Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2025 · 2024-02

ABSTRACT Merkel cell carcinoma (MCC) is a poorly understood cutaneous malignancy with viral etiology. Most MCC tumors feature monoclonal integration of Merkel cell polyomavirus (MCV), which expresses viral T-Antigens (T-Ags). Small T-antigen (sT-Ag) acts as a transcriptional co-regulator, while Large T (LT-Ag) principally functions to sequester retinoblastoma protein (RB1) to de-regulate the cell cycle. The T-Ags are responsible for driving tumorigenesis in T-Ag expressing (T-Ag+) MCC tumors: few mutations are present in T-Ag+ MCC tumors, including in tumor suppressors RB1 and TP53, whose activities are instead repressed by the functions of the T- Ags. Latent MCV infection that doesn’t result in T-Ag production is found in a substantial portion of the human population, however MCC occurs only rarely. Much of what occurs between latent infection and the appearance of a full-blown T-Ag+ MCC tumor has not been explored due to the lack of in vivo MCC tumorigenesis models. Among the unanswered questions is how the MCV T-Ags induce a tumor transcriptional program that features markers of multiple cell lineages, including both epidermal stem cell and neuroendocrine fates. Previous research in our laboratory determined that SOX9-expressing (SOX9+) hair follicle cells, which give rise to mechanosensory Merkel cells during fetal development, are also Merkel cell progenitors in adult skin. Hypothesizing that T-Ags reprogram cells in the Merkel cell lineage to cause MCC, I generated transgenic mice that express sT-Ag and ablate RB1 to mimic LT-Ag in SOX9+ cells. These mice developed tumors that were not bona-fide MCC but expressed neuroendocrine markers, making these mice a valuable model in which to study mechanisms of T-Ag mediated reprogramming. Analysis of tumors at early time points revealed that re- programming occurred only in specific sub-populations of SOX9+ cells and that reprogrammed cells were highly apoptotic. Therefore, I hypothesized that specific landscapes of gene accessibility are required for T-Ags to induce reprogramming from an origin cell and that suppressing p53 mediated apoptosis is required for MCC. I propose to leverage the model of SOX9-derived, T-Ag driven neuroendocrine tumors to generate novel insights into the mechanisms of T-Ag mediated reprogramming. I will use integrated epigenetic and transcriptomic sequencing analyses to characterize the gene accessibility and transcriptional landscape required for T-Ag mediated reprogramming to occur and identify a native cell type that is competent to undergo reprogramming. Furthermore, I will study how p53 suppression, which is commonly observed in MCC tumors, contributes to advancing reprogramming. Altogether, the proposed studies will establish a model of T-Ag mediated reprogramming in vivo and discover factors that enable T-Ags to reprogram cells. This research will not only make valuable contributions to the field of MCC research but also to my training by exposing me to molecular techniques and genomic and epigenomic analyses that are foundational to an independent research career.

NIH Research Projects · FY 2025 · 2024-02

Project Summary The number of physician-scientists in the United States has decreased from its peak of 5% in 1980 to 1.5% today. This decline particularly threatens Pediatrics and child health research where only 12.6% of all MD/PhD program graduates choose residency training in Pediatrics and in Internal Medicine, while 40% of U.S. MD/PhD graduates enter Internal Medicine residency programs, these trainees only account for 3.6% of all Internal Medicine trainees. Together, these data represent a critical need, and opportunity, for new and sustained efforts to reinvigorate the physician-scientist pipeline infectious diseases, allergy and immunologic diseases. The Icahn School of Medicine at Mount Sinai is well poised to answer this call for novel on-ramps into the Physician- Scientist Pathway during residency given its established infrastructure for Research Tracks in Residency which successfully track accomplished researchers to academic research-focused fellowships and are synergistic with this proposal. The opportunity to cultivate opportunities for research during residency is ideally suited to our robust translational research programs. Our excellence in biomedical research across the basic, clinical, and translational spectrum, along with the ability to individualize and adapt to each physician-scientist trainee, makes our environment an ideal place to train future physician-scientists. The Mount Sinai Stimulating Access to Research in Residency (StARR) is a new program to enhance physician-scientist research training opportunities related to the mission of the National Heart Lung and Blood Institute (NHLBI). Mount Sinai StARR will provide Internal Medicine and Pediatrics residents an entrez into the physician-scientist pipeline, especially focused on underrepresented minorities and residents with a passion for inquiry but who may not have had extensive research experience i.e. the “late bloomers”. The program will provide a stepwise introduction to investigative research to recruit resident trainees and will be an attractive recruiting tool across and outside the Mount Sinai Health System that will build an immersive individualized research experience with the goal of accelerating and retaining research independence for resident investigators.

NIH Research Projects · FY 2026 · 2024-02

Rare genetic diseases affect 3.5-6% of the population and are associated with diagnostic odysseys that can last up to decades. As first steps towards shortening diagnostic odysseys for infants and toddlers, we developed rules-based and natural language processing- (NLP-) based algorithms to identify infants and children aged 0–3 years who were typically ill. Our algorithms were accurate for identify atypical ill patients at these ages from electronic health records (EHRs). Cohorts so identified were strongly enriched for patients who had undergone genetic testing. Manual EHR review for such atypically ill patient who had never been evaluated for a rare genetic disease revealed that 52% could appropriately be referred for such an evaluation. During the UG3 phase, we will create a novel outpatient clinic, Mount Sinai Genetics Outreach (GO), staffed with medical geneticists with prior pediatric and internal medicine training, to evaluate patients identified by our EHR phenotyping algorithms. In a pilot study, we will deploy rules- and NLP-based algorithms to identify 200 children aged 0-12 years with >50% risk of having an undiagnosed rare genetic trait. We will survey pediatricians at five practices for baseline knowledge about diagnostic odysseys and genetic testing, provide education about the topic and then study the impact of our algorithm deployment. For patients referred to Mount Sinai GO, we will determine the outcomes of clinical genetic evaluations and diagnostic testing, including impact on subsequent health care. In order to improve our existing algorithms, we developed an automated abstraction engine that identifies patients diagnosed with 164 rare genetic disorders with 83% accuracy. We will expand this to more traits and use their EHR data to improve our pediatric EHR phenotyping algorithms. The goal is to increase sensitivity, currently at ~25%, without dropping precision below 50%. During the UH3 phase, we will deploy our optimized rare disease-detecting algorithms in a non-academic health care setting, Mount Sinai South Nassau Hospital, a non-academic community hospital setting without onsite medical genetic services. Our model will leverage pandemic-accelerated expertise in telehealth to facilitate access of underserved populations to genetics services. Our goal will be to achieve similar sensitivity and precision with our pediatric algorithms as well as a comparably successful referral mechanism. Also, we will extend our clinical rule-based and NLP algorithms to detect adolescent and adult patients likely to have rare genetic disorders and assess the impact of our approach on diagnostic odysseys. We will alter our pediatric rules-based algorithm, first to patients aged 12-21 years and then to younger adults. We will leverage our automated abstraction engine for rare genetic disease for iterative improvements. For adults, we will class traits by organ system in order to improve cohort size/statistical power. Finally, we will assemble and study information about diagnostic odysseys per se, including the impact of our algorithms in shortening them.

NIH Research Projects · FY 2026 · 2024-02

Project Description Immune cells are an integral part of the white adipose tissue microenvironment and have pronounced effects on the expansion and activation of thermogenic adipocytes, also known as beige/brite adipocytes. Anti-inflammatory factors are postulated to be beneficial for insulin resistance and diabetes. We recently reported that ablation of interleukin-10 (IL10)-associated anti-inflammatory signaling in mice, surprisingly, improved insulin sensitivity and glucose tolerance, protected against diet-induced obesity, and elicited browning of white adipose tissue. Yet, several questions remain about the mechanisms that mediate the metabolic actions of IL10: i) how does IL10 block adipose thermogenesis? ii) what are the fates of adipocytes deficient in IL10 signaling? and, iii) since IL10 is produced by multiple immune cell types, which adipose-resident immune cell is responsible for IL10 production? Our proposal builds upon our comprehensive data to address the crosstalk between immune cells and adipocytes within adipose tissue depots in regulating systemic metabolism and adipose thermogenesis. Our published and pilot studies show that interfering with IL10 activity in adipocytes is associated with increased mitochondrial respiration and energy expenditure. Reconstitution of wild-type bone marrow into IL10 knockout mice reversed the thermogenic phenotype. Ablation of adipose T cells increased adipose thermogenesis, pointing to a hematopoietic origin of the IL-10 signal regulating adipose tissue function. The IL10 receptor (IL10Ra) is highly expressed in mature adipocytes, and adipocyte-specific knockdown of IL10Ra decreases fat mass and increases adipose thermogenesis and energy expenditure. Conversely, IL10 treatment directly antagonizes the expression of thermogenic genes in a cell-autonomous manner. Furthermore, genome-wide Assay for Transposase-Accessible Chromatin (ATAC)-seq, ChIP-seq, and RNA-seq demonstrated that IL-10 represses the transcription of thermogenic genes by altering chromatin configuration at key enhancer and promoter regions through STAT3. Our findings identify the “IL10-IL10Ra-STAT3 axis” as a novel regulator of the thermogenic transcriptional program in adipose tissue and challenge conventional assumptions regarding links between immune and inflammatory signaling and adipose tissue function in obesity. To further test the hypothesis that adipocyte-specific IL10Ra directly “senses” IL10 in the microenvironment to limit adipose thermogenesis and energy expenditure, I propose three Specific Aims to define: i) mechanisms underlying IL10 inhibition of thermogenic gene transcription and beige adipocyte formation; ii) the fates of adipocytes in the presence and absence of IL10Ra, and iii) adipose resident immune cells responsible for anti-thermogenic activities.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY Many Parkinson’s patients develop cognitive impairment during an early, motor-asymptomatic pro- dromal period, before clinical diagnosis and motor symptoms. Cognitive impairment causes a significant disruption in quality-of-life for patients and caregivers. The onset of early cognitive impairment greatly increases risk for conversion to dementia, but such progression differs from that of other neurodegenerative diseases (like Alzheimer’s) and differs between males and females for reasons that are not clear. Cognitive abnormalities include impaired attention, slower information processing speed, and others, yet the underlying neurobiology of such early cognitive symptoms is not known, thwarting targeted therapeutic strategies which are necessary to halt or slow cognitive decline and dementia. Cognitive impairment is present in both idiopathic and hereditary forms of Parkinson’s, including that resulting from several mutations in LRRK2 that increase kinase activity. The focus of this project is on characterizing the underlying neurobiology of early cognitive deficits in a mouse model carrying a knockin G2019S gene mutation in LRRK2 that in humans increases risk of Parkinson’s-associated cognitive decline and dementia. The premise is built on our observations that young adult, male Lrrk2G2019S mice display significant deficits in attention (tested in a 5-CSRT task), slower information processing speed, and in fronto-striatal dependent instrumental learning. Such deficits could not be attributed to sensory perceptual deficits, differences in motivation or motor effects. Further, these deficits were normalized by systemic injection of the acetylcholinesterase inhibitor donepezil, implicating deficient cholinergic signaling. Anatomical analysis showed that the cholinergic innervation of mPFC was significantly sparser in male G2019S mice than in male wildtype controls. In contrast, young adult female G2019S mice showed normal instrumental learning, and an enhanced density of cholinergic innervation in mPFC compared to female wildtype controls, suggesting a compensatory response and demonstrating early sexually-dimorphic behavioral and anatomical outcomes driven by the mutation. We propose to examine cholinergic innervation of mPFC in male and female G2019S mutants across the lifespan and in the context of cognitive behavioral performance; we will interrogate projection-identified mPFC neurons by whole-cell electrophysiology to characterize cholinergic nicotinic and muscarinic receptor responses; and we will probe behaviorally-evoked ACh transients in mPFC using a genetically-encoded fluorescent ACh biosensor and will use this information to modulate cognitive function by optogenetically controling ACh signaling in mPFC during an attention task. Successful completion of these experiments will provide new insight into the earliest Parkinson’s-associated pathophysiology of mPFC cholinergic circuits and cognition, with the long-term goal to improve diagnostics, predict sex-specific clinical trajectories, and identify pathways for early precision therapeutic intervention.

NIH Research Projects · FY 2025 · 2024-02

Type 1 diabetes represents a major public health burden. While the availability of improved insulin pumps and continuous glucose monitors has eased some of the daily practical and psychological burden of T1D, technology has not dramatically improved glucose control, with studies conducted by the T1DExchange ~5 years apart showing no improvement in HbA1c over the lifespan. There has also been only limited improvement in the shortened lifespan observed in those with T1D, and the incidence of T1D has increased over the last twenty or more years. Initiated and perpetuated by a complex interplay of genetic risk factors, environmental triggers and dysregulated immune responses, the natural history of T1D unfolds in a series of events that progress at variable rates towards onset of clinical disease. Yet the precise correlates and determinants of T1D pathogenesis remain incompletely defined. The identification and validation of novel biomarkers is critical to promote better stratification of T1D risk and progression, can illuminate pathways of disease pathogenesis and heterogeneity, and likely will inform the development of interventional strategies to delay or prevent disease. To precipitate the identification of robust and improved biomarkers, the TrialNet consortium designed the key question 1 (KQ1) PBMC study, a nested case/control study of 178 multiple Aab+ at-risk individuals that over a period of three years either did (“cases”) or did not (“controls”) progress to clinical disease. This earlier study aimed to correlate the evolution of transcriptomic and phenotypic immune cell profiles with incipient T1D onset. KQ1 analyses incorporating detailed clinical metadata are currently ongoing, and we now seek to complement these efforts with a broad interrogation of soluble serum factors from the very same KQ1 donor cohort. Here, the “KQ1 serum study” aims to 1., to identify and validate serum biomarkers for T1D risk, development and/or progression; 2., to establish a comprehensive data resource for reference and orientation, hypothesis generation and improved interventional study design; and 3., to execute broadly integrated data analyses in particular with emerging transcriptional and phenotypic immune cell data from the KQ1 cohort. To this end, we have identified and prioritized suitable targets and analytical approaches, robust technology platforms, and investigator teams with track records to perform high-throughput analyses. Since the scope of our proposal is delimited by the amount of available KQ1 serum samples, we developed a highly collaborative Research Plan that deploys both discovery- and hypothesis-driven investigative strategies. Our proposed interrogations include expansive measurements of serum proteins, antibody responses against commensal bacteria, PBMC responder transcriptomics, and exocrine pancreatic factors. Integrative analytics will be leveraged to define novel serum signatures for T1D risk stratification, improve prediction of disease progression rates, and elucidate aspects of T1D pathogenesis that may accelerate the development of tailored diagnostic, prophylactic and therapeutic modalities. Altogether, the KQ1 serum study has unique potential to build a robust framework for future integration of serum biomarkers in the improved management of T1D.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY Obstructive Sleep Apnea (OSA) is a common chronic condition that affects more than 1 billion people worldwide. Currently, presence of OSA and its severity is assessed using the apnea-hypopnea index (AHI) that measures the frequency/rate of respiratory events. AHI has demonstrable limitations including lack predictive accuracy and poor relationship to outcomes. Task forces across the American Academy of Sleep Medicine, Sleep Research Society and others internationally have made an urgent plea to move the diagnosis and management of sleep apnea beyond the AHI. In response to this, our group has recently developed a physiology guided AI/ML approach that utilizes the known OSA pathophysiology across ventilatory, hypoxic, and arousal burdens and combines them into probability scores that provide risk of short- and long-term adverse outcomes of OSA. We use fully automate, reliable, and interpretable measures for ventilatory, hypoxic, and arousal burden towards our pursuit of explainable AI. Our preliminary data from 10,952 subjects across three epidemiological cohorts suggests that the machine-learned sleep apnea probability of sleepiness (SAPs), which combines ventilatory/hypoxic/arousal burdens, classifies sleepiness with accuracy of 87%. Further, using the same approach but with different set of weights that were trained to predict cardiovascular (CVD) and all-cause mortality, the sleep apnea probability of CVD (SAPcv) predicted CVD and all-cause mortality across 4,784 subjects with an accuracy of 81% and 88% respectively. To be translatable into clinical practice, validation of AI/ML approaches in a sleep clinic population is needed and is the core objective of our proposal. In Aim 1 we will validate SAPs and SAPcv as probability scores for risk of short- and long-term outcomes across a sleep clinical population comprising data from a diverse set of subjects (N=6,393) seen at the Mount Sinai Health System from the Greater New York Area. In Aim 2, we will assess real-world performance of SAPs across subjects (N=700) see at Mount Sinai, as a tool in the decision-making pipeline to predict improvements in sleepiness with 3 months of treatment using any modality (e.g., CPAP, oral, positional etc.). Finally, in Aim 3, we will use causal random forests to assess conditional average treatment effects, thereby identifying subgroup of subjects who are likely to benefit from treatment with respect to CVD outcomes. Our proposal will offer crucial evidence needed to translate metrics that go beyond AHI in assessing severity of OSA into clinical practice and are thus poised to shift the paradigm in clinical management of OSA.

NIH Research Projects · FY 2026 · 2024-02

Chronic obstructive pulmonary disease (COPD) affects 10-15% of U.S. adults, costs over $37 billion annually, and causes serious symptoms, physical impairment and high hospital admission and mortality rates. Much of its burden can be reduced by following guideline recommended self-care activities, like daily use of disease controlling medications and pulmonary rehabilitation. Yet patients often struggle with the self-care needed to maintain or improve their health. The potential barriers to successful self-care are numerous, encompassing socioeconomic, psychosocial, physical, cognitive and other domains. Moreover, many of the barriers are major contributors to poor health outcomes among Black and Latino patients. Most COPD self-management support interventions have largely followed one-size-fits-all strategies of education and training. We created the Supporting self-Management Behaviors for aging Adults (SaMBA)-COPD intervention based on the successful SaMBA program for older adults with asthma. The model is unique among chronic illness self-management programs because it involves comprehensive screening for barriers to self-management and disease control and couples it with tailored, theory-based, barrier-specific interventions. The intervention is delivered by community health workers and includes steroid and antibiotic prescribing for early intervention of COPD exacerbations and home-based pulmonary rehabilitation (HBPR). Our feasibility trial of SaMBA-COPD, conducted with a predominantly Black and Latino population, met most of the benchmarks for success, including complete preparation of all intervention and study related materials and protocols, intervention fidelity and patient acceptability, and outcomes that suggest the intervention will improve self-management behaviors and reduce COPD symptoms. We now propose a fully powered randomized trial among inner-city adults with COPD (n=300). The Specific Aims are to test the impact of SaMBA-COPD on (1) COPD controller medication adherence and other self-management behaviors at 6 months; (2) on clinical outcomes (COPD symptoms measured with the COPD Assessment Test and 6-minute walk test distance) at 6 months, and emergency department visits and hospitalizations at 12 months; and (3) to examine mechanisms of intervention effect on medication adherence and clinical outcomes, with a focus on the role of COPD illness and medication beliefs. We will recruit adults ages 40 years and older with advanced COPD in outpatient practices in New York City. The intervention will be delivered for 6 months. Attention control patients will receive untailored COPD education. Sustainability of treatment effects will be evaluated at 12 months. The intervention is significant and innovative for its comprehensive pre-trial preparation, attention to the full range of contributors to suboptimal self-management and COPD control, the broadening of community health workers’ scope of practice to include HBPR, and the project’s focus on populations with high rates of poor outcomes in COPD.

NIH Research Projects · FY 2025 · 2024-01

SUMMARY Regulatory T cells (Treg) are critical in controlling alloreactive T cell responses in organ transplant recipients, but differences in suppressive capacity exist within phenotypically similar cells. Despite transcriptional heterogeneity within the Treg compartment, lack of available technical approaches has prevented any assessment of their functional heterogeneity. The Treg suppressive capacity is associated with a distinct metabolic signature defined by increased oxidative phosphorylation (OXPHOS) and decreased glycolysis. Cellular reduction-oxidation (redox) homeostasis mechanisms are fully intertwined with metabolic state; T cells engage specific metabolic pathways and also rearrange their redox system, to maintain T cell metabolic rewiring and cell fate. Regulation of de novo synthesis of the antioxidant glutathione (GSH) is a crucial redox homeostasis mechanism in Treg. These cells respond to activation-induced oxidative stress by increasing glutamate cysteine ligase (GCL) activity to form gamma-glutathione cysteine (-GC) and maintain high GC and low GSH intracellular levels (while Teff then shuttle -GC to increase intracellular GSH). This particular balance of high -GC and low GSH (GSH homeostasis) needs to be preserved for Treg to exert a suppressive function. The current techniques for capturing GSH homeostasis in Treg are inadequate and the only available assays measure -GC in cell lysates (not in live cells). In addition, most commercial GSH sensors bind irreversibly to GSH, eliminate it from the cells and thus completely disrupt GSH homeostasis and lose information on heterogeneity, which we contend is crucial. In this proposal, we will develop an approach to capture the heterogeneity in GSH homeostasis in Treg using a small-molecule dual -GC/GSH sensor that overcomes these limitations. We will use it to test the hypothesis that cell-to-cell differences in GSH homeostasis within Tregs underlies the heterogeneity in Treg suppressive capacity (Aim 1), and to provide proof-of-concept that this approach can capture clinically relevant heterogeneity in circulating Treg from kidney transplant recipients treated with anti-IL-6R antibody (tocilizumab) (Aim 2). At the end of the R21 we will have generated a small-molecule biosensor to obtain GSH-homeostasis profiles in Treg, mapped the relationship onto Treg suppressive capacity, and demonstrated the translational potential of this approach. This is a technically challenging approach (high-risk), but the results have the potential to provide a tool to identify previously undetected functional differences within the Treg compartment (high- reward), setting the basis for an R01 testing the mechanisms responsible for this heterogeneity.

NIH Research Projects · FY 2026 · 2024-01

The search for an HIV cure has been among the highest HIV research priorities for several decades. While much knowledge was gained regarding the persistent HIV reservoir, HIV eradication remains elusive. There is an emerging consensus that the HIV reservoir is not a fixed assortment of epigenetically silenced proviruses but rather a dynamic compendium of mostly defective proviruses. We recently showed that most infections in primary human CD4+ T memory cell subsets result in non-productive, latent infections. These findings were made possible thanks to our newly developed HIV vector system, which permits distinguishing and separating primary human cells harboring non-productive and productive proviruses. Transcriptome analysis of specific CD4+ T cell subsets identified transcript isoforms differentially expressed in the context of productive and non- productive infections. We propose an integrated experimental approach that will systematically dissect non- productive infections in primary human CD4+ T cells. First, we will analyze the proviral structure of non- productive and productive infections (Specific Aim 1), which will include both integration site analysis as well as detection of defective proviruses. We will identify and validate the transcriptional programs specific for productive and non-productive infections using advanced bulk and single cell long read transcriptome analysis approaches (Specific Aim 2). We will validate findings using genome editing tools and a panel of viral mutants lacking functional accessory genes. Lastly, we will dissect the role of innate immune responses in the context of non-productive infections (Specific Aim 3) by determining to what extent stimulation with gamma cytokines or Type-1 interferon changes the propensity of CD4+ T cell subsets to support non-productive infections. Collectively, the proposed studies will deliver insights into the interplay between non-productive HIV infections and host cell environment pointing to the molecular mechanisms that drive non-productive infections in primary human CD4+ T cells. The results obtained will also provide essential information on potential HIV latency biomarkers and directly inform of innovative HIV cure strategies.

NIH Research Projects · FY 2025 · 2024-01

My long-term goal is to integrate health informatics, data mining and machine learning to improve the care for patients with, and at risk for, acute kidney injury (AKI). I am dual trained in Nephrology and Critical Care Medicine. I am already developing my skills in health informatics. This proposal presents a five-year career development plan for NIH K08 award focused on training in advanced data mining, machine learning and their applications to critical care nephrology. To that effect, I have assembled a strong mentoring team with decades of experience in mentoring, research and leadership. The outlined career development plan in conjunction with intensive mentoring and hands-on training will provide me the perfect platform to become a leading independent investigator in the field. AKI is seen in over one-third of patients undergoing cardiac surgery. Several trials investigating various medications to prevent or treat AKI over the last two decades have proven futile. Management of AKI therefore focuses on its prevention, measures to reduce further progression and management of its complications. The strategy to prevent AKI and its progression relies on clinical interventions to optimize a patient’s fluid status, blood pressure and avoiding nephrotoxins and hyperglycemia. These clinical interventions when provided to patients requiring cardiac surgery as a care-bundle are associated with decreased incidence of AKI. This care- bundle, however, has very low compliance with implementation and lacks the ability to personalize care for patients. With prior work showing differential response to therapy in AKI phenotypes, there is a critical need to determine personalized strategies to prevent the development of persistent AKI. Personalization of treatment strategies based on dynamic clinical characteristics of patients will ensure that the right action is performed at the right time. As transient AKI resolves spontaneously within 48 hours, focusing interventions to those at high risk for developing persistent AKI will lead to further personalization of this approach. The overall objective of this project is to determine a personalized strategy using machine learning to prevent the development of persistent AKI after cardiac surgery. I will pursue following specific aims for this study: (1) Develop reinforcement learning (RL) based strategy to prevent the development of persistent AKI after cardiac surgery. (2) Develop digital biomarkers to predict patients at risk for persistent AKI after cardiac surgery. Completion of these aims will provide a structured framework to provide personalized care to prevent the development of persistent AKI after cardiac surgery. It will also provide me with preliminary data and experience necessary to apply for R01 applications as an independent investigator leading a data science research program in critical care nephrology.

NIH Research Projects · FY 2025 · 2024-01

The goal of the project is to identify pre-diagnosis plasma biomarkers for AMD, i.e., markers that can be measured in plasma samples from healthy donors and may predict which donors will get AMD in the following years. We will use approx.150 plasma samples collected and stored at the Biome MSSM Biorepository, from healthy, non-AMD donors, which eventually became diagnosed with AMD 2-8 years after the blood donation. These samples will be compared to samples from donors that did not develop AMD after a similar number of years. We will use two strategies. The first strategy (Aim 1) consists of analyzing the protein composition of plasma samples using the SOMAscan platform, which allows quantitation of approx. 7000 proteins. In addition, we will investigate whether pre-diagnostic differences exist in the optical spectra generated by Fourier Transform Infrared (FTIR) spectroscopy. Aim 2 is based on the hypothesis that some post-diagnostic biomarkers, shown by published studies to robustly differentiate healthy donors from AMD patients, reflect differences that pre-exist years before the donors are diagnosed. We will investigate, using the BioMe plasma samples, whether the differences exist years before the disease develops. As most BioMe donors have been fully genotyped, genetic information will be also included in the effort to find pre-diagnostic biomarkers. Regarding the potential impact of the study, a large-scale effort is underway for years to find dietary supplements and other agents that could prevent AMD. Such studies involve administration for years of supplements to relatively large numbers of subjects that do not have yet AMD and waiting for years to determine if the proportion of persons that develop the disease is reduced. Development of predictive biomarkers could enormously simplify such efforts, by assessing the immediate effect of the supplements on the biomarkers. At the level of individual subjects, if protective interventions are found, the pre-diagnostic markers will help detect the persons for which the interventions are necessary.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY In 2020, 53 million people in the U.S. served as family caregivers, the partners, relatives, and friends who provide assistance (i.e., physical, emotional) to patients with often life-threatening, incurable illnesses. Caregivers are increasingly tasked with responsibilities once performed by medical professionals, and the availability and health of supportive caregivers is more critical than ever. A growing number of caregivers provide care to patients with advanced, life-limiting cancers, and are tasked with critical patient care responsibilities, and play a significant role in healthcare communication and advanced care planning as healthcare proxies. The burden of these responsibilities is great and is driven largely by existential distress, which contributes to anxiety, depression, poor quality of life, and mental health challenges in bereavement. Concurrently, caregiving is an opportunity to experience a profound sense of meaning and purpose; caregiving allows for the realization of new strengths and capacities, healing of relationships, and refinement of life goals. When existential distress is addressed, caregivers can experience an enhanced sense of well-being despite their challenges. While many interventions have been developed to support cancer caregivers, none directly target existential distress. Our group adapted Meaning-Centered Psychotherapy (MCP), a highly effective intervention in decreasing existential distress and enhancing well-being among patients with advanced cancer, for cancer caregivers. Meaning-Centered Psychotherapy for Cancer Caregivers (MCP-C) is a stakeholder- informed, innovative, manualized intervention designed to assist caregivers to connect to meaning and purpose in life, despite the challenges of caregiving. Results of our pilot randomized controlled demonstrated MCP-C’s feasibility, acceptability, and superiority in improving meaning, benefit-finding, depression, and spiritual wellbeing. In the proposed trial, we will more rigorously evaluate the efficacy of MCP-C through a randomized controlled trial of 200 caregivers of patients with advanced (Stage III/IV solid tumor) cancer who will receive 7 sessions of MCP-C or Supportive Psychotherapy for Caregivers (SP-C), the standard of community-based caregiver care. Participants will undergo assessments of meaning and spiritual wellbeing (primary outcomes), and anxiety, depression, benefit finding, caregiver burden, and social support (secondary outcomes) at baseline, post-treatment, and at 6- and 12-months follow-up. Bereavement outcomes including pre- and post-loss grief, preparedness for loss, and regret will also be evaluated at each time point. We predict MCP-C will result in greater improvements in primary and secondary outcomes, and that sense of meaning in life will mediate treatment effects. We also predict that MCP-C will result in better preparedness for loss and improved pre- and post-loss grief and regret. Our results will enhance our capacity to powerfully target existential distress in caregivers of patients with advance cancer and by extension, improve their capacity to provide critical care to patients at end-of-life.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY: “Targeting the Androgen Receptor in Bladder Cancer to Enhance Sensitivity to Immune Checkpoint Blockade”. Up to 70-80% of patients with bladder cancer demonstrate de novo resistance to PD-1/PDL-1 immune checkpoint blockade (ICB). Of those patients that initially respond to ICB, most ultimately develop resistance [1- 5]. These data support the investigation of additional cooperative targets which may sensitize bladder cancers to immunomodulatory therapies. Androgen deprivation therapy (ADT) is the standard of care treatment for prostate cancer patients. However, ADT has also proven to delay progression of AR positive bladder cancers and, thus, offers a potentially viable co-target for enhancing the activity of clinically approved PD-1/PDL-1 targeting agents. Emerging evidence in prostate cancer supports an AR-dependent mechanism regulating ICB response whereby T cell intrinsic expression of AR blocks T cell activity and differentiation. We hypothesize that clinically approved ADTs, including gonadal castration and AR targeted inhibitors, will promote the function of tumor infiltrating T cells while concomitantly inhibiting androgen sensitive epithelia in bladder tumors. We have developed preclinical mouse models and techniques that will test the impact of ADT on early and late-stage bladder cancer progression. We will leverage established methods in single cell transcriptomics to understand the mechanism(s) of how ADT could alter the immune microenvironment to sensitize tumors to ICB. To further understand the cell intrinsic role of AR, we will leverage an established preclinical model in the Mulholland laboratory to genetically disrupt AR expression in specifically in T cells. Our team. We will use an effective collaboration between experts in the field of cancer modeling, oncology, urology, bioinformatics and statistics. The translational research team consists of: David Mulholland, PhD (preclinical mouse modeling), Matthew Galsky, M.D. (GU oncology), John Sfakianos M.D. (surgical urology), Ernesto Guccione, PhD (bioinformatics), and Madhu Mazumdar, PhD (biostatistics). Impact. If successful, our exploratory studies will generate data that is highly impactful for bladder cancer patients who are poorly responsive to standard of care immunomodulatory therapies.

NIH Research Projects · FY 2026 · 2023-12

Summary The RAS/RAF/MEK/ERK (RAS/MAPK) signaling pathway is an established driver of many cancers, commonly hyperactivated by genetic alterations in RAS or BRAF. RAF and MEK inhibitors (RAFi and MEKi) have been clinically successful in BRAF-mutant (BRAF-MUT) tumors, but their clinical efficacy in RAS-mutant (RAS-MUT) tumors has been so far modest, limited by toxicities. Based on the experience with BRAF-MUT tumors, we posit that clinical efficacy of RAS pathway-targeting inhibitors requires selective MAPK inhibition in RAS-MUT tumors over RAS-WT normal cells. To achieve such RAS-MUT tumor-selective potency of RAS pathway inhibitors, we need better understanding of how RAS activation affects target drug binding and MAPK inhibition in cells. The goal of this proposal is to accomplish a better understanding of how the cellular context affects tumor-selective therapeutic potency of RAFi and MEKi, with the goal of designing more effective therapeutic strategies for RAS- MUT cancers. Our preliminary findings suggest that RAS activation differentially affects RAFi binding to RAF paralogs, as well as their protein stability. We have further evidence that although RAFi suppress MAPK preferentially in RAS-MUT over RAS-WT normal cells, combinations of RAFi and MEKi result in similar MAPK inhibition in the two contexts, they are thus predicted to promote dose-limiting toxicities. Our hypothesis is that RAS activation affects protein expression levels and conformations of RAF paralogs and MEK resulting in differential binding to RAFi and MEKi, determining tumor-selective MAPK and cell growth inhibition by these drugs. This hypothesis will be tested in three Aims: Aim 1 will characterize differential RAFi binding to RAF paralogs in RAS-MUT and normal cells using in-cell binding assays and structural and biophysical approaches. Aim 2 will define effects of RAS activation on RAF paralog protein stability and expression. Aim 3 will characterize biochemically, biophysically and in vivo combinations of diverse RAFi and MEKi to determine whether the lack of tumor-selective suppression of MAPK signaling in RAS-MUT tumor over RAS-WT normal cells increases toxicity and limits the efficacy of combined RAFi+MEKi. The proposed research will develop experimental tools and concepts to delineate mechanisms determining differential potency of RAFi and MEKi in RAS-MUT over normal cellular contexts. Defining such mechanisms should enable the rational design of more effective RAS pathway-targeting drugs and drug combinations for the treatment of RAS-MUT cancers.

NIH Research Projects · FY 2025 · 2023-12

Asthma affects 8% of the United States population >60 years and causes considerable harm: older adults are 4 times more likely to die from asthma and have twice the risk of hospitalization. The burden of asthma is notably greater among minoritized older adults. Research suggests that perception of expiratory airflow limitation may be a major determinant of asthma outcomes in older adults, and that older adults are substantially less aware of airway obstruction than younger adults. These observations suggest that perception of airflow limitation is a potential target for improving outcomes of older patients with asthma; however, there has been very limited research in this area. We completed a pilot randomized controlled trial (RCT) of an intervention that trains older adults with asthma to better perceive expiratory airway obstruction through feedback via peak expiratory flow (PEF) prediction and couples this feedback with motivational interviewing (MI) to promote change in asthma self-management behaviors. Compared to an attention control, the intervention improved PEF, perception of airflow limitation and asthma control. In this project, we propose to conduct a fully powered RCT to test the intervention’s efficacy in a large cohort of older adults with asthma and understand the mechanisms underlying improvements in asthma control and objective measures of expiratory airflow. The Specific Aims are to (1) Test the impact of the intervention on perception of expiratory airflow limitation in older adults with asthma, (2) Examine the efficacy of the intervention for improvements in lung function (forced expiratory volume in one second [FEV1] and PEF), self-reported asthma control (Asthma Control Questionnaire [ACQ] scores), quality of life (Asthma Quality of Life Questionnaire [AQLQ] scores), and emergency department and hospital use, (3) Test the intervention’s impact on mean daily ICS dose used (daily maintenance and as needed) and illness beliefs. We will conduct a RCT of 300 older (≥60 years) patients with uncontrolled asthma who are on controller medications (daily maintenance or as needed) recruited from underserved inner-city practices and randomize them to an intervention consisting of PEF feedback training and MI (with or without a booster session at 6 months) or a time-matched attention control group. The intervention will be delivered for 3 sessions over 6 weeks. Control patients will receive supportive counseling related to their asthma and standardized, untailored asthma education during 3 individual sessions. Data will be collected at baseline, 1-month, 6-months (primary analyses of effectiveness) and 12-months post- intervention. In secondary analyses, we will test the sustainability of treatment effects with vs. without the booster treatment session (active booster vs. attention control booster) delivered immediately after the 6 month assessment on outcomes at 12-months. The project addresses a significant public health problem in a vulnerable patient population. The innovative intervention addresses a significant public health problem among patients who are rarely the focus of interventions to improve asthma outcomes.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY / ABSTRACT Pluripotent stem cells hold great promise for regenerative medicine. Beyond their ability to restore normal tissues, their potential can be expanded even further through engineering approaches to create new lineages with therapeutic properties that are not normally made in vivo. In preliminary studies, we have created pluripotent stem cell lines that 1) have been genomically edited such that they are able to cross human-mouse barriers; 2) engineered these cells to carry a knockin insertion of a broadly neutralizing HIV antibody at the endogenous loci. With these cells, we will develop strategies to differentiate human pluripotent stem cells into transplantable pathogen-specific plasma cells. This process will involve iterative and recursive steps to engineer pluripotent stem cell differentiation to downstream hematopoietic, B cell, and plasma cell lineages using single cell transcriptional trajectories as guides. These targeted approaches will be complemented with unbiased genetic and small molecule screens for new pathways that promote or inhibit specific steps in differentiation, and additional single cell transcriptomic and epigenetic analyses to provide templates for cellular engineering.

NIH Research Projects · FY 2026 · 2023-12

Stroke is a leading cause of disability and mortality with risk stratification based on a variety of environmental and social factors. The New York City Collaborative Regional Coordinating Center (NYCC-RCC) will unite three complementary research-intensive biomedical institutions (Einstein, NYU, Mount Sinai), three organizationally linked partners, and a network of high-volume clinical performance sites to further StrokeNet’s mission and to reduce the disproportionate burden of stroke in vulnerable populations. The NYCC-RCC has a record of leadership in community engagement and partnerships that will facilitate high-risk and understudied patients’ enrollment in StrokeNet trials. The NYCC-RCC has significant expertise in the conception, development, and execution of clinical trials. The revamped NYCC-RCC will utilize a number of key design features to fulfill StrokeNet’s mission, including: 1) Organizational collaboration through the integration of three leading academic health systems with targeted organizationally linked partners and clinical performance sites enhancing specific focus areas; 2) Recruitment and retention of representative populations through community engagement and strategic partnerships; 3) Focus on vulnerable pediatric stroke patients by integrating established pediatric stroke leadership with centers of excellence treating diseases known to confer high stroke risk; 4) Coordination of multiple high-volume thrombectomy centers to facilitate STEP platform enrollment; 5) Establishment of programs/infrastructure to facilitate hyper-acute stroke trial participation; 6) Complementary, multidimensional MPI structure including an imbedded career development program; 7) A structured trial development process coupled with senior trialist guidance to generate a pipeline of StrokeNet trial sub-missions; 8) Pre-clinical expertise in evolving translational areas, including epigenetic and regenerative/stem cell therapeutic applications; 9) RCC connectivity via an advanced information technology and network performance group (IT/NPG). The NYCC-RCC will employ an integrated mentorship model to ensure career development of promising stroke investigators, including a structured education plan for StrokeNet fellows utilizing the breadth and sophistication of our CTSA’s educational curriculums. The NYCC-RCC will train junior and mid-career investigators, as well as clinical research coordinators, toward productive careers in stroke research. In summary, the NYCC-RCC will leverage established clinical trials expertise with extensive scientific and organizational excellence to fulfill the StrokeNet’s mission. Therefore, this proposal seeks funding for an NIH StrokeNet – Regional Coordinating Stroke Center U24 – RFA-NS-23-010.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY Suicide results in 130 preventable American deaths every day, and >700,000 deaths worldwide each year. Non- fatal suicide attempt (SA) affects an even larger proportion of the population with estimates 10-25 times the number of individuals who die by suicide (1.4 million in the US, >20 million worldwide). Suicidal ideation (SI), the contemplation of taking one’s own life, is even more common, with a cross-national lifetime prevalence of 9.2%. While these suicide outcomes are all significantly heritable, only recently have samples reached sufficient size to conduct well-powered genetic studies, and thus far, these have largely been limited to SA. Large-scale genetic studies of the full spectrum of suicide outcomes are necessary to elucidate their genetic and biological etiologies, potential drug targets, and the similarities and differences between them. Here, we establish the Suicide Working Group of the Psychiatric Genomics Consortium, to interrogate the biological basis of the full spectrum of suicide outcomes in individuals of diverse ancestries, leveraging clinical, population, and medical examiner resources worldwide. This project will characterize the genetic etiology of suicide outcomes through genome-wide association studies of at least 69,800 SA cases, 19,500 suicide death (SD) cases, and 206,900 SI cases. We will elucidate the shared and distinct genetic etiology between suicide outcomes and psychiatric disorders, and between SA, SD and SI, illuminating the similarities and differences between them. These results will be used to identify biologically relevant tissues, cell-types, pathways, and drug targets, and to prioritize causal genes and SNPs underlying genome-wide significant loci. Overall, this proposal dramatically expands and diversifies efforts to understand the etiology of suicide outcomes, combining prior efforts and expertise from the PI and Co-Is in genomic studies of SA, SD and SI. The study will provide novel biological insights into genetic risk of suicide outcomes that will enable potential avenues of therapeutic understanding and risk stratification. Finally, it will facilitate the worldwide collaborative growth of a highly-powered cohort of diverse ancestries, environmental exposures, and phenotypes, in which to address many future questions related to suicide outcomes.

NIH Research Projects · FY 2026 · 2023-12

Project Summary/Abstract Infection of the olfactory epithelium in COVID-19 patients is thought to underlie loss of smell (anosmia), a pathognomonic symptom that can be long-term in some patients, significantly affecting quality of life. The olfactory epithelium is also believed to be a major entry point for systemic SARS-CoV-2 infection, which can result in neurological as well as respiratory symptoms. Wild-type SARS-CoV-2 cannot bind the mouse ACE2 receptor, and existing human ACE2 (hACE2)-expressing mouse models either do not permit conditional analysis or do not confer severe illness after infection. Due to the lack of genetically manipulable models that display severe disease, the infected cell types responsible for acute and long-term anosmia, and the route(s) by which the virus penetrates the brain, have not been definitively identified. To address these gaps in knowledge and test cell type-specific requirements for COVID-19-related pathologies we generated conditional hACE2fl knockin mice that express hACE2 in similar cell types to humans. hACE2fl mice nasally inoculated with a high dose of wild-type SARS-CoV-2 display initial infection of olfactory epithelium and rapidly develop anosmia. This is followed by infection of neurons in the olfactory bulb and brain, which is associated with lethality and requires neuronal hACE2 expression. Importantly, specific destruction of olfactory epithelium via methimazole treatment prevents olfactory bulb and brain infection and lethality, identifying the olfactory epithelium as an essential gateway to CNS infection. hACE2fl mice inoculated with a low dose of wild-type SARS-CoV-2 show reversible disease and survive, but a subset displays a long-term decrease in odor sensitivity (hyposmia) like that observed in humans. We propose to use hACE2fl mice to provide definitive genetic evidence for cellular mechanisms of short- and long-term loss of smell and identify the pathways for brain infection during COVID-19. These studies are expected to complement existing descriptive human studies to identify causal pathogenic mechanisms and preventative and therapeutic targets. Three specific aims will be pursued: (i) define the cellular requirements for acute loss of smell; (ii) determine the mechanisms of long-term hyposmia; and (iii) uncover the cellular mechanisms of olfactory bulb and brain infection.

NIH Research Projects · FY 2026 · 2023-12

Persons with multiple sclerosis (MS) suffer neurodegeneration and functional decline despite effective management of new inflammatory lesion formation by modern treatments. We must therefore identify modifiable risk and protective factors associated with disease progression. There is high interest among people living with MS and their clinicians about the protective effects of diet, and we have linked Mediterranean diet to less cerebral atrophy and better functional outcomes, but the underlying mechanisms are unknown. This R01 will evaluate biological aging as a mediator for observed effects of diet on neurodegeneration and disease progression in MS. Our team has linked biological aging to neurodegeneration in MS, and others have linked biological aging to Mediterranean-style diets in non-MS populations, which informed our hypothesis that biological aging (BA), measured through the National Health and Nutrition Examination Survey Biological Aging Index (NHANES BAI) and leukocyte telomere length (LTL), is a mediator of observed effects of diet on neurodegeneration in MS. The study will leverage two distinct pre-existing cohorts with rich clinical and imaging data and specimen collection. The first is RADIEMS (Reserve Against Disability in Early MS), an NIH- funded longitudinal study of risk and protective factors for disability in people with early MS that enrolled 185 people with MS in 2016-2017. This cohort provides longitudinal dietary, clinical, and imaging data, and specimens. The current proposal will link into RADIEMS follow up for the next 3 years but will also be able to powerful analyses encompassing 8 years of follow up data. The second is a cross- enrolled in Project Y in 2016 at age 50. The Project Y cohort will provide additional information on an older population of individuals with MS as well as provide a data set where chronological age is constant, simplifying the biological aging analyses. The specific aims are designed to evaluate: the effect of diet on biological age in people with MS (aim 1), the impact of diet and BA on brain atrophy and the degree to which dietary relationships are mediated by BA (aim 2), the impact of diet and BA on clinical MS worsening and the degree to which dietary relationships are mediated by BA (aim 3). These will be accomplished by adding the NHANES BAI to RADIEMS (which requires participant interaction but can be done because RADIEMS is ongoing) and by adding LTL analysis to both cohorts (which can be done using banked samples). Detailed data on other health behaviors and demographics are available for both cohorts and will be considered and accounted for in all analyses. This study will contribute to a growing body of literature that will help inform a more holistic approach to MS care, which is eagerly sought by people living with MS. The results of this proposal have high potential for impact in advancing our understanding of the pathophysiology of neurodegeneration and informing recommendations for lifestyle modifications to promote neuroprotection in MS.

NIH Research Projects · FY 2024 · 2023-12

SUMMARY Itch is described as an unpleasant sensation that elicits the desire to scratch. Although acute scratching is the protective mechanism to remove irritants from the skin, chronic itch is debilitating. In addition, chronic itch is widespread and very difficult to treat because of a lack of understanding of the underlying mechanisms. Therefore, it is critical to gain a better understanding of the cellular and molecular basis of chronic itch toward the development of novel and effective therapies. Despite great progress in the past few decades in unraveling the role of membrane bound G-protein coupled receptors and ion channels, especially transient receptor potential (TRP) channels in the generation of itch sensation at the levels of primary sensory neurons and spinal cord, much remains unknown about how the cells and molecules in the skin contribute to the production and regulation of chronic itch other than the mediation of allergic itch by mast cells. Pilot studies showed that skin-specific knockout of Piezo2 severely reduced the spontaneous scratching in multiple mouse models of chronic itch. Moreover, the reduction of spontaneous itch in mice subjected to experimental dry skin is correlated with a loss of mechanically evoked C-fiber firing mediated by the TRPV1- positive C-mechanoreceptors. We thus hypothesized that Piezo2-Merkel cell signaling is required for the generation of spontaneous itch under chronic itch conditions by driving the TRPV1-positive C- mechanoreceptors under chronic itch conditions. In this grant proposal we will: 1) Use unique genetic approaches to investigate in vivo functions of the mechanosensitive Piezo2 channels and mechanosensory Merkel cells in mediating spontaneous itch three well-established mouse models of chronic itch; 2) Demonstrate that miswiring occurs between the Merkel cells and the pruriceptive C-type mechanoreceptor to promote spontaneous itch in chronic itch mice, thereby providing the cellular and molecular basis for chronic itch in multiple mouse models of chronic itch. Our findings will provide a major contribution to our general understanding of how Piezo2 channels and Merkel cells affect itch signaling in the skin, and undoubtedly lead to new therapeutic approaches for treating chronic itch.

NIH Research Projects · FY 2026 · 2023-11

PROJECT SUMMARY In all animals, including humans, establishment and maintenance of the germline stem cells (GSCs) is critical for life-long reproductive health and fitness. Yet, how GSCs are specified in vertebrates remains largely unknown. GSCs arise from primordial germ cells (PGCs) and serve to establish, renew, and expand the germ cell population. Differentiation of PGCs to GSCs is conserved and critical for establishment of the germline, yet a major gap in our knowledge of reproductive development and fertility remains understanding what factors influence this process. To examine early GSC development, I am using zebrafish – a genetically tractable model with high fecundity and rapid external development. Recently published data from our laboratory showed that, in zebrafish, PGCs lacking the conserved germline-specific RNA-binding protein Dazl (Deleted in Azoospermia- like) fail to differentiate into GSCs and are lost shortly thereafter. Dazl is a translational regulator which was previously implicated in meiosis; however, our finding that dazl mutant PGCs fail to differentiate raises interesting questions as to how GSCs arise from PGCs, and the specific role(s) of Dazl during this process. I hypothesize that Dazl translationally regulates its associated factors during PGC differentiation to maintain PGC germline fate and promote GSC establishment. However, Dazl has been characterized as both an activator and repressor of translation of thousands of germline targets in various contexts, making the specific mechanisms by which it promotes GSC establishment unclear. Herein, I propose to functionally define Dazl’s role in GSC establishment using a combination of genetic and molecular approaches. Specifically, my proposed research strategy will (1) trace dazl mutant germ cell fates to determine whether Dazl maintains PGC identity, (2) screen for GSC-specific markers to investigate the mechanism of GSC fate establishment, and (3) test the functions of conserved Dazl- associated mRNAs which I hypothesize are translationally regulated by Dazl during PGC differentiation. Together, these approaches will delineate Dazl’s role in PGC differentiation and GSC specification. More broadly, these studies may shed light on mechanisms to maintain, renew, and prevent premature loss of the GSC population, which has important implications for reproductive health. Completion of these aims will provide rigorous training in all aspects of genetics-based functional analysis, including generation, characterization, and validation of mutant and transgenic zebrafish lines, Cre-based lineage tracing, and genomic cloning, which will foster my development as a successful independent investigator committed to using genetic models to study reproductive development.

NIH Research Projects · FY 2024 · 2023-09

The opioid epidemic remains a major public health crisis in the US, with relapse rates and overdose-related fatalities continuing to rise. However, the mechanistic explorations of viable interventions in individuals with opioid use disorder have been particularly scarce. Here we will explore a novel brain-based intervention to decrease craving in individuals with heroin use disorder (iHUD) in early treatment. A core characteristic of drug addiction is an enhanced reactivity to drug related cues and reduced processing of other reinforcers in the natural environment, as reliably observed across numerous brain networks and associated with enhanced craving (a predictor of drug use outside the lab). Our recent studies in iHUD suggest that this brain-behavior cue-induced biased pattern improves with abstinence/treatment. Therefore, we will test whether preemptively changing such neural cue reactivity could expedite the recovery process as measured with reduced drug craving. Specifically, we hypothesize that training can help iHUD achieve an intentional modulation of their cue reactivity signal. Using one’s own brain signal, real-time fMRI neurofeedback (rt-fMRI NF) allows participants to volitionally modulate brain activity in targeted brain regions shown in smokers and heavy alcohol drinkers to be effective in reducing drug cue neural reactivity, as associated with abstinence/decreases in craving. However, the permeability of this approach is not uniform. Here for the first time, we will test whether the NF effect can be enhanced by using the signal derived from the brains of others (i.e., brain-to-brain neural transmission). Specifically, our first aim in this cutting-edge exploratory application is to identify the brain regions that distinguish between early (abstinent for <1 month) as compared to later (abstinent for >3 months) time-in-treatment in iHUD. Our second aim is to use the unique multivariate neural patterns derived later in treatment as NF provided to a newly recruited sample of iHUD in early treatment, with the goal of increasing neuronal coupling between both groups. To increase ecological validity and better approximate actual real-world experiences in iHUD, the stimulus is a dynamic, narrative-based, and context-rich movie. Our working hypothesis is that, as compared to iHUD in early treatment where drug cue reactivity is more automatic and harder to control (impacting non drug processing), recovering individuals are better able to regulate it allowing them to reduce craving (and curtail, or entirely eliminate, drug- seeking) even in potent drug-related situations. Therefore, during rt-fMRI NF, we expect greater recovery (and lower cue-induced craving) in the new sample of iHUD early in treatment who show the most neural coupling/alignment with the neural patterns of those in later recovery. In short, in this innovative R21 proposal we will answer the following question: Can the neural patterns of addicted individuals in later recovery be used to provide scaffolding for the nascent recovery process early in treatment, helping to reduce craving? Results of this proof-of-concept study can be used in later longitudinal studies to develop real-time NF-based training to improve outcomes in iHUD as generalizable to other substance use disorders.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY In the midst of the opioid epidemic, methamphetamine use is emerging as the next substance use crisis with disproportionately increasing prevalence in men who have sex with men (MSM), high relapse rates and no FDA-approved treatments for individuals with methamphetamine use disorder (MUD). One of the major contributors to relapse is the enhanced motivated arousal (cue-reactivity) to drug-related cues, even after protracted abstinence that leads to an increase in craving and drug-seeking. Although cue-reactivity is studied widely in other substance use disorders, its study is scarce in MUD and non-existent in MSM with MUD, creating a critical gap in knowledge needed for unbiased assessment of disease severity and treatment outcomes in this rapidly growing clinical population. Therefore, in this bi-phased study, we propose to first identify a psychophysiological marker of methamphetamine cue-reactivity and its incubation with abstinence from methamphetamine use and examine group-differences between MSM and non-MSM MUD. Subsequently, in the second phase we propose to longitudinally assess incubation of cue-reactivity, its reduction with cognitive reappraisal (CR; a self-regulation technique) and examine the impact of CR on clinical outcomes in MSM with MUD. In light of preliminary findings from our group, we hypothesize that LPP, an electroencephalography-derived marker of drug cue-reactivity, will track the incubation of methamphetamine cue-reactivity and its CR-mediated reduction, which in turn will be associated with improved clinical outcomes in MSM with MUD. In the R61 phase, we will cross-sectionally compare the LPP-assessed cue-reactivity between currently using (Group 1) and abstinent (1-3 months; Group 2) individuals with MUD (50% MSM in each group, matched on HIV status), and examine in-task changes in cue-reactivity with CR technique. In the R33 phase, we will assign a cohort of MUD to either the CR+ (Cognitive Reappraisal training) or the CR- (no Cognitive Reappraisal training) group (all MSM, matched on HIV status), and cue-reactivity will be assessed longitudinally at <2 weeks, 2 months, to 3 months after abstinence initiation. Successful completion of these aims will identify an EEG-based, objective, and clinically useful marker of methamphetamine cue-reactivity, and will delineate the impact of CR on methamphetamine cue-reactivity in relation to treatment response in MSM with MUD. This work will be the beginning of mechanistic research into the role of incubation of methamphetamine cue-reactivity and its reduction as a reliable and clinically meaningful outcome measure, establishing an empirical foundation to develop an intervention for MUD and possibly in other addictions.