Cincinnati Childrens Hosp Med Ctr

NIH Research Projects · FY 2025 · 2019-08

PROJECT SUMMARY Diseases of the lung are a significant global health burden. To develop the next generation of therapies for diverse lung diseases, system models informed by an ever-increasing repertoire of molecular omics, cellular, spatial imaging, and pathological datasets are desperately needed. The LungMAP DCC is responsible for data collation, re-analysis, and integration; secondary annotation tracking; developing tools to facilitate collection, sharing and data dissemination; operating a web resource for data, expertise, and collaboration; and coordinating activities across the Research Centers (RCs) and Human Tissue Core. The DCC also must facilitate literacy for investigator use of developed tools and best practices for analysis, data provenance and metadata annotation, and engage the larger research community. To cultivate future discoveries by the lung community, we have begun aggregating a diverse collection of single-cell atlases for multiple species and diseases into highly curated lung cell atlases as queryable datasets, with an emphasis on dynamic visualization, figure generation, re-analysis, cell-type curation, and automated annotation. In LungMAP Phase 3, to support the discovery of novel cellular and gene regulatory mechanisms, the LungMAP DCC will assemble, annotate, analyze and distribute multi-omic catalogs of diverse lung diseases, expand and enrich our portal ecosystem and lead community curation efforts. These resources will be deployed for broad reuse throughout the lung research community. Our specific aims are to: 1) Advance lung research, education, cooperation, and scientific dissemination efforts across the lung research community. The LungMAP Administrative Coordination Center (ACC) will coordinate scientific and operational administrative activities to maximize synergies and research opportunities within the consortium and with community partners. 2) Create a multi-omic catalog of lung diseases leveraging LungMAP contributed and community data sources. The DCC will collect, organize and analyze a large collection of LungMAP and community datasets to produce harmonized disease cell atlases and disease focused portals. 3) Develop LungMAP.net into an extendable knowledge base of lung disease network biology. The LungMAP.net ecosystem will expand to serve as a hub for research as a living resource and knowledge base for normal and disease lung modeling through integration of agile AI-driven tools. Thus, the LungMAP DCC will deliver a comprehensive corpus of standardized and integrated lung data and knowledge, to nominate new targets for therapy, new regulators and disease subtypes. 1

NIH Research Projects · FY 2025 · 2019-08

Project Summary/Abstract Rare diseases (RD) and disorders collectively affect up to 30 million Americans. RD research faces key challenges: 1) insufficient knowledge about the etiology, pathophysiology, natural history and epidemiology of the diseases; 2) inadequate or non- uniform case definition and disease classification systems that make diagnosis and epidemiologic assessment difficult; 3) insufficient understanding of the determinants of multiple phenotypes and the relationships between genetic variance and phenotypic manifestations; 4) rarity and geographic dispersion of cases that hampers both access to qualified care and participation in research; 5) a dearth of clinically proven, safe and effective treatments; and 6) inadequate private investment into RD research and treatment. The Rare Diseases Act of 2002 authorized the Office of Rare Disease Research to recommend a research agenda and promote coordination and cooperation among research programs. The Rare Diseases Clinical Research Consortia (RDCRCs) that comprise the Rare Diseases Clinical Research Network (RDCRN) advance the diagnosis, management, and treatment of RDs to enhance clinical trial readiness. The RDCRN Data Management and Coordinating Center (DMCC) must provide state-of-the-art informatics, statistical and epidemiological expertise in clinical research study design and data management in order to support the production of evidence that can support the progression of rare disease clinical research (RDCR). To achieve clinical trial readiness (CTR) throughout the RDCRN, we will continue to lead the DMCC based at Cincinnati Children’s (CCHMC) pursuing three Aims: 1) To advance the methods and practice of RDCR and promote CTR through the effort of our Clinical Research Core (CRC) and Data Management Core (DMC); 2) To maintain and enhance a leading-edge, cloud-based data ecosystem to facilitate the research performed by the RDCRCs and promote data sharing; and 3) To expand research collaboration within the RDCRN and with other stakeholders and disseminate the RDCRN research findings, promoting the RDCRN globally. The DMCC Administrative Core will maintain an efficient organization of the DMCC, support RDCRN governance and promote collaboration across the RDCRN; the Data Management Core will support the RDCRN Operational Environment will facilitate FAIR data sharing in the RDCRN Data Repository; the Clinical Research Core that will advance CTR and best practices in RDCR, and will offer training through a methodology hub; the Engagement and Dissemination Core will implement a broad RDCRN outreach and dissemination program, engage the PAGs in RDCR, and offer opportunities for career enhancement. We rely on a combination of world-class expertise, outstanding infrastructure, state-of-the-art technology and enthusiastic institutional support. We will hasten scientific discovery within the RDCRN and bring new treatments to trial, ultimately promoting health and wellness for RD patients and their families.

NIH Research Projects · FY 2026 · 2019-06

ABSTRACT: Approximately 15-20% of children experience persistent or chronic pain. However, compared to adults, we know relatively little about the mechanisms of pediatric pain development. A basic understanding of nociceptive processing in the immature nervous system is therefore crucial in order to develop more appropriate treatments for pain in children. The developing peripheral nervous and immune systems are functionally distinct from adults. These systems are vulnerable to effects of early life injury which can influence outcomes related to nociception following subsequent injury later in life (i.e. “neonatal nociceptive priming”). We have found that macrophages are a key player in both early life nociception and neonatal nociceptive priming responses after incision injuries. Macrophages were found to retain an epigeneetically driven memory of early life injury that leads to a more pro-inflammatory state such that re-injury causes a prolonged behavioral and physiological responses in the peripheral nervous system. Observed changes in the PNS that underlie neonatal nociceptive priming are blocked by genetic targeting of the nerve growth factor (NGF) receptor, p75, in macrophages, one of the key factors found to be part of the epigenetic memory. New pilot data suggests that mast cells (MCs) may be the source of NGF that sustains the pro-inflammatory state created by the epigenetic memory in macrophages. In addition, sensory neurons also appear to generate an epigenetically driven memory that contributes to the pro-inflammatory environment in the muscles after injury leading to prolonged hypersensitivity. The main goal of this proposal is to determine how the epigenetic modifications in this novel neuro-immune circuit regulates neonatal nociceptive priming. Specific Aim 1 will use our novel ex vivo somatosensory recording preparations and pain-like behavioral assays along with cell specific transgenic approaches to determine the additional epigenetically modified factors in macrophages (e.g. parvalbumin) that regulate neonatal nociceptive priming. Specific Aim 2 will test whether knockdown of neuronally produced cytokines (e.g. interleukin 34) modulates neonatal nociceptive priming using similar approaches with nerve targeted gene knockdown strategies. Finally, Specific Aim 3 will use behavioral analyses and/or ex vivo recording to determine the influence of MCs and MC produced NGF in neonatally incised mice on the prolonged effects to subsequent adolescent incision. These aims will be complemented by calcium imaging analysis of human iPSC derived sensory neurons treated with media from macrophages or MCs to enhance translational potetinal of these studies. These experiments will allow a better understanding of the unique mechanisms by which neuroimmune signaling contributes to neonatal nociceptive priming. These studies will facilitate understanding of the transition from acute to chronic pediatric post-surgical pain, and will allow us to determine the utility of targeting immune cell or sensory neuron memories as a pain therapy for children.

NIH Research Projects · FY 2025 · 2019-05

PROJECT SUMMARY ABSTRACT A more universal vaccine against influenza virus infection is urgently needed. However, a major obstacle limiting more effective and durable vaccines against influenza infection stem from the rapidly shifting nature of viral immune dominant epitopes. Further confounding this obstacle are functional differences in the immunological responsiveness to vaccination and susceptibility of individuals to natural infection primed by prior exposure to influenza antigens. This type of immunological imprinting likely explains the wide discordance in effectiveness of current seasonal influenza vaccines. Considering the near ubiquitous exposure of individuals to influenza virus, together with the wide variability in clinical symptoms from asymptomatic to severe infection and increasingly widespread use of seasonal immunization, immunological imprinting to influenza virus is likely initiated during infancy with the first exposure to natural infection or immunization. Importantly, critical knowledge gaps remain regarding how individuals respond to primary influenza exposure in early life, in the context of natural infection or vaccination, and how a lack of pre-existing immunity effects within-host influenza viral diversity. It is also unclear how this first exposure to influenza impacts the subsequent immunological responsiveness to antigenically identical, similar or discordant influenza epitopes. For infants, the impact of vertically transferred maternal immunity, or that acquired postnatally through breastfeeding, on the quality of ensuing influenza specific immune responses remain unclear. To fill these knowledge gaps, ongoing recruitment of a maternal-infant cohort at Cincinnati Children’s Hospital will be expanded, along with parallel efforts in Mexico City. Both sites have ongoing surveillance providing additional cases of symptomatic primary infection. With the proposed enrollment of more than 2000 pregnant women, our two-site cohort is ideally suited for the proposed studies given our established infrastructure of weekly nasal swabs and symptom reporting, scheduled blood collection to detect asymptomatic and symptomatic influenza, maternal and cord blood and milk sample analyses and detailed evaluations of susceptibility and immunological responses to influenza infection and vaccination in mothers and infants. Our overall hypothesis is that primary influenza exposure in early life impacts the magnitude, durability and breadth of immunological memory to an evolving range of influenza virus antigens and this initial imprint will have a profound effect on subsequent influenza exposures. A team of investigators with complementary expertise in pediatric infectious diseases, epidemiology, maternal-infant cohorts, human B and T cell immunology and influenza virology have been assembled to address our hypothesis by investigating the immunological response of infants to primary influenza virus natural infection compared with immunization (Aim 1), compare the immunological response against an initial exposure to influenza virus via natural infection or immunization in infants (Aim 2), and investigating the impact of pre-existing influenza immunity on virus genetic diversity within individuals (Aim 3).

NIH Research Projects · FY 2025 · 2018-08

A key component of this P30 application is the ‘Personalized Cystic Fibrosis Model System Core’. The models will include the patient-specific cultures derived from intestinal epithelia (enteroids) and the induced pluripotent stem cells (iPSCs) from myofibroblasts. These patient-specific cultures will be used to study CFTR function and fluid homeostasis, and the information obtained will be used towards personalized treatment options. The Personalized Cystic Fibrosis Model System core will develop and bank these models, develop and validate the assays for CF/CFTR research, and distribute the resources to researchers worldwide. The long-term goal of this core is to use models derived from the patient to facilitate his/her personalized therapy. Specific Aim 1. To provide well-characterized CF patient-specific somatic epithelial cultures and iPSC to local, national, and international investigators interested in personalized model systems for CF/CFTR-related research. Specific Aim 2. Establish and provide tissues from CF patient-specific iPSC-derived intestinal and pancreatic tissue. Specific Aim 3. To provide assay services and training to local, national, and international investigators interested in personalized model systems to study the functional involvement of CFTR fluid secretion in intestinal enteroids and spheroids. The Core will create resources and sound services that have important long-term clinical implications toward patient-specific therapies for cystic fibrosis. The core will provide training in a variety of state-of-the-art and innovative techniques, including the development of intestinal stem cell cultures, iPSCs technology, high content microscopy, to name a few, to researchers who will be able to use these techniques to study not only CF/CFTR biology but also other membrane proteins and their associated diseases.

NIH Research Projects · FY 2025 · 2018-07

PROJECT SUMMARY There is a fundamental gap in the availability of cognitive outcome measures that are reliable and sensitive to detecting change among children with Down syndrome (DS). Lack of such outcome measures represents an important problem to interpreting clinical trials aimed at improving the lives of individuals with DS. Without evidence-based cognitive outcome measures, future treatment trials in this population will remain suboptimal due to poor study measures. Despite some promising findings in currently recommended assessment batteries, the evidence base for cognitive outcome measures in DS is limited. Multi-site studies evaluating the reliability and validity of outcome measures are the needed next step towards supporting the evaluation of new pharmaceutical and clinical interventions for children with DS. Working groups convened by NICHD of leading experts in DS led to recommendations of promising measures for use in this population. The prior R01 HD093754 began evaluating the psychometric properties of recommended promising measures and expanded upon those recommendations by evaluating individual variability and accounting for the method of assessment (verbal, nonverbal, computer, parent-report) to support our understanding of performance within a cognitive domain. The proposed renewal continues evaluating the psychometric properties of measures that fill the gaps in cognitive domains (Study 1), extends the natural history of change in performance on measures with annual follow-up visits (Study 2), and refines the measurement of individual variability to include co-occurring medical and mental health conditions across both Studies. The overall objective of this application is to establish the psychometric properties of individual clinical outcome measures in children with DS across the cognitive domains of episodic memory, executive functioning (set-shifting, inhibitory control, working memory), learning and memory, and processing speed. Our rationale for working with this population is that DS is associated with a distinct pattern of cognitive strengths and weaknesses related to their neuroanatomy. Thus, the selection of outcome measures to be evaluated needs to take into account the DS behavioral phenotype. We propose three specific aims: 1) To examine the psychometric properties of selected cognitive outcome measures with children with DS. 2) To evaluate differences in the psychometric properties of the measures as a function of variations in demographics and co-occurring medical and mental health conditions. 3) To characterize the developmental trajectories of cognition and executive functioning. To achieve these aims, 200 children in Study 1 ages 6-17 years with DS will participate in repeated neuropsychological assessments with follow-up evaluations at 2 weeks, 6 months, and 1 year. We will continue to follow at minimum 120 children from R01 HD093754 annually in Study 2. We anticipate that this measurement study will provide critical guidance for future efficacy and effectiveness trials. Our goals are in line with the programmatic objective of the INCLUDE project to assemble a large cohort of individuals, perform deep phenotyping, and study co-existing conditions.

NIH Research Projects · FY 2025 · 2018-07

Crohn’s Disease (CD) is a chronic and debilitating disorder with peak incidence in the second and third decades of life. While considerable progress has been made in optimizing medications to achieve remission, relapse is common and unpredictable. Altered microbiota likely drive gut inflammation and clinical relapses. Microbiota-accessible dietary carbohydrates with beneficial health effects, known as “prebiotics,” hold promise for restoring healthy gut microbiota in CD and preventing clinical relapse. Here, we propose completion of the first studies of the prebiotic human milk oligosaccharide, 2’-fucosyllactose (2’-FL), for maintaining remission in CD. Our overarching hypothesis is that 2’-FL supplementation in CD will be safe and well tolerated, while increasing fecal Bifidobacterium abundance and butyrate in a dose dependent manner. We will test this hypothesis by completing a randomized, placebo-controlled dose-ranging study which began enrollment during the tenure of the current award and includes the following Aims: Aim 1. Define dose dependent safety and tolerability of 2’-FL as a dietary supplement in CD. We will test 1g or 5g 2’-FL compared to 2 gm dextrose placebo as a daily dietary supplement in pediatric and young adult CD patients in stable remission receiving infliximab or adalimumab anti-TNF therapy. Safety and tolerability will be assessed using validated clinical disease activity and gastrointestinal symptom rating indices, and fecal calprotectin. Aim 2. Define dose dependent efficacy of 2’-FL as a dietary supplement in CD. We will utilize our established fecal metagenomic and metabolite profiling assays to test the effect of a range of 2’FL doses upon the gut microbial community and associated metabolic functions with a focus upon butyrate production. Efficacy will be assessed by determining the dose dependent effect of 2’-FL upon increased fecal Bifidobacterium and butyrate abundance. We will account for FUT2 secretor status and dietary fiber intake in the analysis. These studies will have a high impact in the field by providing critical phase I/IIa safety and efficacy data in support of a phase III RCT using our NIH-supported CD clinical research network to test the efficacy of 2’-FL in directly modulating beneficial microbiota and thereby enhancing sustained clinical remission and mucosal healing. Ultimately the proposed studies will promote a fundamental shift in clinical practice towards personalized microbial therapeutic interventions.

NIH Research Projects · FY 2026 · 2018-07

Abstract The cardiac fibroblast and its ability to convert into a myofibroblast for extracellular matrix (ECM) production, ventricular remodeling and the fibrotic response has been an area of recent investigation with important medical relevance. Here, a dual-PI renewal resubmission application is proposed by a developmental cardiac biologist and adult disease-based cardiac biologist to address how the postnatal heart matures in adulthood and then transitions back to a fetal-like program with disease stimulation through direct paracrine crosstalk and ECM feedback. During the past funding cycle of this award, we identified key regulatory relationships that exist between myocytes and fibroblasts in both the developing and diseased adult heart, whereby the ECM and transforming growth factor-β (TGFβ) served as an integrating platform between these 2 cell-types. We have also observed that fibroblast-expressed GDF10 and the epidermal growth factor (EGF) family member pleiotrophin (Ptn) mediate critical crosstalk between fibroblasts and myocytes. Here, we will investigate the hypothesis that TGFβ is a myocyte selective maturation factor that controls fibroblast activity in generating an effective ECM within the postnatal heart that also underlies adult disease, and that parallel Ptn and GDF10 signaling crosstalk further regulates fibroblast proliferation and promotes cardiomyocyte hypertrophy in development and disease. The dual-PI renewal application has 3 specific aims: 1) To examine how cardiomyocyte generated TGFβ1/2/3 and its subsequent signaling to cardiac fibroblasts underlie ECM neonatal maturation and adult ventricular remodeling, 2) To examine how cardiac fibroblast generated ECM regulates developmental cardiomyocyte maturation and adult heart remodeling, in part through TGFβ1/2/3 sequestration/release, and 3) To examine the function of the cardiac fibroblast-secreted growth factors Ptn and GDF10 in postnatal heart development and adult injury. Collectively, these specific aims will uncover novel signaling mechanisms that underlie heart maturation just after birth and determine how these mechanisms are redeployed in disease. Thus, the impact of this program will be the identification of novel signaling mechanisms and effectors that can be therapeutically targeted in human heart disease to positively effect cardiac remodeling and longstanding fibrosis, with added implications for treating congenital malformations and developmental growth abnormalities.

NIH Research Projects · FY 2026 · 2018-06

PROJECT SUMMARY Enteric bacterial infections remain one of the greatest public health challenges worldwide and deciphering the mechanisms that protect against infection will enable development of new treatments. Intestinal tissues are in constant direct contact with diverse beneficial and pathogenic microbes, highlighting the need for orchestrating complex microbial signals to sustain protection against infection. Intestinal epithelial cells (IECs) reside at the direct interface between intestinal pathogens, beneficial commensal bacteria, and intestinal immune components. However, despite continuous exposure to diverse microbes, the mechanisms regulating how IECs integrate microbial-derived signals to mount protective host responses to pathogens are not well understood. The goals of this proposal are to interrogate how specific commensal bacterial-derived metabolites are sensed by IECs to protect against pathogenic infection. Employing Citrobacter rodentium, a murine model of human enteropathogenic Escherichia coli infection, our studies have identified that microbiota-derived products protect against intestinal damage and enteric bacterial infection. Our epigenetic analyses for this project led to identification of new commensal bacterial-derived metabolites that can directly regulate IECs and prime host defense against pathogenic bacterial infection. Employing an exciting array of transgenic animals, pathogenic and commensal bacterial strains, and human intestinal organoids, three specific aims are proposed that will (i) decipher how the host calibrates intestinal barrier function by sensing newly-identified commensal bacterial- derived metabolites, (ii) investigate metabolite-dependent regulation of enteric infection, and (iii) interrogate how metabolism of dietary components by commensal bacteria prime the epigenome and enhance host response to pathogenic bacteria. Defining pathways that integrate commensal and pathogenic signals will provide a framework to test the therapeutic potential of manipulating commensal bacterial-derived metabolites to promote antibacterial immunity.

NIH Research Projects · FY 2025 · 2018-04

Abstract Congenital muscular dystrophies (CMDs) are a group of genetic disorders that lead to neuromuscular degeneration and profound muscle wasting with early morbidity and mortality, often in adolescence. The more common mutations leading to CMD are in the LAMA2 gene, although mutations in the 3 genes encoding collagen VI (COL6A1, COL6A2 and COL6A3) is next most common where it leads to Ullrich and Bethlam myopathy. While these CMDs have been studied at the molecular levels for over 3 decades now, the field still lacks an appropriate therapy to treat afflicted individuals. The field also lacks a complete understanding of how this disease is initiated and propagated, although the overarching mechanism appears to be associated with how the skeletal muscle myofiber attaches to the overriding basement membrane. Here we will attempt to elucidate the greater underlying molecular and cellular mechanisms of Ullrich CMD by using a mouse model of Col6a2 deficiency. We hypothesize that detachment of the myofiber from its basement membrane due to collagen VI deficiency is sensed by integrin complexes where it also leads to TGFβ activation, leading to skeletal muscle fibroblast (FAP) activation and continued defective ECM generation that generates a reinforcing disease circuit that furthers the disconnection of the myofiber from the overriding basement membrane and ECM. To dive deeply into the molecular mechanisms of Ullrich CMD we propose 3 specific aims: Aim #1, to genetically increase/decrease myofibroblast activity in skeletal muscle of Ullrich CMD mice. Aim #2, to examine if loss of collagen VI leads to altered myofiber - ECM coupling in Ullrich CMD. Aim #3, to examine TGFβ as a mediator of myofibroblast-ECM-dependent disease in Ulrich CMD. A strength of this application is the use of genetically modified mouse models to directly assess the effects of the myofibroblast, TGFβ and myofiber-basement membrane detachment as primary disease initiating components of CMD in vivo, together generating a 3-way reinforcing feedback circuit. Finally, this work will suggest obvious translational approaches with gene therapy or biologics for treating Ullrich CMD (and related CMDs) based on the most proximal disease initiating mechanisms. For example, Aim #1 would suggest the use of reprogrammed CAR T-cells against activated fibroblasts, which was recently shown to antagonize fibrotic heart disease. Aim #2 would suggest gene therapy with a version of the Thbs4 cDNA that broadly upregulates integrins and the DGC to strengthen myofiber connectivity to the basement membrane broadly. Aim #3 would suggest the use of a selective TGFβ1 antagonizing monoclonal antibody (mAb) in patients.

NIH Research Projects · FY 2025 · 2017-09

PROJECT SUMMARY/ABSTRACT Sepsis is a leading cause of death in critically ill children and survivors of sepsis can have long-term problems. Obesity is a significant health problem but there is a benefit of obesity during critical illness, termed the obesity paradox. The mechanisms leading to the obesity paradox remain unknown. There are classically two functionally and histologically distinct types of adipose tissue: white adipose tissue (WAT) and brown adipose tissue (BAT). But a third type called beige adipocytes, also known as brown-in-white (brite) adipocytes, develop in response to various stimuli through a process known as WAT browning. Browning increases energy expenditure and oxygen consumption and may be detrimental during critical illness. The effect of obesity on the adipose tissue response to sepsis-induced critical illness has not been well explored in patients. Data from our laboratory demonstrates sepsis induces WAT browning and the sepsis-induced WAT browning is associated with increased inflammation and angiogenesis in non-obese but not obese mice. What is not known is whether adipose tissue remodeling occurs in children with sepsis and the impact of obesity. Based on these important data we are now able to translate these findings from murine sepsis to human sepsis. The long-term goal of our studies is to understand the mechanisms through which body fat, in normal and in excess, contributes to adipose tissue remodeling and affects outcomes in critically ill patients with sepsis. The central hypothesis of our proposal is sepsis-induced inflammation leads to adipose tissue alterations that impair recovery in non-obese patients. We plan to test our hypothesis and accomplish the objectives by completing the following three specific aims. In Aim 1 we will determine changes in body composition in critically ill children with sepsis. Patients will undergo noninvasive tracking of body composition with InBody Body Water Analyzer on admission and serially during their PICU course. In Aim 2 we will determine the mechanisms by which IL6 and STAT3 affect adipose tissue browning and the impact of obesity using ex vivo adipose spheroid sepsis models. In Aim 3 we will determine the role of angiogenesis and the impact of obesity using ex vivo adipose spheroid sepsis models. Aims 2 and 3 will utilize human adipose tissue obtained from obese and non- obese children undergoing abdominal surgery. We will identify the effect of sepsis on body fat in critically ill children and the mechanisms of adipose tissue remodeling. These findings may provide insight for future treatment strategies for critically ill patients.

NIH Research Projects · FY 2024 · 2017-09

ABSTRACT: Craniofacial anomalies (CFAs) comprise 75% of congenital defects and represent a biomedical burden of almost 700 million dollars per year in the United States. Surgical repair of CFAs is difficult and often requires a large source of skeletal tissue to replace/reconstruct the facial skeleton. Bone grafts used to repair CFAs frequently fail to integrate and are commonly allografts from mesodermally derived bone. This is a suboptimal tissue source since the facial skeleton is embryonically derived from an entirely different population of cells called neural crest cells (NCCs). NCCs; however, have not been used in tissue engineering approaches because a robust, postnatal source of cells does not exist and their multipotent nature raises concerns of regarding uncontrolled differentiation. The over-arching, long-term goal of my laboratory is to integrate our understanding of the cellular, molecular and biochemical mechanisms of NCC development and apply this knowledge towards generating novel therapeutic strategies for generating a robust source of NCC-derived tissues amenable for the surgical repair of craniofacial anomalies. To achieve this goal, we are focusing on precisely directing NCCs proliferation and differentiation into skeletal tissue via manipulation of the primary cilia, the cellular organelle which functions as the signaling hub of all cells. Gaining a firm understanding of how the primary cilia work to transduce molecular signals in NCCs, and other cells, will likely identify several novel therapeutic options for disease treatment. The impact of our work would be broad and far-reaching as it has the potential to revolutionize how CFAs and ciliopathies are treated.

NIH Research Projects · FY 2025 · 2017-08

OVERALL | PROJECT SUMMARY The goal of the CLEAR Consortium is to elucidate the developmental and genetic mechanisms of trachea- esophageal birth defects (TEDs) to better understand their etiology, enhance diagnosis, improve treatment, and inform strategies to generate tissue in vitro that might ultimately be used for repair. The trachea and esophagus arise from the separation of a common foregut tube during early fetal development. Defects in trachea- esophageal development cause a spectrum of life-threatening TEDs, which occur in ~1:3500 births and prevent proper breathing and feeding in newborn infants. Gene mutations are known to cause TEDs but have only been identified in ~15% of cases and how these cause congenital malformations is poorly defined. To address this unmet need we have assembled an experienced and highly collaborative multi-disciplinary team of clinicians, surgeons, geneticists, computational scientists, and developmental and stem cell biologists that use an innovative combination of patient genome sequencing, neonatal MRI, animal models, quantitative cell biology, single cell genomics, CRISPR gene editing and human PSCs-derived organoids to study TEDs. This Multi-PI project centered at Cincinnati Children’s Hospital (CCHMC) and Columbia University Medical Center (CUMC) is led by Wendy Chung MD PhD (CUMC), Paul Kingma MD PhD (CCHMC), Yufeng Shen PhD (CUMC), James Wells PhD (CCHMC) and Aaron Zorn PhD (contact PI; CCHMC). Our program has 3 projects linked together by an Integrated Genomics Core. Project-1: Comprehensive phenotypic and genetic assessment of TED patients. Project-2: Defining the developmental mechanisms of TEDs in animal models. Project-3: Modeling EA in human PSC-derived embryonic tissues.

NIH Research Projects · FY 2025 · 2017-08

ABSTRACT Sickle cell anemia (SCA) is among the world’s most common inherited blood disorders, and causes severe morbidity and early mortality. SCA is highly prevalent in sub-Saharan Africa, affecting over 300,000 births annually, with an estimated 30% increase in the next generation. To address the burden of SCA within Africa, neonatal screening is needed to establish the proper diagnosis, and hydroxyurea treatment is needed to ameliorate morbidity and decrease mortality. Hydroxyurea is listed by the World Health Organization as an Essential Medicine for children with SCA, representing the only realistic and affordable disease-modifying therapy in this setting. Until recently, hydroxyurea had been studied primarily in high-income countries, with virtually no data available regarding its safe and effective use in Africa. To address this critical unmet need, we designed and launched REACH (Realizing Effectiveness Across Continents with Hydroxyurea, NCT01966731), a prospective open-label study of hydroxyurea for young children with SCA in sub-Saharan Africa. In the current funding period, 606 children in four African countries received hydroxyurea escalated to maximum tolerated dose (MTD). Despite COVID, our research teams in Angola, Democratic Republic of Congo, Kenya, and Uganda collected unprecedented data on the safety, feasibility, and benefits of hydroxyurea for SCA in Africa, with >3000 patient-years of treatment. We documented reductions in sickle- related clinical events and found unexpected reductions in malaria, transfusions, and death. We performed whole exome sequencing to investigate inter-patient variability including hydroxyurea pharmacokinetics, pharmacodynamics, and pharmacogenomics. In the renewal, we will make additional contributions by extending hydroxyurea treatment to this unique cohort, whose average age is now 11 years and soon entering puberty, using a continued supply of hydroxyurea donated by Bristol Myers Squibb. Though our initial results are encouraging, REACH does not have a placebo-controlled cohort for comparison. Accordingly, we will enroll a new cohort of age-matched children with SCA at all four sites, to provide pre-treatment data for comparison to our treated cohort. In the first specific aim, we will assess the long-term effects of hydroxyurea at MTD to ameliorate SCA-related clinical complications and preserve organ function (especially brain but also kidneys, spleen, and eyes). We will obtain longitudinal data on the effects of hydroxyurea at MTD on physical growth, sexual development, and overall reproductive health, and collect serial DNA to test for the emergence of clonal hematopoiesis. In the second aim, we will investigate mechanisms by which hydroxyurea reduces malaria infections, combining epidemiological data with in vitro parasite invasion assays and an agnostic search for protective genetic polymorphisms. In the third aim, we will simplify and optimize hydroxyurea treatment using novel and innovative approaches, by testing the feasibility and safety of a pharmacokinetics-based dosing algorithm in the new patient cohort to minimize dose adjustments and lab monitoring, and then by validating our newly identified genetic polymorphisms that predict HbF treatment responses. REACH will expand hydroxyurea treatment in Africa, build local capacity, and establish a robust research infrastructure for future collaborations, including planned NIH curative therapies. REACH is uniquely poised to elucidate benefits and risks of extended hydroxyurea, allowing safe and evidence-based dosing in Africa.

NIH Research Projects · FY 2026 · 2017-07

Project Summary/Abstract Congenital heart defects (CHDs) are the most common congenital malformations. However, the molecular etiology underlying most CHDs remain poorly understood. Furthermore, CHDs even following surgery can lead to complications later in life that result in arrhythmias, stroke, and premature death. In order to develop novel therapies able to prevent CHDs and target therapies to specific cardiovascular tissues, it is critical to garner understanding of fundamental mechanisms directing normal cardiac chamber development and regeneration. Therefore, long-term goals of our lab are to understand conserved mechanisms that direct the development of individual cardiac chambers and chamber-specific mechanisms utilized during regeneration in vertebrates. Few signals are known to be required that specifically direct atrial development, with specific regulators of atrial regeneration not being understood. The specific aims of this proposal are to elucidate the mechanisms by which a syntenic long non-coding RNA (lncRNA) family limits the expression of Nr2f transcription factors and decipher how Nr2f protein levels affect atrial heterogeneity during development and atrial regeneration in adult zebrafish. The studies in this proposal are relevant to human health as numerous genomic analyses now indicate that mutations in Nr2f2 are associated with CHDs, in particular ASDs in humans. While Nr2f2 knockout mice and in vitro studies with human stem cells have revealed requirements for both Nr2f1 and Nr2f2 in atrial development, the mechanisms by which Nr2f proteins direct proper atrial development are not completely understood. Importantly, there is currently no understanding of lncRNA-dependent mechanisms regulating Nr2f proteins. Our analysis of a lncRNA we call as-oca shows that in vivo it represses the translation of nr2f1a, the functional equivalent of mammalian Nr2f2. Moreover, we find that Nr2f1a levels regulate previously unrecognized heterogeneity of atrial cardiomyocytes in the embryonic atrium and atrial regeneration. In Aim 1, we will examine the specific mechanism that as-oca inhibits nr2f1a translation and the conservation of this mechanism among the NR2F-associated lncRNA family in human induced pluripotent stem cells. In Aim 2, we will examine the requirements of Nr2f1a and canonical Wnt signaling in generating atrial cardiomyocyte diversity and the transcriptional signature of a previously unrecognized atrial subpopulation. In Aim 3, we will examine the requirement of the epicardium in atrial regeneration and requirement of Nr2f1a within the atrial epicardium. Because Nr2f transcription factors play conserved roles in atrial development of all vertebrates, these studies will dramatically improve our understanding of post-transcriptional mechanisms regulating normal vertebrate atrial development and unique mechanisms employed during atrial regeneration. Ultimately, these studies will garner a foundation of knowledge that can be used to improve therapies capable of preventing and ameliorating CHDs and efficiently repairing injured hearts.

NIH Research Projects · FY 2026 · 2017-04

Multiple sclerosis (MS) is a devastating disease of the central nervous system that affects over 2.3 million people worldwide. Current therapies for MS are at best only partially effective, and there is no cure. Improved understanding of MS disease mechanisms would lead to better diagnosis, treatment, and prevention. MS is caused by both genetic and environmental risk factors. Epstein-Barr virus (EBV) is the most consistently replicated environmental factor. Mounting evidence indicates that EBV-infected B cells, and their downstream immunological effects, are key drivers of MS disease processes. Recent work by our group and others implicates the EBV-encoded EBNA2 gene regulatory protein in mechanisms at almost half of the established MS genetic risk loci. In this proposal, we will test the hypothesis that EBNA2-driven allele-dependent alterations to human gene expression lead to distinct B and T cell phenotypes directly contributing to disease processes in MS. Aim 1. Quantification of EBV-specific human gene expression in multiple MS patient demographics. We will expand our cohort to include males and females of European, African, and Asian ancestries. We will measure MS- and EBV-specific human gene expression in these cohorts. Aim 2. Global discovery of EBV- and MS genotype-dependent gene regulatory mechanisms. We will use Massively Parallel Reporter Assays (MPRAs) to systematically identify EBV- and risk allele-dependent gene expression at all MS risk variants in primary B cells with and without EBV infection. We will identify human transcriptional regulators acting with EBNA2, EBNA3C, and/or EBNA-LP at these loci and confirm their allele- dependent actions in MS-derived B cells using cutting-edge functional genomics technologies. We will validate these EBV- and genotype-dependent gene regulatory mechanisms using CRISPR-based genome editing of patient-derived primary B cells at CD37, CD58, ZMIZ1, and other MS risk loci. Aim 3. Discovery of allelic EBV-based mechanisms altering cellular behavior in MS. We will gauge the necessity and sufficiency of EBV and specific MS variants (e.g: CD37, CD58, ZMIZ1) on B cell function using the CRISPR-edited B cells. We will measure impact on B cell receptor signaling, cytokine production, proliferation, and apoptosis. We will use an inducible pluripotent stem cell-derived blood brain barrier (BBB) endothelium model to measure the impact of EBV and MS risk alleles on B cell - BBB interactions. The concept that disease processes might be influenced by virus-controlled, allelic regulatory protein complexes is highly innovative and has never before been demonstrated. Our work for the first time provides mechanistic insight into the established role of EBV in MS through a unified gene by environment model. Comprehensive cataloging, dissection, and understanding of the downstream effects of genetic mechanisms impacted by EBV will be significant because it will provide strong rationale to develop therapies that interfere with these mechanisms, or even vaccines that prevent EBV infection to cure MS and other EBV-related diseases.

NIH Research Projects · FY 2026 · 2016-12

Project Summary/Abstract Induction of inflammation by canonical microbial ligands by engaging classical patten recognition receptors (PRR) has many beneficial outcomes including elimination of the pathogen and activation of adaptive immunity that serves as protection against reinfection. However, unwarranted inflammation can also be induced by aberrant activation of PRRs by noxious agents (toxins, uric acid etc) or because of naturally occurring mutations in sensors or adapters of the innate immune system leading to auto-inflammatory diseases such as Cryopyrin Associated Inflammatory Syndromes, and Interferonopathies. Auto-immune diseases on the other hand are different than auto-inflammatory diseases as the culprits that trigger pathology are self-reactive T and B cells. Auto-immune inflammation can lead to debilitating outcomes because of damage to vital organs such as kidney, pancreas intestines as well as Skin and joints. Paradoxically, many of the clinical treatments for T cell auto-immune diseases are all directed towards inflammatory cytokines made by the innate immune system. Our previous work demonstrated that effector and effector memory CD4 T cells have the capacity to drive IL-1b production, completely independent of pattern recognition receptor activation. We discovered that Effector CD4 T cells provide both signal 1 (TNFa) and signal 2 (FasL) to instruct the myeloid cells to produce IL-1b in a Caspase-8 dependent manner. The current proposal is based on very strong preliminary data that demonstrates that effector CD4 T cells in fact have the capacity to mimic microbial ligands to drive a broad pro-inflammatory program in cells of the innate immune system. We find that effector memory CD4 T cells induce additional genes in Dendritic cells that sets up important questions related to “T cell instruction” of the innate immune system. Here we posit that while proximal activation of PRRs is necessary for naïve T cell activation, effector memory T cells have the ability to directly activate the innate immune system thus bypassing the need for PRR sensing. Although this might have evolved as a beneficial arm of the innate adaptive cross-talk, we propose to understand the detrimental outcomes of adaptive instruction of innate immunity in driving inflammation and tissue pathology. In order to gain mechanistic understanding of innate inflammation driven by effector CD4 T cells, we propose three aims where 1. We will examine and characterize the nature of innate inflammation driven by different effector memory T cell lineages and identify the molecular players involved in this process, 2. We will investigate the molecular mechanisms by which effector memory CD4 T cells drive innate inflammation with a particular focus on STING and DNA damage and 3. We will examine the impact of CD4 T cell effector/effector memory CD4 T cell driven innate inflammation on auto-immune disease and pathology. Successful completion of these aims will provide novel insights into T cell driven innate inflammation independent and will open up new targets to treat auto-immune diseases.

NIH Research Projects · FY 2025 · 2016-09

Project Summary Motor vehicle crashes (MVC) are the leading causes of death among teens with eight teens dying per day in an MVC. Teens with Attention-Deficit Hyperactivity Disorder (ADHD) are at twice the risk of MVC compared to teen drivers without ADHD. A programmatic line of research by this investigative team has identified long (>2 secs) glances away from the roadway, particularly during engagement with secondary tasks, as being a key mechanism in ADHD teen driving risk. In the original research grant, the investigative team developed and tested a driver training program, enhanced FOcused Concentration and Attention Learning (FOCAL+), to specifically target reducing rates of extended glances away from the roadway in teens with ADHD. In a randomized controlled trial (RCT), teens with ADHD randomly assigned to FOCAL+ demonstrated 41% fewer long-glances and less variability in lane position during simulated driving assessments conducted immediately after the final training session, and 1- and 6-months post-training compared to teens assigned to modified driver's training. Moreover, during naturalistic driving over the course of a year of driving, FOCAL+ teens had 40% less risk of a crash/near-crash event than control teens. However, there are considerable barriers to disseminating FOCAL+ in its current format. FOCAL+, as implemented in the RCT, requires costly (~$90K) hardware and software that are quite complex to use. Though there has been much interest in offering this training since publication of our RCT results, key stakeholders have reported that the expense and complexity of the hardware and software requirements are barriers to adoption. In the proposed study, with input from relevant stakeholders, FOCAL+ training will be converted to an immersive virtual reality (iVR) platform. iVR- FOCAL+ will provide an affordable ($5K), single hardware, single software, easily-executable solution that implementation sites (i.e., driving schools, outpatient occupational therapy) will be able to afford, adopt, and offer to teens with ADHD. Using a Type 2 hybrid effectiveness-implementation design, teens with ADHD will be randomly assigned to receive either iVR-FOCAL+, the original FOCAL+ or a sham control group. The iVR- FOCAL+ training will be implemented in real-world, non-research settings (i.e., driving schools, outpatient occupational therapy). At baseline and 1- and 6-months post-training, teens' driving skills will be assessed during driving simulation. Naturalistic driving will be assessed during the year after training using video event recorders installed in the teen's car. Training costs and implementation outcomes (e.g., barriers to implementation) for each training will be collected. Using these data, we will examine the relative effectiveness and cost-effectiveness of iVR-FOCAL+ intervention compared to FOCAL+ training. Finally, implementation of iVR-FOCAL+ will be tested and described. The proposed research has the potential to facilitate adoption and eventual dissemination of a training program that can prevent injuries and fatalities among a high-risk population of teens as well as among those who share their roadways.

NIH Research Projects · FY 2025 · 2016-09

Abstract Genetic mouse models have been a powerful system to delineate key mechanisms of cancer development. However, there are likely to be gene expression and regulation differences between mouse and man that affect the applicability of some findings in these models. Furthermore, the inbred strains that are used may come with additional genetic caveats due to loss of heterozygosity. For these and other reasons, many cancer studies that identify targets or mechanisms in mouse systems benefit from replicating key findings in a relevant human cell model. Indeed, this has become a common expectation for high impact studies. Human pediatric leukemia is genetically diverse with many potential combinations of driver and accessory mutations, each potentially affecting phenotype. Therefore, a large bank of xenograft models is required to replicate the heterogenous nature of the disease as found in patients. Xenograft technology continues to advance rapidly. There has been a steady proliferation of immunodeficient mouse strains, each with specific strengths relative to each other. More sophisticated in vivo studies are now possible because of these new strains and many other technical advances in sample processing, detection, analysis, and modeling of therapeutic interventions. However, these mice are difficult to breed and manipulate due to their immunodeficient status. Human cells also require a different set of considerations. Human hematopoiesis in this environment is very different from what is found in mouse transplant systems. There are additional host versus graft and graft versus host issues to consider. Human cells require different culture conditions and transduction protocols. Human cells are morphologically distinct. The markers used for detection are very different, making flow cytometry experiments much more involved. I have spent the last 19 years working with these models and have made continuous improvements along the way. I have a well-documented history of developing new innovative methods and models for the leukemia research community. My ability to apply my unique experiences to the projects of many NCI funded investigators through the set of xenograft resources that I have built adds significant efficiency and value to the science that can be done at our institution. This proposal is aligned with NCI initiatives to use mouse models and genomics to identify new molecular mechanisms of carcinogenesis, delineate mechanisms of resistance, and identify new targets for drug discovery and development.

NIH Research Projects · FY 2025 · 2016-08

Updated Overall abstract: The central goal of the Cincinnati Rheumatic Disease Resource Center (CRDRC) is to promote studies that advance the understanding of pediatric rheumatic diseases and lead to new therapies for these diseases. The Cincinnati Rheumatic Disease Resource Center has two specific aims. Aim 1: Provide resources that will enhance the scope and breadth of the research community to advance the central focus of understanding inflammation and biological mechanisms contributing to the development of rheumatic disease in children and adults. Aim 2: Foster collaborations and interdisciplinary approaches to promote laboratory discoveries and generate translational research opportunities that lead to important patient-oriented outcomes. The Cincinnati Rheumatic Disease Resource Center includes an Administrative Core and four Resource Cores: Pediatric Rheumatology Tissue Repository (PRTR) Leader, Grant Schulert, MD, PhD Integrative Cell Phenotyping Core (ICPC). Leader, Sherry Thornton, PhD Functional Genomics Core (FGC). Leader, Leah Kottyan, PhD Bioinformatic and Modeling Core (BAM). Leader, Matthew Weirauch, PhD Collectively, these resources form a powerful infrastructure that fosters development of precision and predictive medical approaches based on genomics and disease mechanisms. The CRDRC will support disease-based research across the continuum of discovery, where laboratory findings generate translational studies that lead to clinical trials. In addition to advancing knowledge of pediatric rheumatic disease, the goals of the CRDRC include recruitment of established investigators to bring new expertise to the field, cultivation of collaborations within the local and national research community, and encouragement of young investigators committed to pursuing research careers focused on pediatric rheumatic disease. These goals of the CRDRC are particularly well supported by a Pilot Study Program that includes funding of work within the P30 cores and extends to institutional cores to broaden impact and support new areas of investigation. The CRDRC also will strengthen the research community through an enrichment program of local seminars, workshops, and symposia. A highly accomplished and collaborative community of researchers is already in place with expectations of major growth through new recruitment. An innovative Visiting Scholars program will aid in the dissemination of innovative approaches and the enrichment of faculty and trainees outside of Cincinnati. Together, these attributes create fertile ground for accomplishing the goals of the CRDRC, and ultimately to accelerate research to benefit pediatric rheumatic disease patients in their care. Updated Administrative core abstract: The Administrative Core will serve as the executive, coordinating and oversight component of the Cincinnati Rheumatic Diseases Resource Center (CRDRC). The Admin Core will provide executive, administrative (including fiscal), and personnel services to the CRDRC, as well as scientific oversight of the resource cores. This includes coordination of resources within the Research Cores to enhance ongoing studies and promote new projects. The Administrative Core will monitor progress and evaluate all aspects of the CRDRC’s operations, including appropriate expenditures by the Resource Cores and Enrichment Program. The Administrative Core will prepare required progress reports and regulatory documentation. Leah Kottyan, Ph.D., is the proposed Director of the CRDRC, and Alexi Grom, M.D. is proposed as Associate Director. Dr. Grom will function as Medical Director of the CRDRC by providing the liaison to the rheumatology clinic and the fellowship training program. The CRDCC’s chief executive and administrative body will be a four-member Executive Committee, which consists of the Director, Associate Director, and leaders of each Research Core. An Advisory Committee will provide advice to the Director and Executive Committee and will include local and external leaders with expertise to provide meaningful guidance to CRDRC leadership and Research Core directors. Members will include individuals experienced in core and center administration who are independent of the CRDRC, but who are part of, and familiar with, the academic setting within which the CRDRC resides or offer experiences of other comparable academic centers. Members will include individuals who complement the expertise of and utilize the Research Cores, and local leaders of programs that synergistically interact with the CRDRC. An Enrichment Program will be offered that includes research seminars, education relevant to technologies of the resource cores, and strategic planning of research goals. A novel and innovative P&F Program is proposed and will be managed by the Administrative Core to extend impact of Center resources to a larger number of investigators. This program will focus on advancing utilization of CRDRC cores or institutional cores to provide access to key technologies that might otherwise be cost-prohibitive. Projects will be solicited from the research community and the larger academic health center. A P&F Study Committee will assist in the selection of new P&F Studies and monitor progress of ongoing P&F Studies. This program will provide a valuable mechanism for advancing new investigators and new projects in the field of pediatric rheumatology. A Visiting Scholars program will fund travel grants to bring trainees and faculty to Cincinnati to learn from our Resource Cores. These grants are intended to provide opportunity and training for future research trainees, physicians, and faculty in the Research Community. In summary, the Administrative Core will enable the success and create fertile ground for accomplishing the overall goals of the CRDRC.

NIH Research Projects · FY 2025 · 2016-07

Principal Investigator/Program Director (Last, first, middle): Wu, Jianqiang Project summary Using technical language, briefly describe the research design and rationale for achieving the stated goals Neurofibromatosis type 1 (NF1) is an inherited disease predisposing affected individuals to benign Schwann cell tumors called plexiform neurofibromas (PNFs). Currently, prevention of PNFs is not possible, partly because the molecular mechanisms of tumorigenesis are not fully understood. Surgery remains the mainstay of therapy for PNFs. The FDA approved cytostatic MEK inhibitor, Selumetinib (Koselugo), shrinks tumor in 70% of individuals but tumors regrow after stopping drug treatment. Therefore, new therapeutic strategies and targets for the treatment of neurofibroma are urgently needed. The endoplasmic reticulum (ER) stress response pathways play pivotal roles in tumor growth and therapy in several cancers but remain unstudied in neurofibroma. Targeting these ER stress pathways might provide a novel therapy for PNF patients. Our new preliminary data show that: a) All three ER stress signaling pathways are activated in both mouse and human PNFs compared to controls. b) Knock down of protein kinase RNA-like endoplasmic reticulum kinase (PERK) by shRNA decreases neurofibroma like tumor number in a cell transplantation model in nude mice. c) Pharmacological inhibition of valosin-containing protein (VCP) together with a MEK inhibitor (MEKi) decreases cell proliferation, increases cell apoptosis and induces protein ubiquitination. Our central hypothesis is that loss of Nf1 in SC/SCPs leads to PNF formation by driving Runx- and VCP-regulated proteostasis to adapt to ER stress signaling, so that targeting proteostasis provides cytotoxic therapy for PNF patients. Two specific aims are proposed: In aim 1, we will determine if and how Runx and VCP regulate protein synthesis and degradation to maintain proteostasis so that Nf1-/- SC/SCPs adapt to ER stress, thereby driving PNF initiation and growth. In aim 2, we will test whether overwhelming irresolvable ER stress by targeting VCP (alone or in combination with MEKi) provides cytotoxic and, thus, durable control of PNF growth, and determine the mechanism of action. Overall, this proposal will provide mechanistic evidence of Runx1/3 and possible VCP- dependent proteostasis and adaptive ER stress signaling functions as oncogene on PNF formation and provide pre-clinical rationale for MEK-independent clinical trials.

NIH Research Projects · FY 2026 · 2016-05

Project Summary/Abstract Children born very preterm (VPT; <32 weeks’ gestation) face a disproportionate burden of neurodevelopmental impairments (NDI). Impairments in the subdomains of mathematics, reading, and behavior are the most common NDI, affecting 25-50% of VPT children. These abilities are critical to school readiness and success at work. In the first cycle of this grant, we assembled the largest North American cohort of VPT infants and performed advanced brain MRI at term corrected age (CA) with longitudinal neurodevelopmental assessments. We demonstrated the clinical importance of diffuse white matter abnormality (DWMA) and multiple other novel MRI biomarkers to enhance prediction of short-term NDI at 2 or 3 years CA. During this second funding cycle, our unique cohort will reach the critical early school ages of 5 to 7 years, when higher-order cognitive abilities in math and reading, emotional self-regulation, and behavior become more apparent and accurately testable. The ability to accurately predict these functions several years earlier could enable early interventions. Assessment of the dynamic trajectories of NDI in VPT children and their modifiable environmental and biologic causes can also accelerate the development of targeted neuroprotective interventions during these critical early years when neuroplasticity is at its peak. The overall objectives of this proposal are to map the early childhood trajectory of brain structural and functional changes and develop early and accurate prognostic models of school readiness. Our rationale is that elucidation of the long-term impact of DWMA, identification of the trajectory and mediators of NDI changes, and robust prognostic models of school readiness will enable novel early intervention trials in targeted VPT infants. To achieve our goals, we will perform morphometric, diffusion, and functional MRI at age 5 and comprehensive developmental testing at 5 and 7 years CA in our cohort of VPT children and a matched group of term-control children. To advance this highly impactful area of research, we propose the following three specific aims: (1) Determine the impact of DWMA at term CA on brain development and functional impairments at age 5; (2) Model individual trajectories of NDI and identify the mediators of neurodevelopmental changes between 3 and 7 years CA; and (3) Develop early prognostic models of school readiness. For the first aim we will correlate objectively quantified DWMA volume at term CA with brain structural and functional connectivity derived from advanced MRI and with higher-order cognitive functions, both at 5 years CA. Under Aim 2, we will plot individual trajectories of neurodevelopmental scores and identify the mediators of trajectory change between 3 and 7 years CA. For the third aim, we will apply the rich collection of clinical, biologic, and neurodevelopmental data collected between birth and 3 years CA to predict our three measures of school readiness at 5 and 7 years CA. This study will provide U.S. population-based prevalence data about the long-term adverse neuroanatomic and functional consequences of VPT birth, identify mediators of NDI change, and facilitate early prediction of school readiness to enable targeted EI and novel randomized clinical trials for VPT children.

NIH Research Projects · FY 2026 · 2016-05

PROJECT SUMMARY Cystic fibrosis (CF) is a progressive disease affecting around 30,000 people in the US and is caused by a mutation in a gene affecting the CFTR protein that regulates mucus composition. In airways in the lung this leads to mucus stasis, infection, and remodeling that result in mucus plugs, poor ventilation, and progressive airway destruction. Highly-effective CFTR modulators, now available to >90% of patients, have revolutionized clinical care, with increases in pulmonary function via more effective mucociliary clearance. Burdensome maintenance therapies like airway clearance treatment (ACT) require around 2 dedicated hours per day and have been questioned by patients, families, and medical providers. In a recent CF-community survey, airway clearance was ranked as the most burdensome therapy. Traditional studies of therapy withdrawal pose some patient risk, since measurement is via relatively insensitive pulmonary function testing (PFT) and lung-function reductions can have permanent consequences. Breakthroughs in structural and hyperpolarized-gas MRI demonstrate exquisite sensitivity to CF lung disease and can be used to monitor regional and subtle changes over time, much more precisely than PFTs, and with regional specificity. MRI provides a unique opportunity to safely evaluate ACT. The overarching goal of our proposal is to determine effectiveness of ACT in the era of highly effective CFTR modulators by studying structure-function relationships via MRI in patients of varying severity who have stopped and restarted ACT. We will achieve this goal via three separate, hypothesis-driven Aims in this clinical trial: Hypothesis 1: Patients who have relatively low structural defects will have fewer ventilation defects and higher pulmonary function, and these defects will relate to frequency of ACT usage. Specific Aim 1: To perform UTE and hyperpolarized Xe MRI in 30 CF patients aged 12-21, approximately 15 of whom have self-withdrawn ACT, to regionally characterize obstructive severity and correlate regional structural lung abnormalities (via UTE FLORET MRI) to functional deficits (via Xe MRI) Hypothesis 2: Patients who have self-withdrawn ACT after initiation of effective modulators will demonstrate increases in regional ventilation after reinitiating treatment, with greater ventilation increases in patients with a higher level of lung abnormalities. Specific Aim 2: To perform a stepwise ACT re-initiation trial in fifteen 12-21 y.o. patients who have self-withdrawn airway clearance treatment (defined as ≤ 3x/week). UTE and Xe MRI, spirometry, and multiple-breath washout will be performed at baseline, after increasing treatment to 7x/week for 1 week and then 14x/week for 2 weeks, with daily logging to aid compliance and study engagement. Hypothesis 3: Patients who currently use 2x daily ACT and have few lung abnormalities on MRI can reduce to 1x daily or less, with no significant functional changes in the lung. Specific Aim 3: To perform a stepwise ACT withdrawal trial in fifteen 12-21 y.o. patients who have low MRI abnormalities and high FEV1. Patients will be studied at baseline, after decreasing ACT to 7x/wk for 1 week, and after decreasing ACT to 3x/wk for 1 week.

NIH Research Projects · FY 2025 · 2016-05

This T32 application for the Research Training Program (RTP) for Pediatric Rheumatic Diseases seeks to equip pediatric rheumatology fellows with the investigative skills necessary to succeed in meaningful scientific careers in academic pediatric rheumatology. The program is based in a large Division of Pediatric Rheumatology (PR) with two P30 center grants and several NIH-funded investigators. The Pediatric Rheumatology Division, including its related clinic, is part of Cincinnati Children’s Hospital Medical Center (CCHMC) and The Cincinnati Children’s Research Foundation (CCRF), as well as part of the University of Cincinnati College of Medicine (UCCOM) with its clinical and biomedical science departments. The Pediatric Rheumatology divisional research program draws strength from uniquely integrated teams of clinical, translational, and basic researchers involved in active ongoing collaborations with members of Divisions of Immunobiology, Human Genetics, Hematology/Oncology, Bone Marrow Transplantation, Nephrology, Radiology, Psychology and Bioinformatics. The primary faculty advisors have been selected from 12 specialties divisions based on the ongoing collaboration with Rheumatology faculty, high research productivity and strong record of training of young scientists. In parallel to the two focus areas of divisional research, this T32 RTP has been offering two distinct research career pathways or training tracks. Trainees deciding to pursue laboratory based basic and translational research careers follow the Laboratory Research Track. Although most trainees in this track have been MD or MD/PhD, it also attracts PhD scholars who would like to apply their knowledge to address questions relevant to PR. The focus areas of the training in this track include basic disease biology and functional genomics in PRDs. Trainees deciding to pursue clinical research careers follow the Clinical Science Track. This track encompasses didactic and hands- on training in study design, biostatistics, research methods, clinical epidemiology, and outcome measure development. The program is well supported by core facilities as well as by academic courses, including a mandatory course in medical ethics and responsible conduct of research. Other available courses include computational system biology, immunobiology, molecular genetics, epidemiology, and biostatistics. The interdepartmental Immunobiology Graduate Program (MS and PhD) that is centered at CCHMC complements existing programs in Epidemiology and Biostatistics, and Education in the UCCOM. It is believed that this RTP provides unique resources and will continue to enlarge the pool of individuals with a career interest in biomedical science as it relates to pediatric rheumatology, a pool that is far too small to meet the requirements of the next decades as molecular medicine is increasingly applied in the clinic.

NIH Research Projects · FY 2026 · 2015-09

Project Summary This study's long-term goal is to elucidate the genetic and immunologic features of Eosinophilic Esophagitis (EoE). EoE is an emerging chronic disease that often starts in childhood and continues into adulthood and is associated with substantial morbidity, yet there are currently no FDA-approved therapies. Understanding this subject has significant implications as elucidating the fundamental genetic and immunologic features of the disease has potential to yield improved diagnostics and therapies. The central hypothesis of this proposal is that genome-wide association study (GWAS) interrogation, followed by genetic and biological validation, will uncover key processes involved in disease pathoetiology with a focus on the interface of adaptive and innate immunity. The rationale for this hypothesis is based on our prior studies, including initial GWAS that have identified disease susceptibility at chromosomes 2p23 and 5q22. Evidence is accumulating that the causal genes at these 2 loci are CAPN14 (calpain 14) and TSLP (thymic stromal lymphopoietin), respectively. These findings shift the focus from primary eosinophil defects to epithelial responses as being causal of EoE pathogenesis. Mechanistic studies have established that CAPN14 contributes to impaired epithelial barrier function and that TSLP promotes adaptive type 2 T cell immunity associated with overproduction of IL-5 and IL-13. CAPN14 sits at the interface of innate and adaptive immunity, as it is constitutively expressed by esophageal epithelium; however, it is also markedly induced by IL-13, likely derived from food antigen–activated Th2 cells. In addition to these 2 genetic loci (2p23, 5q22), GWAS have implicated numerous other suggestive loci, of which 11 have been recently preliminarily implicated using a custom-designed Illumina SNP array approach followed by preliminary functional analyses. Despite these advances, the causal gene variants and/or genomic pathways for EoE pathogenesis remain largely unclear. Herein, we will test the relevant and key hypothesis that GWAS interrogation, followed by genetic and biological validation, will uncover disease pathoetiology. We will test this central hypothesis via 3 complimentary aims using innovative approaches that combine genetic and biological studies. In Aim 1, we will focus on a primary GWAS lead, CAPN14. We will test the hypothesis that CAPN14 is an essential regulator of cellular junctions and barrier integrity and contributes to IL-13–induced, EoE-related epithelial responses. We will identify its binding partners and potential substrates and the consequences of CAPN14 deficiency in esophageal epithelial cells and CAPN14 transgenic overexpression in mice. In Aim 2, we will test the hypothesis that meta-analysis of additional EoE cohorts analyzed by GWAS will refine the involvement of implicated loci/genes and identify new variants. In Aim 3, we will interrogate disease-associated single-nucloeotide polymorphisms (SNPs) by a combination of innovative genetic, chromatin mapping, transcriptomic expression profiling, and functional biological studies to establish relevance of the genetic variants and identify causal genes.