University Of Pennsylvania

NSF Awards · FY 2024 · 2024-09

This project studies interacting particle systems at the interface of probability, combinatorics, statistics and applied mathematics. Interacting particle systems naturally emerge in various physical systems, and their behaviors—such as global fluctuation and relaxation rate— are crucial in understanding these systems. Beyond pure mathematics, interacting particle systems have been widely used for random sampling, due to their flexibility and high accuracy. This project will develop new tools and techniques for advancing the study of interacting particle systems. The project will also provide valuable educational opportunities for students at several levels, including a summer school and workshops focusing on the interactions of probability, mathematical physics, and machine learning theory. These initiatives seek to bring together early-career researchers from diverse fields to foster collaborative efforts. One primary objective of this project is to investigate the asymptotic behaviors of various interacting particle systems, such as local statistics, global fluctuations and large deviations. These findings will facilitate deriving asymptotic properties of symmetric polynomials, a task less accessible through traditional algebraic combinatorics methods. Additionally, new universal laws, such as the Tracy-Widom distribution, appear in both random matrix theory and intersecting particle systems. The scaling limits of interacting particle systems converge to two-dimensional universal limiting objects, from which we can recover random matrix eigenvalue statistics by taking one-dimensional slices. Another key aim is to employ tools from interacting particle systems to establish characterizations of random matrix statistics, particularly from the view of line ensembles. These characterizations provide novel pathways to understand convergence to random matrix statistics in many statistical physics models. Lastly, the PI aims to develop efficient particle-based derivative-free sampling algorithms. These algorithms will enable effective uncertainty quantification for models where differentiation is impractical. 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 · 2024-09

Summary We propose to create a comprehensive training program that encompasses research training in science related to kidney, benign urology, and benign hematology across the lifespan entitled the Philadelphia Program for Mentored Research Training in Kidney, Urologic, and Hematologic Diseases (PERFORM-KUH). Leveraging the physical proximity and the history of interinstitutional and interdisciplinary training and collaboration, PERFORMKUH is uniquely positioned to build/enhance a biomedical research training program that integrates well-established training programs at the University of Pennsylvania (Penn) and the Children’s Hospital of Philadelphia (CHOP), and pre- and post-doctoral programs at Drexel, Jefferson, and Temple (with affiliated Fox Chase Cancer Center) Universities. The primary goal of this program is to develop skilled investigators trained to conduct impactful research in clinical, translational, or basic science in KUH, and capable of further developing careers as independent researchers. PERFORM-KUH will replace three longstanding T32 training programs located at Penn/CHOP, in hematopoiesis, kidney disease, and renal epidemiology, add training in pediatric and adult urology research, and importantly, integrate with three partner institutions in Philadelphia. This program assembles one hundred and six talented faculty trainers from five institutions, sixteen graduate programs, and thirty-eight departments, to create a premiere integrated research training program for KUH trainees in the Philadelphia region. PERFORM-KUH is designed to support eight predoctoral and twelve postdoctoral trainees per year. This request is justified by the historical demand by high-quality applicants for training slots in our standing T32s, accommodated by the size and quality of the PERFORM-KUH faculty, large and outstanding trainee pools, strong institutional and departmental commitments, and collaborative training environment. The PI/PD with focus area advisors and site PIs are well supported by the Steering Committee of the Administrative Core consisting of a Recruitment and Admissions committee, Mentorship Oversight subcommittee, and an External Evaluation Board. Research mentorship is provided by scientifically diverse trainers of all academic ranks with research interests that encompass virtually all areas of KUH. The Training Program will leverage innovative PERFORM-KUH cores, including its Professional Development Core (PDC), and its Networking Core (NWC) that integrate pipeline programs to attract undergraduate, medical, and high school students from underrepresented backgrounds. The DEI director is dedicated to minority recruitment and retention efforts in PERFORM-KUH research training. The academic elements of the Program include laboratory work, skills training, seminars, mentoring and career counseling, presentation, manuscript and grant writing, and peer mentoring in collaboration with PDC and NWC. It will offer an integrated core curriculum that includes the completion of a formal master’s degree program for MD research fellows. Facilitated by a professional Evaluation Director, PERFORM-KUH will be evaluated using systematically collected and formally analyzed data. Project Summary/Abstract

NIH Research Projects · FY 2024 · 2024-09

SUMMARY This application requests funding to support the 2025 International Workshop on Pulmonary Imaging, a three- day meeting to be held from February 20-22, 2025, and the eighth iteration of this workshop to take place at the University of Pennsylvania. The majority of requested funds will be used to cover the cost of travel and lodging for junior researchers—undergraduate and graduate students, as well as postdoctoral fellows and junior faculty—presenting at the workshop. A smaller portions of these funds will be allocated to help pay for publication costs related to workshop materials as well as live webcasting so that interested individuals around the world who are unable to attend in-person still have the opportunity to view and participate in the proceedings. As pulmonary disease represents the third leading cause of mortality worldwide (rising to almost 4 million deaths annually), there is an increasing need for novel imaging techniques to provide earlier/more accurate diagnoses and improved treatment monitoring. The field of pulmonary imaging is more expansive today than ever— spanning an unprecedented range of techniques for structural, functional and molecular lung assessments being developed across the disciplines of physics, biology, engineering, chemistry, computer science, medicine and more. Given its rapidly-evolving nature, the existence of a regular forum in which scientists and clinicians in the field can meet with and communicate their ideas to one another is of the utmost importance. In the absence of other scientific meetings with a similarly focused agenda, our previous workshops have succeeded in providing just such a forum—creating a valuable opportunity for rigorous, collaborative exchange. The specific aims of the proposed workshop are as follows: (1) host a one-day workshop on pulmonary fibrosis; (2) keep the pulmonary imaging community informed regarding the most recent developments in structural, functional and molecular lung imaging; (3) investigate the use pulmonary imaging to non-invasively assess treatment efficacy for interstitial lung diseases (ILDs); (4) explore novel approaches for integrating deep learning with pulmonary imaging to improve diagnosis and phenotyping and predict disease progression; (5) broadcast the full workshop program live online, allowing interested parties free, real-time access to the proceedings. Finally, we intend to hold a one-day boot camp the day before the conference focused on pulmonary fibrosis. This boot camp will consist of ~5 one-hour presentations on topics related to this central theme, with significant question and answer time, and a final discussion panel during which the goal will be to synthesize the most important issues covered over the course of the event. Finally, the 2025 workshop will expand its focus to include new dedicated sessions on topics such as pulmonary vascularization, small airways disease, ILDs—and specifically idioopathic pulmonary fibrosis (IPF)—while also the maintaining recent emphasis on assessing treatment response through imaging, the integration of deep / machine learning techniques, and bridging the gap between current clinical practice and innovative imaging technologies.

NIH Research Projects · FY 2025 · 2024-09

Females with sickle cell disease (SCD) in the United States face a health disparity triple threat as a result of their chronic disease, race, and sex. In addition to overall reduced survival rates in both sexes and maternal mortality rates that exceed those of Black females without SCD, females with SCD experience significantly more frequent and more severe vaso-occlusive pain episodes (VOEs) compared to their male counterparts. However, the pathophysiology of this pain disparity – whether biological, sociocultural, or both – is not understood. With this proposal, we will test our central hypothesis, that female sex hormones modulate cyclic increases in inflammation that predispose to VOEs. SCD is characterized by chronic inflammation, with acute rises in inflammatory markers during VOEs. VOEs exhibit a perimenstrual pattern, peaking during or prior to menses in many females with SCD. Our preliminary data show that inflammatory markers may exhibit a similar pattern, peaking in the follicular phase. Together, the established correlation between pain and inflammation and newer evidence of a cyclic pattern to pain and inflammation in SCD lead us to postulate that female sex hormones mediate inflammation and VOEs. Moreover, hormonal contraception may play a therapeutic role in SCD. Depot medroxyprogesterone (DMPA, Depo-Provera®) is a progestin-only contraceptive that reduced VOE frequency in small studies. Since DMPA suppresses hormonal fluctuations in the menstrual cycle, we hypothesize that DMPA may reduce cyclic fluctuation in inflammation and thereby relieve VOE pain. The pathophysiologic pathway of perimenstrual VOE is likely multifactorial. The objectives of this study are (1) to ascertain sex-related mechanisms of perimenstrual VOE and (2) to determine the therapeutic effectiveness of DMPA on VOE and inflammatory biomarkers. We propose two prospective studies of patients with SCD recruited from university-based clinical practices at the University of Pennsylvania and Emory University. In Aim 1, “Inflammatory Biomarker Patterns in Females and Males with Sickle Cell Disease,” we will evaluate traditional and functional inflammatory biomarkers via serial blood sample collections across the menstrual cycle in 20 females with SCD, and compare to serial biomarker levels over one month in 8 male controls with SCD. In Aim 2, “Clinical and Biomarker Effects of Depot Medroxyprogesterone Acetate in Females with Sickle Cell Disease,” we will assess 52 females with SCD pre- and post-intervention with DMPA to measure effects on VOEs and the same biomarkers. With these studies, we plan (1) to demonstrate that key inflammatory biomarkers are modulated by sex and (2) to determine whether DMPA prevents VOEs and reduces inflammation. This research will form the scientific basis for new horizons for VOE prevention in females with SCD.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT As large-scale genome-wide association studies (GWAS) continue to yield now thousands of genomic loci robustly associated with neurodevelopmental and psychiatric disorders, including autism spectrum disorder (ASD) and schizophrenia (SCZ), the major defining challenge of the post-GWAS era is to characterize the concrete biological mechanisms through which this polygenic variation confers disease risk, at scale. To this end, we and others have recently developed methods and resources for systematic integration of GWAS results with population-level functional genomic reference panels -- identifying isoform-regulation during the second trimester of human brain development as mediating the greatest proportion of heritability across multiple neuropsychiatric GWAS studies compared with earlier or postnatal timepoints. Yet, no studies have characterized genetic regulation of alternative polyadenylation (APA) in the developing brain, a critical yet understudied tissue-specific gene-regulatory mechanism with established roles in neuronal mRNA metabolism, subcellular trafficking, and cellular differentiation. Our preliminary data indicates widespread dysregulation of APA in stem-cell-based models and postmortem brain tissues from subjects with ASD and SCZ, as well an outsized enrichment of psychiatric GWAS signal with APA quantitative trait loci (QTL) in the developing human brain. This proposal seeks to integrate large-scale functional genomics, single-cell and long-read sequencing, deep learning, and genome-editing in human neuronal stem-cell models to develop a detailed, mechanistic understanding of APA regulation during human brain development and its contribution to neuropsychiatric disorder pathophysiology. Specifically, we will generate a comprehensive atlas of APA regulation across neurodevelopment, leveraging data from >3650 bulk tissue samples as well as single-nucleus RNAseq data across >700 unique donors, including 170 with SCZ/ASD. We will train and validate a deep learning model predicting APA changes from primary sequence. Through integration with psychiatric GWAS, we hypothesize that APA regulation will provide substantially greater resolution to detect candidate biological mechanisms underlying psychiatric GWAS loci. Finally, predicted SNP-UTR-disease mechanisms will be experimentally tested via high-throughput screens and genome-engineering in iPSC-derived neurons. Together, these studies will systematically characterize a critical, yet underexplored area of genomic regulation in the human brain across development, thereby providing novel insights into psychiatric disease mechanisms and identifying potential neurobiological targets for therapeutic development and intervention.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The expression levels of protein arginine methyltransferases (PRMTs) are significantly elevated in cancer cells and are associated with poor clinical outcomes. This strongly suggests their potential as a novel class of therapeutic targets in oncology. Major pharmaceutical companies have developed over ten selective and potent PRMT inhibitors (PRMTis) that are currently in early-stage clinical evaluation. However, due to the understudied nature of arginine methylation as a post-translational modification, a comprehensive mechanistic understanding of PRMT functions in cancer is urgently needed to enable the clinical application of these inhibitors. Notably, recent clinical trial results indicate that the activity of PRMTis alone may be insufficient for effective cancer management in patients. Therefore, it is imperative to conduct mechanism-driven preclinical evaluations of combination strategies that target PRMTs alongside other therapeutic drugs to achieve clinical success. Both high-grade serous ovarian cancer (HGSOC) and triple-negative breast cancer (TNBC) share clinical and genomic characteristics such as poor prognosis, homologous recombination deficiency, and potential immunoreactivity. Given the urgent unmet medical need to develop effective therapeutic strategies for these diseases, our preliminary studies have merged large-scale drug combination screening and cancer genomic profiling to establish a strong rationale for applying PRMTis in treating HGSOC and TNBC. Our hypothesis is that because of uncontrolled proliferation, increased DNA damage, and activated oncogenes, cancer cells must sustain abnormal levels of transcription, splicing, and protein arginine methylation through enhanced PRMT activities (PRMT addiction). Treatment with PRMTis can disrupt the hyperactivated PRMT-regulome that enables cancer cells to survive. We've assembled a team of investigators with diverse expertise and resources to test this hypothesis through three specific aims: Specific Aim 1: Characterize molecular mechanisms of PRMTi mono- and combination therapies; Specific Aim 2: Evaluate therapeutic potentials of PRMTis in HGSOC and TNBC preclinical models; and Specific Aim 3: Investigate effects of PRMTi treatments on anti-tumor immune responses. The overarching objective of this application is to systematically explore the dynamic changes in the regulome induced by PRMTi treatments in cancer cells. This will offer fresh insights into their mechanisms of action and provide a rationale for the clinical development of PRMTi mono- and combination therapies in the field of oncology.

NIH Research Projects · FY 2025 · 2024-08

The Surviving Sepsis Campaign emphasizes the critical need to provide sepsis survivors with patient education tailored to their self-care management after hospital discharge. This includes education on their diagnosis, treatment, infection prevention, early recognition, timely treatment of infection, and post-sepsis syndrome management. Many sepsis survivors are unaware of their diagnosis or its implications for self-care after discharge, leading to knowledge gaps that contribute to delayed follow-up care, poor health outcomes, and frequent rehospitalizations. Sepsis survivors often face long-term morbidity symptoms and are twice as likely to be rehospitalized within 30 days of discharge compared to the general inpatient population. Most of these rehospitalizations are due to new or recurring sepsis or infection and are considered preventable through early post-discharge self-care. Home health care (HHC) offers a key opportunity for delivering sepsis patient education, as 13.1% of Medicare sepsis survivors are discharged to HHC. According to the U.S. Centers for Medicare and Medicaid, patient education is a vital part of HHC, and preliminary findings from a qualitative study suggest that acute care clinicians believe sepsis patient education would be better facilitated within HHC rather than in the hospital setting. However, sepsis survivors discharged to HHC experience higher 30-day rehospitalization rates than those discharged to skilled nursing facilities. Since HHC does not provide continuous patient monitoring, high-quality and personalized patient education is essential to help survivors recognize symptoms and seek timely care. Despite this, little is known about sepsis patient education within HHC and how prepared survivors feel to manage their recovery. This qualitative descriptive study will generate foundational knowledge on sepsis patient education in HHC to inform future interventions aimed at improving self-care among sepsis survivors. The specific aims are: 1) to characterize the self-care self-efficacy and knowledge regarding sepsis and its aftercare among recent hospital discharges receiving HHC; 2) to identify the materials, content, mode of delivery, facilitators, barriers, and strategies used by HHC nurses in delivering sepsis patient education; and 3) to describe the barriers and facilitators influencing the receipt and understanding of sepsis aftercare among survivors within HHC. This fellowship provides opportunities to enhance knowledge of current patient education interventions, develop expertise on sepsis survivor health outcomes, apply rigorous qualitative methods, and advance my independent research program. This study addresses the NINR research lens on Systems and Models of Care, which “focuses on how healthcare and health services are delivered.”

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Recent developments in spatial transcriptomics (ST) technologies have enabled the comprehensive profiling of gene expression across tissues while preserving crucial gene location information. However, the current accessibility of commercialized ST platforms remains constrained by their high costs and long turn-around time, limiting their utility in large-scale ST studies that involve hundreds or thousands of samples. In contrast, hematoxylin and eosin (H&E)-stained histology images are much cheaper to generate. Previous studies have demonstrated correlations between gene expression patterns and histological image features, suggesting the potential to predict spatial gene expression from histology images. Therefore, the integration of histology image data for predicting spatial gene expression in large-scale ST studies has emerged as a promising strategy. This novel approach facilitates the generation of virtual ST data at significantly reduced costs and time commitment. Through these predictive models, we can investigate the intricate connections between spatial gene expression variations and clinical outcomes of interest, with a particular focus on population-based inquiries, such as those in biobank samples. Leveraging our team's expertise in ST data analysis and computational pathology, we propose to develop a suite of informatics tools to harmonize information from histology images and integrate it with ST data with diverse spatial resolutions and gene coverages to build gene expression prediction models. We will further investigate optimal sample selection strategies for ST when conducting a large-scale ST study. These tools will make it possible to predict ST data at single-cell resolution from samples where only histology images are available. To enhance the impact of our research, we will also develop open-source software packages and build a cloud-based computing platform for ST data prediction and visualization in large-scale studies. Realizing the proposed research would signify a transformative advancement in the field, potentially leading to a paradigm shift.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY βII-spectrin, a ubiquitous component of the neuronal cytoskeleton, plays multifaceted roles in the organization and function of neurons. We recently reported that de novo variants in SPTBN1, which encodes βII-spectrin, cause a pediatric syndrome characterized by global developmental delay, intellectual disability, ADHD, autism spectrum disorder (ASD), movement deficits, and epilepsy. However, the neuronal types and pathways that are most vulnerable to deficits in βII-spectrin function are largely unexplored. We found that mice with selective loss of βII-spectrin in cerebellar granule cells exhibit symptoms and behaviors consistent with the clinical presentations observed in individuals carrying pathogenic SPTBN1 variants. The cerebellum, previously assumed to mostly modulate fine-motor coordination, plays important roles in cognition and is suspected to contribute to the pathophysiology of neurodevelopmental diseases. Thus, it is important to define the normal and pathogenic mechanisms modulated by βII-spectrin function in the cerebellum. To accomplish these goals, we will combine cellular, biochemical, live and super-resolution microscopies, and electrophysiological techniques with behavioral paradigms to 1) determine the molecular mechanisms underlying disruption of excitable axonal domains and synaptic transmission in cerebellar granule cells caused by βII- spectrin deficiencies and 2) leverage novel transgenic mouse models carrying clinically relevant Sptbn1 variants to assess their effect on cerebellar molecular, cellular, and synaptic function and clinically relevant behaviors.

NIH Research Projects · FY 2025 · 2024-08

Increasing efforts are being made to understand whether early intervention after HIV-1 infection can enable long- term viral control and reduction of the viral reservoir through the preservation of HIV-1-specific immune responses. Recently, the pioneering eCLEAR phase 1b/2a clinical trial led by MPI SØgaard, found that administration of the HIV-Env specific broadly neutralizing antibody (bNAb) 3BNC117 at antiviral therapy (ART) initiation enhanced CD8+ T cell viral immunity and enabled viral control amongst participants harboring 3BNC117-sensitive viruses. However, the mechanisms underlying this outcome remain largely unknown. Understanding The goal of this collaborative R01 (M Betts, UPenn, B Jones, Weill-Cornell University, and O. SØgaard, Aarhus University) is to define the mechanisms underlying this protective outcome in order to inform future development of targeted immunotherapy at ART initiation as an HIV cure strategy. We hypothesize that the partial success of eCLEAR can be defined in terms of both specific mechanisms of efficacious CD8+ T-cell responses and of the features selected in remaining reservoir-harboring cells. We will address this hypothesis in three integrated Aims using samples directly from the eCLEAR study. In Aim 1, we will define phenotypic, functional, and transcriptomic features of the adaptive T cell immune responses that are modulated by bNAb treatment at ART initiation (SØgaard). In Aim 2, we will define how the HIV reservoir is differentially modulated after prolonged ART treatment and within post-therapy viral controllers vs. non- controllers who received 3BNC117 (Betts). In Aim 3, we will define the ability of CD8+ T cells from eCLEAR participants who did or did not exhibit post-ART control to mediate viral control in vivo using a novel humanized mouse model. In addition, we will modulate CD8+ T cell functions within this system to uncover specific control mechanisms. The anticipated outcomes of our project are i) A comprehensive characterization of the immunologic and reservoir features associated with the partial efficacy of eCLEAR, ii) Validation of underlying mechanisms, to guide future iterations of clinical interventions at the time of ART initiation.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The goal of this proposal is to study and control rhythmic nephron induction for kidney replacement tissues. Kidney organoids derived from human induced pluripotent stem cells (iPSCs) re-create an astonishing cellular diversity comparable to the early fetal kidney. However, barriers remain to their implementation for regenerative medicine, chiefly the staggering volume of properly ‘plumbed’ tissue that would be necessary for functional replacement. This creates an urgent need to achieve scale-up of nephron generation. The kidney achieves scale-up during its development through an exponential increase in the number of nephron-forming ‘niches’ associated with the branching tips of the future urinary collecting duct tree. However, organoids generate nephrons in a single wave, failing to capture rhythmic, exponential nephron production from self-sustaining niches. Our long-term goal is to construct ‘higher-order’ synthetic kidney tissues using human stem cell, cell engineering, and assembly technologies that mimic the outcomes of morphogenesis. Our overall objective here is to gain temporal control over rhythmic nephron formation. Achieving this will mark a transformative advance toward creating replacement kidney tissue and in fundamental understanding of low nephron endowment, a risk factor for hypertension and chronic kidney disease. Our central hypothesis is that the periodic avalanche-like commitment of nephron progenitors to new nephrons is governed by a rhythmic ‘pace-maker’ across several signaling pathways. We plan to achieve our objective through two specific aims. Firstly, we will determine pace-making coordination across cell types in the mouse nephrogenic niche. We will expand from our preliminary spatial RNA sequencing data that discovered rhythmic alternating phases of nephron progenitor differentiation and renewal in each niche. This will define a spatiotemporally resolved map of cell-cell interactions contributing to nephrogenesis pace-making. Second, we will synthetically engineer pace-making and the nephrogenesis chain-reaction in human iPSC-derived organoids. We will program rhythmic nephrogenesis in iPSC-derived nephron progenitors using optogenetics technology and by leveraging intrinsic molecular clock dynamics. The proposed research is innovative because we co-opt our discovery of cyclical nephrogenesis signaling for novel engineering control strategies, while creating tissues that are compatible with patient-derived autologous cells for future transplantation. The proposed research will have significant positive impact in two areas: 1) Scale-up of human kidney tissue will create a step-change in renal replacement technology beyond dialysis, transplant, and “abiotic” filtration. 2) New discoveries in rhythmic nephron patterning will inform actionable approaches to improve persistence of nephrogenesis and increase nephron endowment in neonates.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Unhealthy diets significantly contribute to major preventable chronic diseases including type 2 diabetes, obesity, heart disease and stroke, which disproportionally impact racial/ethnic minority groups and those with lower income.1–3 Key health organizations agree that policies are needed to support healthier diets, including ultra- processed food and beverage taxes, nutrition warning labels, and marketing of healthier foods.4 Seven U.S. cities have sugar-sweetened beverage (SSB) taxes, the FDA is currently considering a national front-of-package food labeling system, and several U.S. cities have implemented or proposed healthy checkout regulations. It is now well-established that SSB taxes substantially reduce SSB sales and intake5 and research indicates that SSB nutrition warning labels can significantly reduce SSB purchases, though most of those data are from one- time online or lab exposures.6–8 Although taxes and warning labels targeting SSBs have been successful at shifting behavior, there are many other ultra-processed food products that contribute to unhealthy diets.9 What is less well-known is whether a suite of healthy food policies that are expanded to target a range of ultra- processed foods can shift dietary choices and intake in meaningful ways. To advance our understanding of policies needed to support nutrition security and health, our overall objective is to examine the degree to which a suite of healthy food policies in online food retailers can increase the purchase and intake of healthy foods and beverages while reducing the purchase and intake of unhealthy ultra-processed foods and beverages. To accomplish this objective, we propose using innovative online grocery store and restaurant platforms to randomize participants to either: 1) control (no taxes, warning labels, or healthy checkout regulations on any products); or 2) a suite of healthy food policies (ultra-processed food and beverage taxes, front-of-pack nutrition labeling, and healthy check out regulations that restrict the promotion of ultra-processed products on the checkout page). We will recruit 300 adults with lower income across Houston and San Antonio, TX, and Philadelphia, PA to shop once per week for eight weeks in both our online grocery store and restaurant. Week 1 will be a baseline (control) week without interventions, followed by five weeks of the interventions. In the last two study weeks, we will introduce unhealthy food marketing (e.g., banner ads) into the online platforms to mimic what we expect industry will do to counter public health policy efforts. Participants will be given money to spend in these online platforms and purchases will be delivered to them via a real food retail store and restaurant. Participants will complete surveys at baseline, 6-weeks, and 8-weeks and will complete two dietary recall interviews during the baseline week and during the fifth intervention week (4 recalls total). The specific aims are: to evaluate the effects of three healthy food policies on purchases across online grocery store and restaurant settings (Aim 1) and dietary intake (Aim 2). The goal of Aim 3 is to understand the degree to which unhealthy food marketing counters the effects of the healthy food policies.

NSF Awards · FY 2024 · 2024-08

This project aims to advance the mathematical analysis of non-linear partial differential equations used in a wide range of applications. The first part of this project involves the study of fluid dynamics problems with free boundaries, aimed at enhancing the understanding of water waves, tsunamis, and hurricanes. The second part of this project investigates the dynamics of gasses and plasmas under physical kinetic boundary conditions, which is expected to provide insight into important physical phenomena such as the solar wind, galactic nebulae, and the Van Allen radiation belt. The third part of this project explores the physical interactions between relativistic kinetic theory and gravitational models bringing potential to increase knowledge in astrophysics, such as in systems of galaxies, supernova explosions, models of the early universe, and the study of hot gases and plasmas. This project will support the education and training of postdoctoral researchers, graduate students, and undergraduate students through research mentoring and seminars. It aims to further the goal of developing a diverse and globally competitive STEM workforce and to improve STEM education at the collegiate level. This research will focus on improving the local-in-time well-posedness for large initial data and the global-in-time well-posedness near equilibrium for various fundamental non-linear partial differential equations. It involves developing new methods for analyzing several different physical models. One part of this work is to study fluid dynamics problems with free boundaries, such as the study of the Muskat bubble problem in 2D and 3D. Another part of this work examines problems related to the non-cutoff Boltzmann equation and the Landau equation from kinetic theory with the physical kinetic boundary conditions. The third part studies the relativistic Boltzmann equation and the Einstein-Boltzmann system. These developments are expected to benefit both mathematical and physical research in the future. 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 · 2024-08

Idiopathic hypersomnia (IH) is a poorly understood disorder characterized by excessive daytime sleepiness despite normal nighttime sleep, often accompanied by difficulty awakening, unrefreshing sleep, and cognitive impairment. Once thought to be present in <1% of the population, recent work indicates a far greater prevalence, with underdiagnosis exacerbated by a limited understanding of this disorder. IH patients commonly report poor management of symptoms and impaired quality of life, and treatments are not based on disease etiology. Twin studies show that daytime sleepiness is between 37-48% heritable and family history of excessive sleepiness, IH, or other hypersomnia disorders is present in ~1/3 of IH patients. Genome-wide association studies (GWAS) of hypersomnia traits have identified 24 genome- wide significant SNPs specifically associated with increased sleep propensity. Here, we propose a comprehensive approach to identify effector genes for hypersomnia-related traits, using “variant to gene mapping to in vivo validation” that will intersect GWAS data with ATAC-seq and high-resolution promoter- focused Capture C neuronal datasets derived from human induced pluripotent stem cells (iPSCs) to identify potential effector genes. We will rapidly validate identified candidates in an established Drosophila model of sleep and extend findings into zebrafish to test for behavioral relevance in a vertebrate system. Using approaches described in this proposal, we have already strongly implicated a gene, vertebrate CADM2, as associated with hypersomnia-like phenotypes across species with loss of function. In Aim 1, we will use a comprehensive combination of TAD-wise analysis of genome wide significant sleep propensity loci and “3D Genomics” approaches in key cell types to implicate additional candidate effector genes harbored in the sub-threshold P-value zones of the existing GWAS datasets. In Aim 2, we will use Drosophila to determine the mechanism through which loss of CADM2 (functional homolog in fly, beat-Ia) affects the function of key arousal circuits in the brain. In parallel, we will screen identified candidate genes for relevance to excessive sleepiness in vivo in flies using an RNAi-based approach. In Aim 3, we will rigorously examine sleep phenotypes in Cadm2b knockout zebrafish and prioritize other candidate genes for vertebrate using a rapid CRISPR approach pipeline. This project will identify novel genetic variants and the corresponding effector genes that contribute to hypersomnia-related traits, thereby shedding light on the biological pathways that influence the development of the traits. Study results will have fundamental implications for novel approaches to the diagnosis, prevention, and treatment of IH.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Pulmonary hypertension (PH) is a devastating lung disease characterized by pulmonary vascular remodeling and associated with high morbidity and mortality. Discovery of new therapeutic targets in PH has been limited by the heterogeneity in disease endotypes and incomplete mechanistic insight into the cellular processes that control and perpetuate vascular remodeling. Recently, the mechanistic target of rapamycin (mTOR) signaling pathway has been implicated in the development and progression of PH, including pulmonary arterial hypertension (PAH). Despite the growing importance of mTOR activation in pulmonary diseases, the effects of mTOR hyperactivation on pulmonary cells and the contribution of mTOR to the development and progression of vascular remodeling remains incompletely understood. In my preliminary data, I demonstrate that mesenchymal cells will act as a signaling hub after selective deletion of tuberous sclerosis complex 2 (Tsc2) and subsequent mTOR activation in progenitor mesenchymal cells (Tbx4LME-CreTsc2Mesenchymal KO). These cells orchestrate dysregulated cellular responses in arterial endothelial cells (ECs), leading to pulmonary vascular remodeling and spontaneous PH in Tsc2M-KO mice. This proposal will expand on this finding and examine how mTOR activation in specific mesenchymal populations alters EC behavior, function and trajectory and how aberrant mTOR signaling contributes to pulmonary vascular remodeling. To accomplish this task, I will utilize three novel mouse models with mTOR activation in distinct mesenchymal populations. To understand the disease relevance of the murine models, I will be performing concurrent experiments on primary lung tissues from lymphangioleiomyomatosis (LAM) and PAH. LAM is a monogenic disease with mTORC1 activation in lung mesenchyme cells. In additional to parenchymal changes, a subset of patients with LAM develop pulmonary vascular remodeling and PH. As a monogenic disease, LAM is an ideal model for the study of mTOR activation on EC biology and pulmonary vascular remodeling. This proposal leverage my expertise in pulmonary vascular biology, ex vivo model systems and in vivo experience with transgenic murine lines to generate significant discoveries regarding the fate regulation and function of endothelial cells in pulmonary vascular remodeling. My training, composed of coursework/workshops in bioinformatics and epigenetics, will take place under the mentorship of Dr. Vera Krymskaya, a leader in mTOR research. To ensure that I am meeting research and academic milestones, I have compiled a diverse but complementary Advisory Committee. Upon completion of this work, I will develop additional comprehensive skills in epigenetic analysis and advanced bioinformatics, ex vivo lung modeling, and in vivo murine models. Together, the research and career development plans proposed herein will facilitate a better understanding of the cellular crosstalk in the pulmonary microenvironment while preparing me for my future career as an independent investigator in the field of vascular biology.

NIH Research Projects · FY 2025 · 2024-08

Recent years have seen a surge of interest in the role of NAD in human disease and whether increasing NAD with supplements such as nicotinamide riboside (NR) or nicotinamide mononucleotide (NMN) has therapeutic value. However, our understanding of NAD and its metabolites in vivo is limited by technical barriers, a lack of consensus on analytical techniques, and incomplete knowledge of the catabolic routes taken when NAD breaks down under various stressed states. For many studies, blood and urine are the only samples available to investigative teams. Whole blood contains NAD(P)(H), but poses important stability considerations for handling and measurements. Serum and urine are relatively stable, but contain mostly catabolites of NAD, which themselves carry implications for NAD metabolism in the rest of the body. Here we propose to take advantage of a multidisciplinary team to develop and standardize methods for the analysis of NAD, as well as NADH, NADP, NADPH that are suitable for human clinical trials. These include extensive validation and testing of methods for determining the abundance of NAD(P)(H) in blood-derived samples and comparisons between liquid chromatography-mass spectrometry-based assays, biochemical assays, and nuclear magnetic resonance. We will also perform the first direct comparison of the two major magnetic resonance spectroscopy (MRS) methods that have been used to detect NAD in intact tissues of living subjects. In years 1 and 2, we will focus primarily on method development, establishing techniques for the optimal handling and storage of blood, as well as comparing the available methodology for NAD(P)(H) measurement. In years 3-5, we will conduct a human clinical trial, examining NAD(P)(H) levels in tissues and blood, along with catabolite levels in serum and urine before and after an intervention with nicotinamide riboside, thought to increase whole blood and tissue NAD levels. By performing all of these assays in the same individuals, we will also provide the first comprehensive assessment of the value of accessible bodily fluids for predicting tissue NAD levels and, more broadly, for detecting physiologically relevant alterations in human NAD metabolism. .

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY / ABSTRACT There has been considerable concern regarding the long-term effects of traumatic brain injury (TBI), largely driven by reports of neurodegenerative proteinopathies associated with participation in contact sports. In addition, increasing epidemiological data indicates that even a single moderate or severe TBI can increase the risk of dementia. While there has been a particular focus on the role of tauopathies following TBI, our group and others have demonstrated often complex and diverse neuropathologies with late survival post-injury in some individuals. However, the mechanisms driving these pathologies are poorly understood. In addition, to date there has been limited success in attempts to model both the neuropathological and clinical features of post-TBI neurodegeneration. As such, in order to permit mechanistic and translational studies of TBI-related neurodegeneration, there is a pressing need to develop preclinical models that recapitulate the complex pathologies and clinical phenotypes in the chronic phase post-injury. To address this critical knowledge gap, we propose an iterative design to develop novel gyrencephalic models of post-TBI ADRD. Three novel and distinct injury paradigms are proposed to represent repetitive mild exposures, as well as diffuse and focal TBI. We propose carefully considered biomechanical inputs relevant to real-world scenarios, with rigorous characterization of the nature and progression of neuropathological outcomes up to 1 year post-injury. The clinical relevance of pathologies will be determined via parallel and standardized comparisons with well-characterized human tissue samples from the CONNECT-TBI archive. Face validation will be determined through novel cognitive assessments, with additional validation attained via measurement of advanced neuroimaging and blood biomarker outcomes. Critically, reproducibility will be assessed at a second site. If successful, the development of a model with validated clinical relevance will provide a vital platform to study TBI-related neurodegeneration, including the assessment of novel diagnostics, and potential therapeutic interventions. Moreover, studying the evolution of pathology following the initiating injury may offer critical insights to the pathogenesis of TBI-related neurodegeneration.

NIH Research Projects · FY 2025 · 2024-08

A unifying pathology of age-related neurodegenerative diseases such as Parkinson’s disease (PD), Alzheimer’s disease (AD), Huntington’s disease (HD), and amyotrophic lateral sclerosis (ALS) is the accumulation of misfolded proteins and protein aggregates in the nervous system. Paradoxically, aggregation- prone proteins are expressed in both neurons and astrocytes, yet neurons appear more vulnerable to the accumulation of protein aggregates. These observations suggest cell-type-specific differences in how quality control pathways are managed in neurons and astrocytes to regulate the integrity of the proteome. In fact, we have found striking differences in how the autophagy-lysosomal pathway and the ubiquitin-proteasome system are managed between neurons and astrocytes. Despite this knowledge, a comprehensive understanding of the cell-type-specific pathways for regulating proteostasis in neurons versus astrocytes has not been achieved. Moreover, the relationship between neurons and astrocytes in the context of proteotoxic stress is largely unknown. It is well established that astrocytes play critical roles in neuronal homeostasis, function, and neuroprotection. By contrast, astrocytes are also emerging as key players in disease pathogenesis. However, we are only at the inception of understanding the molecular and functional properties of neuron-astrocyte interactions in health and disease. Thus, this proposal will define the regulation of lysosome-mediated pathways (autophagy and endolysosomal) within (Aim 1) and between (Aim 2) astrocytes and neurons in response to proteotoxic stress elicited by fibrillar forms of a-synuclein, a model for a-synucleinopathies including PD. To elucidate cell-type-specific responses, we developed a robust system to coculture primary neurons and astrocytes that recapitulates morphological, proteomic, and functional signatures of astrocytes in vivo. Based on our preliminary data, we hypothesize that autophagy and lysosomal pathways are adapted differently in astrocytes versus neurons, which may render neurons preferentially vulnerable to proteotoxic stress associated with a-synuclein aggregation. To test this hypothesis, we will (1) define the sequence of degradation of a-synuclein fibrils via lysosome- mediated pathways within astrocytes as compared with neurons, and (2) define the role of the autophagy receptor TAX1BP1 in the coupling of neuron-astrocyte stress responses to a-synuclein proteotoxins. We employ live-cell imaging combined with quantitative cell biology and biochemistry to investigate these processes at high spatiotemporal resolution in a compartment-specific manner. These studies will reveal new concepts in how quality control pathways are managed in the neuron-astrocyte unit in response to proteotoxic stress. This information will also inform cell-type-specific vulnerabilities in neurodegenerative disease and enable more specific strategies to mitigate neuronal dysfunction and death.

NSF Awards · FY 2024 · 2024-08

This award will fund research that studies the impacts of the development of improved predictive power of genetic testing on insurance markets. While scientific advances have led to increased predictive power of genetic data, many jurisdictions prohibit insurers from using genetic information to risks in underwriting, a situation that leads to consumers who know they are genetically predisposed to develop certain diseases decide to buy more insurance forcing insurers to raise prices for all. This research project combines large-scale genetic data, economic theory, and econometrics to study the impact of future genetic prediction technology on adverse selection in the market for critical illness insurance, and the more complex health, life, and long-term care insurance markets. In addition, the project will provide estimates of the future predictive power of genetic information for a large set of diseases, beyond what can be predicted by non-genetic risk factors. Results of this research will help decision makers to design better practices to ensure better functioning of insurance markets. The results will also provide input into practices to protect the most vulnerable in health insurance markets. This award funds a research project to study the effects of improved genetic testing on the functioning of health insurance market. The PIs first develop a method to measure the amount of adverse selection that would take place in a given insurance market if consumers had access to their current genetic information. The PIs the combine this estimates with estimates from heritability studies to estimate the amount adverse selection that would occur with future genetic prediction technology. These studies are based on a large-scale data on genetics, non-genetic risk factors, and insured events. Third, the researchers apply this method to existing insurance markets to measure adverse selection with both current and future genetic predictions. Finally the project augments the methodology with survival analysis specification and applies it to a large set of diseases and to project future predictive power of genetic information above that of observed covariates. The results of this research will provide inputs into the design of better practices to ensure efficient functioning of insurance markets. The results will also provide input into practices to protect the most vulnerable in health insurance markets. 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 · 2024-08

Project Summary The genetic landscape of patient samples has illustrated a vast array of potential diagnostic and clinical biomarkers through profiling of chromosomal rearrangements and examining subsequent alterations in RNA abundance. However, genome-wide analyses have also discovered that there are discrepancies between transcript expression levels and corresponding protein abundance, highlighting the limited predictive power of bulk RNA-based prognostic strategies to assist in therapy. Moreover, there are an increasing number of studies depicting thousands of missed and unannotated peptides. These discoveries underscore the importance of understanding RNA regulation in the context of disease to showcase the complete landscape of genetic expression and target the phenotype of malignant cells. Our findings have noted that select mRNAs are translationally increased, independent of their transcriptional regulation, in response to disease progression, and can be studied to create novel therapeutic avenues unique per disease type. Mapping post-transcriptional regulation and the un-annotated proteome, including RNA localization and modifications to assist in translational regulation, however, remains poorly understood. As cells rely on the dynamic response of protein synthesis to quickly respond to environmental cues, this leaves an unexplored area for discovery that is required for cell survival and is promising for innovative therapies. We have identified essential signaling advantages activated to rewire RNA translation during initiation and for the maintenance of aggressive disease states. During these processes, ribosome initiation is altered and increased at non-canonical start sites, which can alter protein variants and produce micropeptides. Our research focuses on mechanistically determining how growth and adaptive signaling cascades interact by: 1) mapping the post-transcriptional aspects of gene expression missing from our current annotated genomes, focusing on alternative ribosome initiation, and 2) investigating the RNA-modification and binding proteins influencing selection and spatial interactions of RNA. This regulatory control rapidly reprograms the cellular phenotype at the level of RNA translation, increasing genetic diversity, and we will be able to characterize the global utilization of nonconventional initiation to study cell state changes modeled during cancer progression. Our multidisciplinary approach is designed to connect unbiased ribosome sequencing with proteomic profiles to identify the complete oncogenic proteome and the features that impose RNA selection. We will identify the trans- factors, cis- domains, and RNA modifications utilized by adaptive signaling to commandeer the translational response between cell states. Our studies will provide unprecedented insight into RNA regulation through focusing on localization and modifications alongside ribosome initiation, and how their interplay alters gene expression. This will lay the foundation for mapping genetic diversity through noncanonical translation.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Improving outcomes for adult depression in primary care is a leading health care priority and antidepressants are a cornerstone of evidence-based treatment. However, patient non-adherence and discontinuation are common. The proposed study examines use of modest financial incentives in conjunction with tailored text message reminders to determine the most effective and cost-effective support for establishing consistent antidepressant medication taking routines to improve clinical outcomes of adult primary care patients with depression. We propose to conduct a 3-arm RCT (N=525, n=175 per arm) to compare with usual care the short-term and extended effectiveness of two adherence support strategies for primary care patients with depression who have been newly prescribed antidepressant medications. We will examine whether personalized daily text messages with and without financial incentives improves antidepressant adherence and depression symptoms. Adherence will be measured with a wireless pill bottle at 6 and 12 weeks and by assessment of electronic health prescription records at 24 and 52 weeks. Depression symptoms will be collected via telephone by a trained assessor at 6 and 12 weeks. Our Specific Aims are to: 1. Determine the relative effectiveness of 1) 12 weeks of personalized daily text reminders without financial incentives (reminders alone), 2) 12 weeks of the text reminders paired with 6 weeks of financial incentives (reminders and incentives), compared with 3) usual care, and with each other (non-inferiority). The primary outcome will be symptom response on the PHQ-9 depression rating scale at 12 weeks. 2. Explore the effectiveness of each study intervention arm compared to usual care and to each other on antidepressant adherence at 6, 12, 24, and 52 weeks, and whether these effects are moderated by baseline financial security and adherence intentions. We also will assess whether the intervention effects in Aim 1 are mediated by antidepressant adherence. 3. Use qualitative inquiry of antidepressant adherent and non-adherent study patients to explore opportunities to maximize the effectiveness of the financial incentives and reminders to increase antidepressant medication adherence. Widespread problems with antidepressant adherence, especially during the early stage of treatment, undermine the primary care treatment of depression. This study will test whether personalized daily text messages grounded in behavioral economics principles alone or combined with financial incentives based on contingency management principles improves depression outcomes. These two interventions are designed to be readily adapted into primary care workflows through an automated patient-facing system to improve clinical outcomes of adult primary care patients with depression.

NSF Awards · FY 2024 · 2024-08

New York University, Northwestern University, and the University of Pennsylvania form the Critical STEM Faculty Alliance (C-STEM). Leveraging their combined strengths, they aim to develop an infrastructural technology system that provides more opportunities and lowers systemic risks for historically underrepresented groups. C-STEM will examine college and university functions, to understand how to train effective technology researchers and teachers from these groups. NSF emphasizes creating opportunities everywhere. Accordingly, C-STEM seeks to help new researchers from underrepresented backgrounds build strong professional networks, establish stable pathways that advance careers, and collaborate with other experts in academia, industry, and government. Also, C-STEM aims to help researchers build new projects and design innovative educational tools to improve people's lives. Its goal is to ensure that technology serves the public interest, especially those who have been most negatively affected by technology. C-STEM aims to design and implement institutional self-assessments at the three C-STEM Alliance institutions. The alliance will prioritize collecting and analyzing data to identify inequities affecting underrepresented minority (URM) doctoral students, postdoctoral scholars and early career faculty in STEM fields. To assess the need for the C-STEM Alliance, the project will collect data on the demographic representation (race, ethnicity, national origin, sex, gender, first-generation status) of doctoral students and faculty in STEM and related fields. The project will also conduct curriculum surveys to understand demographic and socio-technical content representation in STEM courses, and review research production by minority and non-minority STEM students and faculty. Surveys will also evaluate existing mentorship and support structures, and collect data on minority STEM doctoral student outcomes, such as degree completion and post-degree hiring. Additionally, the alliance will gather qualitative data from minority STEM students and faculty about their experiences. This data will help identify institutional challenges and justify the alliance's activities, demonstrating how they address specific needs. To assess institutional readiness, the project will collect data that include reviews of diversity commitments by university leaders and progress towards diversity, equity, and inclusion goals. This will demonstrate C-STEM institutions’ commitment to increasing the representation, resilience, and success of minority doctoral students and faculty in STEM. The alliance intends for this work to help research communities better understand the incentives and affordances institutional leaders’ encounter in their efforts to create, continue, or expand key structures, such as postdoctoral programs and frameworks for transitioning postdoctoral scholars to tenure-track positions. 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 · 2024-08

ABSTRACT Impaired ability to respond to the earth’s gravitational field and loss of balance results in high risk of falls with adverse consequences for one’s health, quality of life, and health care costs to society. Falls are a particular risk for individuals with Parkinson’s disease as well as elderly humans in general. Despite the importance of gravity sensing to daily activities, the molecular, cellular, and circuit gravity sensing mechanisms are incompletely understood. To understand gravity sensing mechanisms at the molecular and neural circuit level, we use the simple microscopic nematode Caenorhabditis elegans. C. elegans offers many experimental advantages, including a small and simple nervous system, accessibility to rapid genetic manipulation and to optogenetic tools, and ease of cultivation. We have demonstrated that C. elegans senses and responds to gravity, and have used mutant analysis and pharmacology to show that one or more types of ciliated sensory neurons, as well as dopamine, play a role in gravitytaxis. Our preliminary data leads to the hypothesis, which we will test, that one or more of the three ciliated dopaminergic neurons types—CEP, ADE, and/or PDE— mediates the response to gravity. We will use genetically-encoded synaptic transmission toxins as well as genetically-encoded chemogenetic experiments to determine which of these 3 neuron types is required for the response to gravity. We will identify additional neurotransmitters required for gravitaxis by testing animals mutant for each of 4 neurotransmitter systems. We will test each of 58 C. elegans genes predicted to encode mechanosensitive ion channels for a role in gravitaxis. Finally, we will make use of the million mutation project to perform a pilot unbiased genetic screen for gravitaxis-defective mutants. In addition, we will develop a high throughput sorter to isolate gravitaxis – deficient mutants for a future high throughput genetic screens for additional gravitaxis defective mutants. At the completion of these studies, we will have identified one or more dopaminergic neuron type as well as other neurotransmitters required for gravitaxis and we will have developed the tools needed for further studies consistent with the objectives of this R21 exploratory/development funding mechanism. Our work will contribute to a comprehensive molecular understanding of animals’ response to gravity and set the stage for the development of novel therapeutics to reduce falls.

NIH Research Projects · FY 2025 · 2024-08

Project Description The advancement of regenerative medicine has provided many new potential treatments for craniofacial bone defects, in which mesenchymal stem/progenitor cells (MSCs) play a critical role in maintaining constant remodeling of tissue architecture. The neural crest derived orofacial MSCs (OMSCs), such as stem cells from apical papilla (SCAP) are attractive postnatal stem cells for hard tissue regeneration, based on their superior osteogenic properties compared to their bone marrow counterparts. However, clinical translation remains challenging due to limited knowledge about the mechanisms of action. Ferumoxytol, an FDA-approved iron oxide nanoparticle formulation, exhibits several biomedical properties including anticancer and immunomodulation, based on its inherent physicochemical properties that activates cell proliferation, migration, and differentiation. Given these properties, ferumoxytol could be applied in MSC-based tissue regeneration, an unexplored avenue. Specifically, our goal is to identify whether ferumoxytol can activate orofacial MSCs and promote their multipotent differentiation capabilities and immunomodulation for endogenous tissue regeneration. Using RNA sequencing (RNA-seq) analysis and in vitro MSC characterization, we found intriguing data demonstrating that: 1) ferumoxytol significantly promotes stemness of SCAP through elevation of MSC markers and osteogenic progenitor markers, 2) proliferation and osteogenic capabilities are highly activated in SCAP after ferumoxytol treatment, 3) ferumoxytol largely increased immunomodulation of SCAP via PGE2/IDO cascades, and 4) YAP/TAZ are required mediators in ferumoxytol-mediated metabolic reconfiguration of SCAP for tissue regeneration. Based on these findings, we hypothesize that ferumoxytol activates YAP/TAZ signaling and promotes stemness of orofacial MSCs, which provides a favorable physiochemical microenvironment for enhancement of MSC viability and osteogenesis for autotherapies. During this proposal, I will explore the role of ferumoxytol in activation of somatic OMSCs to address how YAP/TAZ-mediated metabolic switch regulates OMSC homeostasis (Aim 1). Since immune components can impair somatic stem cell function and ferumoxytol treatment significantly elevated cell proliferation and osteogenesis through activation of YAP/TAZ cascades, we will determine whether activation of YAP/TAZ by ferumoxytol treatment may elevate endogenous craniofacial tissue regeneration in a ligature- induced periodontitis mouse model. A tetracycline-inducible postnatal neural-crest-specific YAP/TAZ knockout mouse model will be generated to examine in vivo stem cell behavior and endogenous tissue regeneration ability (Aim 2). Upon successful completion of the Specific Aims, this translational study will extend our knowledge in activating somatic stem cell abilities through repurposing an FDA approved nanoformulation for a new biomedical application.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Pre-eclampsia is an obstetric condition that affects 5–8% of all pregnancies and is a leading cause of maternal mortality worldwide. Pre-eclampsia is characterized by the onset of maternal hypertension after 20 weeks of gestation, stemming from insufficient remodeling of the uterine spiral arteries that supply blood flow to the placenta and fetus. Clinical treatments for pre-eclampsia, such as antihypertensive drugs to manage blood pressure and anticonvulsants to prevent seizures, address the associated symptoms, yet no drug or therapeutic has been developed to slow the progression of pre-eclampsia. The only curative treatment option for pre- eclampsia is the early delivery of the placenta and fetus, which often resolves maternal hypertension within a few days, but can cause fetal morbidity and mortality, especially in cases where fetal growth restriction occurs concurrently with pre-eclampsia. Therefore, a significant unmet need exists for novel therapeutic strategies that can facilitate vasodilation in the placenta during pre-eclampsia to resolve maternal hypertension and improve fetal health. With no drug available to slow disease progression, engineering ionizable lipid nanoparticles (LNPs) for extrahepatic mRNA delivery to the placenta is an attractive therapeutic platform to treat pre-eclampsia. The goal of the proposed work is to develop a targeted, placenta-tropic LNP platform for the delivery of mRNA cargo to the placenta to treat pre-eclampsia during pregnancy. First, high-throughput in vivo screening will be utilized to evaluate a large library of LNPs with novel ionizable lipid structures and excipient compositions for extrahepatic tropism to the placenta. Lead candidates from this high-throughput screen will be functionalized with active targeting motifs, namely placenta-specific antibodies, for selective delivery to trophoblasts and endothelial cells in vivo. Following enrichment analysis using next generation sequencing, LNPs demonstrating placental tropism will be functionalized with placental-specific antibodies to promote active targeting to key cell types in the placenta — trophoblasts and endothelial cells. In vitro, ex vivo, and in vivo expression of therapeutic mRNA cargos (VEGF, PlGF, and eNOS) encapsulated in LNPs will then be evaluated in primary human and mouse placentas. The lead mRNA LNP candidates will then be utilized to rescue maternal hypertension, fetal health, and immunophenotype in an induced model of severe, early onset pre-eclampsia in mice. The work proposed here is the first to evaluate the therapeutic efficacy of a pro-angiogenic mRNA LNPs for treating pre-eclampsia, a placental disorder during pregnancy for which currently no curative treatment options exist. Due to the modular nature of LNPs and the ability to readily swap mRNA cargoes, the platform developed through the completion of these studies can be used for treating not only pre-eclampsia, but also a wide range of placental disorders occurring during pregnancy.