpox virus (MPXV) clade 2b instigated a multi-country outbreak involving both endemic and non-endemic countries, leading to an international health crisis (WHO) and a public health emergency declaration (U.S.). In 2023, the Democratic Republic of Congo (DRC) experienced an outbreak of the more lethal MPXV clade 1 ( with a fatality rate of up to 10%) linked to sexual transmission. Individual proteins involved in the multi-protein entry/fusion process and targets for neutralizing antibodies were described, yet a comprehensive understanding of orthopox virus entry is missing. This leaves a blind spot in understanding the sequential procedure of viral entry, the intricate interactions among the numerous proteins involved, and the implications for vaccine design and therapies. Here we propose to elucidate the molecular process of orthopox virus entry through side-by-side comparison of MPXV and vaccinia virus (VACV) using an integrative approach that combines methods from virology, proteomics, and immunology. Aim 1: Identifyand characterize the interactions of virus andcellproteins during attachment and entry, and determine antibody accessibility to viral entry proteins. We will conduct in vitro orthopox virus infections of Vero E6 cells to understand interaction processes during viral entry. We will analyze multiple time points within the initial 30 minutes post-infection to track sequential interactions between viral and cellular proteins during the attachment and entry phases. Coupling trifunctional crosslinking of cellular and viral proteins with mass spectrometry will reveal virus–host interactions. We will use RNA interference to confirm the relevance of the identified cellular receptors for orthopox virus entry and infection. We will classify cellular proteins by enrichment analyses and pathway networks. Furthermore, we will use: 1) crosslinking to polyclonal antibodies (purified fromsera of mpox convalescent patients and vaccinees), and 2) quantitative analysis of the biotinylated viral surface proteins to identify antibody-accessible entry proteins on MPXV and VACV virions. Aim 2: Map antigenic properties of the viral proteins from the orthopox virus entry cascade. We will use serum antibodies from mpox convalescent and/or vaccinated individuals, collected during the 2022 NYC mpox outbreak, to study recently reported and newly identified viral proteins of the entry cascade. Together, these approaches will address fundamental questions in orthopox virology and decipher the viral entry process and antibody exposure during entry at the molecular level. The insights gained will provide unprecedented detail to help understand how orthopox viruses infiltrate cells, providing critical knowledge to design next-generation vaccines and treatments.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Lung cancer is the leading cause of cancer related deaths worldwide. For advanced lung adenocarcinoma, the major subtype of lung cancer, the standard of care treatment is immune checkpoint blockade which enhances anti-tumor immune responses. Unfortunately, many patients do not respond to the mechanisms for unclear reasons. LKB1 mutations are found in approximately 20% of lung adenocarcinoma cases. Patients with these specific tumor mutations have poor responses to immune checkpoint blockade and overall worse survival. Currently there are no targeted therapies specifically for LKB1 mutant lung adenocarcinoma. Here I propose to use genetically engineered mouse models, which faithfully recapitulate human lung adenocarcinoma, to define how these specific tumors interact with the microenvironment. We have previously shown that LKB1 mutant tumors, through an autocrine LIF signing pathway, promote tumor dedifferentiation and, through expression of an inflammatory signature, recruits immunosuppressive myeloid cells. However, we have not delineated the signaling mechanism in which LIF promotes dedifferentiation. Furthermore, while the role of immunosuppressive myeloid cells impairing CD8 T cells is well characterized, little is known about these myeloid cells signal to the tumor cells themselves. I hypothesize that there are two cytokine signaling loops in LKB1 mutant tumors: one in which autocrine LIF signaling promotes tumor dedifferentiation and an increase in inflammatory markers promoting neutrophil and macrophage recruitment and a second cytokine loop where myeloid cells release Il-1β to amplify the inflammatory signal of LKB1 mutant tumors. In Aim 1 I will use Lkb1 mutant lung cancer mouse models to interrogate the autocrine LIF signaling loop using scRNA-seq and scATAC-seq to identify signaling mediators for tumor dedifferentiation and inflammation. I will then perform CRISPR/Cas9 editing to generate knockouts to validate those findings. In Aim 2 I will initiate Lkb1 mutant lung tumors with knockout of the receptor for IL-1β and measure changes to the immune microenvironment as well as tumor growth. I will then investigate the therapeutic role of IL-1β neutralization in the context of Lkb1 mutant tumors. The proposed study will be led by Dr. Ray Pillai, a Clinical Instructor at NYU Grossman School of Medicine under the co-mentorship of Drs. Thales Papagiannakopoulos and Sergei Koralov who have combined expertise in cancer mouse models, immunology, and metabolism. Both mentors have created a training plan where Dr. Pillai will transition to an independent investigator. An advisory committee, consisting of Drs. Kwok-Kin Wong, Dan Littman, and Mark Philips, will also help guide Dr. Pillai on his path to becoming a physician scientist.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY: In this grant proposal, we aim to develop the L7Ae protein as a therapeutic for treating Orthomyxoviridae family infections. Our approach includes three aims: (1) Investigating the mechanism of action of L7Ae on various family members, focusing on its interaction with the conserved k-turn recognition domain in the L30 RNA- binding proteins, to identify key viral targets and understand how L7Ae disrupts the viral life cycle. (2) Characterizing a recently developed L7Ae transgenic mouse model that shows normal development but resistance to influenza A virus, providing an independent platform to study L7Ae's effects on host and virus biology. (3) Assessing the administration of recombinant L7Ae in vivo by introducing it into mouse lungs and evaluating lung function parameters and histological analyses to examine its impact on lung morphology and inflammation, thereby evaluating potential adverse effects on the host. Our ultimate goal is to generate preclinical data on L7Ae-mediated treatment for Orthomyxoviridae infections, aiming to advance it towards a clinical usage.

NIH Research Projects · FY 2025 · 2025-07

SUMMARY Staphylococcus aureus infections, especially methicillin-resistant strains (MRSA), lead to high morbidity and mortality rates in hospitals. Currently, hospitals primarily rely on clinical cultures for MRSA surveillance, which often fails to accurately detect transmission events due to their inability to assess genetic relatedness among bacterial strains. Using whole-genome sequencing (WGS) for early outbreak recognition is crucial for enhancing infection control measures that will reduce the morbidity, mortality, and dissemination of pathogenic genetic variants. However, our preliminary hospital-wide WGS analysis indicates that detecting MRSA transmission in real-time remains challenging because individuals involved in asymptomatic transmission events are not sampled. Consequently, asymptomatic acquisitions often remain unnoticed until readmission or subsequent screening, delaying timely intervention and effective cluster management. Timely detection of transmission events is essential to stop the spread of MRSA and prevent outbreaks. To address the issue at the implementation level, we propose a two-pronged approach. In Aim 1, we will integrate weekly colonization sampling with hospital-wide genomic surveillance to analyze phylogenetic relationships among S. aureus strains. This will help establish a method for detecting real-time transmission events in high-risk hospital wards. Aim 2 focuses on developing predictive models that combine patient, bacterial, and contact network characteristics linked to in-hospital transmission events. These models will synthesize demographic, spatiotemporal, and genomic data to optimize sampling intervals and, ultimately guiding future studies of real- time targeted interventions. We will also compare the cost-effectiveness of different sampling strategies. By enhancing our understanding of transmission dynamics and personalized predictive modeling, this research will shift hospitals from reactive to proactive management of S. aureus transmission clusters. The results will also provide crucial inputs to inform the design of future randomized trials to evaluate these strategies’ effects on morbidity, mortality, and cost. These insights are expected to have broad implications for the surveillance and management of other hospital-acquired pathogens, ultimately contributing to better patient outcomes and more efficient use of healthcare resources.

NIH Research Projects · FY 2026 · 2025-07

Selective neuronal vulnerability is a key feature of Alzheimer’s disease (AD), which also presents with other pathological features such as accumulation of intracellular neurofibrillary tangles and extracellular amyloid- β deposits. It has been known for decades that the basal forebrain cholinergic neurons (BFCNs) are severely affected in AD and highly vulnerable to early tau accumulation. The molecular mechanisms underlying pathophysiologic changes in BFCNs remain largely understudied. An overarching goal of this proposal is to reveal the interplay between tau pathology and cholinergic dysfunction, two key pathological features during the progression of AD. We will test the hypothesis that pathological tau causes BFCN dysfunction, which leads to altered cholinergic modulation of cortical function. By leveraging human induced pluripotent stem cell (iPSC)- derived BFCNs and a novel mouse model with restricted tau expression in BFCNs, we will assess the molecular and functional impacts of pathological tau accumulation in BFCNs at the neuronal, network, and animal levels. Knowledge obtained through the proposed study should have broad implications for clarifying tau pathogenesis in cholinergic dysfunction and degeneration, and inform potential therapeutics to improve the resilience of BFCNs to treat AD and related tauopathies. Research Projects: In this proposal, we will first investigate the biochemical underpinnings and functional alterations of BFCNs with pathological tau accumulation using an iPSC-derived cell model in Aim 1. Subsequently, Aim 2 will systemically examine tau spreading from the nucleus basalis BFCNs to their projected cortical areas, which may be modulated by age, tau species and neuronal activity, using a mouse model with restricted BFCN tau expression. Lastly, Aim 3 will assess the cholinergic modulation of cortical function, as well as behavioral outcomes in the mouse model with tau accumulation in nucleus basalis BFCNs. These experiments will help to dissect the molecular pathways that are altered in nucleus basalis BFCNs with tau pathology, and elucidate the functional consequences of tau accumulation in these BFCNs. Candidate Development and Environment: The training plan outlines the approaches that the candidate will employ to engage the rich resources available in her laboratory and the neuroscience community at NYU Langone Medical Center. During the K award, the candidate will expand her expertise in cellular and molecular neuroscience, in vitro disease modeling, and functional studies. Additionally, she will gain expertise in proteomics, animal behavioral assessment, and in vivo brain circuitry studies. The candidate has assembled a team of mentors, advisors and collaborators for the proposed project. This will ensure successful completion of the studies by leveraging her unparalleled environment and resources. Overall, this project will ultimately enhance the candidate’s career goal of building an independent research program to study the pathogenesis of AD and develop effective therapeutics to treat this disease.

NIH Research Projects · FY 2025 · 2025-07

Homelessness and housing instability are strongly linked with overdose risk. Permanent supportive housing (PSH)—subsidized housing paired with support services—is a key part of the national strategy to end homelessness. PSH tenants face high overdose risk due to a confluence of factors. Despite this risk, little past research has examined overdose prevention in PSH or even as related to housing more generally. We propose a multi-sector, community-partnered, stepped wedge randomized controlled trial (RCT) to evaluate the impact of a technical assistance (TA) intervention designed to support PSH agencies in sustainably implementing evidence-based practices to reduce tenant overdose and improve tenant health more broadly. The intervention will be delivered to PSH agencies in New Jersey and New York by a collaborative multi-sector team led by community organizations with national footprints. The intervention addresses not only individual tenant behavioral risks but also acts at the larger agency and building levels. The study includes a UG3 planning phase and a UH3 trial phase, with milestones defined for each phase. In UG3 Aim 1, we will conduct focus groups with PSH leaders, staff, and tenants to refine an overdose prevention TA package (the intervention) that is feasible, acceptable, and appropriate for varied PSH types across New Jersey and New York. In UG3 Aim 2, we will conduct preparatory activities for the RCT, including site selection, data agreement execution, data collection instrument and analysis plan finalization, and IRB approval. In UH3 Aim 1, we will conduct a stepped wedge RCT to examine the impact of the TA intervention on PSH agency implementation of overdose prevention practices and downstream tenant overdose, substance use, and other health-related outcomes. Outcomes will be assessed using surveys, administrative Medicaid and PSH data, and PSH agency records. In UH3 Aim 2, we will identify implementation successes and challenges, and explore pathways between the intervention and tenant overdose risk and related health outcomes, using qualitative interviews with PSH tenants and staff. In UH3 Aim 3, we will integrate the results of UH3 Aims 1–2 using a convergent mixed methods design, and disseminate findings to ensure translation of the research into practice and policy locally and nationally. It is expected that findings from this research will be immediately actionable and will also be broadly applicable to other housing settings and homeless shelters. Ultimately, we aim to reduce overdose and improve related health outcomes by focusing on a critical yet understudied aspect of overdose prevention with national relevance. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions for the overdose epidemic, including opioid and stimulant use disorders. The NIH HEAL Initiative bolsters research across NIH to address the national opioid public health crisis and improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT Cannabis use in the U.S. has surged, driven by widespread legalization and accelerated by the COVID-19 pandemic. Over 55 million U.S. adults are current users, and approximately 30% will develop cannabis use disorder (CUD). Despite the urgent need for effective treatments, no FDA-approved options exist, and current interventions have proven insufficient, underscoring the necessity for innovative and accessible solutions. We propose a novel, home-based intervention combining transcranial direct current stimulation (tDCS) with mindfulness meditation, targeting the dorsolateral prefrontal cortex—a key region in the neural circuitry underlying the addiction cycle. Delivered via the ElectraRx telehealth portal, this approach is both cost-effective and widely accessible. Our extensive preliminary data, involving over 600 patients and 17,000 home-based tDCS sessions, along with our pilot study of this intervention in CUD, demonstrate its feasibility and potential. However, further development and testing are required to advance it to clinical use. The initial UG3 phase (Years 1-2) will involve developing a tDCS home-based administration system by Soterix Medical Inc. (SMI), tailored specifically for the intended population and environment ("ElectraRx-CARES"). We will then refine and validate the tDCS-mindfulness intervention through a double-blind, sham-controlled feasibility trial with n = 46 adults with CUD. If at least 50% of participants complete 70% of the sessions, we will advance to the UH3 phase, where a large-scale RCT enrolling n = 192 participants with CUD to assess the intervention's efficacy in reducing cannabis use and withdrawal symptoms. Follow-up assessments at three months will evaluate the persistence of these effects. FDA guidance on device development and data collection will be obtained throughout. Finally, we will optimize intervention delivery, iterate the device, and advance the regulatory process, including pursuing Breakthrough Device Designation. This scalable intervention addresses a critical public health need and represents a promising step forward in treating the rapidly growing problem of CUD.

NIH Research Projects · FY 2025 · 2025-07

Community-Engaged Approach to Supporting Patients with Type 2 Diabetes at Risk for Depression Background: Patients with Type 2 diabetes (T2D) are at increased risk for depression (60%), and patients with depression have a 32% increased risk for T2D. The resulting comorbidity has significant implications for disease burden, healthcare costs, and management of these disorders. In NYC, prevalence of diabetes ranges from 7.1%-14.4%, and T2D patients are twice as likely to have symptoms of depression. This comorbidity is more prevalent by sociodemographic characteristics such as sex, education level, income, and English proficiency. Given significant barriers to accessing medical and mental healthcare services (e.g., affordability, accessibility) there is an urgent need for interventions to comprehensively address this comorbidity. Building on an innovative and successful community health worker (CHW)-led T2D intervention at NYU Grossman School of Medicine (DREAM Initiative - R01DK11048), this K01 proposal will adapt this CHW-led T2D intervention to include mental health and digital health components, and test the feasibility and acceptability of this intervention to support participants with T2D who are at risk for depression. In this proposed pilot intervention, CHWs will provide tailored health education to participants on management of T2D and depressive symptoms using the DREAM intervention and adapted content from the WHO’s Group Problem Management+ psychological intervention program, and facilitate referrals to mental healthcare services. A CHW facilitated text message discussion group will reinforce health messages between health education sessions, and a wearable fitness tracker will be given to participants to monitor daily fitness and sleep patterns. Specific Aims: 1) Examine factors associated with distress, poor mental health, and comorbid T2D and depression among participants to inform pilot study structure and content; 2) Adapt CHW-led T2D intervention to include mental health and digital health components using a trauma informed care approach and the ecological validity model; and 3) Evaluate the feasibility and acceptability of a pilot CHW-led intervention to support participants with T2D who are at risk for depression. Training: The mentored training plan will include didactic and experiential learning opportunities. Primary training topics will include clinical management of diabetes, clinical T2D research, trauma informed care, mental health, digital health, and grant writing. The training will include formal coursework, workshops, seminars, directed readings with mentors, and participation in mentor study meetings.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract Despite great strides in achieving short-term weight loss success, long-term weight loss maintenance (WLM) remains a challenge. Time-restricted eating (TRE), a form of intermittent fasting, is an untested bio-behavioral strategy for WLM. The goal of this Pathway to Independence Award (K99/R00) is to accelerate the candidate's transition to an independent investigator with focused expertise in the development and evaluation of weight management strategies during WLM. In the K99 phase of this award, the candidate will obtain training in expertise in behavior maintenance theories and methods, mixed methods and qualitative analyses and appetite physiology, and conduct pilot studies to develop and test the intervention approach. The candidate will conduct a pilot randomized controlled trial (n=40 total; 20/arm) to test the feasibility, acceptability, and adherence of two TRE interventions (≤6 h and ≤10h eating windows). Participants will include adults (25–65y, BMI>20.5 kg/m2) recruited from 2 nonsurgical weight loss programs who successfully attained a weight loss of ≥5% initial body weight. Prior to randomization, participants will undergo a run-in phase that includes a 4-week weight stabilization period. The candidate will evaluate the feasibility and acceptability in terms of recruitment, retention, and adherence to the eating window using a mixed-methods design (Aim 1). The candidate will perform qualitative interviews (n=12/arm) following completion of the TRE to capture participants' experience with TRE and determine individual-, family-, and neighborhood/community-level barriers and facilitators to each TRE intervention, such as motivation, cultural factors, occupation, and family structure (Sub-aim 1) that will be used to strengthen the intervention. In the R00 phase, the candidate will use acquired skills and training to test the optimum TRE intervention identified from the K99 phase. In a randomized controlled trial, the candidate will assess the efficacy of the selected TRE on body weight regain (Aim 2a), and fat mass regain (Aim 2b) at 12 months. The candidate will recruit adults with recent non-surgical weight loss and weight stabilization (similar to K99 phase). Participants will be randomized to one of 2 arms (n=72 total; n=36/arm): 1) TRE (selected from the K99 phase) or 2) baseline advice control (CON). Both the TRE and CON groups will be provided with baseline written materials and strategies that promote successful WLM. Participants in the TRE group will be instructed to self-monitor dietary intake and body weight in a smartphone application and receive behavioral counseling. Measurements will be obtained at baseline (0), 3, 6, and 12 months. The primary outcome is body weight regain at 12 months. The candidate will explore the effects of randomization assignment and WLM on appetite biomarkers and subjective appetite via a visual analog scale at 6 mos and 12 mos (Exploratory Aim). Ultimately, this will establish the candidate as an independent investigator conducting interdisciplinary bio-behavioral research using lifestyle interventions to become a leader in WLM.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY – OVERALL Xenotransplantation has the potential to provide nearly unlimited organs for transplantation. Extension of xenograft survival in nonhuman primates (NHP) and xenotransplants to living and brain-dead humans have advanced the field toward clinical trials. Despite these advances, xenograft survival in the few clinical cases performed to date have been limited by infectious complications (of both host and donor origin) and by the development of cellular and antibody-mediated rejection. The development of sensitive assays to detect host and donor microbes, coupled with platforms to guide clinical responses, and assays to detect early markers of xenograft rejection would improve outcomes of clinical xenotransplants. At NYU Langone, we developed an innovative decedent model to study the results of xenografts transplanted to humans and gain experience in a fail-safe environment that also enables frequent sampling of recipient blood and tissues. We have now performed three kidney and two heart xenotransplants in the decedent model, along with one clinical kidney xenotransplant. We have performed innovative multi-omic analyses of the recipients of these transplants. Our preliminary data show that we can detect both host and donor-derived microbes in decedents with an unbiased approach, and that we can detect donor-derived cell-free DNA in recipients at the time of biopsy- proven rejection episodes. We now plan to biobank specimens from 21 additional decedents and perform multi-omic analyses of these recipients. Our program consists of two projects: Project 1 develop sensitive assays to detect host- and donor-derived microbes post-transplant and develop platforms that guide effective clinical management of infections in xenograft recipients; Project 2 will develop assays to detect donor-derived cell-free DNA in recipient blood. We will expand these studies by developing a methylation atlas of pig cells that will enable us to identify the cell of origin of any donor-derived cell-free DNA we detect. The projects will be supported by two Cores: The Molecular Science core contains biobanked specimens from the 5 decedents we have performed to date. We will biobank specimens from the additional 21 decedents and then perform multi-omic analyses of these samples. The Core will also support data analysis of these results. The Administrative Core will provide administrative oversight, coordinate internal scientific meetings, organize travel to consortia meetings, and ensure each Project is meeting their defined milestones. Collectively, these projects will provide a new level of understanding of the management of xenograft recipients and the human immune response to xenografts. Upon successful completion of the proposed research, we expect to develop platforms that improve the clinical management of patients who enroll in the first clinical trials of xenotransplantation, with the long-term goal of developing xenotransplantation into a standard clinical treatment option for organ failure.

NIH Research Projects · FY 2025 · 2025-07

Project Abstract Over two decades after the 9/11 terrorist attacks, many exposed community members in the World Trade Center (WTC) Environmental Health Center (EHC) still struggle with ongoing airway symptoms, with functional abnormalities in large and/or small airways as indicated by spirometry measures of airflow and/or oscillometry measures of airway heterogeneity. Recent studies within WTC cohorts are drawing attention to the complex interplay between WTC exposures and Body Mass Index (BMI) on lung function trajectories. Notably, the WTC Fire Department of the City of New York (FDNY) cohort has identified BMI as the most significant metabolic syndrome characteristic in the risk of developing WTC-related lung diseases. However, there remains a significant gap in understanding this relationship within the WTC Survivor population, which could significantly benefit from weight and BMI management to improve lung function and physical health. Additionally, our unique access to oscillometry data, reflective of small airway function, provides a novel opportunity to assess changes in lung function that may not be captured in large airway spirometry measures. Moreover, the demographic diversity of the WTC Survivor population highlights an essential opportunity to determine the health disparities within the WTC EHC and the critical need to address them. In this project, we propose to tackle the important scientific questions related to BMI, lung function trajectories, and health disparities among WTC Survivors, through the following specific aims: Aim 1: Assess the complex interplay between WTC exposures and BMI on lung function trajectories in WTC Survivors, with a unique focus on small airway oscillometry data. Aim 2: Utilize advanced analytics of algorithmic fairness tools and machine learning to identify factors driving lung function trajectory disparities in WTC Survivors, particularly for BMI and related key factors. Upon its completion, the proposed project will improve our understanding of how BMI influences the health effects of severe respiratory diseases related to the 9/11 terrorist attacks and help develop more effective prevention and treatment interventions that will shape equitable health policies.

NIH Research Projects · FY 2025 · 2025-07

The collapse of the World Trade Center on September 11, 2001 resulted in the massive release of dust, gas, and fumes with potential exposure to known and suspected carcinogens for thousands of individuals. Historically, most of the research focused on first-responders. However, the impact of the WTC exposures on general population, particularly on the health of younger people and women remains poorly understood. The current application will focus on WTC-exposed non- responder women who were diagnosed with breast cancer. The epigenome acts as an interface between the genome and the environment. It is plastic, changing with environmental exposures, thereby regulating transcription. We have previously demonstrated substantial methylation differences in peripheral blood between WTC-exposed and unexposed cancer-free women. However, it is unknown whether blood-based differences correspond to changes at the breast tissue level. The overall objective of this project is to improve understanding of how WTC exposure contributed towards breast cancer development at the molecular level. The project addresses the hypothesis that WTC-exposed women with breast cancer may have tissue- specific epigenetic and transcriptomic profiles that are different from those in unexposed breast cancer patients. To test the study hypothesis, we will compare breast tissue samples of WTC-exposed and unexposed women diagnosed with breast cancer using advanced Illumina Infinium MethylationEpic arrays for global methylation analyses and Nanostring GeoMx Digital Spatial Profiler for whole transcriptome analyses. The project addresses the knowledge gap regarding breast tissue-specific epigenetic and transcriptomic changes in understudied population of WTC-exposed women survivors. Our results could lead to identification of consistent functional pathways and novel tissue-specific biomarkers that will enhance breast cancer diagnosis and treatment in this understudied population.

NIH Research Projects · FY 2026 · 2025-07

Opioid use disorder (OUD) is a leading cause of morbidity and mortality across the U.S, with wide ranging effects across population subgroups. Medications for opioid use disorder (MOUD) substantially improve health and reduce overdose risk, but engagement and retention in MOUD remains low, with the most high-risk patients cycling in and out of care. Hospital systems frequently interact with high-risk patients with OUD, and are increasingly piloting innovations to improve OUD treatment engagement. However, little is known about the influence of such initiatives on long-term patient care trajectories and outcomes, or how they impact different types of patient subgroups. Studying these trajectories is critical but challenging as disjointed healthcare data complicate tracking outcomes of individuals who move across or outside health systems. Moreover, traditional epidemiologic methods are not set up to characterize complex, heterogenous, and non-linear patterns of OUD care trajectories across multiple timepoints and settings. The current proposal aims to apply novel research approaches to better characterize real-world OUD treatment trajectories following opioid-related hospital encounters, how they differ across patient subgroups, and measure their relationship to hospitalization, overdose and mortality outcomes. To do this, we will focus on patients who experience opioid-related hospital encounters between 2021-2024 at any NYC hospital, with an in-depth focus on patients receiving care at Health + Hospitals, the largest safety-net health system in the U.S. We will partner with public NYC agencies to link multiple administrative databases, including all-payer hospital discharge records, electronic health records, Medicaid claims, neighborhood census data, and mortality records at the patient-level. We will then apply State Sequence Analysis - a novel methodology that uses machine learning to identify clusters of patients that share similar trajectories over time - to: (1) Characterize real-world trajectories of OUD treatment in the year following hospital encounters among treatment-naïve individuals, and determine how care pathways differ based on hospital-centered interventions to initiate MOUD; (2) Assess how OUD care trajectories differ across different patient subgroups, using individual and neighborhood characteristics to identify gaps in continuity of care; and (3) Examine relationships between longitudinal OUD treatment trajectories and subsequent hospital encounters, overdose, and mortality. This methodologically and conceptually innovative proposal will advance knowledge on patterns and gaps in OUD treatment among a high-risk population with poor access to care, and generate evidence on the influence of hospital interventions and treatment modalities on hospitalization and mortality. The unique collaboration between academic researchers and health system and government leaders will ensure translation of findings to actionable treatment and policy interventions, thus advancing NIDA’s goal to improve delivery of high quality, effective services for prevention and treatment of substance use disorders and related outcomes.

NIH Research Projects · FY 2025 · 2025-07

Lower Baseline-FEV1 (cross sectional forced expiratory volume at one second at the beginning of longitudinal follow-up) and FEV1-slope (decline in FEV1 over longitudinal follow-up) are mortality risk factors in FDNY World Trade Center exposed rescue and recovery workers. During our investigation of FEV1-slope and mortality, we observed increasing FEV1 visit-to-visit variability is a previously unreported mortality risk factor. We define FEV1 visit-to-visit variability as the standard error of the FEV1-slope modeled by linear regression. Each doubling of FEV1 visit-to-visit variability increased mortality hazard by 22% in the FDNY cohort (Hazard ratio (HR) 1.22, 95% confidence interval (CI): 1.13-1.33 p<.001). Similarly, in an independent community-based replication cohort, doubling FEV1 visit-to-visit variability increased mortality hazard by 17% (HR 1.17, 95% CI:1.14-1.20 p<.001). Inhaled corticosteroids and long-acting beta agonists (ICS/LABA) treated lung injury in 2,209/11,745, 18.8% of the cohort. A feasibility study of 284/2,209,12.5% of those treated for more than two years, assessed if FEV1 visit-to-visit variability is useful as a treatment outcome measure after ICS/LABA initiation. We define response to ICS/LABA as a decrease in FEV1 visit-to-visit variability after treatment initiation (ratio pre/post FEV1 visit-to-visit variability >1). Forty percent (113/284) of the patients in the feasibility study responded to ICS/LABA treatment. Compared with treatment non-responders, responders had a trend to 91% lower mortality odds (OR 0.09, 95%CI. 0.01-1.40 p=0.09). These data are consistent with this R21’s hypothesis: FEV1 visit-to-visit variability is a mortality risk factor and ICS/LABA can improve FEV1 visit-to-visit variability. When compared to ICS/LABA non-responders, ICS/LABA responders, who reduce visit-to-visit variability after ICS/LABA initiation, will have lower mortality. AIM1 will assess the sensitivity and specificity of mortality prediction models that already incorporate well-accepted risk factors of baseline-FEV1 and FEV1- slope to test the hypothesis that adding visit-to-visit FEV1 variability improves their sensitivity and specificity. Aim 2, will use patients as their own controls to determine if ICS/LABA treatment reduces FEV1 visit-to-visit variability and will assess if response to ICS/LABA treatment is associated with reduced mortality. If the data developed in this R21 demonstrates that FEV1 visit-to-visit variability is a mortality risk factor and that treatment response is associated with lower mortality after controlling for well-accepted risk factors including baseline- FEV1 and FEV1-slope, then those with elevated FEV1 visit-to-visit variability who have not been treated could have pulmonary evaluation for consideration of ICS/LABA treatment. This grant’s focus on FEV1 visit-to-visit variability is novel and critically important because it may predict mortality and assess treatments to modify outcomes early in the disease course when treatment is more effective.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Alphaviruses are mosquito-transmitted human pathogens capable of explosive outbreaks and devastating disease. Alphaviruses cause a viremic infection where virus can be disseminated throughout the organism. Microvascular endothelial cells play critical roles in vascular homeostasis and protecting the vasculature from invading pathogens. How alphaviruses infect microvascular endothelial cells and how these cells respond to infection is a major knowledge gap. Filing the gap of viral and cellular determinants of endothelial cell infection would provide novel targets to block disseminated infection. In preliminary studies to understand chikungunya virus (CHIKV) infection in primary human cardiac microvascular endothelial cells (hCMECs), we found that the Indian Ocean Lineage of CHIKV (CHIKV-IOL) was completely restricted in hCMECs, while Asian lineages of CHIKV and other alphaviruses were not. When we addressed where in the viral life cycle CHIKV was restriction, we found CHIKV-IOL to be blocked at viral entry and egress, and not genome replication. These exciting preliminary data allow us to hypothesize that there are both viral and host determinants that restrict CHIKV-IOL in microvascular endothelial cells at multiple steps in the life cycle. We will test this hypothesis using primary human microvascular endothelial cells and identifying the CHIKV-specific viral factors that contribute to hCMEC infection (Aim 1) and investigating hCMEC cellular factors and pathways that restrict CHIKV-IOL (Aim 2). Understanding how pathogenic alphaviruses are restricted in primary human cells will identify viral and host pathways we can target to block virus infection, spread, and disease.

NIH Research Projects · FY 2025 · 2025-07

SUMMARY The World Trade Center (WTC) attacks on September 11, 2001, exposed first-responders to toxic particulate matter (PM) resulting in morbidity and disability. Accelerated aging due to environmental and nutritional exposure is an area of investigation of global significance and pertinent to our WTC-exposed cohorts. We have identified that metabolic syndrome was a risk factor of worsening lung disease, and implemented a successful multidisciplinary dietary randomized clinical trial; Food Intake REstriction for Health OUtcome Support and Education (FIREHOUSE). Subjects that lost weight had improved lung function and respiratory quality of life symptoms. We have assayed their gut microbiome, metabolomics, and inflammatory profile, and collected genetic samples. However, the mechanisms that link WTC-PM exposure, metabolism, aging, hyper-inflammation and WTC-PM related lung disease are yet to be identified. We will focus on innovative concepts linking aging and increased autoantibodies under the hypothesis that both processes can cause persistent hyper-inflammatory states contributing to worsening lung function. We further hypothesize that: i. A nutritional lifestyle intervention can modify aging and autoantibody biosignatures in a WTC-PM exposed cohort. ii. WTC-PM exposure induced chronic inflammation and metabolic markers will be exacerbated by metabolic syndrome, age and transient depletion of a key anti-inflammatory mediator in mice, increasing autoantibodies and decreasing lung function. AIM 1. Lifestyle, Aging and Autoimmunity. Defining the MultiOme of Aging and Autoimmunity in the context of a nutritional intervention is designed to identify a subgroup of firefighters who have a chronic hyperinflammatory state, accelerated aging, and increased circulating autoantibodies that will be associated with health and lung function metrics. We will then integrate the MultiOME using systems biology to optimize disease specific models and identify mechanistic processes of WTC-Lung Injury. AIM 2. Mechanistic pathways will be examined utilizing a mouse model to determine a cause-effect relationship of WTC-PM exposure and high fat diet on aging and the exacerbation of hyperinflammatory responses (including autoantibodies). We will use mice from a well-established, commercial model of aging with features of metabolic syndrome, and expose them to WTC-PM. We will further study the effects of blocking a key anti-inflammatory cytokine to determine adverse effects (lung function, structural lung and heart remodeing) of WTC-PM exposure. The overall goal of this 2-year pilot is to begin to understand the mechanisms by which our nutritional intervention improves health outcomes, focusing on the interactions between a diet high in fat, aging, diminished control of inflammation and autoimmune antibodies. Our study outcomes will improve the overall understanding of how WTC lung disease persists, enhance our ability to better manage adverse health effects, benefit the health/well- being of WTC-exposed first-responders and falls squarely within the purview of the James Zadroga 9/11 Health & Compensation Act. This pilot will help lay the ground work for targeted immune/lifestyle driven therapies.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY The overall success of heart transplantation (HTx) as a lifesaving therapy for patients with end-stage heart failure is attributable to the development and refinement of the current immunosuppressive drug regimens. However, these agents carry well-known risks for malignancies, new onset diabetes, chronic kidney disease, and other conditions that limit quality of life and long-term survival. The ability to titrate immunosuppression (IS) to precisely match an individual recipient’s need would prevent both over-IS with its associated toxicities, and under-IS with its risks of graft rejection. We propose to investigate the role of HLA eplet mismatches (EMM) between donors and recipients as a biomarker for immunologic risk in HTx patients. In kidney transplantation (KT), EMM analysis has been validated as such a biomarker, and established EMM thresholds defining high and low risk categories are being used to guide IS. Since high-resolution HLA genotyping, required to calculate EMM, has not historically been performed, the ideal prospective study of EMM in HTx would require long-term follow-up and thus not yield meaningful results for several decades. To answer the question now, as to whether EMM is a valuable biomarker for immunologic risk in HTx, we propose the use of three patient cohorts which will provide complementary outcomes data with which to model associations between EMM and immunoreactivity. In Aim 1 we will use a national dataset to impute high-resolution HLA genotyping based on the low-resolution HLA typing that is captured in the registry. By leveraging population-specific high-resolution haplotype frequency data and reported ethnicity, we will calculate imputed EMM for a large cohort (N=51,530) of HTx recipients. Test imputations demonstrate good calibration when verified against known high-resolution typing. We will evaluate the association of EMM with rejection, re-transplantation, and mortality captured in the database. Using machine learning we will evaluate the EMM risk thresholds defined in KT patients and explore ways to improve them for HTx recipients. In Aim 2 we will assess the association of EMM with cardiac allograft vasculopathy (CAV), and with biopsy-proven rejection using a protocol HTx biopsy dataset. In Aim 3 we will study a single-center observational cohort to examine the association of EMM with early indicators of immunoreactivity in HTx recipients, including the development of de novo donor specific antibody and the presence of donor-derived cell free DNA. There is an urgent need for a valid biomarker to gauge immunologic risk in HTx recipients. Our use of existing and prospectively collected data from complementary patient cohorts will enable us to evaluate EMM as a biomarker for HLA mismatch driven immunoreactivity. We aim to demonstrate that EMM can be utilized to accurately and risk-stratify patients at the time of HTx. This work will establish a foundation for future clinical trials that can fine-tune IS management, reducing the preventable morbidity and mortality that accompanies both over- and under-IS.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY Staphylococcus aureus (S. aureus), a leading global pathogen, is the primary microorganism implicated in spine infection. Because patients’ backpain symptoms are often misdiagnosed, paralysis may ensue in up to 75% of patients, and up to one-third of them may die from the disease. Despite recognition of the severe disease morbidity and mortality, the molecular mechanism of human spine infection remains undefined. S. aureus-human coevolution likely favor selection of isolates that are adept at virulence factor regulation adapt to the human host. One such adaptation is the ability to interfere with the C5a-hC5aR1 axis, a powerful component of the human innate immune system, through the production of three human-specific virulence factors, the Panton-Valentine Leukocidin (LukSF-PV/PL), γ-hemolysin CB (HlgCB), and the Chemotaxis Inhibitory Protein of S. aureus (CHIPS). These toxins target hC5aR1 to prevent C5a binding, thus disrupting neutrophil chemotaxis and function, enabling pathogen evasion of host killing. Up until now, species- specific receptor limitations have hindered accurate modeling of the effects of these toxins. However, the development of a novel humanized mouse model with knock-in hC5R1 (hC5aR1KI) set the stage for us to evaluate their contributions in invasive S. aureus infection. When we evaluated S. aureus bacteremia in wild-type and hC5aR1KI mice, we observed that hC5aR1KI mice developed highly penetrant mobility deficits involving the hindlimbs owing to infection of the spine, an important complication of S. aureus bacteremia in humans. We further dissected the contribution of each hC5aR1 targeting virulence factor, as well as examined the contribution of other S. aureus virulence factors, and implicated CHIPS and at least one of the 21 sortase A dependent cell wall-anchored surface proteins (SADCASP) as critical contributors to spine infection. To evaluate the roles of CHIPS and SADCASP in human spine infection, I will: 1) Clarify the role of surface proteins in spine infection, and prepare to test my hypothesis that immune interference and immune evasion properties of SADCASP and CHIPS contribute to their success in spine infection. 2) Determine the mechanism of injury in S. aureus spine infection with respect to CHIPS and SADCASP, and evaluate how CHIPS’ interference with C5a-hC5aR1 and SADCASP’s immune evasion properties contribute to primary and secondary spinal cord injury. With a better understanding of how CHIPS and SADCASP promote spine infection, I will 3) Explore vaccination approaches targeting CHIPS. Using a novel and clinically relevant mouse model of S. aureus infection, I hope to better understand the mechanisms behind how CHIPS and the SADCASP empower S. aureus to cause spine infection in humans, and use this information to identify potential vaccination strategies to prevent its debilitating impact.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY The immune system relies on a carefully orchestrated network of spatially organized compartments to function effectively. These architectural ‘hubs’ exist in the form of secondary and tertiary lymphoid organs. Immune cells traffic through immune organs via the lymphatic vasculature, a network of vessels which, in addition to providing a transport route for antigens, cells, and metabolites, also plays active roles in regulating host immunity. Our published and preliminary work establishes the lymphatic vasculature in the skin as a necessary route for both immune activation (ON) and immune resolution (OFF) signals. Further, we have demonstrated that dermal lymphatic vessels remodel their inter-endothelial junctions (termed ‘zippering’) with functional consequences for pathogen dissemination and immune induction. Our preliminary data now indicates that infection-induced lymphatic remodeling tunes the germinal center response and thereby facilitates the generation of antibody- dependent protective immunity. What regulates perifollicular lymphatic growth in the context of inflammation and whether lymphatic vessels play an architectural role in dictating germinal center area remains completely is unknown. In this proposal we aim to test the hypothesis that peri-follicular lymphangiogenesis shapes germinal center responses by constraining GC size and thereby optimizing the search space for GC B cell selection and antibody output. We will test this hypothesis in two aims: (1) we will elucidate the mechanisms governing lymphangiogenesis in acute and homeostatic inflammation in different tissue microenvironments; (2) determine the functional consequences of lymphatic encapsulation of B cell follicles on germinal center dynamics. To complete these aims, we couple genetic tools with robust immunological assays and high-resolution imaging to resolve the systems-level interactions between lymphoid organ organization, germinal centers, and antibody responses in vivo as a function of the connecting lymphatic vasculature. We propose that lymphatic vessel transport is a poorly understood but active determinant of germinal center fitness and that identification of specific molecular mechanisms that regulate their function provides novel therapeutic opportunities to direct the quality of antibody responses. Our work will not only develop our physiological understanding of lymphatic transport and its contribution to immunity but further suggest novel design principles for how to instruct optimal immunological reactions during vaccination, infections, or autoimmunity. In this way our work may help to inform clinical vaccine development and immunotherapy and may identify targets to maintain tolerance during homeostasis.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT KRAS is among the most frequently mutated genes in human cancer with most mutations affecting codon 12. KRASG12C (G12C) is especially common in non-small cell lung cancer (NSCLC), comprising ~half of all KRAS mutations in this disease (~15% of NSCLC overall). Approximately 40% of colorectal carcinoma (CRC) cases are KRAS-mutated (~10% G12C), while ~1-2% of pancreatic ductal adenocarcinomas (PDAC) have this mutation. Mutant KRAS had long been viewed as “undruggable”, until pioneering chemical biology and medicinal chemistry studies resulted first in G12C-specific inhibitors (G12Cis), and later in a plethora of agents targeting other mutant alleles or even wild type (WT) KRAS and most/all mutants (“pan-KRAS”). Thus far, two small- molecule G12Cis, sotorasib and adagrasib, have been FDA-approved as second-line agents for NSCLC. While the magnitude of the conceptual advance provided by G12Cis and other RAS inhibitors (RASi) cannot be overstated, the clinical response to single-agent G12Cis has been modest. Specific co-mutations (e.g., KEAP1) can impair the response of G12C-mutated NSCLC, while as single agents, G12Cis have minimal activity in G12C-mutated CRC. Even when tumors initially respond, resistance emerges rapidly via multiple mechanisms. Early results suggest that intrinsic and/or emergent resistance will also limit the clinical impact of other RASis. Whether the same resistance mechanisms will plague other KRAS inhibitors remains unclear, yet what does seem likely is that durable responses and, ultimately cures, of KRAS-mutated NSCLC will require drug combinations, potentially including conventional chemoradiation and immune therapies. Rapidly and efficiently devising such strategies would address a major unmet medical need. We recently reported the results of a genome-wide CRISPR/Cas9 “dropout” screen for adagrasib synthetic lethal (SL) genes in four (4) KRASG12C;STK11-co-mutated cell NSCLC lines, three (3) of which are also KEAP1-mutated. We identified ~40 genes that were SL in at least 3 lines, including three (3) serine/threonine kinases (STKs), and ~330 that were SL in at least two lines. Although our screens focused on SL (i.e., dropout) genes, we also identified 30 enriched “resistance hits” in at least 2 lines. Functional studies provided initial validation of the three STKs, as well as ROCK1,2 (which emerged from studies of other recurrent SL genes), but more complete evaluation, including mechanistic and in vivo efficacy studies, was lacking, as was the applicability of screen “hits” to other G12C malignancies and other RASis. We propose to tackle these key gaps via a combined biochemical, genetic, and functional genetic approach. Specfically, we will 1) evaluate four STK family members as SL targets more broadly and in vivo and clarify the mechanistic basis of synthetic lethality, and 2) use CRISPR/Cas9 “mini-libraries” to define the synthetic lethality/resistance landscape for G12Cis and other RASis. Our results will address a critical unmet need in medical oncology by credentialling new targets for RASi combinations.