Johns Hopkins University

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract The goal of this study is to elicit preference-driven strategies to tailor contextually appropriate antiretroviral therapy (ART) packages, including long-acting injectables (LAI), for youth living with HIV (YLWH) in South Africa. Adolescents and young adults comprise 42% of all new infections in southern and eastern Africa, and among this population in South Africa, estimated ranges of viral suppression are from 10-50%. LAIs offer a potential solution for ART adherence and retention challenges faced by YLWH but may not work for all youth. We hypothesize that if guided by preferences and paired with appropriate implementation strategies, LAI can improve treatment outcomes among YLWH. Specific Aim 1: Guided by the Consolidated Framework for Implementation Research (CFIR), we will conduct 32 interviews with YLWH aged 12-29 years, parents, and pediatric and adult HIV providers (counselors, nurses, doctors) to identify context-informed bottlenecks to future implementation and opportunities for LAI delivery. Specific Aim 2: Evaluate acceptability, appropriateness and revealed preferences for LAI among YLWH 18-29 years in a partial patient preference trial pilot. YLWH will choose between treatment packages that include 8-weekly (2-monthly) LAI vs 2-monthly standard oral ART pills; those with no preference will be randomized (1:1) to receive LAI vs. pills. Active follow- up will continue for 6-months; in each arm, YLWH will be allowed to cross-over arms with clinical supervision between 8-16 weeks. The primary endpoints will be revealed ART preference (pills vs. LAI vs. no preference) at 0-months; and pill vs. LAI at 6-mos. Secondary clinical endpoints will include viral suppression (<50 copies per ml) and retention on ART; other secondary endpoints will include assessments of acceptability, feasibility and appropriateness of LAIs and adjacent strategies to retain YLWH in care. Specific Aim 3: Use preference and clinical outcomes data from Aim 2 along with other clinical trial and emerging observational data to model outcomes scenarios amongst YLWH. These aims will inform a fully powered R01 implementation trial, comparing viral load-informed, tailored LAI delivery and support strategies for YLWH using a sequential multiple assignment randomized trial (SMART) design.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT: This application seeks to establish a network for the investigation of gene-environment interaction in autism spectrum disorder (ASD) and outcomes among people with ASD. Much like the Psychiatric Genetics Consortium launched a unifying infrastructure for scaling genome-wide association studies in ASD, the Combining advances in Genomics and Environmental science to accelerate Actionable Research and practice in ASD (GEARs) Network effort will allow a centralized mechanism for GxE activities in ASD across multiple studies. Robust evaluation of GxE requires a large sample size, harmonized data on both genetics and the environment, and novel statistical methods for measuring and summarizing environments, genetics, and phenotypes. The GEARs Network seeks to compliment work in population studies with experimental models leveraging 3D brain organoids, reflecting multiple ASD-associated genetics backgrounds on which the impact of environmental risk can be evaluated on ASD-relevant neurophysiology endpoints. Finally, the GEARs Network will develop and implement a pipeline for outreach and dissemination of GxE findings. The successes of ASD genomics, emerging environmental evidence, and models of effective network collaborations for large-scale efforts make this the ideal time to create a GxE infrastructure for ASD research. Our team is uniquely poised to lead the GEARs Network, creating this opportunity at scale for the first time, and with a focus on both etiology and health outcomes among people with ASD. We have experience in leading multi-site collaborations, expertise in population and laboratory science, and required partnerships to foster communication across researchers and with the broader ASD community. This translational approach, informed by public health, will lead to improved understanding of both causes and consequences of ASD.

NIH Research Projects · FY 2024 · 2022-09

Summary/ Abstract Recurrent urinary tract infections (rUTI) are a significant problem among older women: 13% of female Medicare beneficiaries experience at least one UTI annually and >40% of these develop chronic recurrent UTI. Although UTIs are significantly reduced by vaginal estrogen therapy (VET), 50% of those using VET continue to experience UTI recurrences. It is unknown why some women benefit from VET while others do not. This application focuses on interrogating two mechanisms likely to be central to the effectiveness of VET. The first is the urogenital microbiota: an increase in vaginal lactobacilli is the purported mechanism by which VET reduces rUTI. However, recent studies suggest that not all lactobacilli are equally beneficial: vaginal microbiota dominated by L. crispatus may be more protective (possibly via the production of D-lactic acid, which inhibits E. coli growth). Important and unanswered questions include how VET influences specific Lactobacillus spp., whether changes to specific Lactobacillus spp are the key to successful prophylaxis, and how VET affects the urinary microbiota, which may play a critical role in UTI susceptibility. A second mechanism addressed by this application is the host vaginal and urinary immune response. Estrogen appears to influence localized urogenital immune responses, including Th17 and Th1 versus Th2 pathway signaling. Animal studies suggest that these compartmentalized immune responses play a critical role in UTI susceptibility, but human data are lacking. This application will address these unanswered questions. Postmenopausal women with rUTI will be treated with VET. Samples collected before and after VET will characterize vaginal and urinary microbiota (16S rRNA gene sequencing), soluble mediators of inflammation in both compartments, and vaginal D-lactic acid. Aims 1 and 2 of this proposal will investigate the impact of VET on the urogenital microbiota and urogenital immune responses, respectively. Aim 3 will characterize the urogenital environments of participants who continue to experience rUTI during VET versus those who remain UTI-free. The accomplishment of these aims will provide pilot data for a larger and more definitive clinical trial. Thus, this application is responsive to program announcement PAS-20-160, which supports small clinical trials to provide critical preliminary data. This proposed research will provide data needed to plan a rigorous, adequately powered trial to identify the characteristics associated with successful rUTI prevention. These proposed studies are a key step toward our goals of identifying biomarkers that reliably predict a successful response to rUTI prophylaxis and ascertaining the biological conditions required for successful UTI prevention. Ultimately, an understanding of the mechanisms of rUTI prevention will allow the development of novel and effective prevention strategies for postmenopausal women suffering from rUTI.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY This year alone, over 600,000 people in the U.S. will die from cancer, with each patient losing an average of 15.6 years of life. However, upwards of 25% of these deaths could likely be avoided if these cancers were detected at earlier stages. One particularly attractive approach for cancer diagnostics is the use of circulating cell-free DNA (cfDNA) from so-called “liquid-biopsies” of patient-derived serum/plasma as these samples are often enriched in genetic material from tissues, including tumors, located throughout the body. Nonetheless, tumor specific alterations, such as mutations and aberrant DNA methylation, are typically only present at extraordinarily low copy numbers (< 10 copies/ml) and fractional concentrations (< 0.1%) within a large background of healthy-tissue DNA. This issue in particular has proven problematic for current technologies and has thus far precluded development of a cfDNA diagnostic method that is simple, low-cost and, most importantly, able to detect cancer at stages sufficiently early to improve patient outcomes. In the present project, we aim to develop REM-DREAMing: a low-cost, highly-multiplexed digital methylation analysis platform that provides highly-sensitive and parallelized assessment of cfDNA methylation patterns to enable detection of rare tumor DNA, even from early-stage cancers. At the core of the REM-DREAMing platform is a unique, locus-specific DNA methylation assay, called DREAMing (Discrimination of Rare EpiAlleles by Melt), that has been successfully developed by our lab to provide detection and absolute quantification of cancer-specific DNA methylation even at extremely low fractions (<< 0.1%). Recently, we successfully incorporated the DREAMing assay into a massively-parallel digital microfluidic array to enable detection of a single copy of aberrantly-methylated DNA in a background of 2 million unmethylated alleles. Here, we propose to dramatically enhance the microfluidic DREAMing approach by significantly expanding its digitization power and incorporating novel, methylation-agnostic probes with a unique ratiometric fluorescence multiplexing scheme to achieve simultaneous digital assessment of a panel of 50 “cancer-detecting” and “cancer-identifying” methylation biomarkers, enabling liquid-biopsy-based detection and identification of early-stage cancers at a cost of only a few dollars per sample. To achieve this goal, we plan to accomplish the following aims: (1) Develop dual, 27-plex DREAMing assay panels targeting a panel of 50 pan-cancer-detecting and cancer-identifying methylation biomarkers. (2) Design, fabricate and validate a dual 400k-well, 4-color fluorescence-decoding dHRM platform to perform parallelized REM-DREAMing for simultaneous detection and identification of 50 methylation biomarkers. and (3) Assess and benchmark the ability of the REM-DREAMing platform to detect and identify six different cancer types from liquid biopsies.

NIH Research Projects · FY 2025 · 2022-09

Fifty percent of neonatal encephalopathy cases result from hypoxic-ischemic encephalopathy (HIE). Therapeutic hypothermia (TH), the only approved therapy remains state of the art care for HIE, despite only a 30% reduction in death and significant disability. Our limited ability to accurately track TH efficacy limits identification of babies, who may benefit from adjunctive therapies. Under R01HD086058, our team enrolled neonates with HIE treated with TH and tested whether circulating brain injury biomarkers used in adults were associated with HIE severity, MRI and 2-year outcomes. We identified the novel biomarkers significantly associated with the proposed outcomes and published 22 peer-reviewed original, high-impact manuscripts. Our team has extensive experience in biomarkers in children (1R01HL150070), brain injury biomarkers in HIE (U01 NS114144) and real-time machine learning integrating within health systems (R61HD105591). Our central hypothesis is that a holistic and integrative approach, including deep clinical and community-based data, and molecular biomarkers of multiple biologic pathways, analyzed using a fully connected parsimonious neural network will best describe relationships with longitudinal outcomes, and be able to predict response to TH in individual patients. Our outstanding group of investigators from Johns Hopkins University, University of Virginia and University of Alabama Birmingham, propose the following Aims: Aim 1a. Perform clinical data- driven modeling to ascertain TH effectiveness. We will use deep phenotyping data sets of all maternal, neonatal, community-based, and follow-up data collected retrospectively (2016-2021) and prospectively thru year 1, from neonates treated with TH at the 3 centers (n = 500) to model TH efficacy using multivariable methods against longitudinal outcomes. Aim 1b. Identify novel molecular signatures for HIE insult severity which predict response to TH. Using our discovery (N=178) TH treated HIE cohort, we will determine if circulating brain injury proteins, metabolites and exosome proteins and nucleic acids are associated with TH efficacy. Aim 1c. Determine relationships emerging from integration between clinical, community-based, and molecular markers using a fully connected parsimonious neural network approach. 1C.1 Use computational simulations to identify the levers, modifiable risk factors and interventions associated with the probability of negative outcomes, in the neural network, and 1C.2 Determine in silico whether optimization of the neural network using those levers at the individual patient level, results in a reduction in the predicted probability of negative outcomes. Aim 2. External validation of neural network and estimation of potential clinical gain achievable by optimization of the neural network, in prospective patients (Years 2-5). Completion of our aims will identify the clinical, socioeconomic, and molecular mechanisms driving clinical heterogeneity in HIE and response to TH. We will then be poised to rapidly deploy a dynamic, precision-based model to optimized patient selection for future HIE adjunctive therapies.

NIH Research Projects · FY 2025 · 2022-09

Hypertensive disorders of pregnancy, such as preeclampsia (PEC) cause significant maternal morbidity and mortality, and have emerged as an important early life, and potentially modifiable, risk factor of heart disease in younger women. PEC is diagnosed as new onset hypertension after 20 weeks gestation with evidence of end organ damage, thought to occur because of placental ischemia and an imbalance of angiogenesis. Importantly, PEC is associated with significantly increased CVD risk that persists into later life, far beyond the pregnancy period. Understanding how PEC affects cardiovascular (CV) health both from a biological and behavioral perspective is critical to inform risk communication, stratification and to identify and target new treatments to reduce CV disease (CVD), especially among young, high risk women. Although patients with PEC often have traditional pre-pregnancy CV risk factors such as type 2 diabetes and obesity, these risk factors do not fully explain the elevated CV risk conferred after a pregnancy complicated by PEC. Two factors thought to contribute to the pathogenesis of PEC and injure endothelial cells are an imbalance of angiogenesis and alterations in renin-angiotensin-system (RAS), driven by placental ischemia and resulting in oxidative stress with systemic inflammation. From a socio-behavioral perspective, there is evidence that social determinants of health and psychological stress influence PEC severity and future CVD outcomes. Women’s lived experience, including how they experience stress and neighborhood factors affect health outcomes significantly in PEC. We recently developed noninvasive, reproducible MRI-based methods to measure coronary endothelial function (CEF), offering a means to probe mechanisms contributing to coronary artery disease (CAD) pathophysiology in postpartum women with recent PEC. Abnormal CEF plays a critical role in the development, progression and clinical manifestations of CAD, independently predicts CV events, and is a target for medical interventions. We propose in this application to determine in postpartum women with PEC: 1) whether CEF, and markers of inflammation/imbalanced angiogenesis are inversely related and 2) whether measures of stress (both psychosocial stress and physiologic measures) are associated with endothelial abnormalities and postpartum hypertension. We will determine, in women with a history of PEC, whether reduced CEF can be explained, at least in part, by increased inflammation/abnormal angiogenesis/stress. Together these studies will offer new pathophysiologic insights into increased CVD risk in women with PEC, and which factors contribute most to vascular dysfunction. This study, if funded, will address significant knowledge gaps regarding sex specific variables that affect postpartum vascular health in young women and provide data to design future interventional studies to improve health outcomes in PEC.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Access to hearing care is unequal with large disparities among older adults. Despite evidence that supports community health worker (CHW) care models as an approach to health disparities, community-delivered hearing care models are only beginning. Among existing efforts, only the HEARS (Hearing Health Equity through Accessible Research & Solutions) intervention is specifically designed for older adults and includes the provision of over-the-counter (OTC) hearing technology in a community setting through a theory-driven approach. The HEARS intervention was designed and tested with NIDCD support from 2015 to 2020 (R22/R33DC015062) to address common barriers to hearing care experienced by older adults, particularly racial/ethnic minority and low-income older adults. Preliminary data from the randomized controlled trial demonstrate the efficacy of HEARS as a first-in-kind intervention delivered entirely in the community by older adult peer mentors using OTC hearing technology with outcomes on par to hearing aids fit by audiologists. With increasing availability of OTC devices and a growing demand for new care models, the HEARS intervention is poised to be brought to scale. To move HEARS into practice, we propose partnering with local Area Agencies on Aging (AAAs), a national network of public and private non-profit agencies who provide services to older adults aging in place. This proposal will complete the HEARS intervention package and test the delivery of the HEARS intervention entirely within a community setting by community-based audiologists working with older adult peer mentors in AAAs, positioning the HEARS program for broader implementation through AAAs. The proposal has the following aims: R21 Aim 1 To develop the train-the-trainer curriculum and manuals for audiologist supervisors that reflect the needs of audiologists and ensure the safety, efficacy, and fidelity of a peer mentor-delivered intervention as guided by the Scientific and Community Advisory Boards. R33 Aim 1 To recruit and train an audiologist-peer mentor care team of 2 audiologists and 9 peer mentors (3 per site) through partner AAAs (n=3) to assess the trainer curriculum. R33 Aim 2 To conduct a community- based pragmatic trial of delivering the HEARS intervention through 3 AAAs throughout Maryland via a within- subject 3-month intervention trial (n=100 per AAA site). R33 Aim 3 To develop an implementation strategy for the HEARS intervention as delivered through AAAs via a mixed methods approach based on the barriers and enablers encountered in the pragmatic trial. This proposal brings together a multidisciplinary team of investigators in a highly collaborative effort spanning academia, government, and non-profit organizations. This proposal will rigorously extend evidence for a community-delivered hearing care that leverages OTC devices and CHW-partnered care and will directly inform the implementation and scaleup of the HEARS intervention.

NIH Research Projects · FY 2025 · 2022-09

Neurosarcoidosis (NS) represents the neurologic manifestations of sarcoidosis, a multisystemic granulomatous inflammatory disorder of unknown cause. NS may be observed in 5-15% of patients with sarcoidosis, a worldwide disease that disproportionally impacts African Americans and whites of northern European heritage. Our preliminary studies showed NS has a wide spectrum of clinical phenotypes that includes meningitis, encephalitis, and myelitis. We also found that cerebrospinal fluid (CSF) from patients with NS reveal a unique profile of immune mediators frequently associated with infections (interferon-γ, tumor necrosis factor-α and interleukin-6) and antibody signatures linked to Mycobacteria antigens. Based upon these observations, we hypothesize that the pathogenesis of NS is due to a neuro-inflammatory response to antigens derived from exposure to infective agents in susceptible individuals with the clinical phenotype determined by specific gene expression signatures. This study engages two centers with existing cohorts of NS patients with prospective collection of clinical data and biological samples to dissect CSF immunopathogenic pathways, define immune profiles, and uncover antigens or pathogens which may be associated with NS phenotypes. Our specific aims focus on associating clinical NS phenotypes with immune profiles and gene expression pathway signatures in CSF and the link with host or pathogen-associated antibodies. In Aim 1, we will perform rigorous phenotyping of NS patients, and use biological samples such as CSF to characterize previously identified cytokine and acute phase reactants and their usefulness as biomarkers of disease outcome. In Aim 2, we will use host CSF transcriptional profiling to identify specific molecular signatures and pathways present in NS will establish immunopathogenic mechanisms and factors that contribute to dynamic neuroinflammation and disease progression. In Aim 3, we will use state of the art phase display libraries and phage-displayed immunoprecipitation sequencing techniques to determine the presence of antibodies to host and microbial- associated antigens which may identify triggering mechanisms related to the NS inflammatory process. All aims are well integrated as Aim 1 will provide a well characterized and phenotyped cohort of patients with NS which would facilitate a more precise identification of disease pathways in the CSF transcriptomic analysis outlined in aim 2, and host- or pathogen-related antibody response discovery in Aim 3. The studies proposed will address critical voids in our understanding of the pathogenesis of NS and suggest future novel therapeutic strategies.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This five-year K01 Mentored Research Scientist Development Award will provide Dr. Rucinski with the mentorship and training to become an independent investigator focused on research that optimizes the health and well-being of adolescents and young adults (AYA) at high risk of HIV acquisition and transmission in the United States (US). Training is at the foundation of this proposal and comprises mentorship with expert and multi-disciplinary HIV and implementation science (IS) researchers, didactic coursework, experiential learning, meetings with key stakeholders and community leaders, and rigorous mentored research. The following training objectives will be accomplished during this award: (1) Obtain scientific expertise in the concepts, theories, tools and methods used to advance HIV-related IS research and practice; (2) Develop comprehensive skills in qualitative research methods to support the application of mixed-methods approaches for HIV-related IS research; and (3) Acquire content expertise in adolescent health and development to inform the tailoring, adaptation, and testing of developmentally appropriate HIV services for AYA. Supported by these objectives, the proposed mentored research project will generate a tailored packaged of implementation strategies to strengthen contextually appropriate antiretroviral therapy (ART) programs for AYA aged 15-24 living with HIV in the US. In the US, 19% of all new annual HIV infections are among AYA. Youth living with HIV are less likely to achieve viral suppression compared to adults, with cohort data suggesting that as little as 12% of AYA new to care are able to achieve viral suppression within 6 months. The barriers AYA experience in accessing treatment in the US reflect broader global challenges documented in high-incidence settings, such as in sub-Saharan Africa. These barriers include mental health concerns, economic insecurity, and a lack of social support. Lessons from these settings demonstrate that models such as dedicated weekend clinic hours, peer mentorship, and other psychosocial services can support ART adherence and viral suppression for AYA living with HIV. Given the rapidly evolving landscape of HIV treatment in the US, including through innovations such as long-acting injectables, there is an urgent need to develop strategies that support youth living with HIV to initiate and continue on ART, to conduct this research in a way that is efficient, and to apply global programmatic lessons to the US context with the overall goal of optimizing adolescent health. Guided by the Consolidated Framework for Implementation Research, we propose the following aims: (1) Examine the social, structural, and clinic-level factors associated with retention in care and HIV viral suppression among AYA who were newly infected or diagnosed with HIV; (2) Refine a tailored package of implementation strategies to optimize HIV treatment for AYA living with HIV; and (3) Evaluate the preliminary effectiveness of a combined psychosocial/structural intervention in improving HIV viral suppression for AYA living with HIV. Findings will form the foundation for a R01-level randomized hybrid effectiveness trial and will facilitate Dr. Rucinski’s transition to independence.

NIH Research Projects · FY 2024 · 2022-09

PROPOSAL SUMMARY Adverse childhood experiences (ACEs) are childhood exposures to potentially traumatic family and community experiences that can have lasting, negative impact on health, are a driver of health disparities, and have potential intergenerational transmission of effects. Preliminary data from the parent study for this proposed study indicate that ACEs are highly prevalent in low-income, urban parents, with 54.7% of 139 parent participants reporting 4 or more ACEs. Positive childhood experiences (PCEs) are relational experiences that confer a sense of safety and nurturance during childhood, such as feeling safe with at least one adult in the home. PCEs are associated with improved physical and mental health. PCEs have the potential to promote positive outcomes even when ACEs are high. Current neighborhood violence is expected to compound the impact of ACEs on parenting practices due to ongoing or recurrent activation of neurobehavioral responses to threat. The purpose of this convergent mixed methods study is to understand how parents' exposures to ACEs and PCEs impact their parenting practices, particularly in the context of living in unsafe neighborhoods by addressing the following specific aims: 1: Test the associations among parents' ACEs, PCEs, and more positive parenting practices in a sample of parents (n=200) raising young children (2-8 years old) in Baltimore. 2: Examine contextual effects of current neighborhood safety (violent crime rate and parent perception) by testing the interactions between threat-type ACEs and neighborhood safety variables, controlling for other neighborhood-level SES characteristics. 3: Understand how parents' own childhood experiences (ACEs and PCEs) influence their parenting practices in the context of low versus moderate to high neighborhood safety. The proposed study will be nested within an ongoing parent study in Baltimore City Public Schools. The proposed study will use baseline data only for a cross-sectional quantitative arm, followed by a qualitative arm that integrates with the quantitative data. Multivariate regression, multilevel modeling, and conditional process modeling will be used to test associations among ACEs, PCEs, and parenting, and then to test these associations and the association with neighborhood safety. In the qualitative phase, indvidual interviews will use an interpretive phenomenological approach to gain a better understanding of the processes and contextual factors that contribute to participants' parenting in the context of neighborhood safety. This design integrates the analysis of associations between ACEs, PCEs, parenting practices, and neighborhood safety with analysis of parents' perspectives on the impact of their childhood experiences on parenting, role of neighborhood safety, and other contextual variables. This research will identify protective factors in the intergenerational transmission of ACEs and uses geospatial data to explore how neighborhoods influence the impact of ACEs and PCEs. It will inform future research on family- and neighborhood-level interventions to promote safe, stable, nurturing relationships in families and neighborhoods to reduce the rate and impact of ACEs.

NIH Research Projects · FY 2025 · 2022-09

Title: Protein assemblies as genetically encoded mechanical actuators for intracellular mechanobiology research Project Summary Cells are continuously subjected to mechanical cues that regulate diverse biochemical and biophysical processes. In the rapidly growing field of mechanobiology, various methods, including but not limited to substrate engineering, optical/magnetic tweezers, atomic force microscopy, pipette aspiration, and microfluidics, have been developed and exploited for mechanical manipulation of live cells. Though with grand success, these paradigms primarily apply mechanical forces at the cellular surface, while direct intracellular perturbation remains underexplored. The limited capability of intracellular force exertion impedes in-depth investigation of critical fundamental questions such as how forces are translated inside the cells and regulate the output functions. Therefore, we plan to fill the technological gap by developing a toolbox of genetically encoded peptides/proteins as intracellular mechanical actuators. We will rationally design and engineer peptides/proteins that can spontaneously form in-cellulo nanoscopic or microscopic assemblies with various sizes, shapes, surface chemistries, and mechanical properties to mimic intracellular mechanical milieu. The corresponding biological responses of cells will be probed by molecular sensors and optical microscopy. With the assistance of numerical simulation, the mechanical interactions between protein assemblies and subcellular structures will be recapitulated and correlated to the change of biological processes. The tools, once developed, will be exploited to study mechanoresponses of membrane receptors and cell nuclei. The genetically encoded mechanical actuators can afford several distinct advantages: 1) It allows direct, chronic, and precise exertion of force intracellularly; 2) A large number of cells can be transformed and characterized simultaneously, enabling high-throughput probing and analysis; 3) The genetic delivery of the tool and the contact-free perturbation makes it readily applicable to more complex and physiologically relevant biological systems, such as organoids, ex-vivo tissues, and even the live animal models. We believe the new genetically encodable tools can shift the paradigm for intracellular mechanobiology research and help advance our understanding of organelle mechanosensing,

NIH Research Projects · FY 2025 · 2022-09

SUMMARY A growing number of serious disorders ranging from syndromic autism and intellectual disability to cancers of the brain and gut have been linked to intracellular members of a family of electroneutral Na+/H+ exchangers, including endosomal isoforms NHE6 and NHE9 (eNHE), that regulate pH and Na+ within the compartments of the endo-lysosomal pathway. Plasma membrane NHE isoforms have been thoroughly characterized and pharmaceutically targeted. In contrast, intracellular NHE remain poorly studied due to limitations and challenges in sensing organelle-specific lumenal ions. Furthermore, overlapping distributions of eNHE isoforms and contradictory reports on the direction of sodium and proton transport within organelles has hindered a mechanistic understanding of transporter function and physiology. Case reports linking disease to eNHE genetic variants are sporadic and genotype-phenotype correlations are incomplete. This proposal brings together three research groups with unique and complementary expertise, together with powerful tools and resources to tackle these problems. To overcome the technical challenges in measuring the activity of these transporters, we have developed a multi-functional fluorescent reporter for both Na+ and H+ to precisely assay intracellular Na+/H+ exchange. This reporter can be targeted to specific organelles to simultaneously read out Na+ and H+ levels therein using an imaging method called two-ion measurement. In Aim 1, we will deploy this reporter to specific compartments along the endo-lysosomal pathway to quantify [Na+] and [H+] in both healthy and disease states. We will determine the functional contribution and mode of transport of individual eNHE isoforms in key organelles. This aim will lay the groundwork for functional analysis of clinically impactful gene variants in eNHE. To capture the disease landscape for eNHE, in Aim 2 we will evaluate the clinical significance of rare and common gene variants in SLC9A6 and SLC9A9. For these analyses, we will leverage large-scale exome sequencing of a clinical cohort, paired with their de-identified electronic health records. Combining genetic associations, gene expression and functional analysis will provide mechanistic insight on the biological basis of disease associated with eNHE. In summary, our comprehensive biochemical mapping of the endo-lysosomal pathway and disease-agnostic approach to link gene variants and expression to phenotypes will capture a broad range of cellular and clinical correlates that will pave the way to successful therapeutic targeting of these transporters in disease.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT Critical care is the most complex, resource-intensive and costly care setting contributing over $130 billion to national health expenditures annually. Approximately 4 million patients are admitted to intensive care units (ICUs) each year with average mortality rate ranging from 8-19. Recently COVID-19 has highlighted that staffing interprofessional teams in ICUs is quite challenging. One feature of healthcare staffing that has received much attention in settings outside of the ICU is continuity of care – whereby a patient is cared for by a small team of identified professionals over time – has been long recognized as an essential attribute of high- quality, patient-centered care. In primary care, greater continuity of patient care is associated with fewer emergency department visits and hospitalizations, lower healthcare costs, and higher patient satisfaction. Continuity-based acute care staffing models also exist, but are rarely adopted and sustained in practice, particularly in ICUs where continuity-based assignments can be challenging to operationalize. Importantly, although critical care is delivered by interprofessional teams of physicians, nurses, and respiratory therapists, continuity of interprofessional ICU teams has never been conceptualized or measured before. Without this knowledge, it is difficult to know whether hospitals and administrators should prioritize continuity of ICU care, which could be a missed opportunity to improve quality of patient care and outcomes in this critically ill, costly patient population. The overall goal of our study is to examine the effect of continuity of ICU interprofessional teams on patient outcomes and organizational economic outcomes, in order to develop an interprofessional assignment decision-support tool that optimizes the continuity of interprofessional ICU team care. We propose to examine the quality and costs of an interprofessional team continuity staffing approach in ICUs to guide future interventions in 13 ICUs across two different healthcare systems using data from over 2 years. We will examine interprofessional team continuity of shift-level ICU clinician teams (a nurse, a physician, and a respiratory therapist) assigned to each patient during the ICU stay. We define two dimensions of team continuity: intra-professional continuity, measuring each of the clinician’s experience caring for the patient previously during the patient’s stay, and inter-professional continuity, measuring the clinicians’ joint experience working together as a team. We hypothesize that care delivered by shift-level ICU teams with higher intra- and inter-professional continuity will result in improved patient outcomes and reduced costs. Our research team has a strong record of joint publications on attributes of patient care teams including continuity of care with expertise in health economics, critical care nursing and medicine and engineering. Completion of the study will generate the most robust evidence, to date, to inform organizational priorities about continuity-based interprofessional staffing in ICUs, to improve care for critically ill ICU patients.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Infertility – the inability to conceive a pregnancy within 1-2 years – causes intense psychological distress and other adverse outcomes for millions of women and men but remains understudied in low- and middle-income countries. Infertility research is particularly limited in sub-Saharan Africa (SSA), despite findings suggesting SSA has among the highest levels of infertility globally, especially secondary infertility (the inability to conceive a second or higher-order pregnancy within 1-2 years). Infertility estimates in SSA range widely from 2-31% de- pending on location, population, definitions, and assessment method; the broad range of estimates highlights the need for further research to evaluate model assumptions. Besides uncertainty about the extent of infertility, we lack an understanding of the mechanisms contributing to adverse outcomes. Women experiencing the “ag- ony of infertility” are often blamed, causing physical and emotional abuse, marriage instability, poverty, social isolation, psychological distress, and even suicide. These impacts may be exacerbated by fears of infertility and perceived community-level infertility stigma, which have negative impacts on women’s health, health be- haviors, and care seeking. However, existing research on these phenomena are limited. The objective of this training grant is to conduct mentored research to address these knowledge gaps by improving the field’s un- derstanding of infertility levels, stigma, and fears in SSA. This work leverages Performance Monitoring for Ac- tion (PMA) in Uganda, an ongoing, longitudinal study of reproductive aged (15-49) women. I will work with my mentors to achieve three specific aims: 1) Improve population-level infertility prevalence estimation by evaluat- ing infertility measurement model assumptions in Uganda; 2) Understand Ugandan women’s and men’s per- ceptions, fears, and experiences of infertility and its related stigma, and; 3) Identify dimensions of infertility stigma and fear among women in Uganda. Results have implications for both population-based measurement of infertility prevalence and our understanding of infertility stigma and fear, which can inform interventions to reduce them and their impacts on health and well-being. The corresponding training objectives that will enable me to complete the proposed research include various training activities and mentorship in: 1) advanced ana- lytic techniques for the measurement of infertility; 2) qualitative methods; 3) psychometric research techniques; 4) infertility etiologies and relevant conceptual frameworks, and; 5) NIH-grant writing. The proposed research will lay the groundwork for submitting a competitive R01 application in year 4 proposing a mixed-methods lon- gitudinal study of infertility, related stigma and fear, and their impacts in SSA. The complementary research and training will move me toward my long-term goal to become an independent investigator using interdiscipli- nary, mixed-methods approaches to improve the scientific understanding of health disparities in fertility-related behaviors (both fertility prevention and promotion) and outcomes in low-resource settings.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract Elongating ribosomes frequently encounter obstacles that cause ribosomes to stall. Stalled ribosomes are then targeted by rescue factors to recycle the ribosomal subunits and target the faulty mRNA and nascent peptide for decay. This process is critical for cellular fitness in bacteria: its loss often results in decreased pathogenicity, viability, or antibiotic resistance. In E. coli, ribosome stalling leads to collisions, which recruit the endonuclease SmrB to cleave the mRNA between collided ribosomes. These ribosomes are then targeted by the rescue factor tmRNA. Several lines of evidence suggest E. coli employ a second pathway to split antibiotic- inhibited ribosomes into subunits; however, a ribosome-splitting factor has yet to be identified. The DExH-box RNA helicase HrpA decreases E. coli sensitivity to ribosome-targeting antibiotics, suggesting this protein has a ribosome-associated function in antibiotic resistance. In fact, HrpA has been shown to rescue ribosomes stalled on reporter mRNA and resolve global antibiotic-induced ribosome collisions, indicating that it plays a role in clearing stacked ribosomes during translational stress. The proposed experiments will explore whether HrpA is a novel ribosome rescue factor that splits stalled ribosomes. Because ribosome collisions are critical for ribosome rescue in yeast, mammalian cells, and E. coli, Aim 1 will determine if ribosome collisions also recruit HrpA. Protein readout from a reporter-based assay in wild-type and ΔhrpA cells will be used to test if HrpA rescues collided ribosomes from mRNA. To determine if HrpA preferentially associates with collided ribosomes in vivo, sucrose density gradients will be used to determine if HrpA sediments with ribosome subunits, single ribosomes, or collided polysomes from cell lysates that are treated with ribosome-stalling antibiotics. Finally, the impact of HrpA on ribosome position and ribosome collisions transcriptome-wide will be explored using ribosome profiling in antibiotic-treated wild-type and ΔhrpA cells. Two S. cerevisiae RNA helicases in the same DExH-box family as HrpA are involved in ribosome splitting; therefore, Aim 2 will determine if HrpA similarly splits stalled ribosomes in E. coli. Because the protein Hsp15 preferentially associates with 50S ribosomal subunits from prematurely split ribosomes, HrpA splitting activity will be assayed in vivo by examining Hsp15 sedimentation in sucrose density gradients in antibiotic-treated wild-type and ΔhrpA cells. Finally, HrpA will be tested for its ability to split stalled ribosomes into subunits using an in vitro biochemical assay with purified components analyzed in sucrose density gradients. This assay will be used to determine ATPase activity and precise substrate recognition of HrpA. Characterization of HrpA will provide valuable insight into how E. coli mitigate the effects of ribosome-targeting antibiotics, marking it as a promising potential target of antimicrobial compounds in the treatment of human disease.

NIH Research Projects · FY 2025 · 2022-09

Project Abstract Aphasia is a devastating complication of stroke. Speech and language treatment (SLT) can be helpful in restoring language function, but not all individuals show improvement. Recent studies indicate that Transcranial Direct Current Stimulation (tDCS) is a promising adjuvant approach to enhance the effectiveness of SLT. tDCS is a noninvasive, non-painful, electrical stimulation of the brain. It is believed that tDCS boosts neural plasticity that underlies recovery with SLT. A majority of the tDCS studies of aphasia have stimulated the left hemisphere regions. However, left hemisphere lesions common in post-stroke aphasia affect the electrical field in unpredictable ways, potentially preventing stimulation from reaching perilesional tissue associated with optimal recovery. Our work addressed this problem by stimulating a novel region, the right cerebellum. The right cerebellum is connected to the left hemisphere and involved in a variety of cognitive and language functions, including naming, which is often impaired in people with aphasia. The PI's prior work shows that cerebellar tDCS is safe, easily tolerated, and improved language skills in a number of stroke participants with aphasia. The proposed project will build on these findings by conducting a randomized, double-blind, sham-controlled, trial to determine the effectiveness of cathodal tDCS to the right cerebellum for the treatment of post-stroke aphasia. We will compare the effects of 15 sessions of cerebellar tDCS combined with an evidenced-based anomia treatment (semantic feature analysis, SFA) to 15 sessions of sham combined with SFA treatment in patients with chronic (at least 6 months post stroke) aphasia. We will evaluate the effects of cerebellar tDCS on naming untrained items as well as the effects on functional communication, content, efficiency, and word-retrieval of picture description, and quality of life. This project will also identify imaging and linguistic biomarkers to determine the characteristics of stroke patients that benefit from cerebellar tDCS and SFA treatment. Individual response to tDCS treatment is highly variable, and little is known about how factors related to imaging and linguistic characteristics combine to induce treatment responsiveness. We will carry out resting state functional magnetic resonance imaging (rsfMRI), diffusion tensor imaging (DTI), high resolution structural imaging, and detailed linguistic testing before the start of treatment to determine whether these factors can predict response to cerebellar tDCS and/or SFA. The long-term aim of this study is to provide the basis for a Phase III randomized controlled trial of cerebellar tDCS vs sham with concurrent SLT for treatment of chronic aphasia.

NIH Research Projects · FY 2025 · 2022-09

Tools for Gene Editing in Marmosets SUMMARY The goal of this proposal is to create an efficient new set of tools and techniques for the generation of genetically modified marmosets suitable for use as models of brain physiology and function. Development of efficient methods of genetic manipulation in the species will require refinement of genome editing technologies, breeding technologies, ovarian stimulation and oocyte retrieval, embryo culture, and pre-implantation screening of embryos for the desired genetic alternations. A large colony of marmosets will be necessary, along with an infrastructure to refine technologies and an interdisciplinary team with expertise in marmoset care, assisted reproduction technologies, tissue-specific expression with Cre-drivers and CRISPR technologies, and behavior testing and neuropsychiatric analysis. The Specific Aims of this proposal are to: (1) Improve technology for in vitro genetic manipulation of marmosets and explore novel methods for marmoset gene editing; (2) Identify safe harbor loci and establish Cas9, Cre, and GCaMP marmoset lines using promoters for ubiquitous expression, selective expression in brain, or regional or cell-type specific expression; and (3) Generate marmoset models with genetic alterations relevant to the neurobiology of human brain diseases. These studies will provide tools for future gene editing experiments in marmosets, will establish an initial set of genetically modified tool lines for use in the field, and will provide initial proof-of-principle lines for marmoset models of genetic alterations potentially relevant to the neurobiology of human brain diseases.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY Cochlear Hair Cell (HC) loss is a leading cause of noise-induced and age-related hearing loss worldwide. Regeneration of HCs in response to damage has been observed in nonmammalian species, such as birds and fish, but is not observed in the mature mammalian cochlea. Although Supporting Cells (SCs), a diverse population of cells offering metabolic and structural support to hair cells, show the capacity to produce hair cells in neonatal mice, this plasticity is lost by postnatal day 5. Previous work from our lab has identified the de-differentiation of SCs into a more progenitor-like state as a key component of the HC regenerative process, as hair cells and supporting cells arise from a common progenitor pool during cochlear development. Supporting cell de-differentiation, the downregulation in expression of supporting cell-specific genes and upregulation in expression of, allows the progenitor-like cells to respond to HC fate-inducing cues. Preliminary studies have identified members of the NFI and ZBTB families of transcription factors as potential regulators of SC identity, maintaining SCs in a terminally differentiated state and preventing endogenous reprogramming. The NFI factors have previously been studies in the context of retinal regeneration, where they have been shown to regulate retinal Müller glial cell differentiation. Additionally, disruption of NFI factor function has been shown to promote conversion of Müller glia into retinal neurons. Zbtb20 has been implicated in astrocytogenesis, and has been linked to Primrose syndrome, a rare developmental disorder which is known to cause hearing loss. The overarching hypothesis of this grant proposal is that Nfia/b/c/x and Zbtb20 are crucial in maintaining SC identity, and that loss of one or more of these factors will enable SC reprogramming and HC regeneration in stage P5 mouse tissue. This hypothesis will be tested using in vitro cochlear organoid culture derived from stage P2 and P5 murine cochlear sensory epithelia. Additionally, the function of Zbtb20 will be studied in vivo and in a HC damage model. I will use RT-qPCR, fluorescence imaging of a HC reporter line, immunolabeling, EdU pulse experiments, and scRNA-sequencing in Aim 1 to determine if gain of function of NFIA/B/C/X or ZBTB20 inhibits cochlear SC reprogramming and SC-based HC formation at stage P2 in vitro and in Aim 2 to determine if loss of function of NFIA/B/C/X or ZBTB20 enhances cochlear SC plasticity and SC-based HC formation at stage P5 in vitro. In Aim 3, I will use immunolabeling, RT-qPCR, EdU pulse experiments, and a HC damage model to define the role of Zbtb20 in maintaining SCs in a terminally differentiated state. These studies will expand our knowledge of SC identity maintenance and the HC regenerative process and will aid future translational research using regenerative therapies to treat hearing loss in humans.

NIH Research Projects · FY 2024 · 2022-09

Proper development depends on asymmetric cell division (ACD), a process by which dividing stem cells produce a renewed stem cell and a differentiating cell. Many intrinsic and extrinsic factors guiding ACD have been found and characterized. Yet, the contribution of chromatin to cell-fate determination is poorly understood. Previously, our lab discovered asymmetries in histone and histone post-translational modification (hPTM) inheritance in Drosophila male germ cells. Further dissection of this process revealed it functions and is regulated in three-steps: 1) histone asymmetry is established during S-phase; 2) histone asymmetry is distinguished during M-phase; 3) the readout of the inherited asymmetric histones guides cell cycle progression following mitotic exit. Interestingly, disruption of these asymmetries results in both stem cell loss and overproliferation phenotypes, suggesting that asymmetric histone inheritance is an essential process in tissue health. Further, deterioration of this process may be common among diseases including age-related tissue degeneration and cancer. I hypothesize that asymmetric histone inheritance is a general mechanism that plays a crucial role in stem cell homeostasis and cell-fate determination during development. However, our ability to track histones and hPTMs at specific loci within single cells is severely limited at this time. Thus, we are in urgent need of new tools to study the roles and consequences of asymmetric histone inheritance in development, stem cell homeostasis, and cell-fate determination. In this proposal, I will 1) broaden our understanding of pervasiveness, roles, and patterns of histone inheritance in asymmetrically dividing cells and 2) develop a novel method for labeling non-repetitive loci to track epigenomic features. First, I will express histones tagged with the photoconvertible Dendra2 in Drosophila neuronal stem cell lineages. Neuroblasts (NBs), their progenies ganglion mother cells (GMCs), and transit-amplifying intermediate neuronal progenitors (INPs) are a well-studied stem cell model system. Following 405nm light induced photoconversion, I will observe the inheritance patterns of old (red) versus new (green) histones in type I and II NBs. These studies will 1) expand our knowledge on histone inheritance in ACD, and 2) reveal whether histone inheritance patterns change as asymmetrically dividing cells age and lose potency (e.g. INPs). Tracking epigenomic information at specific genomic locations has traditionally been performed using FISH. However, the heat denaturation required for FISH is either partially or completely incompatible with detection of DNA-associated proteins. To overcome this limitation, I will adapt Oligopaint technologies to design guide RNA libraries for tethering dCas9 to specific, non-repetitive loci. This will allow me to track histone and hPTM inheritance patterns at specific genomic loci with single-cell resolution. These works will both enhance our understanding of the mechanisms underlying ACD and provide new tools facilitating the detection of epigenomic alterations in research and clinical settings.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Social isolation is the objective lack of (or limited) social contact with others. Approximately 1 in 4 older adults are socially isolated. Social isolation is a major public health issue and a risk factor for morbidity and mortality comparable to smoking, hypertension, and a sedentary lifestyle. Socially isolated older adults cost Medicare an estimated additional $6.7 billion each year. Low income older adults have two times the odds of experiencing social isolation compared to those with higher incomes. Amidst current aging projections and reports that the current cohort of older Americans have high debt and insufficient savings, the demand for subsidized housing has increased. Few studies examine social isolation among low income older adults living in subsidized housing. Despite epidemiologic evidence demonstrating the deleterious effects of social isolation on health, gaps exist in our knowledge regarding the best solutions to identify and address this major public health problem. This K23 proposal builds upon the candidates’ work funded by the Grants for Early Medical/Surgical Subspecialists’ Transitioning to Aging Research (GEMSSTAR, R03) to advance our understanding of the facilitators and barriers of social isolation among older adults living in subsidized housing. Leveraging these insights, this K23 proposal, guided by the NIH Stage Model for Behavioral Intervention Development, will conduct basic research (NIH Stage 0) to characterize social isolation and its relationship with individual and structural factors among older adults living in subsidized housing to identify participant characteristics that will inform the development of an intervention. These findings will inform the creation (NIH Stage 1A) and piloting (NIH Stage 1B) of a housing-based social isolation intervention pilot. This innovative proposal employs a transdisciplinary approach and builds the foundation for novel approaches to identify and address social isolation among older living in subsidized housing. This proposal will support the research and career development of the candidate, who aims to become an independent clinician investigator focused on developing and implementing community-based interventions that improve the health of low-income older adults.

NIH Research Projects · FY 2024 · 2022-09

Project Summary The goal of this application, submitted in response to PAR-19-368, “Accelerating the Pace of Drug Abuse Research Using Existing Data” is to leverage data from the NIDA Clinical Trials Network to enhance our understanding of treatment effect heterogeneity in psychosocial treatments for substance use disorders. Treatment effect heterogeneity is particularly a concern in research of substance use disorder treatments, in part due to heterogeneous sub-phenotypes of patients in symptom profile, disease course, and recovery trajectory. Nevertheless, analysis of treatment effect heterogeneity in substance use disorder research has been often conducted in a suboptimal manner using subgroup analysis (i.e., estimating impacts separately within groups defined by a single covariate), which could result in finding spurious differences in treatment effects by subgroup due to the performance of multiple statistical tests and random variability across patients. Another challenge of single covariate-based subgroup analysis is that most covariates have small moderating effects and their individual contribution to treatment effect heterogeneity is not meaningfully informative to treatment decisions. Our objective is to apply a novel statistical method, causal forest approach, to systematically examine treatment effect heterogeneity of psychosocial treatments for substance use disorders. This study uses data from 12 randomized controlled trials in the NIDA Clinical Trials Network which examined effectiveness of nine distinct psychosocial treatments against treatment-as-usual condition (Motivational Incentives, Motivational Enhancement Therapy, Screening Motivational Assessment, Therapeutic Education System, Brief Strategic Family Therapy, Twelve-Step Facilitation, Motivational Interviewing, Seeking Safety, and Exercise Program). For each type of psychosocial treatment, we propose to implement the causal forest approach to estimate the expected effect of a treatment for each individual while taking into account multiple covariates simultaneously. The estimated treatment effects will be used to test the presence and degree of treatment effect heterogeneity in each type of psychosocial treatment. Using the variable importance measure obtained from the causal forest, we also plan to identify the most important covariates contributing to treatment effect heterogeneity in each type of psychosocial treatment. These analyses will be repeated for multiple outcomes (e.g., abstinence, reduction in frequency of target substance use) to examine whether and how the degree of treatment effect heterogeneity as well as common effect moderators differ across outcomes. Overall, these analyses will grow the evidence base that can be used by treatment providers to guide treatment decisions for individual patients with substance use disorders.

NIH Research Projects · FY 2025 · 2022-09

Project summary In metazoan, the presence of double-stranded (ds)DNA in the cytosol signals serious problems, which range from radiation damage to pathogen invasion. The innate immune system acts as the first line of defense against cytosolic dsDNA by initiating inflammatory signaling pathways. Cytosolic dsDNA sensing pathways are integral to host defense against numerous pathogens, and their malfunctions also result in various human maladies. Our research program is poised to resolve several long-standing mechanistic questions in understanding how cytosolic dsDNA sensing pathways are activated and regulated at the molecular level. We will also test new concepts as to how these sensors might be stigmatized as autoantigens, and explore the mechanical forces that drive and regulate their higher order assemblies. Our approaches include X-ray crystallography, rigorous biochemical measurements, cell-based assays, electron microscopy, and single molecule methods. Here, we will determine how cGAS coordinates different nucleotide substrates and metal co-factors at the active site to specifically generate 2'-5'/3'-5' linked cyclic G/AMP. We will also investigate how dimerization is allosterically coupled to activation. We will then determine how cGAS and ALR sensors selectively recognize and signal through dsDNA. Next, we will test the role of phase-separation/transition in the normal and aberrant activities of cytosolic dsDNA sensors. Finally, we will delineate mechanical forces and structural mechanisms that underpin the activation and regulation of cytosolic dsDNA sensors. The molecular insights resulting from the proposed studies will provide a foundation for developing new therapeutic strategies that target various human diseases caused by dysregulated cytosolic dsDNA sensing pathways.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY In 2020, an estimated 5.8 million Americans had Alzheimer’s disease (AD), the most common cause of dementia, and this number is expected to reach 13.8 million by 2050. Alzheimer’s Disease and related dementias (ADRD) take an enormous toll on individuals affected, as well as their caregivers. Because disease- modifying therapies have proved largely ineffective, a priority is placed on finding ways to prevent this debilitating disease. Since neighborhood design factors can be modified at the population level, they are an attractive area for intervention. Green space is one neighborhood design element that may be associated with cognitive function because exposure to it may reduce stress, improve mood, promote physical activity, and provide opportunities for social engagement, all of which have been associated with better cognitive health. Some populations are more likely to live in neighborhoods with lower green space than others and design and planning policies have been implicated in the distribution of green space. Recent efforts have increased the amount and quality of green space through park improvement projects, street tree plantings, and vacant lot restoration. However, it is unclear if green space improvements are harmful or beneficial to residents, particularly in neighborhoods that are undergoing change. In the proposed work, we will (1) assess the cross-sectional and longitudinal association between changes in green space and cognitive function, cognitive decline, and dementia over a 20-year period; (2) estimate the association between community planning policies and cognitive function, cognitive decline, and dementia; and determine if this association is mediated by green space exposure; (3) determine if (a) the association between green space and cognitive function, cognitive decline, and dementia is moderated by neighborhood change; and (b) there is an independent association between neighborhood change and psychosocial stressors, mental health, and perceived neighborhood characteristics; and (4) partner with a national network of local non-profit organizations to explore obstacles and facilitators to green space improvements. This work has the potential to provide guidance to improve cognitive health for all Americans.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY / ABSTRACT Stigma is increasingly recognized as a modifiable factor impacting antiretroviral therapy adherence and viral suppression among adolescents and young adults living with HIV (AYALHIV), however few interventions have effectively reduced stigma or address intersectional stigma among this population. In this study we will address internalized HIV stigma among AYALHIV, as well as internalized stigmas related to the prevalent youth experiences of violence and depression. Further, we will also focus on intersectional stigma, capturing anticipated stigma associated with the complex reality of AYALHIV’s multiple social identities. We will create an intervention to reduce these multiple stigmas by building upon the evidence-based intervention Project YES! Youth Engaging for Success. Project YES! is a peer mentoring program that effectively reduced HIV-internalized stigma among 15-to 24-year old AYALHIV in Zambia, and among a subset of pediatric clinic participants, increased their viral suppression. Project YES! data also highlighted critical intervention gaps, including high levels of physical, sexual and/or emotional violence victimization among AYALHIV, and how participants who screened positive for depression had lower levels of viral suppression. This data aligns with a growing body of evidence linking violence victimization and depression with internalized stigma and viral failure. Project YES! peer mentors would refer participants with violence experiences or depression and anxiety to the clinic health staff in a system that provides minimal mental health training or services for AYALHIV. This research seeks to address this gap by integrating the stigma reducing EBI youth peer mentoring program (Project YES!) with the World Health Organization endorsed and proven mental health approaches that reduce symptoms of depression and anxiety (Problem Management Plus (PM+) and Early Adolescent Skills for Emotions (EASE)) that are delivered by lay health workers. By adapting and integrating PM+/EASE to create Project YES+, we will specifically provide AYALHIV with problem-management skills that we hypothesize will interrupt the bi-directional pathways among violence victimization, depression and internalized and intersectional stigmas. We will also refine our measures of internalized stigmas related to violence and depression, and intersectional stigma. Specific study aims include: AIM 1: Adapt and integrate the WHO problem management-based PM+/EASE approaches for AYA (15-24 years) to create Project YES+ for implementation by lay youth mentors to improve viral status by reducing the internalized and intersectional stigmas related to HIV, violence, and depression. Aim 2: Use cognitive interviewing to adapt scales of violence- and depression-related internalized stigma and intersectional stigma to improve stigma measurement among AYAHLIHV. Aim 3: Assess the feasibility and acceptability of a Project YES+ intersectional stigma reduction intervention among 100 AYALHIV through a pilot randomized controlled trial.

NIH Research Projects · FY 2024 · 2022-09

ABSTRACT The primary goal of this project is to further our understanding of the contribution of the vestibular system to balance control, particularly the neural mechanisms driving reflexive responses to postural perturbations. Visual, proprioceptive, and vestibular signals are the primary means for sensing the position of the body and maintaining balance. However, the central neural mechanisms of postural control remain elusive, as such research requires recording single neurons in the brainstem during free behavior. In order to understand the vestibular component of central postural control, I will first establish baseline postural responses in an animal model, the rhesus macaque, which has proven to be a valuable model for understanding the neural control of human vestibular function and processing. I will then characterize behavioral and neural responses during these posture-stabilizing behaviors in healthy animals, bilateral vestibular loss animals, and vestibular loss animals with replacement-of-function using a vestibular prosthesis. Aim 1 of this project is to characterize behavioral responses to support surface perturbations. For postural perturbation experiments, the animal is acclimated to a behavioral chamber mounted on a 6-degree-of-freedom hexapod motion platform. The hexapod delivers support surface perturbations, and the animal’s motion is tracked using inertial measurement units, a force plate, and markerless video motion tracking. These perturbations are repeated in normal and bilateral vestibular loss animals, and the animals’ postural responses are quantified in order to build a model of the vestibular contribution to balance control. Aim 2 is to characterize responses of vestibular-sensitive neurons that drive postural reflexes to support surface perturbations. I will leverage emerging technology in wireless, high-density neural recording to characterize the responses of neurons in the vestibular nuclei (VN) to postural perturbations in order to elucidate the contribution of vestibulo-spinal reflexes, which are driven by VN cells, to postural corrections. Finally, for Aim 3 I will repeat these experiments and recordings with an animal fitted with a multichannel vestibular prosthesis in order to assess improvements in posture caused by the vestibular prosthesis, as well as neural adaptation to novel and/or modified vestibular inputs delivered by the prosthesis. This work has the potential to contribute to our understanding of the role of vestibuloception in maintaining upright posture, as well as improve quality-of-life for patients experiencing bilateral vestibular loss by providing data on the best modulation strategies for vestibular prostheses.