Boston Children'S Hospital

NIH Research Projects · FY 2026 · 2026-01

Abstract/Project Summary As an essential component of most behaviors, the sympathetic system regulates many vital physiological processes, such as heart rate, blood pressure, and respiration. Sympathetic activities exhibit an arousal state- dependent basal level (tone), which is subject to behavioral context-dependent regulation (activation and inactivation). In contrast to its importance, much less is known about top-down sympathetic regulation. Our recent studies reveal several populations of spinal projecting neurons (SPNs) with the capacity to regulate sympathetic preganglionic neurons. We hypothesize that medullary and hypothalamic SPNs may form parallel circuits to control state-dependent sympathetic activities with or without accompanying motor engagement. The objective of this proposed study is to test this hypothesis by analyzing the fundamental circuit architectures of sympathetic controlling SPNs, and determining the roles of these SPNs in regulating the basal tone of sympathetic activity, as well as their response to exercise and pain. The outcomes of this study are expected to advance the fundamental knowledge about autonomic regulation and its health impact.

NIH Research Projects · FY 2025 · 2025-12

PROJECT SUMMARY Polyamines (PAs) are positively charged, branched small molecules that are ubiquitous throughout the cell, heavily concentrated within the nucleus, and are frequently found bound to negatively charged nucleic acids and proteins. PAs play regulatory roles within the cell due to their ability to bridge molecules intra- and intermolecularly, leading to the condensation and compaction of DNA, nucleosomes, and chromatin. Despite their small size, PAs participate in genomic spatial and topological organization, playing roles in genome integrity and gene expression. With Parkinson’s disease, polyamine homeostasis is perturbed. In addition, DNA damage levels are elevated and are not efficiently repaired, eventually leading to neurodegeneration and cell death. This study will investigate and determine the molecular mechanism of polyamines on DNA dynamics during DNA repair by homologous recombination (HR). It will utilize in vitro single-molecule and bulk solution assays to assess the extent to which PAs impact the flexibility and dynamics of single-stranded, double-stranded, and non- canonical DNA structures to understand how PAs can impact the dynamics of their formation and accumulation (Aim 1). The in vitro assays will be extended by adding RAD51 to understand the role of PAs in protein-DNA interactions and their impact on RAD51-mediated strand exchange and repair (Aim 2). This will be followed by live cell fluorescence microscopy and chromatin immunoprecipitation sequencing of RAD51-mediated repair in a PA-dependent manner to visualize and quantify the role of PAs in DNA damage repair (Aim 3). The combination of Sua Myong and Taekjip Ha will foster a strong training environment. The Myong lab is one of the leaders in studying non-canonical nucleic acid conformations and protein-nucleic acid interactions leveraging novel single-molecule and cellular approaches. The Ha lab is also a leader in visualizing and quantifying genome maintenance and mechanics by developing and using fluorescence imaging approaches in cellulo combined with CRISPR technology. The mentorship team of both Sua Myong and Taekjip Ha will cover the biophysical and genome topics necessary to build a strong academic career in tackling health questions through a basic science research lens.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT: Respiratory viral infections are associated with significant morbidity in both children and adults. While genomic sequencing of wastewater samples has emerged as promising means of monitoring emerging viral strains, school-based sampling remains largely unexplored. The main objective of this proposal is to validate usage of viral sequencing of dust from elementary schools for early detection of emerging viral strains. The central hypothesis for this study is that viral metagenomic sequencing of dust samples obtained from elementary schools can be used for viral surveillance and identify key viral strains that can affect key clinical outcomes such as school absenteeism. The rationale for this project is that high viral diversity exists in elementary schools, and school-based environmental sampling can be more sensitive than clinical sampling at detecting viral strains. The central hypothesis will be tested through two specific aims: 1) evaluate early detection of emerging viral genomes using viral metagenomic sequencing of classroom dust samples; and 2) identify viral strains that best predict school absenteeism. To achieve these aims, an ancillary study will be designed based on the School Inner City Asthma Intervention Study (SICAS-2: ClinicalTrials.gov NCT02291302) using 571 dust samples longitudinally collected from 209 classrooms across 41 schools from 2015-2019. Dust samples will undergo viral metagenomic sequencing using a hybrid-capture approach. Under Aim 1, phylogenetic and time-based analyses will be used to compare viral genomes from school dust samples to clinical genomes in existing national databases. For Aim 2, predictive models using machine learning will determine the likelihood of increased absenteeism based on the presence of specific viral strains. This project uses an innovative sequencing approach to explore school environmental sampling for viral surveillance, offering critical insights to advance targeted public health interventions and reduce exposures. The long-term goal of the candidate is to develop expertise in virome sequencing in order to study the impact of environmental exposures on pediatric respiratory health. To accomplish this, the training plan will be focused on building skills in bioinformatics, machine learning, advanced biostatistics, and scientific and grant writing. This will be accomplished through formal coursework, collaborative work, conference participation, and mentorship from experts in the field, paving the way for a career development application and ultimately, an independent scientific career.

NIH Research Projects · FY 2025 · 2025-09

Abstract Although autism is known to be highly heritable, the lack of 100% concordance in monozygotic twins and stronger concordance between dizygotic twins over full siblings offers strong evidence for environmental susceptibility factors. There is accumulating evidence for several non-genetic factors, including air pollution, pesticides, heavy metals and pregnancy-related complications. However, studies of environmental susceptibility have been limited by modest sample sizes, indirect measurement and incomplete characterization of exposures, and lack of a multi-dimensional approach that accounts for the interplay between genetic and environmental factors that impact the probability of autism as well as its heterogenous trajectories. The infrastructure already in place in SPARK, a large, US-based cohort of >150,000 recontactable people with autism, offers a unique opportunity to implement a large-scale integrative study design to identify prenatal and early life exposures that impact autism and how exposures interact with genetic risk. Our long-term goal is to identify exposures that influence autism and understand the molecular mechanisms by which these exposures impact the genome, epigenome, and metabolome. The objective of this proposal is to characterize prenatal and early life exposures in >20,000 children with autism and identify exposures that impact probability of autism, developmental trajectories in autism and response to behavioral intervention. We will also investigate the interaction between environmental exposures and genomic risk. In our first aim, we will use multivariate regression and machine learning methods to identify geospatial exposures associated with social communication abilities and response to educational/behavioral intervention in thousands of individuals with autism. In the second aim, we will evaluate gene-environment interactions using the geospatial exposome and the entire distribution of autism genetic risk variants, including common and rare variants, in thousands of individuals with autism in SPARK. In the third aim we will directly measure exogenous exposures and endogenous biological responses in the perinatal period by performing untargeted high-resolution exposomics on residual newborn blood spots from a subset of the SPARK cohort. We will also perform long-read DNA sequencing to assess DNA methylation epigenetic signatures associated with environmental exposures. Ultimately, our goal is to build a model that can help clinicians and families assess genetic and non- genetic probability of autism and predict response to treatment to maximize the potential of individuals with autism. The findings from this study will be significant because this will be the first large-scale investigation that comprehensively evaluates the interplay between exposomic and genomic risk factors in autism and will yield novel insights into the mechanisms that drive risk and resilience in this complex condition. Figure 1: Study overview. Residential history will be collected in 20,000 children with autism and siblings born after 2000. We will investigate environmental interactions with the genome, epigenome and metabolome.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT A-T is a pediatric neurodegenerative condition caused by recessive mutations in ATM, a gene which encodes a critical element of the cellular DNA damage response. Individuals with A-T suffer from gradual loss of muscle coordination due to progressive cerebellar degeneration, leading to loss of ability to swallow, read, communicate, and ambulate. There are currently no known effective treatments for this severely debilitating condition, and it is typically fatal by young adulthood. Here we study the safety and effectiveness of a novel genetic treatment strategy for ataxia telangiectasia (A-T). Antisense oligonucleotides are 15-25 nucleotide snippets of synthetic RNA-like molecules that distribute broadly in the brain and spinal cord when administered via intrathecal injection. They bind to specific genomic targets via Watson-Crick basepairing, and can be designed to change patterns of gene splicing in therapeutically useful ways. 15% of individuals with A-T have mutations that could be treatable using splice-modulating ASOs. Here, we test the clinical therapeutic utility of atipeksen, a 22 nucleotide ASO designed to correct the impact of ATM c.7865C>T, a recurrent mutation that causes disease by creating an abnormal splice site in ATM exon 53. Atipeksen treatment of cell lines from A-T patients bearing this mutation inhibits use of this abnormal splice site, restoring normal ATM splicing and rescuing normal gene function. The current clinical trial is designed to test the safety and effectiveness of atipeksen in A-T in individuals bearing the ATM c.7865C>T mutation. The goal of this study is to slow A-T associated neurodegeneration, delay the progression of neurologic symptoms of A-T, and improve quality of life, using neurologic disability rating scales, brain imaging, and exploratory biomarkers as outcomes. Results from treated patients will be compared against a control cohort of untreated A-T patients with ATM c.7865C>T. Success will provide an empirical foundation for advancing additional precision genetic therapies for A-T and other neurodegenerative conditions.

NIH Research Projects · FY 2025 · 2025-09

Project Summary mRNA-lipid nanoparticle (LNP)-based vaccines are an exciting platform that made advances in combating the pandemic viruses. However, currently licensed mRNA-LNP vaccines approaches are limited in their ability to induce long lived antibody durability, degree of protection varies by age, and levels of cell mediated immunity (CMI) are suboptimal. mRNA vaccine self-adjuvantation is lower in aged individuals, with impaired immune activation upon exposure, inducing insufficient immunity with consequential impaired protection. To try to advance solutions, our published and unpublished work focus on two major novel mRNA vaccine innovations. One is a multi-organ protection (MOP) mRNA sequence homologous to tissue-specific micro-RNAs and enable tissue- specific mRNA translation, constraining expression to the site of injection. Secondly, a biological mRNA-encoded molecular adjuvant precisely guides immunity through inducing the potent and Th1-polarizing analyte, IL-12p70. Based on promising SARS-CoV-2 spike-trimer specific humoral and CMI, in young and aged murine models, and protection from viral challenge in non-human primates (NHPs), the goal of this U01 IICT application is to evaluate a mRNA encoding molecular adjuvant, delivered alongside MOP technology, in the context of a SARS-CoV-2 vaccine. Spike-MOP (CTx892) and IL-12-MOP (CTx672) are first-in-human evaluations, so to assess safety and tolerability, we would first perform dose escalations for CTx892 (stage 1A) and CTx 672 (stage 1B). Based on tolerability, doses less than maximal tolerated CTx892 are chosen to expand sample size for subsequent immunogenicity and mechanism studies. In stage 2, two different doses of CTx 892 will be paired with an escalating CTx672 (IL-12-MOP, e.g., 0.1, 1 µg) dose to assess adjuvanticity along with local and systemic safety and tolerability. Additionally, humoral immunity will be quantified with Spike-specific total IgG (titer and pre-immunization fold- change), IgG1 and IgG3 (Th1 markers), and IgG4 (Th2 marker), with surrogate virus, pseudo- virus, and true viral neutralization. Cellular immunity will include innate assessment in the days after, and adaptive CD4, CD8, and B cell immunity in the weeks, post-immunization. Further readouts include dried blood spots in the hours post-injection to assess proteomic kinetics. In Stage 3, biopsies of lymph node at ~14 days post-immunization and bone marrow at 6 months post-immunization will enable in-depth immunophenotyping and quantification of long-lived plasma cell (LLPC) responses. Durability of humoral and cellular immunity will be probed with a 6-month follow-up sampling of peripheral blood, IL-12-sustained humoral and cellular immunity.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT/SUMMARY: The hereditary spastic paraplegias (HSPs) are a diverse group of over 80 neurodegenerative disorders characterized by progressive spasticity and weakness of the lower extremities, leading to significant disability and reduced quality of life. Despite the growing understanding of the genetic basis of HSPs, therapeutic options remain limited, leaving a critical unmet medical need. The Spastic Paraplegia Centers of Excellence - Research Network (SP-CERN) - RDCRC is a collaborative consortium designed to accelerate the development of effective treatments for HSPs by uniting leading researchers, clinicians, and patient advocacy groups. SP-CERN will focus on three primary objectives: (1) advancing research to identify and validate therapeutic targets, (2) promoting clinical trial readiness by standardizing outcome measures and biomarker development, and (3) fostering collaboration across institutions to share resources and expertise. The SP-CERN - RDCRC features an administrative core, a pilot project core aimed at supporting innovative ideas and addressing emerging themes dynamically, and a career enhancement core poised to support the next generation of clinician-scientist leaders in the field of rare disease research and HSP. The consortium also supports three longitudinal Clinical Research Projects aimed at creating clinical trial readiness through natural history studies, developing a platform approach to gene therapies, advancing diagnostic yield and gene discovery through new genomic tools and machine learning applications, and using digital biomarkers as novel surrogates for disease progression. This integrated approach aims to bridge the gap between basic research and clinical applications, ensuring that scientific discoveries translate into tangible benefits for patients. Through a combination of innovative research strategies, including genetic studies and clinical trials, SP-CERN will address the complexities of HSPs and pave the way for novel therapeutic interventions. The consortium's efforts will be supported by a robust infrastructure that facilitates data sharing, patient engagement, and collaboration among participating institutions. By establishing the SP-CERN RDCRC, this project will create a sustainable framework for the development of therapies that can improve the lives of individuals affected by HSPs, ultimately contributing to a reduction in the burden of these disorders on patients, families, and the healthcare system.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Hematopoiesis involves stem and progenitor cell progression to the mature lineages. Unlike the steady production of new red cells in homeostasis, progenitors must rapidly regenerate many red blood cells under stress conditions. Throughout evolution, the erythroid program has adapted cell-specific processes for function. For example, translation regulation has also been adapted to ensure high levels of globin expression. However, the molecular mechanism facilitating the efficient translation of erythroid mRNAs has not been fully investigated, particularly during stress response. We found that stress granules (SGs) form in erythroid cells in multiple conditions, including in mice during phenylhydrazine-induced hemolytic anemia and in human CD34+ KO of the RNA demethylase, ALKBH5. ALKBH5 KO hCD34+ cells showed defective erythropoiesis measured by in vitro colony formation assays. Proteomic analysis of these cells showed significantly decreased ATXN2 levels. ATXN2 is a stress granule (SG) core component involved in translational regulation and RNA metabolism. ATXN2 overexpression (OE) in wild type erythroid progenitor cells led to accelerated erythropoiesis. STED microscopy of ALKBH5 KO K562 erythroid cells showed massive SGs containing SG structural proteins and m6A-modified RNAs. Restoring ATXN2 levels by lentiviral OE in the ALKBH5 KO cells dissolved SGs and rescued the blocked erythroid differentiation. We investigated the ATXN2 interactome in normal and stress erythroid cells by anti-ATXN2 pull-down mass-spectrometry and high-resolution microscopy. We discovered a novel ATXN2-SRSF RNA binding complex (primarily SRSF8) only in normal erythroid cells. SRSF proteins are involved in RNA metabolism and translation processes. In ALKBH5 mutant cells, ATXN2 bound SG proteins but not SRSF factors. hCD34+ cells with defective SRSF8 showed blocked erythroid differentiation, and the SRSF8 function is required for the ATXN2-OE rescue of erythropoiesis of ALKBH5-KO CD34+ cells. We propose that the ATXN2-SRSF8 complex is essential for the constant delivery of mRNAs to the ribosome in normal erythropoiesis. Under stress, the ATXN2-SRSF8 complex dissociates, and the complex-associated mRNAs are sequestered into SGs along with the ATXN2 protein. During the stress resolution phase, the ATXN2-SRSF8 complex reassociates to load SGs-stored mRNAs onto ribosomes. We will study how ATXN2 and SRSF8 work together at the molecular level to load erythroid-specific RNAs onto the ribosome for efficient translation and accelerate erythropoiesis following stress. We studied the erythropoietic phenotype of ATXN2 mutant mice. ATXN2 mutant mice have SGs, show abnormal red cell size and have defects in PHZ-induced recovery. We will investigate erythropoiesis in mice overexpressing ATXN2. We have made ATXN2 mutant zebrafish to probe SG formation and function in vivo. We plan to evaluate ATXN2 and SGs activity in erythroid cells derived from anemic patient CD34+ cells. Our functional analysis of ATXN2-SRSF8 complex and SGs in erythropoiesis will improve the understanding of normal- and stress-erythropoiesis and may lead to novel therapies for erythroid disorders.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Respiratory diseases often remain inadequately treated due to a limited understanding of the cellular composition and developmental programs within the lung. Recent single-cell studies have identified novel cell types and progenitor cells in the human airway, revealing potential new therapeutic targets. A particularly intriguing discovery is the pulmonary ionocyte, a rare cell type marked by the expression of FOXI1 and ASCL3 and high levels of the cystic fibrosis transmembrane conductance regulator (CFTR), the gene implicated in cystic fibrosis (CF). These cells, which possess long cytoplasmic projections and interact with other airway cell types, play a debated role in airway function across species. While their involvement in airway surface liquid viscosity and ion transport has been observed in mice and ferrets, such roles remain unconfirmed in human in vitro models. This project aims to deepen our understanding of pulmonary ionocytes using human iPSC-derived airway epithelium. Preliminary data show that FOXI1 deletion (KO) in these models alters epithelial composition, increases secretory cells, and decreases ciliated cells, with a concomitant reduction in CFTR-mediated ion transport. Furthermore, FOXI1 overexpression (OE) can reverse these defects, and the introduction of wild-type CFTR ionocytes into F508del homozygous backgrounds restores chloride transport. Additionally, our findings highlight a temporal role for NOTCH signaling in ionocyte development, where combined inhibition of NOTCH and activation of Sonic Hedgehog (SHH) enhances ionocyte numbers. Our research has three primary aims: 1) Elucidate Ionocyte Differentiation Pathways: We will manipulate SHH and NOTCH pathways to increase ionocyte numbers, investigate the mechanisms through single-cell RNA sequencing, and perform chemical screens to identify additional signaling pathways and surface markers for ionocyte enrichment. 2) Investigate Ionocyte Function in Airway Ion/Fluid Transport: We will use FOXI1-KO and FOXI1-OE iPSC lines to assess ionocyte impacts on airway ion/fluid transport and mucociliary clearance. We aim to determine whether FOXI1 overexpression can rescue deficiencies in FOXI1-KO ionocytes and if wild-type ionocytes can restore ion transport in CF airways. 3) Explore Ionocyte Interactions with other airway cell types: Through single-cell RNA sequencing, fluorescent reporters, and high-resolution live-microscopy, we will examine how ionocytes interact with neighboring airway cells and track ionocyte emergence during differentiation. Our project seeks to provide insights into the regulatory mechanisms governing ionocyte differentiation and function, which could pave the way for new therapeutic strategies in treating CF and other airway diseases.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Asthma is the most common chronic disease of childhood in the United States, causes significant morbidity, and accounts for billions of dollars in health care utilization, despite aggressive measures to identify remediable causes. Radon is the most important natural source of human exposure to ionizing radiation. Rn toxicity is due to its decay products that emit alpha (α), beta (β), and gamma (γ) radiation that form ultrafine clusters that attach to airborne particles rendering particulate matter (PM) radioactive. Radon sourced primarily from the adherence of airborne radioactivity to particulate matter, is common in US homes due to the broad geologic presence of radon in the earth's crust. Inhaled radon it associated radioactive PM deposits in the airways and alveolar regions damaging nearby cells. Recent evidence from our lab and others has identified radon as a key contributor to asthma and chronic obstructive pulmonary disease morbidity and mortality beyond the known carcinogen effects. These adverse health effects are seen at levels below the current World Health Organization recommended exposure thresholds for radon mitigation. We have collaborated for over 15 years to investigate environmental hazards in schools and homes of children with chronic respiratory diseases. From these studies, we have generated preliminary data that demonstrate: i) indoor Rn is associated with asthma symptoms and airway inflammation in children and ii) it is well-established that radon mitigation systems significantly reduce radon by 90% and thereby reduce particle radioactivity. Although our data is promising, the impact of radon mitigation on improving health outcomes can only be answered in a clinical trial. The results of our study could impact clinical practice. Our overall hypothesis is that reduction of radon in homes will result in significant reduction of asthma symptoms and inflammation in children with asthma. We propose a 48 week randomized, sham controlled, radon mitigation intervention trial in 180 children with asthma who live in high and low radon geologic exposure areas to 1) determine the effect of radon mitigation in the primary living space on asthma symptoms, primarily, as well as lung function; and 2) determine whether radon mitigation will reduce established biomarkers of inflammation, specifically airway FENO and established inflammatory biomarkers (IL4,5,13, 6, and C-Reactive Protein (CRP)). This study is an unprecedented, high impact opportunity to test a personalized remediation strategy for a ubiquitous but novel harmful respiratory exposure. Determining the benefit of radon interventions offers a precise and personalized mitigation strategy to improve asthma morbidity. These findings will inform public health policy regarding the risks of residential radon exposure in children with chronic health conditions and thresholds for mitigation. It will allow us to identify health burdens resulting from real-life indoor exposures and associated health effects among vulnerable children with asthma. If successful, this will provide guidance on cost-effective, environmentally-friendly, and scalable technologies to improve indoor air quality to reduce disease burden in an area of significant unmet need in children with chronic airway inflammation and asthma.

NIH Research Projects · FY 2025 · 2025-09

Modified Project Summary/Abstract Section Most sexual minority adolescents and young adults (AYA) are attracted to more than one sex or to people regardless of sex (e.g., bisexual). Limited prior research suggests that bisexual AYA are at greater risk than their lesbian, gay, and heterosexual peers for adverse mental and behavioral health (MBH) outcomes, in part due to anti-bisexual stigma, a unique form of stigma enacted by both heterosexual and lesbian/gay individuals and communities. We will use a community-engaged sequential mixed methods study design to: (1) Examine how sexual orientation-based stigma is experienced by bisexual AYA. We will collect data via asynchronous online focus groups from AYA who are attracted to more than one sex. (2) Investigate the longitudinal impact of sexual orientation-based stigma on MBH outcomes considering developmental and sociodemographic factors. We will collect longitudinal survey data from bisexual AYA and will recruit a parent/caregiver among a subsample of youth to examine family-related factors that impact bisexual AYA MBH. (3) Determine key protective factors that reduce the adverse impact of sexual orientation-based stigma on MBH outcomes over time. We will utilize Aim 1 focus group data and Aim 2 survey data to examine the role of protective factors in mitigating the adverse impacts of stigma on MBH outcomes among bisexual AYA. The expected outcomes will be a novel understanding of how sexual orientation-based stigma is experienced by AYA, its longitudinal impact on AYA’s MBH, and how protective factors may prevent adverse MBH outcomes in this population. This research is a critical step toward preventing health disparities and improving the health and transition into adulthood for vulnerable AYA.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY The rising incidence of type 2 diabetes (T2D) highlights a growing need to understand mechanisms behind highly-effective therapies such as Roux-en-Y gastric bypass surgery (RYGB) in order to develop better, more widely-applicable treatments. Adaptation in the jejunum (Roux limb) is a hypothesized contributor to diabetes improvement after RYGB, and this is supported by our lab’s work demonstrating a relationship of jejunal adaptation with both glycemic control and T2D remission in humans. Our long-term goal is to understand how changes in GI anatomy effect the powerful, durable shift in energy homeostasis observed after bariatric surgery. The overall objective of this application is to understand how changes in GI anatomy elicit the powerful, durable shift in energy homeostasis observed after bariatric surgery. Our preliminary data demonstrate segment-specific activation of LXR signaling in Roux limb of mice and humans after RYGB which is lost in humans without T2D remission, as well as downregulation of PPARa signaling and enrichment of lipids, especially plasma membrane lipids, in Roux limb. The central hypothesis is that that alteration in intestinal epithelial lipid homeostasis, driven by LXR and PPARa signaling, is necessary for RL adaptation and associated improvements in glucose homeostasis after RYGB. The rationale for this project is that de novo lipogenesis could be a pathway for local intestinal glucose utilization, which is supported by our preliminary data. These experiments will allow us to directly test the role for lipogenic pathways to contribute to intestinal adaptation and metabolic homeostasis more broadly. The central hypothesis will be tested in two specific aims: (1) to test the hypothesis that LXR- and PPARa- mediated lipogenesis drives RL adaptation and glucose regulation in diet-induced obesity; and (2) To test the hypothesis that changes in lipogenesis drive intestinal epithelial adaptation in a disease-relevant in vitro model system. We will use mouse models including our murine model of bariatric surgery in Aim 1; Aim 2 will use a jejunal organoid model derived from human patients with obesity, who are undergoing bariatric surgery. This project will shed light on the key intestinal epithelial cells responsible for Roux limb adaptation and ultimately, potential drug targets that could mimic the beneficial effects of RYGB.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Hematopoietic stem and progenitor cells (HSPCs) closely interact with the niche microenvironment, allowing a dynamic regulation on proliferation, differentiation and self-renewal. Various niche cells have been identified, yet the secreted signaling factors in the hematopoietic niche are less well-defined, limiting the application of HSPC for ex vivo expansion and enhanced engraftment protocol for blood disease treatment. To identify novel niche factors that stimulate HSPC production, I performed an in vivo small molecule chemical screen and identified an antagonist of cysteinyl leukotriene receptor 1 (CysLTR1) that stimulates HSPC divisions that specifically occurs within a ‘pocket-like’ structure that is formed by a group of endothelial cells (ECs) in the niche. The HSPC is known to physically interact with these ‘pocket’ forming ECs and the event is termed ‘endothelial cuddling’. A HSPC division is often observed after endothelial cuddling, yet the mechanism is unknown. I performed single cell RNA-sequencing (scRNA-seq) analysis of zebrafish hematopoietic tissues and identified that CysLTR1 is specifically expressed in a subset of ECs with high expression of niche EC genes. I generated a stable cysltr1:mCherry reporter fish and found that cysltr1:mCherry+ ECs preferentially interact with HSPCs. I generated a stable cysltr1 knockout fish and it phenocopied the antagonist perturbations. Cysteinyl leukotrienes (CysLTs) are lipoxygenase (LOX)-derived lipid mediators. I further performed lipidomic analysis of zebrafish hematopoietic tissues and identified a series of LOX-derived lipid mediators. Interestingly, I performed scRNA- seq analysis and found that corresponding receptors of these lipid mediators were expressed in distinct cell types such as macrophages, neutrophils or stromal cells, indicating unique mechanisms. LOX-derived lipid mediators play key roles in inflammation. However, the role of LOX-derived lipid mediators in the hematopoietic niche remains unexplored. Given my preliminary data, I hypothesize that LOX-derived lipid mediators remodel hematopoietic niches by altering cellular interactions and secreted cytokines to regulate hematopoiesis. Under the supervision of Dr. Leonard Zon, the field-leading expert in hematopoiesis and zebrafish genetics, I will test my hypothesis using high resolution live cell imaging, chemical biology and CRISPR/Cas9 mutagenesis. I have put together a panel of scientific advisory committees who are field-leading experts in vascular biology, niche cell biology, and lipid mediator biology. Together with Dr. Zon, I will receive constant feedback on my research and career development. This K99/R00 award will enable me to gain trainings and acquire skills in scientific training and career development that prepare me to start my laboratory. My overarching goal is to investigate molecular mechanisms of niche-mediated regulations in hematopoiesis in my own laboratory. I aim to contribute my research discoveries to develop enhanced HSPC ex vivo expansion protocol through recapitulating the functional niche signaling and to improve the efficiency and safety of therapeutics through targeting/priming the hematopoietic niche.

NIH Research Projects · FY 2025 · 2025-09

Although injuries are the leading cause of death and disability among U.S. children, timely and accurate injury surveillance does not exist. The long-term goal is to decrease pediatric injuries by analyzing trends and changes over time of important injury mechanisms. The overall objectives in this application are to create a novel pediatric injury surveillance data network using natural language processing (NLP) in a multi-site emergency department (ED) electronic health record (EHR) data system, the Pediatric Emergency Care Applied Research Network (PECARN) Registry. The central hypothesis is that a timely and accurate pediatric injury surveillance system can be created by applying an NLP graphical user interface (GUI) in ED EHR data as soon as it is available (2 months after ED visit). The rationale for this project is that given current data gaps, creating a multi-site ED based injury surveillance system, as most critical trauma and critical illness are treated in EDs, is needed for timely identification of injury trends. The central hypothesis will be tested by pursuing three specific aims: 1) Establish a near-real time pediatric ED injury surveillance system, which is more accurate and timely than current sources, using NLP in multi-site ED data; 2) Determine near real-time trends of pediatric injuries using the PECARN Registry; and 3) Demonstrate feasibility of local pediatric injury surveillance with on-site application of NLP methodology to internal EHR data at 1 hospital, not using PECARN Registry data. Under the first aim we will apply an NLP GUI to the PECARN Registry to identify 3 specific injury mechanisms (suicide attempts, opioid overdose injuries, micromobility device/scooter injuries). We will compare test characteristics for identifying these injuries with NLP to only using diagnosis codes. For the second aim we will report trends and changes over time for these 3 injury mechanisms. For the third aim we will apply the NLP GUI only to internal EHR data at 1 hospital for local injury surveillance of the same 3 mechanisms with more timely data (1 week after ED visit). The research proposed in this application is innovative, in the applicant’s opinion, because it uses physician domain expertise on injuries applied to NLP methodology, which is brought to the extant data sources. The proposed research is significant because it is expected to create a timely, accurate pediatric injury surveillance system to advance prevention of critical trauma and critical illness from injuries.

NIH Research Projects · FY 2025 · 2025-09

How is growth and proportion regulated during development to ensure appropriate scaling, or proportion of elements within a structure? Our lab used genetic screens in zebrafish to identify the potassium channels KCNH2 and KCNK5 as sufficient to cause overgrowth of appendage structures in a coordinated fashion. Further, we investigate a unique presentation of growth dysregulation, macrodactyly, as a case study to understand how integrated, non-cancerous, growth occurs even in the case of potent oncogenic driver mutations. In a broad cohort of patients with macrodactyly, we identified genetic factors associating with patterned digital overgrowth in the PI3K signaling cascade. In parallel, using genetic screens in zebrafish, we identified the potassium channels KCNH2 and KCNK5 as sufficient to cause overgrowth of appendage structures in a coordinated fashion. Of note, we demonstrate functional synergy between PI3K signaling and altered KCNH2 variants identified in patients, sufficient to lead to patterned overgrowth of appendages. These findings highlight unknown role of potassium channels in mediating growth potential of oncogenic mutations. The lack of knowledge on the role of these genetic factors during development limits our current ability to effectively address and alleviate disease progression. To detail the intricate regulation of somatic clone growth, we outline three specific aims that leverage clinical and experimental analyses to address changes in gene regulation, role of potassium channel function in modifying growth potential, and tissue and genetic constraints on clonal dynamics. In Aim 1, we take advantage of gain- and loss-of-function lines to address function of the potassium channels Knch2 and Kcnk5 in regulating differentiation, growth and proportion of the limbs. These mouse models permit detailed tissue-specific analysis of potassium channel function mirroring mutant analysis in the zebrafish. Through histological and anatomical analyses, we will assess the effect of altering potassium channel function on the differentiation of osteochondral progenitors in long bones of the developing limb. In Aim 2, we will use zebrafish models to observe cell behavior in real time to specifically address the role of defined genetic context and developmental constraint in clonal expansion during development. We will extend and complement this work in Aim 3 to systematically identify genetic regulators of growth using new unbiased, zebrafish somatic-mosaic screens to identify modifiers of overgrowth regulation. Through these integrated, and separate innovative approaches, we can distill the mechanisms by which overgrowth is shaped, and potentially alleviated.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT Limited therapies are available for the preservation of long-term cardiac function following myocardial infarction (MI), a leading cause of death worldwide. The long-term goal is to develop an independent research career as a cardiac surgeon-scientist focused on accelerating the design of novel cell-based therapies for the treatment of MI and ischemia-reperfusion injury (IRI), minimizing infarction-related morbidity and mortality. The overall objective of this proposal is to determine the in-vivo efficacy of myocardial injected human-induced pluripotent stem cell-derived cardiomyocytes and vascular endothelial cells (hiPSC-CMs/vECs) engrafted in retention scaffolds in a post-MI rat model. The central hypothesis is that hiPSC-CMs and hiPSC-vECs in gelatin methacryloyl hydrogel (GelMA-H) engrafted into infarcted cardiac tissue will enhance cell retention, optimizing reestablishment of myocardium and perfused vasculature, leading to reduced fibrosis and preserved cardiac function. The rationale for this research is that rigorous preclinical evidence supporting the therapeutic efficacy of engrafted hiPSC-derived cells would offer a strong scientific framework to guide new strategies to address IRI, enhancing length and quality of life post-infarction. The central hypothesis will be tested via the research aim: Determine the (a) tissue and (b) organ therapeutic efficacy of hiPSC-CMs and hiPSC-vECs engrafted in GelMA-H in an immunodeficient rat model of MI. Under this aim, hiPSC-derived cells will be engrafted in GelMA-H scaffolds and myocardially delivered post-MI. Cell retention will be quantified via bioluminescence. After 4-week survival, immunohistochemical techniques will be employed to elucidate extent of ventricular fibrosis, remuscularization, and angiogenesis and arrhythmogenicity and cardiac function will be assessed. This aim will be accomplished via training goals: 1) Develop proficiency in design and implementation of cell- based translational models and acquire enhanced knowledge in cell differentiation and immunohistochemical techniques; and 2) Attain advanced skills in data collection, analytic strategies, research dissemination, and grantsmanship. Courses, seminars, and conferences will be attended, each purposely selected to accomplish the overall objective and advance the applicant towards an independent basic/translational cardiovascular research career. The research proposed is innovative because it incorporates the use of two cell types derived from hiPSCs engrafted in a hydrogel retention scaffold to maximize the therapeutic effect of stem cell therapy based on the demonstrated benefits of each of these individual components. The proposed work is significant because it is expected to: 1) Provide a strong potential to guide new strategies to address IRI; 2) Mitigate healthcare disparity and burden of a pervasive, preventable public health issue; and 3) Provide necessary training required to submit a competitive K08 focused on development of preclinical interventions to treat MI or IRI as an important step toward future use in humans. Ultimately, such knowledge will have positive impacts on the development of novel cell-based therapies that improve outcomes of patients experiencing MI or IRI.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Calcium ion (Ca2+) play a crucial role in regulating striated muscle contraction. Intricate nanoscopic structures, known as dyads in cardiomyocytes, efficiently coupling membrane excitation with Ca2+ release and sarcomere contraction. However, the mechanisms that regulate the activity of dyads remain poorly understood. Ryanodine receptor (RYR2) is the dyadic component responsible for the majority of intracellular Ca2+ release in cardiomyocytes. Dysregulation of RYR2 contributes to the pathogenesis of several human diseases, including heart failure and arrhythmias. For example, dominant variants in RYR2 cause catecholaminergic polymorphic ventricular tachycardia (CPVT), in which excessive diastolic Ca2+ release triggers potentially lethal cardiac arrhythmias. We discovered that CMYA5 (Cardiomyopathy-associated protein 5) is a little studied component of dyads. CMYA5 ablation disrupts dyad organization and positioning, resulting in cardiomyopathy and abnormal Ca2+ release. In our preliminary data, we show that CMYA5 interacts with RYR2 via an RYR2 interaction domain (RID). RID binding significantly reduces the open probability of both wild-type and CPVT mutant RYR2 channels. Based on these findings, we hypothesize that CMYA5 is a novel endogenous regulator of RYR2 channel activity. Targeting RID-mediated inhibition of these channels could potentially treat CPVT caused by RYR mutations. To test this hypothesis, we will investigate CMYA5-RID’s role in regulating RYR2 activity in cardiomyocytes (Aim 1), and test its therapeutic potential to reduce aberrant Ca2+ release and arrhythmias in CPVT (Aim 2). This study is significant as it will elucidate the physiological roles of CMYA5 in modulating ryanodine receptor activity, and explore therapeutic strategies based on CMYA5 inhibition of RYR2 activity. These efforts will lay the groundwork for development of more effective therapies for more common forms of heart disease.

NIH Research Projects · FY 2026 · 2025-09

Iron plays a vital role in brain development, contributing to processes such as synaptogenesis, myelination, and neurotransmitter synthesis. Perinatal iron deficiency has been linked to delayed nerve conduction, impaired recognition memory, motor difficulties, and lower overall developmental scores. Preterm births affect about 1 in 10 U.S. infants, and 25 - 85% of them show signs of iron deficiency during infancy. Accurate measurement and timely supplementation of brain iron during this early critical phase is therefore essential for optimizing neurodevelopmental outcomes. Qutantitatve susceptibility (QSM) and R2* MR imaging have proven to be important tools for the non-invasive quantitative measurement of brain iron levels. However, these techniques have not yet been robustly deployed in infant studies, especially those preterm infants. The key challenges include: 1) Traditional Cartesian imaging techniques are lengthy (>10 mins) for neonates and highly motion-sensitive; 2) Motion-induced geometric and field distortions introduce additional phase errors, compromising quantitative accuracy; 3) Standard QSM captures average susceptibility within each voxel, making it difficult to separate iron (positive susceptibility) from myelin (negative susceptibility) in the neonatal brains. To address these challenges and enable the reliable use of quantitative MRI techniques to study iron deficiency of neonatal brains, we propose motion-robust 3D kooshball data acquisition and advanced reconstruction strategies for rapid high-resolution QSM and R2* imaging of neonatal brains. In Aim 1, we will extend an existing 3D radial sequence to incorporate multi-echo readouts, adapting previously developed motion estimation algorithms that extract brain motion and motion-induced field maps from the data itself. We will further design an image space-based reconstruction algorithm to generate high-quality MR images from undersampled radial data. In Aim 2, we will develop and validate a motion-corrected, deep-learning- regularized model-based nonlinear reconstruction algorithm that directly estimates quantitative maps from highly undersampled radial data using our rapid protocol from Aim 1, significantly reducing the imaging time. In Aim 3, we will apply source separation techniques to distinguish iron (positive) and myelin (negative) susceptibility from the estimated QSM maps using R2* and R2 information. We will perform evaluation of the utility of the developed technologies in assessing brain iron development in preterm neonatal subjects with iron deficiency compared to full-term neonates. To that end, our novel technologies would speed up the quantitative MR exams and allow for accurate and reliable iron quantification of newborn brains.

NIH Research Projects · FY 2025 · 2025-09

Project Summary: Pediatric in-hospital cardiac arrest (p-IHCA) is a devastating medical condition associated with high mortality and morbidity. While the American Heart Association provides recommendations to guide resuscitation efforts, most of these are derived from low-quality observational evidence with critical risk for bias. This is because randomized controlled trials (RCT) for intra-arrest interventions are exceedingly difficult to perform. Given the critical need for high-quality RCTs to improve outcomes in this field, new trial design techniques must be developed to help overcome these barriers. One key element of trial design that may be modified to enhance feasibility is the selection of the primary outcome. Traditional endpoints for cardiac arrest studies include return of spontaneous circulation (ROSC), survival to discharge and survival to discharge with favorable neurologic outcome (FNO). However, these outcomes are all problematic for two reasons. First, the binary nature of these endpoints leads to the need for very large sample sizes to detect differences in outcomes. This is especially cumbersome in p-IHCA because not only is it a rare event, but also because the minimal clinically important difference for clinicians treating cardiac arrest is very small, owing to its high lethality. Second, these outcomes are limited in their ability to reflect the success or failure of resuscitation efforts. For example, ROSC reflects a successful circulatory resuscitation, but there is no indication of whether the patient survives to hospital discharge, with or without FNO. Survival and survival with FNO represent more patient-centered outcomes, but they often do not reflect the results of the resuscitation in isolation given the vast number of life-limiting disease states associated with p-IHCA as well as the risk for unrelated complications in the interval between arrest and discharge. Therefore, we propose to explore the use of Time to ROSC as a novel outcome for pediatric cardiac arrest studies. Time to ROSC is a critical element in resuscitation as it represents the period of the “low flow” circulatory state to critical organs including the heart and brain and is strongly associated with survival and survival with FNO. To develop this outcome, we plan to use a large national pediatric cardiac arrest registry to perform simulation-based analyses to determine the optimal statistical strategy for patients who do not achieve ROSC. Following the development phase, we will use data from a recent multicenter trial to evaluate the performance of Time to ROSC under “real world” conditions. As power is likely to increase with this time-to- event analysis, this approach has the potential to significantly decrease the number of patients needed to detect meaningful differences between treatment groups as well as better reflect the direct effects of intra- arrest interventions, making intra-arrest trials more feasible, informative and cost-effective.

NIH Research Projects · FY 2025 · 2025-09

This proposal is for the transformation of a human-induced pluripotent stem cell (iPSC)-derived bone marrow organoid for the study of hematopoiesis. The scientific premise of this project is to help overcome the limitations of current hematopoiesis research models, such as two-dimensional cell cultures and animal models, by utilizing a human bone marrow organoid. Indeed, organoids have the potential to mimic the microenvironment and functions of tissues or organs more accurately, bridging the gap between in vitro and in vivo systems. We have created a bone marrow organoid using iPSCs that replicates key marrow elements like sinusoidal vessels, stroma, megakaryocyte/endothelial interactions, and myeloid cells. This organoid has already shown promise in studying hematologic cancers and therapeutic interventions. However, the potential of the bone marrow organoid extends beyond malignant samples. In the proposed project, we aim to revolutionize the organoid for the study of hematopoiesis in three specific aims. The first aim is to perform cellular mapping of the bone marrow organoid over its development using advanced flow cytometry and microscopy techniques. This will provide valuable information about the changes that occur in the organoid over time and serve as a resource for the wider hematopoiesis field. The second aim is to optimize the bone marrow organoid for the study of hematopoietic development. The current protocol, which focuses on the differentiation of hematopoietic stem and progenitor cells (HSPCs), leads to a progressive loss of HSPCs over time. By incorporating knowledge from animal models and cell culture systems, we will develop an iPSC differentiation protocol that maintains a stable HSPC population over a 60- day period. This optimization is crucial for the organoid model to effectively study hematopoiesis. The third aim is to culture the organoid under flow using a microfluidics platform. The lack of interstitial flow in the current model limits its ability to replicate physiological conditions. By culturing the organoid in a 3-channel microfluidic bioreactor, we will introduce interstitial flow and enhance vascular maturation and maintenance. This setup will also allow the efficient delivery of small molecules, proteins, and cells into the organoid, mimicking in vivo blood flow. Further, it will present opportunities for testing therapeutic strategies and studying physiological mechanisms relevant to hematopoietic diseases. In sum, this project aligns with the goals of the Catalytic Tool and Technology Development FOA and aims to transform the bone marrow organoid into a novel biological tool that advances translational hematopoiesis research.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Macrophages exhibit remarkable functional heterogeneity and plasticity, reflecting their diverse developmental origins from yolk sac, fetal liver progenitors, or bone marrow. This diversity underpins their roles in tissue homeostasis and inflammation, with tissue-resident macrophages, primarily derived from yolk sac and fetal liver progenitors, involved in homeostatic tasks, and monocyte-derived macrophages from bone marrow driving acute inflammatory responses. Dysfunctions in these macrophage subtypes are linked to varying disease phenotypes, such as atopic dermatitis, graft vs host disease, arthritis, atherosclerosis, and Langerhans cell histiocytosis. The proposed research seeks to delineate the influence of ontogeny on macrophage specification and exploit this understanding to target dysregulated subsets in diseases like atopic dermatitis, Langerhans cell histiocytosis, and graft-vs-host disease. Building on preliminary findings that suggest distinct properties and developmental pathways for macrophages from fetal or adult hematopoietic stem and progenitor cells (HSPCs), and the identification of AHR as a key gene regulating TRM self-renewal, the study aims to selectively target and modulate these macrophage subsets. The approach encompasses culture-based methods, xenotransplantation, and genomic techniques, striving to unveil novel insights and therapeutic strategies for conditions driven by macrophage dysregulation. Dr. Katie Frenis is a 2nd year postdoctoral fellow who aims for a faculty position in the next 2 years. The work above is the cornerstone of the laboratory she hopes to build. Dr. Frenis is interested in the innate immune system, its heterogeneity, how the heterogeneity arises, and how it impacts disease across the age spectrum. She has a strong mentorship and collaborative network and aims to learn lab and budget management, grantsmanship, and administrative details before the R00 phase from her close mentors. Because of her career in teaching and publishing, she has extensive leadership and teaching/training experience and aims to leverage that into becoming a dedicated mentor. She will carry out her postdoctoral phase at the hematology/oncology department at Boston Children’s Hospital, which is a rich environment with a good atmosphere and a proven track record of producing faculty appointees.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Disorders of growth plate chondrocyte maturation impact the growth of the skeleton, resulting in a spectrum of diseases from skeletal dysplasia to extreme short stature. These diverse conditions underscore the importance of tight regulation to normal epiphysial physiology, yet the genetic pathways directing chondrocyte maturation are poorly understood. The current proposal leverages an in vitro model of the growth plate to (1) conduct high- throughput, genome-wide functional KO screening of chondrocyte maturation, (2) prioritize screening hits with orthologues linked to human skeletal growth through genome-wide association studies (GWAS), and (3) investigate the mechanisms by which top screening targets act to affect chondrocyte maturation. During the course of my K08 award thus far, I have conducted a screening assay in which a lentiviral library of 80,000 unique single-guide RNAs (sgRNAs) is transduced into Cas9+ chondrocytes (300 million cells from the MLB13 cell line) to simultaneously KO 20,000 genes in replicate. After four days of maturation, a time at which 95% of cells are usually immature, KO chondrocytes are FACS sorted for the maturation marker CD-200; KOs driving early maturation (CD-200 high) are compared to immature internal controls (CD-200 low). This screening assay can robustly detect genetic determinants of chondrocyte maturation and has already identified genes highly relevant to skeletal biology, including members of the Indian hedgehog signaling family. Interestingly, I found that loss of Protein Inhibitor of Activated STAT1 (PIAS1) results in significantly premature chondrocyte maturation. Not yet known to function in the growth plate, PIAS1 is a highly plausible candidate as it has been shown to modulate the activity of transcription factors in other cell types that also direct growth plate elongation and maturation. In the present application, I intend to investigate mechanisms by which PIAS1 acts to delay growth plate chondrocyte maturation in vivo (Aim 1) and probe a role for PIAS1 in murine limb development (Aim 2). Securing this award will be instrumental in providing crucial support for generating preliminary data essential for a subsequent R01-level grant. This subsequent grant will enable me to conduct an in-depth investigation into the mechanistic role of PIAS1, positioning it as a promising and innovative target in both limb development and growth plate maturation.

NIH Research Projects · FY 2025 · 2025-08

Overall Project Abstract The overall focus of the “Immunization against Multidrug-resistant Pathogens: Activating T Cell Immunity” Center of Excellence for Translational Research (IMPACT-CETR) is to advance promising multicomponent vaccines for Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoniae. These are among the most important bacterial pathogens that cause severe clinical disease and death and yet are becoming increasingly resistant to the most effective antibiotics. The overarching goal of this IMPACT-CETR is to harness the collaborative team’s complementary expertise in immunology, bacteriology, bioinformatics, primatology, vaccine development, and antigen-adjuvant formulations to achieve three deliverables: 1) novel multicomponent vaccines optimized for bacterial proteins and/or polysaccharides that elicit broad and potent serotype- independent protection against S. aureus, P. aeruginosa, and K. pneumoniae infections, including with resistant and MDR clinical isolates; 2) defined key mechanisms of host protection and biomarkers of vaccine efficacy; and 3) nonhuman primate (NHP) models to characterize immunogenicity and surrogate markers of protection. The proposed vaccine components are well-characterized, some are chemically defined, and all are designed for feasible scale-up and manufacture. The three research projects in this CETR are bonded by the theme that tissue-resident memory T cell responses, particularly tissue-resident Th17 cells, are critical for protective vaccines against these pathogens. The translational research projects will develop countermeasures to prevent/reduce disease caused by key resistant and MDR bacterial pathogens. Project 1 addresses a multicomponent S. aureus vaccine formulated with conserved protein antigens using the new and innovative Multiple Antigen Presenting System (MAPS) platform wherein fusion proteins of the avidin derivative rhizavidin with conserved S. aureus protein antigens are complexed to a biotinylated polysaccharide to generate protective B- and T-cells. Project 2 addresses a multicomponent P. aeruginosa MAPS vaccine based on the serotype- independent P. aeruginosa biofilm polysaccharide Psl and its critical lipid epitope, combined with conserved protein antigens including the Th17-eliciting antigen PopB. Project 3 addresses a quadrivalent K. pneumoniae vaccine based on conserved proteins that elicit protective Th17 cells and antibodies. Vaccine candidates will be tested in wild-type and transgenic and/or knock-out mice and in non-human primates. These projects will be supported by an administrative core and three scientific cores that focus on bioinformatics, transgenic mouse models for mechanistic studies, and NHP studies. The expected milestone for each project is the development of a preclinical data package that would pave the way for subsequent IND applications and clinical trials.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The primary vision of this project has been the establishment of investigator and network collaborations that will advance study of the role of the exposome in childhood-onset rheumatic diseases and other autoimmune conditions, with an overarching goal of creating new research resources and forming new teams of pediatric investigators from existing networks that can contribute to the nascent “EXposome in Autoimmune Disease Collaborating Teams” (EXACT) network. We address this focus via collaboration between the EXACT-PLAN consortium (initiated August 2023) and our two existing pediatric networks, which have pre-existing, extensive phenotype and biobank resources: the investigator-sponsored Childhood Arthritis and Rheumatology Research Alliance (CARRA), which sponsors the CARRA Registry, and the NIH/NCATS funded Genomic Information Commons (GIC). Over the first two years of EXACT-PLAN, we have established collaborations that take the initial steps to advance our understanding of the role of exposures in the development and course of autoimmune diseases (AIDs), resulting in a shared definition of `Pillars of Exposome Studies in Autoimmunity' to help drive further EXACT collaborations. At the same time, we have made substantial progress towards fulfilling our aims for the parent project, which are two-fold: (1) Establish a collaborative framework between investigators of the CARRA and GIC networks for augmenting current research activities with exposome data and biobank specimens; and (2) pilot the linkage of publicly available exposome data sets, including air and water quality databases, to existing data sets that the CARRA Registry and GIC maintain. We are further developing and adapting policies, protocols, informed consents, case report forms, and patient surveys that are specifically targeted to incorporate exposome data for childhood-onset rheumatic diseases and other autoimmune conditions. For this supplement, we will further enhance collaborations between our two pediatric networks (CARRA and GIC) with other EXACT-PLAN projects, focusing on the development of geographically diverse pediatric cohorts for exposome-wide association studies, including for both disease-affected and as yet unaffected children and adolescents. To facilitate collaborations with our EXACT-PLAN colleague projects, we follow a near-term, shared vision of creating capabilities to conduct large-scale, exposure-wide association studies and a longer-term focus on achieving the four pillars of exposome studies in autoimmunity that were defined in the initial phase of EXACT-PLAN. For this supplement, as we further collaborate within NIH's planned “EXposome in Autoimmune Disease Collaborating Teams” (EXACT) network, we simultaneously pilot enhanced, longitudinal geographic histories and exposome-ready cohort definition capabilities for the GIC research population at Boston Children's Hospital (BCH). The GIC network is a cooperative, phenotype-genotype biobanking effort with 8 participating pediatric academic medical centers across the US, with broad representation of subjects with differing conditions (14 million subjects, ~24,000 with genome-linked phenotypic data, 180,000 biospecimens collected) and provides search across sites to identify cohorts of individuals with and without rheumatic or autoimmune diseases, with infrastructure also supporting subject recruitment and biobanking for future studies. At BCH, GIC enrolls 2.9 million subjects of which ~7,000 have genome-linked phenotypic data, with ~60,000 biospecimens collected. To enable longitudinal geospatial analyses, we will use the supplement period to explore local administrative linkages for geocoding as well as pilot BCH subject surveys, in a manner that will be readily extensible to other GIC sites outside of BCH. During this project period, we also plan to further engage with other EXACT-PLAN awardees in various capacities, specifically: (1) Continue to collaborate closely on EXACT-PLAN geospatial analyses (John Pearce, PI, Medical University of South Carolina) and establish new collaborations led by this group with the NIH- funded “Network for Exposomics in the United States” (NEXUS; U24ES036819); (2) Identify unaffected pediatric cohorts at the BCH GIC for collaborations with EXACT-PLAN colleagues studying the effect of exposures in pre-clinical rheumatic disease populations (systemic lupus erythematosus, rheumatoid arthritis; Jill Norris, PI, Colorado School of Public Health); (3) Identify and assist with recruitment of BCH GIC and CARRA subjects with Juvenile Idiopathic Arthritis for a remote `ImmuneTracker' mobile app pilot (Wilson Liao, PI, University of California, San Francisco); and (4) Identify pediatric AID cohorts at BCH GIC pertaining to T1DM and celiac disease (Brigitte Norris, PI, University of Colorado). Our multi-disciplinary project team includes researchers in pediatric rheumatology and the exposome, technology experts, and patient representatives. Completion of the aims of this project will further enable new, team-based collaborations to conduct future high-quality, best-practices exposome research for many pediatric autoimmune diseases as part of the future EXACT network.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT: In addition to peripheral symptoms such as skin and kidney disease, many Lupus patients (60-80%) also develop psychiatric symptoms referred to as neuropsychiatric lupus (NPSLE). Recently, we reported an interferon stimulated gene (ISG) signature in the CNS in a strain of lupus mice (Sle1 yaa) that correlated with a behavior phenotype. Using a novel spacial transcriptomic approach (MERFISH), we found discrete patches of cells in the brain and spinal cord that express elevated levels of ISG. The novel finding not only demonstrates IFN- I triggering of neuroinflammation in the CNS but suggests cell activation by IFN-1 is not random but spacially regulated. The discrete patches of cells include clusters of glia, endothelium and neurons expressing elevated levels of ISG. Single nuc RNA seq of the hippocampus and hindbrain, regions important in the behavior phenotype of Sle1 mice, confirmed an overall interferon signature with subclusters of cells expressing elevated levels of ISG including death pathway related genes (Aw et al BBI 2023). To explain the behavior change in the lupus mice, we postulate that IFN-I is secreted locally by border macrophages and infiltrating macrophages recruited by deposits of autoantibodies and activated complement in the small vessels of the brain. In support of neurotoxicity at the patches, preliminary results from analysis of the merfish and single nuc seq data identifies reduced neuronal cell density relative to adjoining non-patch regions. This proposal will examine the patches for synaptic and neuronal cell loss and test the importance of IFN -I activated microglia and astrocytes in mediating the injury. Moreover, it will identify the source of IFN I and how it enters the CNS. The major gaps in knowledge addressed in this study include: 1) How do microglial and astrocyte gene expression and function change in lupus? 2) Is there a link between microglia and astrocyte dysfunction, neuropsychiatric lupus symptoms, and neuronal cell death and/or synapse loss? 3) Is type I IFN a key factor in promoting CNS dysfunction in lupus and what is the source? Two aims are proposed: Aim 1: Test hypothesis that activation of brain glial cells by type I interferon leads to inappropriate synaptic pruning and behavior change. Aim 2. Identify the source of type I interferon in the CNS of lupus mice.