Research Inst Nationwide Children'S Hosp

NIH Research Projects · FY 2026 · 2024-06

Abstract Approximately 30% of hepatitis C virus-infected persons spontaneously clear the virus within six months. The remaining 70% develop chronic infection. Of those with chronic HCV infection, the risk of cirrhosis ranges from 15% to 30% within 20 years. The viral and immune correlates of these different outcomes of infection and pathogenesis remain poorly studied, mainly due to the lack of an informative animal model. We developed surrogate animal models for HCV using a rodent HCV-like virus isolated from feral brown rats (Rattus norvegicus), RHV-rn1 (Rn-1). So far, studies of Rn-1 infection and immunity in lab mice have revealed that normal immunocompetent mice do not develop prolonged or chronic HCV-like infections. Notably, chronic HCV and RHV infections can be studied in their natural host, humans and rats, respectively. However, how HCV evades immunity during the acute phase, a prerequisite for developing chronic infection, requires animal models that allow comparative analyses of HCV-like clearance and persistence and mechanistic analysis of innate and adaptive immune responses. To create such models, we isolated several new RHV variants from feral rats and used these as a pool for serial passaging and adaptation in mice. After several passages, we identified a mouse- adapted variant (Rn-2m) that produced delayed clearance or chronic infection in normal lab mice. Subsequently, we identified the parent rat virus Rn-2, and its infection cleared within 3-4 weeks in mice. The project seeks to identify the viral and immune determinants of heterogenous outcomes of HCV infection and pathogenesis. Considering the strengths and weaknesses of the rat and mouse models, we propose a comparative biology approach where the new Rn-2m infection in the mouse model, due to the availability of vast resources, will be used to develop and test hypotheses, and the rat model will be used for validation of key findings. Specific Aim 1 is to identify the viral correlates of spontaneous clearance and chronic infection. Our new data indicate that Rn-2m constantly evolves and modulates innate immune cells in the liver of infected mice to establish a prolonged or chronic infection. We will compare the nature of infection, evolution, and immunogenicity of clone- rescued Rn-2 and Rn-2m to identify viral correlates of acute clearance, delayed clearance, and chronic infection in mice and subsequently in rats. Specific Aim 2 is to identify the immune correlates of chronic infection and liver diseases. We will use RHV-specific mouse and rat MHC tetramers and E2 protein tetramers, ex vivo antigen stimulation, passive antibody, and in vivo T and B cell transfer, and cell depletion experiments to define the role of T and B cells in determining different outcomes of RHV infection and pathogenesis. We expect that the solid rationale behind our approach will yield a biologically relevant and widely accessible lab mouse model for HCV and gain novel and mechanistic insights into HCV immune evasion and pathogenesis. This knowledge is crucial for developing new strategies to prevent chronic HCV infections and associated liver diseases.

NIH Research Projects · FY 2026 · 2024-02

Abstract Metastsis is the direct cause of the majority of deaths in patients with solid tumors, thus representing a major unmet medical need. In some cancer types, microscopic metastases may have already occurred at the time of diagnosis. Because an established tumor microenvironment mediates therapeutic resistance through a variety of mechanisms, cancer cells are likely most vulnerable during their time in the bloodstream and early in the development of micrometastasis. Thus, effectively targeting those cells requires constant surveillance in order to “catch” them either in the circulation or in their early metastatic niche, before they have re-established a protective microenvironment. Currently FDA-approved systemic therapies for solid tumors are unable to adequate do so as they are intermittent; chemotherapy is given in cycles to allow recovery from toxicity to bone marrow and other organs, and even long-half-life antibodies result in peaks and troughs. Collectively, our available therapies likely fail to provide systemic tumor control of metastases in part because intermittent exposure not only kills only a percentage of cells during each treatment cycle, enabling surviving cells to develop cell-intrinsic drug resistance, but also due to the drug-free windows that enable cancer cells to spread. Our overarching hypothesis is that we can prevent and treat micrometastasis by leveraging gene therapy to provide constant, long-term, systemic immunologic pressure on cancer cells. We will test our hypothesis in models of neuroblastoma, the most common solid tumor in children outside of the brain, and Her2+ breast cancer, both of which have a poor prognosis when metastatic. We have developed an off-the-shelf strategy utilizing a single intravenous dose of recombinant adeno-associated virus (rAAV) to instruct normal cells to secrete bispecific T cell engagers into the bloodstream for long periods of time (rAAVrh74-aGD2-aCD3 and rAAVrh74-aHer2- aCD3), a platform we term “TransJoin.” Our specific aims are to: (1) Test the anti-metastasis efficacy of GD2 and Her2 TransJoins in preclinical models, (2) Test the safety of GD2 and Her2 TransJoins in preclinical models, and (3) Develop controllable expression of GD2 and Her2 and a kill switch. If successful, our data will support clinical translation of rAAVrh74-aGD2-aCD3 for neuroblastoma and rAAVrh74-aHer2-aCD3 for Her2+ breast cancer. Following a human phase I safety study, we envision clinical trials during post-induction therapy in the setting of minimal residual disease, to either prevent and/or treat micrometastases and improve long-term patient survival.

NIH Research Projects · FY 2026 · 2024-02

Congenital heart defects (CHD) are the most common type of birth defect, with cardiac malformation resulting from abnormal heat development contributed by genetic and/or environmental risk factors. The association of maternal diabetes with increased offspring CHD incidence is proposed to be attributed to factors including glucose imbalance and metabolic dysregulation. Murine studies on hyperglycemia have been beneficial in uncovering the phenotypic effects of maternal hyperglycemia on embryonic development, with reports that high glucose conditions are associated with general suppression of WNT signaling, apoptosis, and excessive reactive oxygen species (ROS) production. However, little is known about the effects of hyperglycemia on human stem cells in regard to cardiomyocyte determination, metabolism, and functionality. Additionally, there is little reports of these alterations using induced pluripotent stemcells (iPSCs), which provide an opportunityto observe cardiac progenitor cell (CPC) heterogeneity, multicellular crosstalk, and differentiated cell functionality at key development timepoints. We hypothesize that high glucose environments interfere with normal cardiac differentiation to cause altered cell lineage determination in iPSCs and iPSC-derived cardiomyocytes (iPSC- CMs). We propose use of immunofluorescence assays and staining to quantify mitochondrial metabolism, mitochondrial ROS production, and apoptosis marker amounts. Our goal is to investigate the impact of high glucose on human cardiomyocyte differentiation and crosstalk signaling abnormalities during 3D stem cell differentiation that could lead to impaired cardiac development. We will investigate differentially expressed genes at the single-cell resolutionin early and late differentiation in responseto high glucose doses in iPSC populations. Should our transcriptional profiling comparisons yield iPSC and iPSC-CM expression differences, the resulting phenotypes would provide insight into cardiac cell determination during hyperglycemic conditions. Understanding the hyperglycemia-response mechanisms is important in the onset of maternal hyperglycemia- associated CHD. These mechanisms would be further investigated through looking at cell lineage population composition in organoids and analyze response per lineage to narrow down what structures of the mature heart are affected.

NIH Research Projects · FY 2026 · 2024-02

Project Summary/Abstract Variants in the tubulin folding cofactor D (TBCD) gene result in a rare early-onset encephalopathy with neurodevelopmental and neurodegenerative features including developmental regression, epilepsy, microcephaly, hypotonia, and spasticity which progresses to immobilization, ventilation, and premature death. TBCD functions as a tubulin-specific chaperone, which plays a crucial role in regulating microtubule dynamics throughout neurodevelopment. Since first reported 7 years ago, modest progress has been made in understanding molecular and cellular mechanisms contributing to disease pathogenesis and therapeutic development. Our team has recently established and characterized the largest known cohort of induced pluripotent stem cell (iPSC) lines from a phenotypic spectrum of 5 TBCD patients, from whom we gathered corroborative clinical data. We show TBCD patient-derived cerebral organoids (COs) display a phenotypic spectrum of profound growth deficits, microtubule instability, and early neurodegeneration, highlighting the value of COs to recapitulate TBCD patient phenotypes. To compare the effect of different pathogenic variants on clinical, molecular, and biochemical disease progression in vivo, we have generated a novel allelic series of TBCD variant knock-in mouse models. We propose the central hypothesis that TBCD deficiency causes microtubule instability and cell-type specific dysfunction during critical milestones of neurodevelopment, which include molecular deficits in progenitor proliferation, migration, neuronal differentiation, and ultimately neurodegeneration. Through the use of novel transgenic mouse and complementary organoid models, we will test this hypothesis and in parallel establish a strong framework for developing a gene replacement therapy for patients with TBCD deficiency. In Aim 1, we will determine neuropathological mechanisms of TBCD variants using patient derived COs including through use of electrophysiology, single cell RNAseq, and CRISPR gene correction. In Aim 2, we will comprehensively investigate the molecular and clinical phenotypes of a novel allelic series of TBCD variant knock-in mouse models utilizing clinically relevant outcome measures such as CatWalk XT® gait analysis, MRI, and EEG. In Aim 3, we will determine the therapeutic efficacy of targeted AAV-mediated gene replacement by modulating the cell specificity, expression level, and neurodevelopmental stage in which functional TBCD is delivered. To enhance translation and validate that the lead construct provides comparable TBCD expression in the context of human brain cell types, we will treat previously established patient-derived COs with the lead AAV9-TBCD construct and assess for phenotypic rescue. Utilizing this rigorous and multidisciplinary approach, we aim to uncover molecular and cellular mechanisms underlying the profound deficits in TBCD-related developmental and epileptic encephalopathy; establish the first tractable in vitro and in vivo models of TBCD; and evaluate a highly translatable and potentially transformative AAV-mediated gene replacement therapy.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT Congenital cytomegalovirus (cCMV) infection is the most common congenital viral infection in the U.S., affecting approximately 18,500 liveborn infants annually. Children with symptomatic or asymptomatic infection at birth can suffer from permanent neurodevelopmental disabilities including cerebral palsy, intellectual impairments, visual deficits, and sensorineural hearing loss. Treatment with the antiviral drug, valganciclovir, improves neurologic outcomes, but drug adverse effects preclude universal antiviral treatment for all cCMV infected infants. Among adults with primary CMV infection, natural killer (NK) cells serve as the first antiviral defense and are rapidly followed by T cell responses that effectively control viral replication. In contrast, cCMV infected infants lack robust antiviral T cell responses to cCMV infection over the first year after birth. These T cells express an exhausted phenotype and fail to control viral replication for months to years. In-utero NK cell responses arise during cCMV infection, but the contribution of these cells to viral control and neurologic outcomes are unstudied. We compared the NK cell immunophenotypes of cCMV infected and uninfected neonates by CITE-seq and found 4 clusters of cytotoxic NK cells induced by cCMV infection. These clusters expressed both activating and inhibitory receptors associated with both NK cells and T cells, and one cluster expressed an inhibitory signature typically observed during T cell exhaustion. Flow cytometry confirmed that cCMV-induced NK cells expressed inhibitory receptors, but NK cells of CMV seropositive adults did not. We therefore hypothesize that in-utero CMV infection induces novel cytotoxic NK cell responses with adaptive-like features that differ from adult responses. We further propose that cytotoxic NK cell function differs between cCMV infected individuals, is altered by antiviral treatment, and may correlate with neurologic outcome. In this project we will characterize the adaptive-like features of cCMV- induced NK cells using blood samples collected longitudinally from a large cohort of cCMV infected infants, CMV uninfected infants, and CMV seropositive adults. Aim 1 will define the immunophenotype and cytotoxic potential of NK cells induced by cCMV infection using CITE-seq, spectral flow cytometry, and in-vitro cytotoxicity assays. Aim 2 will determine the effect of antiviral treatment upon cytotoxic NK cell quantities and immunophenotypes and explore potential associations between NK cell parameters and cCMV neurologic outcomes. Together, these studies will elucidate novel features of NK cell responses induced by in-utero viral infection that differ from adult responses and investigate their relationship to clinical treatments and outcomes among children with congenital CMV infection.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY AND ABSTRACT: Chronic musculoskeletal pain (CMSKP) is common in adolescents. Participation in regular physical activity (PA) is effective in reducing CMSKP but PA participation is still below recommended levels. There is a need to better understand ways to better promote PA in adolescent CMSKP (e.g., decreasing sedentary activity vs. increasing moderate to vigorous PA), factors that affect PA (e.g., resilience, pain modulation, stress dysregulation), and the mechanistic underpinnings of how these factors affect PA and pain. This knowledge may lead to the development of improved interventions for CMSKP. Psychological resilience factors that build on positive attributes, such as self-efficacy, motivation, and psychological flexibility may help adolescents to engage in PA. While resilience factors are important for overall pain coping, they only recently are being considered as predictors for PA. The overall goal of this proposal is to develop and pilot a feasibility study of an intervention that increases resilience for PA in adolescents with CMSKP. This is significant because the development of such an intervention may lead to improved PA and subsequently improved chronic pain for adolescents with CMSKP. This study first evaluates pathways of resilience factors on PA and pain in adolescents with CMSKP (Aim 1), and the modulation of pain and stress. 60 adolescents with CMSKP will complete self-report measures of resilience and vulnerability factors, and objectively measured PA, pain (Quantitative Sensory Testing), and stress (Cortisol Awaking Response). We will evaluate whether endogenous pain modulation and stress regulation mediate the relationships between resilience and PA. This study also will consider resilience factors that are significant in predicting less sedentary PA as intervention targets, and it will develop, refine (Aim 2) and (Aim 3) conduct a pilot feasibility and acceptability study of a resilience-focused chronic pain intervention for youth (Pain REsilience Promotion for Youth [PREP-Y]). The manual will be developed based on an established treatment program for adolescent CMSKP in adolescents (i.e., FIT Teens) and modified to target resilience mechanisms and sedentary activity. This will be done with integrated feedback from content experts in pediatric pain and CMSKP patients. The intervention will then be provided to 30 patients in groups of 4 to 6. This project will be implemented as part of a strong career development plan consisting coursework, experiential training, and intensive mentorship from a team of experts in pediatric behavioral intervention development and evaluation, psychological factors of pediatric pain, physical activity intervention, and pain physiology and assessment. Completion of this study will occur across two academic medical centers with a joint department of pediatrics, a well-establish pain treatment service, and research institutes to promote the growth and development of research faculty. This study and career development plan will promote the establishment of independent, extramurally funded clinical-research program focused on improving clinically meaningful outcomes for adolescents CMSKP.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Advances in childhood cancer therapies have resulted in >80% five-year survival rates.1 With nearly 500,000 childhood cancer survivors in the U.S., there is a growing need to address and mitigate late effects of treatment.2,3 Fertility impairment occurs in about half of male childhood cancer survivors and can negatively impact quality of life.4,5 Sperm banking is an established and widely available form of fertility preservation.6 Despite recognizing the value of future parenthood and guidelines to bank sperm before treatment,6,7 banking remains underutilized among male adolescents with cancer, especially at hospitals with low resources.8-10 Studies show adolescent males are more likely to bank sperm prior to cancer treatment if: 1) their institution has dedicated fertility services,11 2) their provider recommends banking,12 and 3) their parents recommend it.13 To improve equitable care and access to sperm banking opportunities,10 family-centered decision tools, and strategies to implement these tools, are critical. Our team developed the Family-centered Adolescent Sperm banking Tool (FAST) and tested it prospectively among families of adolescent males newly diagnosed with cancer. We found the FAST prompted deeper thinking and facilitated family communication about parenthood goals and banking,17 and that sperm banking rates increased significantly.19 Given feasibility and equity considerations (i.e., short time frame, variety of settings in which fertility counseling is delivered), the objective of this proposal is to adapt the FAST to a web-based tool that can be accessed in inpatient/outpatient/non- clinical settings, identify implementation strategies for high and low-resourced settings, and test these in a three-site pilot study among 110 families of adolescent males (13-21 years of age) newly diagnosed with cancer. Specific aims are to: 1) adapt the FAST (design and delivery) based on patient and caregiver stakeholder feedback (usability testing), which will result in a web-based tool that is functional, acceptable to families, and feasible to implement pre-cancer treatment; 2) conduct rapid contextual inquiry using a mixed- methods approach with clinician stakeholders, to identify tailored implementation strategies for the web-based FAST in both high and low-resourced pediatric cancer centers, prepare for adoption into clinical pathways, and ensure fidelity and sustainability, and 3) pilot the web-based FAST and tailored implementation strategies locally and at 2 additional sites (1 high and 1 low-resourced) to examine feasibility, acceptability, and impact on banking rates (pre-/post design) to prepare for a larger multi-site hybrid implementation-effectiveness trial. Findings will result in an innovative decision tool that can be easily accessed on the web and implemented at a wide variety of high and low-resourced settings to facilitate banking decisions among adolescent males with cancer. These efforts will optimize long-term outcomes by expanding opportunities for parenthood and contributing to improved quality of life in a growing population of survivors, including underserved populations.

NIH Research Projects · FY 2025 · 2023-09

Acute seizures may evolve into status epilepticus (SE), i.e., prolonged or repeated seizures without regaining consciousness, resulting in irreversible brain injury or death costing ~$4 billion annually to the US healthcare system. If seizures are treated rapidly with a correctly-dosed benzodiazepine (BZD), SE and its associated morbidity and mortality may be averted. For instance, children treated after 10 minutes of seizure onset are 11 times more likely to die during their hospitalization than patients treated earlier. Therefore, the American Academy of Neurology identified time to treatment as a quality metric to evaluate SE care. Despite these evidence-based recommendations, delayed seizure treatment remains the status quo in many centers. The Quality Improvement in Time to Treatment of Status Epilepticus (QuITT-SE) study will examine the implementation and effects of a standardized set of QI interventions across pediatric hospitals with diversity in geography, patient demographics and resources. The proposed interventions were developed and tested in a single-center study that doubled the number of SE episodes treated within guideline timing, decreased morbidity, and mitigated over $2 million of charges in the following 17-month period. Our overarching goal is to study the implementation of these QI interventions and their effect on time to SE treatment across multiple centers utilizing an effectiveness-implementation hybrid design. The QuITT-SE trial will be a stepped-wedge cluster randomized trial across multiple sites within the Pediatric Status Epilepticus Research Group, an established consortium of hospitals with expertise in studying and treating SE in children. During this project, mixed qualitative and quantitative methods will be used to identify implementation factors related to the interventions. In addition, the use of standardized QI processes within the Practical, Robust Implementation and Sustainability Model (PRISM) will facilitate the identification of site-specific drivers and themes pertaining to delayed SE treatment, improving the generalizability of findings. The specific aims are: 1) Primary: Determine how implementing a QI bundle impacts the time to treat SE among hospitalized, non-critically ill children. 2a) Secondary: Determine the effect on Pediatric Cerebral Performance Category score among hospitalized, non-critically ill children after implementing a QI bundle. 2b) Secondary: Determine the effect of dissemination of a QI bundle on cost of hospitalization for SE among hospitalized, non-critically ill children 3) Exploratory: Explore the factors implicated in implementing a QI bundle on the time to treat SE among hospitalized children. Public Health Impact: Each year, SE results in an estimated 15,000-45,000 deaths in children in the US and costs ~$4 billion, increasing markedly with more prolonged seizures. Successful completion of QuITT-SE will produce a set of evidence-based interventions alongside an implementation framework to improve outcomes and value of SE treatment across diverse hospital settings.

NIH Research Projects · FY 2025 · 2023-09

This proposal’s overall goal is to hasten drug development for children < 5 years with congenital muscular dystrophy secondary to laminin α2-related dystrophies (LAMA2-RD) mutations. Excellent mouse models of differing severity improved the understanding of pathogenesis in LAMA2-RD. Therapeutic strategies, including protein replacement and apoptosis inhibition (Phase 1), linker gene transfer, and compensatory gene upregulation (pre- clinical proof of concept), are all at various developmental stages but are expected to come to clinical trials in 2-3 years. While all these advances are promising, currently, no validated clinical outcome assessments (COA) are available for children with LAMA2-RD < 5 years. Thus, the need to validate outcome measures and biomarkers is urgent for children (< 5 years) with genetically confirmed LAMA2-RD. Successfully translating any therapy must include these youngest children for whom strength or function-based approaches designed for older “cooperative children” do not work. Clinical trial readiness for infants and young children is particularly critical since therapeutic interventions, if successful, are likely to have the best response when given early. The specific aims of our proposal are to 1) Validate motor function as COA for children with LAMA2-RD, 2) Establish minimal clinically important differences for motor COAs by anchoring them to the clinical global impressions scale, 3) Determine what cohort characteristics will best inform clinical trial eligibility, and 4) Validate novel biomarkers (cross-sectionally measure biceps and rectos femoris by ultrasound) and creatine kinase levels over time. To achieve these aims, we propose a 14-site multicenter prospective 2-year study of 44 children < 5 years at enrollment. Detailed training of at least two clinical evaluators from each collaborating site will take place at the lead institution, Nationwide Children’s Hospital, before enrollment and again in Year 3. We selected the sites based on their expertise in pediatric neuromuscular clinical trials. LAMA2-RD is ultra-rare, and these children are often medically fragile. Therefore, we also selected geographically diverse locations to minimize travel and burden of trial participation. A novel COA developed by necessity during the COVID-19 Pandemic is video assessments of all motor function COAs, further allowing less travel for children. Our partnerships with advocacy groups, including Cure CMD (Congenital Muscular Dystrophy) and the Muscular Dystrophy Association, will allow us to successfully recruit children using a spoke and hub model. The proposal will develop and validate COAs for children < 5 years with LAMA2-RD and will inform future clinical trial design and interpretation. Furthermore, once validated, these COAs are very likely to be successful for children with other rare disorders affecting motor development in early infancy.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT The prevalence of symptom-based diagnosis of gastroesophageal reflux disease (GERD), its empiric treatment, and practice variation in high-risk hospitalized infants remain high. Prior clinical trials data lack physiological- based, objective diagnostic evidence and treatment. GERD is frequently diagnosed by inadequate criteria, and the relative risks, benefits, and indications of GERD therapies are unclear. Thus, gaps in knowledge persist, and a GERD diagnosis can cost an extra $70k and 30 days of hospitalization per infant. Objective, effective, evidence-based therapies are critically needed. As nearly all infants in the neonatal intensive care unit (NICU) are symptomatic, twenty-four-hour pH-Impedance (pH-Imp) testing can diagnose objectively and detect mechanisms of pathophysiologic GERD. Therefore, this study’s overall objective is to conduct a single-center randomized control trial (RCT) to test the central hypothesis that the effects of common therapies [natural maturation, acid suppression-proton pump inhibitor (PPI), or thickened feeds using added rice (AR) formula] are distinct in high-risk infants, under six months corrected maturational age with pH-Imp defined criteria for GERD. The rationale is to refine novel diagnostic criteria and rigorously investigate the impact of randomized therapies to achieve the long-term goal of creating effective, simplified, evidence-based, and scalable treatment algorithms. We will test this hypothesis in the following Specific Aims: Aim 1: Conduct the RCT to evaluate and compare the efficacy and safety of interventions on the primary clinical endpoint (oral feeding improvement and absence of troublesome symptoms) to test the working hypothesis that AR is superior to natural maturation and PPI. Aim 2: Evaluate pH-Imp characteristics and test the working hypotheses that pH-Imp mechanisms are distinct between a) assigned therapies, and b) clinical success and failure. This project challenges current empiric approaches to diagnosing and managing GERD in NICU infants by using both a novel combination of pH-Imp metrics and randomized allocations with targeted stratification (GERD severity and feeding skills) to determine true therapeutic effects on clinical and mechanistic characteristics. The anticipated outcomes will identify diagnostic algorithms for determining GERD and effective treatment approaches. In addition, the proposed research is significant because the knowledge gained will provide the diagnostic basis, framework, and scientific and economic rationale for future studies to further refine therapies across the pediatric age spectrum. We expect new benchmarks to develop preventative and therapeutic strategies for GERD, which will positively impact the quality of life for infants and parents by improving digestive health, overall nutrition, and growth, all relevant to the mission of NIH (NIDDK).

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY / ABSTRACT The goal of the ECHO Ohio Cohort Site at Nationwide Children’s Hospital and The Ohio State University Wexner Medical Center (NCH-OSUWMC) is to contribute to the ECHO Cohort by collecting and providing valuable data on a diverse and often underserved population of participating pregnant individuals, conceiving partners, and children who live in our region, so we can improve maternal and child health everywhere by better understanding how exposures prior to and during pregnancy impact childhood outcomes. Nearly 1/5 of Ohio’s children live in poverty. The rate is more than twice that for non-Hispanic Black children. Central Ohio is diverse with a rapidly growing immigrant population including the largest ethnic Nepali and 2nd largest Somali refugee populations in the U.S.; 1 in 6 Columbus children have an immigrant parent. The patients we serve live not only in urban, suburban, and rural central Ohio, but also within Appalachian portions of southeast Ohio, Pennsylvania, and West Virginia. The communities we serve are ranked poorly across multiple maternal and child health well-being indicators, underscoring a critical need to better understand environmental influences in the perinatal period that contribute to adverse child outcomes both locally and nationally. The NCH-OSUWMC health system is highly experienced in contributing to large multicenter NIH studies involving pregnant individuals and children and well- poised to contribute interdisciplinary leadership. We have maintained a large perinatal repository of clinical, survey, and rich biospecimen data from pregnant individuals, their partners, and their children for over a decade. Our research coordinators and investigators are highly experienced in recruiting both pregnant individuals and children for research investigations and securely and efficiently processing health data and biospecimens. Of note, our track record in retaining diverse subpopulations of high-risk maternal/child dyads, as well as conceiving partners, for follow-up is strong. We provide specific expertise in evaluating lifestyle exposures in pregnancy, most prominently in maternal cardiovascular health, and outcomes expertise in pre-, peri-, and postnatal health and childhood neurodevelopment. We propose 1) evaluating the impact of maternal cardiovascular health during pregnancy using the American Heart Association Life’s Essential 8 framework on child socioemotional development and behavior to age 21, using existing ECHO Cohort Protocol core data elements and 2) investigating, using innovative methods (continuous glucose monitoring), the association between evolving maternal dysglycemia patterns across the peripartum period and child socioemotional development and behavior while evaluating neonatal anthropometrics as potential mediators. We propose to evaluate the interaction between genes and lifestyle exposures on socioemotional development via an association study complemented with imputed -omics data. In a preconception-focused aim, we propose examining the impact of maternal and paternal preconception cardiovascular health on socioemotional development and behavior. Our contributions to the ECHO cohort, providing diversity and expertise, will enhance knowledge leading to improved child health.

NIH Research Projects · FY 2025 · 2023-09

The goal of the ECHO Ohio Cohort Site at Nationwide Children's HospItal and The Ohio State University Wexner Medical Center (NCH-OSUWMC) is to contribute to the ECHO Cohort by collecting and providing valuable data on a population of participating pregnant participants, conceiving partners, and children who live in our region, so we can improve maternal and child health everywhere by better understanding how exposures prior to and during pregnancy impact childhood outcomes. The NCH-0SUWMC health system is highly experienced in contributing to large multicenter NIH studies involving pregnant and child participants and well-poised to contribute interdisciplinary leadership. We have maintained a large perinatal repository of clinical, survey, and rich biospecimen data from pregnant participants, their partners, and their children for over a decade. Our research coordinators and investigators are highly experienced in recruiting both pregnant participants and children for research investigations and securely and efficiently processing health data and biospecimens. Of note, our track record in retaining maternal/child dyads, as well as conceiving partners, for follow-up is strong. We provide specific expertise in evaluating lifestyle exposures in pregnancy, most prominently in maternal cardiovascular health, and outcomes expertise in pre-, peri-, and postnatal health and childhood neurodevelopment. We propose 1) evaluating the impact of maternal cardiovascular health during pregnancy using the American Heart Association Life's Essential 8 framework on child development and behavior to age 21, using existing ECHO Cohort.Protocol core data elements and 2) investigating, using innovative methods, the association between evolving maternal dysglycemia patterns across the peripartum period and child socioemotional development and behavior while evaluating neonatal anthropometrics as potential mediators. We propose to evaluate the interaction between genes and lifestyle exposures on development via an association study complemented with imputed-omics data. In a preconception-focused aim, we propose examining the impact of maternal and paternal preconception cardiovascular health on development and behavior. Our contributions to the ECHO cohort will enhance knowledge leading to improved child health.

NIH Research Projects · FY 2025 · 2023-09

Summary The Midwest Pediatric Device Consortium (MPDC) has been created to address unmet needs in pediatric medical device development. Founded at Nationwide Children’s Hospital (NCH) and The Ohio State University (OSU) in Columbus, Ohio, MPDC will use funds from the FDA Pediatric Device Consortia (PDC) program in support of its mission to provide funding and business, legal, organizational, clinical, engineering and scientific resources to inventors of pediatric medical devices having significant commercialization potential and that address pediatric health needs. NCH and OSU both maintain national prominence in clinical care and research. Teaming with other pediatric healthcare organizations including Cincinnati Children’s and Cleveland Clinic Children’s establishes a vibrant and comprehensive medical device development ecosystem for our region and provides diversity in leadership, participation, community engagement and economic opportunity. MPDC is also powered by Ohio’s Trade Organizations, including Jobs Ohio and Ohio Life Sciences, to leverage their network of connections and state legislative influence. Lastly, MPDC engages with several venture funds including Rev1 Ventures, CincyTech, and Project MedTech, having considerable resources and experience bringing startup companies forward. Participating organizations of the MPDC have already assisted a large number of devices along the development pathway, demonstrating the business experience and stewardship of ideas necessary for programmatic success. The full range of support is offered to innovators by the MPDC network, comprised of preeminent experts in their field who will provide advice, education, guidance and funding to help lead device development through the pipeline to commercial success. Our Executive Committee and Advisory Committee are made up of clinicians, scientists, business and manufacturing experts who are thought-leaders in their fields. They will evaluate the clinical and business merits of every device project submitted for evaluation and funding. MPDC’s organizational structure includes a broad spectrum of leaders who are able to assess the value proposition, reimbursement, regulatory, marketing and business strategy of each proposed device as a cost-free service. MPDC will implement its comprehensive plan to foster collaboration, diversity, equity, and inclusion as a leader in creating a national network benefiting all pediatric patients MPDC will also provide unique real world evidence through projects specifically crafted to capture data streams as well as develop mechanisms to collect new data to support pediatric device label expansion. MPDC will be a beacon of pediatric medical device innovation through its bridging of academic researchers, inventors, entrepreneurs and industry partners in an integrative program to transform pediatric healthcare through advancement of new and needed technologies.

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract Burn injuries continue to be one of the leading causes of unintentional death and injury in the United States. Non-fatal burn injuries are a leading cause of morbidity with the most common complications being infectious related. Thermal injures result in massive fluctuations in the inflammatory and immune responses (e.g., plasma cytokines, cellular phenotypes, immune function, and soluble proteins) and these changes have been associated with mortality and infectious complications. However, the underlying mechanisms driving these dysfunctional responses is still largely unknown both systemically and locally. Our primary goal is to investigate the systemic and localized mechanisms of immune dysfunction following pediatric thermal injury to provide useful biomarkers to identify those at highest risk to develop subsequent infection with a goal to develop therapies to improve patient outcomes. Over the next five years we will pursue three integrated research plans that use both pediatric blood and tissue samples as well as an established juvenile murine model of burn injury and infection, to investigate the mechanisms of immune dysfunction after burn injury. Research Plan 1 will investigate the mechanisms underlying lymphocyte dysfunction after severe burn injury. In this work we will assess the cell surface expression of lymphocyte co-inhibitory molecules, examine apoptotic markers on isolated lymphocytes, and perform functional assays. This work will provide insight into the mechanisms surrounding lymphocyte dysfunction and guide future immunomodulatory therapies that can target these specific mechanisms. Research Plan 2 will evaluate the differentiation of systemic monocytes to tissue macrophages after severe burn injury and post-burn complications. In this work we will perform functional assays on isolated systemic monocyte populations and isolated tissue macrophages. This work will lead us to a more complete mechanistic understanding of the differentiation of systemic monocyte populations to the localized macrophages in the wound bed. This information will inform the best route of administration (systemic or topical) to apply immunomodulatory therapy to augment these macrophages and improve tissue level outcomes. Research Plan 3 will elucidate the potential benefits of FDA-approved immunomodulator therapies after severe burn injury to prevent post-burn complications. This work will use combination therapies, ex vivo in pediatric samples and in vivo in our murine burn model, to assess changes in immune function. The findings from this study will be important to advance the design of future immunostimulatory clinical trials in this population as well as other forms of surgical predisposition to sepsis such as trauma and major surgery. Moreover, this could serve as a model for other age groups of burn injuries. Together, these projects will provide an important building block for developing appropriate therapeutics in treating burn wounds that can simultaneously help control inflammation, prevent infection, and assist in wound healing, which is an absolute necessity for burn care research.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY/ABSTRACT: Congenital heart defects (CHD) affect over 2.4 million individuals in the U.S and generate $6.1 billion in annual healthcare costs. Avoidance of tobacco is vital to mitigate CHD survivors’ risk for acquired cardiovascular complications, but the youth e-cigarette (“vaping”) crisis exposes the growing and vulnerable population of adolescents (“teens”) with CHD to potential vaping-related harms and significantly increases risk for conventional cigarette use. Interventions to prevent vaping among teens with CHD are critically needed, but no existing programs are sufficient for this at-risk group. Dr. Fox’s TL1 fellowship research showed that 31% of teens with CHD are susceptible to vaping and identified perceived stress and poor disease knowledge as potential risk factors, but existing programs do not address these considerations. The proposed research will generate and evaluate the first vaping prevention intervention tailored for teens with CHD. Aim 1: Adapt curriculum from an evidence-based, classroom-delivered vaping prevention intervention to include critical stress management and disease knowledge modules in a mobile Health (mHealth) format for teens (12-18 years) with CHD. Intervention adaptation/refinement will occur via: 1) focus groups with key stakeholders (teens with CHD, parents, cardiologists) to understand intervention preferences, and 2) a single-arm pilot trial of the adapted mHealth intervention (N=16) with follow-up focus groups to gather information for refinement. Aim 2: Evaluate the feasibility/acceptability of the mHealth intervention via a pilot randomized clinical trial (RCT; N=72). Exploratory Aim 3: Explore the preliminary efficacy of the mHealth intervention for indicators of vaping prevention (vaping-related efficacy, perceived vaping outcomes, e-cigarette knowledge). These aims directly align with Dr. Fox’s career goal of becoming an independent clinician-scientist with expertise in behavioral RCTs and scalable disease self-management interventions for transition-aged teens with CHD. To achieve this goal, Dr. Fox requires advanced training in the following areas: 1) behavioral intervention science to design and optimize interventions that can be implemented on a public health level, such as mHealth modalities, 2) RCT methodology and analysis to rigorously evaluate behavioral interventions for efficacy and effectiveness, and 3) CHD disease self- management and healthcare for transition-aged teens to incorporate unique factors of living with CHD into tailored interventions. The career development plan involves didactic and applied training and leverages expert multidisciplinary (public health, clinical/quantitative psychology, cardiology) mentorship from The Abigail Wexner Research Institute at Nationwide Children’s Hospital, one of the worlds’ fastest growing pediatric research centers, its academic affiliate The Ohio State University, and the Fred Hutchinson Cancer Research Center. This K23 proposal will provide high-quality preliminary data and instill expertise needed for an R01 efficacy trial, advancing toward the ultimate goal of enhancing wellbeing across the lifespan of CHD.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY Sleep deficiency remains one of the most prominent and unaddressed public health concerns in pediatric healthcare settings. Pediatric sleep disparities are prominent across minoritized racial subpopulations in the dimensions of sleep duration, timing, alertness, behaviors, and quality/disorders. Despite the evidence of sleep deficiency burdening minoritized youth, these susceptible subpopulations are underrecognized in the clinical workflow leading to sleep medicine specialty services. Ignoring this underlying bias has yielded poorly defined pediatric sleep cohorts in clinical contexts (e.g., historical overrepresentation of White patients). A computable phenotype offers an efficient way to examine a large amount of data from many health systems, specifically electronic health record (EHR) data. Developing a computable phenotype for pediatric sleep deficiency will help us to target sleep screening and care where it is needed the most. However, to do this we will have to ensure the computable phenotype is designed to capture traditionally missed groups and is not biased in a way which harms historically marginalized subpopulations. This K01 will address these equity gaps by identifying potential biases inherent in EHR datasets, understanding their causes, and mitigating them using rigorous methods. The proposed K01 award will allow me to conduct the following aims: 1) the development and validation of a computable phenotype algorithm for classifying pediatric sleep deficiency; and 2) application of postprocessing bias mitigation methods to build and test an equitable computable phenotype model. My primary goal is to become an independent investigator focused on detecting pediatric sleep deficiency and translating that knowledge into effective strategies to improve sleep health in underserved communities. Achieving this goal requires training and research mentorship in specific content areas to (1) learn advanced biomedical informatics approaches for leveraging EHR (e.g., computable phenotyping) and develop an automated screening tool for use by pediatric health systems, (2) develop expertise in population-level sleep disparities research and SDH measurement, and (3) employ responsible conduct of research skills in developing unbiased artificial intelligence (AI) and applying machine learning. My proposed research and training plan will equip me with the skills necessary to become an independent investigator in pediatric sleep research and population health science, prepared to work in interdisciplinary clinical and technical teams. An exceptional interdisciplinary team has been assembled to complete the aims of this K01 research, as well as to mentor me in the training areas critical to my long-term career development. My K01 mentorship team includes both mid-career (Drs. Azizi Seixas, Jennifer Cooper, Christopher Bartlett) and senior mentors/collaborators (Drs. Deena Chisolm, Hongfang Lui, Kelly Kelleher, Lauren Hale), ensuring that I have access to researchers utilizing the latest cutting-edge methods, as well as mentors with large collaborative networks and resources to help launch my career.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT Adolescent cannabis use is a major public health crisis. Marijuana laws (ML) carries significant health impli- cations for American youth. Decriminalization, medicalization, and legalization of cannabis by a majority of US states over the past 25 years has dramatically shifted societal perceptions and adult use patterns. How mariju- ana policy changes have affected population-wide health of US youth and the downstream public health impli- cations of ML remain topics of significant debate. Cannabis is the most commonly used illicit drug by US ado- lescents and the main drug for which US teens obtain substance use (SU) treatment. Adolescent cannabis use is associated with negative long-term consequences on mental health outcomes, risk-taking behaviors (eg, motor vehicle accidents: MVA), and academic/job achievement. Currently, 36 states and the District of Colum- bia (DC) have enacted medical ML (MML); of these, 18 states and DC have enacted recreational ML (RML). While studies have assessed the association between MML and marijuana use in youth, little is known about the impact of ML on mental health outcomes for American youth. No studies to date have evaluated the impact of MML and RML enactment on youth mental health or mortality outcomes. This represents a major knowledge gap in research that, if answered, will provide critical information to guide ML policymaking. The overarching goal of this NIDA RFA-DA-22-037 application is to characterize the effects of state-level MML and RML, including degree of ML restrictiveness and the effect of varying provisions, on changes in can- nabis use disorder (CUD), serious mental illness (SMI), non-cannabis substance use disorder (NCSUD), sui- cide-related outcomes (SROs), all-cause mortality (ACM), and treatment utilization for SU and MH services, including prescription medication, in American youth. Our central hypothesis is that ML will increase CUD, SMI, NCSUD, SROs, and ACM, with less restrictive ML being associated with worse health outcomes. To achieve study objectives, a difference in difference (DID) quasi-experimental design will be implemented. The main in- tervention of interest is ML, specifically, enactment of MML and RML. We will use national Medicaid data from all 50 states and DC for the period 1/1/2008 to 12/31/2020 merged with the National Death Index data. The study population will include a 20% random sample of all US adolescents (12-17 years) and young adults (18- 25 years) (N=55 million). Medicaid-enrolled adolescents and young adults have higher prevalence rates of mental health and substance use disorders and poorer physical and mental health outcomes compared to age matched peers; thus, they are a large and vulnerable population in whom MML and RML effects are unknown. Multilevel models will be fit to obtain estimates of before vs. after changes among adolescents and young adults in states enacting MML and RML compared to changes in other states, controlling for Individual and state-level covariates.

NIH Research Projects · FY 2024 · 2023-07

PROJECT SUMMARY/ABSTRACT PIK3C2B encodes the class II PI3 kinase Phosphatidylinositol-4-phosphate 3-kinase C2 domain-containing beta (PIK3C2B) protein, a regulator of vesicle trafficking through the endosome. Mouse knockout of Pik3c2b has no overtly deleterious phenotype, but instead promotes a beneficial metabolic profile that includes reduced weight gain, improved insulin sensitivity, and, most relevant for this proposal, reduced fat deposition in the liver. In addition, we have previously shown that genetic reduction of Pik3c2b can both prevent and reverse the skeletal muscle phenotype, as well as extend survival, of the mouse model of X-linked myotubular myopathy (XLMTM). XLMTM is rare neurogenetic condition with onset in infancy that is associated with profound muscle related disabilities and early death. In addition to affecting the skeletal muscle, an emerging XLMTM phenotype is hepatobiliary disease, which progresses to liver failure in a subset of patients. There are no treatments for XLMTM. Importantly, liver disease has not previously been identified in pre-clinical models of XLMTM, a significant barrier for disease understanding and treatment. To overcome this barrier, in new data, we have developed the first mouse model of XLMTM liver disease. Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent condition that affects nearly 25% of American adults. It can lead to non-alcoholic hepatosteatosis (NASH), which in turn can progress to cirrhosis, liver failure, liver cancer, and death. There are no adequate therapies for these common and severe conditions. The scientific premise of this proposal is that reduction of PIK3C2B in mice promotes reduction of diet induced fatty liver and amelioration of disease relevant XLMTM phenotypes. The overall goal of the project is to develop therapies for XLMTM and NAFLD/NASH using a RNA knockdown based gene therapy. We hypothesize the following: (1) PIK3C2B knockdown in muscle and liver can be specifically, lastingly, and safely achieved with a synthetic miRNA delivered in vivo with AAV8 (AAV-miR-PIK3C2B); (2) AAV-miR- PIK3C2B can ameliorate both muscle and liver and phenotypes of the XLMTM mouse model; and (3) AAV-miR- PIK3C2B can prevent the development and progression of diet induce mouse NAFLD/NASH. These hypotheses will be rigorously evaluated in 3 aims. Aim 1 will develop AAV-miR-PIK3C2B, Aim 2 will test it in XLMTM mice, and Aim 3 will study it in a diet induced model of NASH. If successful, this proposal will be of high significance and impact because it will identify a single therapy suitable for clinical translation for both a rare, fatal paediatric condition and for a prevalent disease affecting millions of adults.

NIH Research Projects · FY 2024 · 2023-07

Project Summary Hypersomnolence (HYP), or excessive daytime sleepiness, is the most common symptom encountered in sleep medicine, and can present as linked to other medical disorders or independently. Discriminating among the multiple possible causes of HYP is a complex process, and the underlying cause is often unknown. Furthermore, there are currently no reliable electrophysiological parameters or biomarkers for HYP, which is a severe limitation to the diagnostic and therapeutic process. Understanding the biophysical presentation of HYP in sleep brain dynamics is essential to both the identification of reliable electrophysiological biomarkers and to building a mechanistic understanding of the physiological manifestations of HYP. Most studies of sleep EEG dynamics focus on rhythms uniformly grouped by their dominant frequency, sometimes addressing their spatial presentation, but overall ignoring the articulation of sleep rhythms in space-time organized events. In recent work on typical adult populations the PI has introduced data- driven techniques that reveal the space-time patterns of slow oscillations (SOs) and spindles, both sleep rhythms cardinal to sleep homeostasis, with SOs explicitly tied to the restorative-ness of a night of sleep. This research line has also shown that differentiation of sleep rhythms in space-time patterns is a powerful approach to revealing biophysical differentiation among events classified as the same “rhythm” suggesting their potential differential contribution to sleep functions. Here, we propose to apply these data-driven approaches to describe in detail the space-time presentation of HYP in sleep brain dynamics, in order to determine HYP biomarkers and to advance our understanding of the manifestations of HYP in brain activity important to health and cognition. This study will re-analyze a well-characterized dataset including the sleep studies of persons with HYP and controls, with both groups also articulated based on presence/absence of major depressive disorder. Specifically, we will describe the space-time patterns of SOs and spindles on the scalp and their main biophysical properties, comparing them among the HYP and control group (aim 1). We will then use machine learning classification to determine for each individual the estimated cortical-subcortical currents that most differentiate SOs space-time types, compare the results in HYP and controls (aim 2). Finally, we will statistically evaluate the link between these biophysical quantifiers of space-time sleep patterns and clinical/behavioral assessments of HYP symptoms, depression, and anxiety. This research will lead to new insights into potential brain mechanisms that underlie HYP, as well as refined diagnostic and future therapies for the multitude suffering with HYP disorder.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Low-grade gliomas (LGGs), a heterogeneous group of primary central nervous system tumors, are one of the most common solid tumors in the adolescent and young adult (AYA) population. LGGs can progress to high- grade gliomas (HGGs) via a process known as malignant transformation (MT), resulting in dismal prognoses. The mechanisms driving MT of LGG remain poorly understood. In contrast to LGGs, we and others have shown that HGGs have increased levels of bone marrow-derived myeloid cells (BMDMs) and myeloid-derived suppressor cells (MDSCs) in peripheral circulation, with an intra-tumoral enrichment of tumor-associated macrophages (TAMs) and a paucity of CD8+T and natural killer (NK) cells. Although myeloid cells are known to accumulate during glioma progression, it remains unclear if these cells have a causal role in driving MT. Central hypothesis: Reductions in anti-tumor reactive CD8+T and NK cell-dependent immune surveillance are responsible for LGG to HGG malignant transformation in AYA patients and these reductions are dictated by increased infiltration and immune suppressive activity of tumor infiltrating BMDMs. We have identified two independent myeloid associated pathways that are responsible for these BMDM phenotypes and we propose to test the central hypothesis through the following two aims: Aim 1. Delineate the dependency of BMDM cell differentiation on inhibitor of DNA binding protein 2 (ID2). Using AYA RCAS/tv-a glioma mouse models, we will knockdown ID2 in BMDMs before and during MT. We will examine effects on pro-tumoral myeloid cells and evaluate intra-tumoral T and NK cells and mobilization in blood, spleen, and bone marrow at various times with scRNAseq, mass cytometry, and fluorescent-activated cell sorting. We will also leverage digital spatial transcriptomics (DSP) in ID2 modulated tumors and validate ID2 signaling in human paired LGG /HGG samples to dissect the expression of myeloid, NK, T cell markers, and checkpoint molecules to illuminate ROIs critical to MT. Results will illuminate ID2 mechanisms of BMDM cell differentiation to pro-tumoral myeloid cells. Aim 2. Determine if the CD74/macrophage migration inhibitory factor (MIF) axis regulates BMDM cell differentiation during LGG malignant transformation. We will investigate CD74’s role in MT using pharmacological treatments or transplant wild-type or CD74 KD/KO bone marrow cells in a murine RCAS/tv-a glioma model. We will analyze peripheral blood and the glioma TME using scRNAseq, mass cytometry, and FACS. We will validate findings with bulk RNA sequencing data on paired LGG/HGGs from AYA patients and investigate how CD74/MIF signaling drives reduced T and NK cells which supports LGG MT. We will also leverage DSP in CD74/MIF modulated tumors and validate CD74 signaling in human paired LGG/HGG samples to dissect the expression of myeloid, NK, T cell markers, and checkpoint molecules to illuminate ROIs critical to MT that will indicate potentially targetable vulnerabilities. Overall Impact: Our studies will illuminate novel insights that may enable us to translate into enhanced immune surveillance approaches to delay and/or prevent MT for AYA LGG patients.

NIH Research Projects · FY 2024 · 2023-07

Abstract Disease of the heart, lungs and blood are associated with substantial morbidity and mortality in pediatric populations. Children make up 22% of the U.S. population, yet the NIH’s $4 billion pediatric research portfolio makes up only 10% of the annual NIH research budget. The Nationwide Children’s Hospital Research Experiences in Childhood Heart, Lung and Blood Sciences (REaCH-LABS) is designed to provide meaningful summer opportunities for undergraduate students from backgrounds underrepresented in medicine (URiM) in scientific areas of interest to NHLBI. Each annual cohort will include 5 new participants who are eligible for up to two years in the program (yielding a total of 10 participating students each year). Our exceptional research mentorship faculty, representing diversity in both science and demographics, is comprised of over 20 PIs with current or past NHLBI funding or other relevant federal funding. Participants will be situated within a dynamic ecosystem of pipeline programs at Nationwide Children’s Hospital spanning high school research internships thoughT32-funded post-doctoral programs. The curriculum will include 1) an immersive summer research experience in a research team conducting NHLBI priority research; 2) assignment of a near-peer (medical or graduate student) mentor to support readiness for transition to graduate or medical school; 3) weekly Heart of the Matter research seminars providing exposure to active research projects and clinical careers; and 4) weekly professional development workshops focused on medical school applications, MCATs, interviewing and other relevant topics. Research team experiences will include meaningful mentored projects that build skills and produce a product suitable for presentation at the close of the internship. Highly qualified internal and external review boards will provide guidance on implementation and outcomes and a structured approach to follow-up will tracking of long-term program outcomes. To maximize our pool of well-qualified URiM candidates, this project includes a novel partnership with the Student National Medical Association (SNMA), the nation’s largest organization supporting African American medical students, and its undergraduate affiliate organization, the Minority Association of Pre-Health Students (MAPS). SNMA leadership and members will engage in project oversight, participant recruitment, near-peer mentoring, and panel presentations, serving as both contributors and role models. REaCH-LABS is committed to recruiting, engaging, and supporting URiM participants in a structure that supports their oft-times unique needs in a way that smooths their path to a biomedical research career. Our program design will generate well-qualified graduates who add to the diversity of researchers and research questions, promoting health equity for children and the adults that they will become.

NIH Research Projects · FY 2025 · 2023-07

The objective of this Nationwide Children’s Hospital (NCH)/Ohio State University (OSU) institutional T32 postdoctoral program is to provide interdisciplinary, state-of-the-art research training in pediatric heart and lung diseases. Cardiopulmonary diseases in infants and children are among the most severe type of pediatric illnesses ranging from congenital malformations and monogenic conditions to acquired infectious/inflammatory diseases. Our program provides a unique integrated approach to prepare trainees to become scientists and physician-scientists who will make basic, translational, and clinical research discoveries through cooperative, collaborative team science. The rationale for this holistic approach, which spans from molecular discovery and translational approaches using cell and animal models to human clinical trials, is that these synergistic methods will more rapidly impact patient care and therapies for children with cardiopulmonary diseases. We will provide an academic environment that cultivates research training at a leading pediatric research institute affiliated with an internationally recognized children’s hospital and university. Dynamic ‘bidirectional’ research tracks will allow for clinical problems to be mechanistically studied by physician-scientists and new, discovery-based findings of fundamental disease mechanisms in basic research labs to be effectively translated to the clinic. This application seeks to support 4 trainees each year over the funding period. Fellows will be selected from MD, DO, MD/PhD, or PhD trainees who hold strong interests in pediatric cardiopulmonary research and are committed to careers in disease-oriented investigation. The Program Directors have strong records in basic, translational, and clinical cardiopulmonary research and mentoring, and will serve as the training program leaders. The training plan consists of laboratory and clinical research training supplemented by courses, seminars, conferences, and exposure to industry relationships and non-traditional career pathways. Each trainee will perform independent research under the direction of a mentor according to an individualized mentoring program. The research areas encompassed by the 27 mentors, including 6 Mentors-in-Training, are represented by their affiliations at the Abigail Wexner Research Institute at NCH and The Ohio State University. Overall metrics to determine success include programmatic review by internal and external advisory committees and the successful execution of each trainee’s individual mentoring program. Trainee-specific metrics include required coursework, mentoring team evaluations, and research productivity as measured by publications, presentations, and grants. A comprehensive trainee and mentor evaluation system, an integral component of the training program, is also included.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY/ABSTRACT Osteosarcoma and Ewing sarcoma are the most common pediatric bone malignancies and efforts to treat patients with advanced disease remain dismal, despite decades of research. To address this inadequacy, we seek to leverage oncolytic virotherapy as a twofold attack against tumors: direct tumor cell lysis and antitumor immune stimulation. However, previous trials of oncolytic herpes simplex virus (oHSV) in pediatric sarcomas displayed limited responses, likely due to immunosuppressive monocytes and macrophages. To improve efficacy, we combined oHSV with trabectedin, an FDA-approved DNA-binding agent known to mitigate monocytes and macrophages. Strikingly, we found that the efficacy of this combination far surpassed our expectations, inducing complete regressions and increased survival in multiple models of Ewing sarcoma and osteosarcoma. The mechanism for this synergy remains unknown and understanding it is critical to optimize approaches to therapy in human trials. In our preliminary single-cell RNA sequencing data investigating this synergy, we observed far more viral transcripts in combination-treated tumors than in oHSV-treated tumors and a decrease in the expression of genes associated with the intrinsic antiviral response, suggesting that trabectedin selectively sensitizes tumor cells to infection. We also found that oHSV induced increased TRAILR2 death receptor expression in tumor cells and increased TRAIL expression in NK cells, pointing to a second potential mechanism for synergy through cytotoxic signaling. Based on these data, we hypothesize that trabectedin augments oHSV efficacy through mechanisms that include transcriptional inhibition of tumor-intrinsic antiviral responses and enhanced NK-mediated cytotoxicity via TRAIL-TRAILR2 signaling. Firstly, we will investigate the decreased intrinsic antiviral response and the oHSV-induced tumor cell death pathways under combination therapy via proteomic methods, including mass cytometry (CyTOF), western blot, and multi-color immunofluorescence. To observe the related tumor cell sensitivity to oHSV, we will construct a luminescent oHSV and monitor its spread in real-time. Secondly, we will analyze TRAIL, TRAILR2, TRAILR2 inhibitors, and caspase-8 pathway expressions using CyTOF, western blot, ELISA, multi-color immunofluorescence, and qPCR, in combination-treated tumors compared to oHSV-treated tumors. To determine whether TRAILR2 signaling is necessary for combination efficacy, we will use CRISPR-Cas9 to knock out tumor cell TRAILR2 and compare single and combination treatment efficacies in wild-type and knock-out models. Ultimately, we will illuminate mechanisms of synergy through a two-pronged analysis, with the potential to reveal numerous generalizable vulnerabilities in osteosarcoma and Ewing sarcoma for clinical application and the development of future synergistic treatments against pediatric sarcomas.

NIH Research Projects · FY 2026 · 2023-04

(<30 lines) Youth-onset type 2 diabetes (YO-T2D) is increasingly prevalent in parallel with the obesity epidemic, yet effective treatment and prevention strategies are limited. The physiologic increase in insulin resistance occurring during puberty, in combination with obesity-related insulin resistance, enhances the risk of T2D. Yet, it remains unclear why some youth progress through puberty with intact β-cell function, while others do not, despite similar phenotypic and metabolic characteristics. More information is needed regarding the unique events during puberty to better understand 1) the basic pathophysiology of glucose control, insulin sensitivity, β-cell function, and T2D risk in youth, 2) differences among girls and boys, populations at highest risk, and urban and rural geographies, and 3) the potential contribution of other risk factors including psychological, behavioral, and social and external contexts. Importantly, this research needs to address the timeline of pathophysiology and progression from normoglycemia or prediabetes to YO-T2D. The DISCOVERY of Risk Factors for Type 2 Diabetes in Youth (DISCOVERY) study provides a unique opportunity to characterize the risk progression profile and mechanisms underlying the development of YO-T2D, and evaluate the effects of modifiable and non-modifiable risk factors. Ultimately, the results of this study will establish a basic pathophysiology to inform future studies aimed at achieving target glycemia, improving insulin sensitivity, preserving β-cell function, and/or preventing YO-T2D. To address this goal, DISCOVERY will recruit, enroll, and follow a nationally-representative cohort of 3,600 at-risk obese youth in early puberty; extensively phenotype them as they transition through puberty; and characterize the course of decline and dysfunction in pathophysiological indicators that lead to YO-T2D. The expected duration of the DISCOVERY is 5 years, including planning, recruitment, follow-up, analysis, and reporting. In addition, DISCOVERY will store longitudinal biospecimens and genetic material with the intention of acquiring additional ancillary funding to pursue analysis of emerging indicators. Nationwide Children's Hospital has experience in multicenter and diabetes-related investigations and will contribute to DISCOVERY through the recruitment of approximately 240 at-risk youth, implementation of the IRB-approved consensus protocol, participation on DISCOVERY committees, and collaboration on the analyses and dissemination of the findings from DISCOVERY.

NIH Research Projects · FY 2026 · 2023-04

Summary The goal of the proposed study is to better understand the role of antibodies in HEV infection and determine if antibodies can prevent or cure chronic hepatitis E virus (HEV) infection. HEV infections are usually self-limited, but the infections frequently persist when the immune system is compromised and if left untreated, can lead to serious liver disease. HEV exists in two distinct virion forms: naked virions (nHEV) that are shed into feces and mediate virus transmission between hosts, and quasi-enveloped HEV (eHEV) virions that circulate in the bloodstream and mediate virus spread between cells. The eHEV particles lack viral antigens on their surface, thus they are resistant to circulating HEV-specific antibodies. We previously show that eHEV particles enter cells via a novel entry mechanism that involves lysosomal degradation of the viral envelope. Our recent data show that HEV-specific IgG, but not IgM, effectively block eHEV-mediated spread in cell culture. Our central hypothesis is that antibodies neutralize eHEV intracellularly by preventing virus uncoating in the endosome/lysosome where the viral membrane degrades. Antibodies generated by natural HEV infection and vaccination with truncated HEV capsid proteins (CP) are highly protective against HEV infection, while anti-HEV antibody titers are usually low in patients with chronic HEV infection. Thus, antibodies may have the potential to prevent or treat chronic HEV infection. Despite these encouragements, there are several significant roadblocks. First, the C terminus of the HEV CP, which is not present in the current vaccine and the fecal virus, is intact in the eHEV particles. This is important since structural modeling suggests that the presence of the C terminus of CP significantly alters the surface structure of the virion which likely makes vaccine-induced antibodies less effective against eHEV. Second, our recent work indicates that HEV produces a capsid decoy that is secreted from infected cells in a large quantity and interferes with antibody-mediated neutralization. Third, antibody uptake by hepatocytes is an inefficient process. Here we propose to overcome these obstacles. Aim 1 will test the hypothesis that antibodies targeting virions with intact CP will block eHEV-mediated spread more efficiently. We will determine the structure of authentic HEV virions with intact or cleaved CP and assess if antibodies targeting virions with intact CP neutralize eHEV more efficiently. We will also determine if glycoengineered antibodies with enhanced lysosomal targeting neutralize eHEV more efficiently. Aim 2 will test the hypothesis that neutralizing antibodies that do not bind or bind poorly to the decoy will block HEV spread more efficiently. We will also determine the structure of the CP decoy in complex with antibodies by cryo-EM to gain a better understanding of the evasion mechanism by the decoy. The completion of the proposed work will provide novel insights into the role of antibodies in HEV infection and inform strategies to prevent or cure chronic HEV infection.