University Of Pennsylvania

NIH Research Projects · FY 2025 · 2022-12

Project Summary/Abstract: Accurate repair of DNA damage is critical for genetic stability, and for preventing aging-related degeneration and cancer. We are working to identify key factors that regulate accurate repair of DNA double-strand breaks (DSBs) through the error-free homologous recombination (HR) pathway. DSBs can arise from many sources including endogenous replication fork damage, exogenous environmental toxicants, or oxidative stresses induced by endogenous sources and during pro-inflammatory responses to toxicant injury. We found that the RAD51 paralogs are critical for promoting HR and hence for suppressing error-prone repair mechanisms. Over 300 studies link mutations in human RAD51 paralogs with cancer, and women with breast or ovarian cancer are now screened for RAD51 paralog mutations. However, it remains largely unknown which RAD51 paralog mutations are pathogenic and how these mutations sensitize individuals to environmentally induced-DNA damage due to our lack of functional analysis of either the wild-type or mutated proteins. We do not know how these proteins are recruited, their functional components, or the disruptions caused by mutations or polymorphisms in the RAD51 paralogs. This knowledge gap results from low abundance of endogenous RAD51 paralog proteins, insolubility of the recombinant proteins, as well as embryonic lethality in knock-out mice. We are therefore using genetic, biochemical, and cell biological approaches to characterize RAD51 paralog function upon exposure to DSB inducing agents. We will use ionizing radiation (IR) and bleomycin as model agents for environmentally relevant DSB-inducing agents. Using complementary approaches in combination with high-throughput genetic screening, we are now uniquely poised to address how RAD51 paralog mutations predispose individuals to human cancer and thus, to identify opportunities for determining who is at risk for cancer development upon exposure to environmental carcinogens. Our ultimate goal is to enable development of precision medicine strategies for individual patients whose tumors harbor a RAD51 paralog mutation profile.

NIH Research Projects · FY 2025 · 2022-12

PROJECT SUMMARY Alcohol use disorder (AUD) is one of the most prevalent psychiatric disorders in the United States. Chronic alcohol consumption in individuals with AUD is associated with lowered glucose metabolism and increased acetate metabolism in the brain. Following abrupt cessation of alcohol consumption, sudden decreases in acetate may cause the brain to become “energy-deprived”, which could induce heightened craving and symptoms of withdrawal. The main goal of this proposal is to test the hypothesis that energy deprivation during withdrawal contributes to alcohol craving by influencing reward-encoding neural circuitry. Analyses for Specific Aim 1 will focus on defining the impact of altered brain metabolism in the dorsal anterior cingulate cortex (dACC) on its connectivity with the ventral striatum (VS) and on specifying the role of connectivity between the dACC and VS in facilitating whole-brain transitions to alcohol cue-induced neural activity states. Analyses for Specific Aim 2 will seek to define the relationship between metabolic function in the dACC, its role in driving whole-brain dynamics, and alcohol craving. Completion of this proposal will provide a framework for predicting the impact of metabolic interventions on brain state and craving, which will aid the development of therapeutic approaches for AUD.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY Network gene analysis suggests that high confidence autism spectrum disorder (hcASD) genes operate through convergent mechanisms that predominantly support synaptic function, which, in turn, can determine anatomical and functional brain connectivity. Several variants in ANK2 have been identified in ASD patients and this gene has been consistently considered a top hcASD risk. ANK2 encodes ankyrin-B (AnkB), which roles in axonal biology may be an underlying factor in ASD pathophysiology. However, our recent findings indicate that ankyrin- B also plays important roles at the cortical excitatory postsynapse. Moreover, both predominant AnkB isoforms in neurons (220 kDa and 440 kDa AnkB) are abundantly expressed in dendrites, where they likely perform isoform-specific roles and interact with unique partners, including proteins encoded by other hsASD genes. Thus, it is important to define the isoform-specific roles and partners of AnkB at the postsynapse. To accomplish these goal we will 1) use in vivo proximity-dependent biotin identification (iBioID) combined with in tandem mass spectrometry (MS) to capture and identify AnkB interacting partners in the postsynapse by leveraging our novel biotin ligase BirA-tagged conditional AnkB (AnkB-BioID) knock-in mouse and isoform-specific AnkB knockout mice; 2) combine live and super-resolution microscopies and cellular assays to validate compelling AnkB interactors; and 3) define the significance of novel AnkB PSD complexes for synaptic development and function.

NIH Research Projects · FY 2026 · 2022-11

Project Summary Kidney transplantation is the preferred treatment of patients with end-stage renal disease compared to dialysis in terms of patient survival, quality of life and cost. One of the most common causes of premature graft loss after kidney transplantation is alloimmune-mediated injury. HLA alloantigens represent a significant barrier to long-term allograft outcome. Kidney allograft failure is often caused by recipient immune recognition of foreign human leukocyte antigen (HLA) proteins of the donor organ. The HLA gene complex comprises multiple loci and has a high degree of genetic polymorphism. HLA amino acid (AA) polymorphisms strongly impact key structural and functional features of HLA molecules, including allorecognition by T cells and alloantibody. Previous studies have shown that mismatches (MMs) at HLA antigens are associated with worse outcomes, with the highest risk of graft failure (GF) particularly associated with HLA-DRB1 MMs. However, the relative impact of AA variation is unknown because organ allocation systems have not collected comprehensive molecular HLA typing data. Single-center studies using high resolution HLA typing to evaluate MMs at surface- exposed amino acids (termed “eplets”) have shown that the number of MMs correlates with the presence of de novo donor specific antibodies (dnDSA) and GF. However, these studies involved relatively few subjects and were not representative of the ethnically diverse transplant population. In addition, these studies assumed monotonic risk increase, a gap that we plan to address in this grant application. We will use HLA imputation methods that we have developed to unlock the capability to utilize the SRTR database for association analysis of AA MM categories with graft failure. In addition, we will validate and further investigate AA MM associations using a large multi-center cohort of kidney transplants wherein high resolution HLA class I and class II typing is readily available. We will also evaluate associations of HLA AA MM assortments with risk of dnDSA development. This project aims to answer several unresolved questions about HLA AA MMs and outcomes: (1) Which HLA loci are most important to match? (2) On top of antigen-level mismatches, can amino acid level mismatches further stratify outcomes? (3) Are some assortments of AA or AA motifs more important to match for than others? Our team has developed a machine learning feature engineering to discover and optimize AA MM groupings (bins) associated with GF and dnDSA. Improved patient risk stratification may help identify which transplant recipients would benefit from less aggressive immunosuppressive regimens and by reducing the number of repeat transplants due to graft failure with a poorly matched donor. Our approach generalizes broadly to other organ transplantation and possibly to hematopoietic cell transplantation.

NIH Research Projects · FY 2025 · 2022-09

Summary This application seeks support to develop the next generation of the Advanced Normalization Tools (ANTs) ecosystem, an open-source library spanning multiple software packages and programming languages that houses top-performing algorithms used worldwide by a large community of researchers. The core software library, ANTs, is built upon and contributes to the well-known NIH-funded Insight Toolkit (ITK), which has historically been a significant resource of open-source algorithmic and software innovation. Over the course of its decade-long development, ANTs has enabled hundreds of academic and industrial scientists to meet modern quantitative imaging needs with particular focus on issues in biomedical imaging-a broad range of applications and published research literature sample the study of organisms from small animals to humans as well as target organ systems such as respiratory, cardiovascular, and nervous. Today, ANTs is a freely available, cross-platform toolkit for multiple modality image processing which continues to set the standard in the field. However, despite its success in meeting the real-world needs of a broad community of investigators, the rapidly growing use of imaging, and the associated need to solve ever more complex image registration problems, require a dynamic update of ANTs in order to meet current and emerging needs of the biomedical research community. This major update, representing the first ever application for dedicated funding support of ANTs development, will comprise of the four specific aims of this project: 1) to significantly broaden the range of multiple modality registration tasks able to benefit from AN Ts-based solutions; 2) to modernize and expand the tool's support of very large datasets and scalable computing platforms; 3) to provide reference registration protocols and validate their reliability in investigator-driven research studies; and 4) to enhance support and outreach to the user community. These developments will ensure the long-term viability of ANTs in the face of significant advancements in computing and imaging technologies, as well as enable an even broader research audience to leverage ANTs in their scientific output.

NIH Research Projects · FY 2025 · 2022-09

Project Abstract Apathy is the most common and disabling of the behavioral symptoms shared across many Alzheimer's Disease and Related Disorders (ADRDs), including behavioral variant frontotemporal degeneration (bvFTD). Apathy manifests as a decrease in goal-directed behavior (GDB), with deficits such as poor planning, poor motivation and inability to initiate even the simplest self-care activities, contribute to disability and greatly reduced quality of life. Thus, apathy is a poor prognostic indicator, having a profound impact on clinical decline in everyday patient functional activities. Recent work shows that apathy is associated with a disruption in impairments in GDB--initiation, planning and motivation--suggesting that apathy is a heterogenous syndrome with distinct underlying mechanisms. Furthermore, these functionally dissociable GDB processes map onto distinct and distributed brain regions. While previous imaging efforts have predominantly focused on the identification of structural MRI correlates of single brain regions involved in apathy, the overall goal of this proposal is to investigate large-scale functional networks underlying impaired GDB in bvFTD--where apathy is highly prevalent. Building on our previous work, the framework proposed here will capture the complex associations of impaired GDB encompassing the various domains of apathy and will examine the ways the breakdown of large-scale intrinsic networks can lead to the clinical syndrome of apathy. In Aim 1, we will study how distinct impairments in GDB contribute to rate of longitudinal clinical decline in everyday functional activities. In Aim 2, we will use resting-state fMRI to identify relationships between impaired GDB and breakdown of large-scale neurocognitive networks. In Aim 3, we will examine how change in configuration of functional network connectivity over time contributes to decline in components of GDB and we will assess how degrading networks underlying GDB mediate rate of clinical decline in everyday functional activities. This proposal addresses a critically unmet need to elucidate the role of degenerative disease in compromising the network mechanisms that support goal-directed behavior in ADRD. Given the limited effectiveness of pharmacological treatment for apathy in dementia, this translational work is necessary to guide future clinical trials for this debilitating syndrome.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT A rich network of sensory afferents in the ocular surface (OS), supplied by the ophthalmic branch of the trigeminal nerve, performs a multitude of physiological functions, including sensation, regulation of various reflexes, and secretion of trophic and growth factors. Of the OS structures, the cornea is the most highly innervated tissue in the body. As such, dysfunction of the corneal/OS nerves has been shown to underlie a wide range of diseases, including trauma, infections, metabolic imbalances, and therapeutic interventions such as refractive surgeries. Corneal diseases continue to be a major health problem in the US, and corneal nerve dysfunction contributes to many of these disorders, such as neurotrophic keratitis, corneal neuropathic pain, and dry eye disease (DED). DED alone has been shown to impact up to 75% of a given population, causing significant reduction in quality of life and increased risk for visually debilitating opacities. Therefore, a comprehensive characterization of corneal sensory nerves is critical for understanding the pathophysiology leading to OS diseases. To meet this challenge and in response to the FOA RFA-EY-21-004 on OS innervation, we have assembled an interdisciplinary team with a strong collaborative track record and complementary expertise, including eye disease models and wound healing (Lee); pain and the mammalian somatosensory system (Luo); intravital corneal nerve fiber imaging and regeneration (Rompolas); single-cell transcriptomics/epigenomics and related computational analyses (Wu); and neurophysiology and data analysis/machine learning (Ding). We propose to combine advanced imaging approaches, novel single-cell multi-omics, and cutting-edge mouse genetic models to perform three levels of analysis: 1) Morphology - using single neuron genetic labeling, we will resolve the fine morphology, spatial organization, and connectivity of corneal sensory neurons by visualizing axonal arborization in the cornea and brainstem/spinal cord. We will also examine interactions of individual sensory afferents and corneal epithelial cells. 2) Molecular - we will perform integrated single-cell transcriptomics and epigenomics to interrogate the subtype-specific marker gene expression and epigenetic landscape of the corneal sensory neurons. 3) Functional - we will perform real-time physiologic analyses of corneal sensory afferents using in vivo two-photon calcium imaging. To further elucidate the role of sensory nerves in OS pathophysiology, we will determine how their morphology, molecular profile, and responsiveness are altered in a surgically induced DED model. Moreover, we will implement a novel high-speed imaging and machine learning platform to quantify evoked responses to corneal sensory stimuli and spontaneous behavior in the DED model. Taken together, our study will generate a comprehensive data set poised for integration that will fill in critical knowledge gaps in the field, create a robust foundation for future studies, and inform the development of more precise therapeutic strategies to treat OS diseases.

NIH Research Projects · FY 2026 · 2022-09

Primary progressive aphasia (PPA), a debilitating condition of language loss associated with Alzheimer's Disease and Alzheimer's Disease Related Dementias (AD/ADRD) affecting many patients with frontotemporal dementia (FTD) and Alzheimer’s disease (AD), lacks effective treatments. One of the most common and burdensome impairments associated with this manifestation of AD/ADRD is anomia, the inability to access the names of objects, experienced by patients as word-finding difficulty. Transcranial direct current stimulation (tDCS), a form of noninvasive neuromodulation, shows promise as an intervention for anomia in persons with PPA. However, efforts to introduce this novel treatment approach into clinical practice are hampered by the modest size and scope of prior tDCS studies in this patient population, which prevents establishment of clear, robust clinical evidence to support the widespread use of tDCS in clinical contexts. To address this gap, this proposal aims to conduct the world’s first well-powered, multi-site Phase 2 clinical trial of tDCS therapy in PPA. In order to accomplish this goal, the current project solidifies a collaboration between research teams at Johns Hopkins, University of Pennsylvania, and Baycrest Health Sciences, which have collectively produced 100% of the published studies on tDCS in PPA in North America and 80% of the studies internationally. In a total cohort of 120 patients with PPA, we will employ a randomized, double-blind, within-subject, cross-over design similar to trials that have been previously employed at each of the contributing sites. Following baseline behavioral testing focused on language abilities and MRI imaging, subjects with PPA will receive 10 daily sessions over two weeks of either tDCS paired with NAming and SPelling treatment (NASP; a behavioral language therapy targeting both oral and written naming production) or sham (placebo) tDCS paired with NASP. Participants will undergo behavioral testing and imaging again at the end of the intervention, receive behavioral testing a month later, and then repeat testing and brain imaging 3 months after the end of the intervention. Participants will then switch study arms and receive the intervention, testing, and imaging at the same intervals. We will also conduct baseline behavioral testing and neuroimaging (with no treatment) on 60 neurologically healthy older adults for comparison to PPA subjects with respect to language performance and brain imaging. Subsequent analyses will determine whether tDCS over the left frontal language areas paired with naming treatment results in persistent improvement in oral and written naming, and will also identify the clinical, neural, biological, cognitive, and demographic characteristics that predict tDCS effects on naming performance. Owing to our unique opportunity to study a largely bilingual population in Canada, we will focus specific attention on bilingualism as a demographic feature that may influence outcomes. Overall, this impactful trial will provide the most reliable answer to date as to whether tDCS is effective for treating naming deficits in PPA and will help the field to identify patients best suited for this promising intervention in AD/ADRD.

NIH Research Projects · FY 2025 · 2022-09

People who use drugs (PWUD) with acute medical problems have high rates of subsequent mortality and morbidity related to substance use. While health promotion strategies to reduce overdose and injection-related complications have historically functioned outside of health care settings, integrating health promotion strategies into acute care clinical settings could improve these outcomes. There is a critical gap in how best to implement health promotion strategies for PWUD in the health system that will lead to effective behavioral change. Our study proposes to develop an acceptable, feasible, and effective peer-led bundle of health promotion strategies. Our overall objectives are to tailor the THRIVE (Teaching Health promotion and Resilience in Varied Environments) intervention and determine its efficacy in changing behaviors and reducing health risks among PWUD. The THRIVE intervention was conceptualized by our team along with people with lived experience and informed by the COM-B model for Behavior Change and the Theoretical Domains Framework. THRIVE includes a face-to-face session boosted by weekly text messages and electronic content. Content is delivered over 12 weeks. Our research team has the extensive expertise needed to successfully complete the proposed aims. We will use a human-centered design approach to tailor the THRIVE model and address the patient, provider, and systems-level barriers to implementation in hospital and emergency-department settings. This includes a “Design Sprint” in which PWUD will map the problem (guided by qualitative interviews with patients and healthcare providers), sketch implementation elements, choose the key aspects to develop, and build model components. We will then examine the efficacy of the THRIVE intervention in reducing the cumulative incidence of self-reported non-fatal overdose or skin and soft tissue infection between baseline and 6-months among PWUD in a hybrid type 1 randomized controlled trial. We will conduct a randomized controlled trial recruiting patients (n=390) admitted to the hospital or Emergency Department with opioid use disorder in one of three hospitals. We will also analyze implementation barriers and facilitators of the THRIVE model to identify any needed implementation supports for widescale implementation. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions to the national opioid public health crisis. The NIH HEAL Initiative bolsters research across NIH to improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2024 · 2022-09

Project Summary The goal of this proposal is to dissect the distinct molecular roles of protein-RNA interactions between RNA and the Polycomb Repressive Complex 1 (PRC1) during neural differentiation. PRC1 is a conserved chromatin modifier that represses transcription via chemical and structural alterations of chromatin architecture. PRC1 mutations cause neurodevelopmental disorders characterized by microcephaly, intellectual disabilities, and dysmorphic body features, making it crucial to gain understanding about PRC1 function. Despite its ubiquitous expression, PRC1 can target distinct sets of genes for silencing in different cell lineages, resulting in cell-type specific expression. Though there are decades of research on Polycomb protein function, a key unanswered question is how PRC1 selects different target genes in different cell types. Mounting evidence in the field indicates that major epigenetic events are regulated by interactions between RNA and chromatin-modifying proteins, including subunits of PRC1. However, PRC1-RNA interactions remain underexplored. My preliminary data indicates that at least two PRC1 subunits, SCMH1 and RING1B bind to RNA in vivo. I will investigate the regulatory role of these protein-RNA interactions in the context of directed differentiation of mouse embryonic stem cells (ESCs) into neural progenitor cells (NPCs) with the following specific aims. In Aim 1, I will identify RNAs directly bound to SCMH1 and RING1B in pluripotent ESCs using state-of-the art photocrosslinking and sequencing approaches. I will also map protein residues required for these interactions using a new technology that couples genetically encoded peptide barcodes to a mutational library for each subunit followed by a mass spectrometry readout. In Aim 2, I will investigate the functional roles of these protein-RNA interactions during neural differentiation by using our existing RNA-binding PRC1 mutants and a degrade-and-rescue approach, leveraging my lab’s expertise in acute protein degradation. The experiments in this proposal will provide the first in-depth characterization of PRC1-RNA interactions in regulating cell fate transitions during differentiation. To achieve these aims, I have developed with my sponsor and co-sponsor a rigorous and comprehensive training program with four primary goals: 1) become an expert in mass spectrometry for protein-RNA biochemistry, 2) gain experience in cutting-edge functional genomics methods, 3) increase proficiency in bioinformatics and data analysis, 4) sharpen my research communication skills. I am confident that my choice of sponsor and co-sponsor combined with my diverse training background and the collaborative nature of my training environment will enable me to achieve my goals and the proposed research plan simultaneously.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract Understanding how niches regulate stem cells is critical to human health because aberrant regulation by the niche can cause tumor formation or tissue atrophy. Well-studied niches form in predictable structures following reproducible morphogenetic changes, suggesting niche structure is regulated and functionally important. This has not been directly studied. Additionally, the mechanisms that determine the initial positioning of niches during development remain understudied. To address these deficits, we study the Drosophila posterior signaling center (PSC)—the niche of the larval lymph gland. The PSC maintains hematopoietic progenitors, and it induces differentiation of a special immune cell upon immune challenge. The PSC forms during embryogenesis: its cells migrate dorsally, where they ultimately reside, coalesced at the lymph gland posterior. Preliminary data indicates that mutants without visceral mesoderm (vm) have dispersed PSCs and fewer PSC cells. As such, the overall hypothesis is that the vm guides PSC positioning to the lymph gland posterior. We further hypothesize that a coalesced PSC is required for its optimal function as a niche. Aim 1 will identify the role of vm and its mechanism of action in instructing PSC formation via live-imaging of vm mutants to identify the timing and type of the PSC defect, and then testing vm candidate cues with vm-specific RNAi knockdown of the cue followed by analysis of PSC positioning. The PSC signal transducer will be identified by PSC-specific RNAi knockdown of candidate transducers. Ectopic expression of the vm cue in a nearby tissue will reveal if the cue is a true guidance cue sufficient to guide PSC positioning, or whether it confers competency to respond to other positional cues. Aim 2 will investigate how PSC coalescence contributes to niche function by causing PSC dispersion and then assessing PSC functions. Ability of the dispersed PSC to maintain progenitors will be tested by quantitating PSC- dependent progenitors, and ability to generate an immune response by challenging larvae with parasitoid wasp infection, and then quantitating a specialized immune cell type. The PSC can sense the organism’s nutrient and immune environment, but it is unknown if it senses feedback from the cells it regulates. This will be tested with lineage-specific ablation of progenitors or mature hemocytes in the lymph gland with Gal4/UAS-driven apoptosis. Then the level of PSC maintenance and differentiation signals will be measured for comparison to controls. Accomplishing these aims will reveal principles of niche formation, how niche structure impacts its function, and whether a niche receives feedback from the tissue it supports. Research training will take place at the University of Pennsylvania under the advisement of the Sponsor, the PI’s thesis committee, and Penn faculty as needed. The training plan consists of an integrated and creatively unique sequence of mentorship experiences to prepare the PI for their career. Training includes several opportunities to improve written and verbal communication, and professional development activities (course, conferences, seminars) for the PI to transition to a new research topic for an academic postdoctoral fellowship.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY / ABSTRACT This application “Restoring awareness of hypoglycemia in type 1 diabetes” proposes to elucidate the heterogeneity of impaired awareness of hypoglycemia (IAH) in type 1 diabetes through completion of a 24-month Sequential Multiple Assignment Randomized Trial (SMART) designed to better inform our understanding of the clinical and physiologic factors which contribute to restoration of counterregulatory defenses against hypoglycemia in response to educational, technologic, and pharmacologic interventions. Given the persistent barrier of hypoglycemia to the realization of achieving adequate glycemic control for most individuals with type 1 diabetes, there is a critical need to further understand the mechanisms contributing to hypoglycemia in type 1 diabetes in order to advance treatment approaches that may realize the benefits of near-normal glycemic control without the accompanying risk for severe hypoglycemia. The present proposal aims 1) to determine whether counterregulatory responses to a hyperinsulinemic hypoglycemic clamp can be restored in individuals with long standing type 1 diabetes and IAH using an adaptive randomized clinical trial design implementing state-of-the- art interventions including hypoglycemia avoidance education (standard-of-care; SOC), automated insulin delivery with hybrid closed loop technology (HCL), and novel mini-dose glucagon (MDG) pharmacology; 2) to determine the physiological factors associated with improved counterregulatory responses following intervention, including but not limited to age, diabetes duration, and continuous glucose monitoring (CGM) metrics; and 3) to validate a HypoA-Q short form questionnaire as a self-report measure for identifying IAH with measurement of counterregulatory responses derived from the hypoglycemic clamp. The proposed SMART study design will allow for rigorously determining both the degree of hypoglycemia avoidance necessary for improvement of counterregulatory epinephrine and autonomic symptom responses to insulin-induced hypoglycemia, as well as the additional factors such as age, diabetes duration, other CGM metrics, and validated IAH self-report that may predict individual responsiveness to SOC, HCL, and MDG interventions.

NIH Research Projects · FY 2025 · 2022-09

Abstract The aim of this grant is to identify and validate for therapeutic intervention, regulatory elements within the 5’ untranslated regions (UTR) of protein coding genes, known as upstream open reading frames (uORFs). In so doing, we aim to modulate the protein output from selected genes, offering a novel, translational approach with broad potential. To achieve the goals of our integrative MultiPI R01, we will leverage the expertise of both our labs spanning computational modeling, genomics, genetics, RNA biology and molecular biology. In Aim 1 we will combine large genomic and genetic datasets including gnomAD, the PennMedicine BioBank, and the UKBioBank to systematically identify functional uORFs and prioritize those for validation. The resulting database of human uORFs will be made publicly available and widely accessible as a user-friendly web-tool. Predicted regulatory uORFs suggested by the Aim 1 pipeline will be fed into the experimental Aim 2. In Aim 2 we will validate the high-priority uORF targets using luciferase assays. The results of Aims 2 will be fed back to the pipeline of Aim1 to improve prioritization of future uORFs. Overall, we expect the resources and discoveries made by this grant to shed light on the functional role of uORFs and offer therapeutic avenues for several hereditary diseases.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Signaling pathways have been implicated in a myriad of functions including mediating cell fate decisions, proliferation, migration, and spatial patterning, among other roles. Traditionally, these have been studied in specific cell types, at a limited number of timepoints, and in small numbers of cells. This presents three limitations: 1) Our understanding of these pathways has relied on “bulk analysis” of cell populations, which dilutes the signaling effects of individual cells. 2) Live imaging techniques preserve cellular resolution but lack the scalability to extend to whole tissues or organs. 3) Measurements at one or two timepoints do not capture the changing roles of these pathways at various developmental stages. Thus, we need to shift from small-scale “local” snapshots of signaling activities to large-scale “global” snapshots. Here I describe a novel technology that uses CRISPR/Cas tools to record signaling inputs in each cell’s genome and reads these signals at high throughput with single-cell RNA sequencing (scRNA-seq). As a proof-of-concept, I will apply this tool to investigate signaling events in the zebrafish brain. Several critical signaling pathways have been identified that influence neural progenitor fates and regulate spatial patterning of brain regions. In this study, I focus on the Notch and Fgf signaling pathways to test and validate the technology. The Notch pathway is an important mechanism that maintains the balance between neural progenitor cell proliferation and differentiation into neuronal cell types. Fgf signaling has multiple functions in the brain including forebrain development, spatial patterning and modulating left-right asymmetry. The first part of the proposal describes tools to record signaling activity in the zebrafish. It has 5 core components: 1) It uses 3 Cas orthologous proteins to independently record signals. 2) It has temporal inducibility of Cas activation. 3) It enables continuous signal recording at multiple timepoints. 4) It facilitates recording of multiple signaling inputs. 5) It can be scaled to whole tissues and organs and preserves the resolution of single cells. In the second part, I describe how the technology will provide new biological insights into Notch and Fgf signaling. I will investigate 1) whether Notch signaling in progenitors correlates with which types of neurons the progenitors differentiate into; 2) whether Notch-mediated asymmetric divisions reduced the cell heterogeneity within a progenitor niche; 3) how and when the activities of Notch and Fgf intersect to regulate neurogenesis. The combination of CRISPR/Cas-mediated signal barcoding and scRNA-seq provides a powerful platform for rapid, scalable and high-resolution investigation of signaling activity during development. I envision that this technology will provide a framework for innovation in tissue engineering, modeling disease and cancer, and studying adult regeneration.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT The goal of this proposal is to determine how epigenetic pathways regulate plasticity in social behaviors using the model ant Harpegnathos saltator. Specifically, we will test the hypothesis that external social cues are conveyed to chromatin by neuropeptides that regulate downstream transcription factors and associated epigenetic pathways to enable stable changes in social behavior. Epigenetic pathways are often disrupted in neurodevelopmental and behavioral disorders. Harpegnathos ants are an ideal model system to study brain epigenetics because workers and queens have the same genes but display distinct social behaviors. Furthermore, adult Harpegnathos workers can become queens via a remarkable phenotypic transition that involves plastic changes in reproduction, metabolism, and behavior. We have discovered that the ant homolog of the human gonadotropin-releasing hormone, the neuropeptide corazonin, is downregulated as Harpegnathos workers become queens and showed that it is necessary and sufficient to stimulate hunting in workers. Our preliminary data show that vasopressin, a neuropeptide with conserved social roles in mammals, is also preferentially expressed in worker brains, especially in those who do not express high levels of corazonin. In Aim 1, we will determine whether these two neuropeptides act on distinct or overlapping neuronal, molecular, and epigenetic pathways and whether they drive distinct social behaviors. Our previous work also revealed that Kr-h1, a transcription factor induced by corazonin, prevents unscheduled activation of “socially inappropriate” genes in the brain, thereby maintaining proper social behavior in both workers and gamergates. Preliminary studies identified another transcription factor, Fd3F, which is repressed by corazonin and might promote social plasticity in opposition to Kr-h1. Fd3F shares homology with pioneer transcription factors in other species, suggesting that an ability to reprogram chromatin states might underpin its function in behavioral reprogramming. In Aim 2, we will identify the changes on transcription and chromatin by which these transcription factors regulate brain and behavioral plasticity during adult caste transitions in Harpegnathos. The proposed experiments will leverage our previous experience with in vivo manipulation of gene expression in ant brains followed by behavioral and functional genomics analyses. Given that the neuropeptides, transcription factors, and epigenetic regulators investigated in this proposal are deeply conserved, our results should have broad impact on our understanding of how these molecular processes regulate social behavior.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT More than one million women undergo labor induction in the U.S. annually and over one-third of inductions end in cesarean delivery (CD). While many factors contribute to the CD rate, shifting the start of labor induction (e.g. cervical ripening) from an inpatient to outpatient setting is an intervention that can shape the early course of labor and significantly reduce the risk of CD. Possible mechanisms for the lower CD rate include improved sleep and relaxation at home, but also decreased time in the hospital and in turn, a decrease in medical interventions as well as a decreased exposure to variability in the diagnosis of abnormal labor progress. Based on published evidence that reduced time in the hospital is associated with reduced interventions, as well as early effectiveness data from small studies of outpatient Foley, we hypothesize that shifting the first part of the induction process from hospital to outpatient setting will be a promising and underutilized way to decrease CD rates. Furthermore, care delivery at home, in the outpatient setting, has been shown to improve healthcare utilization and cost by decreasing time in the hospital within internal medicine. The COVID-19 pandemic has further highlighted now as a critical time to think of novel and innovative ways to keep patients out of the hospital. This will be the first large, multicenter trial on outpatient cervical ripening powered to determine both effectiveness and safety of outpatient Foley. We propose a large (2300 women), multicenter, pragmatic, randomized trial to test the central hypothesis that outpatient cervical ripening with a Foley catheter will 1) decrease the primary CD rate and 2) reduce maternal and neonatal morbidity. Given the long time-lag between effectiveness studies and widespread implementation, we will additionally explore barriers and facilitators to implementation to enable rapid uptake and dissemination of our findings. Through a Type 1 hybrid effectiveness-implementation study, we will pursue the following specific aims: (1) Determine the effectiveness of outpatient Foley for cervical ripening in reducing the rate of CD among nulliparous women undergoing labor induction, (2) Determine the effectiveness of outpatient Foley in reducing maternal and neonatal morbidity and improving patient satisfaction, and (3) To characterize patient, provider, and organizational implementation determinants relevant to outpatient versus inpatient cervical ripening with a Foley catheter. Additional analyses will also compare maternal and neonatal resource utilization. A trial of this size and rigor is critical to changing the standard of care for millions of delivering women. Of critical importance, even if our primary effectiveness aim is negative, we will be powered to evaluate secondary effectiveness and safety outcomes, and the primary implementation aim will yield insight into ways to improve the induction process for thousands of patients through comparison of implementation determinants and resource utilization between the two arms.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Mid-substance Achilles tendinopathy is a painful, debilitating, and chronic tendon pathology. Rehabilitation exercises are the first treatment option for patients, but clinical studies have reported 20-60% of patients continue to experience pain and symptoms 5-years later. Despite these poor outcomes, rehabilitation protocols remain standardized because of insufficient evidence to support precision care. Our Parent R01 project will determine similarities in tendon loading and biomechanics in patients with mid-substance Achilles tendinopathy and how these similarities affect tendon healing, pain, and functional outcomes. In this Ancillary R01, we will leverage our innovative High-Density surface ElectroMyoGraphy (HDsEMG) interface to measure the plantar flexor neuromechanics during isolated ankle testing using dynamometry and functional tasks during walking on differing grades. Our preliminary data demonstrate that the relative contributions of the primary plantar flexor muscles can be modified by changing the knee angle during isolated and functional tasks. We will use this experimental model system to test the mechanistic link between plantar flexor neuromechanics, Achilles tendon loading biomechanics, and patient outcomes. In Aim 1, we will define neuromechanical profiles across Achilles tendinopathy disease progression. In Aim 2, we will link neuromechanical profiles with Achilles tendon loading profiles and stress imaging. By leveraging our large and well-characterized cohort of individuals with Achilles tendinopathy, this Ancillary R01 will establish how neuromechanical mechanisms (Ancillary R01) mediate complex biomechanical loading profiles of pathologic Achilles tendinopathy (Parent R01). This is a necessary next step to develop precision rehabilitation that factor patient-specific neuromechanics and Achilles tendon biomechanics. After successfully completing our proposed aims, we will test the clinical efficacy of precision neuromechanical rehabilitation in a randomized clinical trial.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Women constitute two-thirds of the Alzheimer’s disease (AD) population. While the sex-specific biological mechanisms underlying women’s increased prevalence are unclear, accumulating evidence points to menopause as a neurological transition state that may influence AD risk. Over the last quarter century, the vast majority of brain imaging studies have studied the neural basis of age-related cognitive decline in adults aged 65 and older. This convention overlooks one of the most significant neuroendocrine changes in a woman’s life— the transition to menopause—and leaves a gap in our understanding of the aging brain during the critical midlife years. The menopausal transition is marked by a sweeping decline in the production of sex hormones—up to 90% in the case of 17b-estradiol and progesterone. Animal studies provide powerful evidence that estradiol and progesterone play a neuroprotective role in brain regions vulnerable to neurodegeneration, including the prefrontal cortex and medial temporal lobes. However, the degree to which female reproductive aging leads to changes in human brain morphology, intrinsic brain network connectivity, and susceptibility to increased AD prevalence represents a significant knowledge gap that has yet to be adequately examined. This proposal will establish whether the decline in sex steroid hormones over the menopausal transition relates to vulnerability in brain circuits implicated in AD. In the F99 phase (Aim 1), I will probe the effects of reproductive aging on the brain in healthy women (N=90, ages 45–55), investigating the endocrine basis of neural aging in midlife. The well-characterized sample is enriched to include a balanced distribution of pre, peri, and post-menopausal women across a limited age range in order to isolate the effects of reproductive aging from chronological aging. I will first determine how the depletion of sex hormones in midlife alters large-scale functional brain networks using resting-state fMRI and computational approaches from complex systems analysis. I will then use high- resolution anatomical imaging of the hippocampus and surrounding medial temporal lobe to determine whether the depletion of sex hormones impacts specific hippocampal subfields (CA1-3, dentate gyrus, subiculum) and entorhinal, perirhinal, and parahippocampal cortices, regions enriched with sex hormone receptors. In the K00 phase (Aim 2), I will take the skills and insights gained from the F99 phase, including fundamental training in neuroendocrinology and brain imaging, and use them to establish the relationship between female reproductive aging and pathological AD biomarkers (b-amyloid, tau). To do this, I will leverage two large community cohorts (N~620, 60% female) that provide relevant hormonal, cognitive, and molecular positron emission tomography (PET) data from midlife subjects (ages 40-65) and build a neuroendocrine model of AD risk. Together, this proposal will identify the role menopause plays in contributing to female-specific vulnerability to Alzheimer’s disease, a severely understudied area in cognitive neuroscience with clear implications for women’s health.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY/ ABSTRACT In the midst of the COVID-19 pandemic, a parallel `infodemic,' an abundance of reliable information and inaccurate misinformation, persists. There has also been a significant increase in misinformation exchange and consumption, largely on social media platforms, which threatens individual and public health. An important challenge remains to develop strategies to detect trusted and accurate `signals' amidst dynamic misinformation `noise.' This misinformation contributes to confusion, distrust, and distress around health behaviors such as vaccination, mask wearing, and social distancing. The racial disparities in morbidity, mortality, social, and economic consequences of COVID-19 are well documented; less studied are variations in the information- seeking and COVID-19 health decision-making specific to Black and rural communities. Public health information and campaigns have traditionally relied on theory-based surveys or interview methods to measure knowledge and attitudes to design health messaging. Rapid expansion of social media use and parallel advances in machine learning analytics provide a unique opportunity to track public views, knowledge, and attitudes simultaneously to translate novel analytic insights into precision public health communication with an intentional lens on Black and rural communities. This proposal aims to build a computational framework to uncover heterogeneity in attitudes and misinformation exposure towards COVID- 19 vaccination, model predictors of highly engaging and persuasive messages (including sources, linguistic choices, and content); and to use pragmatic qualitative methods to understand individual response to social media misinformation with a specific lens on race (Black and white individuals) and location (rural and urban). While we focus our message development process on COVID-19 vaccination as a timely and critical behavior, and compare targeting across four specific audiences (Black rural residents, white rural residents, Black urban residents, and white rural residents), our approach is highly adaptable across health topics and scalable to a number of precision-targeted audiences. We see a need for flexible and nimble methods for rapid, human-centered content generation that supports accurate, equitable, and effective precision public health messaging. Computational tools powered by machine learning, predictive analytics, and natural language processing married with patient-centered qualitative methods offer a powerful synergy to conventional approaches to public health campaigns to identify and combat misinformation. The findings from this study will directly inform broader public health action and future strategies so that they can be deployed in the current pandemic and in ongoing efforts to address racial disparities in chronic diseases, HIV, cancer, maternal mortality, and mental health.

NIH Research Projects · FY 2025 · 2022-09

Summary There is a gap in the knowledge about how chondrocytes lose their phenotype and matrix production capacity during in vitro expansion. This gap in knowledge stems from the paucity of studies that directly interrogate chondrocyte genome architecture and transcriptional profiles in single cells to capture the inherent heterogeneity of cell differentiation. To fill this unmet gap, we will use state-of-the-art super-resolution imaging, single cell RNA Sequencing (RNA-Seq), high-throughput RNA-FISH (MERFISH) and metabolic labeling (FUNCAT) technologies to relate, on a cell-by-cell level, the chromatin nano-structure, transcriptional output, epigenetic modifications and matrix production capacity of single chondrocytes expanded in culture under different epigenetic and chemo-physical cues. We will further develop machine learning models to predict chondrocyte phenotype using super-resolution images of chondrocyte chromatin nano-structure. Our central hypothesis is that there are distinct chromatin nano-structural arrangements and transcriptional signatures associated with chondrocytes that have high matrix production capacity, and that chromatin nano-structure can be manipulated in a predictive manner via the combination of epigenetic and chemo-physical cues to improve chondrocyte therapeutic potential. The basis for this hypothesis is our preliminary super-resolution data of chromatin nano-structure in in vitro expanded chondrocytes and mesenchymal stem cells grown on substrates of varying stiffness and subjected to various chemical cues. The proposed work is significant as it will generate new knowledge about how chromatin nano-structure and epigenetic landscape regulates matrix production capacity of chondrocytes and how this capacity can be enhanced through manipulation of chromatin and epigenetic states. Our Aims are: matrix whether chromatin nano-structure and transcription are predictive of chondrocyte production capacity Aim 1: Determine Aim 2: Determine how chemo-physical and epigenetic cues impact transitions in chromatin nano- structure and matrix production in chondrocytes Aim 3: Determine whether predicted cues improve chondrocyte therapeutic efficacy In summary, we expect to contribute to the identification of new in vitro expansion conditions that maintain naïve chondrocyte phenotype and enhance their therapeutic potential. The proposed research is innovative as it represents a drastic departure from the status quo by applying multi-faceted, single-cell based imaging and sequencing technologies to determine the relationship between chondrocyte chromatin and epigenetic state, transcriptional activities, and matrix production. If successful, this work may change clinical practice by providing improved cell populations for cartilage repair.

NIH Research Projects · FY 2025 · 2022-09

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 emerged in China in late 2019, resulting in the COVID-19 pandemic. Like SARS-CoV (2002) and Middle East respiratory syndrome (MERS)-CoV (2012), SARS-CoV-2 can progress to cause lethal pneumonia. In contrast, infections with “common” respiratory CoVs (NL63, 229E, OC43) are largely limited to the upper respiratory tract. Furthermore, SARS-CoV-2 and in particular the omicron variant, can sometimes cause primarily upper respiratory infections. Thus, despite their highly conserved genome structure and shared replication schemes, human CoVs induce varying degrees of disease. Respiratory CoVs initiate infection through the nose, though few studies have addressed CoV infection of the nasal epithelium. We have an established cryobank of nasal epithelial cells from over 1000 genetically characterized individuals capable of being expanded and grown as air liquid interface (ALI) cultures, recapitulating the nasal respiratory epithelium. Our preliminary studies demonstrate that SARS-2 (and its emerging variants), MERS and NL63 all productively infect these cultures. However, NL63 only replicates at a lower temperature (33C), infects single cells rather than clusters (evinced by SARS-2/MERS) and causes a more cytopathic effect than SARS-2 or MERS, suggesting it may induce a robust local immune response thereby limiting its replication to the upper respiratory tract or stimulating an adaptive immune response prior to infecting the lower airway. One COVID-19 risk locus includes the leucine zipper transcription factor-like 1 gene (LZTFL1), which we show is highly expressed in ciliated nasal cells, with ubiquitous expression throughout the cytoplasm. Our preliminary data of SARS-CoV-2 infected cultures genotyped for the high vs low-risk LZTFL1 polymorphisms demonstrate that LZTFL1 could play a role in variability of SARS-CoV-2 spread. In addition, polymorphisms in OAS1, a sensor of double-stranded viral RNA that initiates the antiviral RNase L pathway, have been linked to COVID-19 resistance. We have extensive experience in this pathway and recently reported that SARS-CoV-2 activates RNase L while MERS-CoV shuts it down. Based on these and other data, we hypothesize that pathogenic outcomes of CoV infections are reflected in viral biology in the nasal epithelium. Thus, using a battery of diverse CoVs we will assess differences in cell entry and spread, optimal temperature for viral replication and shedding as well as host nasal cell responses to each CoV. We propose to use our biobank to identify host and viral factors affecting the establishment of infection, host cytokine and nasal antiviral responses and the contribution of polymorphisms in LZTFL1 and OAS1 genes in the outcome of infection. Our complementary expertise in coronavirus biology (Weiss) and nasal pathophysiology (Cohen) uniquely positions us to address these Aims. This work will contribute to understanding nasal CoV infection, the divergence of lethal and common CoVs as well as variation in clinical course among SARS-CoV-2 infections, and may lead to novel targeted prophylaxis or therapeutic strategies targeting the nose, the site of initial contact.

NIH Research Projects · FY 2025 · 2022-09

U01 Application: The role of senescent beta cells in T1D and T2D Abstract Recent studies including our own suggest a marked increase in β-cells expressing key components of cellular senescence in islets from Type 1 diabetes (T1D) and Type 2 diabetic (T2D) patients, implicating β-cell senescence as a critical contributor to islet dysfunction. Recently, it was reported that ablation of senescent cells by non-specific senolysis in mouse models of T1D and T2D improved diseease outcome. However, these studies did not determine which senescent cell type was relevant to the beneficial effect, nor did they address to what extent senescence occurs in the human endocrine pancreas before and during the development of T1D and T2D. To close this knowledge gap, in Specific Aim 1 we will determine the prevalence, transcription signatures and epigenomic landscapes of β-cell senescence in T1D, pre-T1D and T2D donors using immunostaining, imaging mass cytometry, single cell RNAseq, single cell ATACseq, DNA methylome determination and Cut-and-Tag analysis for key histone marks. In addition, we will evaluate the hypothesis that irreparable damage to telomeres drives senescence in β-cells, a possible scenario that could provide a mechanism for senescence to occur in islet cells of diabetic patients. In Specific Aim 2, we will test whether metabolic and/or inflammatory stressors drive senescence in human β-cells and determine the effect of senescence on β-cell function using scRNAseq and secretome analysis. We will evaluate if induction of senescence and the senescence-associated secretory phenotype (SASP) in human islet cells is p16 dependent, employing the pseudo-islets approach and using our hyperglycemic xeno-transplantation model to assess the direct effect of senolytics on human islet function. In Specific Aim 3, we will employ a novel transgenic mouse, the ‘SenKiller’ model, to enable cell-type specific and inducible ablation of senescent cells in any lineage including β-cells. Using this mouse model in combination with the appropriate β-cell specific Cre driver, we will provide a definitive answer to the question if senescent β-cells are critical in the development of glucose intolerance in models of T2D and islet autoimmunity in models of T1D. Together, this proposal will determine the occurrence of senescence among islet cells from T1D and T2D donors using large cohorts and multiple experimental modalities, explore the natural drivers of senescence and consequences to islet function as well as secretion of pro-inflammatory substances, and employ novel mouse models to unequivocally determine if elimination of senescent b-cells impacts diabetes progression in mouse models of T1D and T2D. The data generated here will address burning questions in the field, namely, is senescence increased in islets from diabetic patients, and are these cells important in the overall pathophysiology of T1D and T2D. These critical questions will have therapeutic relevance regarding the potential efficacy of targeting senescent β-cells with senolytic therapies.

NIH Research Projects · FY 2025 · 2022-09

Abstract/ Project Summary: Despite the widespread availability of effective antiretroviral therapy (ART), the HIV epidemic remains a persistent public health crisis in the United States. A significant proportion of individuals living with HIV are not consistently retained in care and do not achieve sustained viral suppression (VS), limiting both individual health outcomes and efforts to reduce community transmission. Longitudinal engagement in HIV care is needed for sustained VS and decreased community transmission of HIV. Although the outpatient setting is a vitally important aspect of care provision for PLWH, there are limited data on the impact of intra-organizational factors on HIV outcomes. The organizational social context (OSC) includes organizational culture (organizational norms and values that drive quality of care), organizational climate (perception of the culture and how it impacts personal well-being), and workers’ attitudes. Using a randomized controlled trial, we will implement ARC (Accessibility, Responsiveness, Continuity) to improve organizational behavior and HIV outcomes for PLWH. ARC is an evidence-based intervention that uses three strategies (ARC principles, ARC component tools, and ARC mental models) to create OSCs that support the implementation of interventions to improve patient outcomes. Clinics will be randomized to ARC (n = 2) or standard of care (SOC; n= 2). Those assigned to ARC will address factors occurring at the organizational level affecting care, including referral and treatment patterns for PLWH. A pre-implementation period will be followed by ARC and ARC-associated implementation strategies for 36 months and then a 12-month post-implementation period where we will continue to measure HIV outcomes in both arms. We will compare HIV outcomes, namely VS and retention in care, and intermediate outcomes, such as linkage to mental health treatment and staff turn-over in clinics assigned to ARC and OSC. We will also evaluate whether individual (self-efficacy, patient satisfaction, and provider trust) and organizational factors (OSC and cohesion of OSC measures) mediate the relationship between ARC, intermediate, and HIV outcomes. In preparation for the RCT, we will evaluate baseline OSC measures across 12 HIV clinics in Philadelphia and determine aspects of the OSC associated with VS and retention in care in a multi-level model adjusting for neighborhood and patient-level factors and clustering of patients nested in clinics and neighborhoods. We will then test the effectiveness of ARC in improving a primary outcome of VS and secondary outcome of retention in care at the end of the implementation period. We will examine the acceptability, sustainability, and cost of implementing ARC in outpatient HIV care. This research will advance understanding of the impact of organizational level factors on HIV treatment outcomes and health services research and the implementation of a disseminable evidence-based practice aimed at improving clinic culture and climate.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Individuals with Down syndrome (DS) have shortened lifespan and face severe challenges as they age. While the primary syndrome is caused by an extra copy of chromosome 21, and is present from birth, those with DS are at a much higher risk of developing sleep and metabolic disorders and more than two thirds of individuals experience cognitive decline that resembles early-onset Alzheimer's disease (AD). There is no unified mechanism or approach to treating these co-occurring conditions, which can lead to a drastic deterioration in quality of life. Published and preliminary data indicate that impaired proteostasis and aberrant activation of the proteostasis pathway, the unfolded protein response (UPR), is a biological mechanism common to AD, disrupted sleep and metabolic dysfunction. Moreover, protein folding stress can be a direct consequence of chromosome imbalance and data from humans and mice support the concept of aberrant UPR induction in DS. Thus, we propose that individuals with DS are susceptible to protein folding stress and that restoring proteostasis is a novel therapeutic approach to prevent co-occurring conditions. We posit that reducing proteostatic stress via chemical chaperone 4-phenyl butyrate (PBA) – a small saturated fatty acid that is an FDA- approved therapy for treatment of urea cycle disorders – will in turn ameliorate disturbances in sleep, metabolism, and cognition. Amyloid precursor protein (APP), a key player in familial AD, is one of the triplicated genes in DS. Overexpression and the consequent overproduction of amyloid beta (Aβ)-peptide leads to proteotoxicity that is instrumental in the early onset of AD neuropathology in the DS population. We have found that APP knockin (APPKI) mice treated with PBA display reduced proteotoxic stress and improved cognitive behavior, even when treatment was initiated after the onset of cognitive decline. Our data also indicate that PBA improves sleep quality in APPKI and normally aging mice. Moreover, several published studies indicate that PBA restores metabolic function in obese mice. Therefore, the global hypothesis of this proposal is that reduction of proteostatic stress with the FDA-approved small molecule chaperone PBA will ameliorate sleep, metabolic and behavioral deficits in a mouse model of DS. We will use the validated DS Ts65Dn mouse model that is trisomic for about two-thirds of the genes orthologous to human chromosome 21 and displays each of the relevant phenotypes to test whether PBA treatment rescues each of these co-occurring conditions. This proposal builds on a strong body of existing literature and new preliminary data supporting the potential of PBA, an FDA-approved therapy, for improving cognitive decline, sleep disturbances, and metabolic dysfunction. As these conditions co-occur in individuals with Down syndrome, this project represents a crucial first step towards developing a unified therapeutic approach.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The proposed study will examine insurance coverage for acupuncture therapy using both quantitative and qualitative methods, with the long-term goal of running interventional studies focused on insurance design that can improve access to safe and effective pain care. After decades of reliance on prescription opioids for chronic pain, the recent and widespread reduction in opioid prescribing is a welcome change. However, it is critically important that patients with chronic pain conditions like chronic low back pain (cLBP) have access to safe, effective, and affordable pain care. Acupuncture is an evidence-based treatment alternative for patients with cLBP, but its insurance coverage is inconsistent, making acupuncture cost prohibitive relative to other forms of pain care. Our first aim will measure trends in acupuncture use among a cohort of patients with cLBP using a national sample of claims data, then examine characteristics of patients who use acupuncture and identify other forms of pain care that they use in conjunction with acupuncture. We focus on patients with cLBP because it is one of the most common complaints cited by patients who engage in acupuncture; it is also the most common indication covered by insurers, including Medicare. In our second aim, we will evaluate the role of insurance design on acupuncture use, specifically the impact of cost sharing like copays, coinsurance, and deductibles, which have been shown to affect health care utilization at large and pain care in particular. A third aim will contextualize the claims-based findings with qualitative interviews with insurers and pain care providers, including acupuncturists, who will help us develop and refine an insurer-driven interventional study to encourage patients to engage in evidence-based pain care, like acupuncture therapy. While there are myriad ways to improve access to pain care, this project focuses on the role of insurance design and cost sharing. In terms of career development, this grant will support a training platform that will allow the candidate to reorient from behavioral health services research to acupuncture, pain care, and integrative health services research. An interdisciplinary mentorship team will oversee the following training goals: (1) Carefully review the research on the role of integrative medicine in comprehensive pain care, with a focus on acupuncture. (2) Develop an in- depth knowledge of the role of insurance coverage in pain care and identify empirical strategies to study how insurance design affects acupuncture use. (3) Learn how to conduct qualitative research that complements the candidate's strengths in health economics and claims-based research. (4) Build a knowledge base in interventional study design to propose a follow-up study that involves insurer-driven randomized trials that test whether alterations to insurance design affects the use of acupuncture. Through the training and research activities proposed in this K01, the candidate will become an expert in insurance coverage for acupuncture, gain a skillset in qualitative methods and interventional study design, and begin working toward her goal of translating research into actionable changes in policy or practice to support patients with chronic pain.