Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2026 · 2023-08

In the United States, more than a third of patients are referred to a specialist each year, and specialist visits constitute more than half of outpatient visits. Even though all physicians highly value communication between primary care providers (PCPs) and specialists, both PCPs and specialists cite the lack of effective information transfer as one of the most significant problems in the referral process. Therefore, it is critical to investigate a new method to improve communication during care transitions. With their ubiquitous use, it is recognized that electronic health records (EHRs) should ensure a seamless flow of information across healthcare systems to improve the referral process. But, a lack of accessible and relevant information in the referral process remains a pressing problem. Recently, emerging deep learning (DL) and natural language processing (NLP) methods have been successfully applied in extracting pertinent information from EHRs and generating text summarization to improve care quality and patient outcomes. However, existing technologies cannot be applied to process heterogeneous data from EHRs and create high-quality clinical summaries for communicating a reason for referral. Responding to PA-20-185, this project will develop and validate a novel informatics framework to collect and synthesize longitudinal, multimodal EHR data for automatic referral form generation and summarization. While the referring provider and specialist can be any type of provider for any condition, the focus in this application has been on headache for primary care, because it is an extremely common symptom and affects people of all ages, races, and socioeconomic statuses. More importantly, relevant information needed for headache referrals has been defined in local and national evidence-based practice guidelines. Therefore, a health information technology solution to make these data accessible will empower communication between PCPs and specialists, which can improve the care of millions of patients suffering from disabling headache disorders. Based on our preliminary data and our experience with an interdisciplinary team of data scientists and physicians, we plan to execute specific aims: 1) Convert text-based guidelines into a standards-based algorithm for electronic implementation; 2) develop models to automatically populate data from EHR and clinical notes to fill the referral form; 3) create a framework to summarize the longitudinal clinical notes to fill out the referral form; and 4) develop and validate the headache referral system with a user-centered design approach. The research proposed in this project is novel and innovative because it will produce and rigorously test new solutions to improve the communication between health professonals to ensure that safe, high-quality care is provided and care continuity is maintained. The success of this project will (1) fill important gaps in our knowledge of understanding the types of information exchange that will optimize patient care during transitions and (2) provide evidence-based solutions to enable the exchange.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY Stroke is a devastating disease and leading cause of death and disability in the United States. Ischemic stroke results in massive activation of numerous immune cells that can infiltrate the brain following blood-brain-barrier breakdown. The gut microbiota has previously been identified as a significant factor affecting outcome and severity of ischemic stroke in clinical studies and animal models. However, mechanisms underlying the modulatory role of microbiota on immune cells following stroke remain unclear. Dendritic cells (DCs) act as the bridge between innate and adaptive immunity, with their ability to sample material from the intestinal lumen and shape T-cell responses. Antibiotic-induced alteration of microbiota in mice results in stroke neuroprotection in mice following middle cerebral artery occlusion model of ischemic stroke compared to control mice carrying conventional microbiota, which are similarly treated but carry antibiotic-resistant microbiota resulting in microbiota similar to that of naïve mice. This effect is attributed to the greater capacity of intestinal and mesenteric lymph node dendritic cells of mice carrying “altered” microbiota to induce T-regulatory cells (Tregs) in the small intestine which subsequently suppress destructive pro-inflammatory IL-17+ γδ T cells that traffic to the brain following stroke. Using our in vitro model to simulate intestinal DC-T cell interactions, we show that priming naïve DCs with isolated contents from the small intestine (SIC) of mice carrying “altered” or “conventional” microbiota and subsequent co-culture with CD4 cells similarly induces greater proportions of Tregs following SIC from mice carrying “altered” microbiota compared to SIC from mice with “conventional” microbiota. This proposal seeks to elucidate the mechanism by which altering microbiota may result in changes in pattern-recognition receptors or toll-like receptor ligands that are responsible for a DC-tolerizing phenotype and Treg induction observed with microbiota alteration in mice. Using a variety of in vitro and in vivo approaches, I aim to identify the DC receptors and signaling machinery responsible for sensing these luminal contents and producing a tolerogenic phenotype, determine DC-produced signals/cytokines necessary for intestinal Treg induction, and establish the role of pro- inflammatory IL-6 in stroke neuroprotection vs poor stroke outcome in mice carrying “altered” or “conventional” microbiota. In summary, I seek to understand how intestinal DC receptor ligands that are microbiota-dependent can act as regulators of intestinal immunity and stroke outcome, as well as identify potential therapeutic targets.

NIH Research Projects · FY 2025 · 2023-08

1 2 functions, 3 have 4 spatial resolution and sensitivity. Such specifications impact both neurologic and neuro-oncologic diseases. In 5 the former, they allow detecting, quantitating, and tracking small changes of PET signal in minute brain regions 6 such as brain nuclei that have been implicated in many neurologic diseases. In the latter, they improve the 7 accuracy of tumor target volume definition in radiotherapy and surgical resection, thus treatment outcome. 8 The only commercial brain-dedicated PET is the HRRT, a 2 decade old technology that has been discontinued. 9 Therefore, there is compelling need to develop the next generation brain-dedicated PET with ultra-high 10 specifications to improve diagnostics that can institute therapies earlier in the evolution of the disease. 11 This proposal brings together two highly collaborative teams from Weill Cornell Medicine (WCM) and the 12 Institute for Instrumentation in Molecular Imaging (i3M), with an industrial partner, Oncovision, to build an 13 ultra-high performance brain-dedicated PET, UHB-PET. UHB-PET will exhibit: (i) volumetric spatial 14 resolution of ~0.5mm3 across the gantry, that is >4x better than that of the best brain-dedicated PET being 15 developed; (j) effective sensitivity >26x that of the brain PET with the highest spatial resolution being developed. 16 Our intensive experimental and Monte Carlo simulation results prove that our goals are highly achievable, 17 which we will attain as follows: Specific 18 maximize 19 FOV 20 and 21 Quantitative machine 22 learning to accurately determine the 3D position of 511keV 's interaction within the semi-monolithic slab, infer the 23 attenuation-corrected PET without CT scans, and minimize image noise, thus reduce the administered dose), (d) 24 use a preconditioned fixed-point image reconstruction approach to suppress the noise in sub-millimeter size 25 pixels, (e) adopt motion tracking tool we previously developed to correct for inter- and intra- head motion during 26 dynamic PET imaging, and (f) adopt methods from our previous work to accurately image-derive the input function 27 for kinetic modeling. Specific Aim 3: Assessment of image noise, target lesion visibility and quantitative accuracy 28 attained by the scanner in a characteristic set of specific neurology and neuro-oncology human studies. 29 The ultimate goal is a fully operational ultra-high performance dedicated brain PET scanner with accurate 30 quantitative capabilities for diagnosing and monitoring treatment in brain diseases. . Positron Emission Tomography (PET) is a powerful quantitative tool for studying metabolic and biochemical pharmacology, and pathology in living brains. In the past 3 decades, a myriad of brain PET tracers, been developed.In parallel, PET underwent dramatic advancement in technology that enabled much higher Aim 1 : In 2.5 years we will build UHB-PET with trapezoidal-shape (to sensitivity) semi-monolithic LYSO slabs coupled to high-performance SiPM readout, 26.7cm axial and ~28cm diameter, high 200psec detector timing resolution, isotropic spatial resolution <0.8mm FWHM 0.72mm Depth-of-interaction (DOI) resolution. Specific AIM2 : In parallel with AIM1, we will develop a Image Reconstruction tool. We will (a) incorporate accurate physics modeling, (b) use

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY / ABSTRACT Suicide rates are high in late adulthood, peaking in midlife among women and late life among men. Social disconnection also peaks in late life and increases risks of suicide, persistent suicidality, and poor response to psychosocial interventions. Our proposal responds to RFA-MH-22-135, outlining the critical need to identify the mechanisms mediating the relationship between social disconnection and late-life suicidality and develop efficacious interventions to target these mechanisms. The proposed study leverages methodological and conceptual innovations, developed by our team, to investigate target engagement of the Positive Valence System (PVS) during a novel social reward psychotherapy for mid- and late-life suicidality. We designed Engage & Connect, a remotely delivered psychotherapy that targets social disconnection by increasing engagement in rewarding social activities. Engage & Connect aims to alter disturbances of the PVS that may underlie late-life suicidality. In this study, 128 adults aged 50-80 with major depressive disorder and suicidal ideation will be randomized to 9 weeks of Engage & Connect or Symptom Review and Psychoeducation (SRP) active control condition. We will measure PVS functions on brain and behavioral levels, through resting state functional connectivity of the PVS and behavioral changes in social reward responsivity using our novel validated STAR task (Social Task for Assessment of Reward). We will employ a cutting-edge “precision imaging” approach to estimate the functional brain map of each individual and track the longitudinal effects of treatment on PVS circuitry. Our rigorous methodological approach will allow us to test, at the individual level, the brain-based and behavioral mechanisms underlying response to psychotherapy that targets social disconnection. Identification of individual patients’ biological and behavioral profiles, linked with treatment response, can guide future psychotherapy personalization and increase its efficacy and reach to vulnerable older adults at risk of suicide.

NIH Research Projects · FY 2025 · 2023-07

PROJECT ABSTRACT Tumor necrosis factor (TNF) is a pleiotropic cytokine that promotes host defense, cell survival and tissue regeneration under homeostatic condition However, if dysregulated and overexpressed, TNF is a major driver of chronic inflammation. Excessive TNF production in the gastrointestinal tract targets the epithelium, drives increased cell death, and is sufficient to elicit substantial tissue inflammation and chronic disease. Blockade of TNF is a widely utilized biologic that provides therapeutic benefit in a subset of inflammatory bowel disease (IBD) patients. Despite this knowledge, the mechanisms that control the beneficial versus detrimental roles of TNF in the intestine are poorly defined. The fundamental focus of this proposal is to mechanistically define a novel pathway that protects the intestine from TNF-driven damage and inflammation. In recently published and new preliminary data, we have determined that group 3 innate lymphoid cells (ILC3s) are essential to protect the intestinal epithelium from TNF-driven damage and inflammation. Surprisingly, this did not occur via traditional effector pathways, and rather involved production of prostaglandins and growth factors. These data provoke a fundamental hypothesis that ILC3s are essential to shape the protective versus pathologic roles of TNF in the intestine, and this balance is disrupted in human IBD where ILC3s are known to become dysregulated. We will mechanistically test this hypothesis by asking the following specific questions: (1) How does ILC3 production of sensing of prostaglandins impact intestinal health and inflammation? And (2) What are the cellular and molecular mechanisms by which ILC3-derived HB-EGF augments intestinal immunity and protects the gut from TNF? Finally, we will directly translate our findings from basic mouse models into samples from IBD patients. Results from these experiments will pave the way for a greater understanding of TNF-driven intestinal damage and inflammation, which could provoke novel preventative, therapeutic or curative strategies for multiple chronic inflammatory diseases.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Candidate: Dr. Kathleen Walsh is an Internal Medicine-trained physician scientist who has spent the past seven years conducting research in Haiti. She has gained first-hand experience in treating drug-resistant TB, learned Haitian Kreyol and trained a research team of Haitian physicians, nurses and lab technicians in the conduct of research in tuberculosis. She has authored 22 publications including 9 first author papers. She has preliminary data suggesting there is an emerging epidemic of isoniazid-resistant tuberculosis (INHr-TB), that adolescents are a sentinel population for INHr-TB in Haiti and that INHr-TB is predominantly being transmitted outside the household in community settings. She seeks to investigate the epidemiology of INHr-TB in Haiti. Career Goals: Dr. Walsh's goals are: 1. To gain skills in molecular epidemiology through utilization of whole genome sequencing to identify molecular disease clustering and INHr-TB transmission. 2. To gain expertise in geographic information systems (GIS) mapping of INHr- and DS-TB in Haiti 3. To acquire skills in biostatistics, specifically analysis of temporal trends and Poisson regression Career Development Plan: Dr. Walsh will have mentorship from Dr. Daniel Fitzgerald (clinical TB epidemiology), Dr. Jean W. Pape (TB disease mapping), Dr. Theodore Cohen (molecular epidemiology), Dr. Denis Nash (spatial epidemiology) and Dr. Myung Hee Lee (biostatistics). She will engage in courses and field work related to her proposed research. She will submit an R01 application based on this data. Environment: The proposed research will occur at Weill Cornell Medicine (New York, USA) and GHESKIO (Port-au-Prince, Haiti), which has 40 years of NIH-supported research and training in TB. Research: INHr-TB is the most common form of drug-resistant TB worldwide. Strains of Mtb typically acquire resistance to INH first and then sequentially to other drugs. Therefore, INH resistance is a precursor to multidrug-resistant TB. Since INH resistance is rarely diagnosed, there are large gaps in our understanding of this disease. Dr. Walsh hypothesizes that INHr-TB is an emerging epidemic in Haiti and adolescents are sentinel populations for this emergence. She further postulates that INHr-TB is predominantly transmitted in community settings and not in the household. She will test 5,183 diagnostic Mtb isolates from 2011 – 2021 to characterize temporal trends in INHr-TB prevalence in Haiti and determine if INHr-TB trends in adolescents precede INHr-TB trends in adults (Aim 1). She will prospectively investigate the conventional, molecular and spatial epidemiology of 360 INHr-TB and 360 DS-TB patients in Haiti and identify congregate settings where INHr-TB transmission is occurring (Aim 2). This project will provide critical insight into the epidemiology of INHr-TB so that we can improve detection and prevention of INHr-TB in Haiti and worldwide.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Most bladder cancer (BC) patients are diagnosed at an early stage. More than 80% of cases are non-muscle invasive BC (NMIBC). The standard treatment involves removing the tumors surgically, followed by intravesical immunotherapy, bacillus Calmette-Guérin (BCG), or intravesical chemotherapy (ITC) to eradicate residual cancer cells. This involves direct instillation of the BCG or drug solution into the bladder via a catheter. However, the cancer recurrence rate is still unacceptably high (50-80%). On the other hand, there is a growing interest in preserving the bladders of muscle invasive BC (MIBC) patients who are ineligible for radical cystectomy with ITC. BCG and ITC have limitations. The treatments are local. The drug solution is unable to reach tumors located in the upper urinary tract. Patients often need to void shortly after drug administration. The catheterization procedure is invasive, which can potentially cause infection and urinary symptoms, resulting in poor patient compliance. Currently, there is also a shortage of BCG. The goal of this project is to develop an approach to counter the significant drug delivery obstacles of BC therapy to improve treatment and survival outcomes. A peptide’s rapid renal clearance can be advantageous for directing treatments to the urinary system (URS). We propose a bio-inert peptide (Bdd) to overcome the drug-delivery barriers. Bdd can be given intravenously rather than intravesically. The use of Bdd as a carrier was shown to promote drugs, such as mertansine (DM1) and doxorubicin (DOX), to be eliminated exclusively via renal clearance, with minimal—if not undetectable— deposition in major organs. We hypothesize that this platform, used as an alternative to ITC, will offer an urgently needed treatment that is more complete and effective. The advantages of such a urinary drug disposing (UDD) system include: (1) continuous drug flow throughout the entire URS, (2) prolongation of bladder-dwelling time (treatment duration), and (3) minimally invasive application. If successful, this approach will also avoid catheterization, improve patient quality of life, and reduce hospitalization costs. Our Specific Aims will focus on preclinical and translational studies to: (Aim 1) investigate the desired physicochemical properties (including functional group, length, and surface charges) and administration parameters (infusion rate and volume) of a newly developed Bdd analogue, Bds, with an improved UDD property, for precision drug delivery to the URS; and (Aim 2) evaluate the therapeutic efficacy and anatomic flexibility of a DM1-Bds conjugate for BC treatment. We will assess DM1-Bds alone or in combination with pembrolizumab, an immune checkpoint inhibitor approved by the Food and Drug Administration, for treating both NMIBC and MIBC. Immune profiling will address the anti- tumor activities. This information will be crucial for significantly improving treatment outcomes. With additional advancements, we also foresee our UDD approach will be unusually transposable and useful for treating other diseases (e.g., bladder infections), simply by replacing the drugs attached to the peptide sequence with antibiotics.

NIH Research Projects · FY 2024 · 2023-07

PROJECT SUMMARY Understanding the role of environmental factors including signals derived from the diet and microbiota is key to improving therapeutic and intervention strategies for gastrointestinal disorders including inflammatory bowel diseases (IBD) and inflammation-associated colorectal cancer (CRC). Dietary fiber can exert immunoregulatory effects through microbial fermentation products including short chain fatty acids (SCFAs). However, the influence of dietary fiber on most microbiota-derived metabolites and their role in immunoregulation remain unclear. In new preliminary studies, I identified that an inulin-rich high fiber diet triggers colonic eosinophilia in a microbiota- dependent manner which exacerbates disease outcomes in murine models of intestinal damage and inflammation-associated CRC. These diet-induced type 2 inflammatory responses are associated with upregulation of microbiota-derived bile acids and activation of mesenchymal stromal cells (MSCs) and group 2 innate lymphoid cells (ILC2s). However, the cellular and molecular components in the host and the metabolic pathways in the microbiota that mediate the dietary effects on intestinal inflammation remain unclear. In Aim 1, I will determine how bile acids regulate type 2 cytokines and eosinophilia during high fiber diet-induced intestinal inflammation. In Aim 2, I will investigate how dietary fiber-induced eosinophils contribute to intestinal damage. In Aim 3, I will employ bacterial genetics and gnotobiotic approaches to identify the microbial metabolic pathways required for high fiber diet-induced intestinal inflammation. Upon successful completion of the proposed aims, I expect to contribute to a fundamentally new understanding of the biology of dietary fiber, microbiota-derived metabolites, and stromal cells in regulating type 2 inflammation which could contribute to rational design of diet- and microbiota-based therapeutic approaches. My overarching career goals are to become an independent investigator and an inspirational mentor at a leading academic institution and to study the mechanisms by which environmental factors regulate intestinal inflammation and gastrointestinal diseases. Completion of my research aims in this proposal will allow me to develop various scientific, professional, and personal skills critical to become a successful independent investigator. These will include acquiring expertise in various fields including metabolomic analyses and genetic engineering with help of my collaborators, as well as developing skills in writing, mentoring, communicating and laboratory management. I will perform the K99 phase in the laboratory of Dr. David Artis, a world leader in the fields of mucosal immunology and host-microbiota interactions. The laboratory has access to all instruments and facilities necessary to complete the proposed aims including a gnotobiotic animal facility and provides an outstanding environment and training program to support postdoctoral researchers. In addition to my mentor and co-mentor, I have support from several distinguished investigators with extensive expertise as advisors and collaborators that will greatly facilitate the completion of my experiments during the K99 phase as I progress to become an independent investigator.

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY Understanding the role of environmental factors including signals derived from the diet and microbiota is key to improving therapeutic and intervention strategies for gastrointestinal disorders including inflammatory bowel diseases (IBD) and inflammation-associated colorectal cancer (CRC). Dietary fiber can exert immunoregulatory effects through microbial fermentation products including short chain fatty acids (SCFAs). However, the influence of dietary fiber on most microbiota-derived metabolites and their role in immunoregulation remain unclear. In new preliminary studies, I identified that an inulin-rich high fiber diet triggers colonic eosinophilia in a microbiota- dependent manner which exacerbates disease outcomes in murine models of intestinal damage and inflammation-associated CRC. These diet-induced type 2 inflammatory responses are associated with upregulation of microbiota-derived bile acids and activation of mesenchymal stromal cells (MSCs) and group 2 innate lymphoid cells (ILC2s). However, the cellular and molecular components in the host and the metabolic pathways in the microbiota that mediate the dietary effects on intestinal inflammation remain unclear. In Aim 1, I will determine how bile acids regulate type 2 cytokines and eosinophilia during high fiber diet-induced intestinal inflammation. In Aim 2, I will investigate how dietary fiber-induced eosinophils contribute to intestinal damage. In Aim 3, I will employ bacterial genetics and gnotobiotic approaches to identify the microbial metabolic pathways required for high fiber diet-induced intestinal inflammation. Upon successful completion of the proposed aims, I expect to contribute to a fundamentally new understanding of the biology of dietary fiber, microbiota-derived metabolites, and stromal cells in regulating type 2 inflammation which could contribute to rational design of diet- and microbiota-based therapeutic approaches. My overarching career goals are to become an independent investigator and an inspirational mentor at a leading academic institution and to study the mechanisms by which environmental factors regulate intestinal inflammation and gastrointestinal diseases. Completion of my research aims in this proposal will allow me to develop various scientific, professional, and personal skills critical to become a successful independent investigator. These will include acquiring expertise in various fields including metabolomic analyses and genetic engineering with help of my collaborators, as well as developing skills in writing, mentoring, communicating and laboratory management. I will perform the K99 phase in the laboratory of Dr. David Artis, a world leader in the fields of mucosal immunology and host-microbiota interactions. The laboratory has access to all instruments and facilities necessary to complete the proposed aims including a gnotobiotic animal facility and provides an outstanding environment and training program to support postdoctoral researchers. In addition to my mentor and co-mentor, I have support from several distinguished investigators with extensive expertise as advisors and collaborators that will greatly facilitate the completion of my experiments during the K99 phase as I progress to become an independent investigator.

NIH Research Projects · FY 2026 · 2023-07

PROJECT ABSTRACT The World Health Organization estimates that 1 in 6 adults over the age of 60 has been a victim of abuse in the past year. Our research demonstrated that nearly one-third of these victims have clinically significant depressive symptoms and 16% have suicidal ideation (Sirey et al., 2015). Tele-PROTECT, is the only manualized, virtual therapy for elder abuse victims experiencing depression. It is offered to victims at the same time that they are receiving services from elder abuse agencies that offer safety planning, support services, and legal services. Tele-PROTECT was designed using a deployment-focused approach with our partners at the New York City Department for the Aging (DFTA). Tele-PROTECT strategies focus on increasing pleasurable and rewarding activities to reduce depression, and goal setting and defining the action steps to increase safety related empowerment. In our randomized controlled pilot project, participants receiving PROTECT had significant decreases in symptoms of depression as compared to those in the referral control condition (Sirey et al., 2021). Our preliminary work during the pandemic, demonstrated the feasibility and effectiveness of virtual delivery of PROTECT via video (called Tele-PROTECT) to a large sample of diverse victims. This project will test the effectiveness of Tele-PROTECT as compared to a video-delivered attention control condition (DepEd). We will recruit 140 English- and Spanish-speaking elder abuse victims from five New York City (NYC) elder abuse agencies. We hypothesize that Tele-PROTECT participants will experience greater decreases in depressive symptoms and greater increase in safety-related empowerment compared to an attention control (DepEd). Elder abuse agency staff will refer depressed victims who agree to research staff. Eligible elder abuse victims who consent, have a baseline assessment and meet inclusion criteria will be randomized to Tele-PROTECT or DepEd. Follow up assessments will be conducted at weeks 3, 6, 9, and 12 after baseline. Trained Research Assistants blinded to study hypotheses will assess depression severity (MADRS) and increased safety-related actions (MOVERS), as well as participation in activities, social isolation, and criminal justice outcomes. In collaboration with a national organization of elder abuse agencies (NAPSA), we will use a mixed methods design with multiple stakeholder groups to collect data on the barriers and facilitators to the implementation of Tele-PROTECT nationally. Tele-PROTECT has demonstrated feasibility, acceptability, and preliminary effectiveness. This project will test a scalable manualized therapy with the potential for national implementation to serve depressed victims of elder abuse.

NIH Research Projects · FY 2025 · 2023-07

Summary Tryptophan (Trp) catabolism is a complex pathway that generates over fifty metabolites in a cell-specific manner. Besides being the precursor of serotonin and melatonin, tryptophan generates a cascade of metabolites known as kynurenines. Kynurenine metabolites are regarded as one of the most powerful mediators of immune regulation. The thrust of this application stems from our original observations that: (i) lymphatic endothelial cells (LEC) and dendritic cells (DCs) secrete a previously unidentified biogenic amine, 3HKA, which derives from a lateral pathway of Trp catabolism, whose function is currently unknown; (ii) 3HKA exhibits a clear anti- inflammatory profile by inhibiting the STAT1/NF-κΒ pathway in both mouse and human dendritic cells (DCs) with a consequent decrease in the release of pro-inflammatory chemokines and cytokines; most notably, IL-6, IL12p70 and TNFα; (iii) in vivo, 3HKA exerts protective effects in the experimental model of psoriasis by decreasing skin thickness, erythema, scaling and fissuring. In a model of nephrotoxic lupus, 3HKA improved proteinuria and serum urea nitrogen, overall ameliorating the immune-mediated glomerulonephritis and renal dysfunction. As such, the ultimate goal of this application is to fully characterize the biological activity of 3HKA. In Aim 1, a series of biochemical and biophysical analyses will be employed to identify the enzyme responsible for 3HKA production and investigate the regulation of its synthesis under physiological and pathological conditions. In Aim 2, by using biotinylated derivatives of 3HKA, and a series of biochemical and biophysical experiments, we will identify the receptor target of 3HKA and its expression profile. In Aim 3, using targeted and untargeted phosphoproteomic approaches, screening of kinase libraries and CRISPR/Cas9 knock down strategies we will characterize the 3HKA signal transduction pathway. Additionally, we will analyze the effect of 3HKA on T cell activation and differentiation into a TH1, TH2, TH17 phenotype, naïve vs effector and memory T cell transition and 3HKA effect on maturation/activation of different APC (DC, pDC, MΦ). Immunophenotyping will be performed both in vitro and in vivo, using psoriasis or nephrotoxic lupus as animal models. Finally, in Aim 4 we leveraged our knowledge on 3HKA and generated 3HKA-like compounds, predicted to have an anti- inflammatory effect. These analogs will be screened in a series of in vitro and in vivo assay to fully assess their potential as novel immunomodulatory molecules

NIH Research Projects · FY 2025 · 2023-07

As an emergency/internal medicine physician who has served in several hurricane-impacted zones, my long-term goal is to become an independent physician-scientist whose research informs policies that mitigate the adverse health effects of extreme weather events (EWEs). The focus of this K08 is to receive mentored training to evaluate and improve programs like Department of Health and Human Services emPOWER program – the sole federal program designed to identify individuals at risk of premature morbidity and mortality due to EWEs. In 2021, EWEs adversely affected 1 in 3 Americans. The effects of hurricanes on cardiovascular health (heart failure, myocardial infarction, and stroke) are well-documented, and are likely to disproportionately affect such vulnerable populations as older adults, those with multiple comorbidities, and persons of low socioeconomic status. However, targeting such populations with U.S. federal pre-disaster mitigation is challenging because of difficulties identifying those at greatest disaster-related cardiovascular disease (CVD) risk and uncertainty about the timeframe for which risk is increased. We hypothesize that emPOWER can better identify those at risk by considering disaster-related CVD outcomes, incorporating available clinical, demographic, socioeconomic, and environmental factors into risk assessment, and considering these outcomes over longer time periods. This will enable emPOWER and local health system programs to more accurately identify those at high risk of adverse CVD outcomes, so that future programs can build tailored approaches to evacuate high risk groups before hurricanes strike and address the needs of those at excess CVD risk afterward. With mentored training in spatial analysis, CVD and social epidemiology, machine learning, and disaster science, I will evaluate the impact of Hurricane Sandy in 2012 on NYC because of its large population and neighborhood-level SES disparities-- and because no further hurricanes affected NYC during the study period, accomplishing the following aims using Medicare claims and ancillary datasets: 1) Estimate the hurricane’s impact up to 5 years after landfall on census tract-level CVD point prevalence in the ≥ 65 years old population using spatiotemporal modeling; 2) Compare the hurricane’s impact on CVD event rates for Medicare beneficiaries currently designated as high risk by emPOWER compared to all other beneficiaries using survival analyses and Cox models; and 3) Develop and internally validate a CVD risk prediction tool using machine learning techniques, and determine its implications. Guided by a board of federal and local disaster policy experts and mentored by nationally recognized experts, this K08 will provide me with the training and resources necessary to develop a CVD risk prediction tool to identify the most vulnerable individuals currently ignored by federal disaster planning policy, and allow me to build a career that improves and evaluates disaster policy and interventions across various extreme weather events using science-based approaches to prevent adverse CVD outcomes driven by changing weather patterns.

NIH Research Projects · FY 2024 · 2023-07

ABSTRACT Mycobacterium tuberculosis (Mtb) was the world’s single leading cause of death from infection before COVID- 19. Direct-acting antimycobacterial regimens are long, often toxic and plagued by emergence of drug resistance. Adjunctive therapies could potentially speed the cure of tuberculosis by targeting a process in the host that alters the host-pathogen interaction to the advantage of the host. One potential form of host-directed therapy would be to help macrophages better survive Mtb infection. In vitro, the death of Mtb-infected mouse macrophages is co- dependent on a type I interferon (IFN), IFN-beta, that macrophages produce in response to Mtb, along with an additional contribution by Mtb. This application seeks to identify specific molecular participants in Mtb-induced macrophage death that are downstream of the type I IFN receptor. We have identified potential molecular participants by two approaches—a CRISPR activation screen that restored Mtb-induced cell death to macrophages lacking the type I IFN receptor, and an innovative biochemical pulldown approach using a probe based on a small chemical compound that rescues Mtb-infected macrophages from Mtb-induced cell death without impairing the growth of Mtb. This application aims to validate the candidates genetically, test the ability of already existing inhibitors to recapitulate the effect of knocking them out, and then place the validated candidates on a mechanistic path. That would set the stage for future studies, beyond the scope of this application, to find, improve or develop drug-like inhibitors for tests in preclinical models of TB to see if they mitigate pathology and hasten cure when used in combination with direct-acting antimycobacterial agents.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY / ABSTRACT For patients with thoracic aortic aneurysms (TAA), replacement of the ascending aorta with a current standard of care prosthetic graft (polyethylene terephthalate) eliminates risk for dissection in graft-replaced regions and can thus be lifesaving. Nevertheless, accumulating evidence reveals that proximal aortic grafting can increase risk for downstream dissection, which is also life-threatening: Risk is greatest in patients with genetically triggered TAA, who undergo graft replacement at lower thresholds, higher frequency, and younger age - after which risk for dissection in graft-replaced regions is eliminated but possibility of distal complications increases. Up to two thirds of dissections in genetic TAA patients occur in the distal (arch or descending) aorta. We have also shown that over half of distal dissections with genetic TAA occur after graft surgery; proximal grafting has been linked to a >2-fold increase in risk for dissection independent of aortic size. Our clinical observations are consistent with experimental data: In pre-clinical and computational models, the dramatic increase in proximal aortic stiff- ness with grafting induces hemodynamic changes that exacerbate distal stiffening. Aortic stiffness is increased with genetic TAA - it is also known that mechanical loading forces drive adverse aortic remodeling. There is thus a critical need to identify markers of distal aortic disease progression after proximal grafting, with focus on altered hemodynamic loading in relation to graft characteristics (stiffness, length, enclosed volume). Our central hypoth- esis is that loss of proximal aortic compliance due to stiff prosthetic grafts induces adverse distal aortic remod- eling (driven by increased wall and wall shear stress) and predicts adverse prognosis. We also posit that adoption of grafts, for which compliance is tailored to compensate for patient-specific aortic stiffness, will attenuate ad- verse distal aortic remodeling. This will be tested in genetic TAA patients undergoing prosthetic graft replace- ment, via Aims integrated towards the goal of testing if graft implantation produces progressive increments in adverse remodeling (Aim 1A), identifying (native aortic and graft) features most responsible for adverse remod- eling (1B), testing if these features are modifiable via a new class of tailored grafts (Aim 2), and exploring if widely generalizable surrogates of graft-induced remodeling and native aortic stiffness predict clinical events (Aim 3). To do so, cardiac MRI will be integrated with computational modeling of fluid structure interactions and vascular remodeling - informed by material property testing of resected aortic tissue and simulations of tailored grafts for which compliance can be paired to patient-specific aortic features. Our team provides complementary expertise in cardiac imaging, aortic surgery, genetic TAA, computational modeling, and graft design - and a track record of productive collaboration. Results will yield key foundational insights as to mechanisms of adverse remodeling and events after grafting, transform risk stratification for current grafts and inform personalized therapy by guiding design and prototyping of a new class of tailored grafts - towards the goal of improved outcomes for TAA patients who benefit from proximal grafting but remain at risk for serious clinical events in the residual native aorta.

NIH Research Projects · FY 2025 · 2023-07

Summary Adult zebrafish have a remarkable capacity to regenerate the heart with minimal scarring. Understanding the underlying cellular and molecular mechanisms will help addressing the regenerative deficiency in the adult mammalian heart. We recently found that the zebrafish epicardium (the outermost layer of vertebrate hearts) regenerates after injury by the creation of a leader region of polyploid cells (having two or more copies of the genome). Polyploidy has been observed in many mammalian organs following injury and recently has been invoked in mechanisms of tissue repair. However, the functional significance of polyploidy, as well as its underlying mechanisms in tissue repair, remains elusive, representing a major knowledge gap in harnessing the advantages of polyploidy in tissue repair. We found that, through collective cell migration, these leader epicardial cells guide a trailing population of much smaller, dividing follower cells to repopulate the wound. The leader cell population is established and maintained by endoreplication and is eliminated through apoptosis upon completion of regeneration, indicating a transient role. The elevated cellular tension in the leader cells drives endoreplication. This coordinated behavior of leader and follower cells facilitates robust regeneration of the epicardium. Also, we found that the polyploid epicardial cells are a major source of paracrine secretion for heart regeneration. The overall objective of our proposal is to understand the mechanisms that regulate spatiotemporal cell behavior of the epicardium and how defects in this behavior impact heart regeneration. Through single-cell RNA sequencing, reporter assays, and pharmacological treatments, we have discovered a novel signaling pathway together with Yap signaling that participate in the spatiotemporal polyploidization in the epicardium. We will 1) characterize the signaling cascade that involves mechanical cues, Yap, and the new pathway in regulating spatiotemporal polyploidization during epicardial regeneration, 2) define the leader signals that drive leader- follower coordination in epicardial regeneration, and 3) investigate the functional significance of epicardial polyploidy in heart regeneration. The proposed research will define a new signaling paradigm in guiding cell cycle decisions for efficient heart regeneration. Moreover, polyploid cells are present in normal tissues such as the mammalian cardiomyocytes, as well as in pathological processes such as lung injury, acute kidney injury, and cancer. Results from our study will unearth conceptual innovations concerning the regulation of cell cycle decisions to mediate physiological and pathological polyploidization and robust tissue regeneration.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Monitoring the transition to wakefulness is critical during restoration to consciousness after brain injury, anesthesia, and in those COVID-19 survivors that have altered consciousness. However, we have an imprecise understanding of neural dynamics linked to behavioral changes as subjects awaken. Our previous work discovered that stimulating the anterior nucleus gigantocellularis (aNGC) promotes arousal from a coma-like state. We proposed recruiting multiple arousal pathways through aNGC as an avenue to triggering widespread activation resulting in wakefulness. Notably, aNGC activation increased frontal-motor cortical activity and restored full mobility through modulation of an aNGC-to-frontal-motor-cortex pathway despite high anesthetic concentration exposure. We also showed that animals emerging from diverse coma-like states share a common dynamic process of cortical and motor arousal that can be consistently sequenced from deep to high arousal levels. We identified five cortical periods that tracked restored motor behavior in a hypoglycemic coma and a range of anesthetics, whether inhaled or injected, alongside conventional righting reflex assays. Based on these findings, we postulate that restoring waking is a common progressive process in which cortical patterns contain metrics of consciousness that distinguish reflexive from purposeful movements. We hypothesize that cortical measurements that link neural responsiveness to defined behaviors are an applicable method that can extend the analysis of the recovery of consciousness beyond monitoring reflexive movements. Our proposal deepens our understanding of the contribution of cortical neural subtypes, the neuronal pathways underlying aNGC-induced changes in frontal-motor cortical activity, and the temporal dynamics that distinguish reflexive from the initiation of voluntary behaviors in our rodent-low arousal models. In addition, we will establish the cortical patterns that unpack these behavioral transitions. Since pathological states of unconsciousness are vastly heterogeneous, having a clear understanding of ordinary recovery serves to better appreciate the variability imposed by the injury to cortical activity and behavior. Thus, we will identify how damaged neural circuits affect established cortical activity pathways and dynamics that underlie behavior recovery. The proposed studies are thus significant because they will establish the mechanistic correspondence, examining activation of neural pathways and their dynamics linked to habitual and intentional behaviors that reveal novel, medically relevant biomarkers that promote a robust inference of arousal states during emergence from anesthesia and after brain injury.

NIH Research Projects · FY 2025 · 2023-07

One third of B-cell lymphoma patients relapse and remain incurable despite effective targeted therapies. Although, the serially relapsing nature of these tumors support the presence of stem- like lymphoma repopulating cells, this notion remains controversial and underexplored. Resistance to this concept arise from the fact that -in contrast to leukemia or other solid tumors that originate from stem-like cells- most lymphomas arise from fully differentiated, mature B cells. However, our preliminary studies provide strong evidence for the existence of rare subpopulations of B-cells undergoing antigen-activation (in response to pathogens) with stem-like molecular features and functional properties in a T-cell dependent manner. Moreover, we found that specific lymphoma-associated mutations further enhance the preexisting stemness program and potential. We, therefore, hypothesize that a subset of mature B cells is transiently endowed with stem-like epigenetic features, which are hijacked by specific lymphoma drivers, and constitute the molecular basis of their increased tumorigenic potential and tumor repopulating capacity. To address this hypothesis, we have built an interdisciplinary team of collaborators with expertise in stem cell reprogramming, epigenetics, immunobiology, single-cell technologies and lymphoma research. We have devised an innovative and bold approach employing multiple cutting-edge single-cell and chromatin technologies, as well as ex vivo and in vivo functional assays that will allow us to (i) determine the key regulators that promote or prevent increased GC B-cell plasticity in normal and cancerous contexts, (ii) decipher the signal and inter- cellular dependencies that enable emergence of a GC stem-like state and key vulnerabilities and (iii) dissect the synergies between common lymphoma drivers with GC stem-like properties, contributing to aggressive disease and relapse. The discovery of B-cell stem-like features and subpopulations will be paradigm-shifting and have a tremendous impact on the way we understand and treat lymphomas, opening new avenues for the development of superior diagnostic and therapeutic strategies.

NIH Research Projects · FY 2025 · 2023-07

Project Summary The opportunistic fungal pathogen Aspergillus fumigatus presents a major health concern in immunodeficient and critically ill patients, with invasive disease contributing to high mortality rates. Infection with A. fumigatus elicits a diverse adaptive CD4 T cell response. Our preliminary data have demonstrated distinct TH1 and TH17 spatial neighborhoods within mediastinal lymph nodes (LNs) during A. fumigatus airway infection. This spatial organization positions activated CD4 T cells to receive tailored signals (e.g., antigens, cytokines) for optimal effector cell differentiation and function. Lymph node stromal cells (LNSCs) are known to provide guidance cues for immune cell trafficking during monotypic effector T cell responses. However, the mechanisms by which distinct TH1 and TH17 microenvironments are concurrently established within LNs remain unknown. Defining these mechanisms will offer insights into the functional importance of LN microenvironments in establishing human immunity to infection and inflammatory disorders for the identification of novel therapeutic strategies. Based upon our single-cell RNA-sequencing and flow cytometric analyses of FRC and effector T cell subsets during A. fumigatus challenge, we propose that (i) FRC subsets establish spatially distinct TH1 and TH17 neighborhoods by dynamically regulating chemotactic receptor-ligand axes, such as CXCR3:CXCL9/CXCL10 and CCR6:CCL20 and that ii) these neighborhoods can persist after pathogen clearance. We will address these hypotheses in a clinically relevant murine model of invasive pulmonary aspergillosis. In Specific Aim 1, we will characterize distinct spatiotemporal microenvironments formed by CD4 T cells and FRCs in A. fumigatus infection by applying spatial transcriptomic profiling followed by high-content immunofluorescence methods paired with a novel computational approach for image analysis. In Specific Aim 2, we will define the functional role of CD4 T cell spatial organization in A. fumigatus infection by perturbing FRC- dependent chemokine gradients in a cell type-dependent manner using FRC-specific gene targeting. We will also conduct adoptive co-transfers of A. fumigatus-specific CD4 T cells sufficient and deficient in CXCR3 and CCR6. In Specific Aim 3, we will investigate the durability of infection-driven spatial FRC diversity by studying epigenetic modifications in A. fumigatus-experienced FRCs. We will also employ high-content confocal imaging to determine whether prior infections affect FRC formation of T cell neighborhoods in subsequent infections. This study employs and develops novel genetic tools, microscopy methods, and computational approaches to generate a systems level understanding of secondary lymphoid organ immunobiology. Furthermore, this proposal is tailored for a physician-scientist in training as it investigates the mechanisms by which stromal cells induce adaptive immunity to the clinically relevant pathogen A. fumigatus, with implications for anti-fungal therapeutic strategies and vaccine development.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Nemaline myopathy (NM) is a skeletal muscle disease hallmarked by muscle weakness with an incidence of 1 in 50,000 live births. On histopathology, there is an obvious presence of actin accumulations in disrupted muscle. The causative mutations identified thus far are in genes critical for actin filament structure within the muscle, yet the molecular mechanisms for how alteration of these proteins leads to NM pathology is not well understood. Cofilin-2, which is important for actin filament severing, is one such affected actin-binding protein. This cofilin isoform is the predominant form in postnatal and mature skeletal muscle; its function has mainly been studied with respect to actin at the sarcomere, the muscle’s main contractile unit. Cofilin-2 is known to biochemically bind more readily to cytoplasmic non-sarcomeric actin than sarcomeric actin, but the impact of this characteristic on NM progression has not been studied. A Drosophila model of muscle-specific cofilin (DmCFL) knockdown was shown by our lab to have progressive muscular defects linked to sarcomere addition during growth. I analyzed RNA sequencing data produced from muscle-enriched preparation of the DmCFL knockdown model and found that genes associated with excitation-contraction coupling (ECC) are differentially expressed. ECC is the process by which signals from the motor neuron are communicated to the muscle ultimately leading to contraction. My preliminary data show disordered actin organization at the muscle side of the neuromuscular junction (NMJ), which is where the muscle receives signals from the motor neuron. Based on the literature and these preliminary findings, I hypothesize that cofilin regulation of non-sarcomeric actin is critical for the proper NMJ and contraction machinery structure needed for ECC prior to muscle deterioration. To address this hypothesis, I will use the DmCFL knockdown model to analyze the impact of decreased cofilin on the NMJ signal transduction (Aim 1) and muscle contraction (Aim 2). The former will be accomplished by using molecular, microscopy, and electrophysiological techniques to analyze changes in NMJ protein localization, morphology, and function at the muscle (Aim 1A) and motor neuron (Aim 1B). I will compare the morphological findings from Drosophila larval muscle to those from cofilin-2 knockout mouse muscle samples. Contraction will be assessed using fluorescent imaging techniques targeted to the calcium signaling machinery (Aim 2A). Using a modified exercise approach (Aim 2B), I will discover how exercise intensity influences contraction and phenotype progression in DmCFL knockdown larva. These experiments will collectively provide insight into the status of the NMJ and contractile activity in cofilin NM while leveraging the simplicity yet high level of evolutionary conservation of Drosophila. In answering this question relevant to a clinical disease that typically manifests early in life, I will further develop the technical skills and scientific reasoning needed as a cell and developmental biologist. These studies will complement my clinical activities as I train to become a well-rounded physician-scientist and independent investigator interested in pediatrics.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY SARS-CoV-2 infections in children are mostly milder and less lethal than in adults. This lower incidence of relevant disease associated with SARS-CoV-2 has surprisingly also been observed for children with asthma, the most common chronic respiratory and inflammatory disease in children. As social distancing and masks have dramatically but temporarily decreased the spread of common viral respiratory infections, this created the positive effect of a significant decrease of viral-triggered asthma in children. The growing prevalence of SARS-CoV-2 infection in children and the resurgence of non-SARS-CoV-2 respiratory viral infections make it critical and timely to understand how SARS-CoV-2 infection shapes respiratory outcomes and immune responses to other respiratory viruses and the upcoming SARS-CoV-2 vaccines. In addition to a strong genetic predisposition, asthma is strongly linked to viral respiratory infections, and infectious stimuli can have long-term epigenetic consequences that shape immune responses to subsequent infections. We propose that a common genetic variation that alters sphingolipid levels in children with asthma may also result in limited pathogenesis of SARS-CoV-2 in children with asthma. This proposal aims to test the hypothesis is that common genetic variation in asthma moderate age-dependent outcomes and immune responses to SARS-CoV-2 infection and vaccines utilizing an NYC pediatric asthma cohort that was started early in the pandemic. This cohort is uniquely suited to the hypothesis as it (1) includes children of all ages with and without asthma; (2) is enriched for children from ethnic, racial, and socioeconomic backgrounds associated with health disparities and high exposure to SARS- CoV-2; and (3) has already enrolled more than three hundred with a high antibody positivity rate since May 2020. The availability of detailed questionnaire data on asthma and SARS-CoV-2 exposure, biospecimen that include blood (including stored PBMC), and nasal samples will enable us to address these three Specific Aims, to (1) determine the age-dependent effect of SARS-CoV-2 infection on subsequent respiratory health in asthmatic children; (2) define the epigenetic and transcriptomic effect of SARS-CoV-2 infection on hematopoietic stem cells, and (3) define the effects of common genetic asthma risk alleles on responses to SARS-CoV-2 infection of nasal epithelial cells and subsequent infection with respiratory syncytial virus and rhinovirus. These studies will be supported by a team with expertise in viral immunology, pediatric asthma, and epigenetics of immune responses and will inform on age- and disease- specific impact and mechanisms of SARS-CoV-2 and common respiratory viruses in children.

NIH Research Projects · FY 2025 · 2023-07

Project Summary/Abstract In 2020, with over 250 million debilitating cases and over half a million deaths, mostly in young children, malaria is a persistent global health crisis. The malaria-causing parasite Plasmodium falciparum (Pf) has developed resistance to most antimalarial drug deployed, including the backbone artemisinins (ARTs). ART and its semi-synthetic analogs are considered essential for malaria treatment. ARTs are prodrugs that are activated within the parasites to form a reactive radical that covalently attacks proteins, lipids and other cellular constituents. ART resistance is widespread in Southeast Asia and has been reported in Africa. ART combination therapy (ACT) is a mainstay for treatment of malaria, but its efficacy can be derailed when a two-drug combination becomes de facto monotherapy. Moreover, extended exposure of Pf to ACTs induces multidrug tolerance. We recently showed that inhibitors specific for the Pf proteasome (Pf20S) kill Pf in each stage of its life cycle and synergize with ART, overcoming ART resistance. This proposal builds on our discovery that a covalent hybrid of an ART analogue and a Pf20S inhibitor that we call an artezomib (ATZ) can enhance ART action and overcome resistance to each of its components. We have synthesized ATZs that are more potent Pf20S inhibitors than their component Pf20S inhibitor. They not only kill wild type and ART-resistant (K13 mutant) Pf, Pf with proteasome mutations that confer resistance to the Pf20S inhibitor, but also kill Pf that expresses both ART-resistant and PI-resistant mutations. We propose the following mechanism by which ATZs overcome resistance to the Pf20S inhibitor within them: We found that upon activation of ATZ in the parasites, the ART component binds Pf proteins, like activated ART itself. The Pf ubiquitin proteasome system digests ATZ-bound proteins into oligopeptides, some of which display the Pf inhibitor component of the ATZ. We hypothesize that extended contact of ATZ-bearing peptides within the Pf20S active site augments the binding of the Pf20S inhibitor component of the ATZ, overcoming the decreased binding otherwise conferred by Pf20S point mutations. Thus, an ATZ can overcome resistance to each of its components. In mouse models of malaria, an ATZ drove P. berghei below the limit of detection and suppressed recrudescence of a P. berghei ART-resistant K13 mutant and doing so better than ART. In Aim 1 of this proposal, we will conduct lead optimization to improve ATZs' potency, selectivity and ATZs' pharmacokinetic properties. In Aim 2, we will explore ATZs' mechanism of action; attempt to select for ATZ-resistant parasites; determine the frequency and mechanism of resistance, if any; and study antimalarial activity of ATZs in stages of the Pf life cycle when ART alone is ineffective. Aim 3 will test the efficacy of ATZs in mice, including humanized mice infected with Pf. .

NIH Research Projects · FY 2025 · 2023-07

Project Summary – Immunometabolic Programs Controlled by ER Stress in Cancer Tumors create hostile microenvironments that impede the development and maintenance of effective anti-cancer immunity. Yet, how intratumoral immune cells integrate and interpret persistent stress signals in this harsh milieu remains incompletely defined. We have uncovered that adverse conditions within malignant masses disrupt the protein-folding capacity of the endoplasmic reticulum (ER) in infiltrating immune cells, triggering dysregulated “ER stress” responses that promote immunosuppression and malignant progression. Therefore, we postulate that understanding and targeting detrimental ER stress responses in the tumor microenvironment represents a major opportunity to develop new and more effective forms of cancer immunotherapy. In a key recent advance, we determined that the IRE1a-XBP1s arm of the ER stress response inhibits the expression of Tagln2, an understudied cytoskeletal protein implicated in T cell activation and effector function. We found that XBP1s-deficient T cells demonstrate enhanced Tagln2 expression that supports their protective function at tumor sites. Moreover, we established that Tagln2 coordinates major metabolic programs that sustain robust T cell mitochondrial respiration and effector capacity. These new findings have prompted us to dissect the mechanisms by which the novel IRE1a-XBP1s-Tagln2 axis controls T cell metabolism and function in cancer. Hence, the goals of this project are to i) understand how ER stress inhibits Tagln2 expression, ii) establish that Tagln2 equips T cells with robust metabolic fitness that supports their anti-tumor activity, and iii) determine that Tagln2 replacement therapy enhances the activity of chimeric antigen receptor (CAR)-T cells in solid tumors. We hypothesize that dysregulated ER stress responses hinder intratumoral T cell function by disabling Tagln2- driven bioenergetic programs, and that sustaining this cytoskeletal-mitochondrial axis could be used to improve the efficacy of adoptive T cell immunotherapy in solid tumors. We will test this novel hypothesis in the setting of immunotherapy-refractory ovarian cancer (OvCa) through the following Specific Aims: Aim 1. Define the mechanisms by which ER stress responses inhibit Tagln2 in OvCa-infiltrating T cells. Aim 2. Establish the role of Tagln2 as a metabolic driver of competent anti-tumor T cell function. Aim 3. Test the hypothesis that preserving Tagln2 activity enhances CAR-T cell immunotherapy in OvCa. Collectively, the proposed project will expand our mechanistic understanding of immune regulation in the tumor microenvironment and promises to pave the way for novel interventions that augment the efficacy of cellular immunotherapy against solid malignancies.

NIH Research Projects · FY 2025 · 2023-06

Abstract: Despite the irreplaceable contributions of physician-scientists to clinical, the pipeline of clinician-scientist trainees remains on the decline. A critical barrier to the physician-scientist career is the lack of protected time and access to research mentorship available to physicians during clinical training. With the heightened acuity, volume and complexities of modern medicine, clinical training has increasingly focused on the acquisition and application of existing, rather than generation of new, knowledge. An unfortunate consequence of this shift in focus is the increasing compartmentalization of scientific training into fragments of time that are inadequate to provide the training and continuity needed to become proficient in the scientific investigation. The Tri-Institutional Stimulating Access to Research during Residency (Tri-I StARR) program seeks to remedy this deficiency by developing an integrated, longitudinal mentored research training program that will lead to the development, implementation, and evaluation of new clinical interventions to prevent, diagnose, treat and ameliorate health disparities of disorders of infectious, immunologic and inflammatory etiologies, with mentorship from across Weill Cornell Medicine, Rockefeller University, and Memorial Sloan Kettering. Tri-I-StARR will train residents on an integrated clinical-research pathway across 3 departments: Pediatrics, Medicine, and Pathology in areas along the full biomedical research continuum and include the themes of healthcare disparities and health and disease over the life course. The program proposes four training aims: (1) Acquisition of skills in the scientific method and design of hypothesis-based projects to address human diseases across the lifespan and their inequities, (2) Individualized, multidisciplinary mentorship in the design and completion of a research project, (3) Development of short, intermediate, and long term IDPs that integrate scientific and clinical training within and across career stages, and (4) Active engagement in horizontal and vertical networking among physician-scientists within and across career stages and institutions. The Tri-I-StARR will be led by an Executive Committee (EC) consisting of Kyu Rhee, MD PhD (Medicine), Sallie Permar, MD, PhD (Pediatrics), and Ethel Cesarman, MD (Pathology), an Expanded Executive Committee (EEC) of Residency Program Directors and Program Coordinators, and a team of 36 multi-departmental, multi-disciplinary, well-funded, and experienced faculty preceptors. Three Resident-Investigators each year will be supported for 12 months of research with the options to add an additional 12 months. Upon completion, trainees will be capable of transitioning to research-intense fellowship training, successfully competing for follow-on funding opportunities, including the K38, that will enable them to become the next generation of physicians leading and mentoring trainees in clinically-oriented research. This program will fulfill the urgent need for: 1) more full-time academic physician-researchers in medical schools throughout the country, and 2) innovations and clinical translation of novel strategies to improve health across the lifespan.

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY The goal of this proposal is to discover how ongoing chromosome missegregation events in cancer cells (a process called chromosomal instability, or CIN) alters the tumor ecosystem to promote cancer progression. Chromosome copy number alterations (also referred to as aneuploidy) have long been associated with immune suppressive phenotypes, drug resistance, and metastasis. Beyond aneuploidy, however, it remains unknown whether the ongoing process of chromosome missegregation gives rise to tumor progression. Harnessing ex- perimental tools that enable us to dial-up or dial-down chromosome missegregation rates in otherwise isogenic backgrounds, we have previously found that ongoing chromosome missegregation generates rupture-prone mi- cronuclei, which expose genomic double-stranded DNA (dsDNA) to the cytosol, leading to persistent activation of the cGAS-STING innate immune pathway (1). Yet, instead of promoting a robust type I interferon (IFN) re- sponse, STING activation in cancer cells with CIN promotes noncanonical NF-kB (nc-NF-kB) signaling – through an unknown mechanism – to drive metastasis. The extent to which CIN-driven metastasis is dependent on the immune system remains unknown. We made the surprising observation that CIN drives tumor progression in a cancer cell non-autonomous manner by shaping the interaction between cancer cells and the tumor microenvi- ronment (TME). This finding motivated the development of a fundamentally new, systems-level approach to evaluate the nature and conditional-dependence of cell-cell interactions in the TME called ContactTracing. This method exploits intrinsic biological variance captured by single cell RNA sequencing technologies, to infer cellu- lar responses to ligand-receptor mediated interactions without prior knowledge of downstream target genes. Combining this innovative computational tool with genetic perturbation of CIN and STING we found that CIN engenders a pro-metastatic TME by inducing a cancer cell-intrinsic ER-stress response. In Aim 1, we propose to mechanistically dissect the epistatic relationship between CIN, STING, and ER-stress in the progression of triple negative breast cancer (TNBC) to determine whether ER-stress can represent a therapeutic target in chro- mosomally unstable tumors. We will also test whether an ER-stress response underlies nc-NF-kB activation. Under Aim 2, we will improve causal inference of tumor-derived ligand effects and explore their molecular basis using gene regulatory networks to ask whether CIN-dependent cell-cell interaction networks are conserved across cancer types using both human data and mouse models of breast, pancreatic, and lung cancers. The amalgamation of these approaches, combined with our deep understanding of CIN in cancer, is poised to eluci- date the complex roles of CIN-induced STING signaling on tumor-immune crosstalk during disease progression. Importantly, this work is poised to reveal novel strategies aimed at targeting chromosomally unstable tumors, which are otherwise difficult to treat.

NIH Research Projects · FY 2026 · 2023-06

The development of TB drugs benefits greatly from validation of novel drug targets in predictive animal models. M. tuberculosis (Mtb) enzymes in central carbon metabolism are emerging as promising targets for drug development but have not been validated in animal models that recapitulate the diverse and heterogeneous environments Mtb encounters in humans. We propose to test the hypothesis that these heterogeneous environments result in airway, lung and granuloma-dependent nutritional restrictions that, at times, make Mtb dependent on both glycolysis and gluconeogenesis to establish and maintain infection in nonhuman primates and/or during paucibacillary infection in mice. We will infect NHPs with Mtb mutants of two enzymes – PFK and PEPCK - that are required for glycolysis or gluconeogenesis, respectively. We will use deletion mutants to determine whether these enzymes are required for establishment of infection and use conditional knockdown mutants to investigate the enzymes importance for growth and survival in different pathologies and for progression of disease. We will furthermore evaluate these mutants in a relapse mouse model to test the hypothesis that the nutritional requirements during long-term persistence in mice, when the bacteria cannot be cultured in vitro, are different from those encountered during active growth and high titer chronic infection and ask whether they mimic environments encountered in NHPs. This proposal builds on the experience of the multi- PI team in mycobacterial genetics, metabolism, and animal models, with the goal to dissect the carbon source requirements for Mtb in the model system that is most similar to human Mtb infection and disease.