Children'S Hosp Of Philadelphia

NIH Research Projects · FY 2025 · 2023-04

Rett syndrome (RTT), is a severe neurodevelopmental disorder caused by loss-of function mutations in the X- linked gene Methyl-CpG-binding Protein 2 (MECP2) and characterized by loss of speech and hand skills, problems walking, and repetitive hand movements. Genetic restoration of MECP2 in symptomatic mice can reverse symptoms providing hope that disease-modifying therapies can be created. Impeding the development of transformative therapies are a lack of biomarkers of treatment-response. Ideally, a biomarker can be applied in mice and humans to enhance effective translation of preclinical treatment studies and improve human trial design and execution. Neurophysiological assessments have potential as biomarkers as they are non-invasive, measure neurological changes, and are translatable between humans and animal models. Recent work in RTT, from our group, has found differences in neurophysiological measures in both affected humans and mouse models that correlate with disease severity, but an urgent need exists to identify well-validated and translatable treatment-response biomarkers in RTT. To address this need, we propose here to develop neurophysiological biomarkers that can fulfil a specific primary Context of Use (COU), an early treatment response biomarker, to facilitate and speed both preclinical and clinical trials of novel therapies in RTT. The primary goal of the R61 phase of the proposal is to identify candidate neurophysiological biomarkers of disease improvement in a mouse model of RTT and establish human multi-site standard operating procedures and normative data. These parallel projects will be foundational to identify a true treatment responsive biomarker in RTT. To do this we will first determine if potential biomarkers, quantitative EEG and evoked potentials will change predictively in a mouse model of RTT that allows for genetic rescue of the RTT phenotype. Simultaneously, we will develop and optimize standard operating procedures to enable multi-site evaluation of candidate human neurophysiological biomarkers. Additionally, we will evaluate test-retest reliability of the biomarkers we are developing. Finally, we will determine if the putative neurophysiological biomarkers change during active clinical change in RTT. For the R33 phase, we will demonstrate that our human proof-of-concept of candidate neurophysiological biomarkers are stable over the time frame relevant to clinical trials in RTT and that these biomarkers correlate with RTT clinical severity. Overall, this proposal takes advantage of the ability to use mouse models to identify and validate robust human neurophysiological features as putative biomarkers. These neurophysiological measures will allow for accelerated therapy development via the replacement of subjective clinical findings with quantitative measures of early treatment-response. Together, this work will facilitate biomarker development to be employed in interventional therapy development.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Type 1 diabetes (T1D) is an autoimmune disease that affects millions of people worldwide. The incidence of T1D is rising, especially in young children. Although significant progress has been made to predict who is at risk for developing T1D, there are no effective therapies to prevent this disease. Both genetic and environmental factors contribute to the risk of developing T1D. Certain major histocompatibility complex/human leukocyte antigen (MHC/HLA) class II haplotypes dominantly protect against the development of T1D, and we recently discovered that protective MHCII molecules shape early-life microbial communities which in turn impact immune system development to prevent T1D. Modeling microbial protection from T1D in NOD mice may provide critical insights to support our long-term goal of developing microbiota-based therapies to prevent T1D in humans. Due to the complexity and high levels of variability of the intestinal microbiome, determining the specific microbial strains that drive immune system development and function is problematic. The development of gnotobiotic mice with defined adult microbial communities has been an important advance in the field because they simplify the complexity and variability of the system and allow for well-controlled, mechanistic studies. However, a gnotobiotic mouse model to study pediatric disease is lacking. We developed a new gnotobiotic mouse model of the early- life microbiome which we call Pediatric Community or “PedsCom”. PedsCom is a consortium of 9 bacterial strains isolated from the intestine of pre-weaning diabetes-protected Eα16/NOD mice. Remarkably, this 9- microbe community robustly induces regulatory T cells (Tregs) and confers protection from T1D to diabetes- susceptible NOD mice. We hypothesize that specific PedsCom microbes work in concert to prevent T1D by providing microbial antigens and metabolites that induce peripheral regulatory T cells (pTregs) during a critical early life window of immune system development. Aim 1 examines the timing, localization, and metabolites produced by specific PedsCom members which drive pTreg cell development and prevent autoimmunity. Aim 2 examines the mechanisms by which pTregs are induced by PedsCom microbes and their protein antigens and whether pTregs whose TCRs recognize specific microbial antigens mediate protection from T1D. Successful completion of these aims will provide critical information on which early-life microbes induce pTregs, and the degree to which microbial antigens and metabolites work together to generate a diabetes- protective immune system. In addition, PedsCom mice are an innovative tool for investigating early-life host- microbiota interactions.

NIH Research Projects · FY 2026 · 2023-04

Rett syndrome (RTT), is a severe neurodevelopmental disorder caused by loss-of function mutations in the X- linked gene Methyl-CpG-binding Protein 2 (MECP2) and characterized by loss of speech and hand skills, problems walking, and repetitive hand movements. Genetic restoration of MECP2 in symptomatic mice can reverse symptoms providing hope that disease-modifying therapies can be created. Impeding the development of transformative therapies are a lack of biomarkers of treatment-response. Ideally, a biomarker can be applied in mice and humans to enhance effective translation of preclinical treatment studies and improve human trial design and execution. Neurophysiological assessments have potential as biomarkers as they are non-invasive, measure neurological changes, and are translatable between humans and animal models. Recent work in RTT, from our group, has found differences in neurophysiological measures in both affected humans and mouse models that correlate with disease severity, but an urgent need exists to identify well-validated and translatable treatment-response biomarkers in RTT. To address this need, we propose here to develop neurophysiological biomarkers that can fulfil a specific primary Context of Use (COU), an early treatment response biomarker, to facilitate and speed both preclinical and clinical trials of novel therapies in RTT. The primary goal of the R61 phase of the proposal is to identify candidate neurophysiological biomarkers of disease improvement in a mouse model of RTT and establish human multi-site standard operating procedures and normative data. These parallel projects will be foundational to identify a true treatment responsive biomarker in RTT. To do this we will first determine if potential biomarkers, quantitative EEG and evoked potentials will change predictively in a mouse model of RTT that allows for genetic rescue of the RTT phenotype. Simultaneously, we will develop and optimize standard operating procedures to enable multi-site evaluation of candidate human neurophysiological biomarkers. Additionally, we will evaluate test-retest reliability of the biomarkers we are developing. Finally, we will determine if the putative neurophysiological biomarkers change during active clinical change in RTT. For the R33 phase, we will demonstrate that our human proof-of-concept of candidate neurophysiological biomarkers are stable over the time frame relevant to clinical trials in RTT and that these biomarkers correlate with RTT clinical severity. Overall, this proposal takes advantage of the ability to use mouse models to identify and validate robust human neurophysiological features as putative biomarkers. These neurophysiological measures will allow for accelerated therapy development via the replacement of subjective clinical findings with quantitative measures of early treatment-response. Together, this work will facilitate biomarker development to be employed in interventional therapy development.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY/ABSTRACT This K08 Award encompasses a research and training plan to facilitate Dr. Regina Myers' transition to an independent clinical investigator. Dr. Myers is currently an Instructor of Pediatrics and a pediatric oncology and cell therapy physician at Children's Hospital of Philadelphia and the University of Pennsylvania. Her long-term goal is to develop an independent research program that will integrate early phase clinical trials with advanced epidemiology methodologies in order to improve outcomes for children and young adults with high-risk hematologic malignancies. The training objectives for this award will bridge her prior experience in immunotherapy and outcomes research to her long-term goals, and include: acquiring independence in the design and implementation of early phase immunotherapy clinical trials, expanding her expertise in clinical epidemiology to include advanced methods for causal inference, establishing proficiency in the assessment of clinical trial correlative endpoints, and gaining experience with the application of synthetic control arms in pediatric cancer clinical trials. The proposed activities will be conducted in the resource-rich environment at CHOP/Penn and under the mentorship of an expert, multidisciplinary team led by Dr. Stephan Grupp, an international leader in cancer immunotherapy. CD19-specific chimeric antigen receptor-modified T cells (CAR19) have demonstrated unprecedented responses in relapsed or refractory B-cell acute lymphoblastic leukemia. Unfortunately, however, 50% of children and young adults relapse suffer a subsequent relapse and their prognosis after post-CAR19 relapse is dismal. As the use of CAR19 broadens, there is a corresponding increase in the number of patients with post-CAR19 relapse, creating a critical need to identify optimal salvage treatment approaches. The proposed research aims to improve outcomes after post-CAR19 relapse using novel and complementary clinical research approaches. In Aim 1, Dr. Myers will perform a phase 1/2 clinical trial to test a dual-antigen targeted CAR designed to overcome the primary mechanisms of CAR failure in children with post-CAR19 relapse. Secondary analyses will evaluate the predictive value of specific biomarkers for CAR failure and will compare the dual-targeted CAR against synthetic external control data. In Aim 2, Dr. Myers will assess the comparative effectiveness of existing treatment approaches for post-CAR19 relapse using randomized clinical trial emulation methods, leveraging clinical trial and real-world data from the largest, single-center pediatric CAR cohort. Successful completion of this career development award will advance the field through the introduction of a novel, dual-antigen targeted CAR and the establishment of a robust clinical research infrastructure capable of integrating real-world data with clinical trial data to determine optimal existing treatment strategies and efficiently assess new therapies as they become available. The proposed studies and training plan will provide an outstanding foundation for Dr. Myers' career as a physician-scientist.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Neuronal or axonal damage in the central nervous system (CNS), caused by injury or diseases, is irreversible and may lead to persistent neurological deficits. Spinal cord injury (SCI) often causes severe sensory and motor dysfunction and paralysis. Of approximately 1.9% of the U.S. population living with paralysis, over 1,275,000 are paralyzed as the result of SCI. Currently, there is still no cure for the injured spinal cord itself, emphasizing the desperate need to identify novel pathways for targeted therapy. Regarded as the holy grail in regenerative medicine, achieving axon regeneration and functional recovery after CNS injury or in neurodegenerative diseases remains a daunting task. The inability of CNS axons to regenerate after injury is attributed to the reduced intrinsic growth capacity of neurons and the inhibitory milieu largely constituted by the reactive glial cells. It is conventionally thought that the structural formation of glial scar and its upregulation of the repulsive CSPGs are the main culprit leading to stalled regrowth. However, accumulating evidence in the past decade has demonstrated that preventing astroglial scar formation following CNS injury does not result in increased regrowth. It is proposed that glial scar is important in preserving tissue integrity and mitigating further inflammatory damage. Glial scar may have beneficial effects during the acute phase of injury, but prevents axon regrowth in the chronic or later stages. In our latest work, via glia-specific metabolic reprogramming, we succeeded in mitigating their adverse effects while enriching their promotive functions. We demonstrated that glial reprogramming enhances glial glycolysis, and the production and release of metabolites – lactate and L-2HG, which act through neuronal GABABRs to boost axon regeneration. However, major gaps remain: are lactate and L-2HG the only pro-regeneration metabolites; do anti-regeneration metabolites also exist; do glia subtypes behave similarly after metabolic reprogramming. Our published work allows us to ask the essential question: does metabolic status dictate glia’s ability to promote or inhibit CNS axon regeneration? This would have a fundamental impact on our understanding of axon regeneration, as it applies to all species across the evolution spectrum. An equally intriguing question is: does the metabolic status differ between regeneration competent and incompetent CNS neurons? Our proposal aims to answer these questions, and test our hypothesis that glial and neuronal metabolic status governs the regeneration capacity of CNS neurons. Although various strategies to boost the neuronal intrinsic regenerative ability, to remove the extrinsic inhibitory factors such as CSPGs, to transdifferentiate glia into neurons, or to transplant stem cells into CNS have been reported, none of them have translated into clinical use. There is still a pressing need for new concepts to promote CNS axon regeneration. Our pilot results demonstrate that the state of glial cells that promotes axon regeneration can be achieved by reprogramming. This project aims to uncover metabolic enzymes as therapeutic targets, and metabolites or their derivatives as potential pharmacological agents for treating CNS injury.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Over the last decade, there has been an exponential increase in identified genetic causes of neurodevelopmental disorders and epilepsy. With more than 100 genes identified, understanding how phenotypes relate to specific genetic variants is critical, given the clinical complexity of developmental brain disorders. Given that treatment and prognosis is dependent on understanding genotype-phenotype correlations, there is a critical need to better assess clinical features in genetic epilepsies. However, phenotyping is a time-consuming, manual task with limited throughput. To overcome this bottleneck, we have developed a novel approach, based on the Human Phenotype Ontology (HPO), which we have previously applied to SCN2A-related disorders and to STXBP1- related disorders, resulting in knowledge that is already applied clinically. Our long-term goal is to decipher the phenotypic landscape of genetic epilepsies to improve clinical care.Therefore, our objectives are to determine the relationship between genomic variation and epilepsy-related clinical features in a large patient cohort and to identify subgroups within the 20 most common genetic epilepsies that may provide insight into outcomes and treatment responses. We plan to pursue these objectives through two aims. First, we aim to determine the impact of genomic features on epilepsy phenotypes in >9,000 individuals through an HPO-based approach (Aim #1). We will analyze exome data in >13K individuals with trio exome data and >600K HPO terms to assess the relationship between distinct monogenic etiologies and rare variants with clinical epilepsy features, using computational phenotyping tools developed by our team. This will allow for insight into the relationship between genetic etiologies and phenotypic features at a granular scale. Secondly, we aim to define relevant subgroups in genetic epilepsies through phenotype harmonization (Aim #2). We will translate clinical features for the 20 most common genetic epilepsies to HPO terms and perform a semantic similarity analysis to determine whether specific variants have significantly similar clinical features, followed by in-depth chart review. This knowledge will inform the prioritization of variants for functional studies and clinical care. In summary, HPO-based delineation of genetic epilepsies is expected to significantly improve knowledge of genotype-phenotype correlations by adding unmatched detail and power. Our team has previously pioneered computational phenotype analysis in the epilepsies and neurodevelopmental disorders, positioning us uniquely to address these questions. In addition to facilitating research of disease mechanisms by prioritizing variants for work with stem cells or mouse models, for example, our findings will also apply to clinical care by providing an unprecedented level of precision in prognosis and treatment information.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Over the last decade, there has been an exponential increase in identified genetic causes of neurodevelopmental disorders and epilepsy. With more than 100 genes identified, understanding how phenotypes relate to specific genetic variants is critical given the clinical complexity of developmental brain disorders. Given that treatment and prognosis is dependent on understanding genotype-phenotype correlations, there is a critical need to better assess clinical features in genetic epilepsies. However, phenotyping is a time-consuming, manual task with limited throughput. To overcome this bottleneck, we have developed a novel approach, based on the Human Phenotype Ontology (HPO) to capture and analyze longitudinal phenotypic data. In our preliminary data for STXBP1- and SCN8A-related disorders, which we have reconstructed for >550 patient months, we identified unique natural histories, outcomes, and distinct response patterns to specific treatment strategies. Natural history and treatment response in pediatric epilepsies are deeply intertwined and often difficult to disentangle. Accordingly, a comprehensive assessment of genetic epilepsies needs to account for two factors, subgroups with common clinical trajectories as well as gene-specific treatment responses. Our suggested project therefore has two aims. First, we plan to detect relevant subgroups in genetic epilepsies based on longitudinal clinical data (Aim #1). We will reconstruct longitudinal trajectories and outcomes in the 15 most common genetic epilepsies with 50-75 individuals per gene to delineate longitudinal seizure burden, seizures types, and developmental milestones. Based on this, we will then identify subgroups defined by clinical features and global clinical resemblance, as well as variant and gene groups. In addition (Aim #2), we will identify specific treatment responses in genetic epilepsies using standardized phenotypes. We will combine reconstructed natural history with treatment data to compare reduction in seizure frequencies and effect on maintaining seizure freedom across >20 treatment strategies with the goal to identify the most effective treatment strategy when adjusting for age and seizure type. Finally, we will also compare medication response across major variant classes and across all genetic etiologies combined. Our team has previously pioneered computational phenotype analysis in the epilepsies, positioning us uniquely to address these questions. Our analysis, mapping longitudinal clinical data to a harmonized format will provide unprecedented granularity in deciphering the trajectory of genetic epilepsies, informing clinical practice in these conditions. We hope that these results will provide a template for the analysis of the limited clinical data in rare diseases in order to maximize treatment-relevant information, especially in conditions with complex, longitudinal disease histories.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY/ABSTRACT Obesity causes the greatest proportional risk for diabetes, heart disease, and cancer and is resistant to current treatments despite being a focus of intense research. Obesity occurs because the body stores surplus calories as fat, which in turn drives the health risks associated with obesity6-10. The current therapeutic approaches to obesity focus on weight loss, via caloric restriction and/or exercise, which are not effective. An alternate strategy would be to redirect surplus calories to build muscle instead of storage as fat. This approach would mitigate the health risks of obesity and also improve daily functioning, quality of life, and longevity. Our intriguing preliminary data reveal that high-dose dietary vitamin D decreases the proportion of excess calories stored as fat, instead allocating these calories to muscle. Understanding the mechanisms underlying this finding will drive the development of novel therapeutic approaches desperately needed to prevent and treat obesity. Further, our preliminary results suggest that this calorie allocation to muscle occurs via 25D mediated vitamin D receptor (VDR) transcriptional regulation at non-canonical VDR binding sites. Our long term goal is to define the roles of vitamin D in calorie allocation in order to identify novel therapeutic targets in obesity. The specific objectives of this project are 1) to determine which dietary vitamin D metabolite, 25D or 1,25D, signals to allocate calories to muscle, and 2) to identify the mechanisms in muscle whereby vitamin D signaling leads to changes in gene expression underlying calorie allocation. Using mouse models, we have demonstrated that high-dose dietary vitamin D increases muscle mass, cross sectional area, strength/area, and muscle mitochondrial capacity in both lean and obese mice. Our central hypothesis is that high-dose vitamin D calorie allocation is mediated by 25D acting via the VDR to alter transcription through non-canonical binding sites. Our approach uses validated genetically engineered mouse models of vitamin D imbalance, and connects signaling to transcriptional changes by genome-wide analysis of VDR binding. In sum, this proposal describes a five-year research plan to understand the mechanisms underlying non- calciometabolic actions of vitamin D to preferentially allocate calories to muscle instead of fat with the long-term goal of developing novel rational therapies for obesity. The primary investigator is an Assistant Professor on the tenure track at the University of Pennsylvania. He is an early career researcher dedicated to asking translational questions to better understand non-calciometabolic actions of vitamin D. He has assembled a uniquely qualified and complementary collaborative team to tackle the objectives of this application. Successful completion of this work will define the role of 25D in mediating vitamin D muscle calorie allocation and will identify relevant signaling pathways that could be targeted therapeutically to preferentially allocate surplus calories to muscle instead of fat, thereby decreasing both unwanted effects and incidence of obesity.

NIH Research Projects · FY 2026 · 2023-02

Technical abstract Genetic study of orofacial clefts (OFC) is foundational to genetics of congenital structural birth defects. Most OFC cases are non-syndromic and involve complex genetic mechanisms that are yet to be fully elucidated. Currently, genetic diagnosis for cleft anomalies is hampered by two critical knowledge gaps. First, genes essential for palate formation are incompletely identified. Second, even when the cleft risk genes are associated, algorithms used to impute deleterious from benign gene variants via computational and statistical methods remain unreliable. There is a critical need to translate genome sequencing to clinically actionable data, where functional studies provide the highest-level evidence to impute pathogenicity. We showed that Esrp1 and its paralog Esrp2 (hereafter Esrp1/2) operate in the periderm of mouse and zebrafish to regulate craniofacial development. Esrp1/2 mediates alternative splicing of RNA transcripts, creating epithelial isoforms that function in oral epithelium and periderm. This proposal tests the central hypothesis that Esrp1/2 is required to generate epithelial isoform of Ctnnd1, which maintains periderm integrity necessary for craniofacial morphogenesis. We will impute pathogenicity of ESRP1/2 and CTNND1 human gene variants associated with OFC. Using completed genome sequencing projects and projects in progress, we curate large numbers of ESRP1, ESRP2 and CTNND1 gene variants to ascertain their function. We employ complementary in vivo zebrafish esrp1/2 mutant assay and in vitro Esrp1/2 murine Py2T cell lines to optimize rigor of approach. We will also discover and functionally validate Esrp-regulated genes, using zebrafish epithelial transgenic reporter lines. We discovered that Esrp1/2 regulates alternative splicing of CTNND1 and will functionally interrogate human CTNND1 gene variants in the zebrafish ctnnd1 mutant in a rescue assay. The expected outcome of this project is to gain mechanistic insights by leveraging genome sequencing data associated with orofacial cleft cohorts, to functionally analyze ESRP1/2 and CTNND1 gene variants in craniofacial development. We will also identify and functionally validate Esrp-regulated genes acting in the periderm. This work will have broader impact by elucidating how regulation of RNA alternative splicing and cell signaling mechanisms are important in periderm and craniofacial morphogenesis.

NIH Research Projects · FY 2026 · 2023-02

Abstract The bone marrow in mammals house both hematopoietic and mesenchymal cells that are responsible for sustaining blood and bone cell production, respectively, throughout adult life. Although the hematopoietic system is well understood, the molecular identities, hierarchy of the marrow mesenchymal cells and their respective contribution to bone homeostasis are just beginning to be unraveled. Elucidation of the organization and functions of the bone marrow mesenchymal cells is fundamental to understanding the pathogenesis of both myeloid and bone diseases. By employing single-cell RNA sequencing (scRNA-seq) technology, we have discovered a subset of bone marrow mesenchymal cells co-expressing adiponectin (Adipoq) and osterix (Osx) which are traditionally considered adipocyte or osteoblast markers, respectively. Trajectory analyses predict the Adipoq+Osx+ bi-marker cells to be common progenitors for osteoblasts and marrow adipogenic lineage cells. Lineage tracing with Osx-CreERT2 or Adipoq-CreERT2 supports that the bi-marker cells give rise to both osteoblasts and adipocytes in vivo. Imaging studies localize the bi-marker cells to the endosteal bone niche. The data therefore support the hypothesis that Adipoq+Osx+ bi-marker cells are adipo-osteoprogenitors attuned to the physiological milieu in the bone marrow. To test the hypothesis, we will first determine the number and fate of the bi-marker cells in young, mature and aged mice to uncover potential age-dependent changes (aim 1). We will then investigate the functional contribution of the bi-marker progenitors to bone formation both under basal conditions and in response to the main bone anabolic drug teriparatide in mature adult mice (aim2). We will finally examine the effect of diabetes on the fate of the bi-marker cells in an experimentally induced type II diabetes mouse model. The studies are expected to shed light on the role of the adipo- osteoprogenitors in bone physiology and pathophysiology.

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract This proposal requests funding for continuation and extension of a phase IIa clinical trial of rifampin, an FDA- approved antibiotic, for safety and efficacy as a treatment for idiopathic infantile hypercalcemia (IIH) due to mutations in the gene encoding CYP24A1 gene. IIH is an uncommon metabolic condition characterized by elevated plasma levels of the activated form of vitamin D, calcitriol, and consequently increased intestinal absorption of calcium and increased bone resorption that together cause hypercalcemia and hypercalciuria. Although IIH typically presents in infancy, patients manifest a life-long defect in vitamin D metabolism that results in hypercalciuria, nephrolithiasis, and renal insufficiency. CYP24A1 encodes the 24-hydroxylase enzyme that represents the principal pathway for inactivation of vitamin D metabolites, and biallelic mutations cause the most common and severe form of IIH. Loss of this pathway allows plasma levels of calcitriol to rise excessively and overcomes feedback mechanisms that should downregulate production of calcitriol. Patients who carry only one defective CYP24A1 allele have a less severe phenotype. There is at present no specific long-term treatment for patients with CYP24A1 mutations and conventional care consists of minimizing sunlight exposure, a low calcium diet, and avoidance of vitamin D-rich foods and vitamin D supplements. This approach does not reduce the risk of renal calcification and renal insufficiency, however, and may lead to low bone density. Thus, there is a significant unmet medical need for safe and effective treatments for this disorder. We have compelling data supporting a therapeutic approach in which the antibiotic rifampin is repurposed to induce expression of CYP3A4, an enzyme that is expressed in the liver and intestine, to provide an alternative pathway for inactivation of vitamin D metabolites. The long-term goal of this project is to develop novel strategies for medical treatment of patients with IIH and other forms of hypercalciuria and nephrolithiasis that are associated with elevated plasma levels of calcitriol. The objective in this application is to determine the optimal safe and effective dose of rifampin that normalizes serum and urine levels of calcium and reduces intestinal absorption of calcium (primary outcomes). Our two complementary goals are to evaluate the extent to which these primary outcomes are related to plasma levels of rifampin, induction of CYP3A4, polymorphisms in the CYP3A4 gene and other genes that influence mineral metabolism, and changes in plasma levels of vitamin D metabolites and to determine the effect of CYP24A1 mutations on bone health. Our central hypothesis is that induction of CYP3A4 by rifampin will reduce levels of calcitriol and thereby decrease intestinal absorption of calcium and we expect that benefits will be related to the extent of CYP3A4 induction. We have access to the necessary study subjects and the expertise and resources to pursue these studies. Our approach is innovative because it proposes to repurpose a well-characterized and safe medication to a new role as a primary therapy for a disorder that currently lacks an effective treatment.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT Obesity increases the risk of developing asthma in children and adults, and obesity-associated asthma (OAA) is often more severe and more difficult to treat than atopic asthma. These poor clinical outcomes may stem from OAA’s distinct immunopathology that includes differences in how lung immune cells respond to inflammatory stimuli and a heterogeneous but neutrophil-predominant lung inflammation. Our limited understanding of how obesity alters lung innate immune cell responses to inflammatory stimuli hinders the development of novel preventative and therapeutic approaches for OAA. Obesity causes lipid deposition in the lung and lipid accumulation in lung tissue resident macrophages (TRMs). These processes may contribute to OAA immunopathology as TRMs are both intimately involved in asthma pathogenesis and sensitive to immunometabolic reprogramming during obesity. Despite this, we do not know if lung TRMs adopt distinct immunometabolic and functional programs as a result of obesity, or if obesity-associated lung TRMs contribute to OAA. To begin to address this knowledge gap, we have performed preliminary studies of lung TRMs in lean and obese mice. We observe lipid-laden TRMs that express proteins characteristic of immunometabolic reprogramming and inflammatory activation in the lungs of obese mice. Using minimally biased lipidomics and in vitro culture techniques, we have identified the fatty acid stearate as a key metabolic signal that may influence lung TRM inflammatory functions during obesity. Finally, we find that obesity and stearate cause activation of the TRM inflammasome—an outcome that is observed in non-lung TRMs during obesity and may contribute to OAA immunopathology. Based on these data, I hypothesize that stearate activates an immunometabolic functional program in lung TRMs that causes exaggerated inflammasome-mediated inflammation in response to innate stimuli. The objectives of this grant are to: (1) identify the lipid signals, cellular metabolic pathways, and inflammatory consequences of obesity-associated lung TRM immunometabolic reprogramming in mice and humans and (2) test the contribution of the TRM inflammasome to OAA-like innate lung inflammation. To attain these objectives, we have developed or obtained novel mouse model systems and established unique collaborations that will allow us to mechanistically interrogate obesity-associated immunometabolic reprogramming of lung TRMs in mice, and translate our observations to pediatric and adult subjects. Doing so will identify molecules and pathways that can be targeted by future OAA-specific therapeutics, and inform studies of other obesity-associated inflammatory lung diseases.

NIH Research Projects · FY 2026 · 2022-11

Kingella kingae is an invasive gram-negative pathogen that has been recognized recently as a leading cause of bone and joint infections in young children, accounting for up to 88% of osteoarticular cases in children <4 years old. In addition, K. kingae is an important cause of invasive bloodstream infections in young children. Complications of osteoarticular infections in children include abnormalities in bone growth, limitation of joint mobility, unstable joint articulation, and chronic joint dislocation, resulting in residual skeletal dysfunction in 10- 25% of cases. Complications of invasive bloodstream infections include multi-organ injury and mortality. Approximately 25% of K. kingae isolates possess β-lactamase activity, and many of these isolates are resistant to other antibiotics as well, raising concern about approaches to treatment in the future. At present there are no effective strategies to prevent K. kingae disease and the associated morbidity. The pathogenesis of K. kingae disease begins with colonization of the oropharynx, followed by invasion of the bloodstream and spread to bones, joints, and other sites. We have established that isolates of K. kingae produce an exopolysaccharide that is encoded by the pamABCDE locus, is a homopolymer of galactofuranose, is secreted from the organism, and is a critical virulence factor essential for full virulence. We have found that there are 2 distinct exopolysaccharide structures, distinguished by the linkage of the galactofuranose repeating subunit and referred to as type 1 and type 2. Importantly, the exopolysaccharide promotes resistance to serum- mediated killing and neutrophil phagocytosis and thereby promotes K. kingae survival in the bloodstream, indicating that at least some of the exopolysaccharide is anchored to the bacterial surface. Preliminary results indicate that pooled serum from healthy adults and convalescent serum samples from children with invasive K. kingae disease contain antibodies against the exopolysaccharide. In this proposal, we will elucidate the mechanism by which the type 1 and type 2 exopolysaccharides are synthesized and anchored to the bacterial surface. In addition, we will elucidate the pathogenic properties of the type 1 and type 2 exopolysaccharides. We will also elucidate the immunogenicity and protective efficacy of the exopolysaccharides. The proposed studies will provide fundamental insight into K. kingae pathogenicity and basic aspects of bacterial exopolysaccharides. These studies will also facilitate development of a K. kingae vaccine and antibody-based therapeutics against other pathogens with galactofuranose-containing surface structures.

NIH Research Projects · FY 2025 · 2022-09

Mild traumatic brain injury (mTBI) is a common injury in childhood with the potential to have a substantial impact on function in the school and sports setting. Early recognition, followed by up-to-date mTBI care, has been demonstrated to improve outcomes, reducing the incidence of persistent post-concussive symptoms (PPCS). The current best evidence for mTBI care has been brought together in the CDC’s Pediatric mTBI Guideline, poised to be translated into clinical care via rigorous, planned implementation. The Minds Matter Concussion Program at the Children’s Hospital of Philadelphia (CHOP) has demonstrated a track record of positively impacting the care of children with mTBI via the implementation of electronic clinical decision support (eCDS) embedded in the network-wide electronic health record. The goal of this project is to build on that experience to improve the outcomes of children with mTBI and reduce the disparities in those outcomes by translating the current Guideline into standard care. The first aim will be directed at understanding the specific roots of the current disparities in pediatric mTBI outcomes for over 8000 children by examining the Minds Matter Concussion Registry, a census of children seen for mTBI across the CHOP healthcare network. In addition, we will evaluate school outcomes through our partnership with BrainSTEPS, a program administered by the Brain Injury Association of Pennsylvania. In the second aim, through our ongoing successful partnership with the CHOP Primary Care Network, we will implement Guideline-informed eCDS utilizing a validated clinical prediction score (5P risk score) and targeted mTBI physical examination (visio-vestibular examination) in selected urban and suburban primary care practices in a stepped-wedge trial. This effort will quantify the direct impact of the Guideline on improving pediatric mTBI outcomes, including the identification of children at high risk of PPCS in need of referral for specialized care, and by including diverse practices, the effectiveness of the intervention in reducing disparities. As part of the trial design, an urban and suburban primary care practice will also participate in the novel implementation of a mobile health (mHealth) intervention using a smartphone application to perform ecological momentary assessment of symptoms to identify those in the moderate risk category who would benefit from earlier referral. The third aim will investigate and address barriers to accessing specialized mTBI care from alternative points of entry into the healthcare system, namely urban schools and youth club sports organizations by conducting qualitative semi-structured interviews and providing mTBI education to those personnel based on the Guideline. We will implement an innovative telehealth athletic training consultation service line and examine its impact on clinical outcomes and reducing disparities. This project, when completed, in addition to generating new knowledge about outcomes and disparities in pediatric mTBI, will also produce the deliverables of eCDS for clinical implementation of the Guideline, including novel mHealth and telehealth approaches to improving outcomes and reducing disparities in mTBI, impacting the lives of children broadly.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract Dr. Irit R. Rasooly’s goal is to become an independent clinician scientist dedicated to promoting accurate, timely, and equitable pediatric diagnosis. To further develop the novel skillset necessary to apply data-driven, systems-informed approaches to improve diagnosis, Dr. Rasooly’s career development plan incorporates mentored research and training in clinical informatics, longitudinal analysis, human factors engineering, implementation science, and trauma informed care. She will be mentored by experienced investigators with whom she has established, productive collaborative relationships: Dr. Christopher P. Bonafide (Primary Mentor) is an expert in developing, evaluating, and implementing interventions at the intersection of patient safety and technological innovation. Dr. Joanne N. Wood (Co-Mentor) is an expert in child physical abuse research whose scholarship has focused on racial disparities and variation in abuse evaluations. Dr. Kathy Shaw (Co-Mentor) is a national and intuitional leader in patient safety science and diagnostic error. The Children’s Hospital of Philadelphia and University of Pennsylvania are deeply supportive of Dr. Rasooly’s work and provide an unparalleled research training environment. Missed diagnosis of child abuse, in which abusive injury goes unrecognized or is misattributed to accidental trauma, is among the gravest examples of pediatric diagnostic error and a serious threat to pediatric health. While electronic health record (EHR) clinical decision support (CDS) has shown promise in improving timeliness and reducing disparities in abuse diagnosis in emergency department settings, there is a critical need for strategies to support diagnosis in primary care, where most pediatric care is delivered and where there may be opportunity to intervene before children sustain serious injuries. To lay the groundwork for creation of CDS to improve abuse diagnosis in primary care, the objective of this work is to apply EHR data- and systems-analysis to inform identification of strategies to support diagnosis of child physical abuse in primary care. Specific aims are to: (1) Detect and validate markers of physical abuse in longitudinal, clustered, EHR-derived data to distinguish young children experiencing abuse from matched controls, (2) Determine causes of abuse-related diagnostic error in primary care by conducting scenario-based high-fidelity human factors engineering EHR simulations, and (3) Identify and prioritize feasible, acceptable, and appropriate CDS strategies through a consensus approach. Findings will inform development of CDS to be tested in a future multi-center, R01 funded clinical trial. Completion of the proposed research and training will position Dr. Rasooly to launch an independent research career advancing pediatric diagnostic excellence and reducing health disparities in primary care, thereby addressing multiple AHRQ research priority areas.

NIH Research Projects · FY 2024 · 2022-09

Project Summary/Abstract Properties of disease transmission can evolve throughout the pandemic and may be influenced by health policy decisions, regional demographic characteristics, community behaviors, environmental characteristics, and population immunity. This proposal is motivated by significant challenges we have encountered, including dynamic connections between virus evolution, health policy, population behavior and degree of immunity over time, and evolving data elements and data quality due to varying testing criteria and inconsistent reporting behavior. The overarching goal of this proposal is to develop a framework of pandemic predictive intelligence that can adapt over time to changing data quality and evolving behavioral and environmental characteristics that influence disease transmission. The key advantage of the proposed modeling approach is its adaptation to time varying exposures of community behavior and mobility, environmental conditions, mitigation strategies, population immunity, and viral evolution. Through three projects, we will develop models to 1) improve the forecasting accuracy by enhancing model robustness (robustness to data error and model assumptions), 2) connect the dots between viral evolution and transmissibility, and 3) advance the state-of-the-art forecasting by integrating five major components, including viral evolution, transmissibility, social behavior, population immunity and public health policy, to build a learning system for predictive modeling for infectious disease. To ensure the broader impact of the proposed research, we will develop, validate, and evaluate methodology and software for pandemic forecasting, real-time monitoring, mitigation, and prevention of the spread of pathogens using national county/city-level data from the US Department of Health and Human Services, the University of Pennsylvania, and other publicly available data resources. The proposed work will contribute to foundational work needed to advance pandemic science, which includes predictive modelling of pandemic and evidence-assisted health policymaking for pandemic prevention and response.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY/ABSTRACT The purpose of this Career Development Award is to prepare Dr. Maria Kate Henry, MD, MSCE for her long- term goal to serve as an independent clinical investigator developing evidence-based clinical decision support that optimizes the equitable use of imaging to inform the identification, care, and protection of victims of child abuse. Infants with identified injuries that raise concerns regarding potential abuse (e.g., femur fracture) can also have brain injuries that are clinically occult and only detectable by neuroimaging (computed tomography, magnetic resonance imaging). Detection of a clinically occult head injury (e.g., subdural hematoma) guides medical care, dramatically increases the level of diagnostic certainty regarding abuse, and informs child protection decision making. Neuroimaging, however, is not without risk, forcing clinicians to weigh the risks of missing an intracranial injury with the risks of radiation or sedation. While clinicians have clear guidance on use of plain radiography to detect occult fractures when abuse is suspected, they lack clear guidance on use of neuroimaging, contributing to variation and disparities in care. In the proposed study, Dr. Henry will develop the Child Abuse Pediatrics Risk of Intra-Cranial Injury (CAPRICI) clinical decision rule (CDR) that identifies which well-appearing infants with concern for abuse warrant neuroimaging. Specifically in Aim 1, Dr. Henry will use a machine learning approach to adapt and improve a predictive model quantifying risk of intracranial injury among infants undergoing evaluations for abuse in CAPNET, a multicenter child abuse research network. In Aim 2, Dr. Henry will use the RAND/UCLA Appropriateness Method to determine the thresholds at which screening for intracranial injury is warranted based on the probability of injury identification. Based on the findings from Aims 1 and 2, Dr. Henry will create a clinically-informed CAPRICI CDR. This proposal and Dr. Henry’s long-term goals align squarely with AHRQ’s mission of producing evidence to make health care safer, higher quality, and equitable, as well as AHRQ’s research priority of “harnessing data and technology to improve health care quality” in an AHRQ priority population (children). Towards Dr. Henry’s goal of becoming an independent clinical investigator, she and her mentors have developed a comprehensive 5-year career development plan that integrates with the research proposal to ensure that Dr. Henry develops skills in (1) rigorous analysis of multicenter data, (2) CDR development and implementation, (3) modified consensus methodology, and (4) professional development (grant writing, mentorship, presentation, and leadership skills). This plan includes tailored mentorship from local and national leaders, formal didactics, professional development, and an innovative research approach. At the conclusion of this award, Dr. Henry will be well- positioned to apply for a multicenter R01 assessing the impact of implementation of the CDR on clinical practice in CAPNET.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Sepsis, defined as a dysregulated host response to infection, is responsible for 75,000 childhood hospitalizations annually in the United States and 50% of inpatient pediatric deaths worldwide. Despite multiple trials, there are no targeted therapies for adult or pediatric sepsis, and supportive care with timely antibiotics and fluid resuscitation remains the mainstay of therapy. Fluid therapy is a cornerstone of sepsis resuscitation, although the choice of fluid remains controversial. Recent trials in adults showed lower rates of major adverse kidney events within 30 days (MAKE30) with balanced crystalloid, such as lactated Ringer’s (LR), relative to normal saline (NS). MAKE30 is a composite endpoint incorporating persistent kidney dysfunction, initiation of dialysis, or death. Balanced solutions like LR have a composition more similar to normal human serum, whereas NS has been associated with worse kidney function, albeit via unclear mechanisms. While much of pediatric sepsis management is extrapolated from adults, children have a distinct epidemiology and outcome profile, making application of adult data problematic. Thus, to specifically assess whether LR or NS resuscitation improves MAKE30 in children, the PRagMatic Pediatric Trial of Balanced versus nOrmaL Saline FlUid in Sepsis (PRoMPT BOLUS) was undertaken, with planned enrollment of 8,800 children. However, heterogeneity has contributed to negative trials in sepsis, as therapies effective in some patients are ineffective in others. To mitigate this heterogeneity, biomarkers have been proposed for both risk stratification and identification of sub-phenotypes with shared pathophysiology. Our study, entitled Finding Appropriate Subtypes in a Trial of Balanced versus nOrmaL Saline FlUid in Sepsis (FAST BOLUS), will assess for heterogeneity of treatment effect of LR versus NS in the PRoMPT BOLUS trial. Leveraging our group’s extensive experience with biomarker-defined risk stratification and sub-phenotyping, we will measure plasma biomarkers in 800 children collected at randomization and assess the for differential effects of fluid assignment on MAKE30 across risk strata using two separate biomarker-based mortality prediction models (Aim 1). Additionally, we will test for differential effects of fluid assignment on MAKE30 after stratifying subjects according to one of two biomarker-defined inflammatory sub-phenotypes (Aim 2). Lastly, we will leverage this biobank to investigate potential mechanisms between fluid choice and MAKE30 by measuring markers of endothelial cell (angiopoietin-2) and endothelial glycocalyx (syndecan-1) damage (Aim 3). These Aims will assess the utility of established biomarker-based strategies for prognostic (Aim 1) and predictive enrichment (Aim 2) strategies by leveraging an ongoing pediatric sepsis trial, which is a necessary analysis of randomized trials conducted in heterogeneous critical illness syndromes. Successful completion of the Aims will provide evidence for personalized fluid resuscitation in pediatric sepsis and establish proof of concept data for incorporation of biomarker-based methods to reduce heterogeneity in future trials.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY / ABSTRACT Detailed information about the state of a cell, e.g., its lineage, mitotic history, proliferation potential, and functional competence, consolidates through epigenetic modifications of its DNA and chromatin. Among these modifications, DNA methylation has been widely studied and profiled to dissect tissue heterogeneity, disease cell of origin, and implement liquid biopsy-based disease diagnosis, thanks to its chemical stability and genome- wide distribution. Compared to bulk tissue methylome assays, which yield convoluted, hard-to-decipher signals from thousands to millions of cells, single-cell DNA methylome profiling is advantageous in cell identity-related applications. Despite the rapid increase in the volume and variety of single-cell DNA methylome data in recent years, availability of powerful and easy-to-use computational tools for their analyses is still an unmet demand. Consensus on the optimal strategy of interpreting cell states based on single-cell methylome data has not been reached. My lab’s long-term goal is to elucidate epigenetic cell identities at the single-cell level in humans and mice. Towards that goal, I propose to develop a suite of computational tools, for analyzing single-cell methylation data, that will encompass functions for data preprocessing, quality control, imputation, methylome signature extraction, cell state annotation, and exploratory visualization. These software tools will be engineered to be efficient, modular, and will be designed to operate both in high-performance computing environments and on basic laptops. These tools would be able to alert the investigator of potential data quality issues, feedback to accelerate methylation assay development, and discover biological links between the DNA methylome and the cell’s genetic makeup, mitotic history, cell-cycle stage, differentiation capacity, and functional state. They can also be used to study cell population traits in bulk tissue samples. Together with these computational tools, we also aim to distribute a cell-type-resolution reference methylome catalog to benefit the research community. My proposed work will deliver computational tools and methylation references to deepen our understanding of the role of DNA methylation in determining cell lineages and provide practical tools for epigenetic cell typing. The methods to be developed could be readily plugged into exploratory and translational applications in broader biomedical contexts.

NIH Research Projects · FY 2025 · 2022-09

During the summer of 2021, 80% of pediatricians reported using live audio-video telemedicine in the prior month, up from 16% of pediatricians with telemedicine experience just four years before. The child health workforce now actively uses telemedicine to care for children, but lacks evidence to guide best practices for telemedicine use in pediatric primary care. Our research team previously found gaps in the quality of care delivered by commercial direct-to-consumer telemedicine for the most common acute pediatric concerns (acute respiratory tract infections). We also demonstrated improvement in management of common chronic conditions (e.g., asthma, attention-deficit/hyperactivity disorder) using telemedicine in research settings. However, the child health community lacks data regarding the use of telemedicine in evolving “real-world” primary care practices, including how telemedicine visits compare to in- person visits for the same conditions and how telemedicine use impacts longitudinal outcomes for primary care patients. Additionally, we lack understanding of the structures and processes that support primary care practices in using telemedicine in ways that optimally enhance child health. The overarching goal of this proposal is to identify actionable strategies for promoting the use of telemedicine within primary care in ways that improve child health, using data from primary care practices from 2018-25. In Aim 1, we will compare visit-level quality of primary care visits for children delivered through telemedicine vs. in-person using electronic health record data from over 1,000 practices including independent practices, health system-affiliated practices, and community health centers. In Aim 2, we will compare child health outcomes for children receiving care in primary care practices with higher vs. lower telemedicine use through comparative interrupted time series analysis of metrics related to preventive, acute, and chronic care. Finally, in Aim 3, we will identify the practice structures and processes that promote or impede use of telemedicine in primary care in ways that improve child health through a qualitative multiple-case study. Through this rigorous mixed-methods approach informed by the Systems Engineering Initiative for Patient Safety (SEIPS) 2.0 model, we will identify visit-level and child-level impact of telemedicine integration within primary care for children as well as processes supporting health-promoting use of telemedicine. As a result, this examination of telemedicine across preventive, acute, and chronic care activities overall, for subpopulations, and within exemplar practices will provide critical knowledge to inform research, practice, and policy to optimize ongoing use of telemedicine within primary care to promote child health.

NIH Research Projects · FY 2026 · 2022-09

Technical Abstract It is now appreciated that the adaptive immune system plays an integral role in the removal of toxic Aβ oligomers from Alzheimer disease (AD) brains. This is accomplished through T effector cell entry into the brain via the choroid plexus (CP) and their subsequent interaction with resident microglia. However, the role of the T regulatory (Tregs) cells, which moderate Teff antigen inflammatory response, in the removal of Aβ oligomers is currently debated. We have found that Treg cells are oxidative while Teff cells are glycolytic and that AD is associated with chronic mitochondrial oxidative phosphorylation (OXPHS) defects and increased mitochondrial reactive oxygen species (mROS) production. Therefore, we hypothesize that preexisting differences in mitochondrial OXPHOS and mROS production can predispose to Alzheimer disease amyloid accumulation and cognitive decline due to chronic neuronal cell damage, stimulation of toxic Aβ oligomer formation, and alteration in the Treg control of immune function. To test this hypothesis, we propose three specific aims. First, to determine the importance of mitochondrial defects in AD, we will combine the classical nuclear DNA (nDNA) APPswe AD transgene with mtDNAs harboring defined OXPHOS defects (COIV421A and ND6P25L) or established Treg suppressive function (mtDNAB6 and mtDNANZB) and document their effects on clearance of Aβ plaques and restoration of cognitive function. Second, to determine the role of Treg in modulating Aβ pathology and cognition, we will use adoptive transfer of weakly Teff-suppressive mtDNANZB Treg cells and strongly Teff-suppressive mtDNAB6 Treg cells in APPswe mtDNAB6 or mtDNANZB mice and evaluate Aβ plaque removal and cognitive function. Lastly, to determine the effects of mitochondrial OXPHOS defects and mROS production on the central nervous system, CP, Treg cells, and microglia in terms of Aβ pathology and cognitive decline, we will use tissue-specific Cre recombinases to either 1) inactivate the nuclear Ant2fl gene thereby reducing OXPHOS or 2) activate the mitochondrially-targeted anti-oxidant mCATfl gene, thereby reducing mROS in Tg2576 APPswe + mtDNAB6 or mtDNANZB mice. According to our hypothesis, the first specific aim is predicted to confirm that mitochondrial dysfunction is causally related to AD and that mitochondria modulate the anti-Aβ oligomer removal by Treg cells. The second specific aim is predicted to confirm the importance of Treg mitochondrial function in Aβ plaque removal. The last specific aim is predicted to confirm the role of partial OXPHOS defects in the brain and microglia in predisposition to AD and to establish the importance of Treg mROS in modulating Aβ plaque removal and cognitive pathology. Should these predictions be born out, they may suggest that therapies to enhance mitochondrial function and reduce mROS may prove beneficial in treating AD.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Brain electrophysiological phenotyping holds promise for parsing heterogeneity in ASD and enabling testing of proposed treatments more reliably in biologically-based subgroups of ASD. There have been considerable advances in the non-invasive imaging and electrophysiologic correlates of phenotypes in ASD, in children, including 1) delayed auditory evoked response components (M50, M100 latency); 2) delayed magnetic mismatch fields (MMF) elicited by vowel contrasts as well as atypical rightward lateralization; 3) atypical development of gamma-band oscillatory phase synchrony, coupled to atypical levels of inhibitory neurotransmitter, GABA; and 4) atypical motor oscillatory activity, particularly the post-movement beta rebound (PMBR). We have suggested that such electrophysiological signatures might serve as biomarkers in stratifying patients for inclusion in clinical trials according to biology, rather than behavior alone. However, little is known about how these candidate biomarkers mature into adulthood. This is important because there are millions of adults on the autism spectrum, presenting in clinic in need of treatment. Our preliminary studies suggest that there might be persistence of childhood electrophysiological phenotypes into adulthood (in particular, auditory M50/M100 and MMF delays), while differences in auditory gamma band phase synchrony and motor PMBR oscillatory responses may in fact emerge during adolescence. If indeed childhood biological differences persist into adulthood and/or some biological differences emerge in late adolescence/ early adulthood, then the opportunity and target for remediation may also persist in adulthood. This would indicate the need for a quantitative biomarker for both stratification (inclusion/exclusion) purposes but also for monitoring treatment target engagement, as well as longer term evidence of brain response. We will recruit 72 autistic adolescents/ adults (14-45yrs, 48M, 24F) and 72 age-/sex-matched typically developing (TD) peers into a multimodal imaging study with a 12-week longitudinal design to mimic a typical pharmaceutical trial and establish precision estimates for each metric to define the resolution of interval change in subsequent trials. We will carry out the following Aims. In Aim 1, we will evaluate, in a sample of adults, the group level ASD vs TD discrimination of each of a battery of MEG metrics and assess intra- and inter-subject variability over three scan sessions (baseline, 4weeks, 12weeks). This will establish the effect size required of any putative pharmaceutical. In Aim 2, we will use multimodal imaging to address heterogeneity and probe the biological underpinnings of M50 latency prolongation in adults. In Aim 3, we will use simultaneously-acquired MEG and EEG, to determine the efficacy of EEG analogs of the proposed MEG measures to achieve similar group- level discrimination of individuals with ASD vs TD. EEG is lower-cost, simpler to perform and has widespread availability appropriate for clinical trial conduct. In culmination, the aims of this study will provide pivotal answers to critical “clinical readiness” questions about electrophysiological biomarkers in autistic adults.

NIH Research Projects · FY 2024 · 2022-09

Project Summary Brain electrophysiological phenotyping holds promise for parsing heterogeneity in ASD and enabling testing of proposed treatments more reliably in biologically-based subgroups of ASD. There have been considerable advances in the non-invasive imaging and electrophysiologic correlates of phenotypes in ASD, in children, including 1) delayed auditory evoked response components (M50, M100 latency); 2) delayed magnetic mismatch fields (MMF) elicited by vowel contrasts as well as atypical rightward lateralization; 3) atypical development of gamma-band oscillatory phase synchrony, coupled to atypical levels of inhibitory neurotransmitter, GABA; and 4) atypical motor oscillatory activity, particularly the post-movement beta rebound (PMBR). We have suggested that such electrophysiological signatures might serve as biomarkers in stratifying patients for inclusion in clinical trials according to biology, rather than behavior alone. However, little is known about how these candidate biomarkers mature into adulthood. This is important because there are millions of adults on the autism spectrum, presenting in clinic in need of treatment. Our preliminary studies suggest that there might be persistence of childhood electrophysiological phenotypes into adulthood (in particular, auditory M50/M100 and MMF delays), while differences in auditory gamma band phase synchrony and motor PMBR oscillatory responses may in fact emerge during adolescence. If indeed childhood biological differences persist into adulthood and/or some biological differences emerge in late adolescence/ early adulthood, then the opportunity and target for remediation may also persist in adulthood. This would indicate the need for a quantitative biomarker for both stratification (inclusion/exclusion) purposes but also for monitoring treatment target engagement, as well as longer term evidence of brain response. We will recruit 72 autistic adolescents/ adults (14-45yrs, 48M, 24F) and 72 age-/sex-matched typically developing (TD) peers into a multimodal imaging study with a 12-week longitudinal design to mimic a typical pharmaceutical trial and establish precision estimates for each metric to define the resolution of interval change in subsequent trials. We will carry out the following Aims. In Aim 1, we will evaluate, in a sample of adults, the group level ASD vs TD discrimination of each of a battery of MEG metrics and assess intra- and inter-subject variability over three scan sessions (baseline, 4weeks, 12weeks). This will establish the effect size required of any putative pharmaceutical. In Aim 2, we will use multimodal imaging to address heterogeneity and probe the biological underpinnings of M50 latency prolongation in adults. In Aim 3, we will use simultaneously-acquired MEG and EEG, to determine the efficacy of EEG analogs of the proposed MEG measures to achieve similar group- level discrimination of individuals with ASD vs TD. EEG is lower-cost, simpler to perform and has widespread availability appropriate for clinical trial conduct. In culmination, the aims of this study will provide pivotal answers to critical “clinical readiness” questions about electrophysiological biomarkers in autistic adults.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The pulmonary mesenchyme, which includes fibroblasts and smooth muscle, provides critical support for resident epithelial progenitors, but how it contributes to lung injury resolution is unclear. As a barrier organ the lung is constantly exposed to inhaled particulates, allergens, and respiratory pathogens. This barrage of exposures is countered by mucous secretions, antimicrobial agents, and sentinel immune cells. In the case of a more severe insult that causes lung damage, the inflammatory response and activation of wound-healing fibroblasts ensues. The research program described here seeks to elucidate how signaling between resident fibroblasts and immune cells controls the length and successful outcome of the lung injury repair response. Further, this research will test a new model proposed for the origins of human lung disease by testing how dysfunctional epithelial cells perpetuate the inflammation-driven fibroblast response. The overarching goal of this program is to determine the molecular pathways utilized by distinct fibroblast subsets and how they converge to shape the immune status of the lung. These questions will be explored by using our unique genetic mouse models for fibroblast lineages and in vivo models of respiratory dysfunction including chemical-induced injury or influenza infection. Moreover, we propose to employ a combination of sequencing analysis and establish novel organoid models to explore the interactions of fibroblast subsets and tissue resident immune cells. Together, our efforts will provide new insights in the mechanisms of lung repair resolution and build a foundation for novel future contributions.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT The cell-to-cell connections that comprise the functional circuitry and global network organization of the human brain have at their root precise spatiotemporal patterns of gene expression that differentiate brain regions, cell types, and developmental periods. Recent discoveries have revealed highly dynamic spatiotemporal patterns of gene expression throughout development and in several regions of the brain, but elucidating the neural connectome – the comprehensive map of all neural connections – at multiple ages of human development has, until recently, not been practical. Consequently, although we now have a strong understanding of transcriptional reorganizations in the developing and adult human brain, how these dynamics mediate other key phenomena essential for brain function, such as neural connectivity, is not well understood. As region-specific disruptions to neural connectivity may underlie neuropsychiatric disorders including schizophrenia and autism spectrum disorder, the association of transcriptome information with connectivity is a critical next frontier for analysis. We therefore propose to integrate human brain connectome based on neural MRI and transcriptome data collected across multiple timepoints in human postnatal development with unprecedented spatial resolution to implicate candidate genes, gene expression modules, brain regions, developmental periods, and connectivity patterns with human brain development. To accomplish this, we will develop a high-resolution connectome atlas established with unique existing developmental and adult data as well as next-generation connectome data from early and late childhood being acquired with other sources. This atlas will be paired with transcriptomic atlas from the postnatal human neocortex consisting of 25 neocortical areas and will be integrated with publicly available, less comprehensive transcriptomic data generated at the level of the single cell. Finally, we will validate gene expression patterns and the associations we reveal, and freely disseminate atlases and datasets through an integrated web portal.