Icahn School Of Medicine At Mount Sinai

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Social interaction strongly influences both momentary mood and general mental health. Indeed, social isolation during the recent pandemic caused record-high levels of mood symptoms in many individuals around the world. This relationship between social interaction and mood has been documented not only in mood disorders, but also in the descriptions of many other psychiatric illnesses. So, better elucidating the influence of social interaction on mood may have powerful implications for treating those burdened by socio-affective deficits. Recent work in computational psychiatry has defined mood as a progressive impact of reward prediction errors (RPEs), which reflect the differences between predicted and perceived rewards. These efforts have been expanded to account for learning specifically within the social domain. In society, we must often learn social norms, or agreed-upon rules of behavior among social groups. These norms change over time, as driven by norm prediction errors (nPEs), which act as learning signals that help us to predict norms more accurately in the future. nPEs, like RPEs, influence short-term mood. Yet, it is not well known how nPEs relate to long-term mood symptoms. We are also limited in our understanding of how nPEs are encoded in the brain. Prior work has predominately relied on neuroimaging techniques, which can identify high-level cortical circuits involved in norm prediction but cannot as reliably assay the potential involvement of subcortical micro-structures. Alternatively, human single-unit recording may be used to directly examine the roles of micro-structures in norm prediction at the resolution of single neurons. Single-unit studies have identified neurons in the substantia nigra (SN), a subcortical nucleus that is hard to reach with fMRI, that are involved in RPE encoding. Given similar computational relationships with mood and associated activity on neuroimaging, RPEs and nPEs may also share common encoding by SN neurons. To date, this hypothesis has never been tested. Considering these critical knowledge gaps, the objective of this proposal is to investigate the neurocomputational mechanisms underlying the influence of social interaction on mood by leveraging computational modeling (Aim 1) and human single-unit recording (Aim 2). The central hypothesis is that nPEs influence both short- and long-term changes in mood; we further predict that nPEs are neurally encoded by single-units in the human SN, which is supported by pilot data collected from twelve patients so far. These findings will expand our neurophysiological understanding of the relationship between mood and social learning in health and disease in hopes of informing future psychotherapeutic interventions for disorders that adversely affect social and affective functioning. This research will take place at the Centers for Computational Psychiatry and Advanced Circuit Therapeutics at the Icahn School of Medicine at Mount Sinai. Completion of this proposal will afford the applicant rigorous training in computational modeling and human single-unit recording, thereby accelerating her progress towards becoming a future physician-scientist and leader of a research program in invasive computational psychiatry.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Violations of our expectations result in prediction-error signals in the brain. When these prediction errors are repeatedly experienced, they can influence our current affective state, mood, and ultimately behavior. Despite this understanding, the brain circuits and patterns of activity within the circuits that translate these prediction error signals into affective state fluctuations is unknown. Uncovering this is fundamental to gaining insight into the neural basis of moods in healthy people, what becomes dysfunctional in pathological states like depression, and for the development of new treatments that focally target these circuits. The experiment in this proposal will combine high-density, multi-region, neurophysiology with state-of-the-art computational models to determine neural and physiological biomarker of mood-like states in macaques. During the K99 phase of this proposal, I will train with Drs. Rudebeck (primary mentor), Mayberg (co-mentor) at the Icahn School of Medicine at Mount Sinai and Dr Rutledge (co-mentor) at Yale to test the hypothesis that a recent history of negative or positive prediction errors will cause changes in affect through different subgenual cingulate cortex circuits that are specific to the type of prediction violation encountered. This will be achieved by combining computational models of affect with the simultaneous recording of activity across multiple limbic and prefrontal brain circuits while awake non- human primates perform multiple different reward-based tasks that manipulate the recent history of prediction errors. Next, I will focally target activity in these brain circuits using non-invasive transcranial focused ultrasound aimed at subgenual cingulate cortex while concurrently recording activity from its interconnected brain areas. The NIH K99/R00 Pathway to Independence Award will give me the unique opportunity to receive a multi- disciplinary and technical training under the K99 portion. This will include training in the acquisition and analysis of single-neuron and population activity, computational modeling, and how insights from basic research can influence the clinic. In addition, during this phase I will receive career-shaping training in how to lead my own laboratory (i.e. managing people, writing a grant and networking), translate research from the laboratory to clinical settings, apply to and obtaining a tenure rack position at a top-tier academic research institution. In the R00 phase of the proposal I will combine neurophysiology with transcranial focused ultrasound to test the hypothesis that causal manipulation of the history of reward prediction error in subgenual cingulate cortex stabilize affective state fluctuations. The work proposed will provide strong preliminary data on the neural and computational mechanisms of affect and a non-invasive ultrasound protocol to modulate neural activity within these circuits data. This will be foundational for my lab’s first grant applications and to establish myself as a successful principal investigator causally investigating the neural basis of affective states in healthy brains as well as in disease.

NIH Research Projects · FY 2025 · 2024-07

Summary The mechanosensory Merkel cells (MCs) are crucial for encoding the sense of light touch. Recent studies have shown that MCs also significantly regulate alloknesis, the itch sensation caused by light mechanical stimulation. Notably, alloknesis is commonly observed in the elderly, with research indicating a decrease in MC numbers in aged human and mouse skin. Among many skin abnormalities occurring with aging, dry skin condition appears to be involved in alloknesis. In experiments using a mouse model of experimental dry skin induced by acetone- ether-water (AEW) treatment, a decline in MC numbers and an increase in alloknesis were observed. Interestingly, upon discontinuing the AEW treatment, MC numbers returned to normal, and the mechanical itch was gradually alleviated. This suggests that uncovering the cellular and molecular mechanisms controlling MC decrease or MC regeneration could pave the way to designing therapies to prevent alloknesis in aging and dry skin. In Aim 1, we will focus on understanding the mechanisms behind the regeneration of MCs. Pilot studies showed that MC recovery is not due to MC proliferation; however, the exact source of MC regeneration remains unknown. We demonstrated that the Tenascin C (TNC)-expressing epidermal keratinocytes are MC progenitors. Here, we will use lineage tracing and single-cell RNA sequencing to investigate if TNC-positive cells are responsible for MC regeneration upon AEW treatment and to identify cellular and molecular processes controlling MC regeneration. We will then perform genetic studies to examine the functional significance of identified processes on MC regeneration. Finally, we will analyze molecular changes in MC progenitors and the MC differentiation program in young vs old mice and uncover the functional significance of the identified changes on the age-related decline of MCs. In Aim 2, we will investigate the interaction between MRGPRA3/TRPV1+ itch- initiating C fibers and MCs, as our studies showed that the ablation of MRGPRA3/TRPV1+ neurons prevents MC reduction upon AEW treatment. Here, we will perform chemogenetic manipulations of the activity of MRGPRA3+/TRPV1+ sensory neurons to reveal the causal relationship between sensory neuron activities and MC numbers. We will further identify molecules secreted by MRGPRA3/TRPV1+ neurons post-AEW treatment and receptors of these molecules in MCs or MC progenitors, shedding light on the cellular and molecular dynamics controlling MC numbers. We will test the functional significance of the identified ligand-receptor pairs for controlling MC numbers and inducing alloknesis by performing functional genetic studies in the settings of aging and dry skin. Overall, these studies aim to offer new insights into preventing age-related MC decline and associated alloknesis, potentially leading to novel therapeutic interventions.

NIH Research Projects · FY 2020 · 2024-07

Summary Mammals have formed an evolutionary alliance with the commensal microbes that inhabit our epithelial boundaries. This lifelong relationship is forged at birth when microbes first colonize our surfaces. Alterations in neonatal microbial communities are associated with several prevalent epithelial skin dysfunction interactions inflammatory diseases, including atopic dermatitis. Atopic ermatitis is a devastating disease affecting about 20% of children in the western world and is rooted in epidermal barrier and commensal dysbiosis. Yet, surprisingly little s known about the functional between colonizing commensals and epithelial populations in neonatal skin. d i Epidermal stem and progenitor cells (ESPCs) maintain the epidermis throughout our lifetime by taking cues from the microenvironment or “niche”. We recently uncovered a remarkable capacity for ESPCs to sense, respond to, and remember inflammatory stimuli. Whether and how commensal signals similarly influence ESPCs and shape epidermal tissue fitness is an open question. Here we address the tantalizing possibility that commensal microbes are a heretofore unappreciated ESPC niche component. Thus, we aim to illuminate their roles in directing ESPC differentiation, innate immune activation and, consequently, epithelial barrier function and fitness in neonates, when microbes first colonize the skin. We also seek to functionally examine how the early-in-life epithelial–microbe dialogue impacts atopic disease susceptibility. Our proposed use of genetically tractable commensal strains and epidermal-specific deletion of defined microbial sensors with a state-of-the art in vivo tissue-specific gene modulation system to systematically manipulate both host and microbe allows for a comprehensive and mechanistic understanding of the microbe–ESPC dialogue and its impact on epidermal health and disease. The findings the generated from these studies will lay the groundwork for developing microbiota-based therapies to boost the epidermal barrier and mitigate atopic disease.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY In the first 6 months after COVID-19 vaccine approval, 3.4 billion doses were administered globally, potentially saving 7.2 million lives. Lipid nanoparticle (LNP)-mRNA based COVID-19 vaccines have been able to induce both a strong T cell- and antibody-mediated responses against SARS-CoV-2 by triggering myeloid antigen presenting cells (APCs) such as dendritic cells (DCs), demonstrating the efficacy of LNP-mRNA technology to modulate the immune system. Partly due to the success of COVID-19 vaccines, there has been a renewed attention in engineering novel LNP formulations to better modulate the myeloid APC compartment by 1) either fine-tuning the adjuvanticity of the LNP lipid formulation or 2) improving in vivo targeting efficiency of LNPs towards myeloid APCs. Although the advancements in LNP lipid chemistry enrich our arsenal of LNP formulations suited to modulate the immune system, there hasn't been a corresponding detailed understanding of the molecular and cellular changes induced upon the uptake of LNPs in specific cell types and tissues. In fact, attempts to elucidate the innate mechanisms of LNP immunogenicity have often led to conflicting conclusions and have in turn suggested that the mechanisms of LNP immunogenicity are rather LNP formulation-dependent. The lack of consensus on the innate mechanisms of LNP immunogenicity further highlight that the immunogenicity of each LNP is a complex function of a) its lipid formulation and the nature of its payload, b) the pattern recognition receptor (PRR) profile unique to each immune subset, c) LNP targeting efficiency in each cell type and d) cell, tissue and disease context of LNP treatment. Without the parallel efforts to gain mechanistic understanding of immunogenicity of each LNP formulation, it remains difficult to rationally select LNP formulations that are tailored to a particular disease context, including cancer. In using LNPs as a cancer immunotherapeutic, my central hypothesis states that LNP lipid formulations that drive up their immunogenicity combined with an engineering strategy to increase specific targeting of DCs in vivo can synergize with immunostimulatory payloads (i.e. IL-12) to reprogram DC cell states and enhance anti-tumor immunity. In Aim 1, I will characterize the changes in DC phenotype, function and transcriptional profile upon uptake of DC-targeted LNPs in both steady state and tumoral contexts using DC-T cell co-cultures, bulk and scRNA-seq of sorted LNP transfected DCs. In Aim 2, I will engineer Clec9a targeted bi-specific antibodies to improve LNP transfection of DCs in vivo. The outcome of this project is both a mechanistic understanding of the interplay between LNPs and DCs that occurs upon LNP uptake and a novel bi-specific antibody-based strategy to increase DC targeting in vivo.

NIH Research Projects · FY 2025 · 2024-07

Project Summary The approval of anti-PD1/PD-L1 and anti-CTLA immune checkpoint blockade (ICB) therapies for those with non- small cell lung cancer (NSCLC) has demonstrated that immunotherapies can generate robust responses for a subset of patients with advanced disease. However, while nearly 45% of patients demonstrate a major pathological response to ICB, the median progression-free survival for NSCLC patients remains dismal at around 7 months. It is now well documented that persistent interferon gamma (IFNy) cytokine pathway signaling plays a paradoxical role within the tumor microenvironment and has been associated with both resistance and response to ICB leading us to believe that there is greater nuance to dissect within this pathway. Accordingly, this cytokine pathway is quite complex as IFNy stimulates the expression of ~200 hallmark interferon-stimulated genes (ISGs); thus, it is still unclear how the majority of ISGs influence tumor immunity as it has been difficult to study this pathway in a relevant biologic context. Using a novel functional genomics platform developed in our lab (Perturb-map) my sponsor lab was able to knock-out (KO) 35 cytokine genes in parallel in the KP mouse model of NSCLC and investigate spatial and temporal phenotypes at a single-cell resolution. It was found that KO of Socs1, a negative regulator of the IFNy pathway, caused an overt growth advantage and a paradoxical increase in both CD8+ and CD4+ T-cell infiltration into tumors. Further, these tumors were found to be more sensitive to PD-L1 blockade than wildtype tumors. We hypothesize that Socs1 KO causes chronic upregulation of tumor cell IFNy signaling leading to cytokine-mediated infiltration of the TME which is followed by an increase in immune-inhibitory signal expression on tumor cells such as PD-L1. In Aim 1, we will investigate the influence of Socs1 KO on tumor immune-composition of the TME and tumor phenotype using Perturb-map and transcriptomics. We will then corroborate these data with CITE-seq of human lung cancer specimens to further explore the role of SOCS1 in human malignancy. In Aim 2, we will use Perturb-map to KO all 162 ISGs and determine how each impacts tumor immunity in the context of immune checkpoint blockade therapy and TME biology at depth. These experiments will include assessment of immune & stromal recruitment, tumor organization & differentiation, and unbiased analysis by spatial transcriptomics to identify mechanistic programs controlled by each ISG.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Psychiatry faces a significant challenge in the absence of objective measures to assess behavior. Clinicians form clinical opinions based largely on their impressions from interviewing and what they read in the electronic health record. As a result, we are currently unable to provide reliable prognoses on an individual basis. One untapped source of behavioral information for clinical decision-making is the clinical interview itself, which forms the foundation of the electronic health record (EHR). Every clinical visit provides a wealth of behavioral information comprising spoken language, eye contact, and facial expressions from both the patient and the clinician. Another source of behavioral data, which is ecologically valid, comes from smartphones, which provide physical activity metrics (e.g., step count, distance traveled), geolocation, social interactions (e.g., SMS messages and phone calls made and received), sleep patterns and audio data from diaries. By analyzing these rich behavioral datasets from routine clinical visits and smartphones, we can develop clinical signatures for particularly clinically relevant outcomes in young help-seeking people, namely treatment disengagement, ER visits and hospitalizations. These individualized clinical signatures are important for the real-life situation that confronts both clinician and patient at the first visit to a mental health clinic. This proposal includes all new patients (N = 2100), ages 15 to 30, who seek treatment for the first time at one of six outpatient mental health clinics in the Mount Sinai Health system. Aim 1 is to create a baseline clinical signature for outcomes using deep neural network modeling of legacy EHR data and baseline behavior, which includes audiovisual recordings of intake interviews, ratings of working alliance, and brief surveys and tests of cognition. Aim 2 is to use Contextual Bandit to create a longitudinal clinical signature for outcomes based on subsequent behavioral data from clinical interviews (and their accompanying notes), and smartphone passive data and audio diary data. Contextual Bandit is a model that keeps updating probabilities and odds over time as it is given new data. Aim 3 is to create clinical signatures based on EHR data alone, such that the added value of behavioral data for Aims 1 and 2 can be quantified. Study assessments are standard, low-cost, and easy to administer, with good variance, validity, reliability, and generalizability. Across all aims, fusion will be used for behavioral feature extraction and natural language processing (NLP) for analysis of both written language (clinical text) and spoken language (clinical visits and audio diaries). Data science methods have been optimized for partnership with the DCC. Community engagement and ethical issues re privacy, informed consent and fairness have been prioritized.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Psychiatry faces a significant challenge in the absence of objective measures to assess behavior. Clinicians form clinical opinions based largely on their impressions from interviewing and what they read in the electronic health record. As a result, we are currently unable to provide reliable prognoses on an individual basis. One untapped source of behavioral information for clinical decision-making is the clinical interview itself, which forms the foundation of the electronic health record (EHR). Every clinical visit provides a wealth of behavioral information comprising spoken language, eye contact, and facial expressions from both the patient and the clinician. Another source of behavioral data, which is ecologically valid, comes from smartphones, which provide physical activity metrics (e.g., step count, distance traveled), geolocation, social interactions (e.g., SMS messages and phone calls made and received), sleep patterns and audio data from diaries. By analyzing these rich behavioral datasets from routine clinical visits and smartphones, we can develop clinical signatures for particularly clinically relevant outcomes in young help-seeking people, namely treatment disengagement, ER visits and hospitalizations. These individualized clinical signatures are important for the real-life situation that confronts both clinician and patient at the first visit to a mental health clinic. This proposal includes all new patients (N = 2100), ages 15 to 30, who seek treatment for the first time at one of six outpatient mental health clinics in the Mount Sinai Health system. Aim 1 is to create a baseline clinical signature for outcomes using deep neural network modeling of legacy EHR data and baseline behavior, which includes audiovisual recordings of intake interviews, ratings of working alliance, and brief surveys and tests of cognition. Aim 2 is to use Contextual Bandit to create a longitudinal clinical signature for outcomes based on subsequent behavioral data from clinical interviews (and their accompanying notes), and smartphone passive data and audio diary data. Contextual Bandit is a model that keeps updating probabilities and odds over time as it is given new data. Aim 3 is to create clinical signatures based on EHR data alone, such that the added value of behavioral data for Aims 1 and 2 can be quantified. Study assessments are standard, low-cost, and easy to administer, with good variance, validity, reliability, and generalizability. Across all aims, fusion will be used for behavioral feature extraction and natural language processing (NLP) for analysis of both written language (clinical text) and spoken language (clinical visits and audio diaries). Data science methods have been optimized for partnership with the DCC. Community engagement and ethical issues re privacy, informed consent and fairness have been prioritized.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Primary sclerosing cholangitis (PSC) is a chronic inflammatory condition of the biliary epithelium characterized by periductal fibrosis, a heightened risk for cholangiocarcinoma, and frequent comorbidity with inflammatory bowel disease (IBD). The periductal fibrosis observed in PSC narrows the lumen of affected bile ducts and causes extrahepatic cholestasis, which mediates further liver injury. PSC patients typically develop end-stage liver disease within 10-20 years of diagnosis, making PSC a clinically significant cause of liver transplant in the United States. There are currently no disease modifying drugs for PSC, underscoring the unmet need for novel research that characterizes the pathologic mechanisms that drive periductal fibrosis in PSC patients. Aim 1 seeks to dissect the cellular heterogeneity of the periductal niche and characterize cell subpopulations mediating periductal fibrosis in PSC patients using single-cell multiomics. In this aim, the 10X Genomics Next GEM single-cell RNA-sequencing platform will be used to perform integrative analysis of gene expression and chromatin availability in single cells. The gene expression signatures of PSC patient tissue and PBMCs will be compared to patients with metabolic dysfunction-associated steatohepatitis (MASH) and non-cirrhotic controls. Aim 2 seeks to map the spatial organization of cell subpopulations within PSC periductal lesions and reveal cellular interactions within intact tissue. In this aim, the 10X Genomics Xenium in situ Gene Expression Analyzer will be employed to measure the expression of 400 genes in 10µm thick, 12mm x 22mm tissue sections produced from explanted liver tissue. This work will shed new light on the cellular landscape that drives periductal fibrosis, biliary-type cirrhosis, which is often present in patients with advanced PSC, and MASH cirrhosis, which is included as a control. Aim 3 seeks to characterize the effects of a leaky biliary epithelium on macrophage function and collagen production in the periductal niche using an in vitro model. Bile-stimulated macrophages will be characterized at the RNA and protein level and differences in the activating potential of PSC patient bile versus control bile will be assessed. Macrophage-fibroblast co-cultures will also be treated with bile and collagen production will be measured using the Sircol assay. This work utilizes patient-derived bile samples and an in vitro myeloid- stromal co-culture model to resolve luminal, myeloid, and stromal contributions to periductal fibrosis. Mount Sinai Hospital cares for one of the largest and most diverse PSC patient populations in the United States, and the Icahn School of Medicine home to pioneers in the field of hepatic fibrosis research. The training plan for this fellowship takes full advantage of the academic environment at Mount Sinai, and the research strategy combines a single-cell approach with the development of a novel cell culture model to assess cellular drivers of periductal fibrosis. This work will characterize the cellular landscape of periductal fibrosis at a single- cell resolution and promote the development of novel diagnostics and therapeutics for PSC patient care.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Down syndrome (DS) affects 1 in 700 newborns in the United States. It is the most common genetic cause of intellectual disability. DS individuals also frequently present with cardiac anomalies and are at increased risk for Alzheimer's disease, hematological neoplasia, autoimmune conditions (e.g., hypothyroidism, celiac disease, type I diabetes, arthritis), and inflammatory skin diseases such as atopic dermatitis (AD), alopecia areata (AA), and vitiligo, but also psoriasis and hidradenitis suppurativa. Studies have shown that the inflammatory and autoimmune comorbidities associated with DS may be due to overt inflammation caused by increases in many cytokines which signal through the JAK/STAT pathway. This inflammation may also be due to increased presence of type I, II and III interferon receptors, all encoded on chromosome 21, which also use JAK/STAT pathway to signal. Treatment of inflammatory skin diseases in individuals with DS involves broad immunosuppressive medications, which are often associated with multiple side effects and limited efficacy. Given the term hyperactivation of JAK/STAT signaling in DS, JAK inhibition presents an appealing alternative for better long- disease control.Several case reports, case series, and studies across inflammatory joint and skin conditions showed the efficacy, safety and tolerability of the pan-JAK inhibitor tofacitinib in individuals with DS. Importantly, none of these studies reported new safety concerns. Further, more selective JAK inhibitors, such as the JAK1/JAK2 inhibitor ruxolitinib, demonstrated potential in synergistically ameliorating hematologic conditions associated with DS. This is consistent with our own studies, in which ex vivo tofacitinib successfully attenuated hyperinflammation in individuals with DS. However, while tofacitinib is approved in rheumatoid arthritis, it has not been FDA-approved for any skin indications (risk was determined by the FDA to outweigh benefit in psoriasis), and nor has oral ruxolitinib. Thus, individuals with DS deserve timely and appropriate evaluation of the long-term safety and efficacy of newer and more specific JAK inhibitors that are currently FDA- approved for dermatological indications, such as AD and AA. Since a particularly higher inflammatory tone has been shown in DS, the dosing of these newer drugs and their long-term safety and efficacy should also be evaluated in patients with DS. Such drugs include abrocitinib, a JAK1 inhibitor that was recently approved for moderate-to-severe AD in both typical adults and typical adolescents aged at least 12 years, and ritlecitinib, a JAK3/TEC inhibitor that has completed phase 3 studies in AA and has just obtained FDA approval in AA. Herein we propose a clinical trial to evaluate the long-term safety and efficacy of 60 weeks of treatment of the low and high dose of abrocitinib in individuals with DS presenting with AD and AA, and of ritlecitinib in those individuals with AA that did not adequately respond to abrocitinib within the first 24 weeks of treatment.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY / ABSTRACT There is a fundamental gap in understanding how neurons integrate hypothetical information when making decisions. This includes not only predicting future outcomes before making choices but also evaluating consequences of unselected alternatives. These computations serve as the basis of counterfactual thinking and regret processing. This phenomenon is central to a wide range of cognitive capabilities but whose dysfunction, despite contributing to numerous psychiatric disorders, is poorly understood. My long-term goal is to uncover the mechanisms governing counterfactual thinking, how the brain binds hypothetical value to unselected actions, and how this translates into changes in motivated behavior. This proposal will determine how single neurons in the nucleus accumbens (NAc) – a critical node of value integration and action selection – support counterfactual thinking constrained by its known inputs. We will leverage innovative approaches in rodent neuroeconomics that we developed to capture complex, evolutionarily conserved decision-making processes across species. We will combine this with cutting-edge brain-wide imaging tools and novel circuit-dissection technologies we developed to measure circuit physiology at an unprecedented level. Our central hypothesis is that populations of functionally distinct neurons in the NAc defined by upstream inputs are differentially involved in assigning credit to missed reward-related opportunities due to different unselected actions. This hypothesis is based on our preliminary data implicating multiple circuits that converge in the NAc and may play distinct roles in action-specific forms of counterfactual thinking. This hypothesis will be tested by pursuing two specific aims: 1) Characterize hypothetical value encoding of NAc neurons defined by their upstream inputs; and 2) Establish a link between afferent activity in the NAc and action-specific forms of counterfactual thinking. First, single-cell NAc activity categorized by the structures projecting to them will be recorded during decision-making behavior in mice. We will characterize firing properties following economic situations known to invoke representations of missed opportunities and will manipulate their excitability using chemogenetics to alter the impact of decision history on future behavior. Second, we will record activity of axonal afferent fibers from major excitatory inputs into the NAc simultaneously with single NAc cells and use optogenetics to manipulate input terminals during distinct action-selection processes. This approach is innovative because it captures input-output circuit physiology in ways never before measured in freely behaving animals using our newly engineered open two-color Miniscope. This is significant because it answers a fundamental biological question: how do single neurons integrate hypothetical value and assign credit to unselected actions? Furthermore, this work will test competing theories whether counterfactual thinking derives from a generalized computation for mistake appraisal or instead comprises dissociable, action- specific processes. Ultimately, this work has the potential to refine our diagnostic nosology of and develop circuit- based treatments for how emotion-cognition interactions like regret processing go awry in psychiatric disorders.

NIH Research Projects · FY 2025 · 2024-07

Asthma is one of the most common physical health conditions in World Trade Center (WTC) responders, affecting approximately one third of individuals. Moreover, most WTC responders with asthma continue to report substantial symptoms, experience poor disease control, and have high acute resource utilization and healthcare-related costs. Thus, asthma remains a major cause of morbidity and compromised quality of life in this population. Management of asthma in WTC-exposed individuals can be challenging. High prevalence of physical comorbidities (e.g., chronic rhinitis, obstructive sleep apnea) and mental health problems (e.g., post- traumatic stress disorder, depression) that contribute to increased asthma morbidity are common among WTC responders. Active self-management is critical for achieving adequate long-term asthma control. Unfortunately, low adherence to self-management behaviors (SMB) is a common problem and may involve unique pathways (e.g., WTC-specific disease and medication beliefs) in WTC responders. Cognitive decline due to aging and WTC exposures can further complicate asthma self-management. The complex constellation of medical, cognitive, and mental health conditions accompanying asthma in WTC responders necessitates comprehensive self-management support (SMS) interventions to improve their outcomes and quality of life. Unfortunately, there are no available interventions to improve the outcomes of this population. We previously developed and successfully tested the Supporting Self-Management Behaviors for Adults (SaMBA) program. The SaMBA model of SMS is an evidence-based approach grounded in a theory of health behavior that is unique among chronic illness SMS programs because its strategy involves comprehensively screening for barriers to effective self-management and disease control and couples it with tailored, theory-based, barrier- specific interventions. In this study, we propose to adapt the SaMBA model to the needs of WTC responders with asthma. Our Specific Aims are to: 1) Enhance and adapt an asthma SMS model for WTC responders with asthma (SaMBA-WTC), with input from WTC responders and other stakeholders to validate content and protocols and 2) Conduct a pilot RCT of SaMBA-WTC to determine feasibility and preliminary impact, in preparation for a fully powered trial. We will conduct a pilot RCT (n=58) of the SaMBA-WTC model delivered to WTC responders for 3 months vs. a time-matched asthma education attention control. Data will be collected at the end of the 3-month intervention (3-month visit) at 3 months later (6-month visit to assess sustainability). Clinical outcome measures will include asthma control, quality of life, resource use and measures of asthma self-management. The intervention is significant for its attention to the full range of factors that contribute to suboptimal self-management and poor outcomes among WTC responders with uncontrolled asthma. It is innovative for its comprehensive and theory-based screening approach to identifying and addressing coping strategies and barriers to effective SMB in asthma in this specific population.

NIH Research Projects · FY 2024 · 2024-07

PROJECT SUMMARY Effective mentorship is a critical component for academic and career development, particularly for trainees underrepresented (UR) in the biomedical workforce. The Mount Sinai Post-Baccalaureate Research Education Program (PREP) has a long history of successfully training UR scholars for biomedical graduate programs (e.g., PhD or Md-PhD). This important work requires dedicated time and effort to adequately support the unique needs of UR trainees, who may need more support and attention. Substantial data reveals that racial and other biases persist in biomedical education. For example, a 2022 Pew Research study found that nearly half of all Black workers in STEM (48%) experienced mistreatment in their STEM education, with over one-third reporting that someone treated them as if they could not understand scientific concepts (34%) or made them feel as though they did not belong in STEM (34%). There is growing recognition that higher rates of attrition among racial/ethnic graduate students will hamper efforts to diversify our biomedical workforce – efforts that are critical to our ability to conduct high-level science and deliver optimal health care. Cultivating a culture of inclusivity and psychological safety within the research training environment are important components to developing a diverse biomedical workforce. Thus, the goal of the proposed supplement is to enhance the Mount Sinai PREP in two ways: 1) well- being and psychosocial support for trainees and 2) additional anti-racism and -bias training for faculty mentors. The proposed activities will provide unmet psychosocial support for our PREP trainees by offering opportunities to discuss their mental and emotional well-being as URM trainees, and mentor training specific to mitigating racism and racial stereotypes for program faculty. We will accomplish these goals by partnering with faculty from the Mount Sinai Center for Scientific Diversity (CSD) who will lead monthly group discussions with PREP trainees on relevant well-being topics including scientific identity, sense of belonging, imposter phenomenon and resilience. Additionally, PREP faculty mentors will participate in a 5-week mentor training workshop (1.5-hour sessions) lead by CSD faculty. The workshop curriculum will teach mentors practical strategies for mitigating racism and racial stereotypes in their research labs as well as within their mentor-mentee relationships. Outcomes will be evaluated at the individual level (i.e., PREP trainees and faculty mentors) and program level (e.g., number of participants) for each component (e.g., mentor training and well-being discussion sessions). Our long-term goal is to impact the research experiences of our PREP trainees and the overall effectiveness of our PREP program. If proven successful, we will obtain institutional support to sustain the proposed activities in both Mount PREP and across all research training programs within the institution.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY The relatively finite number of ~20,000 protein coding genes in the mammalian genome limits the overall diversity that transcriptional regulation can achieve in a given biological context. This diversity can be enhanced by many orders of magnitude by post-transcriptional regulation such as alternative splicing and modification and degradation of mRNA, which in turn leads to a greater protein diversity and thus a greater functional complexity of the cell. This seems particularly relevant in the context of complex processes such as cell specification and organogenesis. The importance of RNA regulation is further supported by the large number RNA-binding proteins encoded in the human genome (~1,500) and their ever more prominent emergence in the context of disease. Despite the clear evidence of the relevance of RNA regulation, much less remains known compared to the exceptionally well-researched gene regulatory mechanisms. This lack of progress is in part due to technical challenges associated with studying and manipulating RNA, particularly during early development. However, recent advancements in this area now afford new opportunities to broaden our knowledge in these critical regulatory mechanisms and start to fill this critical knowledge gap. Here we will interrogate RNA-binding proteins important for cardiac development, specifically the the X-linked protein DDX3X. Mutations in DDX3X lead to DDX3X Syndrome, which is characterized by intellectual disability, autism spectrum disorder, congenital brain malformations and motor problems. DDX3X has not been interrogated during cardiogenesis to date, but recent studies have uncovered that individuals with DDX3X Syndrome frequently present with congenital heart disease. Based on our preliminary findings we hypothesize that DDX3X acts in a spatio-temporal manner, by targeting distinct regulatory networks during the development of the different cell types of the heart. We further hypothesize that disease-causing mutations in DDX3X affect cellular dysfunction by specific mechanisms of DDX3X target regulation. Aim 1 will uncover the direct, and functionally relevant targets of DDX3X. Aim 2 will determine the cellular phenotype of DDX3X-deficient male and female embryos. Aim 3 will elucidate genotype-phenotype correlations in DDX3X Syndrome patient-derived hPSC-CMs. Successful completion of the aims are expected to identify and characterize the cellular requirements and in depth molecular mechanisms of DDX3X, a new RNA-binding protein that we found to be essential for heart development. The overall and long-term goal is to contribute to a better understanding of RNA regulatory mechanisms, specifically during cardiac development and in the context of the formation of congenital heart disease.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Our long-term goal has been to characterize the heterogeneous group of chronic lower airway diseases (LAD) observed in World Trade Center (WTC) workers and volunteers, uncover their risk factors and comorbidities, identify subgroups with adverse and favorable lung function trajectories and outcomes, deploy novel imaging approaches to the investigation of the underlying lung injury, and improve surveillance and prevention strategies. Our previous research suggests the differential adverse effects and imaging characteristics of the longitudinal lung function trajectories of the WTC workers, and markers suggesting early chronic disabling lung disease. We thus propose to test the use of added respiratory surveillance tools and explore functional markers of disease progression, explore alternate methods to investigate longitudinal functional trajectories, and novel spirometry calibration methods that might facilitate the implementation of spirometry in nonspecialized settings. This project will be conducted in the occupational cohort followed at Mount Sinai, among the WTC Pulmonary Evaluation Unit Chest CT Imaging Archive subcohort members still on active health surveillance. This group of 1710 WTC workers has detailed disease symptom, both pre-WTC and WTC-related occupational exposure, longitudinal spirometry measurements and body weight trends, visual chest CT imaging classification and grading, and quantitative computer tomography (QCT) measurements of airway, body composition, and pulmonary parenchymal and vascular abnormalities.

NIH Research Projects · FY 2024 · 2024-07

PROJECT SUMMARY: In the subsequent years following the World Trade Center (WTC) attack on September 11th, 2001, a cluster of chronic health conditions emerged among first responders (FRs) who were present during the aftermath of the attack. Many FRs were at Ground Zero for prolonged periods in the first week after the disaster, repeatedly exposing them to high levels of dust composed of particles of varying sizes that contained metals, polychlorinated biphenyls, polyaromatic hydrocarbons, among many known toxins. A growing body of scientific literature indicates that FRs who were exposed to high levels of dust for prolonged periods may have a greater incidence of mild cognitive impairment (MCI), as well as other neurological complications i.e. changes in white matter connectivity, decreased hippocampal volume which may put them at a greater risk of developing Alzheimer's disease (AD). Crucially, the risk relating to exposure was elevated in individuals carrying the AD vulnerability gene (i.e., apolipoprotein-E4 [APOE4], TREM2). Carrying a single APOE4 allele increases the risk for AD by approximately 4-fold, while double carriers have a 10-fold increased risk compared to the neutral APOE3 isoform. Based on this consideration, we have established the Late onset of Alzheimer's disease (LOAD)2 mouse model which carries a humanized ApoE knock-in mutation for the E4 isoform, a CRISPR/Cas9- generated APP allele with a humanized Aß1-42 region and R47H point mutation in the TREM2 gene to study the effects of WTC airborne PM and the role of age and genetic and environmental risk factors on LOAD. Additionally, in new studies, we found that the combination of experimental “extreme weather conditions” by heat stress (HS) in mice, primed by exposure to WTC airborne PM, exacerbates impairment in energy metabolism, altered lipid metabolism, and inflammatory responses. Consistent with this hypothesis, our feasibility evidence shows that the combination of environmental risk factors (such as HS) to a single WTC airborne PM exposure may negatively influence several metabolic cascades associated with energy metabolism. Based on this, our proposed study will define an age-dependent interaction regarding environmental “climate changes” (HS) to WTC airborne PM exposure on the onset and progression of LOAD. The overarching goal of this proposal is to examine the impact of WTC airborne PM on age x genetic interactions in response to exposure to WTC airborne PM and/or extreme weather conditions (HS), ultimately providing new strategies for risk assessment, risk reduction, and disease prevention in LOAD. The choice to study the role of WTC airborne PM in LOAD2 mice is fundamental to our understanding of age x genetic interactions in response to exposure to environmental stressors. The proposed studies seek to explicitly test the hypothesis that exposure to toxic WTC airborne PM causes long-term cognitive impairment and AD-type pathology by using rodent models. We understand and appreciate the importance of sex as a biological variable. However, for rigor and reproducibility purposes, we would like to note that this study is not designed to specifically analyze sex differences.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Many patients who have recovered from SARS-CoV-2, the virus that causes COVID-19, continue to experience a constellation of symptoms long after the initial illness. Known as “long-COVID”, or Post- Acute Sequelae of SARS-Cov-2 infection (PASC), the most frequently reported symptoms are fatigue, post exertional malaise and cognitive dysfunction, which are also the primary symptoms of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS). Many of the PASC patients fulfill diagnostic criteria for ME/CFS, but differ from non-PASC ME/CFS patients in that they share a common infectious trigger and have a shorter duration of illness, which reduces heterogeneity. Understanding whether PASC ME/CFS shares overlapping mechanisms with non-PASC ME/CFS is critical, as this could provide insights into the mechanisms and inform treatment strategies of ME/CFS in general. To address this question, we propose a comparison study of PASC ME/CFS patients with sudden onset illness to non-PASC ME/CFS patients who reported a sudden flu-like illness onset. Limited studies have shown reductions in cerebral blood flow and increased cerebroventricular lactate in ME/CFS patients suggesting alterations in perfusion and metabolic properties. Our recent preliminary results show that the oxygen extraction fraction was elevated in PASC ME/CFS patients, which may be attributed to reduced cerebral blood flow and mitochondrial dysfunction. In this project, we aim to conduct non- invasive brain magnetic resonance imaging (MRI) to compare the similarities and differences in cerebral oxygen and glucose metabolism between the two patient groups as well as healthy controls. We will measure and compare the oxygen extraction fraction, cerebral blood flow, and cerebral metabolic rate of oxygen and glucose uptake and metabolic rate in the patient groups and healthy controls. The MRI derived parameters will then be correlated to the disease symptom burden. Additional, since many PASC patients recover over one year, we aim to perform a follow-up study on the PASC and non-PASC ME/CFS groups. Completion of this timely and important study will provide comparison of PASC and non-PASC ME/CSF in terms of changes in glucose and oxygen metabolic properties, as well as how these imaging parameters are related to the disease burden. Through analysis of the longitudinal data, we will be able to determine whether the changes in metabolic properties are associated with changes of patient reported outcome measures. The knowledge learned will deepen our understanding of the ME/CFS/PASC (long-COVID) disease mechanisms, aid in ME/CFS diagnosis, inform treatment decisions, and inspire new treatment targets.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Parkinson’s disease (PD) is a devastating neurodegenerative disease characterized by a loss of dopaminergic neurons in the substantia nigra, movement abnormalities, and in some cases the accumulation of α-synuclein (α-syn) as fibrils or Lewy bodies. It is well established that T-cells are involved in PD pathology, shaping the inflammatory cascade from the periphery into the brain, and should be the focus of additional studies. Immune alterations in PD are found in peripheral blood indicating a pro-inflammatory status during disease and as early as prodromal stages. However, specific alterations are disputed and must be clarified with a large-scale study and a carefully curated single-cell cohort that controls for biological heterogeneity. T-cell infiltration into the brain has been reported to parallel α-syn accumulation and neuronal death but exploring infiltrated T-cells to characterize the role of these cells in neurodegeneration is a challenge. Genome-wide association studies (GWAS) have identified human leukocyte antigen (HLA) class II PD risk loci, which suggests an unexplored important role for CD4 T-cells in their interaction with antigen presenting cells. Overall, this work evaluates T-cell biomarker potential by investigating their contribution to PD in different clinical phases and explores their mechanistic role in pathogenesis. In this proposed work, the role of T-cells, particularly antigen-specific ones, in PD pathogenesis will be explored by characterizing immune cell expression and repertoire in PD patients, prodromal patients (who have elevated risk of disease, including GBA/LRRK2 genetic carriers and patients with rapid eye movement (REM) sleep behavior disorder) and control donors. Further, the clinical relevance and antigen specificity of clonal T- cells will be determined by comparing with α-syn-specific cells identified in in-vitro stimulation experiments. Investigating prodromal patients with an elevated risk of developing PD will assess the importance of T-cells in disease development and their potential as biomarkers. Additionally, harnessing the human Living Brain Cohort data, I will compare gene expression and immune repertoire between circulating and infiltrated T-cells from the same donors at a single-cell level. Comparing the T-cell receptors (TCRs) in the blood and brain provides a critical link between T-cells and disease progression, and can reveal whether T-cells of particular specificities have infiltrated into the brain parenchyma. This work will also substantiate the relationship between risk/protective HLA alleles and T-cell activity in PD patients by investigating the association between HLA alleles and TCR gene usage and the hypervariable complementarity determining region 3. The results of these experiments will further our comprehension of the role of T-cells in PD, lead to mechanistic discovery and determine the potential for targeted immune-based therapeutic interventions or biomarker capabilities.

NSF Awards · FY 2024 · 2024-07

The broader impact of this I-Corps project is based on the development of an improved approach to treating Glioblastoma to improve patient outcomes and well-being. By offering a minimally invasive, potentially more effective treatment, this approach is expected to result in improved patient outcomes and quality of life, addressing a critical need within oncology care. The innovation underlying the solution enriches magnetic hyperthermia therapy (MHT) and nanoparticle technology. This project illustrates the potential for cutting-edge treatments to influence healthcare policies towards more innovative and cost-effective solutions, offering economic benefits and setting a precedent for future medical technologies. Furthermore, this project underscores the societal value of investing in health innovation for the betterment of patient care and treatment possibilities across various diseases. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of the technology. The solution is based on the development of an innovative treatment system for Glioblastoma, a highly aggressive brain cancer with limited current treatment options. This system utilizes Magnetic Hyperthermia Therapy (MHT) with magnetic iron oxide nanoparticles to selectively target and destroy tumor cells while preserving surrounding healthy tissue. This solution integrates multiple functionalities into a single, minimally invasive device, including real-time temperature monitoring for precise therapy delivery, tumor debulking, biopsy capabilities, and electrocautery to minimize blood loss. This approach has shown promising results in small animal trials, indicating its potential to significantly improve Glioblastoma treatment outcomes. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract By 2030, the majority of World Trade Center (WTC) rescue and recovery workers (responders) will be aged 65 and over and at risk for aging-related conditions and consequences. Because the WTC General Responder Cohort (WTC-GRC) is aging (median age is 61 in 2023), the WTC Health Program (WTCHP) needs to be prepared to deliver care that is best aligned to the needs of the aging WTC responder population. Frailty, a common geriatric syndrome characterized by increased vulnerability to adverse events including mortality, morbidity, disability and hospitalizations is prevalent among the WTC-GRC population and will likely continue to grow as the population ages. Our team has previously demonstrated that approximately 30% of the WTC- GRC meets criteria for frailty, an association that increases with age, WTC exposure, and by occupation type (e.g., higher risk among construction worker). These findings underscore an urgent need for the targeting of potential vulnerabilities of this population as they age. This project will investigate two key questions central to developing aging-related interventions in the WTCHP. The first question is to understand how evidence-based healthy aging interventions can be best implemented in the WTCHP to meet the needs of WTC responders. This study will examine the implementation of a deprescribing intervention as an evidence-based practice to improve the use of medications in aging populations. Key information about implementation of this practice in the Mount Sinai WTCHP will be examined to identify barriers to implementation and identify strategies, processes and tools to support its implementation. Perspectives from key stakeholders at a novel site for the evidence-based practice will be collected in order to prepare for further dissemination of this deprescribing intervention. The second question is to understand health priorities of the WTC-GRC as they age. Examination of these health priorities will allow for further planning of healthy aging initiatives, as well as selection and prioritization of additional evidence-based practices needed in future efforts to support WTC responders. Results of this project will be instrumental in improving the capacity of WTCHP to care for aging responders through partnership with Mount Sinai's nationally recognized Geriatrics and Palliative Care programs including the development of future widespread multifactorial programs tailored to WTC responders to promote healthy aging.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Podocyte abnormalities have been identified in many kidney diseases. The structural role of podocytes in the filtration barrier is known to be a dynamic process in which the podocyte actively restructures its actin cytoskeleton. Mechanical stress, such as severe hypertension, can have a detrimental impact on this process. LIM-nebulette is an actin binding protein within podocytes. We hypothesize that this protein helps podocytes adapt to the mechanical microenvironment in vivo. We have engineered an instrument with deformable surfaces to simulate the mechanical tensile forces experienced by podocytes in vivo. The goal of this project is to combine high-content image analysis with a novel mechanical stretch platform to understand how podocytes maintain their structural integrity against injury and use this system to study the operation of an important cell signaling pathway in the glomerular environment. Stretched and control samples will also be collected for fluorescent microscopy, protein expression, phosphproteomics and co-immunopreciptation to determine nebulette’s signaling network. Understanding the role of LIM-nebulette in maintaining podocyte cytoskeletal integrity could potentially reveal new avenues for stabilizing these cells and, consequently, preserving kidney function. By compiling and analyzing this data, we aim to clarify the role of LIM-nebulette in protecting podocytes. This project will yield cellular level insights into podocyte activity under glomerular environmental stress conditions by examining morphometric changes and alterations in mechanobiological signaling that may have an impact on chronic kidney disease.

NSF Awards · FY 2024 · 2024-07

The broader impact of this I-Corps project is based on the development of an intracranial device to treat neurodegenerative disease, such as Alzheimer’s or Parkinson’s. Current treatment devices involve helmets or headwear, and stigma around these bulky devices affects patient self-esteem and deters patient usage. This technology aims to be mostly intracranial (with the potential exception of small, flush hardware extracranially), and thus, may be associated with better patient satisfaction and well-being. Furthermore, given the estimated $655 billion spent yearly on neurodegenerative diseases in the United States, this technology has potential for financial impact if successful in slowing progression or alleviating symptoms of neurodegenerative diseases. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of the technology. The solution is based on the development of an intracranial device that works through cellular photobiomodulation (PBM) to treat neurodegenerative diseases, such as Alzheimer’s or Parkinson’s. This approach creates minimally invasive burr holes over affected regions of the brain, which would be closed with a direct targeting photobiomodulation device to apply the therapy uncompromised by attenuating obstructions. For example, PBM holds promise as a postsurgical healing tool, especially in surgeries that require burr holes, by eliminating the need to penetrate additional tissue to reach clinically relevant structures. This device could deliver therapeutic benefit for certain neurological afflictions and could contribute to a better understanding of the mechanisms of PBM therapy that may inform targets for novel therapies. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

The overarching objective of this KPMP Central Hub application is to create and support a collaborative environment committed to promoting participant safety, scientific rigor, patient engagement, high ethical standards, and the interdisciplinary team science necessary to result in major advances in kidney precision medicine. The Central Hub will coordinate the KPMP Recruitment Sites, Tissue Interrogation Sites, and Opportunity Pool research groups; engage the broader patient, clinical, and research communities; and partner with the Kidney Tissue Atlas Coordinating Center to securely, efficiently, and transparently deliver Findable, Accessible, Interoperable, and Reusable data to these communities. Our experienced multidisciplinary team was instrumental in building the infrastructure and functional ecosystem needed to launch and maintain the first phase of the KPMP. We will build on this experience to provide scientific, technical, administrative, operational, and logistical support to pursue the following specific aims: (1) coordinate ethical, safe, and rigorous implementation of KPMP protocols and procedures; (2) support the collection and distribution of high-quality data and samples with rigorous quality assurance and quality control; (3) build, support, and maintain a collaborative culture within KPMP that continuously honors and values the contributions of study participants; and (4) foster external collaborations and catalyze new partnerships through an Opportunity Pool of funds dedicated to maintaining KPMP dynamism.

NIH Research Projects · FY 2025 · 2024-06

Abstract Respiratory viruses spread from host to host as they are shed from mucosal surfaces and can transmit through the air or via surface contact. Therefore, it is believed that the induction of a strong mucosal immune defense cannot only prevent from infection at the port of virus entry, but also affect virus transmission, thereby preventing new infections. In this proposal, we will address the impact of mucosal adjuvanted vaccination on the outcome of disease, host immune responses to and transmission events after experimental respiratory infection with influenza virus and SARS-CoV-2. We will use the guinea pig (influenza) and Syrian golden hamster (SARS-CoV- 2), two established models for vaccination, infection and transmission studies with respiratory viruses. Intranasal virus inoculation will result in replication mainly in the upper respiratory tract for guinea pigs, whereas SARS- CoV-2 will replicate in both upper and lower respiratory tract. We will compare protection provided by mucosal adjuvanted vaccination via the intranasal route to parenteral vaccination via the intramuscular route and will investigate the impact of induction of protective immune responses at mucosal sites on host immune responses to infection in vaccinated hosts (hybrid immunity) and virus transmission between vaccinated infected animals and naïve receiver animals. Despite being vaccine-preventable diseases, transmission of influenza and SARS- CoV-2 occurs frequently after breakthrough infection of antigenically drifted virus variants in vaccinated people. For this R21, three principal investigators with complementary expertise in preclinical animal models for respiratory virus transmission, mucosal adjuvanted vaccines and immune profiling will continue to build on their previous collaborative efforts and expertise to investigate the potential of an intranasal mucosal adjuvanted vaccine strategy previously validated in mice. The mucosal adjuvant consists of a nanoemulsion with TLR- agonistic properties combined with a RIG-I agonist. This combination adjuvant is currently being tested and compared to other licensed intramuscular adjuvants by us in mice for its suitability as a mucosal adjuvanted vaccination strategy for universal and supraseasonal influenza and SARS-CoV-2 vaccine antigens. The efficiency of inhibition of transmission by mucosal vaccination will be compared to parenteral vaccination with the current standards of care for populations at risk (split inactivated influenza virus vaccine or recombinant hemagglutinin for influenza and mRNA or adjuvanted recombinant SARS-CoV-2 spike protein-based COVID-19 vaccines). We will correlate induced mucosal and systemic immune responses after vaccination with protection from experimental infection, reduction in transmission to naive receiver animals and enhancement of hybrid immunity (immunity as a result of vaccination and infection).

NIH Research Projects · FY 2026 · 2024-06

Late onset Alzheimer’s disease (AD), the most common form of dementia, is characterized by memory loss and a progressive decline in cognitive function. The Accelerating Medicines Partnership-Alzheimer’s Disease (AMP- AD) program has supported systems biology analyses, with VGF (non-acronymic), a neurotrophin-inducible protein and neuropeptide precursor, being one of the top ranked AD drivers (regulators) identified by several groups. Notably, biomarker studies have consistently identified reduced VGF levels in the brains and CSF of patients with AD, and have further demonstrated that VGF is also a strong candidate biomarker of AD progression. Our recent published data demonstrate that VGF overexpression in hippocampus or chronic intracerebroventricular (icv) infusion of the VGF-derived neuropeptides TLQP-21 or TLQP-62 (named by the N- terminal 4 amino acids and length) reduces cortical and hippocampal amyloid deposition, microgliosis, and/or astrogliosis, in a regionally-specific manner, and reduces cognitive impairment in the 5xFAD mouse amyloidosis model. TLQP-21 activates the complement C3aR1 G-protein coupled receptor (GPCR), also implicated in AD pathogenesis and expressed in the CNS on neurons, microglia, and astrocytes, while the antidepressant and procognitive functions of TLQP-62 are dependent on BDNF/TrkB signaling. In this RO1 proposal, we request support for preclinical studies that will optimize intranasal (IN) nose-to-brain therapeutic delivery of the VGF- derived peptides TLQP-21 and TLQP-62 to mice, and will determine their efficacy in the 5xFAD mouse model of amyloidosis and then subsequently in the PS19 tauopathy model to reduce neuropathology and the progression of AD-like phenotypes. Our preliminary imaging data indicates that intranasal administration of Cy5-TLQP-21 and Cy5-TLQP-62 results in significant, rapid brain uptake. Aim 1 will determine regional uptake in the brain and any systemic spillover of IN Cy5-conjugated peptides, using small animal imaging and western analysis of brain and peripheral tissues. Aim 2 will verify retention of intact peptide in brain using Cy5- and biotin-conjugated peptide quantification by western analysis, and will determine ‘target engagement’ in hippocampus and cerebral cortex by IN peptide, including BDNF/TrkB/CREB and c-fos activation. Interim milestones include identification of the optimal dose and dosing interval for IN peptide delivery based on quantification of regional peptide levels in the brain, and the determination that signaling pathways known to be regulated by intra-hippocampal or icv delivery of these peptides are activated by IN peptide. Aim 3 will utilize the most efficacious IN delivery schedule identified to determine whether cognitive impairment and neuropathology in 5xFAD and PS19 can be reduced. Lastly, longevity of intranasal peptide treatment effects will be tested, systemic toxicity will be assessed, peptide modification(s) will be considered, and behavioral, biochemical, histological and large-scale genetics approaches will be employed to measure relative peptide efficacy in reducing disease progression. These preclinical studies will provide a firm foundation for future intervention using intranasal delivery of VGF peptide therapeutics in AD.