Research Inst Nationwide Children'S Hosp

NIH Research Projects · FY 2024 · 2020-06

ABSTRACT Congenital urinary tract obstruction (UTO) is the leading cause of chronic kidney disease in children. There is a critical need for measures to prevent obstructive nephropathy – the renal injury and dysfunction that result from UTO. The urothelial lining of the kidney undergoes massive reorganization in response to congenital and acquired UTO, but until recently the significance of this urothelial remodeling has remained unclear. Recently published data from our group provide the first experimental evidence that renal urothelial remodeling limits the extent of obstructive nephropathy. During obstruction, the renal urothelium protects the kidney from injury by producing an apical plaque composed of Uroplakin proteins. Depletion of the urothelial plaque accelerates parenchymal loss in a mouse model of congenital UTO, accompanied by renal failure and death. Urothelial plaque depletion likewise results in increased parenchymal loss following unilateral ureteral obstruction, a model of acquired UTO. These data in congenital and acquired UTO models provide strong support for our central hypothesis that the urothelium plays an essential role in protecting the kidney from obstructive nephropathy. The objectives of this application are three-fold. First, we aim to define the role of urothelial injury as a driver of obstructive nephropathy. Second, we will test the hypothesis that pharmaceutical enhancers of Uroplakin+ cell generation and Uroplakin expression can be harnessed therapeutically to limit obstructive nephropathy. Last, we will test the potential of urothelial proteins as diagnostic biomarkers of obstructive nephropathy in mice and in children undergoing evaluation for congenital UTO. The long-term objective of this project is to improve the care of patients with UTO by identifying novel diagnostic, prognostic, and therapeutic approaches to prevent progressive chronic kidney disease.

NIH Research Projects · FY 2025 · 2020-05

Abstract Ewing sarcoma (EwS) is a pediatric and young adult cancer that is driven by the EWSR1-FLI1 translocation. Despite decades of work, this cancer is still an enigma, with poorly understood biology and no targeted treatments. Our recent work published in Nature demonstrated a previously overlooked consequence of EWSR1-FLI1, that this fusion causes hyperphosphorylated RNA polymerase II (pRNAPII) due to loss of EWSR1 inhibition of CDK7 and CDK9. We observed high levels of transcription, with high levels of R-loops present in locations that R-loops normally (physiologically) occur. Based upon these findings, we began to reconsider cellular phenotypes of EwS to identify the molecular basis of these phenotypes and ask whether these changes provide a fundamental defect in all EwS. One phenotype that was previously identified in EwS is that these cells display altered splicing profiles. In recent years there were several reports linking R-loops to splicing, with splicing defects causing R-loop accumulation and R-loops being associated with sites of alternative splicing. Further, it was reported that the splicing machinery inhibits DHX9 (aka RNA helicase A; RHA) from causing accumulation of toxic R-loops. Also, of interest, is that EWSR1-FLI1 interacts with and impairs DHX9 activity. By performing a genomic RNAi viability screen, we determined that EwS is acutely sensitive to a loss of RNA processing capability. These collective observations led us to the hypothesis that Ewing sarcoma is dependent upon RNA processing machinery to prevent accumulation of toxic R- loops. If our hypothesis is correct, then it suggests that there may be a therapeutic opportunity to target splicing components, converting the high levels of physiological R-loops in EwS into pathological R-loops to drive toxic genomic instability. We propose to test our hypothesis with two Aims. In Aim 1, we will investigate the mechanistic relationship between transcription levels, R-loops and splicing in EwS. For this we will modulate splicing components by siRNA depletions, cDNA expression or use of pharmaceutical inhibitors, examining transcription activity (Gro-Seq and uridine incorporation), splicing (reporters and RNA-Seq analysis) and R-loops (DRIP-Seq). In Aim 2, we will examine whether EwS is particularly reliant on splicing components or RNA:DNA helicases to block toxic conversion of R-loops and how targeting these processes impacts EwS viability, DNA damage response and/or cell cycle progression. We will ask if these modulations effect EwS cells at a particular time during cell cycle or stem cell state using single cell sequencing techniques. We will also assess how these various components of R-loop biology interact with one another, with pRNAPII and with R-loops in EwS. Finally, based upon these results, we will extend our findings to test efficacy of removing the R-loop metabolizing program that EwS is most reliant upon as a means to inhibit EwS tumor growth. Overall, this work should provide critical insight into the biology of Ewing sarcoma and provide new avenues for treatment beyond the standard chemotherapeutics currently used.

NIH Research Projects · FY 2025 · 2020-02

Project Summary: Natural Killer (NK) cells are lymphocytes of the innate immune system. NK cells defend us by inducing antibody-dependent cell mediated cytotoxicity (ADCC) where NK cells lyse antibody coated virally- infected target cells. Recent experiments showed generation of long-lived “memory-like” NK cells, similar to memory lymphocytes in the adaptive immune system, in mouse and humans challenged by viral infections (such as cytomegalovirus). These memory NK cells generated a more vigorous ADCC response compared to their naïve counterparts which make the memory NK cells an attractive candidate for augmenting monoclonal antibody based immunotherapies against cancer and infectious disease. However, two major issues limit the use of “memory-like” NK cells for such therapies: 1) A rudimentary understanding of mechanisms underlying NK cell-mediated ADCC is lacking; and 2) humans and mice show key differences in the NK cell signaling networks, which regulate ADCC. We address the above challenges by developing computational models with predictive powers for antibody responses induced by NK cell subsets (from naïve to memory) in humans and mice by synergistically combining data-driven and mechanistic in silico models (rooted in statistical physics, nonlinear dynamics, information theory, statistics, and chemical engineering) with single cell mass cytometry by time of flight (CyTOF) and state-of-the-art wet lab experiments in primary NK cells obtained from human subjects and genetically modified mice. The objective of the proposal is to quantitatively characterize mechanisms underlying ADCC in diverse NK cell subsets in humans and mice and then use this quantitative understanding to develop novel mouse models of ADCC that reflect the situation in humans more accurately. We will pursue two aims: Aim 1: Modeling ADCC activity in human naïve and memory NK cell subsets. Aim 2: Modeling ADCC in mouse NK cells. The expected outcome of quantitative characterization of the differences and synergies in mechanisms of ADCC induced by CD16 and CD32 receptors in different NK cell subsets (naïve to memory primary NK cells) (Aim 1) will help us generate improved mouse models that more accurately represent ADCC mediated by human NK cells (Aim 2). This unique framework will provide the scientific community with a mouse model for ADCC that more accurately reflects the situation in humans, a critical asset for pre-clinical development of monoclonal antibody therapeutics for cancer and infectious diseases.

NIH Research Projects · FY 2026 · 2019-04

PROJECT SUMMARY / ABSTRACT The Cooperative Human Tissue Network (CHTN) was established as a mechanism to provide the scientific community with high-quality human biospecimens for research. The CHTN allows investigators to access the biospecimens they need to perform cutting-edge research, including basic and early translational cancer research, and laboratory assay development. As one of six CHTN Divisions, the Pediatric Division of the CHTN (pCHTN) will receive, process, and distribute scientifically relevant biospecimens collected from children, adolescents, and young adults. These biospecimens can include malignant, benign, disease-involved, and uninvolved biospecimens. The pCHTN seeks to directly promote and support cutting-edge research in the diagnosis, biologic behavior, and treatment of acute and chronic diseases including cancer by pursuing the following specific aims: 1. To leverage the relationship with the Children’s Oncology Group , in addition to Nationwide Children’s Hospital Department of Pathology and Laboratory Medicine to procure a wide range of well-annotated biospecimens from a variety of both cancerous and non-cancerous conditions, including biospecimens from rare tumor types. 2. To support cutting-edge research by distributing high-quality biospecimens to approved investigators. 3. To maximize limited biospecimen resources and provide economies of scale by performing additional processing of biospecimens (e.g., nucleic acid extraction, tissue microarray creation, digital images of stained tissue slides, etc.) 4. To maintain a reliable and robust Quality Management Program that monitors all aspects of biospecimen collection, processing, storage, and distribution in an effort to continuously evaluate and improve our operational capabilities, assess best practices, ensure investigator satisfaction, and protect the privacy and confidentiality of those individuals from whom the biospecimens and data were obtained. 5. To actively contribute to the CHTN by participation and collaboration in Network interactions, particularly in fulfilling highly customized or non-standard requests, participating in collaborative Network activities, and coordinating or developing new strategies to ensure that the CHTN remains responsive to the needs of the scientific community.

NIH Research Projects · FY 2025 · 2018-01

PROJECT SUMMARY Tissue engineered vascular grafts (TEVGs) have demonstrated potential to revolutionize cardiovascular care, with multiple grafts now in clinical trials in children and adults. Yet, there remains a pressing need to optimize these grafts to improve outcomes and enable wide-spread usage. In this proposal, we build upon a strong foundation of prior findings but introduce an innovative multi-fidelity computational-experimental approach that promises to accelerate greatly the development of improved TEVGs. Although the proposed approach is general with broad applicability, we will focus on one particular application – TEVGs for congenital heart surgery – to refine the approach and illustrate its utility. Specifically, we will use a pre-clinical juvenile ovine model to collect the longitudinal data needed to develop and inform novel multiscale computational models that will be melded to describe the in vivo development of a neovessel from an implanted biodegradable polymeric scaffold. Our approach will be informed by data from three initial, non-optimal designs, then used to identify via formal methods of optimization preferred microstructural scaffold parameters and an overall geometry that optimizes in vivo function. Particularly novel will be our ability to account for normal developmental changes in the lamb vasculature and coupling of cell signaling, growth and remodeling, and 3D hemodynamics in a novel multi-fidelity, multiscale workflow that allows optimization of desired biological and physiological outcomes. To achieve these goals, we propose three Specific Aims: 1) To quantify normal vascular development and performance of three baseline TEVG designs in a lamb model; 2) To develop and employ a novel multiscale fluid-solid-growth (FSG) simulation framework to optimize TEVG design; 3) To validate the model-identified optimal TEVG design in a longitudinal large animal study. Our team is uniquely positioned for success, combining expertise in animal models of congenital heart disease, development of TEVGs and their clinical translation, finite element simulations of cardiovascular hemodynamics and biomechanics, modeling vascular growth and remodeling, and identifying and modeling mechanisms of mechanobiology. Our approach is innovative in that we will 1) meld macro (organ) level simulations of cardiovascular biomechanics with micro level simulations of vascular cell signaling, 2) develop a novel, generally applicable paradigm for model-driven optimization of tissue engineered structures that provides control over outcomes, and 3) facilitate clinical translation of TEVGs with improved performance. Successful completion of this study will be significant in multiple ways – not only will it result in a new (optimal) design of a TEVG for use in the Fontan surgical procedure, performed in children born with single ventricle congenital heart defects, it will also establish a novel computational-experimental paradigm in cardiovascular tissue engineering that promises to accelerate the development of diverse implants.

NIH Research Projects · FY 2026 · 2017-01

PROJECT SUMMARY Patients with lower extremity peripheral artery disease (PAD) are at increased risk of lifestyle limiting claudication, ulceration, amputation, and death. Despite this knowledge, an ongoing challenge for the vascular community is the lack of a standard non-invasive test for quantifying regional abnormalities in skeletal muscle perfusion in PAD. Our laboratory has been at the forefront of translating nuclear perfusion imaging techniques to patients with PAD and demonstrated the potential prognostic value of these methods for prediciting clinical outcomes. Specifically, in the prior funding period of this award, we clinically translated a single photon emission computed tomography (SPECT)/CT imaging approach from pigs with limb ischemia to patients with PAD and demonstrated that SPECT/CT perfusion imaging measures were repeatable in patients, detected regional abnormalities in resting perfusion of the lower extremities, assessed perfusion defects that corresponded with the level of future amputation, and quantified revascularization-induced regional improvements in relative perfusion that were associated with risk of amputation after endovascular therapy. We then developed and validated a dynamic positron emission tomography (PET) perfusion imaging approach using fluorine-18 (18F)-sodium fluoride (NaF), a radionuclide traditionally used for molecular imaging of bone and vascular microcalcification. Dynamic 18F-NaF PET imaging revealed utility for quantifying regional, absolute measures of muscle perfusion (i.e., ml/min/100g) in pigs and PAD patients with limb ischemia that coincided with microvascular remodeling in pig skeletal muscle and symptom severity in PAD patients. The goal of this proposal is to leverage the novel PET perfusion imaging method developed in the prior funding period to optimize regional evaluation of absolute measures of skeletal muscle perfusion in PAD patients undergoing endovascular revascularization. To accomplish this goal, in Aim 1, we will validate dynamic 18F- NaF PET perfusion imaging as an approach for monitoring ischemia-induced alterations in skeletal muscle morphology and mitochondrial function using a newly developed porcine model of chronic limb-threatening ischemia (CLTI). In Aim 2, we will establish a healthy clinical database for 18F-NaF PET perfusion measures and compare these perfusion measures to standard indices of macro- and microvascular disease in patients with PAD. In Aim 3, we will quantify absolute changes in regional calf muscle perfusion in response to revascularization procedures in patients with PAD using dynamic PET imaging, and evaluate the relationship between regional perfusion responses and clinical outcomes. These studies are designed to pre-clinically validate the utility of dynamic PET perfusion imaging for evaluating adverse remodeling of skeletal muscle and test the clinical efficacy of this method for monitoring regional responses to endovascular treatment and predicting clinical outcomes.

NIH Research Projects · FY 2025 · 2015-08

1 Summary: 2 CD4 T cell activation provides a model system for studying molecular mechanisms that coordinate a wide variety 3 of often competing physiological processes. When CD4 T cells encounter an antigen in the proper context, they 4 rapidly accumulate biomass, undergo extensive expansion, and differentiate into functional lineages that spe- 5 cialize on cytokine production. Robust and effective CD4 T cell-mediated immune responses require proper al- 6 location of metabolic resources through the central carbon metabolic pathways to sustain energetically costly 7 processes like growth, proliferation, and cytokine production. Also, ancillary metabolic pathways, such as amino 8 acid catabolism and polyamine (PA) biosynthesis, are critical to regulating T cell proliferation and inflammation. 9 The objective of this proposal is to understand how the arginine-polyamine metabolic axis is regulated during 10 T cell activation and ultimately contributes to inflammation and autoimmunity. Our lab recently revealed that the 11 transcription factor, c-Myc, controls an ancillary metabolic pathway that connects arginine (Arg) catabolism to 12 the biosynthesis of PAs, which are an essential class of polycationic metabolites ubiquitously present in all living 13 organisms. Unlike most other amino acids that are primarily used for anabolic protein synthesis during T cell 14 activation, most cellular Arg is catabolized and funneled into synthesizing PA (Arg-PA metabolic axis). Genetic 15 and pharmacological perturbation in the intracellular PA pool suppresses proliferation, suppresses TH1 and TH17 16 differentiation, but enhances iTreg differentiation. Hence, we hypothesize that the arginine-polyamine metabolic 17 axis orchestrates a metabolic checkpoint to optimize CD4 Teff cell proliferation and inflammatory re- 18 sponse. This checkpoint may be therapeutically exploited by polyamine blocking therapies. The aims of 19 this proposal are to 1) decipher the Arg-PA metabolic axis reprogramming and assess the impact of crucial 20 metabolic steps on Teff cells in the context of modulating Myc; 2) determine the outcomes of modulating Arg-PA 21 axis in regulating the effector function of T cells; 3) assess the contribution of PA de novo biosynthesis and the 22 PA salvage pathway to the intracellular PA-pool and T cell proliferation and effector function, and 4) develop and 23 test complementary enzymatic, genetic, and dietary strategies to exploit the Arg-PA axis to modulate inflamma- 24 tory response and autoimmunity in animal models of multiple sclerosis and rheumatoid arthritis. Collectively, the 25 expected outcomes of this project are significant as it will reveal the fundamental principles of the emerging 26 connections between cell metabolism, immune signaling, and T cell differentiation. These studies are critical to 27 developing novel approaches and therapeutic interventions that improve clinical outcomes of inflammatory and 28 autoimmune diseases.

NIH Research Projects · FY 2025 · 2015-06

Salmonellae are Enterobacteriaceae that cause a spectrum of diseases in humans and animals, including enteric (typhoid) fever and gastroenteritis. Typhoid fever caused primarily by Salmonella enterica serovar Typhi (S. Typhi), results in a life-threatening systemic disease that is responsible for significant morbidity and mortality annually worldwide. Approximately 5% of individuals infected with S. Typhi become chronic carriers with the gallbladder (GB) as the site of persistence. S. Typhi is a human- restricted pathogen, therefore asymptomatic carriers represent a critical reservoir for further spread of disease. We have demonstrated that gallstones (GSs) aid in the development and maintenance of GB carriage in a mouse model (utilizing S. Typhimurium, which causes a typhoid-fever like disease in mice) and in humans, serving as a substrate to which Salmonellae attach and form a protective biofilm. Thus, biofilm formation is a key step in the establishment of carriers. Traditional antibiotic therapies are usually incapable of clearing chronic S. Typhi infections, as the biofilm phenotype renders the bacteria tolerant to the mechanisms of these drugs. Thus, the identification of novel therapeutics capable of targeting S. Typhi biofilms is necessary in order to eliminate chronic carriage and eradicate the disease. Towards this end, our group has identified four small molecules and two antibodies capable of inhibiting and/or disrupting Salmonella biofilms in vitro. We advance two of the small molecules in this proposal, JG-1 and M4, that both inhibit and disrupt biofilms in vitro and reduce GB bacterial numbers in vivo. We hypothesize that the use of these anti-biofilm compounds in conjunction with an antibiotic will more effectively inhibit and disrupt Salmonella biofilms in vivo in our mouse model of chronic carriage when compared to the administration of antibiotic therapy alone. In Aim 1, we will assess the efficacy of these anti-biofilm compounds at preventing and treating chronic infection compared to traditional antibiotics alone by utilizing our established mouse model of typhoidal chronic carriage. We will also measure important pharmacokinetic and tolerability parameters of these compounds. In order to elucidate the mechanisms by which these compounds antagonize Salmonella biofilms, in Aim 2 we will identify the specific bacterial target(s) of each compound by enriching for mutants exhibiting compound resistance and by performing direct pull-downs of targets from bacterial lysates. Structure activity relationships and derivatives with enhanced physiochemical and biological properties will be generated in Aim 3. In summary, we propose an investigation into the safety, efficacy, and mechanisms of novel anti-biofilm compounds to prevent and treat chronic infections by typhoidal Salmonella. To our knowledge this study will be the first attempt (utilizing subject experts in anti-biofilm medicinal chemistry and Salmonella chronic infection) to specifically target Salmonella biofilm formation in vivo as a means of combating chronic carriage.

NIH Research Projects · FY 2026 · 2015-04

Project Summary SWOG is an adult clinical trials cancer group within the National Clinical Trials Network (NCTN) of the National Cancer Institute (NCI). SWOG clinical trials include a range of cancer types (e.g., breast, lung, colon, prostate, leukemia, melanoma, lymphoma, and multiple myeloma). The SWOG Biospecimen Bank (Biobank), located in the Biopathology Center (part of the Abigail Wexner Research Institute of Nationwide Children's Hospital), was designed to procure, process, bank, and distribute biospecimens collected as a major component of NCI- sponsored clinical trials. These biospecimens are linked to demographic information, surgical and pathological reports, and treatment and follow-up data of study participants enrolled on SWOG protocols. Upon review and approval, biospecimens are prioritized for distribution to the specific SWOG investigators defined in the clinical trial protocols. After the needs of the planned research have been addressed, residual (legacy) banked biospecimens can be distributed to other investigators within and outside SWOG via the NCTN Navigator. The SWOG Group and Biobank Concierges can also facilitate access to SWOG legacy biospecimens that are not currently uploaded in Navigator. This process has been designed to mirror the process for legacy biospecimens on the Navigator system and includes a feasibility assessment by the SWOG Biobank and the SWOG Statistical and Data Management Center prior to proposal submission. The Biobank is a functioning and viable part of SWOG's clinical and translational medicine program, and focuses on the integration of diagnostic, therapeutic, laboratory, and clinical data to answer questions in clinical and translational research. The SWOG Biobank works under the hypothesis that banking of high quality, clinically-annotated and cancer- related human biospecimens, linked to molecular signatures/markers, will significantly facilitate and further advance clinical cancer research. To support this effort, the SWOG Biobank manages these biospecimens under strict guidelines and standard operating procedures based on current Best Practices for biorepositories and the latest laboratory technologies. The Biobank's experience, coupled with a firm commitment to quality and integrity operations, makes it uniquely able to provide an accurate and effective resource of tumor and normal samples with associated clinical, epidemiologic, and protected health information. The SWOG Biobank aims to harmonize and expand general banking practices and innovative technologies, which will continue to facilitate molecular biology research and increase investigator participation in clinical research trials via the NCI-sponsored Group Banking Committee (GBC). The SWOG Biobank also plans to support the processing, banking, and distribution of biospecimens to the Cancer Immune Monitoring and Analysis Centers (CIMACs).

NIH Research Projects · FY 2026 · 2015-04

Project Summary The Children’s Oncology Group (COG) is the pediatric clinical trials group of the National Clinical Trials Network (NCTN). Over 90% of United States and Canadian children and adolescents with cancer are treated at COG institutions with no racial, ethnic or geographic bias to registration, allowing for near- population-based clinical, translational and basic research. COG has established a strong track record of collecting and banking tumor and non-diseased biospecimens from patients enrolled in COG-sponsored clinical trials, most of which include randomized treatment questions. The COG Biospecimen Bank (COG Biobank), located within the Biopathology Center (part of the Abigail Wexner Research Institute at Nationwide Children’s Hospital), aims to provide high-quality pediatric and adolescent human malignant biospecimens to the research community. Upon review and approval, biospecimens are prioritized for distribution to the COG investigators defined in the clinical trial protocols. After the needs of the planned research have been addressed, residual (legacy) banked biospecimens can be distributed to other investigators within and outside COG via the NCTN Navigator The COG Group and Biobank Concierges can also facilitate access to COG legacy biospecimens that are not currently uploaded in Navigator. For all biospecimen distribution projects, a feasibility assessment by the COG Biobank and the COG Statistical and Data Management Center (SDMC) is completed prior to proposal submission. This proposal supports the collection, processing, and distribution of biospecimens from patient enrolled on COG-sponsored trials. The Pediatric Division of the Cooperative Human Tissue Network (CHTN) then facilitates the distribution of the approved legacy biospecimens. The COG Biobank seeks to directly promote and support outstanding research in the diagnosis and treatment of pediatric cancer through centralized collection, quality control, storage, and distribution procedures. The COG Biobank will continue efforts to provide the latest biorepository-based technological innovations and best practices, resulting in constant improvements in our operational capabilities, investigator access to biospecimens, investigator satisfaction, and the stewardship of these precious resources. Under proper regulatory guidelines and in association with the COG SDMC, the COG Biobank will support translational aspects of cutting-edge research by ensuring appropriate tracking of biospecimens as well as linkage to existing demographic, clinical, biological, treatment, and outcome data. Finally, the COG Biobank will provide investigators with a comprehensive database solution and dynamic informatics tools that facilitate pediatric cancer research. The COG Biobank also plans to support the processing, banking, and distribution of biospecimens to the Cancer Immune Monitoring and Analysis Centers (CIMACs).

NIH Research Projects · FY 2025 · 2014-05

Project Summary Duchenne muscular dystrophy (DMD) is a degenerative muscle disorder that affects approximately 1:3500 to 1:5200 live male births caused by mutations in the X-linked DMD gene. DMD gene mutations result in absence of the dystrophin protein in muscle fibers, leading to myofiber necrosis, endomysial fibrosis, and fat replacement. It is a devastating disorder, leading to loss of ambulation by age 12, and historically to death by age 20. The psychological and socioeconomic effects on families are enormous; these include but are not limited to the costs of medical care, opportunity costs for career and work, and the psychological toll taken on parents and siblings. Our long-term goal is to understand which genes modify disease progression and severity of DMD. Confirming a hypothesis derived from a genetic modifier of muscular dystrophies in mice, we have recently used data from patients enrolled in the United Dystrophinopathy Project (UDP) to demonstrate that polymorphisms in the LTBP4 gene influence age at loss of ambulation. Our objective in this project is to identify additional genetic modifiers of skeletal muscle, cardiac, and ventilatory function, and our central hypothesis is that such modifiers can be identified by use of the UDP database, a unique resource that contains detailed phenotypic data and archived DNA samples from over 900 DMD patients. Our specific aims are to 1) update and analyze phenotypic data within the UDP cohort, 2) map modifier traits by high- density single nucleotide polymorphism arrays, and 3) validate newly identified putative genetic modifiers. Validation will engage collaborating networks of investigators in the US and Europe, who have additional natural history cohorts of DMD patients. At the conclusion of these Aims, we will have gained new information about modifier genes associated with the severity and progression of DMD.

NIH Research Projects · FY 2025 · 2013-09

Abstract Primary glomerular diseases, including minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), immunoglobulin A nephropathy (IgAN), and membranous nephropathy (MN), are associated with significant morbidity and mortality in both adults and children. Cure Glomerulonephropathy (CureGN) is a prospective, longitudinal observational cohort study launched in 2013 to address critical knowledge gaps in the pathogenesis, natural history, and response to therapy of these heterogeneous disorders. It is a study of unprecedented size and remarkable depth, built by a unique collaborative interdisciplinary community. The international consortium includes researchers with diverse expertise, affected patients and advocacy groups, the biopharmaceutical industry, and federal funding agencies. CureGN has successfully recruited a diverse cohort of nearly 2800 adult and pediatric participants with MCD, FSGS, IgAN and MN from more than 60 clinical study sites. Biospecimens, clinical data, and patient reported outcomes are collected to enable high- quality clinical, mechanistic, and translational investigations. This foundational work is being conducted by a collaborative infrastructure including the Data Coordinating Center (at the University of Michigan, Northwestern University and Cleveland Clinic) and four Participating Clinical Centers (managed at the University of Pennsylvania, Columbia University, University of North Carolina, and the Pediatric Nephrology Research Consortium). CureGN is paving the way for personalized care in glomerular disease by disentangling the heterogeneity within these disorders that are etiologically diverse but currently grouped into only four diagnoses. In CureGN’s third study phase, we propose to maintain and enhance the CureGN Consortium infrastructure and ancillary studies program to accelerate patient-relevant glomerular disease research. We will continue our core observational study, enrolling additional participants in a recruit-to-replace strategy to maintain an active cohort of 2000 participants with high quality clinical data and biospecimens. We will implement state of the art tools for remote data and biospecimen collection, expand biospecimen types, and implement the use of new mobile applications for patient engagement and medical record linkages. Mature scientific working groups, committees and ancillary infrastructure will continue to support a multidisciplinary core and ancillary study program to achieve the scientific goals of CureGN. We will continue our outreach to the scientific community by enhancing CureGN’s role as an outstanding training vehicle for the next generation of glomerular disease researchers and attracting cutting-edge, established scientists to glomerular disease through opportunity pool grants, collaborations with patient advocacy groups and professional societies, training workshops and support of ancillary studies from academic and industry partners. Through this coordinated effort, CureGN is prepared to accelerate improvements in the care of patients with glomerular disease.

NIH Research Projects · FY 2025 · 1999-09

PROJECT SUMMARY Despite the relative success of pneumococcal conjugate vaccines (PCVs) against pneumococcal otitis media (OM), the incidence of all-cause OM remains unacceptably high worldwide and broad use of PCVs has also changed the microbiology of OM to one in which nontypeable Haemophilus influenzae (NTHI) predominates. Additionally, whereas the importance of biofilms in OM pathogenesis, chronicity and recurrence is recognized, and the recalcitrance of biofilms to antimicrobials is well accepted, we nonetheless still treat OM with oral antibiotics that do not reach levels in the middle ear required to kill planktonic bacteria, let alone those resident in a biofilm. The use of broad-spectrum antibiotics is also not without consequence. Rashes and diarrhea are common, and we now know that early life use of antibiotics disrupts development of the very gut microbiome that is essential for normal human immune system development. Due to global concerns about these issues, in a recent Nature commentary, thought leaders encouraged us to consider new ways to combine the protection offered by effective induced antibodies with the more appropriate use of antibiotics as our “last hope against multi-drug resistant bacteria and persistent disease”(55). With the recent recognition that bacteria which cause persistent diseases exist in not only planktonic or biofilm-resident states, but also a third distinct newly released (NRel) state that is the most sensitive to killing by antibiotics, we envision use of specifically targeted antibodies to release bacteria from their highly resistant biofilms so they can now be killed by both host immune effectors and, if necessary, traditional antibiotics but now used at a markedly reduced dose and for a limited time. We demonstrate via many new preliminary data that NRel NTHI are highly unique and further, that NRel NTHI populations are distinct from each other, dependent upon the specific effector that induced their release. Here, we strive to combine our expertise in immunology with both our advanced understanding of biofilm structural biology and our growing appreciation of the NRel NTHI state. Integration of these concepts provides us with the opportunity to develop a truly novel approach that we believe could offer a viable option with which to combat chronic and recurrent OM. We will utilize our understanding of both the NRel phenotype and the activity of specifically directed antibodies to leverage their combined power. In Aim 1, we will use established in vitro assays to determine how environmental conditions (singly and in combination) affect the kinetics, extent of duration and phenotype of NRel bacteria as a target for intervention to identify the control points and rate-limiting steps that confer greatest vulnerability. In Aim 2, we will further define the significant vulnerability of the NRel state in OM through changes in release kinetics, gene expression and clearance of otopathogenic biofilms by innate immune effectors. In Aim 3, we will conduct preclinical testing of specifically targeted antibodies capable of inducing the NRel state to resolve experimental OM due to NTHI, the predominant pathogen of acute OM, recurrent OM, chronic OM, CSOM and OM that fails treatment.