Cincinnati Childrens Hosp Med Ctr

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT This proposal for the K01 Mentored Research Scientist Career Development Award is intended to support my long-term goal of becoming a trailblazing independent investigator dedicated to transforming outcomes and dismantling inequities in youth violence. To reach this goal, I seek to gain the pivotal knowledge and skills to design and rigorously evaluate community-based interventions to reduce youth violence. Firearm injury is the leading cause of death for children and adolescents in the United States, with the burden of these injuries disproportionately born by disadvantaged communities. In this proposal, I seek to address the research priority of youth violence, and specifically firearm injuries, using complex systems science and human-centered design approaches. My central hypothesis is that these tools will allow us to design feasible, acceptable, and effective interventions to reduce youth gun violence. I will pursue the proposed research and career development goals with the following aims: Aim 1. Identify areas of greatest impact for reducing youth firearm violence in Cincinnati using system dynamics modeling. I will use system dynamics modeling and our existing co-produced qualitative map to identify areas within our complex dynamic system where interventions may have the greatest impact on reducing rates of youth firearm violence. I hypothesize that we will identify 1-2 areas ripe for intervention. Aim 2. Co-design an intervention to reduce youth firearm violence in one neighborhood in Cincinnati. I will utilize human-centered design strategies to develop at least one intervention expected to reduce youth violence. I hypothesize that an inclusive community-centered, multidisciplinary team can design a feasible, acceptable, effective intervention targeting upstream structural determinants of youth violence. My training goals for this proposed award are to gain experience in (1) complex systems science methods, (2) human-centered design, and (3) community-engaged participatory research. I seek to position my research and skills at the intersection of these three methodologic approaches. I will use findings and skills gained from this K01 proposal to develop a competitive R21, and subsequently R01, proposal on testing and implementation of the designed intervention. I will complete this proposed work as an Assistant Professor of Surgery and Attending Pediatric Surgeon at Cincinnati Children’s Hospital Medical Center, with significant and enthusiastic support from my divisional and departmental leadership, as well as my proposal mentors.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY: Atrial fibrillation (AF) is the most common sustained arrhythmia in the United States at 5% of the population and 1-in-3 lifetime risk. Treatment and care associated with AF costs over $26 billion/year in the US and accounts for 10% of all Medicare spending. Debilitating complications linked to AF include heart failure and stroke. The molecular links between genetics and AF disease risk are not well understood and pose a significant knowledge gap for curative therapies and patient care. Common coding and regulatory variations of the transcription factor (TF), PITX2, have been resoundingly linked to AF risk. PITX2 is known for regulating left-right asymmetry during development. In the heart, PITX2 is expressed in the left atrial (LA) and pulmonary vein (PV) myocardium, the most common trigger sites for AF. Mouse models of reduced Pitx2 are susceptible to AF, and my recent work describes how decreased Pitx2 perturbs TF networks and hints at a developmental origin of AF risk; however, the mechanism is unknown. The central hypothesis for this study is that PITX2 drives left-sided fate determination through epigenetic remodeling, which imparts long-lasting transcriptional consequences underlying adult disease. To address this hypothesis: 1. Investigate the Pitx2-dependent DNA methylome during left-right determination, analyzing observed DNA methylation pattern differences at PITX2 binding sites. 2. Examine the role of PITX2 in oxidative stress response and homeostasis, considering the unique oxygen-rich environment of the PV and preliminary findings of PITX2 interactions with factors associated with the DNA damage response. 3. Elucidate the epigenetic mechanisms of reversible PITX2-mediated transcriptional repression, focusing on PRMT5 mediated-histone modifications. Altogether, the proposed project aims to understand the role of PITX2 in epigenetic patterning of the LA in context of AF. The proposed study takes a novel approach to interrogating the PITX2 gene regulatory network and develops several new reagents and datasets for the field. Furthermore, through this work, I hope to gain insights for more targeted approaches in AF patient care.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Hematopoietic stem cells (HSCs) can self-renew and generate all mature blood cells, allowing them to be used for transplantation to treat hematological disorders. However, as many as 5-27% of patients experience poor graft function, or incomplete hematopoietic recovery after transplant, which has only 6% 2-year overall survival. This can be a direct result of defective HSC self-renewal and multilineage differentiation potential. However, the specific pathways that regulate HSC function remain poorly understood. HSCs are mostly quiescent with low metabolic and mitochondrial activity. They are activated into cell cycle and their mitochondrial activity increases following transplant or 5-fluorouracil (5-FU)-induced myeloablation. Prior work in our lab has shown that after replicative stress, HSCs have decreased regenerative ability, dysfunctional mitochondrial dynamics, and decreased expression of metabolic genes such as stearoyl-Coa desaturase 1 (Scd1). SCD1 is an enzyme that catalyzes the formation of monounsaturated fatty acids (MUFAs) in the de novo lipid synthesis pathway. These products are used to form triglycerides and phospholipids that alter cell membrane structure, signaling, and mitochondrial homeostasis. Specifically, lipids are necessary for proper mitochondrial membrane architecture, dynamics, and oxidative metabolism. Our preliminary data demonstrate that pharmacological inhibition of SCD1 in HSCs in vitro significantly decreases their regenerative ability. SCD1 inhibition also leads to changes in mitochondrial network structure. However, the contribution of MUFA synthesis to HSC function in vivo and why HSCs depend on this pathway is unknown. Our central hypothesis is that MUFA synthesis is important for maintaining HSC regenerative function by supporting mitochondrial function, and loss of this pathway after replicative stress causes HSC functional decline. We will address this hypothesis by determining the role of MUFA synthesis in HSC function in vivo (Aim 1) using an inducible SCD1 knockout mouse model. We will then assess HSC regenerative function by serial competitive transplantation and recovery after 5-FU-induced myeloablation. We will also determine the mechanism by which MUFA synthesis maintains HSC function (Aim 2) by assessing mitochondrial function and performing lipidomic and metabolomic analysis in SCD1-deficient conditions. Using oleic acid, a product of the SCD1-catalyzed reaction, to restore MUFAs, we will determine whether this improves HSC regenerative and mitochondrial function after replicative stress. These studies will lead to a global understanding of the importance of MUFA synthesis to HSC function and reveal pathways that can be targeted to improve HSC regenerative functions and clinical outcomes of HSC-based therapies.

NIH Research Projects · FY 2025 · 2025-09

Abstract Pediatric-onset chronic musculoskeletal pain disorders are common, but the mechanisms underlying their pathophysiology are poorly understood. Current best practices in pediatric pain management focus on symptom management and restoration of function. Absent a more mechanistic understanding of the drivers of pediatric chronic pain and potential sub-types of chronic pain based on underlying pathophysiology (as opposed to phenotypic classifications), pain providers are unable to target treatments to the root causes of patients’ symptoms. There is no cure for pediatric-onset chronic pain, and many children and adolescents continue to have pain in adulthood, highlighting the need for greater research into the development of novel treatment strategies. At Cincinnati Children’s Hospital Medical Center, we have a large and well-established multidisciplinary team of investigators who are already working to unravel the complex biopsychosocial contributors to chronic musculoskeletal (MSK) pain in children. Our multiple ongoing registries and cohorts have more than 2400 patients with pediatric-onset primary MSK pain disorders, pediatric rheumatic diseases, and surgical interventions for whom we have rich medical history, pain-related and psychosocial data. The proposed research would leverage these existing registries and cohorts to enroll a heterogeneous sample of N=600 adolescents and young adults (ages 16-26); N= 400 with primary chronic MSK pain, or chronic MSK pain secondary to rheumatic disease or major MSK surgery, and N=200 with a rheumatic disease or history of major MSK surgery, but no chronic pain. Using a mixed retrospective-prospective cohort study design, we first propose to combine and harmonize retrospective data from patients’ past clinical registry and research records. We then will acquire prospective data regarding pain characteristics, quantitative sensory testing and cytokine profiling and apply a model-based unsupervised clustering analytic approach to define mechanistically distinct chronic MSK pain subgroups. Further, we will use serum samples from patients to conduct in vitro experiments to identify neuro-immune signaling pathways impacting sensory transduction using an innovative induced Pluripotent Stem Cell (iPSC) model. This mechanistic examination will be further augmented by analysis of how psychosocial, behavioral and lifestyle factors mediate pain-related outcomes among participants. In pursuing an analysis of patients that extends from the level of community characteristics to single-cell assays, this research will result in the construction of a robust biopsychosocial model in which patients across multiple pain sub-groups may be classified according to genetic and cellular mechanisms as well as psychological, behavioral, and social features promoting risk and resilience. Through collaboration across disciplines along with patient advisory partners, this study will reveal mechanistically driven treatment targets for future therapeutic trials as well as novel insights into the relationship between painful MSK conditions of childhood and the biopsychosocial factors that contribute to patient outcomes.

NIH Research Projects · FY 2025 · 2025-09

Asthma affects over 10 million youth in the United States and is associated with substantial morbidity, emotional impact, and an annual $56 billion in costs of care. One major contributor to the significant public health burden of asthma is nonadherence to prescribed medical treatment, with rates of non-adherence up to 70% in adolescents with asthma. Our work has demonstrated that although behavioral interventions improve adherence generally, interventions are not effective for all children with asthma. Our iteratively developed stepped care mHealth intervention (Asthma Ctrl) was designed to improve adherence by using an adaptable intervention conserving patient, family, and provider time, costs, and resources. Results from our pilot RCT demonstrated feasibility, acceptability and clinically significant improvements in adherence and asthma outcomes. The aim of this proposed multi-site R01 is to conduct a two-stage, sequential, multiple assignment, randomized trial (SMART) to evaluate the effectiveness of our stepped-care mHealth intervention strategies for improving adherence to daily inhaled corticosteroids in adolescents with asthma for adolescents with low adherence or key health drivers related to adherence. A one-month washout and one-month baseline period will be followed by two stages. In Stage 1 (1 month), patients will be randomized to receive an mHealth asthma management app providing education and automated digital medication reminders (control) or individualized adherence feedback via text messaging based on real-time adherence monitoring (Asthma Ctrl; treatment). At the beginning of Stage 2 (two months), patients randomized to treatment who do not achieve adherence > 68% (non-responders) will be re-randomized to either continued individualized adherence feedback or individualized adherence feedback augmented with problem-solving skills training via telehealth. Thus, there are three intervention strategies embedded in this SMART: #1 control, #2 treatment, and #3 adaptive treatment (intervention appropriate to patients’ response to initial treatment). The primary outcome is electronically monitored adherence. Secondary outcomes include asthma control/severity, lung function, and healthcare utilization. If the aims of the project are achieved, this study would have a large impact on pediatric asthma, with the potential to change clinical practice for treating non-adherence. The SMART design will also allow us to identify patients who are most likely to respond to interventions and assess treatment response to allow us to efficiently deliver the right care to the right patient at the right time in the future.

NIH Research Projects · FY 2025 · 2025-09

The development of next-generation therapeutics (genomic, cellular, and small molecule) in Fragile X Syndrome (FXS) is hindered by three knowledge gaps: 1) poor understanding of the developmental trajectory of brain dysfunction, 2) limited ability to account for phenotypic heterogeneity, and 3) inadequate validation of how translational models represent the human condition. Our team, established over a decade ago, has developed a stakeholder-informed translational research program dedicated to understanding FXS brain physiology across cellular, circuit, and systems levels. Our previous Centers have led the field in establishing an empirical framework for characterizing shared electrophysiological biomarkers in adults with FXS and the Fmr1-/- knockout (KO) mouse. To address the present gaps, our team proposes to synchronize human pediatric and mouse developmental studies, leveraging recent discoveries and our well-established methods to establish translational parallels. The proposed work addresses key limitations our of previous work while directly addressing all three RFA-HD-25-002 specific points of interest: 1) development of translatable biomarkers, 2) resolving phenotypic heterogeneity, and 3) identification of novel mechanisms. The NIH Center mechanism provides an ideal platform for our “Cells to Circuits to Systems to Community” approach, marked by close collaboration with scientists and community stakeholders, and facilitating three linked projects that span patient-centered research to in vivo animal experiments to layer-specific micro-circuit physiology: Project 1 (Erickson/Schmitt, Cincinnati Children’s) will study FXS youth aged 2-17 years to determine age-related changes and developmental trajectory of translatable biomarkers in FXS using resting, evoked auditory, and cognitive paradigms and evaluate heterogeneity by considering the impact of sex, age, FMRP expression, and EEG signatures on cognitive deficits and drug response. Project 2 (Binder/Razak, UC Riverside) will study brain activity using surface multi-electrode array (MEA) and depth array in vivo electrophysiology in the Fmr1 KO mouse across developmental stages using resting and evoked auditory paradigms in parallel with human studies and examine novel mechanistic approaches to address circuit hyperexcitability using genetic models and pharmacologic probes impacting glutamate NMDA receptors GluN2C/D and T-type Ca+ channels. Project 3 (Huber/Gibson, UT Southwestern) will determine synaptic, cellular and microcircuit mechanisms by which GluN2C/D drives excitability of circuits and alterations in thalamic-driven laminar activity and synchrony as well as study the maturation of multi-site, network-level function in Fmr1 KO mice using tetrode-based in vivo recordings during cognitive paradigms used in human studies. Across Projects, phenotypic variability will be examined across a large sample of human participants, mouse background strain, sex, age and FMRP expression. This comprehensive approach aims to accelerate treatment discovery in FXS by providing a foundation informed by development, variability, and mechanistic insights to advance translational medicine goals in FXS.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY The goal of this R35 application is to capitalize on new mouse models of epilepsy that my lab has developed to determine whether, when and under what conditions epileptogenesis can be treated and reversed. Studies are designed to address fundamental questions about the basic mechanisms of epileptogenesis and to advance the development of AAV-mediated gene therapy approaches to bring new treatments to patients with intractable epilepsy caused by mTOR pathway mutations. The present proposal is a culmination of almost two decades of research examining basic mechanisms of epileptogenesis in mouse mTORopathy models. The mTORopathies are a class of developmental diseases caused by mutations in the mTOR pathway. Disease-causing germline and somatic mutations have been identified in more than a dozen mTOR pathway genes, causing focal brain malformations, autism and epilepsy. Tuberous sclerosis complex (TS) and focal cortical dysplasia type II (FCD), the focus of the current proposal, are among the most common conditions. Studies take advantage of key features of the mouse mTORopathy models we have developed. Notably, AAVs are used to experimentally control the epilepsy-causing mutation (deletion of an mTOR pathway gene), allowing us to regulate the number, position and cell type specificity of mutant cells, thus modeling the genetics, focality and cellular mosaicism characteristic of mTORopathies. Moreover, we can combine these models with other technical innovations, such as optogenetics, chemogenetics, calcium imaging and diphtheria toxin-mediated cell ablation. These capabilities put us in position to answer fundamental questions in the epilepsy field. Specifically, we will examine epilepsy reversibility by determining whether and when ablating the mutant cells that cause epilepsy will cure the disease. While focal brain lesions cause epilepsy in mTORopathies, it is not known whether the lesions alone drive the seizures or whether epilepsy can be sustained by recruitment of cells outside the focal lesions. Time course experiments with this ablation approach will reveal if there is a critical period for efficacy, while real-time imaging experiments will examine recruitment of surrounding tissues. Manipulating the cellular distribution of mTOR mutations in the models will reveal whether this feature of the disease is clinically relevant. Mouse experiments will utilize well validated – but non translatable – approaches to set the bar for therapeutic interventions, while new – translatable – AAV vector gene-therapy constructs will be developed and tested in animal models and resected patient tissues. At the completion of the proposed studies, we plan to have developed optimized AAV vectors for treating mTORopathies, and generated data to inform when treatments need to be given, the extent of brain tissue targeted, and how treatments need to be adapted for different patient subclasses.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Eosinophilic esophagitis (EoE) is a chronic allergic inflammatory esophageal disorder, exhibiting clinical symptoms (e.g., vomiting, pain, dysphagia, food impaction) and histologic features of esophageal eosinophilia, epithelial hyperplasia, and dilated intercellular spaces, which are indicative of impaired barrier function. EoE is highly heritable, yet most genetic research has focused on common variants identified by genome-wide association studies (GWAS), particularly implicating genes like calpain 14 (CAPN14) and thymic stromal lymphopoietin (TSLP), both expressed in esophageal epithelial cells. Our research is a significant extension of these studies, utilizing whole-exome sequencing (WES) on 1,464 patients with EoE from unrelated families, and revealed a notable enrichment of rare genetic variants in the nucleotide-binding oligomerization domaincontaining 2 (NOD2) gene. NOD2 , a key regulator of inflammatory responses, is expressed in both eosinophils and esophageal epithelial cells, suggesting its pivotal role in EoE pathogenesis We central hypothesize that gain-of-function NOD2 variants contribute to esophageal epithelial dysfunction and eosinophil activation, thereby exacerbating inflammatory responses in the type 2 immune environment. Our study comprises three specific aims outlining an integrated set of multidisciplinary studies with the necessary statistical and experimental support to evaluate the impact of the effects of NOD2 variants in EoE. Aim 1 investigates how NOD2 variants impair epithelial barrier integrity, focusing on the mechanistic pathways, including NFKB signaling and reactive oxygen species (ROS) production, that drive dysfunction in esophageal epithelial cells. We will use human induced pluripotent stem cell [iPSC]-derived esophageal organoids with CRISPR/Cas9 genome editing to model the effects of NOD2 variants in epithelial cells. Aim 2 explores the impact of NOD2 variants on eosinophil activation and their interactions with epithelial cells. We hypothesize that these variants enhance eosinophil activation and survival, further amplifying inflammation through altered intercellular signaling. By analyzing the gene expression profiles of cocultured eosinophils and epithelial cells, we aim to clarify how NOD2 variants contribute to EoE pathology via their influence on both cell types. Aim 3 examines the combinatory effects of NOD2 variants with other EoE-associated genetic variants, both common and rare. This aim will help establish whether specific combinations of genetic variants significantly increase the risk for EoE, providing insights into genetic susceptibility and pathogenesis. Our multidisciplinary approach combines genetic, molecular, and functional assays, utilizing advanced disease models, including iPSC-derived gastrointestinal organoids with genome editing of iPSCs by CRISPR/Cas9. This integrative design will provide deeper insights into the genetic mechanisms driving EoE. This research addresses a critical unmet medical need, potentially leading to more personalized and effective treatments for EoE and related gastrointestinal conditions.

NIH Research Projects · FY 2025 · 2025-09

Project Summary The overall goal of this project is to target and mechanistically interrogate polyamine metabolism in leukemia stem cells (LSCs) with the long-term objective of developing LSC directed therapy to improve the outcomes of patients with acute myeloid leukemia (AML). AML is a devasting disease with a 30% five-year survival rate, in part due to high relapse rates. While most AML patients achieve a clinical response, therapy resistant AML cells often persist post treatment. A subset of therapy resistant cells called leukemia stem cells (LSCs) exist in residual disease and are responsible for driving disease relapse in most AML patients; making it is critically important to develop new therapies to target the LSC population to improve AML patient outcomes. LSCs have unique and targetable metabolic requirements. Using mass spectrometry-based metabolomics analysis, we have shown that the metabolite levels within LSCs isolated from primary AML patients are distinct from their normal counterpart, hematopoietic stem and progenitor cells (HSPCs). Polyamine levels were significantly increased in LSCs compared HSPCs. Pharmacologic depletion of polyamine levels showed that LSCs are reliant on polyamine metabolism for their survival. Polyamines are cationic compounds containing at least two amino groups, that have pleiotropic effects on cell function and importantly have not been examined in LSC biology. Mechanistic interrogation of polyamine biology in LSCs revealed that polyamines are used to regulate protein synthesis. Polyamines regulate mRNA translation by serving as a precursor for the post-translational modification, hypusination, which is essential for the activity of the eukaryotic initiation factor 5A (eIF5A). eIF5A is a translation elongation factor that is important in the synthesis of a subset of proteins that contain amino acid sequences that are less efficient at forming peptide bonds such as proline rich regions. Thus, polyamines may mediate LSC function through the regulation of protein synthesis of select mRNAs that are essential for LSC survival. Based on our preliminary data, we hypothesize that polyamines are required for LSC function through their role in regulating eIF5A-dependent protein synthesis. We will examine this hypothesis by determining the molecular and biological consequences of polyamine depletion in LSCs. (Aim 1). Further, mechanistically dissect the role of polyamines in regulating eIF5A dependent protein synthesis in LSC function (Aim 2). Through these highly translational studies, we expect to determine the importance of polyamine supported eIF5A hypusination on LSC function and evaluate the therapeutic potential of targeting polyamine metabolism in AML. Our studies will reveal novel LSC biology, the mechanisms by which polyamine depletion/reduced eIF5A hypusination decrease LSC function and define potential pathways the mediate resistance to polyamine depletion in LSCs. We anticipate the proposed experiments will serve as the foundation for a future clinical trial targeting polyamine metabolism in AML.

NIH Research Projects · FY 2025 · 2025-09

Abstract. Newborn babies are highly vulnerable to infection by normally innocuous commensal bacteria. This includes the ubiquitous Gram-negative bacterium Escherichia coli, which is the leading cause of neonatal sepsis. Our preliminary studies show susceptibility in this early life developmental window extends to mice, since neonatal mice uniformly succumb to even low dose infection. Despite shared vulnerability, an interesting distinction is the uniform susceptibility of neonatal mice to E. coli infection – which is in sharp contrast to only 30-50 cases in every 100,000 live birth human babies. We reasoned that if the newborn status alone confers susceptibility to E. coli infection, this distinction begs the question as to why invasive E. coli infection does not occur more frequently in human babies given their universal early life exposure to this ubiquitous commensal. In other words, instead of investigating why babies are susceptible to E. coli, perhaps the more relevant question is why infection does not occur more frequently beyond this small fraction of all E. coli exposed human babies. In this regard, E. coli colonization represents an inherent, but underappreciated, difference between humans and standard laboratory mice. Commensal E. coli can be found in the feces of nearly all individuals, but is completely devoid in mice reared under specific pathogen free laboratory conditions. Our ongoing studies show high titer anti-E. coli “natural” antibodies in healthy adult individuals, their persistence during pregnancy and neonatal cord blood specimens. This “natural” classification indicates anti-E. coli antibodies are physiologically primed by commensal colonization, without infection or immunization. Comparatively mice devoid of commensal E. coli show only background anti-E. coli IgG titers, which rise sharply after experimental colonization using non-pathogenic probiotic strains such as Nissle 1917. In turn, anti-E. coli IgG is efficiently transferred from Nissle colonized mice to their neonatal offspring, which overrides their susceptibility to E. coli infection. Thus, our overall hypothesis (scientific premise) is that vertically transferred maternal immunity overrides the inherent susceptibility of newborn infants to E. coli. By extension, babies that do develop E. coli infection have quantitative or qualitative defects in vertically transferred anti-E. coli immunity. This groundbreaking hypothesis is supported by ongoing studies showing reduced anti-E. coli IgG levels in newborn blood spot specimens of babies that develop E. coli sepsis, and will be further investigated in combination with the aforementioned preclinical platform of E. coli colonization primed immunity that extends to vertically transferred protection in neonatal offspring through the following aims: investigate anti-E. coli antibody titers, along with infant variables including sex and gestational age in vertically transferred IgG function (Aim 1), establish the specific Fc receptor (FcR) and neonatal cell type(s) responsive to IgG that protect against E. coli infection (Aim 2), and evaluate breadth of protection primed by Nissle colonization against E. coli strains that cause neonatal infection (Aim 3).

NIH Research Projects · FY 2025 · 2025-09

Abstract Rotaviruses (RVs) cause severe diarrhea in young children. The current live RV vaccines show reduced efficacy in low- and middle-income countries (LMICs), ranging from 40% to 60%. Consequently, RV infections continue to claim ~130,000 lives annually and cause enormous morbidity, calling for a new generation of RV vaccines with improved effectiveness. Reasons for the diminished vaccine efficacy in LMICs include several factors that disrupt gut’s health and reduce the replication capacity, and thus the efficacy, of live RV vaccines. In response to these challenges, a non-replicating RV vaccine presents itself as a viable option for enhancing effectiveness in LMICs. To address this need, we have invented pseudovirus nanoparticles (PVNPs) named S-VP4e. They consist of a calicivirus inner shell and multiple RV spikes (VP4e) on the surface. The exposed RV spikes formed by VP4e facilitate RV attachment and entry. Antibodies elicited by natural RV infections are largely specific to VP4e and are predominantly neutralizing and protective, supporting VP4e as an ideal vaccine target. The bioengineered S-VP4e PVNP is self-assembled, easily produced, stable, highly immunogenic, and protective against diarrhea caused by RV challenge, making it an excellent RV vaccine candidate. Furthermore, the non- replicating nature of the PVNP will bypass the replication issue of live vaccines and avoid the intussusception risk associated with the replication of live RV vaccines, ensuring better efficacy and safety. Additionally, its low- cost production offers improved cost-effectiveness for LMICs. As a proof of concept, we have demonstrated that a trivalent vaccine comprising three S-VP4e PVNPs, representing the three predominant RV P types (P[8], P[4], and P[6]), elicited high and balanced neutralizing antibody titers against the three major RV types after intramuscular injection. Moreover, it effectively protected mice from diarrhea caused by challenge with RVs. In this application we will evaluate the trivalent PVNP vaccine for oral administration, using U-Omp19 as an adjuvant to enhance mucosal immunity. U-Omp19 is a protease inhibitor that protects vaccine antigens from degradation in the gastrointestinal tract while triggering vaccine specific mucosal immune responses. The safety, immunogenicity, and protective efficacy of the PVNP vaccine will be assessed using both mouse and highly relevant gnotobiotic (Gn) pig models. This non-replicating PVNP vaccine with broad protection represent an excellent candidate for the next generation of RV vaccine. The outcomes of this project will demonstrate the vaccine's utility and provide crucial data for advancing human clinical trials. Two major lines of experiments will be conducted. First, we will assess the PVNP vaccine for its cross-P type mucosal and systemic immune responses, neutralization, and protective efficacy in mice after oral administration, using U-Omp19 as an adjuvant. Second, the mucosal and systemic immune responses and protective efficacy of the vaccine will be evaluated in Gn pig human RV challenge models. With strong preliminary data and an outstanding track record of the research team, we are well-positioned to successfully achieve the goals of this application.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Poor disease self-management is a significant issue in pediatric IBD, with up to 88% of adolescents demonstrating nonadherence to medication. This is a significant concern given that 1) the risk relapse in IBD is 5.5 times greater in nonadherent patients than in adherent patients; 2) the annual costs of health care in nonadherent IBD patients can be up to 12.5% higher; and 3) the estimated annual cost of nonadherence in US health care overall is $300 billion. We have identified numerous factors that interfere with adolescents’ ability to demonstrate optimal self-management of their illness. Although these barriers are amenable to intervention, many adolescents lack the self-management skills to overcome them on their own, and clinic-based self- management efforts have proven unfeasible on a large scale. Through our intervention research in pediatric IBD, we have developed and demonstrated the efficacy of a multicomponent self-management intervention. However, the proportion of the IBD population that could receive this type of treatment is restricted due to limited access (e.g., lack of available trained clinicians, distance between patient homes and treatment facility). In collaboration with key stakeholders (i.e., patients, parents, and providers), we have addressed these accessibility issues by iteratively developing an IBD-specific Self-Management Assistance for Recommended Treatment (SMART) mHealth application, which can be used by all key users to address patient self- management needs. With funding from the Clare Foundation we have adapted evidence-based content developed via our prior R21 (HD074842) for a mobile app developed for both iOS and Android operating systems. The SMART IBD App prompts patients to complete daily diaries assessing disease activity and functional ability, tracks medications and symptoms, personalizes reminders for medication and appointments, provides graphical feedback and IBD-specific education via static content and animated videos, and includes self-management behavioral health challenges using gamification strategies. It is integrated with our electronic medical record, Epic, so providers can access critical self-management data for treatment planning. In this project, we will test, via a Phase IIb randomized controlled clinical trial, the effect of the SMART IBD App on medication adherence, disease activity, functional disability, and health-related quality of life in adolescents with IBD compared to Attention Control. A total of 70 patients and their parents will be enrolled. This study will have a significant impact on public health by providing greater access to evidence-based self-management support to a large proportion of patients who otherwise would not receive this intervention. Given the health and economic impact of poor self-management in IBD, this study is timely and important, as it has the potential to positively impact IBD adherence and health outcomes and serve as a model for self-management intervention across pediatric populations.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Candidate: Patricia Vega-Fernandez, MD MSc RhMSUS is a pediatric rheumatologist whose overarching career goal is to improve the outcomes of children affected by autoimmune rheumatologic diseases through a career as an independent translational scientist focused on biology-based imaging and clinical research to support the development of personalized treatment approaches. Dr. Vega-Fernandez, an Associate Professor of Pediatrics at Cincinnati Children’s Hospital Medical Center (CCHMC), has conducted multiple studies focused on addressing the feasibility, reliability and validity of musculoskeletal ultrasound (MSUS) in Juvenile Idiopathic Arthritis (JIA). The proposed R03 project builds on the skills and knowledge gained by Dr. Vega-Fernandez through the KL2 award. Moreover, this R03 will provide her the research and professional skills needed to conduct fundamental research, secure funding, and advance her career as an independent investigator. Mentors/Environment: Dr. Vega-Fernandez and her mentors, Hermine Brunner MD MSc MBA, Tracy V Ting MD MSc RhMSUS, Mekibib Altaye PhD, and Arthur Meyers MD, have assembled a strong team of advisors from CCHMC. The candidate will leverage several of the Center for Clinical & Translational Science and Training (CCTST) cores at the University of Cincinnati to support the proposed research activities. Research: Chronic musculoskeletal pain (CMSKP) is a common problem in childhood. Children with CMSKP are at increased risk for mood disorders, school absences, social isolation, and poor health-related quality of life. Frequently associated with CMSKP are childhood conditions such as juvenile fibromyalgia (JFM) and JIA. While both conditions frequently present with similar symptoms such as joint pain and swelling, there are important differences in the pathophysiology of these diseases: JIA is an autoimmune condition characterized by joint inflammation, while JFM is a non-inflammatory disorder where neurobiologic disruptions results in chronic pain and transient soft tissue and joint swelling. Diagnosis of arthritis in children is based on the presence of joint swelling and limited range of motion and/or tenderness on palpation. Clinical assessment of active arthritis in children is subjective and has low inter-rater reliability. MSUS can objectively inform the presence of joint inflammation. There is a knowledge gap on the clinical significance of MSUS findings in children with CMSKP. The central hypothesis is that the MSUS of ten joints (MSUS-10) score will serve as a diagnostic imaging tool to differentiate inflammatory from non-inflammatory components of joint pain in pediatric CMSKP with focus on JIA and JFM. This hypothesis will be tested with two specific aims: 1) Determine the accuracy of MSUS-10 in identifying the presence of joint inflammation in children with JFM and JIA in the setting of CMSKP; 2) Establish the clinical impact of the MSUS-10 on immediate diagnosis and treatment strategies in JIA children with CMSKP. Expected outcomes are significant in establishing MSUS as an accurate diagnostic imaging tool that complements medical decision-making processes regarding treatment plans in children experiencing CMSKP.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Following ac+va+on, CD4 T cells differen+ate into one of three main effector linages. The Th1, Th2 and Th17 lineage cells are generated in response to specific class of pathogens and are necessary to combat those infec+ons. While we have comprehensive understanding of the specific cytokine cues as well as master transcrip+on factors that are cri+cal for differen+a+on of each of these dis+nct T cell effector lineages, the innate sensing mechanisms that dictate specific differen+a+on of each of the T effector lineages are not clear. The pathogens that drive Th2 responses are fairly dis+nct but the Th1 and Th17 inducing pathogens share many structural similari+es. Classical paFern recogni+on receptor ac+va+on by microbial ligands leads to genera+on of inflammatory cytokine cues in the form of TNF, IL-6, IL-12, IL-1b, etc but we do not understand how CD4 T cells integrate specific cues to generate a Th1 vs Th17 response. We posit here that host sensing mechanisms that extend beyond the classical paFern recogni+on receptors might be involved in genera+ng unique cytokine cues that specifically dictate genera+on of Th1 vs Th17 lineage cells. Our previous work has demonstrated that following sensing of pathogens, conven+onal DCs can induce priming of naïve pathogen specific CD4 T cells in vitro and the quality of the T cell response is dictated by the nature of the priming pathogen. Specifically, s+mula+on of conven+onal DCs with the gut pathogen C. roden)um drives Th17 differen+a+on as opposed to L. monocytogenes, sugges+ng a DC-dependent mechanism for Th17 differen+a+on. Transcrip+onal profiling of DCs upon exposure to Th17 inducing (C. roden)um) versus non-inducing pathogens (L. monocytogenes and S. aureus) and discovered that C. roden+um induced ac+va+on of the PERK pathway of unfolded protein response (DC- UPR) that was integral to its ability to induce Th17 responses. C. roden+um’s ability to induce DC-UPR was shared by its counterparts, the human enteric pathogens, EHEC and EPEC sugges+ng that this property was evolu+onarily conserved in both the microbes and the hosts. More importantly, several autoimmune diseases caused by Th17 cells are associated with the Unfolded protein response sugges+ng an integral link between ER stress and outcome of T cell responses. In this applica+on we propose to inves+gate the importance of this UPR induc+on from the perspec+ve of the microbe as well as the host and dissect the molecular mechanisms of induc+on of the UPR and its impact on Th17 priming and differen+a+on. In aim 1, we will inves+gate the role of DC-UPR in driving tailored immune responses against Th17 inducing pathogens. In aim, 2 we will iden+fy and characterize the nature of the C. roden+um ligand that ac+vates the PERK pathway of UPR in DCs. In aim 3, we will inves+gate the conserved nature of the UPR pathway in human enteric pathogens and human myeloid cells. Successful comple+on of these aims will add to our fundamental understanding of genera+on of Th17 responses and will provide us with tools to either increase or mi+gate development of Th17 response to enhance protec+on against pathogens or protect against auto-immune inflamma+on.

NIH Research Projects · FY 2025 · 2025-09

Up to 60% of adolescents and young adults (AYAs) with cancer are non-adherent to cancer treatment medications and thus at an increased likelihood of experiencing poor health outcomes like relapse and death. Improving adherence has the potential to improve health outcomes among AYAs with cancer but interventions that address the challenges this population faces to adhering to a medication regimen are lacking. AYAs with cancer endorse a range of challenges to taking their medication (e.g., forgetting, lack of motivation) that vary widely across individuals. We hypothesize that improving adherence will require a personalized intervention capable of teaching AYAs the specific behavior change techniques (BCTs) they need to overcome their challenges to adherence. To test this hypothesis, we developed SUMMIT (SUpporting Medication Management with Individualized Treatment). SUMMIT is the first tailored adherence-promotion intervention designed specifically for AYAs with cancer and directly aligns with the NCI’s special interest in “personalized approaches to improve adherence” (NOT-OD-24-146). Preliminary data from our NCI R21-funded pilot trial support SUMMIT’s feasibility, acceptability, and usability and suggest it has the potential to improve medication adherence. Now, the aim of this R01 is to advance the NCI’s goal of “testing of interventions addressing… treatment adherence” (PAR-24-072) by evaluating the efficacy of SUMMIT as compared to uniform standard of care via a randomized clinical trial. We will recruit AYAs receiving care from pediatric and adult hospitals and will randomize 160 AYAs from these sites to SUMMIT or uniform standard of care. AYAs will use an electronic adherence monitoring device to store their medication and we will use these data to evaluate the hypothesis that SUMMIT results in greater improvements in electronically-monitored medication adherence than uniform standard of care. We will also conduct secondary analyses to evaluate potential group differences in quality of life and outpatient encounters and exploratory analyses to identify obstacles and facilitators to scaling-up and spreading SUMMIT.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Obesity is a common, pervasive, chronic disease often beginning in early childhood. Although infant growth characteristics are key signals of lifelong chronic disease risk, there are no current guidelines about how best to screen for infant growth patterns associated with future obesity risk. Several growth metrics have been used in research settings to evaluate patterns of infant growth using attained weight or BMI z-scores, changes in attained weight-for-age (WFA) z-scores, and modeled trajectories of weight or body mass index (BMI). However, each of these metrics suffers from imprecision or difficulty being implemented in pediatric clinical practice. In particular, the frequently-used metric of change in WFA >+0.67 z-score units, dubbed “rapid infant weight gain”, makes incorrect assumptions about infant weight gain, is inconsistently defined, and is often misinterpreted. In 2009, the WHO published globally-representative infant weight velocity (WtVel) standard distributions of normal physiological growth by sex, age (monthly to 24 months), and the specific time interval between measurements (1, 2, 3, 4 or 6 months). These WtVel standard distributions account for changes in expected growth rates as infants age yet are rarely used in either clinical practice or research, largely because they are impractical to apply (e.g., 194 different reference charts, one for each combination of interval length, sex, and age). Inappropriate or poorly-specified infant growth metrics or misinterpretation of metrics by clinicians threaten our ability to identify infants at risk of developing early-onset obesity. Thus, we have an urgent need to rethink how infant growth is assessed and interpreted both in clinical practice and in research settings. With the advancement in computational capacity in electronic health records (EHR) systems, it is now necessary to examine the clinical performance of more complex research-based infant growth velocity metrics. The present study aims to assess the feasibility and predictive validity of various metrics of infant growth velocity among 1.8 million children from a large national clinical EHR dataset. In Aim 1, we will compare the practical application and demographic inclusivity of 1) WHO WtVel z-scores, 2) modifications to these WHO methods (WtVel-M), 3) infant peak weight velocity (PWV), 4) infant peak BMI, and 5) change in WFA z-scores using individual longitudinal clinical data. In Aim 2, we will test the ability of each metric to predict obesity by age 5. We will also externally validate the predictive performance of each metric using an independent EHR dataset. By the completion of this study, we expect to have rigorously evaluated both existing research-based and novel metrics of infant growth, characterizing their generalizability and validity to predict childhood obesity. Our findings will be a critical step towards evidence-based tailored risk prediction of childhood obesity in the clinical setting, bridging the gap in evidence between research-based growth standards and clinical practice.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Preterm birth is associated with decreased nephron endowment resulting in increased risk of chronic kidney disease and end-stage kidney disease. Human nephrogenesis is completed prior to birth at 32-36 weeks gestation. The majority of nephrons are formed through the poorly understood process, lateral branch nephrogenesis (LBN). Nephron acquisition by LBN is unique to primates and occurs after 22 weeks gestation in the human. During LBN, an elongating, non-branching ureteric bud (UB) tip induces nephron development with each nephron directly connected to the ureteric stalk. In contrast, mice do not undergo LBN. Instead, mice generate nephrons exclusively through branch phase nephrogenesis (BpN) wherein the UB undergoes branching morphogenesis with each branch generating approximately two nephrons. The rhesus macaque is the only known model that undergoes LBN like the human and is thus the sole model in which to define signaling pathways controlling LBN. The central hypothesis is that the cell composition and spatial cell-cell signaling in the LBN niche are distinct from BpN and negatively altered by preterm birth. The long-term goals are to leverage the molecular mechanisms enabling LBN to develop strategies to maintain and/or extend LBN-supporting signaling pathways in preterm infants. The objective of this proposal is to identify the progressive compositional, regulatory, and spatially defined signaling environments in mid to late-gestation primate nephrogenesis. This will enable the development of novel approaches to maintain or extend the LBN process postnatally in preterm infants. The rationale is that the unique rhesus tissues already available and to be obtained through this proposal provide the optimal resources to define critical cellular and signaling changes that occur during human kidney development at single-cell and spatial resolution. The first aim is to determine compositional signaling pathway shifts and regulatory relationships from mid to late-gestation primate kidneys. Single-cell multimodal transcription and ATAC-seq analyses will be performed from replicate rhesus fetal kidneys in conjunction with powerful new integrative regulatory prediction analyses. The second aim will be to elucidate spatially defined differences in WNT signaling in mid to late-gestation primate kidneys. A key benefit of using rhesus tissue for these mechanistic studies is that the tissue will be handled and fixed through a consistent protocol, thus minimizing variation in RNA degradation that may falsely result in differences between gestational periods. Spatial expression of key genes within the WNT pathway, including WNT ligands, receptors, inhibitors, and targets will be determined in mid to late-gestation kidneys by RNAScope Hiplex assay and again validated in high-quality human archival tissues to ensure conservation. Expected outcomes include a comprehensive cellular and transcriptional atlas of primate kidney nephrogenesis determination of cell-cell signaling and spatial interactions that drive primate nephron endowment.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY LMNA-related dilated cardiomyopathy (LMNA-DCM) is one of the most severe forms of adult-onset DCM and results from heterozygous loss-of-function mutations in Lamin A/C (LMNA), a key nuclear lamina component. Currently, no therapeutic interventions exist for LMNA-DCM, primarily due to an incomplete understanding of its molecular pathogenesis. This proposed study aims to directly compare two long-standing prevailing hypotheses regarding LMNA-DCM pathogenesis, the gene regulatory hypothesis and the structural hypothesis. The gene regulatory hypothesis proposes that Lamin A/C reduction causes relaxation of lamina-associated heterochromatin domains (LADs) causing pathogenic gene de-repression, while the structural hypothesis posits that Lamin A/C reduction weakens the nuclear envelope causing pathogenic nuclear envelope (NE) rupture. No studies have compared these mechanisms directly or examined their potential interrelation, a gap we aim to fill to advance mechanistic understanding of this disease. In preliminary studies, we unveiled that Lamin A/C’s requirement in cardiomyocytes arises during the postnatal fetal-to-adult heart maturation. This maturation period involves the repression of fetal gene programs and strengthening of the nuclear envelope to withstand expanding sarcomeres, both processes potentially relying on Lamin A/C’s gene regulatory and structural roles. Thus, our central hypothesis is that Lamin A/C is essential for maintaining LAD-associated gene repression and/or preventing NE rupture during postnatal cardiomyocyte maturation. To test this hypothesis, we will use two innovative mouse models: one with a cardiomyocyte-specific Lmna knockout (LmnaCKO) and another with a pathogenic LMNA-DCM frameshift mutation (LmnaFS), both of which expose Lamin A/C’s requirement during this maturation stage. Aim 1 will determine whether LmnaCKO and LmnaFS mice develop LAD disruption and associated de-repression of LAD-associated genes during postnatal cardiomyocyte maturation. Aim 2 will determine whether cardiomyocytes of LmnaCKO and LmnaFS mice exhibit NE rupture, independently or concurrently with LAD disruption, during this maturation phase. Aim 3 will assess the contributions of the LAD disruption and NE ruptures to the development of DCM. We will use LINC complex disruption to rescue LMNA- DCM in our mouse models, and examine whether LAD disruption or NE ruptures are primarily responsible for the disease. The anticipated impact of this study is to provide decisive evidence supporting one of these prevailing hypotheses, thereby accelerating the development of therapeutic strategies for LMNA-DCM.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract The frontonasal prominence (FNP) is the keystone of the midface, which gives rise to the forehead, bridge and tip of the nose, the philtrum, and the primary palate. The FNP is populated by cranial neural crest cells (CNCCs) that migrate from the neural tube and differentiate to give rise to structural facial features like bone and cartilage. These processes are tightly regulated by signaling from adjacent FGF8-expressing ectodermal centers that include the midbrain-hindbrain boundary (MHB), frontonasal ectodermal zone (FEZ), and olfactory epithelium (OlfE). Failure of formation and fusion of the FNP contribute to several congenital craniofacial abnormalities including clefting and hypotelorism. The cleft primary palate (cpp) avian mutant is a model for failed FNP development as the FNP never fuses and is severely hypoplastic. The cpp was first characterized in the 1960s, but the pathogenic-causing variant was only recently identified as a 1 base pair (bp) deletion in Epithelial Splicing Regulatory Protein 2 (ESRP2) that causes a frameshift. We are the first to genotype the cpp and study the molecular mechanisms that cause its severe phenotype. In preliminary studies, we have performed bulk RNA sequencing at day 4 (HH23), before phenotypic expression, and revealed several differentially expressed genes (DEGs) relating to CNCC migration (Semaphorin 3F (SEMA3F)), development (Bone morphogenetic protein 4 (BMP4)), and differentiation (Ovo Like Zinc Finger 2 (OVOL2)) in the cpp. This has led us to our central hypothesis that the cpp phenotype is due to altered ESRP2-dependent splicing of target genes essential for regulating CNCC migration to, and/or differentiation within the FNP. ESRP2 is a regulator of FGFR2 alternative splicing into epithelial (FGFR2b) and mesenchymal (FGFR2c) isoforms, which dictate FGF ligand binding specificity. In Aim 1, we will use BaseScope, RNAscope, and immunostaining to investigate how variant ESRP2 affects FGF signaling within the cpp and RCAS::FGFR2b to attempt to rescue the cpp phenotype via overexpression of FGFR2b. In Aim 2, we will investigate candidate genes identified through our bulk RNA-seq data to determine if ESRP2 has an expanded role in regulating genes important for craniofacial development. We will use enhanced crosslinking and immunoprecipitation (eCLIP) to determine if ESRP2 directly binds to regulate craniofacial development genes as well as FGF8-soaked bead implantation to test the alternative hypothesis that ESRP2 modulates the expression of candidate DEGs indirectly via FGF signaling. This proposal will increase the understanding of alternative splicing in craniofacial development, which will further contribute to advancement of diagnoses and therapeutics for midfacial abnormalities.

NIH Research Projects · FY 2025 · 2025-08

SUMMARY Epidemiological evidence establishes obesity as an independent risk factor for increased severity of viral respiratory pneumonias including influenza virus infection. Dysregulated systemic and tissue inflammation is critical to the pathogenesis of both influenza- and obesity-comorbidities. However, the critical processes that govern increased influenza severity in obesity remain undefined. The expansion of white adipose tissue (WAT) along with activation of WAT-residing immune cells inflammation promotes tissue/organ damage severity in obesity. Of note, WAT depots differ in their anatomical location and function—characteristics that are directly linked with the pathogenesis of disease-impacted organs. Using a model that combines obesity and influenza A virus (IAV) infection we reported that obese mice exhibit in greater mortality and worsened lung inflammation and tissue pathology compared to lean controls. Focusing on immune cells, we showed that IAV infection changes epididymal WAT (eWAT; a depot distal to the lungs) and lung immune cell composition and function, and favors accrual of a macrophage (MØ) subset in the lungs that shares a transcriptomic signature with eWAT residing inflammatory MØs. However, although transfer of eWAT MØ from obese into lean IAV infected mice amplified host immune cell infiltration to the lungs, it was not sufficient to increase influenza severity. Thus, eWAT MØ, although important, either do not possess the full extent of pathogenic traits or other WAT-residing immune cells are dominant contributors to influenza severity in obesity. Importantly, the presence of thoracic WAT (tWAT; a depot that is proximal to the lungs) was reported in individuals living with obesity and in obese mice. Notably, adipocytes and MØ within tWAT support IAV replication and produce proinflammatory factors during IAV infection. However, whether the character and function of tWAT and eWAT immune cells, including MØ, differs during IAV infection has not been studied. Our preliminary studies comparing tWAT and eWAT immune cells show that: (i) transfer of tWAT immune cells from obese mice into IAV infected lean mice induces mortality; (ii) tWAT is highly enriched in phenotypically distinct immune cell types; and (iii) tWAT MØ exhibit greater ability to produce proinflammatory cytokines relevant in influenza pathogenesis. Together, our novel data and existing literature support the overarching hypothesis that tWAT MØ become progressively more proinflammatory during obesity and IAV infection, and that activation of tWAT MØ unique pathogenic traits is sufficient to increases influenza severity. To test this hypothesis, we will: (1) Determine cellular traits of tWAT immune cells in obesity and influenza severity; and (2) Determine pathological processes whereby tWAT macrophages exacerbate influenza severity. Given the global increase in the incidence of obesity and viral pneumonias (e.g., Influenza, SARS-CoV-2) our high-risk/high-reward proposal will provide keen insights into previously unexplored processes that govern inflammation-associated disease severity in obesity.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT: Eosinophilic esophagitis is a chronic food antigen-driven allergic inflammatory disease associated with symptoms such as refractory pain. While esophageal eosinophilia is a hallmark of the disease, there is often a disconnect between eosinophil levels and quality of life, as patients in histological remission often have persistent symptoms. Thus, the mechanism of symptom development in EoE is likely more complex than eosinophilic inflammation alone. Recent studies demonstrate that the function of afferent nerves extends beyond their role of relaying nociceptive information back to the CNS. Sensory neurons can directly sense IL-4 and IL-13, two influential cytokines in EoE pathogenesis. These have been shown to activate nerves to produce factors that then modulate immune cell function. One such factor is insulin-like growth factor 1 (IGF-1), which can regulate neuronal sensitization and pain. In allergic inflammation and asthma, IGF-1 has been shown to promote mast cell survival, regulate eosinophil levels, and cause proliferation of epithelial cells. Taken together, evidence is emerging that the peripheral nervous system may contribute to allergic disease pathophysiology as an intricate part of the innate immune response. But the role of the nervous system in EoE pathogenesis has not received much attention. Preliminary data demonstrated that mice with experimental EoE display elevated eosinophil levels, increased non-evoked pain, heightened sensory neuron innervation density in the esophagus, and altered sensitivity of sensory neurons to noxious stimuli. Sensory neuron-specific deletion of IL4Ra (IL4Ra-/-NaV1.8) reduced inflammatory gene expression, histological severity, eosinophil levels, and innervation density in allergic animals compared to their wild-type counterparts. Additionally, the cell bodies of esophageal afferents in the dorsal root ganglia (DRGs) in mice with experimental EoE express elevated IGF-1, but is significantly reduced in allergic IL4Ra-/-NaV1.8 mice DRGs and esophagi. Mechanistically, IGF-1 levels have been linked to IL4Ra activation in multiple cell types. Therefore, we will test the central hypothesis that the activation of esophageal sensory neurons via IL4Ra during allergic inflammation drives increased nerve hypersensitivity and subsequent inflammation via IGF-1 signaling. Transgenic, DREADD, and AAV technology will be used to determine the role of sensory neurons on nociception and inflammation in experimental EoE. Pain-related behavioral assays, in vivo calcium imaging of DRGs, and molecular analyses (qPCR, flow cytometry, IHC, IF) of the esophagus will be conducted. Aim 1 will elucidate the role of peripheral nociceptive neurons and their expression of IL4Ra in a murine model of EoE through chemogenetic manipulation of esophageal neuron firing. Aim 2 will determine whether sensory-neuron derived IGF-1 is necessary to increase esophageal hypersensitivity and inflammation in experimental EoE. This research will be the first to assess the presence of a potential neuroimmune mechanism by which nerves contribute to disease development and symptom severity via expression of IL4Ra and IGF-1 signaling, in EoE.

NIH Research Projects · FY 2025 · 2025-08

PROJECT ABSTRACT / SUMMARY Although CFTR modulator therapies have improved respiratory outcomes for the Cystic Fibrosis (CF) population, the benefit to individual patients can vary widely. In “real world” studies of modulator therapies, patients continue to experience lung function decline, and as many as 1 in 5 receive no therapeutic benefit in FEV1. The factors that determine who has a strong or poor therapeutic response are not well defined, and limit both the current benefit from modulator drugs and future benefit from gene therapies. One proposed but poorly understood factor is the solute carrier SLC26A9, and specifically the risk locus rs7512462 within that gene. This locus is associated with extrapulmonary disease severity in CF, and appears to be linked to CFTR modulator response, though the mechanism of this association and its future impact on CF gene therapy remains unclear. Our long-term goal is to create personalized medicine strategies to optimize care while reducing disparities in disease burden for individuals with airway disorders. The overarching goal of this project is to identify the cellular mechanism and real-world impact of SNP rs7512462 in SLC26A9 on modulator response, and to understand the impact of this interaction on future CFTR-focused therapies. Our central hypothesis for this work is that rs7512462 influences SLC26A9 expression, which is a major determinant of lung function response to modulators via modification of airway ion/fluid homeostasis. We will address this hypothesis through two aims. First, we will recruit individuals with CF and above- or below- average response to modulators based on rate of lung function decline. We will evaluate the relative impact of SLC26A9 genotype on lung function decline in this cohort, and the physiologic impact on the respiratory epithelium using patient-specific ex vivo airway culture. Second, we will determine the mechanism of this interaction through in vitro studies utilizing isogenic cell lines co-expressing risk and protective alleles at rs7512462 with key CFTR variants. We will determine the impact of SLC26A9 genotype on CFTR rescue by modulators and by CFTR gene replacement therapy using an AAV-CFTR construct. We will also assess SLC26A9 as a potential therapy, delivering this gene construct in a similar AAV vector and quantifying its impact on airway epithelial physiology. Through these studies, we will identify the magnitude, mechanism, and real-world relevance of SLC26A9’s influence on patient-specific disease variance, as well as generate proof-of-concept of SLC26A9 as a therapeutic agent. This approach will lead to novel targets to stratify risk and optimize care at the individual level, facilitating precision care in muco-obstructive lung diseases impacted by SLC26A9, including CF.

NIH Research Projects · FY 2025 · 2025-08

Project Summary DeGAUSS (Decentralized Geomarker Assessment for Multi-Site Studies) is an open-source research tool that transforms geocoding and exposure assessment models into FAIR (Findable, Accessible, Interoperable, Reusable) computable exposures, enabling privacy-preserving assessments without sharing identifiable information. The project’s overall objective is to enhance DeGAUSS’s scalability, interoperability, and reusability, making geocoding and geomarker assessment more accessible to all researchers. We will leverage advances in computing for enhanced accessibility and portability by refactoring DeGAUSS using Rust-based geospatial libraries while maintaining an R package as the user interface. This will reduce dependencies on external system libraries, allow for containerless deployment, and simplify reproducible environments. We plan to update geocoding for exact address matching with Safe Harbor compliance. We will develop APIs and interoperable services compliant with modern standards for DeGAUSS. Creating APIs and services that adhere to Swagger standards will enable seamless integration with existing data pipelines. Building an OCI-compliant container with a web-based interface and leveraging WebAssembly (Wasm) to embed Rust and R will provide a secure, portable solution adaptable to various environments. We aim to enhance reusability through standardized data formats and expanded testing. Adopting standardized input/output data formats and expanding testing with diverse datasets will enhance the accuracy, reliability, and applicability of geocoding and geomarker assessment tasks. We plan to increase community engagement and uptake through improved dissemination channels. By developing educational materials, conducting workshops, and collaborating with standards organizations, we will raise awareness, improve usability, and ensure DeGAUSS aligns with evolving best practices, fostering widespread adoption. By achieving these aims, the project will extend DeGAUSS’s impact and sustainability, contributing to a better understanding of environmental influences on health and supporting the NIH’s mission to leverage advanced data tools for improving public health outcomes.

NIH Research Projects · FY 2025 · 2025-08

The aim of our proposal is to deliver intact, functional CFTR using AAV into CF patient cells to relieve the symptoms of CF regardless of the CFTR mutations that they carry. Previous approaches for delivering CFTR using AAV have been conducted using truncated CFTR constructs due to the cargo size limitation of AAV. This approach was not successful, potentially due to dysfunction of the truncated protein. To circumvent this limitation of AAV and deliver a full-length intact CFTR into cells, we have employed intein technology. The CFTR gene is split into two constructs and attached to coding segments for intein, and delivered using two AAVs (a dual AAV approach). This delivered gene is then translated in the cell, and the resulting protein halves are spliced together by inteins to create a functional, full-length CFTR protein. We have tested this dual AAV system using primary airway epithelial cells from six CF patients who do not respond to CFTR modulators and have seen significantly improved CFTR function in vitro. In this proposal, we will optimize delivery of this split CFTR, dual AAV system in airway epithelial cultures from people with CF that do not respond to modulators. This system will also be applied to a CF-relevant mouse model to study efficiency and durability of gene transduction. Since AAV is clinically approved for gene therapy to deliver a therapeutic gene (e.g., Hemophilia gene therapy) and a dual AAV approach was also successfully applied in a recent clinical trial for deaf patients lacking functional otoferlin (OTOF), we expect in the future that our dual AAV with intein may be used to restore CFTR activity in patients with CF, which is especially important for those CF patients who do not respond to CFTR modulators.

NIH Research Projects · FY 2025 · 2025-08

Project summary: Peripherally inserted central catheters (PICCs) have become an integral part of the complex intensive care of critically ill patients in pediatric intensive care units (ICUs). Unfortunately, PICC malposition occurs in approximately 3.6%-5.6% of pediatric patients. It is critically important to frequently assess the position of PICCs, because malpositioned PICCs can lead to a variety of clinical complications, such as thrombosis, cardiac arrhythmia, and inadequate therapy. Chest X-rays (CXRs) are widely used radiographic images for the periodic assessment of PICCs in children. However, assessing PICCs is a labor-intensive, time-intensive task for radiologists. Prior studies have demonstrated the promise of deep learning for automated PICC assessment in adult patients. To date, there are no available methods for pediatric PICC assessment. An accurate, robust, and clinically available tool that can automatically assess PICCs in pediatric CXRs is a critical unmet need. The objective of this proposal is to develop an automated PICC assessment pipeline for pediatric CXRs using deep learning algorithms. To accomplish our objective, we will leverage ~4,000 retrospectively collected CXRs from pediatric ICUs. In the current clinical assessment of PICCs using CXRs, radiologists would typically (1) identify each PICC line, (2) locate the ideal PICC landing zone, and (3) determine whether the distal tip of the PICC is appropriately positioned within the landing zone. By closely mimicking radiologists, we design a novel, transparent, and interpretable three-component PICC assessment pipeline. Specifically, we propose to (Aim 1) develop a topology-preserving deep learning method to segment PICCs, (Aim 2) develop a landmark detection method to locate the ideal PICC landing zone, and (Aim 3) assemble the pipeline and evaluate its performance in detecting PICC malposition using CXRs in pediatric patients. This project is highly impactful to clinical practice. Assessment of PICCs commonly occurs within a procedure called Lines, Drains, and Airways (LDAs) reconciliation, in which radiologists review LDA intensive care devices on CXRs and communicate with intensive care physicians in ICUs to ensure safe patient care. Our proposed pipeline will promote LDA reconciliation between radiologists and intensive care physicians. By taking timely remedies, physicians in ICUs can quickly reposition those malpositioned PICCs back to the ideal landing zone, thereby mitigating potential risks of major clinical complications. Once validated, our pipeline can also be extended to other LDA devices, such as endotracheal tubes or chest tubes. The resulting models will be made available to the community for both research and clinical purposes.