Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Dementia family caregivers frequently experience persistent musculoskeletal pain. Pain is often made worse by performing caregiving tasks, impairing dementia family caregivers’ ability to provide care. Dementia family caregivers’ who have pain often have co-existing emotional and psychological health challenges, such as high negative emotions and/or comorbid depression, which makes managing their pain and their caregiving tasks more difficult. The objective of this K99/R00 proposal is to develop Dr. Shelbie Turner’s capacity to conduct translational research to support dementia family caregivers’ health and well-being through improved management of their own persistent pain. Dr. Turner will work towards five training objectives in the K99 phase of the project. Doing so will allow her to develop and transition to an independent investigator who designs, adapts, evaluates, and disseminates innovative, evidence-based interventions to prevent or mitigate dementia family caregivers’ negative health outcomes, with a focus on the problem of persistent pain and associated symptoms (e.g., negative emotions) and comorbidities (e.g., depression). First, she will add to and extend her knowledge in core substantive areas to include pain, negative emotions, dementia caregiving, and health behavior change. Second, she will improve her ability to develop, implement, and disseminate behavioral interventions with an emphasis on both pain management interventions and caregiver interventions. Third, she will gain experience needed to design and conduct clinical trials, including conventional, pragmatic, and adaptive clinical trials with dementia family caregivers. Finally, she will pursue professional development opportunities in grant writing (fourth training objective) and leadership and collaboration (fifth training objective). The three research aims of this application will progress as follows. Aim 1 adapts an evidence- based pain self-management intervention, Problem Adaptation Therapy for Pain in Primary Care (PATH-Pain), for use by dementia family caregivers with persistent pain. Aim 2 tests the usability of the adapted intervention with dementia family caregivers, and Aim 3 examines the feasibility, acceptability, and preliminary efficacy of the intervention through a pilot randomized controlled trial. The adapted intervention will be the first evidence- based pain self-management program customized specifically for dementia family caregivers. The proposed project is consistent with NIA’s mission to conduct behavioral research on aging and foster the development of research scientists in aging. It is aligned with NIA’s special interests in dementia caregiver intervention research and pain management research. Dr. Turner proposes to pursue her developmental goals and begin the proposed research with the support of the Division of Geriatrics and Palliative Medicine at Weill Cornell Medical College of Cornell University, which provides an ideal environment of research support and resources to help her achieve her training and research goals.

NIH Research Projects · FY 2026 · 2024-08

Summary The development of the human body involves cell specification and cell fate transition starting from the embryos. Precise coordination of gene expression networks is required: lineage-specific genes are transcriptional activated during early development, while genes for ectopic lineages are repressed in the process. The precise control of gene expression is governed by epigenetic chromatin modifications. Recent advances showed that genes encoding epigenetic “writer” and “eraser” enzymes are frequently mutated in human diseases. However, treatment or early prevention methods are hampered by the lack of knowledge on how the epigenetic landscape is precisely regulated. We are investigating how a critical histone methylation (trimethylation of lysine 27 on histone H3, or H3K27me3) is precisely regulated. H3K27me3 is the hallmark for facultative heterochromatin, which dynamically regulate gene repression during body development. During early differentiation, H3K27me3 is deposited on pluripotency genes, erased on cardiac genes, and maintained on other developmental genes for ectopic lineages. The dynamic level of H3K27me3 across the genome is essential for the ON/OFF switch of gene expression, but it remained unclear how H3K27me3 is regulated in a coordinated temporal and spatial manner. We hypothesize that key macromolecular interactions including protein-protein and protein-nucleic acid interactions regulates the specificity of the H3K27me3 “writer” enzyme – Polycomb Repressive Complex 2 or PRC2. Recent progress and our preliminary data show that the dynamic interactions between PRC2 and its accessory proteins play key roles in the spatiotemporal regulation of epigenetic silencing specificity. To test the hypothesis, we propose to fully interrogate the mechanism by employing a series of separation-of- function mutants. Understanding the mechanism will open to door to further identification of novel therapeutical targets to manipulate gene expression through epigenetic mechanisms.

NIH Research Projects · FY 2026 · 2024-08

Deforestation is a critical global issue that leads to extensive macro-level changes in our environment. While past literature has linked sudden or expansive environmental changes to substance use, a knowledge gap exists on how deforestation may influence tobacco and alcohol consumption. The objective of this K99/R00 is to establish clearer linkages and quantify effects and associations between deforestation and tobacco and alcohol use, using Indonesia as the geographic focal area. The K99 phase is designed to achieve this objective and augment the candidate’s prior research experience by providing training in: (a) the determinants and epidemiology of substance use and mental health, (b) satellite remote sensing and image classification, (c) advanced methods for environmental epidemiology, and (d) causal designs and software for reproducibility and accessibility. This training is critical to the candidate’s long-term goal of becoming a leading epidemiologist who builds and uses rigorous statistical approaches to examine environmental exposures and social and behavioral health outcomes in the United States and in low- and middle-income countries. The proposed research will draw upon satellite imagery to detect changes in forest cover and composition and 10 years of data (2012-2022) from a nationally and sub-nationally representative, repeat cross-sectional survey. Aim 1 (K99 phase) will examine associations between forms of deforestation (wildfire and logging) and tobacco and alcohol use in the general population and among subgroups. It will subsequently estimate tobacco and alcohol use under different forest loss scenarios to estimate what would be the population-weighted burden of use attributable to incremental increases in deforestation, compared to the counterfactual of stably forested areas. Aim 2 (R00 phase) will develop a new causal inference method for estimating the “pure” effect of deforestation on tobacco and alcohol use. Economic incentives from palm oil production introduce unmeasured confounding and lead to differential exposure probability when illegal wildfires are set to clear land. A longitudinal extension of differential comparison design, also known as “negative or secondary controls,” will be created in a simulation study to contrast three groups (wildfire zones with and without palm oil plantations and stably forested zones) and parse the “pure” effect of deforestation. The new method will then be applied to the real data for estimation. A software package and tutorial will be developed to facilitate use and accessibility. Aim 3 (R00 phase) will examine which combination of factors clustered within regencies are most important in tobacco and alcohol use and model joint effects in probit Bayesian kernel machine regression. The proposed research aligns with NIEHS’ Mission, Vision, and Strategic Plan.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ ABSTRACT Aging, the gradual decline in an organism's physiological functions over its lifespan, impacts all individuals. Yet, why do some individuals age faster and show a greater susceptibility to age-related diseases, including cancer, tissue fibrosis, and diabetes, compared to others? Mounting evidence supports the idea that aging, and the associated process of cell senescence, are rooted in metabolic dysfunction. Metabolic dysfunction occurs when chemical reactions become dysregulated during aging and negatively alter the body's processing and distribution of nutrients, inducing the aberrant accumulation of certain metabolites. These metabolites can affect cellular function by altering signaling processes and protein modifications, contributing to aging progression. The link between aging and metabolism has solidified itself as one of the new challenges in understanding the complexity of aging as a systemic disease. Identifying metabolic pathways and metabolites that drive cell senescence and metabolic dysfunction is critical to understanding and treating age-related diseases. The propionate metabolism pathway is a highly conserved pathway critical for the metabolism of certain amino acids, odd chain fatty acids, cholesterol and propionate. The function of this pathway and its metabolites in cell senescence and aging remains to be identified. Previous studies and my preliminary data suggest that the activity of propionate metabolism changes with aging, and two metabolites in this pathway, methylmalonic acid (MMA) and propionyl-CoA (P-CoA), are involved in the regulation of cell senescence, lipid metabolism and protein modification. Therefore, I hypothesize that the propionate metabolism pathway plays a fundamental role in the aging process. During the K99 phase of this award, I will continue dissecting the role of MMA in cell senescence and aging-associated pulmonary fibrosis. Additionally, I will characterize a novel function of P-CoA in lipid droplet dynamics. I will also uncover the function of a novel post-translational modification (propionylation) on p53 (a key regulator for cell senescence) and catalog target proteins for propionylation. During the R00 phase of this award, I will expand on my lipid droplet studies and investigate the role or P-CoA in aging-associated ectopic fat accumulation (visceral fat deposition) and validate the target proteins for propionylation as identified during K99 phase. Furthermore, I will identify the main target genes and functions associated with histone H3 at lysine 23 (H3K23) propionylation during cell senescence. These investigations will pave the way for new research projects and serve as the basis for pursuing R01 grants and other funding opportunities. This award will allow me to expand on my training in aging physiology, metabolism and epigenetics, and gain expertise in chromatin biology, lipid droplets dynamic, and protein post-translational modification, ultimately helping me develop new niches (metabolic dysfunctions and aging) as an independent investigator. In addition to the scientific objectives outlined in this proposal, I have devised a comprehensive training plan for the K99 phase of the award to ensure a smooth transition towards independence.

NIH Research Projects · FY 2025 · 2024-07

Project Summary. We propose an innovative research education program, “Developing Research Leadership in Infectious Diseases Impacting Women.” Our goals are to address the knowledge gap in sex differences in global infectious diseases in the U.S and worldwide, and to train the next generation of clinical investigators in this field. In 2014, our team founded the Women’s Global Health Research Initiative which hosted two-day scientific conferences in global health and provided a high-impact training program for researchers in women’s health and global health. This initiative, now in its 9th year, has become a research community for both expert scientists and trainees in global health with over 80 faculty and 300 trainees. We now propose to leverage this initiative to expand educational training in infectious diseases that impact women and provide structured career development to a broader pool of early-stage investigators focused on global infectious diseases that impact women. Our program includes two components: (1) an annual “Global Infectious Diseases across a Woman’s Lifespan” course; and (2) a “Global Infectious Diseases and Women’s Health (GIDWH)” Scholars Program. The scientific course will provide didactic training in sex differences in global infectious diseases (HIV, TB, malaria, respiratory infections), host-immunology across the woman’s lifespan, treatment, prevention, and community factors that contribute to poor health outcomes for women in the U.S. and globally. The course will also provide practical tools to conduct research in women’s health, and networking opportunities with peers and senior leaders in the field. The GIDWH Scholars Program will consist of a competitively-selected group of early-stage investigators. Scholars will receive longitudinal training over two years to strengthen and complement ongoing research training with their primary scientific mentor at their home institution. GIDWH Scholar training will include peer mentorship cohorts for leadership training, and Research Advisory Teams of senior researchers with technical or topical expertise specific to the scholar’s research interests. Scholars will receive research-enabling funds to augment their career progression. Over five years, 200 early-stage investigators will attend the educational course, of whom 10 per year (years 1-4) will be selected to participate in the two-year GIDWH Scholars Program (40 total). Our proposed program aligns with the NIAID’s women’s health mission to “conduct and support basic and translational research to understand, diagnose, prevent and treat infectious diseases that impact the health of women and girls.” Through combining didactic research training with individualized leadership training and career development, we expect that trainees will improve their understanding of the complex biologic and other determinants that contribute to sex differences in infectious diseases. Further, we anticipate that this research education program will further their ability to conduct research to improve the diagnosis, prevention, and treatment of infectious diseases that impact women.

NIH Research Projects · FY 2024 · 2024-07

ABSTRACT The 2024 Annual Meeting of the Pan-American Society for Pigment Cell Research (PASPCR), entitled, “Where pigment cell research never sleeps” will be held in New York City, September 9-12, 2024. Founded in 1988 and made up of approximately 100 members, the PASPCR unites clinicians, developmental biologists, biochemists, immunologists, cell biologists, photobiologists and chemists in the study of pigmentation, melanin and melanocytes. The overall objective of PASPCR 2024 is to increase our knowledge of the normal and abnormal chemistry, biology, and photobiology of the melanocyte across species and its contribution to human health and disease. Building on the success of partnering with other melanocyte focused societies at our last meeting, we are planning a patient session sponsored by the Skin of Color Society (SOCS), the Global Vitiligo Foundation (GVF), and the Global Albinism Alliance (GAA) focused on patients with skin of color and the impact of melanocyte disease. The goals of the meeting are to (1) highlight recent advances in melanin structure and analysis across species, in pigment cell response to light and other environmental stimuli, and in genome-level understanding of melanocyte function and transformation using relevant animal models and human tissues, (2) bring together investigators and clinicians with strong interests in photoprotection, melanocyte developmental and stem cell biology and health disparities in pigmentary disease, and (3) provide mentorship to young investigators with a specific focus on identifying and overcoming career barriers encountered by women and underrepresented minorities. We have emphasized the importance of diversity, equity, and inclusivity (DEI) with a dedicated educational session and patient session, by inviting faculty from the SOCS to our meeting. In response to requests by trainees and young investigators, we are incorporating a panel discussion focused on pigment cell research in the Industry setting. We have ensured meaningful inclusion of young investigators, women, and underrepresented minorities among the invited speakers and leadership of PASPCR 2024. The meeting also features a "Mentorship Mixer" that provides a structured mechanism for students and trainees to obtain scientific and career guidance from well-established scientists. Led by a strong organizational team, PASPCR 2024 promises to be a conference of lasting impact for the pigment cell biology community.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Despite decades of research, the pathogenesis of Idiopathic Pulmonary Fibrosis (IPF) is still not completely understood. As a consequence, IPF causes significant morbidity and mortality and there are no therapies that reverse existing fibrosis. IPF is characterized by accumulation of collagen and fibrotic extracellular matrix in the lung that replace normal tissue and interfere with gas exchange, leading to dyspnea, respiratory failure, and death. A key knowledge gap preventing our development of effective therapeutics is the understanding of how collagen is degraded and turned over. If we were able to promote enhanced turnover and clearance of collagen in fibrotic IPF, this could provide a viable strategy to reverse fibrosis in IPF. This grant proposes to address this knowledge gap by delineating pathways of collagen clearance that we discovered by a recent unbiased CRISPR screening approach. We have found a previously unappreciated mechanism of regulation of collagen clearance: that collagen synthesis is sensed by cells internally and directly regulates clearance of collagen (i.e. cellular uptake and degradation of extracellular collagen). The sensing mechanism is dependent on ER resident protein SEL1L. This mechanism appears to be a homeostatic negative feedback loop to limit accumulation of collagen in tissues. Importantly, we have also found that the induction of collagen turnover by collagen synthesis is impaired in IPF, contributing to the excess and unmitigated buildup of collagen in lung tissue. The research proposed in this grant will fully define the upstream and downstream mechanisms governing this pathway as well as the cause of the impairment in this pathway in IPF. We will use mouse models, human lung organoids, human IPF tissue and cells, and in vitro and in vivo functional experiments. If successful, the proposed research has the potential to open the door to entirely new therapeutic avenues to try to reverse fibrosis in IPF and ameliorate symptoms in patients.

NIH Research Projects · FY 2025 · 2024-07

Abstract Transient receptor potential (TRP) channels are a large, eukaryotic ion channel superfamily that control diverse physiological functions, and are therefore attractive drug targets. To date, more than 210 structures from over 20 different TRP channels have been determined, all are tetramers. Despite the wealth of structural information there are many open questions, including the pore-dilation phenomenon, whereby prolonged activation leads to an increase in conductance, permeability to large ions, and loss of rectification. Using HS-AFM, we have discovered a hitherto unobserved pentameric state for TRPV3 that is in in a dynamic equilibrium with the tetrameric state through membrane diffusive exchange of protomers. Simple geometric considerations to estimate the pore size, as well as the similar timescale of seconds-to-minutes we observed necessary to achieve both the pentameric state and pore-dilation currents, suggest that the pentameric state may be the structural correlate to the so far elusive pore-dilation phenomenon. In this project we will therefore aim to (1) correlate the pentameric TRPV3 state with pore-dilation. (2) Determine an atomic structure of the pentameric state, and (3) investigate gain-of-function TRPV3 disease mutations which we hypothesize have increased occurrence of the pentameric state. To complete these aims we will: (1) Use HS-AFM and NanoDSF to assess whether there is an increase in the population of pentamers, and a decrease in the stability the tetramers, following the addition of different well-known pore-dilation agents (diphenylboronic anhydride (DPBA) and heat (45°C)), and other pore-dilation agents that we will discover. (2) Use molecular dynamics simulations (MDS) for equilibration of an initial pentameric model and determine the experimental pentameric structure using cryo-EM to achieve a high- resolution understanding of the pore structure and protomer interfaces in the pentamer. (3) Study two gain-of- function mutations (M572I and Q580P) associated with the Olmsted syndrome, located at the TRPV3 protomer interface, and use HS-AFM, cryo-EM and electrophysiology measurements to assess whether these mutations increase the likelihood for the pentameric and pore-dilated state. Completion of these aims will allow us to assess whether the increased conductance, permeability to larger ions, and loss of rectification associated with pore dilation are indeed related to a change in the oligomeric state of TRP channels, and thus provide a correlate to the yet structurally undetermined phenomenon of pore-dilation. Additionally, this work will provide a structural explanation and mechanism for the increased channel activity following different TRPV3-related mutations and will thus further the development of drugs and therapeutic tools for treatment of TRP-channel related diseases. The results from this project are expected to provide answers to some long-standing and unresolved questions in the field of TRP-channel research, and to pave the way for new avenues in membrane protein research in general, considering membrane diffusive protomer exchange as a process for conformation variability.

NIH Research Projects · FY 2025 · 2024-07

Project summary Currently, there is no cure for intrahepatic bile duct (IHBD) paucity except for liver transplantation. IHBD paucity causes accumulation of pathologic bile within the liver resulting in bile stasis and over time can lead to chronic cholestasis. Diseases such as Alagille syndrome (ALGS), biliary atresia, progressive familial intrahepatic cholestasis, alpha1-antitrypsin deficiency, primary biliary cirrhosis, and primary sclerosing cholangitis are associated with IHBD paucity. The standard of care for some of these diseases includes reducing bile acids through surgical biliary diversion, inhibition of the ileal bile acid transporter (i.e., IBAT) or by altering the composition of the bile acid pool (using bile acids salts or FXR ligand agonists) to interrupt and modify enterohepatic circulation. Currently, there are no clinical approaches to augment IHBD architecture or bile duct number in patients with these diseases. Therefore, in the absence of new approaches, patients with IHBD paucity will remain dependent on liver transplantation as definitive therapy for the foreseeable future. We have published that hepatocyte-to-cholangiocyte transdifferentiation in the absence of epithelial Notch activity is competent through a Tgfbr2-dependent mechanism. However, hepatocyte-to-cholangiocyte transdifferentiation is inefficient in patients with ALGS, due to global JAGGED1 (JAG1) haploinsufficiency with a significant number of patients requiring liver transplantation. Recent lineage tracing and single-cell RNA sequencing studies inform us that even though arising from a common mesenchymal progenitor, the identity of periportal mesenchyme is divergent from hepatic stellate cells and a critical knowledge gap in all aspects of IHBD development. Preliminary work strongly supports a role for periportal mesenchyme in this process, but the underlying role of periportal mesenchyme in IHBD paucity is superficial at best, especially regarding the mechanisms regulating biliary epithelial-periportal mesenchymal crosstalk. Our long-term goal is to understand the mechanisms regulating epithelial-mesenchymal crosstalk during IHBD specification, morphogenesis, maintenance, and response to injury/disease. Based on published and unpublished results we propose to address the central hypothesis that Jag1 haploinsufficiency impacts the periportal mesenchyme indirectly by reducing Notch activity in the hepatic epithelium hindering the implementation of the full cholangiocyte transcriptional program and thereby influencing the periportal mesenchyme by disrupting optimum crosstalk between the epithelium and mesenchyme. Our aims are to: 1) define the role Gli1 periportal mesenchyme plays to regulate epithelial IHBD development, and 2) define the role epithelial Ihh plays to regulate epithelial-periportal mesenchymal crosstalk. Long-term, the improved understanding of epithelial-periportal mesenchymal crosstalk will enable the development of targeted therapies capable of altering and augmenting IHBD disease.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT The goal of this project is to evaluate the temporal and predictive association of emotion reactivity (ER) and cognitive reappraisal (CR) (a well-studied emotion regulation strategy) with intensity and duration of suicidal ideation (SI) in middle-aged and older adults following their discharge from a suicide-related hospitalization, a period of high suicide risk. Our preliminary Event Related Potential (ERP) and self-reported affect data on middle-aged : a) increased ER and reduced CR at Discharge were independently associated with increased intensity and longer duration of suicidal ideation at Week 6; and b) patients with combined high ER and low CR at Discharge had higher intensity and longer duration of suicidal ideation at Week 6 than the rest of the patients. and older adults (N=36) who were hospitalized for suicidality demonstrated that We will recruit 162 middle-aged and older adults (40-90 years old) who have been hospitalized for suicidal ideation or suicidal behavior [Columbia Suicide Severity Rating Scale (C-SSRS) at Admission > 1, i.e., 1=Wishes to be Dead]. We will follow the participants for 6 months after discharge. Suicidal ideation and behavior will be measured weekly through an interviewer-administered instrument (C-SSRS). ER and CR will be assessed with: 1) electrocortical measures (Late Positive Potential) and self-reported affect during an emotion reactivity and regulation EEG task (primary measures), and 2) the Emotion Regulation Questionnaire and the Perth Emotional Reactivity Scale (secondary measures). Assessments will be conducted at Admission (without EEG), Discharge, and at Weeks 6, 12, 18, and 24. 1. Examine the Independent Effects of ER and CR on Intensity and Duration of SI. 2. Examine the Combined Effects of ER and CR on Intensity and Duration of SI. Exploratory Analyses: 1. Examine whether demographic variables, discharge clinical characteristics, rehospitalization, medical comorbidities, and other suicide risk factors moderate the relationship of ER and CR with suicidal ideation. 2. Examine association of the longitudinal profile of ER and CR with the longitudinal profile of intensity and duration of suicidal ideation. Primary Aims: This is the first study to examine the predictive relationship of ER and CR with suicidal ideation in middle-aged and older adults at high suicide risk. Understanding this relationship will help identify: a) mechanisms which increase suicidal ideation, and b) subgroups of patients at increased suicide risk. Consistent with the concept of “Precision Medicine”, the findings may contribute to the development of personalized psychosocial interventions for this population.

NIH Research Projects · FY 2026 · 2024-07

ABSTRACT The goal of this project is to evaluate the temporal and predictive association of emotion reactivity (ER) and cognitive reappraisal (CR) (a well-studied emotion regulation strategy) with intensity and duration of suicidal ideation (SI) in middle-aged and older adults following their discharge from a suicide-related hospitalization, a period of high suicide risk. Our preliminary Event Related Potential (ERP) and self-reported affect data on middle-aged : a) increased ER and reduced CR at Discharge were independently associated with increased intensity and longer duration of suicidal ideation at Week 6; and b) patients with combined high ER and low CR at Discharge had higher intensity and longer duration of suicidal ideation at Week 6 than the rest of the patients. and older adults (N=36) who were hospitalized for suicidality demonstrated that We will recruit 162 middle-aged and older adults (40-90 years old) who have been hospitalized for suicidal ideation or suicidal behavior [Columbia Suicide Severity Rating Scale (C-SSRS) at Admission > 1, i.e., 1=Wishes to be Dead]. We will follow the participants for 6 months after discharge. Suicidal ideation and behavior will be measured weekly through an interviewer-administered instrument (C-SSRS). ER and CR will be assessed with: 1) electrocortical measures (Late Positive Potential) and self-reported affect during an emotion reactivity and regulation EEG task (primary measures), and 2) the Emotion Regulation Questionnaire and the Perth Emotional Reactivity Scale (secondary measures). Assessments will be conducted at Admission (without EEG), Discharge, and at Weeks 6, 12, 18, and 24. 1. Examine the Independent Effects of ER and CR on Intensity and Duration of SI. 2. Examine the Combined Effects of ER and CR on Intensity and Duration of SI. Exploratory Analyses: 1. Examine whether demographic variables, discharge clinical characteristics, rehospitalization, medical comorbidities, and other suicide risk factors moderate the relationship of ER and CR with suicidal ideation. 2. Examine association of the longitudinal profile of ER and CR with the longitudinal profile of intensity and duration of suicidal ideation. Primary Aims: This is the first study to examine the predictive relationship of ER and CR with suicidal ideation in middle-aged and older adults at high suicide risk. Understanding this relationship will help identify: a) mechanisms which increase suicidal ideation, and b) subgroups of patients at increased suicide risk. Consistent with the concept of “Precision Medicine”, the findings may contribute to the development of personalized psychosocial interventions for this population.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Research: Inflammatory bowel diseases (IBD) are a group of chronic inflammatory diseases of the gastrointestinal tract that affect over 3 million Americans. Abdominal pain is a common symptom of IBD, but the role of pain-sensing nociceptor neurons in regulating the inflammatory process is largely unknown. Based on my published and new preliminary data, I hypothesize that nociceptor sensory neurons and neuropeptides are tissue-protective, but become dysregulated in the context IBD, and subsequently drive pathological adaptive immunity and alterations to the microbiota that facilitate intestinal inflammation and impair tissue repair. This proposal aims to thoroughly investigate the regulation of intestinal infection and inflammation by nociceptor sensory neurons and determine the detailed underlying cellular and molecular mechanisms using innovative and interdisciplinary approaches from neurobiology, gastroenterology, immunology and microbiology. These studies will uncover novel neuro-immune-microbiota crosstalk pathways and their unique roles in intestinal inflammation and transform our understandings of host-microbe interactions and neuro-immune mechanisms in IBD. Career goals: My overarching career goal is to become an independent investigator at an academic institution where I decipher the neuro-immune and neuron-microbiota pathways that regulate intestinal tolerance and inflammation and eventually discover neuronal-based and microbiota-based drug targets for immune modulation to treat chronic inflammatory diseases including IBD. Furthermore, I aspire to become an inspirational teacher and outstanding mentor. Career Development Plan: To become a successful independent investigator I will further develop various research, professional and personal skills. These will include further acquiring expertise in neurobiology and microbiology techniques, as well as fostering collaborations and developing skills in writing, mentoring, teaching, communicating and laboratory management. My host laboratory and Weill Cornell Medicine provides an outstanding academic environment in which trainees can fulfil these criteria by promoting scientific interactions and enrolling in courses to develop skills in specialist scientific areas, personal development and laboratory management. The mentorship I will receive will be of the highest standard. Furthermore, in addition to my mentor and co-mentor, I have support from a team of independent investigators with extensive expertise in areas that will greatly facilitate the completion of experiments proposed and my career transitional into independence. Career Development Environment: I will perform the K99 phase in the laboratory of Dr. David Artis at the Weill Cornell Medicine, which provides an unprecedented environment in terms of the resources and facilities available. Weill Cornell Medicine is an ideal environment to develop during the K99 phase as I progress to become a successful independent investigator. The laboratory has access to all of the equipment and facilities necessary to complete the experiments proposed in this application.

NIH Research Projects · FY 2025 · 2024-07

Introduction: The overall goal of our research project is to reduce the mortality and morbidity of children with acute and emergency illnesses by improving care during ambulance transportation using telemedicine. In the R21 phase, we proved the feasibility of conducting an ambulance-based telemedicine cluster randomized trial. Specific Aims for the R33 phase: The R21 phase had three aims, while the R33 phase have the following two aims, which remain unchanged from our initial proposal. - SA 4: To test the efficacy of Ambulance Based Teleconsultation (ABT) by measuring a change in the Pediatric Early Warning Signs (PEWS) of acutely ill children from the scene of injury/illness to the pediatric emergency department (PED) through a cluster-randomized trial. - SA 5: To develop the capacity of local partners in m-health research capacity. Methodology: SA4 will be a prospective cluster randomized trial with each ambulance serving as a cluster. We will use a stratified random sampling design to assign 30 ambulances to receive the ABT setup, while 30 others will serve as control clusters. Our primary outcome will be a change in the Pediatric Early Warning Score from the scene of illness/injury to the hospital triage/resuscitation room. Our secondary outcomes will be % of completed calls, the satisfaction rate of EMTs and Telemedicine Physicians, and the outcome at the end of the ED visit. We will work with our existing partners and use the same telemedicine setup as tested in the R21 phase. We will use the SIEHS EMTs who were part of the intervention and control ambulances during the R21 phase to work with the research team as trainers for the rest of the service. Our minimum required sample size of 600 (300 each in intervention and control clusters) remains unchanged. The only change in the data collection process will be the incorporation of Emergency Department PEWS in the standard electronic medical record by the ChildLife Foundation. This will ensure consistent and timely access to PEWS scores during the trial. As tested in the R21 phase, we will store identifiable data in a codebook on a password-protected institutional drive accessible to the research team only. After both sets of PEWS have been completed and follow-up consent has been received, the data will be de-identified and made accessible for analysis. Concurrently, we will share data on outcomes with the currently existing Data Safety Monitoring Board at 10%, 25%, 50%, 75%, and 100% levels of enrollment. Table 1 summarizes the milestones for SA4. For SA5, we will build capacity on telemedicine/telehealth research and will further strengthen expertise in emergency care clinical trials in Pakistan. This will be achieved through quarterly webinars, two workshops, and one national seminar. Additionally, we plan to support the training of SIEHS ambulance in modern prehospital care systems through one study visit to a regional or international center of excellence. Expected Outcomes: The findings from this research are expected to yield critical safety and efficacy data, which will serve to address the shortage of well-trained medical personnel in pediatric emergency care within low- and middle-income countries, potentially impacting more than 50% of global pediatric mortality. Furthermore, the insights gained from this study can have significant relevance for underserved rural communities in high-income countries, such as the United States. Particularly in regions where ambulance services heavily rely on the commitment of part-time community volunteers, often operating across extended distances.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT: Glioblastoma (GBM), the most prevalent malignant primary brain tumor, is an extremely aggressive form of diffuse glioma originating from astrocytic lineage. Despite recent advancements in multimodal GBM therapy, which incorporates surgery, radiotherapy, chemotherapy, and supportive care, the overall prognosis remains dismal, and long-term survival is rare. Immunotherapy holds promise in leveraging the immune system to target and eliminate brain tumor cells. However, the highly immunosuppressive environment within GBM represents a critical impediment to successful immunotherapy. SLIT2/ROBO signaling is a novel immune evasion mechanism in the tumor microenvironment of GBM. High SLIT2 expression in GBM patients results in the accumulation of immunosuppressive tumor-associated macrophages (TAMs) as well as vascular dysmorphia. This is further supported by the prevention of TAM tumor-supportive polarization and angiogenic gene expression upon systemic SLIT2 inhibition, resulting in improved tumor vessel function and enhanced efficacy of chemotherapy and immunotherapy in GBM mouse models. Therapeutic targeting of SLIT2/ROBO interaction is currently restricted to biologics, and there are no active clinical trials for GBM evaluating SLIT2/ROBO inhibition as a therapeutic strategy. In comparison to biologics, small molecules will minimize the immunogenicity risk, enable better management of adverse events (AEs) based on their amenability for pharmacokinetic optimization, and hold promise for central nervous system (CNS) penetration. In response to PAR-23-264, our three-year proposal aims to establish a novel macrophage-based immunotherapy approach for GBM based on targeting SLIT2/ROBO interaction with small molecules, which may synergize with current FDA-approved therapies for GBM. Our expertise in assay development, high-throughput screening (HTS), discovery of small molecule immunomodulators, hit-to-lead optimization, and immunopharmacology uniquely positions us to achieve this goal. Aim 1 will complete the screening of a CNS-focused chemical library of small molecules for SLIT2 binding, followed by an evaluation of the ability of the hits to inhibit SLIT2/ROBO interaction using fluorescence-based assays. Aim 2A will validate the top hit compounds as SLIT2/ROBO inhibitors using a panel of cell-free and cell-based assays, including an in vitro spheroid invasion assay using patient-derived cells from GBM tissue. Aim 2B will perform exploratory medicinal chemistry and preliminary structural optimization of the top validated hits in order to guide future extensive optimization. The proximal expected outcome of this work is introducing first-in-class small molecule SLIT2/ROBO inhibitors as candidates for preclinical evaluation, particularly within innovative combination therapies for GBM.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Genomic instability, referring to a heightened frequency of mutations and DNA damage, is a hallmark of cancer. A major consequence of cancer-intrinsic genomic instability is the activation of innate immune response, which can confer an antitumor state to cells. However, many cancers co-opt regulatory pathways to escape immune surveillance. Our long-term goal is to elucidate the molecular mechanisms of how innate immune consequences of genomic instability are regulated. Particularly, our group studies the immunogenic potential of cytosolic DNA, a molecular signal that arises from mitotic and nuclear aberrations. Cytosolic DNA activates a DNA sensing pathway termed cGAS-STING to trigger the type I interferon response. Three prime repair exonuclease 1 (TREX1) suppresses this pathway by degrading cytosolic DNA. Indeed, many cancers upregulate TREX1 to accommodate cytosolic DNA without immune activation. Yet, TREX1-null mutations are linked to inflammatory disorders like Aicardi-Goutières syndrome (AGS). Further, frameshift mutations that cause ectopic TREX1 nuclear localization are linked to retinal vasculopathy with cerebral leukodystrophy (RVCL). Therefore, TREX1 plays a complex role in human diseases through degradation of cytosolic DNA, but molecular mechanisms of TREX1 function are poorly understood. The goals of my project are to characterize the TREX1- specific elements, the polyproline helix (PPII) and the C-terminal intrinsically disordered region (C-IDR), in the context of immune regulation (Aim 1); and to dissect the impacts of ectopic TREX1 localization in the nucleus (Aim 2). My working hypothesis is that these TREX1-specific non-catalytic domains are essential for recruiting protein partners that prime DNA substrates for digestion by TREX1, thus enhancing cGAS-STING regulation. Further, I expect that PPII induces de novo interactions when TREX1 is ectopically expressed in the nucleus. I propose to test my working hypothesis using molecular and cellular approaches in human cells to quantify the activation of cGAS-STING, to identify protein-interacting domains of TREX1, and to examine the behavior of nuclear TREX1. This project will characterize novel TREX1 structural features that confer its unique immunosuppressive role, as well as explain how ectopic TREX1 localization can induces disease. It will also identify novel TREX1 interactors, contributing to the discovery of molecular targets in cancer therapy.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Early life exposure to microbes is necessary for normal immune development, and clinical studies link alterations in early life bacterial composition to subsequent development of inflammatory disorders such as inflammatory bowel disease (IBD) and atopy such as asthma and dermatitis. In intestinal disease, microbiota- specific T cells can drive tissue pathology. Therefore, it is crucial to understand the homeostatic mechanisms that establish immune tolerance to commensal microbes during infancy in order to address inflammation in adulthood. As antigen presenting cells (APCs) initiate the T-cell response, we want to elucidate how they respond to microbes during early life. This work will have broad implications for development of therapies that address the root of inflammatory disease including reshaping of pathogenic T cell responses. The proposed project aims to describe how microbial colonization during early life drives the phenotype and function of mononuclear phagocytes (MNP), which are a lineage of APCs known to coordinate host-microbe responses. This lineage consists of monocytes that circulate in the blood, and the dendritic cells (DC) and macrophages that they differentiate into upon entering tissue. While the factors that dictate this fate decision remain undefined, these diverging cell fates lead to functionally different outcomes. DCs migrate to lymph nodes to initiate T-cell responses while macrophages remain resident in the tissue to clear microbes and support barrier function. Data from our lab shows that in early life but not adulthood, intestinal DCs expand and bring microbial antigens to the thymus where they induce proliferation of microbe-specific T-cells. Interestingly in the early life mesenteric lymph node, we see similar DC expansion and microbial trafficking but do not see the corresponding T-cell response. This data led to our hypothesis that during early life, mononuclear phagocyte differentiation is skewed towards a DC fate. We further hypothesize that early life lymphoid organs provide signals to instruct microbial antigen processing and presentation resulting in life span and organ restriction of expansion of microbiota recognizing T cells. To address our hypothesis, in Aim 1 we will utilize single-cell RNA sequencing and adoptive monocyte transfer to understand cell intrinsic and tissue regulation of monocyte fate over the lifespan. In Aim 2, by using functional readouts, we will interrogate changes in DC function over the lifespan. Together, our work will reveal how MNPs mediate T-cell responses to microbes during development, further advancing our ability to address inflammation during adulthood.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Vertebrae and long bones display marked differences across a range of clinical and basic properties, such as their basic architecture, responsiveness to PTH, their rate of seeding with solid tumor metastases, or the types of developmental and degenerative diseases present. The etiology of these differences has remained elusive, in part due to the current state of skeletal stem cell (SSC) biology that largely considers skeletal stem cells in a monolithic manner, without drawing clear distinctions in cell type based on bone of origin. Here, we have identified a vertebral skeletal stem cell (vSSC), having established that it displays a comprehensive set of stemness features, including the ability to self-renew across multiple rounds of transplantation, the ability to serially form bone organoids across multiple rounds of transplantation and reisolation, long-term label retention after a pulse with a chromatin bound H2B-GFP marker, in vivo multipotency through an ability to give rise to chondrocytes, osteoblasts and adipocyte in vivo, and being long-lived in vivo. This vSSC type is specific for vertebrae, being completely absent from long bones. These vSSCs have a clear physiologic contribution to vertebral mineralization, as deleting genes required for osteoblast differentiation with a Zic1- cre created for this project targeting the lineage of this vSSC produced marked defects in the mineralization of both the dorsal neural arch and the vertebral body and no detectable phenotype in long bones. Thus, we have here identified a vSSC, including generating 2 cre lines targeting this cell, that meets all published criteria to be defined as a novel SSC type. Here, we propose to address a series of key issues enabled by this discovery to further develop the clinical and physiologic role of this new stem cell type. First, (Aim 1) we will determine the unique cellular features of the vertebral bone versus long bones, demonstrate which of these features are due to Zic1+vSSCs, the relationship of Zic1+vSSCs to other skeletal progenitor cell types identified in the literature and the function of vSSC-specific transcription factors and secreted ligands. Next (Aim 2), we will establish that this vSSC is a key cell mediating spine fusion, a common orthopedic management strategy to treat a variety of spine degenerative disorders. Lastly (Aim 3), we will build upon preliminary efforts identifying a human counterpart of the murine vSSC to fully establish the identity and differentiation hierarchy of the human vSSC and to determine the functional conservation of vSSC-lineage defining transcription factors. Altogether, this project will establish the existence and unique properties of a new and distinct vSSC cell type in humans and mice and facilitate subsequent studies into how this vSSC mediates vertebral skeletal disorders or how these disorders can be therapeutically addressed by targeting this new stem cell.

NIH Research Projects · FY 2025 · 2024-07

The mission of the Tri-Institutional MD-PhD Program (Tri-I) is to train a talented group of clinician-scientists prepared to bridge the gap between laboratory research and clinical medicine. Graduates of the program are well-grounded in human biology, pathophysiology, and clinical medicine and are endowed with an advanced understanding of biomedical science, as well as a mastery of critical thinking and experimental skills. These skills will allow them to undertake complex, interdisciplinary, quantitative, and collaborative studies to elucidate basic biological processes pertaining to human health and disease and to transfer advances in research to the understanding, prevention, and treatment of human disease. Tri-I is a joint undertaking between Weill Cornell Medicine (WCM), The Rockefeller University (RU), and Memorial Sloan Kettering Cancer Center (MSK). Trainees complete their MD degree at Weill Cornell Medical College (WCMC), and PhD training at one of the three participating graduate schools: Weill Cornell Graduate School of Medical Sciences (WCGS); the David Rockefeller Graduate Program in Bioscience at RU; or the Louis V. Gerstner Jr. Graduate School of Biomedical Sciences (GSK) at MSK. An integrated curriculum reinforces the students’ identities as clinician-scientists. In the first two years in the program, MD-PhD-specific graduate-level coursework, including training in quantitative methods, responsible conduct of research, and rigor and reproducibility, is integrated with pre-clinical medical school courses and 18 weeks of core clinical clerkships. Students complete three laboratory research rotations in at least two different institutions before selecting a thesis lab and enrolling in the graduate school at which their thesis mentor has their primary appointment. PhD requirements are comparable for all MD-PhD students across graduate institutions, and trainees may take courses for credit in any of the graduate schools. After they have defended their thesis, trainees complete their remaining clinical training at WCMC. Over 200 participating faculty members serve as mentors and undergo training to ensure a safe training environment, responsible conduct of research, and scientific rigor and reproducibility. Students receive multi-layered mentorship and advising, particularly around transitions to and from the laboratory and in preparation for post-graduate residency training. Based on Tri-I’s high trainee retention in the program and in academia and biomedical research following graduation, this application requests funds to support 42 trainees per year. No trainee will be appointed to the grant for more than a total of four years. The student body averages 150 MD-PhD trainees. Over the past 50 years, over 500 MD-PhDs have graduated from Tri-I and its predecessors. In the past 15 years, 85% of graduates have appointments in medical schools, research institutes, or biotech/pharma. Tri-I is committed to maintaining its longstanding tradition of training future leaders of biomedical research.

NIH Research Projects · FY 2026 · 2024-07

Project Summary / Abstract Biological systems operate within fluctuating environments, and, therefore, are inherently tasked with accurately responding in real-time to a myriad of signals. Recent evidence suggests that a memory of inflammation can be encoded and retained in the epigenome of cells even following resolution of the initial stimulus. The presence of `inflammatory memory' suggests that preservation of tissue homeostasis also incorporates an evolutionary adaptation in which future responses are educated by past experiences. Inflammation is essential to the disruption of tissue homeostasis, and, in the pancreas, can destabilize the identity of terminally differentiated acinar cells. Recently, we have employed lineage-traced mouse models to delineate the chromatin dynamics that accompany the cycle of metaplasia and regeneration following pancreatitis, and unveiled the presence of an epigenetic memory of inflammation in the pancreatic acinar cell compartment. We have observed that despite histologic resolution of pancreatitis, acinar cells fail to return to their molecular baseline after several months, representing an incomplete cell fate decision – one wherein there is persistent MAPK signaling, AP-1 activation, and IL-33 in the pancreas microenvironment. In vivo, this epigenetic memory controls lineage plasticity, with diminished metaplasia in response to a second inflammatory insult but increased tumorigenesis with an oncogenic Kras mutation. We have demonstrated that both persistent chromatin and transcriptional changes constituting memory can be specifically recalled in the response to oncogenic stress. Together, our findings have defined the dynamics and recall of an epigenetic memory of inflammation that impacts cell fate decisions. In this proposal, we focus on building an understanding of the molecular underpinnings of memory of inflammation. Specifically, we will leverage our lineage-traced mouse models of pancreatic inflammation to establish an understanding of AP-1 binding to chromatin and its interaction partners (Aim 1). We will also employ specific genetic and pharmacologic perturbations to AP-1 factors and MAPK signaling in vivo to articulate whether epigenetic memory is reversible (i.e. inducing epigenetic `amnesia') (Aim 2). Third, we will examine the role of IL-33 as a cell-extrinsic factor driving the memory phenotype (Aim 3). Together our studies will define the molecular mechanisms that govern epigenetic memory of inflammation in the pancreas. In so doing, we expect to uncover the key cell types, transcription factors, and signaling intermediates that lead to persistent molecular alterations following transient injury. In turn, this work will shed new light in to how memory can be targeted to abrogate the diminished threshold for tumorigenesis. By analyzing the rational means for inducing epigenetic `amnesia', we will exploit the therapeutic opportunity that the durability of epigenetic memory offers to address the lasting implications of an inflammatory insult.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Research: Cancer is the second leading cause of mortality in the world, with estimates as high as 1.9 million new diagnoses and 0.6 million deaths in the United States last year. Trillions of normally beneficial microbes, termed the microbiota, continuously colonize the mammalian intestine and emerging evidence indicates key roles for these microbes in impacting cancer. This includes both promoting tumor growth and progression, but also in supporting response to immunotherapies, raising the exciting possibility to manipulate microbiota to combat cancer, however the mechanisms by which microbiota influence cancer progression and immunotherapies remain poorly understood. Based on my published and new preliminary data, I hypothesize that host-microbiota interactions (i) promote tumor growth by limiting metabolites that provoke anti-tumor immunity, and (ii) shape systemic anti-tumor immunity and the outcome of immunotherapy by antigen-specific responses to gut microbiota via crosstalk of RORgt+ lymphocytes. These studies will define two novel pathways by which microbiota interacts with mammalian hosts to impact tumor growth, progression, and immunotherapy response, thus provoking novel opportunities for intervention. Career goals: My overarching goal is to become an independent investigator at an academic institution where I investigate the mechanisms by which host-microbe interactions modulate anti-tumor immunity and immunotherapy response for the treatment of cancer. Further, I aspire to become an inspirational and productive mentor as these are essential aspects of being an independent investigator. Career Development Plan: To become a successful independent investigator, I will further develop a various research, professional, and personal skills. These will include further acquiring expertise in cancer biology and microbiology techniques, as well as fostering collaborations and developing skills in writing, communicating, teaching, mentoring and laboratory management. My host laboratory and Weill Cornell Medicine provide an outstanding academic environment where trainees can fulfil these criteria by fostering scientific collaboration and enrolling in courses to develop skills in specialist scientific areas, personal development, and laboratory management. The mentorship I will receive will be of the highest standard. Further, in addition to my mentor and co-mentor, I have support from a team of independent investigators with extensive expertise in areas that will greatly facilitate the completion of experiments proposed and my career transition into independence. Career Development Environment: I will perform the K99 phase in the laboratory of Dr. Gregory Sonnenberg at Weill Cornell Medicine, which provides an outstanding environment in terms of the resources and facilities available. Weill Cornell Medicine is an ideal environment to develop during the K99 phase as I progress to become a successful independent investigator. The laboratory has access to all the instruments and facilities necessary to complete the experiments proposed in this application.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Despite advances in the development of highly effective androgen receptor (AR)-directed therapies for the treatment of men with advanced prostate cancer, acquired resistance ultimately ensues. Lineage plasticity has been proposed as one mechanism of therapeutic resistance whereby patients with resistant disease develop AR-negative, androgen signaling-indifferent prostate tumors that lose their luminal identity and display neuroendocrine features (neuroendocrine prostate cancer, NEPC). While NEPC tumors share many genetic alterations with prostate adenocarcinoma, the potential drivers of lineage plasticity remain understudied. Using a novel genetically-engineered mouse model that faithfully recapitulates the transition to NEPC, I have established an organoid-based allograft platform that is amenable to gene editing technologies. Using single-cell based approaches, Furthermore, I have identified a previously undescribed tumor subpopulation with a unique transcriptional regulator that may represent an transition between adenocarcinoma and NEPC. For the proposed studies, I will modulate the expression levels of the identified transcriptional regulator using CRISPR-based gene editing or overexpression strategies in prostate organoids and determine how the transition to NEPC is affected. I will also assess the sensitivity of this tumor subpopulation to clinically-relevant treatment options, including androgen withdrawal and AR-targeted therapy. Finally, I will reveal how targeting epigenetic modifiers changes the composition of tumor subpopulations and reverses the development of therapeutic resistance. Alongside these scientific aims, I will use the period of support to enhance my skillset and develop as an independent researcher. Through a comprehensive plan, including workshops, course work, clinical case conferences, and attendance at seminars and scientific conferences, I plan to develop a deeper understanding of bioinformatics at the single cell level, expand my exposure to critical barriers facing clinicians and prostate cancer patients, and successfully continue my transition to an independent research position. The environment at Weill Cornell Medicine, and among its closely aligned neighboring institutions, is ideal for me to complete the proposed studies and will help foster my continued research and career development success.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Colorectal cancer (CRC) is a devastating disease with no effective therapeutic options for patients suffering from advanced metastasis. Immune checkpoint inhibitors (ICIs) offer promising treatment potential for a small subset of patients with microsatellite instable (MSI)/mismatch repair deficient (dMMR) tumors. However, most CRC patients have microsatellite stable (MSS)/mismatch repair proficient (pMMR) tumors that respond poorly to ICIs. There is therefore an urgent need to identify novel targets and biomarkers of response to immunotherapy in MSS tumors, which constitute over 90% of metastatic CRC. ZFP36L2, an RNA-binding protein mutated in 5-10% of CRC, is a member of the TIS11 gene family which is known to suppress inflammation by blocking the expression of proinflammatory chemokine and cytokine mRNAs in immune cells. Preliminary data from our lab suggests that loss of ZFP36L2 function in human CRC leads to increased T cell recruitment into the normally immunologically “cold” tumor microenvironment (TME) of MSS/dMMR TME. These data suggest that ZFP36L2 may also suppress the translation of cytokines and chemokines in CRC as a mechanism of immune evasion, and further that ZFP36L2 mutations may give rise to increased immune cell infiltration into CRC tumors, which may enhance responses to immunotherapy. To address this, we have developed a novel syngeneic mouse ZFP36L2 knockout CRC organoid model and a ZFP36L2 knockout genetically engineered mouse CRC model. These tools will allow us to delineate the ability of ZFP36L2 to regulate the inflammatory secretome of CRC (Aim1) and assess how ZFP36L2 mutations impact the composition and functionality of the TME in CRC (Aim 2). This research will elucidate the functional consequences of mutations in ZFP36L2, whose function in human cancer is poorly understood. More broadly, our work will increase our understanding of how tumors shape their local immune environment to support disease progression. Our interrogation of the role of ZFP36L2 in CRC will address critical unmet needs to develop novel treatment strategies for MSS CRC and identify clinically actionable biomarkers of response to immunotherapy. This work has the potential to significantly improve the survival and quality of life for CRC patients.

NIH Research Projects · FY 2024 · 2024-06

PROJECT SUMMARY Glucose transporter 1 deficiency (G1D) constitutes an increasingly recognized cause of epilepsy, intellectual disability and motor control dysfunction. This summit proposal will catalyze the conjunction of several important but still disjointed developments in need of cross-fertilization. Rapid scientific, patient organization and information dissemination advances have made, for the first time, research on G1D approachable from a meaningful, patient-centered perspective. This means that harmonizing the collective voices of the scientific and patient community represented by the Glut1 Deficiency Foundation can greatly propel further progress. Although prevalence estimates are likely to change, about 1% of idiopathic generalized epilepsy or 1 in 23,000 births have been associated with G1D. Nevertheless, there is still no effective treatment. Yet, our recently adopted Team Science perspective and work in numerous single laboratories elsewhere have been yielding steady benefits. For example, we have contributed from the molecular characterization of mutation pathogenicity to the isolation of a key neural circuit involved in epileptogenesis, including the first rigorous trial of a dietary treatment, all under NIH sponsorship. Still, numerous additional approaches worldwide have also proven as fruitful or promising. Therefore, we propose to join the patient community with many of the leading investigators in the field in a summit. An example of the effectiveness of such collaborative work are our 2020 international diagnostic and treatment consensus recommendations, which already stand in need of revision given how rapidly the field is advancing. We also aim to reach consensus, or at least lay the groundwork, on the best animal models to share, the most informative biomarkers, the main disease phenotypes and their evolution across the lifespan, and the most effective clinical trial and data sharing approaches, given the need to unify concepts and disseminate resources. None of this can be accomplished by any single laboratory or team working in isolation. Instead, our approach will be open, transparent and international. The summit will benefit from 1) extensive previous expertise with scientific and patient centered G1D conferences and additional multi-lingual online events organized according to rigorous metrics, 2) existing partnerships with sister G1D patient organizations across the world, 3) the accessibility of the proposed location, which has been home to the largest clinic focused on G1D, 4) the strong support of some of the leading researchers not only in G1D but also in related fields such as dementia, cancer and epilepsy where metabolism is important, and 5) robust diversity and event safety plans. We expect ample dissemination of the proceedings via several mechanisms, thereby propelling a transformation of the G1D field for years to come, including the development of a network of investigators informed by patient needs and the creation of a framework for future revisions.

NIH Research Projects · FY 2025 · 2024-06

Project Summary Clonal expansions increase with age throughout many tissues in the body. These outgrowths are driven by somatic mutations that provide a selective advantage over wild-type cells within a specific microenvironment. Clonal expansions radically increase with age, and are also shaped by disease-related factors. How these factors and clonal mosaicism (CM), reciprocally influences each other constitute some of the most important biological questions in current human genetics. A detailed understanding of the biology of aging at the cellular and molecular level is needed for the development of potential mitigation therapies that target aging morbidities. To make progress toward this goal, we need enhanced ability to identify and characterize recurrent somatic mutations in both healthy and diseased aging tissues at high-resolution. As identification of clones is hampered by low tissue availability and resource-intensive DNA sequencing methods, conceptual and technical innovation overcoming these challenges could rapidly accelerate the pace of variant discovery. Once discovered, functional characterization of these clonal variants in genetically heterogenous tissues requires application of genotype- aware, multi-modality methods in primary human samples to link genotypes and epigenetic profiles of individual cells. Currently, no such tools exist to characterize multiple molecular features of the somatic epigenome. To address these important gaps, we aim to (i) develop tools for the identification of candidate somatic variants at scale from publicly available data and (ii) apply novel technologies that map genotype to epigenetic phenotype in single cells, enabling analysis of multiple tissues from clinical samples obtained from aging individuals. Specifically, we aim to harness advancements in machine learning to develop a tool that can accurately identify somatic variants in bulk RNA-seq and apply it to publicly available data in healthy and diseased tissue from aged individuals. We believe that we can gain valuable information about the clonal architecture of diabetes, obesity, hypertension and other age-related diseases from our public databases at low cost and high speed. Furthermore, we aim to examine how epigenetic modifier mutations shape the chromatin regulatory landscape (including histone modifications and chromatin accessibility) to promote clonal outgrowth in CM. We will use newly developed tools to assay genotype, chromatin accessibility and histone marks in ASXL1-mutant CH and FOXO1-mutant fatty liver disease to define differentially accessible and active regions, their associated genes and transcription factor binding motifs and their enrichment along lineage trajectories. By integrating these modalities, we will have an unprecedented view of the effect of somatic variants on the aging epigenome and gain insights into how these mutations may drive clonal expansion. This project will be ideal for a training physician-scientist, given its use of novel sequencing technologies and direct clinical implications. With the mentorship of my sponsor, thesis committee, MD- PhD program, and the support of this fellowship, I am confident I will be well prepared to pursue and achieve my goal of being a physician scientist and independent investigator.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY/ABSTRACT Viridans group streptococci (VGS) are the most common causes of bacteremia in neutropenic patients with hematologic malignancies and cause substantial morbidity and mortality. Fluoroquinolone (FQ) prophylaxis is recommended to prevent bacterial infections in neutropenic patients, but VGS that cause bacteremia despite FQ prophylaxis are FQ-resistant. There is an urgent need for novel approaches to prevent VGS infections in neutro- penic patients. The objective of this proposal is to determine how screening for colonization with FQ-resistant VGS could identify patients at high risk of developing VGS bacteremia despite FQ prophylaxis. An alternative approach to prophylaxis could then be pursued in these high-risk patients. The hypothesis is that neutropenic patients who develop VGS bacteremia despite FQ prophylaxis are colonized with FQ-resistant VGS prior to neutropenia and that microbial factors, such as the species of VGS and expansion of VGS in the oral microbiome, increase the risk of bacteremia in colonized patients. The specific aims of this project are: 1) Determine the optimal screening method to detect colonization with FQ-resistant VGS; 2) Determine the prevalence and clinical significance of colonization with FQ-resistant VGS in neutropenic patients and identify alternative prophylactic antimicrobial agents; 3) Identify risk factors for VGS bacteremia despite FQ prophylaxis in FQ-resistant VGS- colonized neutropenic patients. In this study, oral (buccal), oropharyngeal, and perianal swab specimens will be collected from 135 participants prior to hematopoietic cell transplantation (HCT). Specimens will undergo selec- tive culture to identify FQ-resistant VGS by direct plating and with a broth enrichment step to identify the optimal body sites to screen and the need for broth enrichment to detect colonization. The prevalence of and risk factors for colonization will also be identified. Study participants will be followed to determine how pre-transplant coloni- zation with FQ-resistant VGS increases the risk of VGS bacteremia during neutropenia. Colonizing and blood- stream FQ-resistant VGS isolates will undergo sequencing to determine whether patients develop bacteremia from their colonizing strain and antimicrobial susceptibility testing to identify alternative prophylactic agents with activity against these bacteria. Clinical and microbial risk factors for VGS bacteremia among colonized patients will then be identified, using 16S rRNA gene sequencing to assess the impact of the oral microbiome on this risk. The contributions of this proposal are that we will identify the optimal screening test to detect colonization with FQ-resistant VGS, determine the prevalence of colonization prior to HCT and the risk of VGS bacteremia in colonized neutropenic patients, and identify which colonized patients are at highest risk of VGS bacteremia. These contributions are significant and innovative because, if confirmed and expanded upon in a larger multi- center study, they could lead to a novel individualized approach to prevent VGS bacteremia where patients are screened for colonization with FQ-resistant VGS prior to neutropenia and their prophylactic approach is modified if colonization is detected, as opposed to the current “one-size-fits-all” approach of universal FQ prophylaxis.