Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2024 · 2023-04

PROJECT SUMMARY Chronic kidney disease (CKD) affects 10% of the population worldwide. More than 37 million people are estimated to have CKD in the US, and 2 in every 1000 Americans need dialysis or a kidney transplant to survive. CKD has numerous systemic complications including anemia and dysfunctional systemic iron homeostasis. Kidney fibrosis is the final mechanism common for all progressive kidney disorders. Unfortunately, very few therapies are available to slow the progression of kidney fibrosis in patients with CKD. Kidney macrophages are one of the key cells implicated in the pathophysiology of kidney fibrosis. The majority of kidney macrophages originate from circulating monocytes. In his K08-funded project Dr. Akchurin elucidated the pathologic role of depletion of labile iron pool in kidney macrophages in propagation of kidney fibrosis. This R03 proposal will enhance his capabilities of deciphering the role of iron in pathologic behavior of myeloid cells in CKD by now, in addition to kidney macrophages focusing on their precursors, circulating monocytes. Based on the preliminary data, the central hypothesis of this proposal is that in patients with CKD, circulating monocytes, similar to kidney macrophages, exhibit pathologically depleted intracellular labile iron pool LIP, which alters their phenotype by inducing CX3CR1 and other receptors facilitating chemotaxis and tissue invasion. Repletion of this intracellular monocyte labile iron pool through correction of systemic iron imbalance will be associated with reduction of this pro-inflammatory differentiation. This hypothesis will be tested in two specific aims: (1) determine the intracellular iron status of classical, intermediate, and non-classical circulating monocytes in patients with CKD and (2) elucidate the role of systemic and intracellular iron status in the differentiation of monocytes in patients with CKD. To test this hypothesis, the applicant will use the existing repository of peripheral blood mononuclear cells collected from children with CKD with and without functional or absolute iron deficiency and from healthy control children. He will evaluate these cells using single cell transcriptomic and multicolor flow cytometry approaches with the focus on delineating the phenotypic features responsible for subsequent tissue invasion, such as expression relevant chemokine receptors, including CX3CR1. Furthermore, he will perform ex vivo evaluation of human monocytes collected from CKD patients to directly assess their relevant functional characteristics in the presence and absence of iron stimulation. Monocyte functional characteristics will be analyzed I the context of clinical parameters, levels of circulating and urine-excreted chemokines, and systemic parameters of iron homeostasis. Results techniques an kidney from t he outlined experiments will provide preliminary data, as well as experience in for the functional ex-vivo testing of uman monocytes, to support the K08 awardee's application for R01 proposal to evaluate the impact of iron metabolism and iron therapy on myeloid cells in the context of fibrosis and CKD progression. h

NIH Research Projects · FY 2026 · 2023-04

Summary: At least 400 older adults a day are discharged from US emergency departments (EDs) and within 72 hours experience a return ED visit resulting in hospital admission (RVA). Geriatric RVA have dramatically higher morbidity and mortality than patients admitted to hospital on their initial ED visit. These outcomes, combined with the clinical complexity of geriatric presentations, demonstrate a critical need for clinical decision support (CDS) for ED discharge decisions and improved post-ED care management in older adults. National guidelines recommend that all older adults receive formal risk screening in the ED. Existing geriatric ED risk assessment tools lack predictive validity and are not designed to identify the multifactorial risk of an RVA event within 72 hours after ED discharge. Our long-term goal is to improve the outcomes of older adults using machine learning models for clinical decision support (CDS) in emergency medicine. The goal of this study is to develop and validate a machine learning model that predicts geriatric emergency medicine 72-hour RVA (GEMRA), and can be used as a feasible ED CDS tool. In order to maximize the impact and generalizability of GEMRA across a wide range of US ED environments and populations, the model input variables used will be clinical data collected in the course of normal clinical care, and thus widely available in emergency health records (EHRs). GEMRA will be developed and validated with data from five diverse hospitals across two health systems that span a wide range of demographic, socioeconomic, and ethnic backgrounds. The study will be conducted by a closely collaborating interdisciplinary team that includes emergency medicine, machine learning, and CDS experts, with extensive experience in geriatric emergency medicine research as well as developing and evaluating technological driven interventions to improve post-ED outcomes. Our preliminary work demonstrates that an early machine learning model using 478 clinical data input variables can accurately identify ED patients at high risk of RVA, outperforming an existing, unvalidated traditional RVA risk score that used six clinically derived risk factors. Our specific aims include: (1) Optimize GEMRA through model refinement, validation with retrospective data from unseen populations, as well as explanation of model performance variation across different clinical subgroups; (2) Assess GEMRA's clinical value through prospective validation at three different hospitals, comparing model performance to existing ED geriatric and RVA risk tools, as well as real-time clinician judgment; (3) Engage multidisciplinary stakeholders in the design of both a GEMRA CDS prototype and a complementary multidisciplinary clinical RVA risk assessment workflow; and subsequently evaluate the feasibility of these products in ED clinical practice during a short-term pilot implementation study. Completion of these aims could transform older adult post-ED risk screening, leveraging the computational power and scalability of machine learning to identify patients at risk of early post-ED adverse outcomes. Subsequent implementation of GEMRA CDS would inform risk-mitigating interventions, potentially impacting outcomes in this vulnerable population.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY / ABSTRACT On average, a man dies from PCa every 16 minutes, mainly due to development of secondary malignant growths outside of the primary cancer site, known as metastases. The cornerstone of PCa treatment is androgen deprivation therapy (ADT). ADT temporarily halts PCa, but leads to resistance in nearly all cases, resulting in castration-resistant PC (CRPC). CRPC then undergoes further evolution of metastatic subclones and results in incurable disease. Research techniques revealing resistance mechanisms and clonal evolution of metastatic PCa are lacking due to the limited capacity of current animal models to mimic PCa evolution in its native microenvironment as well as inefficient methods for tracing subclonal evolution. Therefore, we developed EvoCaP (!Evolution in Cancer of the Prostate”), a mouse model of endogenous metastasis that recapitulates human PCa genetically, by using PTEN/TP53 co-deletions enriched in metastatic patients, and phenotypically, by focal initiation of primary disease progressing to bones, lungs, lymph nodes and liver metastases. Our model uses a lentiviral platform - LV.CreBC10 carrying: (1) Cre (Pten/Trp53 co- deletions; activation of Cas9, fluorescence and luminescence markers); (2) Barcode with ten sites for marking by Cas9 (BC10); (3) RNA guide specifically marking BC10; and (4) guide or short hairpin RNA for testing metastatic drivers. Luminescence (FLuc) permits continuous tracking of disease progression and fluorescence (eGFP) allows for specific sorting of cancer cells. BC10 represents a synthetic array of on-target sites, in order of decreasing activity, for the RNA guide that attracts Cas9 to generate subsequently specific edits. To streamline barcode analysis, we have established an R package - EvoTraceR. This comprehensive system enables: (1) the profiling of cancer cells based on shared mutational patterns in primary and metastasis; and (2) the building of phylogenetic trees to track evolution toward metastases in a robust and flexible way. Our central hypothesis is that differences in distinct molecular and phenotypical clonal architectures will be precisely detected between primary and metastatic sites depending on therapy status, enabling the inhibition of metastasis and/or resistance promoting genes and pathways. Our analyses will establish and mechanistically validate drivers of metastatic clonal expansion caused by Pten/Tp53-loss (basal) and also investigate how evolutionary pressure from therapy (ADT), applied at different stages of PCa, leads to the emergence of resistant clones. We will then use Cas9/guide (g)RNA and inducible short hairpins to target genes altered in those expanding clones to identify drivers of both treatment-naive and treatment-induced PCa metastasis. EvoCaP can feasibly track molecular evolution and validate targets for drug development, which may lead to identification of novel metastatic driver genes and pathways. Thus, therapies could be applied in: (1) primary diseases for early detection and interruption of metastases development; and (2) already existing metastases. Importantly, technologies developed in this project can also be applied to other types of metastatic cancers.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY This proposed postdoctoral training program (2 fellows/year, 2-year program) in endocrinology and metabolism will provide comprehensive research training for individuals with a serious commitment to a career at the interface of biomedical research and endocrine disorders. Metabolic diseases represent significant health burden in the US; advances in the pathophysiology of these diseases are required to improve diagnostics, therapeutics and health outcomes. Weill Cornell Medicine (WCM) is experiencing accelerating growth in clinical and biomedical research activities, due to the singular emphasis on developing medical research over the past decade during the Deanships of Dr. Laurie Glimcher and Dr. Augustine Choi. Investment in the Department of Medicine and the Division of Endocrinology, Diabetes and Metabolism has been significant. Dr. Anthony Hollenberg, a prominent endocrinologist and thyroid biologist, was recruited to Chair the Department of Medicine in 2018; his support has catapulted metabolic research to a high priority on campus, resulting in recruitments and significant internal and external resources. Dr. Laura Alonso, islet biologist and PD on this proposal, was recruited in 2019 to be Chief of the Endocrinology Division and Director of the Weill Center for Metabolic Health, a newly assembled research group on campus focused on metabolism, with 45+ faculty and 150+ members. Our proposed T32 leadership team also includes Dr. Julianne Imperato-McGinley, renowned endocrinologist, prior Division Chief and current Director of the Weill Cornell CTSC. However, the impetus for this proposal is not the leadership team, but rather the incredible trainees that have been finding a path to academic medicine even without T32 support. Three clinical endocrine fellows in the past 10 years managed to obtain K08 awards and are currently leading successful academic research careers, despite the lack of an available T32; one of our current 2nd year fellows elected to pursue a 3rd year of research training even though our program has been advertised as a 2-year clinical program, and both of the incoming clinical fellows for 2022 are outstanding researchers, one with a PhD from Cal Tech and the other with a first author Nature Medicine paper in islet biology, mentored by one of our faculty preceptors (Dr. Lo). We have hand-picked a top team of research Preceptors, and designed a comprehensive didactic training plan with wrap-around mentoring to guide trainees through eventual R01 submission. We propose a tri-institutional (WCM, MSKCC and RU), rigorous multidisciplinary T32 to enroll 2 postdoctoral MD or PhD trainees annually for 2 years training duration each. The leaders of this training program will work with the utmost enthusiasm and energy to ensure that our trainees reach their potential to excel and contribute significantly to the academic endocrine community for years to come.

NIH Research Projects · FY 2025 · 2023-04

Project Summary Type 2 diabetes (T2D) is caused by impaired β-cell insulin secretion and reduced β-cell mass. Both features have been linked to the failure to maintain β-cell identity. As a result, functional β-cell cells dedifferentiate into non-functional endocrine progenitor-like cells. In this regard, whether β-cell dedifferentiation is reversible is one of the most important notions in terms of disease modification. Aldehyde dehydrogenase 1 isoform A3 (ALHD1A3) has been discovered as a marker of β-cell dedifferentiation in diabetic mice and human T2D pancreata. ALDH1A3-positive β (A+) cells have been shown to be functionally defective. Moreover, pair-fed db/db mice showed improved glucose control associated with a significant decrease in the number of A+ b cells. The strong correlation between ALDH1A3 expression and β-cell function suggests that ALDH1A3 is more than a marker and may also plays a role in β-cell dedifferentiation during diabetes progression. Furthermore, it is unknown whether the decrease in A+ cell number under pair-feeding was due to reversal to a normal b-cell phenotype, death, or the emergence of new β cells. Answering these questions will provide clues on whether and how β-cell failure can be reversed. Thus, the PI first established two animal models to address these queries: one to investigate the role of ALDH1A3 in dedifferentiating β cells (β-cell specific Aldh1a3 knockout) and another to investigate the fate of ALDH1A3-expressing (A+) cells during pair-feeding (ALDH1A3 Creert knock-in lineage- tracing). The latter model will allow her to address whether A+ cells are converted back into ALDH1A3-negative (A-) cells with restored β-cell function. To complement these models, the PI will also use selective chemical ALDH1A3 inhibitors to see if ALDH1A3 blockade can reverse β-cell failure. In this application, she will test the potential therapeutic effects of genetic and pharmacological ALDH1A3 inhibition in diabetes in vivo and its mechanism of action (Aim 1). Using an ALDH1A3 lineage tracing mouse model, she will directly test the reversibility of β-cell dedifferentiation in diabetic mice in pair-fed condition, and in the treatment with ALDH1A3 inhibitors, KOTX1 and GA11 (Aim 2). The successful completion of this application will demonstrate the role of ALDH1A3 in β-cell function in the pathophysiological process of T2D. More importantly, the proposed work will assess whether ALDH1A3 is a potential therapeutic target in the treatment of T2D by reversing β-cell dedifferentiation/failure.

NIH Research Projects · FY 2025 · 2023-04

Resident-to-resident aggression (RRA) in long-term care (LTC) is associated with preventable injury, suffering, and serious psychological distress. One in five residents in nursing homes (NH) experiences RRA in a given month. Residents with Alzheimer’s Disease and Related Dementias (ADRD) are at an even higher risk of RRA due to cognitive impairment-related symptoms. However, RRA remains poorly understood. Differences in the backgrounds and life experiences of residents may play an important role in RRA initiation and escalation, risk/resilience factors, consequences, and NH response systems. The goal of this project is to systematically investigate differences in resident background and life experiences in the types, patterns, and circumstances surrounding RRA using the first prevalence cohort study of RRA, and collecting additional stakeholder input with explicit consideration of the needs of residents with ADRD, to develop, refine, and pilot-test a novel intervention. In Aim 1, I will leverage the first and only NIH-funded RRA prevalence cohort study to qualitatively contextualize and quantitatively examine the role of background and life experience differences between residents in RRA across individual, dyadic, and facility levels. In Aim 2, I will gather input from multiple NH stakeholders to improve understanding of the role of background and life experience differences in RRA, and to identify current and optimal intervention and prevention strategies in residents with and without ADRD via mixed methods approach. In Aims 3a and 3b, findings from Aim 1 and Aim 2 will be used to develop and refine a staff education intervention focused on RRA between residents from different backgrounds and life experiences that may be integrated into an existing RRA intervention program. As a social scientist trained in public health research, I am ideally positioned to spearhead this line of research, given my productive track record in elder mistreatment research. Through the award period, I will build upon my prior training to develop new knowledge and skills in long-term care research (Aim 1; Training Objective 1); research on population-level health differences (Aim 1; Training Objective 2); long-term care policy-making, advocacy, and leadership (Aim 2; Training Objective 3); and behavioral intervention and implementation science (Aim 3a, 3b; Training Objective 4). Career development activities will consist of formal coursework, experiential learning and research opportunities, and mentorship from experts in elder mistreatment, RRA research, and behavioral intervention development. This award will help me achieve my long-term career goal of being an independent investigator with an impactful program of long-term care research focused on impact-driven, evidence-based elder abuse prevention and intervention development and implementation for older persons with ADRD.

NIH Research Projects · FY 2026 · 2023-04

The aging brain is characterized by a slow deterioration of homeostatic balance between pro- and anti-inflam- matory cytokines, resulting in a proinflammatory state. Resident CNS immune cells are normally present in a resting state, but exhibit heightened vulnerability to secondary insults with aging, leading to a phenotypic shift in cell surface marker expression and cytokine release that contribute to the proinflammatory state. The functional contributions of interactions between cell surface proteins of neurons and immune cells have not been fully addressed. Using an unbiased protein-protein interaction screen, we determined that the immunomodulatory ligand B7-1 (CD80) interacts with the p75 neurotrophin receptor (p75). Importantly, the B7-1:p75 interaction is of recent evolutionary origin, present only in primates, although exogenously applied human B7-1 (hB7-1) binds murine p75 due to the extensive conservation of p75. We mapped the B7-1 surface responsible for p75 engagement and demonstrated that it includes regions known to interact with CTLA-4/CD28 and extends to additional surface regions. Given this overlap, CLTA-4/CD28 directly compete with p75 for binding to hB7-1. Exposure of murine hippocampal neurons in vitro to hB7-1 acutely alters dendritic and spine morphology, with loss of postsynaptic protein PSD95 and microtubule discontinuity in a p75-dependent manner. Abatacept, an FDA-approved therapeutic protein (CTLA-4-Fc), inhibits these effects. In vivo injection of hB7-1 into the murine subiculum, a hippocampal region affected in Alzheimer’s Disease, results in acute p75-dependent pruning of dendritic spines. To study the effects of hB7-1 in the intact animal, and in models of neurodegeneration, we developed a chimeric humanized B7-1 knock-in mouse. Our long-term goal is to develop monoclonal antibodies that specifically block hB7-1:p75 engagement for the prevention of neurodegeneration, and we have identified two such monoclonal antibodies that block hB7-1:p75 synapse elimination at 10nM concentration. Aim 1 will further evaluate and prioritize these and other monoclonal antibodies to develop additional reagents with broad epitope coverage. In Aim 2, our two lead antibodies, and other prioritized antibodies, will be evaluated using hippocampal neuron cultures for effects on blocking synaptic elimination and negative dendritic remodeling. Aim 3 will evaluate the in vivo effectiveness of select antibodies on preventing hB7-1 spine elimination and behavioral impairment by blocking the interaction of immune cell-expressed B7-1 with neuronal p75 in WT, chimeric B7-1 and an AD mouse model. These studies represent conceptual, mechanistic and therapeutic advances by (1) extending our understanding of immune:neuronal interactions that promote neurodegeneration; (2) identifying mechanisms that exist in humans, but not mice, to overcome existing limitations of murine models; (3) identifying preclinical therapeutics for aging populations.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract Chronic Obstructive Pulmonary Disease (COPD) is a destructive inflammatory lung disease that is the 4th leading cause of death in the U.S., killing more than 150,000 people yearly. Current therapies can improve symptoms and reduce hospitalization, but none modify this disease or stop its relentless progression. COPD is commonly caused by cigarette smoke (CS), but we do not yet fully understand the early events that trigger inflammation and tissue destruction that could be intervened upon to halt the cycle of lung injury. This knowledge gap has led to an absence of disease-modifying therapies and a therapeutic pipeline that has thus far led to only incremental advances on existing drugs. The lymphatic vessels are uniquely positioned to regulate inflammatory responses in most tissues because they drain fluid and traffic immune cells in the form of lymph. We have recently published our findings that impaired lymphatic drainage alone is sufficient to induce lung inflammation that culminates in the hallmarks of human COPD. Furthermore, we have discovered that lymphatic endothelial cell injury and lymphatic vessel thrombosis are early events after CS exposure. Interestingly, we not only identified lung lymphatic thrombosis in human lung tissue from patients with COPD, we also found that the degree of lymphatic thrombosis is linked to COPD severity. This work suggests that lymphatic dysfunction may be central to COPD pathogenesis but the role of the lymphatics in CS-induced lung injury and inflammation is not well understood. Filling this critical knowledge gap promises to result in new classes of lymphatic-targeted agents that have the potential to block or reverse COPD and lead to prolonged survival in these patients. We will investigate the novel concept that lymphatic dysfunction is a defining early event in the pathogenesis of emphysema that can contribute to progressive tissue destruction. Therefore, the objective of this proposal is to define the actionable mechanisms that result in lymphatic endotheliopathy after CS exposure and drive progression of COPD. Our central hypothesis is that lymphatic thrombosis due to CS drives disease progression through activation of thrombin in the lymphatic endothelium. Our approach consists of lymphatic- specific manipulation of the coagulation pathway and thrombin signaling in order to mechanistically address the consequences of lymphatic thrombosis on disease progression in a murine model of COPD (Aim 1), and the mechanism by which CS causes lymphatic dysfunction (Aim 2). Furthermore, we will use these models to test how inhibition of thrombin activity or specific inhibition of the thrombin receptor affects the inflammatory response to CS and disease progression. When completed, these studies will lead to a shift in the current paradigm for the pathogenesis of COPD to include early changes in lung lymphatic function. In doing so, we will both broaden the field and set the stage for further studies with immediate clinical relevance given the use of existing therapies to target lymphatic function.

NIH Research Projects · FY 2026 · 2023-04

Type 2 Diabetes (T2D) prevalence increases with age and affects over 13.2% of the US population by 2020. Studies show that microbiota composition and their metabolic genes are different between healthy and T2D patients. However, it is challenging to unravel their causal effects on host glucose homeostasis and T2D due to the lack of an efficient system to manipulate their levels in vivo. Recently, we were able to toggle microbiota amino acid metabolic pathways in vivo and found that some pathways affect host glucose homeostasis. We hypothesize that the gut bacteria impact the host amino acid pool, which will further modulate host glucose homeostasis. Herein we will identify the microbes that actively ferment dietary amino acids and evaluate how they affect host glucose homeostasis in diseased mouse models. We will learn more about the role of microbiota-mediated AA metabolism in the progress of T2D. Our work will also lay the ground for generating a synthetic and engineered gut microbial community with defined metabolic functions to prevent and cure T2D. There are three convergent motivations: First, many microbiota molecular features are different between healthy and T2D patients, and we need functional studies to causally connect them with T2D. We will combine bioinformatics, metabolomics, bacterial genetics, and a gnotobiotic mouse model to modulate microbiome metabolic pathways in the host. This approach will boost a systematic identification of T2D- causing microbiota genes and pathways; our findings will also promote new therapeutic strategies by targeting these previously unknown microbial metabolic avenues. Second, gut microbiota metabolism significantly impacts host metabolic health. However, the molecular mechanisms behind how microbiota regulates host amino acid homeostasis and downstream biology remain largely unexplored. We believe that this approach has huge potential: it can be used to regulate the microbiome metabolic functions at different body sites where host and microbes interact. Our finding would also open the door to interrogating – and ultimately controlling – one of the most concrete contributions that gut bacteria make to host biology. Third, a synthetic microbial community with a defined and programmable metabolic function has therapeutic potential for T2D. The gut microbiome is part of our ‘pan-genome,’ whose metabolic functions are more tractable by genetic manipulation or adjusting microbiota composition. Our approach will expedite the genomic and biological characterization of microbiota metabolic genes, laying the basis for a synthetic community with a defined and programmable metabolic function to prevent and cure T2D and other age-related metabolic diseases.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY The enteric nervous system (ENS) is critical for controlling key intestinal functions such as peristalsis and nutrient absorption. Emerging paradigms indicate that interactions between neurons and immune cells fundamentally impact immunity and inflammation in peripheral tissues. Recently, it has been shown that the ENS can act on the intestinal immune system to modify the function of innate lymphoid cells (ILCs). However, despite these advances, the role of ILCs in ENS function remains unexplored. In my preliminary studies, I defined that ILCs are genetically poised to interact with the ENS and support their development or function. By examining a specific factor produce by ILCs, I defined how it is regulated and developed a lineage-specific knockout to demonstrate this pathway plays a role in augmenting the ENS. To goals of this proposal are, in Aim 1, to utilize in vivo and ex vivo models to determine the molecular mechanisms that regulate ILCs and associated factors, and, in Aim 2, to determine the cellular mechanisms by which ILC-derived factors impacts the ENS and the importance of this interaction in maintaining ENS homeostasis during inflammation. Collectively, these two aims will critically define a novel molecular mechanism by which ILCs impact ENS function in states of intestinal health and disease, which could provoke novel preventative, therapeutic or curative strategies. Further, it will create an outstanding training opportunity for my continued development in science and eventual transition to become an independent academic researcher.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT Although modern antiretroviral (ARV) therapies have dramatically improved the outlooks for people living with HIV, they are unable to cure infection. For people with HIV a cure would represent freedom from many burdens, including stigma, expensive medications, and inflammation-associated co-morbidities. A cure would also have public health benefits, comprising a powerful tool to help end the HIV epidemic. Developing a cure for HIV requires developing an understanding of how the virus persists for years and decades in people, even when new rounds of cellular infection (replication) are blocked by ARVs, and despite the ongoing presence of antiviral immune responses. The dominant paradigm has been that the virus hides in a latent state in infected cells and is thus invisible to immune responses. Efforts to cure infection have therefore focused on therapeutically reversing HIV latency to expose these cells to elimination but have thus far yielded disappointing results. This, along with several converging lines of evidence, have led to more recent hypothesis that hiding from the immune system may not be the only mechanism by which HIV persists – but rather that these rare populations of infected cells may have been selected for those that possess cell-intrinsic resistance to killing by cytotoxic T-lymphocytes, even when they express antigen and are seen. This parallels recent findings from ImmunoOncology where it has now been well established that some immunogenic tumors undergo selection for cell-intrinsic resistance to CTL. For this project, we have assembled a team comprising a pioneer in establishing mechanisms of CTL resistance in tumors, and two HIV experts who have advanced the idea of CTL resistance in this setting through a series of ex vivo studies. By merging these areas of expertise, we Aim to comprehensively describe mechanisms of CTL resistance in HIV-infected primary CD4+ T-cells and to discern which of these are at play in real HIV reservoir cells from people with HIV. We will build from these results to select therapeutic targets and identify combination approaches that integrate these with HIV-specific CTL and latency reversal strategies to achieve specific elimination of HIV reservoir-harboring cells ex vivo. We will also leverage an innovative mouse model to test whether engaging these therapeutic targets limits the seeding of HIV reservoirs in vivo. The results of this project are thus expected to be: i) laying a broad foundation for understanding CTL resistance in the HIV reservoir and ii) pre-clinical validation of multiple therapeutic targets with the potential to contribute to a cure for HIV.

NIH Research Projects · FY 2026 · 2023-03

Modified Abstract Section Congenital cytomegalovirus (cCMV) infection is the leading infectious cause of birth defects and permanent brain damage worldwide, resulting in >5,000 infants each year in the United States alone developing lifelong disabilities. While a vaccine to prevent cCMV has been labeled “tier 1 priority” for over 20 years, we remain without a licensed vaccine product, in part due to limited understanding of the types of immune responses that are protective against placental CMV transmission. Primary infection during pregnancy is high risk for cCMV transmission, yet only approximately a third of mothers acutely-infected during pregnancy will transmit the virus to their infants, suggesting that the rapidity and magnitude of the maternal immune responses plays a role in protection against placental virus transmission. The overarching goal of this proposal is to define CMV specific humoral and cellular immune responses associated with reduced risk of fetal transmission and model their impact on placental transmission. To address this goal, we have access to a unique cohort of 399 acutely CMV-infected transmitting and non-transmitting pregnant women enrolled in the NIH National Institute of Child Health and Human Development (NICHD) Maternal Fetal Medicine Unit (MFMU) CMV hyperimmunoglobulin trial (NCT01376778). This trial was a double-blind randomized trial that screened >100,000 pregnant women for acute CMV infection for enrollment to receive either CMV hyperimmunoglobulin (HIG) or placebo, yet was stopped for futility, creating a unique opportunity to define the acute cellular and humoral immune responses that are associated with transmission risk since HIG infusion after seroconversion did not change transmission risk. Our hypothesis is that the combination of early, functional CMV-specific IgG responses and CD4+ T cell and specialized innate immune cell responses to primary CMV infection during pregnancy will predict reduced risk of fetal transmission and disease. The combined strength of this uniquely large acutely CMV-infected pregnant cohort, our expertise in measuring CMV-specific humoral and cellular immune responses, and expertise in novel mathematical and placental organoid models will inform immune targets of CMV vaccine development that will be predicted to reduce the risk of cCMV transmission. Our Specific Aims include: 1) Define the CMV-specific IgG binding and functional responses associated with reduced transmission and disease following primary CMV infection in pregnancy; 2) Define the cellular immune responses elicited during primary CMV infection that associate with reduced transmission in pregnancy; 3) Develop an in silico model that can predict candidate CMV vaccine efficacy for prevention of placental transmission based on maternal immune correlates of cCMV transmission and the rate of viral spread in placental organoid models. Defining immune targets that will reduce fetal transmission and infant disease following primary maternal CMV infection will speed the design of effective vaccines to drastically decrease neurologic impairment and long-term disabilities in all children worldwide.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Among aggressive lymphomas, the Diffuse large B-cell lymphomas (DLBCLs) are among the most highly proliferative, and have massive requirements for production of metabolic precursors. Along these lines we recently identified NAD+ dependent lysine deacetylase SIRT3, as a master regulator of mitochondrial stress metabolism, as a critical driver of DLBCL growth and survival. We showed that SIRT3 loss of function in DLBCL cells kills lymphomas by disrupting their ability to use glutamine and other amino acids in the TCA cycle, which triggers destructive autophagy – both in vitro and in vivo. Importantly, we created the mitochondrial targeted SIRT3 selective small molecule YC8-02 that precisely mimics the effect of SIRT3 depletion and potently killed DLBCL cells in vitro and in vivo. These compounds yielded further enhanced killing effects in combination with targeted therapies such as venetoclax as well with chemotherapy drugs commonly used to treat DLBCLs. Finally, our mechanistic data point to pathways in cells that could eventually lead to resistance to SIRT3 targeted therapy, as well as ways to prevent this from happening so as to yield maximal therapeutic efficacy. An overarching challenge in delivering precision medicine for DLBCL patients is their marked genetic heterogeneity and extreme abundance of somatic mutations. There are currently no targeted agents with activity and targets relevant to more than a small fraction of patients. However, we identified SIRT3 as a broadly relevant and critical non-oncogene addiction that is required by DLBCLs independent of their genetic background. Through this proposal we provide the basis for i) translating SIRT3 inhibitors to the clinic, ii) understanding and mitigating potential resistance mechanisms, and iii) incorporating SIRT3 inhibitors into rationally designed anti-lymphoma regimens with broad relevance for the subsets of patients who desperately need improved therapies. Our proposal uses state of the art model systems physiologically relevant to these complex tumors, and is poised to deliver highly impactful outcomes from the scientific and clinical perspective.

NIH Research Projects · FY 2026 · 2023-03

Project Summary/Abstract Stroke is a prevalent and devastating disease with limited therapeutic options. Inflammation and immune cells are major components in the pathophysiology of ischemic stroke and contribute to acute and delayed tissue injury. However, our incomplete understanding of the factors regulating the immune responses triggered by cerebral ischemia remains a significant obstacle to the development of effective therapeutic interventions based on modulating post-ischemic inflammation. Besides activation of brain resident immune cells, ischemic stroke is characterized by the recruitment of peripheral innate and adaptive immune cells that participate in the inflammatory response and contribute to the damage. Commensal microbiota that populate epithelial surfaces play a defining role in shaping the immune system, the development, maintenance and function of which depends critically on the relative abundance and composition of the different microbial species. In particular, intestinal commensal bacteria, the most abundant symbiotic compartment in the body, have emerged as a potent regulator of the immune response to stroke. The long-term goal of this research program is to elucidate the role of intestinal microbiota in stroke pathobiology and develop the experimental framework for new preventative and therapeutic approaches for ischemic stroke. In the present application, we will test the hypothesis that the interaction of commensal intestinal microbiota with dendritic cells is a critical determinant of stroke outcome by modulating the immune system and inflammatory response to cerebral ischemia. Supported by relevant preliminary results, this application will test the hypothesis that commensal intestinal microbiota modulate stroke outcome by acting on intestinal dendritic cells to either induce a tolerogenic or pro-inflammatory phenotype affecting T cell differentiation. These intestinal immune changes propagate to the brain and meninges after stroke by increased immune cell trafficking. To this end, we will determine (a) the roles of different dendritic cell populations in mediating the neuroprotective effects of altered microbiota and (b) the cellular and molecular targets of microbiota that lead to altered intestinal immunity and their importance for stroke outcome. These goals will be achieved using a mouse model of transient focal cerebral ischemia with assessment of histological and neurological outcome, a model of altered gut bacteria, cell tracking of intestinal immune cell, and in vitro immune cell coculture models. This proposal may open the way to new avenues for stroke prevention and therapy based on modulation of the immune system by the gut microbiota.

NIH Research Projects · FY 2026 · 2023-03

Clinical analyses show that UBR5 gene amplifications and overexpression occur in 10-40% cases of many major types of aggressive human cancers. Furthermore, breast, ovarian and prostate cancer patients carrying genetic alterations in UBR5 have significantly reduced survivals compared to those without the lesions. Our experimental work in vitro and in vivo has first demonstrated that UBR5, functioning like an “oncogene”, plays a profound role in promoting breast and ovarian cancer growth and metastasis. We have also shown that tumor-derived UBR5 drives malignant triple negative mammary tumor growth through both cell-intrinsic and extrinsic mechanisms, whereas it facilitates metastasis primarily in a tumor cell-autonomous manner. Thus, further elucidating UBR5’s fundamental biology and identifying critical signaling nodules controlled by UBR5 in its potent tumorigenic and immunoregulatory activities will not only advance the science but also help the development of novel therapies for highly malignant breast cancer that evades the endogenous cellular control mechanisms and resist current interventional strategies. We hypothesize that UBR5 promotes aggressive BC/TNBC via distinct mechanisms that include controlling the CDC73 protein turnover in an E3 ubiquitin ligase-dependent manner and enhancing Interferon-g-induced transcription of the PDL1 gene and others in an E3 ligase- independent manner. We propose to broaden and expand the exploration of the cellular and molecular mechanisms of these modulations in two major specific aims. (1) To characterize the biochemical basis of CDC73 protein regulation by UBR5 acting as an E3 ubiquitin ligase; and investigate the role of the chemokine CXCL16 in mediating CDC73’s immunostimulatory activities via recruitment of cytotoxic T lymphocytes to the tumor site. (2) To investigate the cellular and molecular mechanism whereby UBR5 broadly enhances the IFN--activated signaling pathway independently of the E3 ligase activity and explore the therapeutic potential of pharmacological UBR5 inhibition. The outcome of these studies will pave the way for developing innovative therapeutic strategies for highly aggressive and therapy-resistant breast cancer by targeting UBR5 and/or its crucial signaling pathways.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY Difficulty initiating or executing appropriate movement is characteristic of neurological disorders including Parkinson’s disease and ataxia, while the production of abnormally repetitive movements is seen in neurological and neuropsychiatric disorders such as Tourette Syndrome, OCD and ASD This research is aimed at understanding the neuronal mechanisms by which C. elegans nematodes initiate, execute and stabilize appropriate motor actions, in order to make predictions about how dysregulation of motor action patterns arise. Currently, our understanding of the mechanisms that generate appropriate motor outputs in physiological states and abnormal outputs in pathological states is incomplete. With 302 neurons with known connectivity and numerous genetic tools to target and manipulate individual neurons, the nematode Caenorhabditis elegans offers an excellent system to study the neuronal mechanisms of motor output generation. C. elegans locomotion is composed of a stable of sequence of motor actions/states: from forward locomotion to reversal with or without a turn, then a resumption of forward locomotion. Past studies have associated the C. elegans interneurons AIB, RIM, and AVA with reversals, however the exact neuronal contributions required to initialize, execute and stabilize motor states remain elusive. Based on their connectivity and previous experimental results, I hypothesize that AIB, RIM and AVA primarily initialize, stabilize and execute reversals, respectively, and that these functions will be reflected in their response to optogenetic perturbation, their required temporal windows to drive motor state changes and their response to combinatorial perturbation. In Aim 1, I will express the excitatory optogenetic channel, Chrimson, or inhibitory optogenetic channel, GtACR2, individually in single neurons to understand the state-dependent timing of single neuron activation or deactivation that drives motor state changes. In Aim 2, I will use the bidirectional optogenetic tool BIPOLEs (a Chrimson and a GtACR2 channel in tandem) to determine the precise temporal windows of activity required for reversal-associated interneurons to produce expected motor output. In Aim 3, I will combine optogenetic perturbation with chemogenetic silencing in order to understand the interactions between neurons required to generate stable, flexible motor states. Dissecting motor output changes in C. elegans may elucidate broader themes in motor pattern generation and its dysregulation. This research will take place in a highly supportive, inter-disciplinary laboratory environment. It requires the use of novel genetic tools for neural circuit perturbation and computational behavioral analysis, ideal for my training as a future physician-scientist studying the genetic and circuit mechanisms of behavior in health and disease.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Up to 60% of older persons with Alzheimer’s disease and related dementias (ADRD) suffer from bothersome pain and nearly half experience pain-related activity limitations. Despite best-practice guidelines calling for routine pain assessment of persons with ADRD, pain is severely under-detected and poorly managed in this population. A major barrier to the identification and treatment of pain in persons with ADRD is impaired communication. As ADRD progresses, neurodegenerative changes impede individuals’ ability to understand and verbally articulate their discomfort. In such cases, reliable self-reports of pain are not feasible and behavioral assessment is recommended. Informal (family) caregivers are well situated to detect pain and facilitate management in persons with ADRD, given their extensive involvement in care activities. However, caregivers receive virtually no guidance or training in these areas. To address the challenges that ADRD caregivers face in recognizing and communicating about pain, the PI (Riffin) and her interdisciplinary team of Co-Investigators developed a manualized, multicomponent intervention, the Pain Identification and Communication Toolkit (PICT). PICT is informed by theories of behavior change and pain communication and includes (a) training in administering an observational pain assessment tool, (b) coaching in effective pain communication, and (c) building caregivers’ skills through routine practice. The team’s NIA-funded pilot trial with a racially and ethnically diverse group of caregivers (14% Black, 15% Hispanic, 8% multiracial) demonstrated the feasibility and acceptability of PICT, and preliminary impact on caregivers’ communication with healthcare providers. The proposed R01 builds on this prior research by using an Experimental Medicine approach, grounded in the Science of Behavior Change, to evaluate PICT’s efficacy, mechanisms of action, and potential moderators in a Stage II clinical trial. It will leverage the infrastructure of a community-based Managed Long-Term Care (MLTC) program in New York with wide socioeconomic and racial diversity to (1) determine the efficacy of PICT on caregivers’ pain recognition and communication (Primary Outcomes), caregivers’ distress and burden; patients’ physical function, behavioral disturbance, changes in pain treatments or regimens, and institutionalization (Secondary Outcomes), (2) identify the patient and caregiver factors that may moderate the effects of PICT on study outcomes, and (3) evaluate the mechanisms (theoretically-derived variables) by which PICT affects study outcomes. Overall, this research represents a critical step toward addressing the under-detection and under-management of pain in persons with ADRD, supporting the largely hidden but vital ADRD caregiver workforce, and laying the groundwork for a future multisite pragmatic trial.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY AND ABSTRACT People living with HIV (PLWH) in rural sub-Saharan Africa are three times less likely to achieve viral suppression than their urban counterparts. Novel HIV service delivery models for rural PLWH are needed to improve the HIV continuum of care and achieve viral suppression. Traditional healers (TH) are lay providers who serve as the first line of healthcare in rural Africa, and frequently provide care to PLWH who have disengaged from HIV care. TH are accessible, trusted members of rural communities, but have not been integrated into HIV care programs. Our prior cluster randomized trial demonstrated that partnerships with TH quadrupled the uptake of HIV testing in rural Uganda through facilitation of HIV counseling and self-testing. Building on these results, we adapted an evidence-based lay provider intervention for delivery by Ugandan TH to support subsequent steps of the HIV continuum. The TH-delivered program is called Omuyambi (“Support” in Runyankole) and includes assisting PLWH to link to care for ART initiation, providing ongoing counseling on ART adherence, and encouraging retention in clinical care. We conducted a pilot study of this lay provider program among 12 TH and 20 PLWH who were disengaged from HIV care or ART naïve. Results were overwhelmingly positive: 100% of PLWH linked to HIV care and initiated ART within 14 days, 95% reported ART adherence and 100% were retained in care after nine months. Building upon this evidence, we hypothesize that TH can support clinic-based care and improve viral suppression among rural PLWH. We will conduct a hybrid type I effectiveness-implementation cluster randomized trial to evaluate the effectiveness of the Omuyambi intervention on viral suppression among ART naïve/defaulted PLWH in Uganda. • Aim 1: Compare the Omuyambi intervention versus routine HIV clinic-based care (control) in a cluster randomized trial. Forty TH clusters that include ≥650 PLWH will be randomized to the Omuyambi intervention or to a control arm, in which TH will refer PLWH to clinic-based HIV care alone. Primary clinical outcome is viral suppression at 12 months measured via dried blood spot analysis. We hypothesize that 80% of PLWH in the intervention arm will achieve viral suppression, compared with 60% in the control arm. • Aim 2: Evaluate implementation of Omuyambi using a convergent mixed methods study design and the Consolidated Framework for Implementation Research (CFIR). Qualitative and quantitative data will be collected from participating TH, PLWH, HIV clinic staff, and Ministry of Health Officials. These data will be used to assess Omuyambi implementation determinants and outcomes. The proposed research is significant as it responds to the World Health Organization and Ugandan Ministry of Health calls for community-based interventions to improve HIV viral suppression where current programs have suboptimal impact. If effective, this approach has the potential to improve the HIV continuum among rural PLWH towards UNAIDS 95-95-95 benchmarks necessary to end the HIV epidemic.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Asthma results from a complex interplay between genetic background and environmental triggers. The 17q21 asthma susceptibility locus is strongly linked to childhood asthma through ORMDL3. ORMDL3 regulates serine-palmitoyl CoA transferase (SPT), the critical enzyme for the de novo synthesis of sphingolipids. We demonstrated that decreased SPT activity leads to airway hyperreactivity. There has since been increasing evidence that sphingolipid metabolism is altered in airway epithelial cells and animal models of ORMDL3-associated asthma. We have recently shown that children with asthma have decreased sphingolipid synthesis, especially in the presence of asthma risk 17q21 genotypes. 17q21 genotypes are also linked to the risk of developing asthma following respiratory infections with human rhinovirus (HRV). This is relevant as HRV is also the most common trigger for asthma attacks. Supported by preliminary data in mice and airway epithelial cells demonstrating strong similarities in sphingolipid levels and gene expression between HRV infection and sphingolipid deficiency, we hypothesize that HRV infection can further impair sphingolipid synthesis. We propose to study the effects of HRV on sphingolipid synthesis in children with asthma and airway epithelial cells. These studies could then further solidify the central role of sphingolipids in asthma pathogenesis that has been predicted by the commonality and the strong association of 17q21 genotypes to asthma. Two specific aims are proposed to assess the overall hypothesis that the sphingolipid de novo synthesis pathway is critical not only for asthma pathogenesis but also in response to the most common trigger for asthma attacks. In Aim 1, we will determine sphingolipid synthesis in the respiratory tract in children with asthma and HRV infection. Sphingolipids will be assessed in nasal fluid, blood, and gene expression in nasal cells obtained from children during and after the resolution of HRV-triggered asthma attacks. In a subaim, we will evaluate the effects of HRV on sphingolipid metabolism and gene expression in primary human airway epithelial cells (HAEC) with homozygous for a common 17q21 asthma variation that leads to decreased blood sphingolipids in children with asthma. HAEC from an established biorepository from adult donors and nasal brushings from children all grown at air-liquid interface will be infected with HRV and RSV and evaluated for effects on sphingolipid synthesis. For Aim 2, we will test the hypothesis that the altered ratio of the sphingolipid mediator sphingosine 1-phosphate and sphinganine 1 phosphate, which we found in SPT deficiency and children with the 17q21 asthma risk genotypes, leads to airway hyperreactivity. Overall, these studies not only inform on the role of sphingolipids in the pathogenesis of asthma and the relation to its most common trigger but may lead to new therapeutic approaches involving sphingolipid metabolism.

NIH Research Projects · FY 2026 · 2023-02

ABSTRACT Clinical trials are often conducted under idealized and rigorously controlled conditions to ensure internal validity (maximizing potential treatment efficacy) while balancing patient safety (e.g., serious adverse events [SAEs]); but these conditions—often reflected in trials’ eligibility criteria—paradoxically, limits (1) the ability to identify the “right” study populations of the trials, and (2) the trials’ generalizability to the target population in real-world settings. Low generalizability has long been a concern, including for Alzheimer's disease (AD) trials. AD trial participants are systematically younger than AD patients in the general population, where eligibility criteria design issues are arguably the biggest yet modifiable barriers. The FDA has launched numerous initiatives to improve trial design and enrollment practices, such as using enrichment strategies (e.g., “use patient characteristic to select a study population in which detection of a drug effect [or safety event] is more likely than it would be in an unselected population”), so that the trial participants can better reflect the real-world target population and the trials are more likely to succeed. However, there are significant gaps between the need to improve AD trial eligibility criteria design and ways available to fulfill the need in practice. On the other hand, rapid adoption of electronic health record (EHR) systems has made large collections of real-world data (RWD) that reflect the characteristics and outcomes of the patients being treated in real-world settings, available for research. The increasing availability of RWD combined with the advancements in artificial intelligence (AI), especially machine learning (ML) offer untapped opportunities to generate real-world evidence (RWE) to support eligibility criteria design for AD trials, due to a number of key methodological gaps: (1) the lack of validated computable phenotyping (CP) and natural language processing (NLP) algorithms and tools that can accurately define the populations (e.g., AD patients) of interest and extract key relevant patient characteristics and outcomes of interest (e.g., trial endpoints such as MoCA and safety profile such as SAEs) from RWD, (2) the lack of ways to identify the desired study populations (and corresponding eligibility criteria), considering the impact of criteria to potential treatment effectiveness, patient safety, and study generalizability, and (3) the need of an easy-to-use toolbox to support trialists’ eligibility criteria design process. We propose (1) novel causal- principled, explainable AI (XAI) approaches to generate RWE to facilitate AD trial eligibility criteria design, and (2) to create the web-based ALZHEIMER'S DISEASE ELIGIBILITY EXPLAINER (ADEP) tool. We will leverage two large RWD resources, the OneFlorida+ (~19 million patients from Florida, Georgia, and Alabama) and INSIGHT (~12 million New Yorkers) clinical research networks (CRNs) contributing to the national Patient-Centered Clinical Research Network (PCORnet). The success of this project will establish (1) a novel, generalizable, and XAI-based trial enrichment framework with large collections of distributed RWD, and (2) a prototype toolbox that can provide RWE to eligibility criteria design, balancing effectiveness and patient safety in the target population.

NIH Research Projects · FY 2026 · 2023-02

Immune checkpoint inhibitors (ICI) targeting the PD-1/PD-L1 axis have improved survival in non-small cell lung cancer (NSCLC). However, most patients fail to achieve durable benefit due to persistent immunosuppressive barriers in the tumor microenvironment (TME). Stereotactic body radiation Therapy (SBRT) enhances responses to ICI in preclinical and clinical settings, but the cellular mechanisms mediating this interaction are not fully defined. Our prior work demonstrated that low dose SBRT in combination with ICI significantly reduces tumor growth and improves survival in murine NSCLC models. Unexpectedly, this therapeutic benefit required lung resident Scgb1a1+ club cells; genetic ablation of club cells eliminated radiation-induced enhancement of cytotoxic T cell activity and survival benefit (Ban et al, Nature Cancer, 2021). These findings led to the hypothesis that radiation activates club cells to produce immunostimulatory factors that reprogram the TME and potentiate ICI efficacy. Our recent mechanistic studies showed that irradiation of immortalized murine club cells (C22) induced G2/M arrest and activated inflammatory transcriptional programs, including interferon-α response, TNFα/NF-κB signaling, and type I interferon pathways. Quantitative RT-qPCR confirmed upregulation of canonical interferon-stimulated genes (Isg15, Stat1, Oasl2, Il6, Ptgs2) and Ifnb1 key factors in Type 1 Interferon response pathway. In vivo, lung irradiation (3 × 4 Gy) increased IFNβ expression specifically in club cells, as demonstrated in IFNβ-YFP reporter mice. Pharmacologic blockade of type I interferon signaling with anti-IFNAR1 antibody abrogates the therapeutic efficacy of SBRT in combination with ICI, establishing a functional requirement for this pathway.   We hypothesize that radiation-induced DNA damage in club cells activates innate sensing pathways (e.g., cGAS-STING), leading to type I interferon production that enhances antigen presentation, dendritic cell activation, and cytotoxic T cell priming. Aim 1 will define the molecular pathways by which irradiated club cells reprogram the tumor immune microenvironment to generate durable anti-tumor immunity. Aim 2 will leverage specimens from an ongoing ICI/SBRT clinical trial to determine whether club cell-associated interferon signatures and related biomarkers correlate with therapeutic response and metastatic recurrence in human NSCLC. By identifying club cells as a previously unrecognized mediator of radiation-induced immune priming, these studies will define a non-tumor epithelial mechanism of SBRT-ICI synergy and provide biomarker-driven refinement of SBRT-ICI combinations in NSCLC.

NIH Research Projects · FY 2026 · 2023-02

Project Summary Intravenous lipid emulsions (ILEs) are important therapies. Though designed for parenteral nutrition, they were discovered to be an antidote for local anesthetic systemic toxicity (LAST). With rising intravascular concentrations of local anesthetics (LAs), which canonically inhibit voltage-gated sodium channels (Nav), symptoms of central nervous system (CNS) toxicity progress to seizures, and cardiac arrest may ensue. LAST is a major clinical challenge, and the identification of ILEs as treatment was a breakthrough that has made use of LAs safer. Despite this important role, the molecular mechanisms by which ILEs treat LAST are not fully understood. Furthermore, there is a paucity of studies examining whether Intralipid®, the emulsion recommended in medical guidelines that is composed of long-chain triglycerides from soybean oil, is the best choice. The most simplistic model for the effects of ILEs is that they form an intravascular compartment to partition LAs and lower the effect-site concentration, but the degree to which concentrations fall in the aqueous phase is debated. A competing hypothesis with strong evidence in cardiac models proposes ILEs serve as a fuel via β-oxidation of triglycerides (TGs), overcoming mitochondrial dysfunction caused by LAs. This mechanism has been dismissed to explain reversal of CNS dysfunction because of the dogmatic belief that neurons rely on glucose metabolism. Additionally, neurotoxicity due to LAs has been attributed, with limited evidence, to preferential block of inhibitory neurons, but important effects in the CNS are unexplained by this hypothesis. My preliminary in vitro data shows that neurons can use Intralipid® to sustain synaptic function in the absence of other fuels, supporting the metabolic hypothesis of ILEs for LAST in the CNS. This research proposal will test the hypothesis that ILEs reverse neurotoxicity by overcoming LA-induced mitochondrial dysfunction. The experimental plan will systematically investigate the capacity for neurons to metabolize components of ILEs. In doing so, I will comprehensively investigate the suitability of lipids to fuel synaptic function. In Aim 1, I will use my developing expertise in optical imaging of cultured neurons expressing genetically-encoded biosensors to compare lipids by their ability to sustain synaptic vesicle recycling and produce ATP when deprived of glucose. Lipids will be tested as emulsions of both long- and medium-chain TGs and free fatty acids of different lengths and saturation. In Aim 2, a metabolically optimized emulsion will be compared in culture to Intralipid® in its ability to reverse LA-induced synaptic dysfunction. In Aim 3, the optimized emulsion will be compared to Intralipid® in mice, quantifying effects on seizures with widefield calcium imaging, local field potentials, and autofluorescence flavin imaging of metabolic activity. My five-year research and career proposals capitalize on my excellent mentors and institutional environment. I will acquire the publication record and expertise necessary for recognition as a national leader in anesthetic neuropharmacology and prepare for independent investigation with R01 funding.

NIH Research Projects · FY 2026 · 2023-02

Abstract FDA approved molecularly targeted therapies have not significantly reduced mortality in non-small cell lung cancer (NSCLC) patients. Therefore, identification and characterization of novel targets for developing effective therapies is warranted. In this proposal, we will develop endoplasmic reticulum (ER) sensor IRE1 as a potential therapeutic target in NSCLC. Adverse conditions in the tumor microenvironment (TME) can rapidly disrupt the protein folding capacity of the ER, thereby triggering a state of cellular “ER stress”. The ER stress response arm of the unfolded protein response, particularly the conserved IRE1-XBP1 pathway has emerged as a central orchestrator of malignant progression. Activated during periods of ER stress, the IRE1 RNAse domain cleaves its downstream target X-box binding protein (Xbp1) mRNA (inactive, XBP1u), converting it to active isoform (XBP1s), which serves as a functionally active transcription factor. We have determined that increased expression of XBP1s is associated with poor survival in NSCLC, cancer cell-intrinsic deletion of IRE1 delayed malignant progression and extended survival in mouse models of NSCLC. IRE1 deficiency triggered protective type-I IFN responses associated with marked reprogramming of both the lymphoid and myeloid cell subsets. These findings have led to the hypothesis that dysregulated IRE1-XBP1s signaling in cancer cells facilitates NSCLC progression by governing key immunomodulatory programs in the tumor microenvironment (TME). Therefore, understanding the underlying mechanisms has the potential to generate unique therapeutic strategies against difficult to treat NSCLC. We will determine mechanisms by which persistent activation of ER stress sensor IRE1 drives immunosuppression in the TME (Aim 1), determine if pharmacological inhibition of IRE1α sensitizes NSCLC to PARP inhibitors, and STING agonists by enhancing type-I IFN responses (Aim 2), and assess the clinical relevance of a novel IRE1 gene signature in predicting treatment outcomes in human NSCLC (Aim 3). This proposal is conceptually and technically innovative as it seeks to investigate for the first time how cancer cell-intrinsic IRE1α activation drives immunosuppression in the TME that facilitates immune evasion by disrupting cDC1 function and blunting type-I IFN responses. We expect that the mechanistic insights from these studies will generate unique translational opportunities that may lead to the design of future clinical trials with clinical grade IRE1 inhibitors.

NIH Research Projects · FY 2026 · 2023-01

PROPOSAL SUMMARY Major histocompatibility complex class I (MHC-I) molecules present peptides at the cell surface to CD8 T cells. The transporter associated with antigen processing (TAP) is a heterodimeric molecule of TAP1 and TAP2 that lies at the center of a macromolecular peptide loading complex tasked with loading and folding MHC-I molecules with peptides. TAP shuttles cytosolic proteasome-generated peptides across the membrane of the endoplasmic reticulum (ER) for luminal delivery and loading of MHC-I molecules. Given the crucial role of TAP in translocating peptides to MHC-I molecules, many clinically important human viruses such as Herpesviridae and Poxviridae have evolved strategies to block TAP and evade host CD8 T cell recognition. TAP blockade upon infection of dendritic cells (DC), which are responsible for naïve CD8 T cell priming, impairs conventional TAP-proteasome processing for the classic MHC-I presentation of peptides to CD8 T cells. In fact, the current paradigm holds that TAP blockade in DC renders these cells non-functional and incapable of priming a CD8 T cell response. Priming virus-specific CD8 T cells falls on uninfected TAP-sufficient bystander DC through cross-presentation, a process that enables MHC-I loading with viral peptides derived from DC internalized virus-infected dying cells. However, CD8 T cells primed by TAP-sufficient DC recognize dominant TAP-dependent peptides, whose presentation is severely reduced on tissues infected with immune evasive viruses. TAP-dependent CD8 T cells would also be mismatched to the TAP-independent peptides liberated by alternative TAP-independent processing of viral antigens and presented by MHC-I on those infected tissues. Either scenario creates a diminished or mismatched CD8 T cell target. How does the immune system get around this problem? We found that DC without functional TAP rely on cell-autonomous delivery of MHC-I from a new location, the ER-Golgi intermediate compartment (ERGIC), to internalized antigens to rescue MHC-I presentation and nevertheless cross-prime CD8 T cells. We call this pathway non-canonical cross-presentation. Our findings point to non-canonical cross-presentation as a previously unrecognized pathway for priming CD8 T cells that recognize TAP-independent epitopes and would be best-matched against immune evasive viruses. Studying non-canonical cross-presentation is important to understand the full spectrum of CD8 T cells that can be mobilized against infection, especially if such T cells provide potent local cross-protection within infected tissues. We seek to understand the role of non-canonical cross-presentation in priming a TAP-independent CD8 T cell response against viral infection. Using novel models, we will identify DC that conduct non-canonical cross-presentation, and define the repertoire of TAP-independent epitopes they present to antigen-specific TAP-independent CD8 T cells. We will create novel tools to track TAP- independent CD8 T cell responses and determine whether non-canonical cross-presentation can drive cross- protective immunity against viral variants and immune evasive viruses. Understanding non-canonical cross- presentation will inform universal vaccine design and new therapies for chronic and persistent viral infections.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Dr. Lindsey Reif is a global health epidemiologist with a career goal to leverage epidemiologic research to implement high-impact health interventions for adolescents and young adults living with HIV (AYALH). Over the past decade, she has published 10 first-author papers focused on evaluating health outcomes and interventions among AYALH in Haiti. AYALH, who have survived decades of HIV, now face HIV-related comorbidities, specifically cardiovascular disease (CVD), as they age into adulthood. Her K01 objective is to expand her training and research to focus on HIV-related preclinical CVD— asymptomatic cardiometabolic abnormalities indicating the onset of CVD—among AYALH. She will adapt evidence-based interventions to prevent early onset CVD with impact for AYALH across the lifecourse. Her training objectives are to build expertise in: 1) CVD epidemiology among AYALH, 2) mixed-methods, and 3) implementation science in order to design and implement interventions to prevent CVD morbidity and mortality. Her career development plan includes multidisciplinary mentorship from a team that brings decades of experience in research among AYALH, HIV and CVD epidemiology, and implementation science across global health settings. The team has an impeccable track record of mentorship in global health research. Dr. Reif will also take complementary advanced coursework to support her training objectives. Her K01 is supported by a robust research and training environment that leverages a 40-year collaboration between Weill Cornell (New York) and GHESKIO (Haiti). Dr. Reif’s research plan establishes the epidemiology of preclinical CVD among AYALH and adapts an evidence-based intervention to prevent the onset of preclinical CVD and its progression. She will conduct a prospective cohort study among 500 HIV+ and 500 HIV- participants ages 18-30 years, followed for 3 years in Haiti. In Aim 1, she will evaluate the prevalence, age of onset and progression of preclinical CVD (pre- hypertension, microalbuminuria, and dyslipidemia) by HIV serostatus. She will evaluate risk factors in 3 domains: lifestyle, HIV-related, and psychosocial. She hypothesizes clusters of risk factors, driven by HIV- related and psychosocial factors, are associated with preclinical CVD among AYALH. In Aim 2, she will adapt the evidence-based WHO HEARTS technical package for AYALH in LICs using mixed methods and implementation science. This K01 addresses the knowledge gap of the epidemiology of preclinical CVD among AYALH and informs the adaptation of evidence-based CVD interventions to prevent premature disability and death among this population as they emerge into adulthood. Dr. Reif’s training and career development plan complement and expand her existing skillset and culminate in the submission of an NIH R01 facilitating her transition to independent investigator.