Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2026 · 2023-02

SUMMARY Acute myeloid leukemia (AML) is a clonal hematopoietic stem and progenitor cell malignancy characterized by poor clinical outcomes. Malignant transformation triggers expression of surface proteins that serve as “danger signals”, such as MICA and MICB (MICA/B) that are commonly expressed by leukemia cells in response to cellular stress pathways. Natural killer (NK) cells and cytotoxic T lymphocytes recognize MICA/B with the NKG2D receptor, which in turn induces cytotoxic functions against leukemia cells. However, they often escape recognition by shedding MICA/B via an intriguing post-translational modification called proteolytic cleavage. In previous and revolutionary study, we developed a series of MICA/B-targeted monoclonal antibodies that inhibit the shedding of MICA/B by blocking the extracellular domain that undergoes an unfolding by disulfide isomerase, which enables subsequent cleavage by metalloproteases. One of them was further characterized, and it promotes NK cell-mediated immunity against solid tumors by dual signaling of NKG2D and Fc receptors. Recently, we discovered that the antibody also promotes macrophage-driven immunity in AML models, by enabling antibody-dependent phagocytosis of leukemia cells in the blood and bone marrow. In this setting, MICA/B serve as “leukemia antigens” for Fc receptor-driven immunity, although contributions by NKG2D, NK cells, and T cells were also detected. Now we developed a new version of this antibody, with three point mutations in the Fc domain to increase the binding affinity to all three Fc activating receptors while maintaining low affinity to the Fc inhibitory receptor. This new version is humanized and induces potent NK cell effector functions against human AML cells, compared to the humanized wild type version. We also established mouse models of human Fc receptor biology and both murine and human AML models, to establish the anti-leukemia activity of our now enhanced molecule in vivo. Furthermore, we propose a mechanism-driven drug combination regimen, whereby a histone deacetylase inhibitor (romidepsin) induces human leukemia cells to express MICA/B mRNAs that are then translated to MICA/B proteins, which are followed by stabilization on cellular surface by our antibody that inhibits the shedding. We propose that immunotherapy for AML can be achieved via this drug combination, which increases the innate immunogenicity of leukemia cells. For these reasons, our studies will generate important information about how to induce protective immunity against AML. Antibody-mediated inhibition of MICA/B shedding is a new immunotherapeutic opportunity pioneered by us and now validated independently by multiple pharmaceutical companies, one of which has recently began phase-I clinical trial testing an antibody analog to ours, but the wild type version, in patients with advanced solid tumors. Therefore, our studies will generate compelling rationale to test MICA/B-targeted antibodies in AML trials, by pharmaceutical companies, ourselves in academia, or both by working in collaboration.

NIH Research Projects · FY 2026 · 2023-02

Project Summary/Abstract The significance of sleep and circadian disorders on public health is broad because of its effects on physical and mental health. The high prevalence of these disorders makes it critical that a robust pipeline exists for new investigators to tackle the challenge of understanding the mechanisms involved, their impact on cardiovascular, metabolic, pulmonary and neurodegenerative systems and develop and test interventions. Faculty at Icahn School of Medicine (ISMMS) and New York University Grossman School of Medicine (NYU) from multiple departments (sleep medicine, pulmonary, psychiatry, neuroscience, radiology, population health, cardiology, and biostatistics) have been involved in collaborative basic and clinical research in sleep and circadian science for several decades. Our training program will build on the foundation of successful mentored research and training programs (e.g. PRIDE institute at NYU) with experienced, well- funded investigators who have demonstrated a strong commitment for career development. The areas of current research include (but are not limited to) (i) basic biology of the circadian clock, (ii) translational research on the role of sleep on memory, (iii) clinical patient oriented research evaluating physiology of obstructive sleep apnea (OSA), its treatment and evaluation of outcomes of OSA on cardiovascular and brain health (iv) developing simulation and web based tools to assist teaching sleep physiology. State of the art physiological and imaging techniques (cardiac CT imaging, PET/MR brain imaging, Alzheimer’s disease biofluid biomarkers) are employed, and include subjects across the age spectrum (pediatric to elderly populations) and minority populations. We will leverage access to the existing training infrastructure at the two institutions along with the experience of successful collaborations over the past 10 years between multidisciplinary faculty at the two institutions and incorporate translational content to train post-doctoral fellows. The aim of this 5-year program is to recruit and train a total of 6 postdoctoral candidates (physician scientists or PhD graduates). Trainees will participate in a 2-3 year program during which time they are assigned to a mentoring team, comprised of a primary, secondary and peer mentor. Mentors are nationally recognized investigators with expertise in guiding mentees in developing their careers and in submitting successful grant applications. Trainees will implement an Individual Development Plan, use a combination of existing conferences, didactic courses and technology- assisted multi-institutional workshops to acquire professional, laboratory based skills and training in responsible conduct of research. Trainees will have access to the broad training and research support resources of the two institutions including those of the CTSI/CTSA. Measurable outcomes will include trainee publications, presentations, and grant applications. Mentees’ achievements for 10 years post program will also be evaluated and include academic leadership positions, career awards, federal and non- federal grants and engagement in mentoring activities. The Sleep Science T32 Program addresses the need for sleep and circadian researchers and facilitates their transition to independence.

NIH Research Projects · FY 2026 · 2023-02

Tandem Repeat Expansions (TREs), most commonly of triplet repeats such as poly(CAG), are known to underlie >40 different human neurological diseases. While the majority of TREs identified to date have been found in late-onset neuro-degenerative disorders such as hereditary ataxias and Huntington disease, TREs have been identified in patients with AD and certain types of dementia. In addition to these expansions of short tandem repeats (STRs, with motif sizes between 1 and 6 base pairs), copy number variation of larger repeats with motifs ≥10bp, also known as Variable Number of Tandem Repeats (VNTRs), has recently been linked to risk of AD. However, despite this evidence that variation in tandem repeat (TR) sequences can act as the causative mutations in some cases of AD and dementia, there have been no concerted efforts in AD cohorts to either systematically screen for novel TREs, or to genotype VNTR copy numbers. Newly developed bioinformatic approaches that can be applied to analyze Whole Genome Sequencing (WGS) data now provide an opportunity to fill this knowledge gap. Utilizing the expertise and knowledge that we have gained working on other large datasets, we propose to apply these approaches to analyze ~62,000 genomes sequenced by the Alzheimer's Disease Sequencing Project that are available to the community, in addition to a further ~48,000 control genomes from TOPMed, and will use these data to investigate two major hypotheses: 1. We hypothesize that some cases of AD are caused by rare, highly penetrant pathogenic TREs. Using novel bioinformatic tools that can identify TREs, we will search for rare TREs that are observed only in AD samples, or which show significant enrichment in AD cases compared to controls, and thus are likely causative for AD. Potentially pathogenic TREs will then be validated by PCR or long-read sequencing in available DNA samples. 2. We hypothesize that common polymorphic copy number variation of TRs of all motif sizes can act as genetic risk factors for AD. Short tandem repeats (STRs) with motif sizes 1-6bp will be genotyped with hipSTR. VNTRs will be genotyped using read depth to estimate diploid copy number. We will analyze available WGS data from ~20,000 sporadic late-onset AD samples and ~90,000 unrelated controls, generating copy number estimates for >1 million STRs and ~150,000 VNTRs genome-wide, which will be used to perform association analysis of TR copy number with AD status in a case:control study. Given that TREs, and polymorphic variation in STRs and VNTRs, both represent established mutational mechanisms that contribute to a variety of late-onset neuro-degenerative conditions, we believe that the study of TRs in AD represents a logical step that has a high likelihood of uncovering novel genetic causes of AD.

NIH Research Projects · FY 2026 · 2023-02

Project Summary/Abstract Low weight eating disorders, such as anorexia nervosa/atypical anorexia nervosa, are severe psychiatric disorders that often develop in adolescence, have a chronic course, and evidence poor response to treatment at great public health cost. Two core features of LW-EDs are food avoidance and preoccupation with shape and weight. Preliminary data from our initial mechanism study indicate that family based interoceptive exposure (FBT-E) therapy reduces food avoidance, increases autonomous eating and nonjudgement body awareness, and enhances extinction learning to aversive food cues to a greater degree than existing family- based therapy (FBT). Feasibility data indicated high retention, treatment fidelity, and established reliable measures of treatment adherence despite similar short-term (6-week) within-treatment changes in expected body weight. Long-term (6-month) follow-up of these interventions in open trial indicate significantly higher expected body weight among those having received FBT-E. Short-term and long-term changes in weight were highly correlated with changes in expected body weight. Consequently, we will test the confirmatory efficacy of FBT-E vs FBT and 12-month follow-up in a parallel group randomized clinical controlled trial in 120 adolescents aged 12-18. We have three specific aims, including: (1) to determine short term and long term efficacy of FBT-E vs FBT in expected body weight, global eating disorder symptoms, and clinical impairment; (2) to determine efficacy of FBT-E vs. FBT on laboratory measures of autonomous eating, nonjudgmental body awareness, and extinction learning; and (3) estimate whether temporal precedence and mechanistic value (mediation) of early changes in autonomous eating, non-judgmental body awareness, and extinction learning on treatment effects and longitudinal outcomes. Assessments and data analyses will be blinded to treatment. With N=120 participants (n = 60 per group) we can detect moderate sized effects for primary and secondary outcomes. Results will provide data to support novel treatment for LW-EDs and provide theoretical robust model for understanding pathology for children and families that suffer with LW-EDs.

NIH Research Projects · FY 2026 · 2023-02

Bladder cancer is the most expensive cancer per capita to treat in the US. Non-muscle invasive bladder cancer (NMIBC) which accounts for 70-75% of all newly diagnosed tumors and has only a single FDA approved first- line treatment option, Bacillus Calmette-Guérin (BCG). BCG has been the only approved first-line therapy for intermediate and high-risk NMIBC for more than 40 years. While BCG can induce durable responses, ~50% of patients have recurrence or progression of their disease. The current dosing of BCG is arbitrary with no data currently available to support the dosage, strain nor regimen. Furthermore, no clear mechanism of action behind BCG’s anti-neoplastic activity in NMIBC has been delineated. Better understanding of the immune response initiated by exposure to BCG and how such response is sustained over many repeated doses is essential to help improve its therapeutic action. Further, it may lead to discovery of predictive biomarkers and novel targets leading to advance care for patients, reduce the financial burden associated with the disease, and improve access to BCG. Our preliminary data have identified a significant Type-1 cytokine response to BCG in all patients inducing an activation and recruitment of IFN-g producing NKG2A+ natural killer (NK) cells and NKG2A+PD-1+ CD8 T cells to the tumor microenvironment (TME). Our preliminary analyses of post-BCG-treated specimens demonstrate IFN-g signaling as the most upregulated signature in BCG resistant specimens and strongly correlates with increased HLA-E and PD-L1 expression on the recurring tumors. Monalizumab (anti-NKG2A) is a novel therapeutic target and is currently in clinical trials across several cancer types. A role for NKG2A/HLA-E alone or in conjunction with PD-1/PD-L1 in regulating NK and CD8 T cells remains elusive with virtually nothing known in bladder cancer. Our hypothesis is that chronic activation from repeated exposures to BCG drives immune dysregulation followed by functional exhaustion in a significant proportion of patients, which then leads to disease recurrence or progression. The immune dysregulation in the TME is driven by NKG2A/PD-1 on CD8 T cells, NKG2A on NK cells and HLA-E and PD-L1 on tumor cells. To further validate our findings and better understand BCG mechanisms we will study the following aims: Aim 1: To analyze intratumoral NK cells and T cells and their interactions with tumors in response to BCG therapy. We will test the hypothesis that upregulated expression of HLA-E and PD-L1 on recurring tumors are directly in contact with infiltrating NKG2A+ NK and NKG2A+ PD-1+ CD8 T cells. Aim 2: To longitudinally assess blood and tumor phenotypes and functions in response to BCG therapy. We will test the hypothesis that repeated exposures to BCG drives prolonged activation and dysregulation of NKG2A+ NK and NKG2A+ PD-1+ CD8 T cells. Aim 3: To determine the effects of combination PD-L1 and NKG2A blockade on anti-tumor immunity. We will test the hypothesis that NKG2A+ NK and NKG2A+ PD-1+ CD8 T cells can be functionally restored through use of PD-L1 and NKG2A combination blockade.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY/ABSTRACT Recapitulation of normal lung function following a severe acute injury implies an inherent regenerative ability of the lung. However, sources and relative regenerative capacities of lung epithelial stem/progenitor cells remain unclear, especially in the human lung. Depending on the injury type and severity, several distinct progenitors are activated and respond by proliferating and differentiating to aid in near complete recovery. Both airway and alveolar stem/progenitor cells are activated and contribute to alveolar repair following severe injuries such as influenza or bleomycin. To this end, recent studies from our lab have uncovered an airway epithelial progenitor cell marked by elevated levels of Major Histocompatibility Complex (MHC) Class I protein, H2-K1. Despite having a transcriptome highly similar to the mature club cells, the H2-K1high progenitors, unlike mature club cells, selectively proliferate post injury and aid in improved oxygenation in injured mice after orthotopic transplantation. However, mechanisms underlying early and selective activation of these progenitors remain unknown. In addition, there are several more aspects of distal lung regeneration that are yet unclear. The chief among them is whether the distal epithelial progenitor hierarchy that we observe in mouse lungs is maintained in distal human lungs. The distal human lung airways have a higher proportion of basal cells and have more heterogeneous secretory cell populations than the mouse airway epithelium. Furthermore, the human type 2 alveolar epithelial cells (AEC2s) have remarkable in vitro proliferative and regenerative capacity. Therefore, there is an unmet need to understand the identity and regulation of distal human airway and alveolar progenitors. To this end, our preliminary data show that a distal human secretory subpopulation that is analogous to the mouse H2-K1high club-like progenitors can give rise to AEC2s in vitro. Conversely, we have uncovered a novel and unexpected ability of mature human AEC2s to differentiate towards airway lineages in vitro and in vivo, suggesting at least two sources of distal epithelial regeneration in the human. Therefore, it is critical to clarify the identity and characteristics of both distal mouse and human epithelial progenitors primed for alveolar repair. This proposal seeks to answer these questions through three aims: 1) Determine the mechanisms underlying activation of H2-K1high progenitors in alveolar repair. 2) To identify distal secretory cells as a source of alveolar cells post injury in the distal human lung. 3) Determine whether a subpopulation of mature human AEC2s has reversible bi-directional potential to differentiate into alveolar basal cells. These aims will utilize single cell mRNA and open chromatin sequencing, in vitro and in vivo manipulation of proposed signaling pathways, and orthotopic transplantation of progenitor cells to clarify the epithelial stem/progenitor cell hierarchy in distal mouse and human lungs. These studies will lay the foundation for interrogating several novel signaling pathways to ultimately aid in our efforts to manipulate regenerative mechanisms to achieve better disease outcomes.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Disrupted communication between brain regions responsible for emotion regulation (limbic and higher cognitive cortical regions) may critically underlie the emotional dysregulation that characterizes mood and anxiety disorders. However, the most instructive metric of such communication has yet to be agreed upon. In addition, due to technical barriers, there are currently no fine-grained measurements of limbic subregion connectivity in humans. A network- based approach can be used to explore limbic and cortical subregion connectivity (i.e. connectome), shedding light on the discrete or shared neural mechanisms underlying core symptoms in mood and anxiety disorders. In the proposed study we will (1) characterize small limbic subregions and whole-brain connectomes in healthy controls and individuals with major depressive and generalized anxiety disorder, by optimizing and employing a novel 7-Tesla MRI protocol with improved spatial resolution with whole brain coverage; (2) discern network based biomarkers of depression and anxiety by developing and characterizing a multi-modal integrative brain network consisting of structural, functional and dynamic topographies using a new computational multilayer approach; and (3) transdiagnostically examine how connectome organization manifests across the three study groups. By portraying specific limbic subregion involvement in the brain connectome, this would advance our understanding of how connectome alternations manifest in psychiatric disorders. Furthermore, this work combines high-resolution measures of brain activity and synchronization (functional MRI), with maps of the brain’s white matter architecture and anatomical connections (diffusion MRI). Their combined study will facilitate identification of aberrant network features and their contributions to core symptoms in depression and anxiety. Lastly, the transdiagnostic approach will uncover shared and unique network mechanisms of depression and anxiety that could potentially provide improved diagnostics and treatment selection. This Career Development Award will allow for the critical complimentary training goals centering on: (1) gaining practical clinical experience towards identifying clinical needs and conducting translational research; (2) development of technical advanced MRI sequence development skills; (3) refine computational skills in advanced network science methods. The proposed research and training afforded by this award will allow me to launch an independent research program developing neuroimaging methods to study brain network perturbations in mood and anxiety disorders.

NIH Research Projects · FY 2025 · 2023-01

Project Summary Synucleinopathies, including dementia with Lewy bodies (DLB) and Parkinson’s disease (PD), are a growing health crisis, affecting over 1 million people in the United States and an estimated 10 million people globally. Synucleinopathies are caused by accumulation of the protein α-synuclein (α-syn) into unnatural fibrils in the brain and can manifest with motor symptoms, cognitive symptoms, or a combination of the two. There are currently no effective treatments to cure or prevent synucleinopathies, in large part because there is no clear cause for disease. There are many identified genetic and environmental risk factors associated with synucleinopathies, but the context in which these factors lead to disease is still unknown. In addition to direct mutations to SNCA, the gene that encodes α-syn, mutations to GBA and APOE are well established risk factors and have been shown to increase α-syn aggregation. However, none of these risk factors are great predictors of disease on their own, suggesting unknown genetic and environmental interactions likely influence the initiation, severity, and clinical outcomes of α-syn pathology. In this proposal, I focus on how both cell type and genotype interact with each other to modulate α-syn pathology severity. I hypothesize that α-syn pathology is precipitated by epistatic interactions between genetic factors that disrupt homeostasis in different cell types and leaves dopaminergic neurons vulnerable to deleterious α-syn pathology and neurodegeneration. I will investigate this hypothesis using two independent, complementary approaches to dissect the cellular and molecular mechanisms that modulate α-syn phenotypes. First, I will couple GBA knockdown with SNCA-triplication, wild-type, and knockout cell lines. There is a clear link between GBA mutations and α-syn aggregation, but how these proteins connect is still unknown. I will use high-content imaging with genetically encoded fluorescent markers to quantify lysosomal and mitochondrial dynamics in live cells. This will reveal how GBA responds to different α-syn dosages and how that affects cellular homeostasis. Second, I will leverage isogenic lines containing APOE allelic variants and GBA knockdown. GBA and APOE are involved in lipid metabolism, suggesting a putative link between lipid metabolism disruption and α-syn pathology, although there is no published connection between the two proteins. I will use next generation genomic sequencing and proteomic approaches to determine the interacting effects between SNCA, GBA, and APOE allelic variants on α-syn pathology severity. Completion of this aim will uncover whether GBA and APOE are acting through converging or independent pathways, expanding our knowledge of the network that contributes to disease pathology. By coupling multiple genetic risk factors into a multi-cell type model, I will pioneer new technology and approaches to dissect epistatic mechanisms that influence α-syn pathology. These insights will guide us in developing new therapeutic approaches, enabling earlier detection, treatment, and prevention of synucleinopathies.

NIH Research Projects · FY 2026 · 2023-01

The overall goal of this proposal is to identify optimal strategies for primary prevention of cardiovascular disease (CVD) for survivors of breast (BC), prostate (PC) and lung (LC) cancer. All together, there are >6 million BC, PC and LC survivors in the US. While cancer is a major source of morbidity and mortality, the majority of BC and PC as well as many early-stage LC survivors die of comorbidities, particularly CVD. Cancer survivors have increased rates of both CVD risk factors as well as CVD itself, and CVD is the leading cause of death among BC and PC survivors. Among LC survivors, CVD-related deaths account for ~30% of mortality; this proportion is higher among the growing number identified with early-stage LC. In the general population, primary prevention with lipid-lowering agents (i.e., statins) and aspirin is highly effective for decreasing CVD incidence and mortality, but these guidelines for primary CVD prevention cannot be extrapolated to cancer survivors. Approaches for CVD prevention in cancer survivors need to consider traditional risk factors (including genetic risks) for CVD as well as CVD risk from certain cancer treatments. Additionally, competing risks from cancer recurrence or comorbidities may limit the long-term benefits of primary CVD prevention. Finally, the cancer itself, cancer treatment-related complications, and a higher prevalence of comorbidities can negatively impact quality-of-life and attenuate the absolute improvement in quality-adjusted life expectancy and the cost effectiveness (CE) of primary CVD prevention. Lack of specific data applicable to cancer survivors has profound negative impact, resulting in worse cardiovascular outcomes. It is unlikely that randomized controlled trials (RCT) assessing the benefits of CVD preventive strategies for cancer survivors will be ever conducted. Thus, there is an urgent need to use alternative methods to optimize preventive care recommendations for this growing population. We propose using simulation modeling, an approach complementary to clinical trials, to assess the harms, benefits, and CE of CVD prevention in a representative population of cancer survivors.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY All new drug candidates must be tested for their potential to cause arrhythmia as a drug-induced adverse event. Recent years have seen substantial progress in developing more sensitive and specific predictions of which drugs may increase arrhythmia risk, and my group has been at the forefront of efforts to employ mechanistic modeling for quantitative predictions of cardiac drug safety. Nonetheless, arrhythmias are rare events, and even drugs that are considered dangerous only induce arrhythmias in a minority of patients. Therefore, identifying which patients are most at risk of arrhythmia, and which conditions increase their risk, is as important as classifying drugs. The work proposed here will address these challenging questions through an innovative combination of: (1) in vitro physiology experiments that will quantify how drugs influence myocyte action potentials and intracellular calcium; (2) simulations with mechanistic mathematical models that incorporate phenotypic differences between groups and between individuals within the same group; and (3) machine learning to synthesize results and develop predictive classification systems. Experiments performed in stem cell-derived myocytes will measure cellular responses to a wide range of drugs, and these data will allow for rigorous tuning of mathematical models. Subsequent simulations of heterogeneous populations will address challenging unresolved questions, such as: 1. How do patient characteristics influence arrhythmia risk? Simulations will address how sex differences in cardiac electrophysiology and the presence of pre-existing cardiac disease influence drug responses. Through a combination of cellular experiments, mechanistic mathematical modeling of heart cells, and machine learning models, we will quantify how much each factor influences arrhythmia risk. 2. How do symptoms associated with common diseases influence the potential pro-arrhythmic effects of drugs used to treat those diseases? Many diseases are associated with conditions that influence cardiac electrophysiology, such as fever, hypokalemia, and chronic inflammation. We will develop a simulation platform that accounts for these effects. 3. Which patients within a group are especially at risk? Besides quantifying the effects of differences between groups, our machine learning classifiers will allow us to predict which patients within a group are especially susceptible to drug-induced arrhythmia on the basis of their “electrophysiological signatures.” Together these studies will offer a new paradigm for quantitative understanding and prediction of drug-induced arrhythmia that considers not only differences between drugs, but also between the patients that take these drugs.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY: This is a revised version of a new R01 grant that characterizes stable epigenetic changes that are induced in mouse limbic brain regions in response to early life stress (ELS) which then increase an individual's susceptibility to subsequent stress for a lifetime. This work has been supported over the past ten years by an NIMH Conte Center, however, that Center has now “sun-setted”—it cannot be renewed—hence, this new proposal to continue this original line of research. Our innovative hypothesis is that ELS induces stable “chromatin scars” that drive long-lived changes in gene expression, which then mediate downstream changes in cell and circuit function and ultimately behavioral responses to chronic stress in adulthood. We concentrate on nucleus accumbens (NAc) based on its central role in brain reward and motivation and on empirical findings of its central role in controlling stress susceptibility. We utilize open-ended, unbiased proteomic and next- generation sequencing approaches to first identify the most robust and significant putative chromatin scars in NAc in a cell-type-specific manner and to then characterize those mechanisms at the molecular, cellular, and behavioral levels. Importantly, we investigate only those mechanisms that are validated in postmortem NAc of depressed humans. One major focus is H3K79me2 (dimethylation of Lys79 of histone H3), the induction of which is the most significant histone modification caused by ELS in NAc of both male and female mice. The methyltransferase (DOT1L) and demethylase (KDM2B) that control H3K79me2 are both induced in NAc by ELS, effects specific to D2-type medium spiny neurons (MSNs). Bidirectional viral-mediated manipulation of DOT1L or KDM2B selectively in D2 NAc MSNs of male and female mice establishes the role played by these enzymes in mediating the ability of ELS to increase stress susceptibility based on rapid behavioral screening assays. We will now extend measures of stress susceptibility to numerous, more sophisticated behavioral procedures with greater translational potential, and test the ability of a DOT1L inhibitor, now in clinical trials for certain cancers, to reverse ELS-induced stress susceptibility upon systemic administration, further promoting the translational potential of the proposed research. As well, we have shown by RNAseq that DOT1L overexpression in D2 MSNs mimics a large portion of gene expression changes induced by ELS in this cell type, while DOT1L knockdown blocks ELS action. We will now map H3K79me2 enrichment genome-wide in D2 MSNs by CUT&RUNseq to identify networks of genes whose altered expression by ELS is mediated by this histone mark. We will also generate more complete open-ended proteomic and transcriptomic datasets and characterize additional putative chromatin scars induced in NAc by ELS. Substantial preliminary data, for example, implicate H3K27me1 as a prominent chromatin scare that predominates in D1 MSNs of male mice only. Together, this work will characterize novel mechanisms that drive a persisting state of enhanced stress susceptibility and offer insight into new ways of reversing such susceptibility in adulthood.

NIH Research Projects · FY 2026 · 2022-12

The preferred vascular access for hemodialysis to treat end-stage renal disease involves using a vein as a conduit to increase blood flow by surgically creating an arteriovenous fistula (AVF). Successful adaptation of a venous conduit to the fistula or arterial environment requires remodeling of the vein wall without excessive wall thickening, enabling mechanical strength to resist hemodialysis procedures that puncture the AVF wall with large bore needles 3 times a week. However, the poor maturation and patency of AVF, especially in women and requiring additional re-do procedures and surgery, reflects our imperfect understanding of the biology of venous remodeling that leads to successful venous adaptation to the fistula environment. This knowledge gap creates an unmet need for novel approaches to enhance venous remodeling and thereby to increase successful clinical use of venous conduits. Using a mouse AVF model that recapitulates human AVF maturation and shows sex differences, we have shown that both an innate immune response as well as an adaptive immune response regulate venous remodeling. We present exciting new data that sex hormones mediate sex differences in wall thickness during venous remodeling. In addition, we have developed the mouse model further to incorporate chronic kidney disease (CKD) via 5/6-nephrectomy; AVF in the CKD environment show altered venous remodeling compared with control mice and these AVF faithfully recapitulate human AVF maturation. We hypothesize that since T cells mediate venous remodeling, modulating adaptive immunity will alter venous remodeling, thereby improving AVF patency and ultimately fistula utilization in human patients. We will use our translationally relevant in vivo model, an innovative tool encapsulating cyclosporine in nanoparticles for local drug delivery, innovative methodology to analyze the immune cell composition within the AVF wall, as well as advanced next- generation analyses using transcriptomics techniques that are available at Yale, to test our innovative hypothesis with the following specific aims: Aim I: Determine whether sex differences in adaptive immunity between women and men affect AVF remodeling in vivo. Aim II: Determine whether sex hormones mediate sex differences in the immune response during AVF remodeling in mice with CKD. Aim III: Determine whether PD-L1 expression regulates the effects of adaptive immunity on AVF remodeling in mice with CKD. A successful outcome of this investigation will have lasting impact by establishing whether there is a T cell basis underlying venous remodeling and thus manipulation of adaptive immunity is a valuable strategy for clinical translation to enhance AVF patency. We will also determine whether reduced AVF maturation in women is due to sex differences in adaptive immunity. We use an innovative strategy and novel tools and models to manipulate adaptive immunity to alter venous remodeling and thereby improve AVF patency.

NIH Research Projects · FY 2026 · 2022-12

After the 9/11/2001 terrorist attack on the World Trade Center (WTC), >100,000 residents, police, firemen, emergency medical technicians, and others were chronically exposed to an environmental toxic cloud of dust and chemicals for more than a year during the massive cleanup effort. Adequate respiratory protective equipment was not consistently available, so these exposures contributed to health consequences presenting years later. Understanding the impact of 9/11 on long-term cardiometabolic disease (CMD) risk is critical because federally funded healthcare is only provided to responders for diseases certified to be related to 9/11 exposure. Current evidence of WTC health effects is mostly limited to respiratory and cancer risk, while myocardial infarction (MI) and diabetes are not certified. This has vast implications for health care costs and accessibility of treatment. We propose a retrospective cohort study using the WTC Health Program (WTCHP) General Responder Cohort to identify WTC-related CMD risk. Mount Sinai is home to the WTCHP Data Center, a repository of all monitoring and treatment data from all five Clinical Centers of Excellence that longitudinally monitor responders involved in the 9/11 tragedy. The WTCHP has assessed more than 43,000 responders during 20 years of follow-up with physical examinations, laboratory tests, and exposure and health questionnaires. With this cohort we will innovatively address gaps in WTC-related CMD research; assess the effect of the 9/11 attack on diabetes and MI incidence, as well as glucose and total, HDL, and LDL cholesterol levels; and identify how this exposure interacts with subsequent environmental exposures. In Aim 1, we will estimate the association between WTC- related exposures and CMD. In Aim 2 we will assess the association between long-term exposure to fine particulate air pollution (PM2.5) chemical components and CMD. We have developed a novel spatiotemporal model that can identify PM2.5 components at very high spatial resolution, allowing us to identify mixtures that define effects. Unlike most current research, we will address the complexity of PM2.5 exposure as a mixture of chemical components rather than focusing on PM2.5 mass. In Aim 3, we will determine whether subsequent environmental exposures (i.e., air pollution, greenness, noise, walkability, food environment, social vulnerability, and temperature) interact with the effects of WTC-related exposures on CMD. We will assess whether WTC-related exposures among responders altered their susceptibility to subsequent air pollution exposure, setting up distinct cardiometabolic health trajectories. We will also identify beneficial environmental exposures that mitigate the cardiometabolic effect of WTC-related exposures. To our knowledge, this will be the first study to examine whether subsequent environmental exposures interact with WTC-related exposure effects on CMD. This research will leverage a rich cohort with 20 years of follow-up data to uncover the long- term trajectories of how environmental exposures contribute to CMD, revealing critical insights to inform future interventions and policies to protect human health from hazardous exposures.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY A vaccine is a promising approach for stopping the spread of HIV infections. Although vaccine regimens in clinical trials show various levels of protection against HIV, there is no effective vaccine available for a large population yet. Preclinical and clinical data demonstrate the importance of both adjuvants and antigens for an effective HIV vaccine. Particularly, stimulation of toll-like receptors (TLRs) or stimulator of interferon genes (STING) boosts the immune response of the HIV antigens in multiple animal models. Meanwhile, HIV immunogens are a critical factor for both humoral immunity and cell-mediated immunity such as broadly neutralizing antibodies (bnAbs) and cytotoxic T cells. Despite these important advances, significant challenges remain in immunogen design, immunogen delivery, and adjuvant choice. To overcome these challenges, we propose to integrate adjuvant derived nanoparticles and engineered mRNA for HIV vaccine discovery. In preliminary studies, we developed adjuvant derived nanoparticles (ANPs) using TLR or STING agonists, which showed great potential for efficient mRNA delivery as a vaccine platform. Moreover, we constructed glycosylated HIV immunogens that trigger mannose-binding lectin (MBL)-mediated innate immune recognition, leading to enhanced antibody responses. Additionally, we systematically investigated the untranslated regions (UTRs) of mRNAs in order to enhance protein production. Through a comprehensive analysis of endogenous gene expression and de novo design of UTRs, we identified the optimal combination of 5’ and 3’ UTR for mRNA engineering. Based on these results and findings, the goal of this proposed project is to develop adjuvant derived nanoparticles as functional nanomaterials capable of efficiently delivering HIV immunogens in vivo, consequently generating strong and durable humoral and cell-mediated immunity against HIV. The following specific aims will be carried out to accomplish our goal: 1) Synthesis and characterization of adjuvant derived nanoparticles (ANPs); 2) Engineering of mRNA transcripts encoding various HIV immunogens with high translation efficiency; and 3) Determination of immunogenicity and safety profiles of ANPs-mRNA in mouse and non-human primate models. Encouraged by results from our preliminary studies, we expect the newly designed nanomaterials from this proposal to establish a vaccine candidate, which can facilitate clinical translation and a new avenue for HIV vaccine discovery. Knowledge gained from this study can also be extended to other types of vaccines for emerging pathogens.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY Food allergy (FA) and atopic dermatitis (AD) are common chronic conditions of atopy affecting 10% and 20% of children, respectively. Individuals with FA are at daily risk for potentially life-threatening hives, angioedema, respiratory difficulty, cardiovascular compromise, gastrointestinal distress, and/or anaphylaxis following ingestion of a food antigen to which they are sensitized. Individuals with AD must live with chronically pruritic, inflamed skin that can cover a significant proportion of their bodies and become infected. The development of FA is frequently preceded by AD, suggesting shared risk factors and overlapping pathobiology. FA and AD are complex diseases, and parsing the biological heterogeneity hidden under their clinical umbrellas is paramount to improving their prevention, diagnosis, and clinical management. A systems biology approach where the biology of FA and AD are investigated comprehensively at several levels may help identify new knowledge about the development of allergy. Systems Biology of Early Atopy (SunBEAm) is a general population pre-birth cohort study initiated by the NIAID to study the role and inter-relationships of genetic, clinical, biological, and environmental early-life factors in the development of allergic diseases, with an emphasis on FA and AD. Pregnant women, children born to them, and the children's biological fathers are being enrolled and phenotyped with longitudinal biosample collections. A central goal of SunBEAm is to apply systems biology to identify mechanisms and biomarkers underlying the development of FA, AD, and their endotypes. The overarching goal of this SunBEAm Analysis & Bioinformatics Center (ABC) application is to create a center for assaying SunBEAm biosamples using omics, and to apply integrative systems biology to identify novel determinants of FA and AD. Our application centers on these five specific aims: (1) Assay biosamples from SunBEAm participants to generate longitudinal, multi-scale omic data that will inform on the development of FA and AD; (2) Analyze the systems-wide omic data generated to identify novel biomarkers, endotypes, and mechanisms underlying the development of FA and AD; (3) Integrate data across molecular dimensions to identify key drivers and predictors of FA and AD development; (4) Share all generated data, create a centralized platform for SunBEAm data and bioinformatic innovations, and build capacity for the integration of future SunBEAm omics data; and (5) Develop infrastructure for interactions with the SunBEAm Steering Group, the NIAID Statistical and Clinical Coordinating Center, and NIAID. With this award, we will create a center for unprecedented systems allergy research. In addition to an investigator team with world- class expertise in multi-omics, systems biology, computational biology, data science, and bioinformatics, our team also includes physician-scientists and immunologists who directly designed and continue to actively drive the ongoing enrollment and implementation of SunBEAm. Our team's experience, expertise, and proven track record will ensure the success of SunBEAm-ABC.

NIH Research Projects · FY 2025 · 2022-12

PROJECT SUMMARY Tauopathies are a group of heterogeneous neurodegenerative diseases characterized by pathological aggregation of tau protein in the brain. The most common primary tauopathy, progressive supranuclear palsy (PSP), is identified clinically by progressive parkinsonism with supranuclear gaze paralysis, cognitive dysfunction, behavioral changes, and other neurological deficits. Neuropathologically, PSP has a constellation of features including morphologically distinct tufted astrocytes (TA). Astrocytes normally do not contain significant levels of tau suggesting that TA play an important, but unknown, pathological role in PSP. The majority of PSP cases are sporadic. Genome wide association studies have identified common variants associated with PSP risk in EIF2AK3, which encodes an endoplasmic reticulum (ER) stress sensor critical for the unfolded protein response (UPR). There is evidence for a pathogenic association between ER stress in astrocytes and tau accumulation in PSP, but to test candidate molecular mechanisms there is a critical need for a model system that recapitulates the cellular environment of the human brain and retains the genetic complexity of human disease. Developing human induced pluripotent stem cell (iPSC)-derived models generated from PSP patients will be a powerful tool to characterize molecular drivers of sporadic tauopathy. We hypothesize that UPR activation in astrocytes drives neurodegeneration and plays a key role in sporadic PSP. In our first aim, we will correlate UPR activation with astrocyte pathology and degeneration in human post-mortem brain tissue. Autopsy-confirmed PSP brain tissue from our large collection will be stained, imaged, and analyzed for p-tau distribution and UPR colocalization using innovative quantitative approaches. In the second aim, we will test the hypothesis that PSP neurons and astrocytes are highly vulnerable to tau pathology, and that UPR activation in PSP astrocytes increases cell vulnerability. To accomplish this aim, we will create patient iPSC-derived monolayer neurons and astrocytes and directly measure molecular changes and cell vulnerability by pharmacological manipulation of the UPR. Lastly, we will determine the extent to which UPR activation in astrocytes increases the production of toxic tau species and cell vulnerability in an iPSC-derived organoid model of sporadic PSP. This aim will explore molecular perturbations in different cell populations that contribute to the development of tau pathology in a 3-D cellular environment that recapitulates the complex neuronal-astrocyte interactions that occur in human brain tissue. These findings will be validated in paired autopsy brain tissue from the same donor. In summary, the clinical-translational research performed in this study will generate patient- derived iPSC models that retain a disease-relevant genomic background and reveal candidate pathways that are disrupted early in sporadic PSP.

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract A gap exists in our understanding of the pathophysiology behind Alzheimer’s Disease (AD), which has led to virtually nonexistent treatment options. Recent studies have identified microglia, the innate immune cells of the brain, as key players in the response to AD that may help us fill this gap. Specifically, microglia appear to play a protective role against toxicity associated with amyloid-ß containing plaques. Through activation of the triggering receptor expressed in myeloid cells 2 (TREM2) signaling pathway, microglia migrate towards and surround these plaques while inducing a distinct transcriptional signature known as the Disease Associated Microglia (DAM) phenotype. An important outstanding question is how microglia regulate this state, both transcriptionally and functionally. Our lab has previously identified the polycomb repressive complex 2 (PRC2) as an important epigenetic regulator of brain region specific microglia subpopulations. PRC2 is an epigenetic complex involved in gene silencing and has also been implicated as a signaling regulator in immune cells. Notably, we found that PRC2 deficient microglia downregulate many genes in the TREM2 signaling pathway. I hypothesize that PRC2 controls the microglial response to amyloid-ß containing plaques in a TREM2- dependent mechanism. In support of this, I generated a PRC2-deficient microglia mouse line crossed to the 5xFAD amyloid model of AD and showed that PRC2-deficiency leads to loss of plaque associated microglia, similar to TREM2-deficient models. This could be due to transcriptional regulation of the TREM2 pathway or direct modulation of TREM2 signaling. To further investigate my hypothesis, I will first characterize the epigenetic and transcriptional role PRC2 plays in 5xFAD microglia. There are multiple TREM2-dependent mechanisms that could lead to decreased plaque associated microglia – inability to sense plaques, increased death at the plaques, or lack of migration towards the plaques. To determine if PRC2 regulates these phenotypes in a TREM2-dependent manner, I will culture primary mouse microglia and pharmacologically inhibit PRC2, followed by induction of TREM2 signaling. With this model, I will assay TREM2-dependent sensing, survival, and migration. The results of this proposal will reveal whether PRC2 is a master regulator of the microglial response to AD amyloid-ß containing plaques through its control of TREM2 signaling, either transcriptionally or through direct regulation of the signaling pathway. These data are critical to furthering our understanding of how microglial functional states are regulated, opening avenues for the development of effective treatments against Alzheimer’s Disease.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY/ABSTRACT Background: Breast cancer is the most common cause of cancer incidence and mortality in women in Nigeria. Survival is poor, in part due to inadequate access to effective treatment. The National Comprehensive Cancer Network (NCCN) Harmonized Guidelines for sub-Saharan Africa (SSA) were developed to promote delivery of evidence-based, context-appropriate breast cancer care. Dr. Romanoff’s prior work shows that receiving multi- disciplinary, guideline-concordant breast cancer care in Nigeria is associated with improved outcomes. Nonetheless, guideline implementation and adherence are sporadic and have not been systematically evaluated. The overall goal of this K08 proposal is to facilitate Dr. Romanoff’s development into an independent investigator in global oncology, focusing on strategies to enhance evidence-based breast cancer care delivery in resource- limited settings. This research aims to develop and evaluate strategies to improve healthcare provider adherence to clinical practice guidelines for breast cancer care in Nigeria. Candidate: Dr. Romanoff is a Breast Surgical Oncologist at the Icahn School of Medicine at Mount Sinai, where she is an Assistant Professor in the Department of Global Health and Health System Design, as well as a Global Cancer Disparities Researcher at Memorial Sloan Kettering Cancer Center (MSK). Aims for candidate development include education and training in: 1) fundamentals of health services research, 2) implementation science, and 3) global breast oncology. With the candidate development and research activities proposed, Dr. Romanoff will develop the tools necessary to become an independent investigator and leader in the field of global oncology, and meet her ultimate goal of improving breast cancer outcomes in resource-limited settings worldwide. Research: This research will use rigorous implementation science frameworks and methodologies to accelerate adherence to the NCCN Harmonized Guidelines for SSA in Nigeria, with the ultimate goal of reducing the evidence-to-practice gap and improving evidence-based breast cancer treatment. To achieve this, Dr. Romanoff will leverage the research infrastructure of the African Research Group for Oncology (ARGO), a research consortium including MSK and 26 Nigerian academic institutions. The overall hypothesis is that implementation of structured, multi-level strategies to enhance adherence to breast cancer care guidelines in Nigeria will be acceptable, appropriate, feasible and adoptable – and show promise to change clinical practice. Study aims are to: 1) identify multi-level barriers to guideline-concordant breast cancer care in Nigeria, 2) develop evidence-based implementation strategies to improve adherence to breast cancer clinical practice guidelines in Nigeria, and 3) rigorously evaluate selected implementation strategies in a pilot study in Nigeria. The rationale that underlies this proposal is that implementation of context-appropriate strategies to promote guideline adherence will improve breast cancer care delivery in Nigeria, and ultimately patient survival.

NIH Research Projects · FY 2024 · 2022-09

SUMMARY Therapies to treat Crohn's disease (CD) and ulcerative colitis (UC), the types of inflammatory bowel disease (IBD) characterized by severe chronic inflammation of the gastrointestinal tract, have greatly improved over the past three decades; yet, a large proportion of IBD patients fail to achieve sustained remission. Genetic biomarkers may offer an avenue toward the dissecting heterogeneity of disease etiology, and improving personalized therapeutic approaches. Our group has recently identified a coding gain-of-function mutation in the LRRK2 (leucine-rich repeat kinase 2) gene that conferred a ~70% increased CD risk and affected CD age of onset, disease location, LRRK2 kinase activity, and autophagy flux. Importantly, LRRK2 has also attracted considerable attention for its causal link to Parkinson's disease (PD). Further analyses identified dozens of additional variants within LRRK2 that were associated with the risk of both CD and PD, suggesting shared pathogenesis between these diseases. Given extensive efforts to target LRRK2 kinase activity as a means to treat PD, we explored known LRRK2 inhibitors developed for PD as potential therapies for IBD. We showed that selective LRRK2 inhibitors have ameliorated experimental colitis and reduced inflammatory cytokine TNF-α levels, a hallmark of IBD-associated inflammation, in dendritic cells of IBD patients. However, studies of LRRK2 kinase inhibition in preclinical models suggest that brain penetration and toxicity in peripheral tissues may be a critical safety liability for these inhibitors in IBD patients. To advance this therapeutic hypothesis, we have developed structure-based approach to design novel gut-restricted LRRK2 inhibitors, synthesized prototype inhibitors, crystallized analogs in complex with kinases mutants that mimic LRRK2 binding site and tested their LRRK2 potency and efficacy, which are maintained. Thus, we are proposing, starting from known LRRK2 inhibitor structures to 1) Develop proof-of-concept gut-restricted LRRK2 inhibitors as potential IBD therapeutics using structure-based drug design and medicinal chemistry; 2) Evaluate novel compounds for LRRK2 inhibition, in vitro absorption, distribution, metabolism, excretion and pharmacokinetics, off-target selectivity, and effects on cellular substrate phosphorylation and cytokine activity using biochemical and cell-based assays, and 3) Pre- clinically validate new inhibitors for in vivo absorption, distribution, metabolism, excretion and pharmacokinetics, plasma exposures, and effects on inflammatory biomarkers and severity of experimental colitis. Our hypothesis is that gut-restricted LRRK2 inhibitors could be a safe and effective therapeutic target of IBD therapy in the presence or absence of IBD-associated LRRK2 mutations. These studies will provide a basis for future clinical trials aimed at testing LRRK2 as an effective drug target for IBD. We believe our strong preliminary data and relevant therapeutic hypothesis align well with the research goals and objectives of PAR-19-294 and will help improve the current standard of clinical care for patients with this disease of interest to the National Institute of Diabetes and Digestive and Kidney Diseases.

NIH Research Projects · FY 2025 · 2022-09

Project Summary: Mouse genetic tools based on Cre recombinase and Cas9 nuclease have been applied for cell/tissue-specific gene knockout and cell lineage tracing. These tools enable control of gene expression and genome engineering to uncover detailed biology in a number of systems. However, the lack of precise spatiotemporal control hinders the broader applications of these tools. In this proposal, we will develop a new set of mouse genetic tools to address this issue. Then we propose to apply this system to investigate the establishment of the adult bone marrow hematopoietic stem cell (HSC) pool in vivo. Because HSCs self-renew and differentiate into all blood cell lineages throughout life, they are the basis for life-saving bone marrow transplantation in clinics. Therefore, understanding HSC biology has important translational implications. The establishment of the adult bone marrow HSC pool is essential for life-long hematopoiesis. However, how the adult bone marrow HSC pool is established has not been addressed. The lack of appropriate genetic tools to study HSCs at the clonal level in vivo has been a major hurdle. Our novel genetic tools will allow tracing HSCs at the clonal level in vivo. If successful, the results will significantly advance the field by directly addressing how the adult bone marrow HSC pool is established. Also, our proposed novel technologies will be widely applicable to various research fields beyond hematopoiesis to address fundamental questions.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY Diffuse midline gliomas (DMG), including diffuse intrinsic pontine gliomas (DIPG), are deleterious malignant pediatric tumors of the brainstem. DMG is the leading cause of death among pediatric brain tumors and the second most common malignant brain cancer afflicting children. DMG has a dismal prognosis of less than 1% survival within a year of diagnosis, even when using the most aggressive treatments. Approximately 400 children will be diagnosed with DIPG in the United States in 2022, all of whom will have a median survival of between 8 and 11 months. DMG disease outcomes have plateaued over the past decade due to the lack of effective treatments and limited diagnostic tools. Many failed clinical trials and therapeutic strategies in DMG can be attributed to two critical concerns: 1) the selectively penetrable blood brain barrier (BBB) restricts drug delivery to central nervous system, and 2) despite there being distinct genetic alterations between DMG and adult high- grade gliomas (aHGG), the agents considered for DMG clinical trials have been derived by extrapolation from aHGG data, without grounds for the therapeutic translation. Studies have revealed extracellular signal-regulated protein kinases (ERK), a downstream receptor tyrosine kinase of mitogen-activated protein kinase (MEK), is upregulated in DMG, raising questions about whether targeting the MAPK/ERK pathway can have anti-tumor effects in DMG. Targeting MEK in combination with aminolevulinic acid-photodynamic therapy (5-ALA-PDT) is of interest because inhibition of MEK has been found to significantly enhance protoporphyrin IX (PpIX) accumulation in vitro and in vivo in a tumor-specific manner. This proposal uses an innovative multimodal treatment approach that addresses the barriers to successful DMG clinical trials and exploits the molecular composition of DMG cells to reduce morbidity and mortality. By targeting MEK and employing 5-ALA-PDT, we anticipate MEK inhibition will synergize with 5-ALA-PDT efficacy by eliciting direct tumor cell killing, vascular shutdown and immune response, ultimately increasing overall patient survival. If successful, this treatment can be applied to other inoperable CNS tumors.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This proposal investigates statistical models for developing mobile health (mHealth) measures using patient generated health data (PGHD) with high complexity and temporality. The emergence of mHealth technologies and computational tools are rapidly expanding their use in research and clinical settings, and engaging patients in self-management. mHealth technology further allows integration of multifarious data streams to improve outcome measurement and prediction to aid clinical decision making. To maximize their actionability, however, there is a need to investigate novel approaches for design, development and evaluation of mHealth-based measures. We ground our investigation in chronic pelvic pain (CPP) as the disease model, a prevalent, complex disorder with high societal burden and quality of life (QoL) impact. There is substantial heterogeneity between patients and day-to-day variations in how CPP unfolds. Therefore, mHealth methods are particularly valuable for capturing the complex disease scenarios. There are no CPP-specific self-reported measures to assess disease status or treatment response. We propose to investigate models that can handle the inherent challenges of PGHD to derive ecologically valid and actionable self-tracking measures for patient outcomes in health settings. The Specific Aims are: Specific Aim 1. Investigate “critical windows of tracking” for mHealth-based disease outcome measurement. We will enroll 90 participants undergoing 12 weeks of physical therapy treatment for their CPP to use a mHealth app for tracking their symptoms, daily function, and medications. We will triangulate these data with clinician assessments and passive data on sleep and activity to build distributed lag models (DLMs) to identify predictors that can be used for outcome monitoring. Specific Specific Aim 2. Investigate a functional data analytic framework grounded in CPP to develop self- tracking pain and QoL measures. We will enroll 180 CPP patients to track their disease symptoms through a mHealth app and wear activity monitors for 3 months. Through a series of supervised and unsupervised models leveraging functional data analytic methods, we will identify variables to inform the design of the composite pain and QoL measures. Aim 2a. Design and develop a multidimensional self-tracking pain measure. We will build estimation models where the unit of observation is a set of curves (i.e., pain location, severity, type) over time, leveraging functional data analytic methods. Aim 2b. Design and develop a flexible self-tracking QoL measure. We will assess the relative predictive ability of individual items on CPP symptoms to derive a CPP-specific QoL measure that can be used at the day- vs week-level. Exploratory Aim 2: We will assess disease specificity of the models by comparing output from a non-CPP control group. Flexible, non- parametric data approaches allow maximizing the features of the available mHealth technology, which can aid in robust models to inform design of mHealth-based disease measures. Proposed work addresses the gap in mHealth evidence-base to improve the application and translation of efficacious mHealth assessments.

NIH Research Projects · FY 2025 · 2022-09

Project Summary T cell mediated graft rejection remains a critical barrier to long-term graft health and survival. Post-transplant complement-induced priming of T cells and the transfer of donor major histocompatibility complexes (MHC) to recipient dendritic cells (DCs) by graft-released exosomes (commonly referred to as “cross dressing”) are both mechanistically involved in the generation of anti-donor cellular immunity. Our preliminary data newly implicate mannose binding lectin (MBL) pathway-dependent complement activation as necessary mediator for complement opsonization of exosomes and exosome-mediated donor MHC delivery to recipient DCs. Together with our prior observation that recipient MBL pathway complement activation is required for generation of robust anti-donor T cell responses and costimulatory-blockade resistant graft rejection, these data lead to our central hypothesis that post transplant MBL pathway-initiated complement activation deposits complement opsonins on exosomes, which bind to recipient DCs via complement receptors, and that this process optimizes DC “cross dressing” to permit semi-direct pathway anti-donor T cell immunity and ultimately allograft rejection. We will test this hypothesis in two specific aims. In aim 1 we will study the mechanisms required for complement activation on exosomes, characterize the effect of complement opsonins on exosome binding to recipient DCs, and test the role of DC-expressed complement receptors in our model. In aim 2 we will test for links between complement-mediated DC “cross dressing” and post-transplant anti-donor T cell immunity and transplant outcomes. The findings will be significant because a) they will provide fundamental insights into the biology of exosome function, b) provide mechanistic links between exosomes, complement, and adaptive T cell immunity, and c) potentially identify novel treatment strategies and therapeutic targets to improve transplant outcomes. Our proposal is conceptually innovative and tests a novel paradigm linking exosome function and complement activation that may have broad implications beyond the field of transplantation.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Intervertebral disc degeneration is a major etiological factor of low back pain, which is the #1 cause of job disability worldwide that affects 80% of people at least once in their lifetime. Older individuals are prone to intervertebral disc degeneration by a loss of disc height and hydration. Pain can worsen intervertebral disc degeneration by limiting mobility and reducing the mechanical forces necessary for homeostasis. Therefore, there is an unmet need to safely protect older individuals from intervertebral disc degeneration and, here, we intend to repurpose existing bone therapeutics to reduce/prevent intervertebral disc loss and consequently back pain in the aging population. Intervertebral disc degeneration remains a growing problem because (1) the US geriatric population is projected to outnumber children for the first time in history, (2) the early treatment of intervertebral disc degeneration remains palliative and (3) the ballooning socioeconomic burden. We are uniquely positioned to bridge the knowledge gap in the molecular mechanisms of bone therapeutics to stem intervertebral disc degeneration. Aging and mechanical injury of the intervertebral disc by compression both exacerbate breakdown of the extracellular matrix, but compression stimulates inflammation more so than aging. By contrast, we recently found that deletion of the inhibitor of Wnt/β-Catenin signaling sost, a bone formation anabolic mechanism and the genetic analog approach to anti-sclerostin antibody injection, increases the structural properties and hydration of the intervertebral disc. Another bone therapeutic that shows benefits to the intervertebral disc is anti-resorptive raloxifene, a non-uterine-targeting estrogen agonist, that also increases binding of water to collagen. Use of raloxifene in postmenopausal women is associated with relieving back pain and greater intervertebral disc height than non-treated women. Our studies show that raloxifene increases intervertebral disc height in male and female mice, reduces sex- and age-related intervertebral disc degeneration in female mice, improves metrics of pain-related behavior in old mice and stimulates Wnt and estrogen signaling. Here, we hypothesize that currently available therapeutics will stimulate extracellular matrix anabolism and prevent compression-induced intervertebral disc degeneration in young and aged mice. To test this hypothesis, we propose two aims using aged mice and catabolic compression as models of intervertebral disc degeneration. In aim 1, we will determine whether therapeutics known to activate Wnt signaling will prevent IVD degeneration by injurious mechanical compression in young and aged mice. In aim 2, we will determine whether estrogen signaling prevents IVD degeneration via Wnt signaling and whether prevention can be safely improved by combining the signaling pathways in aged mice. This proposal will repurpose current FDA-approved bone prophylactics that target pathways consistently impacted by intervertebral disc degeneration in variable conditions and may be specifically beneficial to intervertebral disc degeneration in the elderly.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY: West Nile virus (WNV) is a highly virulent human pathogen of the central nervous system (CNS) and the most common cause of epidemic encephalitis in the United States. There are no vaccines or specific antiviral treatments available for WNV-infected individuals, and development of such an arsenal requires a more complete understanding of its pathogenesis. WNV is primarily transmitted through the bit of an infected mosquito. Once deposited into the skin, the virus replicates in a local draining lymph node prior to viremia, which allows the virus to spread to various organs, including the CNS, where neurons are the major targets of infection. Currently, it is unclear which cells are infected outside of the CNS, but numerous in vitro studies suggest that myeloid cells, such as monocytes/macrophages, are likely the primary targets. To begin understanding this in a more physiologically relevant model, we utilized an ex vivo human lymphoid tissue model, where WNV replicates robustly. We immunophenotyped the WNV-infected cells and found that a significant proportion were CD4+ T cells. To further validate these findings, we generated WNV mutant strains that contain cell-specific microRNA (miR) targets (miR-Ts) to restrict WNV replication in a cell-specific manner. We engineered lymphoid- or myeloid-specific miR-Ts into the genome of WNV and found that WNV strains unable to replicate in myeloid cells were able to replicate nearly identically to wild-type WNV strains, while WNV strains incapable of replicating in lymphoid cells were unable to replicate in this same model. Therefore, we hypothesize that lymphoid cells are essential initial cellular targets for WNV replication in the periphery. In this application, we seek to identify the cells that are essential for WNV replication in the human lymphoid tissue ex vivo (Aim 1) as well as in mice (Aim 2).