Children'S Hosp Of Philadelphia

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Adolescents and young adults (AYA, 15-24 yo) are often sidelined in treatment-related decision-making (TRDM), compromising their emerging autonomy, identity, and relationships, as well as impacting self-management and cancer outcomes. Shared TRDM (i.e., AYA having a voice in their cancer care with their caregivers and oncology clinicians), begins with initial decisions at diagnosis (e.g., selecting a treatment, clinical trial enrollment) and evolves to ongoing decisions (e.g., treatment modifications, side effects management, supportive care interventions). TRDM occurs in a cancer communication ecosystem comprised of AYA access to unvetted online information, as well as caregiver and oncology clinician partners who often engage in TRDM on behalf of the AYA. Our and others’ research demonstrates the connection between involvement in shared TRDM and short- term (information needs, decision regret, participation in curative cancer clinical trials) and longer-term (self- management skills, medication adherence) outcomes for AYA impacted by cancer. We developed and conducted a pilot trial of one of the first decision support interventions for AYA, DECIDES (AYA Deciding about Cancer Clinical Intervention: Decision Aid for Education and Support), which is a theoretically-informed, scalable interactive web-based tool. We propose a randomized controlled trial (RCT) of the next iteration--DECIDES 2.0, which will be upgraded to an enhanced interactive mobile-friendly website with accompanying text messages, a Spanish language version, and will be delivered closer to diagnosis. The trial will occur at three pediatric cancer centers with 250 AYA (15-24 yo) enrolled within 4 weeks of diagnosis. Caregivers can also opt-in. Aim 1 will test the efficacy of DECIDES 2.0. We hypothesize that, compared to Usual Care, AYA randomized to DECIDES 2.0 will report greater involvement in shared TRDM at 8 weeks post-randomization (primary outcome) and 16 weeks, more positive decision-related processes (patient activation, fewer unmet information needs, less decision regret, more decision self-efficacy, increased preference for involvement), and better self-management (skills, allocation of responsibility, medication adherence) at 8 and 16 weeks. We will explore if patient activation and/or decision self-efficacy mediates or moderates the association of TRDM with self-management. To prepare for dissemination, Aim 2 will evaluate barriers and facilitators of implementation based on the Consolidated Framework for Implementation Research. Specifically, we will assess factors contributing to DECIDES 2.0 efficacy and identify how best to integrate it into practice via quantitative + qualitative evaluation of AYA, caregivers, and oncology clinicians’ perceived acceptability and implementation barriers and facilitators, and implementation feasibility and costs. Responsive to PAR-25-167, and led by a stellar multidisciplinary team, this proposal advances efforts to modify the cancer communication ecosystem and optimize cancer care for AYA by targeting their involvement in shared TRDM, ultimately optimizing medical and psychosocial outcomes. Results will inform a future implementation trial of DECIDES 2.0 across pediatric cancer programs.

NIH Research Projects · FY 2026 · 2026-06

Abstract Aicardi-Goutières Syndrome (AGS) is a heritable interferonopathy that is now treatable by blocking the interferon receptor, IFNAR. However, AGS does not have any biomarkers to guide therapeutic decision-making. We propose to leverage our active clinical program in AGS, currently managing over 170 affected individuals, and our demonstrated ability to measure the Interferon Signaling Response (ISR) to explore a composite biomarker for this context of use. This work is enabled by a rigorous clinical data science environment in our NIH-funded consortium, the Global Leukodystrophy Initiative Clinical Trials Network (GLIA-CTN), the support of the new Center for Diagnostic Innovation (CDI) at CHOP, and a multidisciplinary investigative team including disease experts, outcome validation teams, biomarker validation research staff, patient advocates, data analysts, and biostatisticians. This team will explore the ISR as a monitoring biomarker in the context of AGS therapies, which will transform our ability to treat this rare disease population. In Aim 1, we will transition this test to the CDI, defining its analytic and clinical validity for the Context of Use (COU). In Aim 2, we will test the use of the ISR and thresholds prospectively in its COU by using it to guide therapy modification in our active clinical AGS therapy program, which treats more than a dozen newly affected subjects a year. At the conclusion of this program, we expect to be able to submit the ISR to the FDA for consideration as part of the biomarker qualification program. The overall expected outcome of this proposal is the demonstration of the ISR as fit-for-purpose as a disease monitoring biomarker in AGS therapies and readiness to advance the ISR to full clinical biomarker validation.

NIH Research Projects · FY 2026 · 2026-06

SUMMARY Irritable bowel syndrome (IBS) is a disorder of gut-brain interaction with a childhood prevalence of up to 5.1% in the United States, and up to 22.6% worldwide. Children with IBS experience debilitating psychosocial and gastrointestinal (GI) symptoms resulting in impaired quality of life (QoL). IBS with constipation (IBS-C) is a subtype characterized by abdominal pain, distension or bloating, straining, and infrequent hard stools. While this is a pervasive issue, little is known about the underlying pathophysiology of IBS-C; it is thought to relate to both bowel dysmotility as well as vagal tone dysfunction. Current treatments for IBS-C are limited and often incompletely relieve pain. Integrative health therapies have become new therapeutic avenues in the management of IBS. Myofascial release (MFR) is a form of hands-on physical therapy that targets the myofascial interspaces to increase abdominal wall tissue motility. MFR used during osteopathic manipulations improves abdominal distention, constipation, and QoL in adults with IBS and increases gastric myoelectrical activity in healthy adults as measured by electrogastrography. However, the effect of abdominal-MFR on the activity of other bowel regions is unknown. We have developed and evaluated an abdominal-MFR protocol and shown that using abdominal-MFR in adolescents with IBS is feasible and well accepted without adverse events. Preliminary studies show a significant improvement in GI symptoms and QoL up to one month after the abdominal-MFR intervention. However, the physiologic mechanisms by which MFR improves IBS symptoms have not been investigated, there is a lack of objective methods to assess these physiologic outcomes, and there are no randomized clinical trials assessing MFR's effectiveness in children. MFR therapies could offer a low risk, high- benefit treatment that can be incorporated into current treatment regimens but are understudied in IBS. Our overall objective is to evaluate the physiologic effect of abdominal MFR that results in symptom improvement in children with IBS-C using a randomized comparative trial design. In Aim 1 we will assess the effect of abdominal- MFR versus light-touch-massage on vagal tone (assessed using heart rate variability) and correlate with GI and psychosocial symptoms. We hypothesize that abdominal-MFR improves both GI and psychosocial IBS- associated symptoms by eliciting an improvement in vagal tone. In Aim 2 we will evaluate the effect of abdominal- MFR versus light-touch-massage on myoelectrical activity (gastric, small bowel, and colonic) using a non- invasive abdominal surface wireless motility monitoring system and correlate with symptoms. We hypothesize that abdominal-MFR will induce statistically significant organ specific differences in bowel activity that will correlate with a significant improvement in GI symptoms. Our innovative approach employs established and new non-invasive vagal tone and bowel activity testing strategies. This study will address several knowledge gaps surrounding the mechanisms behind MFR's improvement in IBS-C symptoms with great potential to support non- invasive evidence-based treatments in other IBS subtypes and disorders of gut-brain interaction.

NIH Research Projects · FY 2026 · 2026-06

Heterotopic ossification (HO) consists of formation of extraskeletal bone -usually endochondral in nature- at ectopic sites at the expense and damage of muscles, tendons, and other local tissues. This pathology is caused by a variety of insults that include invasive surgeries, severe trauma, and injury to the central nervous system. It is also often observed in patients who are implanted with recombinant bone morphogenetic protein 2 or 7 (rhBMP2 or rhBMP7) to treat bone deficiencies or induce vertebral fusions to ameliorate spine function and reduce pain. The formation and accumulation of HO tissue can cause various health problems including impingement of muscles, nerves, and other tissues, reduction of joint mobility, chronic pain and difficulties in prosthesis fitting. Currently, preventive treatments include systemic administration of non-steroidal anti- inflammatory drugs (NSAIDs), local low-dose irradiation or a combination of the two. Animals treated with indomethacin are more resistant to HO, but NSAID treatment can increase bleeding by affecting platelet function, exacerbate gastritis and even impede fracture repair in trauma patients. Radiation can decrease soft tissue healing and dampen beneficial immunological responses. Surgery can be used to remove the HO mass, but it can be problematic and can even trigger another round of HO. Recent animal studies have suggested possible new strategies for HO prevention, but it is unknown whether any of those has concrete clinical value. In recent preliminary studies, we have examined models of HO in adult mice that involve local tissue damage and induction of traumatic HO. In these models, the pathogenic process recapitulates human HO, starting with recruitment of local progenitor cells, commitment to chondrogenesis and eventual deposition of endochondral bone. We searched for new culprits of pathogenesis and identified a protein that appears to be central to the onset of the HO formation cascade. When we experimentally impeded the function of the protein by pharmacologic means, the onset of HO was delayed, and the ultimate amount of HO was greatly diminished. In this project, we propose testing whether genetic ablation of the gene encoding the protein will prevent HO formation in traumatic mouse HO models. We will then carry out cellular analyses to clarify the mechanisms by which the protein exerts its function. In the last aim, we will further test the efficacy and safety of the pharmacologic treatment. Procedures and approaches to be used will include transgenic mouse models, cell lineage tracing, single cell RNA transcriptomics, immunofluorescence, microCT imaging, fluorescence recovery after photobleaching (FRAP) and protein-protein interaction assays. The project is based on novel data and insights and promises to provide conclusive evidence for the role of the protein in HO pathogenesis. This in turn would solidify the notion that targeting the protein could provide an effective new treatment against HO, paving the way for future clinically relevant projects.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY For many patients with intractable epilepsy, brain surgery becomes a viable consideration to reduce the devastating consequences of seizures. Functional imaging with techniques such as magnetoencephalography (MEG) also allow delineation of “eloquent” cortex, such as anatomic areas supporting language function, to be spared during surgery. However, conventional MEG requires patients to hold still in order to get high quality functional maps, which is often challenging for pediatric patients. Further, conventional (cryogenic) MEG houses its detectors in a rigid (adult-sized) helmet, consequently leading to an increased distance between neural generators and magnetic field detectors, when used for pediatric patients. This distance leads to a loss of sensitivity (since magnetic fields fall off with the square of distance). Thus, current cryogenic MEG Is not ideally suited to the smaller, more movement-prone pediatric population. The current proposal uses an entirely new MEG technology, based on a device called an OPM (optically pumped magnetometer), which does not require cryogenic cooling and thus can be flexibly configured into a “wearable MEG” array. Since the detectors are effectively scalp-mounted, they have increased sensitivity. Since they are effectively rigidly-coupled to the head, they tolerate head motion. Combining these two attributes promises to ameliorate the above-described limitations to presurgical mapping of language function in the brains of pediatric epilepsy patients. The goal of this proposal is first to spatially validate the language map foci, compared to conventional MEG (in cases where conventional MEG is diagnostically successful) and secondly, to increase the diagnostic yield of functional brain mapping with MEG (by providing diagnostically interpretable maps in cases where conventional cryogenic MEG is equivocal or insufficient). 48 pediatric epilepsy patients (8-18years) undergoing conventional cryogenic MEG as a part of their clinical standard of care presurgical evaluation will be recruited to undergo an additional brief language mapping paradigm using the novel wearable MEG device. Statistical methods will address both spatial validation of language maps determined using wearable OPM-MEG as well as the degree to which diagnostic yield of evaluable language function maps is increased using wearable MEG compared to the current clinical, cryogenic, rigid-helmet (non-wearable) MEG standard. If successful, as anticipated, it is hoped that these data will pave the way for eventual clinical adoption of wearable MEG for presurgical functional mapping of language cortices in pediatric epilepsy patients who are candidates for neurosurgery, with concomitant improved in post-operative functional outcomes and, ultimately, quality of life.

NIH Research Projects · FY 2026 · 2026-05

We request funds to acquire a Bruker timsTOF Ultra 2 mass spectrometers for the Proteomics Core Facility at the Children’s Hospital of Philadelphia (CHOP) Research Institute. This high-end instrument will replace an aging Thermo QE HF system, which no longer meets the sensitivity, speed, or flexibility demands of modern proteomics. The Ultra 2 will dramatically expand our capacity to support NIH-funded research projects that rely on high-resolution, quantitative proteomic profiling of clinically relevant and low-input samples, many of which are not feasible with our current instrumentation. The CHOP Proteomics Core has a strong record of enabling both basic and translational research, with deep expertise in mass spectrometry method development and collaborative project support. Our team has contributed to numerous NIH-funded studies in pediatric heart disease, cancer, metabolic disorders, and infectious diseases. Following a rigorous head-to-head evaluation, the Bruker timsTOF Ultra 2 was selected for its superior sensitivity, acquisition speed, and upgradable modular design, features particularly critical for HLA peptidomics and other advanced applications. The instrument will be housed in a well-supported core facility with dedicated infrastructure and oversight by a newly established Advisory Committee to ensure equitable access and sustained scientific impact. With 15 NIH-funded investigators identified as major and minor users, and an additional 11 researchers supported by non-NIH funding or preparing NIH submissions, this acquisition will directly advance cutting-edge research and expand access to transformative proteomics technologies across the CHOP research community.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Conversation is a dynamic interplay between what you say and how you say it, including what your face and voice express. Even autistic individuals who score within the typical range on standardized language tests frequently report less successful and satisfying social communication outcomes than non-autistic peers, which may be related to differences in the integration and use of verbal and non-verbal expressions. Unfortunately, research mostly fails to consider language in its interactional and multi-modal context in two crucial ways: 1) The intra-speaker alignment of language and non-verbal information from facial and vocal expressions of each speaker, and 2) the between-speaker synchrony of verbal and non-verbal features. This represents a major impediment to our understanding of autistic communication. This project takes an innovative approach to analyzing how the integration of verbal and non-verbal cues predict communicative success in 560 autistic and non-autistic youth during peer-to-peer Zoom conversations. A key strength of our plan is to analyze novel facial and vocal behaviors in conjunction with language measures from an ongoing study of the same youth during neurotype-concordant and neurotype-discordant conversations. We propose to leverage multiple techniques, the unique combined expertise of this team, and access to a rich, video-based conversational dataset, to determine how verbal and non-verbal elements together predict self-reported conversational success and satisfaction in autistic and non-autistic adolescents. Three key strengths contribute to our project’s feasibility: 1) Expertise in manual coding of intra-speaker verbal and non-verbal conversational alignment that captures socially relevant details (MPI Grossman); 2) Expertise in computer vision-driven, between-speaker multi-modal synchrony using large scale datasets (MPI Herrington); 3) Access to conversational outcomes measures based on a video dataset of 560 autistic and non-autistic adolescents from an ongoing NIH-funded grant (R01 DC021564-01, Ready to CONNECT: Conversation and Language in Autistic Teens, Dr. Grossman, Co-I). The CONNECT grant captures videos of Zoom conversations between neurotype-matched and -discordant adolescents and includes self-reported measures of participants’ conversational satisfaction. However, the CONNECT project will analyze only verbal predictors of communicative success and satisfaction. The present proposal will exploit this rich video dataset to analyze non-verbal and verbal conversational behavior jointly. We will use cutting-edge computer vision techniques to determine the relative contributions of verbal and non-verbal behaviors, as well as derive network models of nuanced individual communication profiles that can inform more individualized intervention models. The CONNECT team supports this proposal (see letters of support) and both MPIs Herrington and Grossman have full data access and associated IRB approvals. This project represents a huge economy of scale and has the capacity to identify “subgroups and subpopulations” within autism to support individualized interventions (NIDCD’s Strategic Plan Theme 3).

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY A growing number of disease-causing genes underlying childhood epilepsies have been discovered, many of which can be targeted with precision medicines. Genetic etiologies are implicated in a wide range of childhood epilepsies, including lesional brain disorders. We have previously generated genome sequencing data in children with lesional epilepsies through the Kids First X01 mechanism. We now propose to jointly analyze genomic sequencing data with longitudinal clinical data utilizing the Fast Healthcare Interoperability Resources (FHIR) data model, which is currently implemented within the Gabriella Miller Kids First Pediatric Research Program. While the Kids First data resource offers foundational clinical information, a more extensive array of longitudinal clinical data from Electronic Medical Records (EMR) is vital to deeply comprehend the influence of genetic variation on the clinical trajectory of childhood epilepsies. Our current initiative advocates for the integration of genomic data from two Kids First Cohorts, specifically targeting epilepsy and brain tumors, with longitudinal EMR data. This strategy is designed to analyze the phenotypic trajectories associated with rare genomic variants and assess the combined predictive strength of clinical and genomic data in anticipating disease-relevant outcomes. First, we aim to outline disease trajectories and treatment landscapes linked to rare genomic variants (Aim #1). We will establish longitudinal disease histories across 13,983 patient years, including diagnosis codes, procedures, and medications using the Human Phenotype Ontology (HPO). We will assess phenotypic associations, treatments, and procedures in monthly increments for established and candidate genetic etiologies and outline clinical trajectories and treatment landscapes of individuals carrying rare variants. This analysis will provide the most comprehensive picture of underlying genotype-phenotype association to date. In addition, we will assess the predictive power of joint clinical and genomic data for disease-relevant outcomes (Aim #2). A combination of genomic information and limited EMR data is sufficient to predict severe childhood epilepsies early in the disease course. We will define 25 EMR-based outcomes and develop decision-tree and Random Forest models to identify combinations with high predictive power for each outcome, followed by assessing age- based feature importance. This analysis will provide an overview of how clinical features and rare genetic variants can jointly predict clinical outcomes in a broad cohort of children with brain lesions. At the end of this R03 project, we will have assessed both the phenotypic consequences of rare genomic variation in lesional brain disorders and the predictive power of joint clinical and genomic information. Our project will serve as a pilot, assessing how FHIR-derived, deidentified EMR data deposited in the Kids First Data Resource can be leveraged to gain insight into the trajectories of lesional brain disorders and predict clinical outcomes.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Membranous nephropathy (MN) is an autoimmune glomerular disease that represents a major cause of nephrotic syndrome in adults and can also affect children. Patients with MN have substantial disease burden, with high risks of infection, thromboembolic events, cardiovascular complications, and kidney failure. While recent discoveries of target antigens for autoantibodies have accelerated understanding of MN biology, translation to treatment guidance is lacking. There are no FDA-approved treatments for MN and limited evidence exists for optimal use of currently available immunosuppression medications, since clinical trials are limited by small sample size, short follow-up, and a focus on short-term proteinuria outcomes. The overarching goal of this project is to emulate clinical trials by applying modern causal inference methods to observational data with long follow-up to fill these gaps in knowledge and inform future trial design. Specific Aims are to (1) estimate real-world comparative treatment effects on long-term clinical effectiveness and safety outcomes; (2) evaluate the prognostic value of the anti-phospholipase A2 receptor antibody (PLA2R-Ab) as a biomarker for long-term clinical outcomes, overall and within treatment subgroups; and (3) validate PLA2R-Ab as a surrogate endpoint for long-term clinical outcomes. Data from two observational cohort studies—the Nephrotic Syndrome Study Network (NEPTUNE) and Cure Glomerulonephropathy (CureGN)—and electronic health records from two registries—the Kidney Research Network (KRN) and Glomerular Disease Collaborative Network (GDCN)—will be pooled and analyzed using longitudinal matching, mediation analyses, and longitudinal and survival data analysis methods. Results from this study will provide evidence to nephrologists and patients to guide treatment decisions and optimize use of PLA2R-Ab for patient monitoring. Validation of PLA2R-Ab as a surrogate endpoint would provide early indications of treatment response to support disease management and facilitate shorter duration of clinical trials for future drug development.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY This K08 mentored research career development award proposal details a robust training program to position me to lead an independent R01 funded research program focused on the mechanisms of neurodevelopmental disorders. I am an Instructor in the Division of Neurology at The Children's Hospital of Philadelphia (CHOP). My clinical and research interests converge at the intersection of neurogenetics, autism and epilepsy. My prior research experience in the laboratories of Dr. Rafael Yuste at Columbia University and Dr. Adam Carter at New York University built my expertise in anatomical analysis, brain slice electrophysiology and optogenetics focused on the study of GABAergic interneurons and cell-type specificity of neural circuit connectivity. This proposal will advance my scientific training in cellular and circuit neuroscience to include animal models of neurodevelopmental disorders, in vivo two photon imaging integrated with head-fixed behavior, advanced genetic techniques and machine learning analysis of spontaneous behavior. I will be mentored by Dr. Ethan Goldberg, Associate Professor of Neurology and Neuroscience at CHOP, and Dr. Marc Fuccillo, Associate Professor of Neuroscience at The University of Pennsylvania (UPenn) who have co-mentored my NIH NINDS R25 fellowship. Dr. Goldberg's expertise in animal models of epilepsy, electrophysiology, and 2-photon calcium imaging complements Dr. Fuccillo's expertise in animal models of autism, striatal circuitry, quantitative behavioral analyses and in vivo neural recordings. Additionally, Dr. Goldberg is a highly successful physician scientist himself who was a prior K08 awardee who will provide invaluable career mentorship. I will receive additional mentorship and intellectual development from my advisory committee and the vibrant neuroscience and autism research communities at CHOP and UPenn. This proposal aims to identify mechanisms of autism in a mouse model of Dravet Syndrome (DS), a genetic neurodevelopmental disorder presenting with infantile onset epilepsy and high rates of autism. I will uncover the cellular pathophysiology of DS in subtype-specific striatal interneurons (INs) and investigate how this impacts excitation-inhibition balance in the striatum. I will then use in vivo 2P imaging in the striatum to reveal the impact in different cell types on neuronal activity during distinct behavioral states and neural encoding of goal-directed behavior. This will be the first use of in vivo 2P imaging in the striatum in a disease model. Finally, I will identify behavioral correlates of striatal IN dysfunction and test the contribution of distinct striatal IN subtypes to behavior using a novel intersectional genetic approach to selective gene loss or re-expression. Together training in these cutting-edge approaches combined with the exemplary resources available to me at CHOP and UPenn will prepare me for independence as a physician scientist making crucial discoveries to improve our understanding and therapeutic approaches to neurodevelopmental disorders.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Adoptive cell therapy with chimeric antigen receptor (CAR) T cells targeting a single tumor-specific surface antigen has demonstrated remarkable long-term efficacy in certain hematological malignancies. This success led to its approval by the FDA for the treatment of multiple hematological cancers. However, the clinical outcomes of CAR T therapy in solid tumors have been largely disappointing. A key obstacle is the pre-existing antigen heterogeneity in solid tumors, which often results in antigen-negative tumor escape, because not all tumor cells express the antigen targeted by the CAR. Pre-clinical and clinical studies have shown that polyclonal T-cell responses against different tumor antigens, or even the same antigen, could lead to superior tumor control in the long term. We recently published that vaccine boosting of CAR T cells stimulated CAR T cell expansion with enhanced functionality and unexpectedly triggered antigen spreading, resulting in more effective treatment of tumors with antigen heterogeneity in an animal model. More specifically, this was achieved by using a lymph- node targeting lipid polymer to deliver the natural ligand recognized by the CAR to antigen-presenting cells (APCs). However, the kinetics and magnitude of antigen spreading may not be sufficient for treating tumors with a high proportion of tumor cells missing the CAR antigen. Building on these findings, we will develop an autologous tumor cell membrane-coated dual CAR and endogenous T cell boosting virus-like nanoparticle vaccine (dtVLP-vax) to overcome this barrier. The dtVLP-vax particles retain on their surfaces a collection of tumor-associated antigens (TAAs), including the antigen recognized by the CAR and peptide-major histocompatibility complexes (pMHCs) recognized by T cell receptors (TCRs). These antigens will be transferred onto the APCs upon arrival in the LN, allowing natural APCs to boost both adoptively transferred tumor-targeting CAR T cells and endogenous tumor-specific T cells, generating a polyclonal anti-tumor T cell response. The unbiased presentation of all TAAs, especially pMHCs, by dtVLP-vax also eliminates the need for tumor sequencing and neoantigen identification, which is often costly and time-consuming. We hypothesize that dtVLP-vax can efficiently support CAR T therapy and amplify endogenous T cells to rapidly diversify and broaden the anti-tumor T cell immunity, thereby facilitating a more effective control of solid tumors with antigen heterogeneity. Using multiple solid tumor models, we aim to (1) Characterize the biology and mechanisms of dtVLP-vax for stimulating CAR T and endogenous T cells in vivo. (2) Assess the versatility of the dtVLP-vax platform for reprogramming T cells. (3) Evaluate the capacity of dtVLP-vax to boost both CAR T cells and endogenous T cells with enhanced diversity and functionality against solid tumors displaying antigen heterogeneity. (4) Assess the human tumor-derived dtVLP-vax. These studies will establish key principles of dtVLP-vax designs to maximize the potential of adoptive cell therapy for solid tumors.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY / ABSTRACT The developmental switch from fetal (HbF) to adult (HbA) hemoglobin expression is important scientifically as a paradigm of developmental transcriptional control and clinically as a target for treatment of hemoglobinopathies. Reversing this switch to increase HbF expression is an essential approach for gene therapy and new small molecule therapeutics to treat sickle cell disease and b-thalassemia. BCL11A, expressed exclusively in adult cells, is the predominant transcriptional repressor of HbF. However, attempts to develop specific small molecule BCL11A modulators have not been successful, leading to a concerted effort to understand the developmental regulation of BCL11A itself to uncover basic principles of developmental globin control and identify more amenable targets. We used two converging approaches to identify the CHARGE- syndrome associated chromatin remodeler CHD7 as a specific HbF regulator. First, CHD7 scored as an HbF activator in a CRISPR screen of chromatin remodeling proteins. Independently, we found that CHD7 is expressed at higher levels in a small subset of adult erythroblasts that can turn on HbF either spontaneously or in response to pharmacological inducers. In preliminary experiments, we find that CHD7 is expressed predominantly in fetal cells, but that ablation of residual CHD7 in adult erythroblasts leads to transcriptional upregulation of BCL11A and repression of HbF. We hypothesize that CHD7 is a developmentally controlled activator of HbF expression through BCL11A and potentially through direct action at the b-globin gene locus. In Aim 1, we will determine the mechanisms of CHD7 gene selectivity in globin gene regulation. In Aim 2, we will characterize the developmental control of CHD7 expression. This dissection of CHD7’s regulation and selectivity may provide new opportunities to therapeutically manipulate fetal hemoglobin for the treatment of sickle cell disease (SCD) and b-thalassemias. Additionally, we will use the extensive tools and data sets developed in erythroid cells to better understand the activity of CHD7 in other biological contexts.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract The objective of the Children's Hospital of Philadelphia (CHOP) Research Institute Summer Scholars Program (CRISSP) is to provide undergraduates with advanced training and skill development required for future leaders in basic, translational, clinical, and behavioral child health research. We will identify and nurture the next generation of innovators in pediatric research. Undergraduate scholars are often at a professional inflection point where the targeted efforts of CRISSP are ideally timed to provide skills, knowledge of scientific career options, and mentors dedicated to pioneering advances that ensure children lead healthy and productive lives, free from disease or disability, from birth to adulthood. The proposed research education program is designed to improve on an existing outstanding 10-week summer pediatric research internship with the ultimate goal of creating uniquely qualified students who are primed to both succeed in and foster the biomedical research workforce path. The annual program, which provides mentored research training to 25-30 competitively chosen undergraduate interns, will achieve this goal through three synergistic strategies: 1) scientific mastery through completion of an independent research project, 2) career exploration through shadowing and active mentorship, and 3) academic skill development through tailored curriculum. A dedicated team of mentors and program staff support CRISSP scholars both during and after the program, engaging alumni to both contribute to the experiences of current CRISSP scholars and to serve as peer and professional networks to help advance career development. CRISSP is creating an exceptional cadre of young scientist- leaders, gifted with an appreciation aligned with the mission of the National Institute of Health to transform discoveries into more healthy lives.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT The K23 application proposes to examine learning and neurocognitive mechanisms underlying food approach in avoidant/restrictive food intake disorder (ARFID) using computational methods and will position the applicant, Marita Cooper, Ph.D., to transition to research independence with expertise using computational modeling to examine mechanisms of restrictive eating disorders (ED) in youth. ARFID is the most prevalent ED in childhood, with sequelae including malnutrition, delayed growth, cardiac complications, and death. Early data suggest youth with ARFID exhibit executive functioning deficits, including weak central coherence and poor response inhibition. These data underly the hypothesis that youth with ARFID may be slow to learn food approach, impacting food intake, and requiring more exposure to novel foods for learning to occur. Knowledge of neurocognition and learning in ARFID is in its infancy, yet computational modeling offers an innovative approach to probe underlying processes and identify target mechanisms related to aberrant food approach. Two approaches with utility in other EDs, active inference and reinforcement learning, have not been applied to ARFID. The study will examine learning mechanisms and neurocognition of aberrant food approach in ARFID. We will recruit 99 youth (66 with ARFID, 33 controls) ages 8-18, matched on age and sex. Participants will complete a three-armed bandit task, assessing learning mechanisms (via food and neutral stimuli), and a meal-based buffet task assessing food approach (macronutrient and caloric intake). We will assess neurocognition, ED symptoms, and approach/ avoidance. Aim 1 hypothesizes that youth with ARFID will exhibit poorer performance (under both neutral and food conditions) than healthy controls and that worse performance will relate to overall intake during the buffet task. Aim 2 follows participants naturalistically, repeating assessments at 6- and 12-month follow-up. We will examine whether baseline performance predicts improvement in ARFID symptoms at follow-up. Aim 3 will compare whether active inference or reinforcement learning models best fit participant learning behavior. The project will be an important major step in developing a data-driven model of ARFID, providing critical information about potential drivers of aberrant food approach. The proposed project will support expert mentorship and training for Dr. Cooper including 1) learning and neurocognitive development in youth; 2) conducting and managing longitudinal research in clinical samples; and 3) practical skills in computational modeling transferrable to future research. The resources of Children’s Hospital of Philadelphia and University of Pennsylvania and an expert team of mentors (with expertise in mechanisms of ARFID/restrictive ED, development, clinical research, and computational modeling) provide an outstanding context to launch Dr. Cooper’s career. Project findings are consistent with the NIMH strategic goal to identify validated targets for intervention and will inform a competitive R01 application examining computational learning mechanisms in a transdiagnostic sample of youth with restrictive ED.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT BLOC1S1-related leukoencephalopathy (BLOC1S1-LE) is a rare neurological condition characterized by hypomyelination, epilepsy, and global developmental delays. BLOC1S1 is a shared subunit of the Biogenesis of Lysosomal Organelles Complex-1(BLOC1) and BLOC-One-Related Complex (BORC) that govern various endo- lysosomal (LYS) pathways. BORC mediates kinesin-dependent transport of LYS toward autophagosomes (with Arl8-SKIP-kinesin) and conducts homotypic fusion and protein sorting (HOPS)-supported autophagosome-LYS and endosome-LYS fusion. Although several subunits of BORC result in severe myelin deficits, how BORC causes myelin deficits remains unclear. This proposal aims to uncover how the BORC-HOPS complex dysregulates autophagy and affects the differentiation of oligodendrocyte precursor cells (OPCs) to oligodendrocytes (OLs; myelin-producing cells). To address this, we will use conditional mouse models targeting OPCs and assess whether Bloc1s1 depletion disrupts the BORC-HOPS- N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) ensemble, henceforth disrupting the autophagosome-LYS fusion and dysregulating autophagy. We will also generate OPC and OL-like cells from patient-derived iPSC of BLOC1S1- LE and replicate these findings (Aim 1). As OLs are the sole producers of myelin in the central nervous system, we will conditionally delete Bloc1s1 in OLs and assess the transport of myelin proteins and cholesterol, essential components for myelin formation. We will also evaluate if alterations in BORC disrupt LYS biogenesis and transport of key myelin components (Aim 2). To confirm the functional relation of LYS deficits to BLOC1S1-loss, we will attempt to rescue OL lineages with a lentiviral construct of BLOC1S1 in murine and iPSC-derived OPC and OLs. This proposal will address a gap in knowledge on how LYS function and vesicle homeostasis are essential for myelin formation.

NIH Research Projects · FY 2026 · 2026-04

Abstract This project focuses on macromolecular prodrug-based delivery of a topoisomerase I inhibitor, SN22, structurally enhanced to overcome tumor defense mechanisms in order to achieve durable suppression of high-risk neuroblastoma (NB), the most common and deadly solid tumor of childhood. The intensive, multimodality treatment currently used clinically fails in over half of high-risk NB patients: 50-60% experience a relapse with no curative salvage treatment options. Centered on developing and optimizing a drug delivery strategy against the aggressive disease not responding to conventional therapies, with a particular focus on a high-risk form of multiple drug-resistant NB with increased “stemness” driven by a MYCN protooncogene and its downstream target, ABCG2 (an ABC drug efflux pump suppressing chemosensitivity and promoting tumorigenicity), this project will evaluate an approach integrating polymer-linked prodrug design and structural optimization of the cargo to improve delivery, extend drug residence in the tumor, and reverse drug resistance. Guided by our past work and the results of our preliminary studies toward this project, we hypothesize that prodrug-mediated delivery of SN22 will potently suppress growth of aggressive, pre-therapy and chemorelapsed NB tumors by enhancing drug uptake and extending tumor exposure to therapeutically effective drug levels and by taking advantage of the inactivation-resistant molecular design of this agent. This hypothesis will be tested by pursuing the following specific aims: Aim 1 studies will focus on in vitro evaluation of a series of prodrug constructs on NB cells derived at relapse from MYCN-amplified high-risk NB tumors; Aim 2 studies will examine tumor uptake, biodistribution and elimination of the prodrugs in orthotopic xenograft NB models; Aim 3 experiments will comparatively evaluate antitumor efficacy of prodrug- mediated delivery in models of newly diagnosed and recurrent MYCN-amplified NB in comparison to a new syngenetic model of disseminated (MYCN-driven) high-risk disease. Through optimizing the design and performance of SN22 prodrugs using a panel of clinically relevant models recapitulating distinct types and phases of aggressive NB, this research is expected to have a strong impact on the field by addressing several barriers to the translation and clinical implementation of macromolecular prodrugs and by paving the way to improved clinical management of drug-resistant NB and other high- risk cancers showing minimal or no response to conventional therapies and currently lacking effective treatment options.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY/ABSTRACT Technologies used to manage type 1 diabetes (T1D) are advancing rapidly, particularly automated insulin delivery (AID) systems. AID uses real-time continuous glucose monitoring (CGM) data and a controller algorithm to automate subcutaneous insulin delivery through an insulin pump. Despite the clear clinical benefits, AID use declines over time, and children and adolescents are more likely to discontinue AID compared to adults. The development and evaluation of interventions to promote AID use is critical to increase long-term use of AID and improve T1D health outcomes. The decision making process related to choosing and implementing AID- especially the youth’s involvement in the decision- may lay the groundwork for effective AID use over time. Prior research has demonstrated that youth decision making involvement (DMI) is associated with T1D adherence and technology use. Therefore, we developed components for a parent- and youth- directed intervention, implemented by trained research staff (i.e., “health coaches”), to promote youth DMI in decision making related to choosing and implementing AID, called Partnering with Adolescents to Choose and Implement Technology in the Management of T1D (PAC-IT). Intervention components are based on social cognitive learning theory and include education, goal setting, behavioral prompts, and structured parent- adolescent discussions about AID. The objective of this proposal is to refine and pilot test PAC-IT, with the long-term goal of facilitating consistent AID use in youth over time. This proposal is in response to PAS-25-102, “Small R01s for Clinical Trials Targeting Diseases within the Mission of NIDDK,” which does not require preliminary data. Aim 1 is to assess acceptability, barriers, and facilitators of components of PAC-IT to promote youth DMI in decision making about AID, using semi-structured interviews. Aims 2-3 are to evaluate the feasibility and acceptability of PAC-IT and determine if PAC-IT impacts AID satisfaction, AID self-efficacy, sustained AID use, and glycemic control in a 1:1 randomized controlled trial, comparing PAC-IT to attention control. Outcomes will be assessed immediately post-intervention and at 6-month follow-up (approximately 8.5 months after AID implementation. At the conclusion of this study, we will be ready to evaluate PAC-IT in a larger, fully powered trial in a sample of youth with T1D, with the ultimate goal of maximizing the clinical benefits that can be obtained by effective use of this important technology.

NIH Research Projects · FY 2026 · 2026-03

Project Summary Marijuana (MJ) use among people living with HIV (PWH) is up to three times more prevalent than in the general population. MJ can be consumed by ingesting cannabis (e.g., edibles like pills) or smoking (i.e., inhaling MJ smoke), providing benefits to PWH, such as reduced stress, anxiety, pain, and depression. The impact of MJ edibles on the cognitive function of PWH has not been explored. However, MJ smoke contains many of the same toxins and carcinogens as tobacco (TC) smoke, which may increase oxidative stress and negatively affect the antioxidant glutathione levels (GSH – a marker of oxidative stress) in the brain of PWH. Advanced edited magnetic resonance spectroscopy (MRS) can measure GSH in PWH who smoke MJ and provide insights into how MJ smoking contributes to oxidative stress and resulting cognitive impairment. Furthermore, the toxins in MJ smoke elevate the risk of inflammation and plaque formation in blood vessels, causing vasoconstriction or vasodilation, which alters cerebral blood flow (CBF). These mechanisms leading to adverse hemodynamic perturbations may be more pronounced in PWH who smoke MJ than in those who consume MJ edibles. Alterations in CBF may result in insufficient oxygen for mitochondria needed to synthesize adenosine triphosphate (ATP)-dependent GSH for antioxidant protection against reactive oxygen species (ROS), leaving PWH smokers vulnerable to oxidative stress and neuronal damage. In this proposal, we will assess brain GSH and CBF in PWH who use MJ (smoking vs. edibles). We will also compare the effects of MJ smoke with TC smoke on GSH levels in PWH. In Aim 1, we will evaluate GSH profiles in five groups of individuals using advanced edited MRS: PWH MJ smokers, PWH who use MJ edibles, PWH TC smokers, PWH who do not smoke TC and do not smoke MJ or take MJ edibles (TC–/MJ–), and seronegative (SN) TC–/MJ–. We hypothesize that both PWH MJ smokers and PWH TC smokers will exhibit similar GSH levels, though these levels will be the lowest among all groups, indicating that smoking leads to an overproduction of ROS and increased oxidative stress. PWH who consume MJ edibles and PWH TC–/MJ– will have comparable GSH levels, suggesting that edibles do not induce oxidative stress, while PWH TC–/MJ– will display lower GSH than SN TC–/MJ–, indicating ongoing oxidative stress due to HIV. In Aim 2, we will evaluate regional CBF in PWH who smoke MJ and those who use MJ edibles. We hypothesize that PWH who smoke MJ will demonstrate impaired CBF (the lowest), indicating ROS-mediated vascular changes related to smoking, which will correlate with lower GSH levels. This proposal aligns with NIDA’s agenda to develop targeted mitochondrial function and dynamics indicators, including oxidative stress. Measurements of GSH and cerebral blood flow will offer insights into the pathophysiology linked to HIV infection and MJ use, potentially paving the way for new therapeutic strategies addressing oxidative stress and neuronal injury.

NIH Research Projects · FY 2026 · 2026-02

Kingella kingae is an invasive gram-negative pathogen that is a leading cause of bone and joint infections in young children, accounting for up to 88% of osteoarticular cases in children <4 years old. In addition, K. kingae is an important cause of invasive bloodstream infections in young children. Complications of osteoarticular infections in children include abnormal bone growth, decreased joint mobility, unstable joint articulation, and chronic joint dislocation, with residual skeletal dysfunction in 10-25% of cases. Complications of invasive bloodstream infections include multi-organ injury and mortality. Roughly 25% of K. kingae isolates possess β-lactamase activity, and many of these isolates are resistant to other antibiotics as well, raising concern about approaches to treatment in the future and underscoring the need for novel therapeutics. The pathogenesis of K. kingae disease begins with colonization of the oropharynx, followed by invasion of the bloodstream and spread to bones, joints, and other sites. We have established that K. kingae produces type IV pili (T4P), which play a critical role in adherence to epithelial cells and extracellular matrix proteins and augment the RtxA toxin in promoting efficient translocation across polarized epithelial monolayers, important steps in colonization and invasion of the bloodstream. We have discovered that K. kingae T4P pili contain pilus-associated adhesins called PilC1 and PilC2, which promote pilus biogenesis and mediate adherence and twitching motility. PilC1 and PilC2 share some structural homology but have little amino acid homology and possess different binding and twitching motility properties. Based on mass spectrometry analysis of purified T4P, homology analysis, and structural modeling, we have found that K. kingae T4P also contain multiple minor pilins, including the FimT, PilV, PilW, PilX, and PilE minor pilins encoded by the fimTpilVWXE locus. Bacterial 2-hybrid results and AlphaFold3 predictions suggest that PilC1 and PilC2 form a heterodimer and interact with a minor pilin complex. In this proposal, deploying high- resolution electron and fluorescent microscopy, biochemical techniques, and molecular and cell biological methods, we will elucidate the physical relationship of PilC1 and PilC2 with each other, characterize the physical relationship of PilC1-PilC2 and with the pilus fiber, and define the roles of PilC1 and PilC2 in K. kingae trafficking across polarized respiratory epithelial cells. The proposed studies will provide insights into the design of novel therapeutics against K. kingae and potentially other pathogens that produce T4P and may also shed light on other closely related systems such as the type II secretion system in gram-negative bacteria, the competence system for DNA uptake in gram-positive bacteria, and flagella in archaea.

NIH Research Projects · FY 2026 · 2026-02

Abstract The central question in the urologic management of patients with spina bifida is how to best identify the subset of patients whose bladder dysfunction places their upper urinary tract at risk of developing chronic kidney disease. The current “gold standard” for assessment and risk stratification of lower urinary tract physiology is the videourodynamic study (VUDS) which has some limitations. Our preliminary pilot data suggests that a select urine peptide profile will be able to identify those hostile bladders which increase the risk of CKD progression. Our two specific aims leverage three established resources at the Children's Hospital of Philadelphia: i) a proteomics core lab with experience in urinary peptidomics, ii) an established urine biobank linked to known curated VUDS outcomes stored in a database (800 samples from ages range from 2 months to 45 years), and iii) an established machine learning algorithm to assign risk classes to VUDS data stored in our digital data repository. In specific aim 1 we will measure an array of peptides in urine samples from patients with known VUDS outcomes serving as ground truth to optimize and refine a panel of peptides to discriminate high or low urodynamic risk in a larger population. The panel will then be validated in an independent set of test samples. In specific aim 2, we will measure the predictive power of the urinary peptidome by comparing the starting low risk peptidomes of two cohorts of patients; those whose VUDS remained low risk and those whose VUDS progressed to high risk. Aim 2 tests our hypotheses that urinary peptide markers can serve as predictors of future urodynamic deterioration. The overarching goal of this project is to identify a panel of urinary peptides that serve to discriminate the high risk urodynamic patterns that place these patients at risk of impending upper tract deterioration. This would change clinical practice by allowing patients to collect a urine sample in an office setting (or preferably at home to send in by overnight courier) for peptidomic analysis and risk stratification leading to a more efficient use of clinical resources.

NIH Research Projects · FY 2026 · 2026-02

PROJECT SUMMARY The goal of this proposal is to define a novel regulatory pathway in type 2 lymphocytes that prevents chronic allergic lung inflammation and airways hyperresponsiveness. Asthma is a chronic inflammatory disease of the airways that affects 26 million Americans and results in approximately 3600 deaths per year in the US alone. Over the past 40 years, the incidence of asthma has increased across the US2. The underlying pathology of asthma is complex, and in many instances poorly understood. New therapies are being introduced that take advantage of our understanding of the importance of type II cytokines (IL-4, IL-5, IL-13). We have identified a regulatory pathway in innate type 2 lymphocytes (ILC2s) that restricts cytokine receptor signaling and thus limits ILC2 numbers and cytokine production following helminth or fungal allergen exposure. This regulatory circuit is under the control of the E3 ubiquitin ligase Cul5 and its SOCS-box containing substrate receptors. Our preliminary data demonstrate that Cul5 expression in ILC2s protects lungs from allergic inflammation. Mice lacking Cul5 in ILC2s showed increased numbers and function of lung IL-33R+ ILC2s, increased sub-epithelial fibrosis, and increased airways hyperresponsiveness following fungal allergen exposure. Interestingly, we also identified a significant increase in the numbers of atypical IL-18R+ ILC2s in the lungs. These are ILC2s more commonly seen in the skin. Intranasal administration of rIL-33 or rIL-18 resulted in increased numbers of Cul5- deficient lung ILC2 cells. This was particularly surprising given that Cul5 is largely thought to limit Jak/STAT signaling and IL-18 and IL-33 receptors are IL-1 family receptors that signal thru MYD88/NFKB. Based on our preliminary data, we posit that Cul5 works with one or more SOCS-box containing substrate receptors to limit signaling downstream of IL-18R and IL-33R, expansions of lung ILC2s, and decrease susceptibility to allergic inflammation in the lung. The studies proposed here will help us identify ways to strategically target Cul5 for therapeutic benefit. Furthermore, data from these studies will provide key information regarding how Cul5 limits type 2 responses, and how it regulates cytokine signaling in IL-18R+ and IL-33R+ ILC2s in the lung.

NIH Research Projects · FY 2026 · 2026-02

PROJECT SUMMARY/ABSTRACT Hirschsprung disease associated enterocolitis (HAEC) is the leading cause of death in children who lack enteric neurons in distal bowel, a birth defect called Hirschsprung disease. The etiology of HAEC is not well understood, but hypothesized disease mechanisms include altered gut microbes (“dysbiosis”), abnormal mucosal immune system and epithelial barrier defects. To date, there are no immune-targeted therapies to treat or prevent HAEC, but new treatments are needed. This proposal builds on the candidate’s preliminary data suggesting interleukin 22 (IL22) critically modulates HAEC risk and HAEC severity. The central hypothesis is that enteric nervous system (ENS) signaling induces IL22 release and facilitates IL22 epithelial responses to enhance mucosal immunity and strengthen epithelial barrier functions that prevent enterocolitis. The Piebald lethal (sl/sl) Hirschsprung disease mouse model of HAEC will be used, as survival of sl/sl mice is dramatically (> 3-fold) altered by diet (Tjaden et al, in BioRxiv and submitted) and IL22 mRNA is much higher in sl/sl fed a Protective diet that extends median survival (“late onset HAEC”). Aim 1 will define the cellular source(s) of IL22 from bowel regions of sl/sl model mice that develop early or late onset HAEC. In parallel, this aim tests the hypothesis that IL22 prevents HAEC, by using genetic and pharmacologic strategies to alter IL22 levels. Aim 2 will precisely define the role of IL22 on epithelial integrity, stem cell renewal and differentiation in organoids derived from sl/sl mice with early or late onset HAEC and from children with Hirschsprung disease with or without HAEC. Organoids facilitate studies of epithelial stem cell biology and IL22-epithelium interactions in the absence of microbes, neurons, or diffusible small molecules such as neurotransmitters. Collectively, these studies will determine cellular sources of IL22, the effect of ENS cells on IL22 secretion, the role of IL22 in enterocolitis, and the impact of Hirschsprung disease associated aganglionosis on epithelial cell biology. These studies build on the candidate’s training as a pediatric gastroenterologist, who has clinical exposure to the diagnosis and treatment of children with Hirschsprung disease and HAEC, as well as her basic science training in enteric nervous system biology. As the work proceeds, she will become an expert in mucosal immunology and epithelial biology with a focus on neuro-immune and neuro-epithelial interactions. The mentors, Dr. Robert Heuckeroth, and Dr. Kathryn Hamilton are experts in ENS biology and epithelial biology respectively. Both mentors have a strong commitment to mentorship and NIH funding track records. Experiments will be conducted at the Children’s Hospital of Philadelphia and Perelman School of Medicine at the University of Pennsylvania, a collegial, collaborative and state-of-the art institution. The professional development and training plan will position the candidate as a successful pediatrician-scientist, who is focused on the prevention and treatment of Hirschsprung associated enterocolitis. These studies should determine if IL22-based therapies would likely be successful in HAEC, and if a human clinical trial is appropriate.

NIH Research Projects · FY 2026 · 2026-02

Project Summary/Abstract This proposal will investigate the dynamic 3D folding of the genome at ultra-high resolution across length and time scales as it relates to gene activity. Studies in Aim 1 will investigate how newly discovered “microcompartments” between active cis-regulatory elements (CREs) form as cells exit mitosis and enter G1 phase. Because microcompartments cannot be resolved with conventional Hi-C or Micro-C methods, we will apply ultra-high resolution Region Capture Micro-C (RCMC). Using acute degradation technology, we will investigate the role of a battery of key proteins in microcompartment formation. Studies at the mitosis-to-G1 transition will be complemented using a dynamic cell transition system that considers CRE dynamics during transcriptional repression. Genome-wide insights will be obtained using newly developed machine learning- based imputation. Aim 2 is motivated by the hypothesis that similar to transcriptionally active microcompartments, intricate fine scale chromatin organization exists within heterochromatin. We will apply RCMC to representative regions of constitutive and facultative heterochromatin. These studies will be complemented by experiments perturbing key heterochromatic regulators. Furthermore, machine learning based imputation will be applied to obtain genome-wide insights, followed by validation experiments, aimed at uncovering the fine-scale microstructure of the repressive chromatin compartments. Importantly, both aims will be enhanced by mechanistic 3D polymer modeling with the goal of developing a polymer model from first principles that can explain the data, to make experimentally testable predictions, and to estimate key parameters which are otherwise not experimentally observable. In Aim 3 we will validate and extend our findings using super-resolution chromosome tracing experiments as an orthogonal and sequencing independent method. This will address if microcompartments form through simultaneous multi-way interactions, will reveal cellular or allelic heterogeneity in chromosomal folding, and deepen our understanding of the relationship between 3D genome folding and transcription.

NIH Research Projects · FY 2025 · 2026-01

SUMMARY Temporomandibular joint (TMJ) osteoarthritis (OA) is characterized by degeneration of the condylar cartilage. No effective therapies exist for restoring the damaged cartilage to accommodate pediatric patients' craniomaxillofacial growth. Regenerative therapies are needed to repair TMJ cartilage. Microfracture is a technique where perforations are created through the subchondral bone to allow for bone marrow mesenchymal stem cell (MSC) infiltration to create fibrocartilaginous repair tissue. However, MSCs are difficult to localize to the damaged area without a scaffold, and their phenotype is prone to calcification. Matrix-autologous chondrocyte implantation (MACI) is the most effective knee repair approach, where autologous knee chondrocytes are seeded on a collagen scaffold and then implanted in the defect. However, MACI is costly, requires two invasive procedures, and may not be applicable to the narrow TMJ for chondrocyte extraction. Additionally, the collagenous scaffolds that are used for knee repairs do not recapitulate the native TMJ extracellular matrix environment. The objectives of this F32 training grant are to develop regenerative approaches to heal damaged TMJ cartilage for pediatric patients and train me in these approaches to complement my tissue engineering background. Using fibro-elastic cartilage of porcine meniscus, we will use our Meniscal Decellularized (MEND) scaffold to help localize and inform progenitor cells, such as ear cartilage progenitor cells (eCPCs) or MSCs. We hypothesize that regenerative therapy adapting microfracture and MACI/matrix-induced chondrogenesis can be used to repair the damaged TMJ condylar cartilage. We will first compare the in vitro chondrogenic potential and phenotypic stability of eCPCs and MSCs in MEND. Outcomes will include biochemical assays, mechanical testing, immunohistochemistry, histology, and gene expression. Then, to assess in vivo phenotypic stability of the constructs, we will subcutaneously implant cell-seeded MEND in immunocompromised mice and analyze outcomes via biochemical assays, immunohistochemistry, histology, and gene expression. Next, we will compare the repair of adapted microfracture (simulated by an MSC injection) and MACI (simulated by empty or eCPC MEND) for porcine and human TMJ condylar cartilage regeneration in vivo using the semi-orthotopic mouse model and analyze outcomes via biochemical assays, immunohistochemistry, histology, and gene expression. These studies will uncover whether microfracture regenerates TMJ condylar cartilage and if a scaffold, potentially cellular, is needed to improve condylar regeneration. Results will inform development of regenerative therapies for degenerated TMJ condylar cartilage and produce preliminary data for a K99 application to support my training as an independent researcher in the TMJ oral and craniofacial research field.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT This collaboration between the Children’s Hospital of Philadelphia (CHOP) and the University of Pennsylvania (Penn) focuses exactly on the key priorities set out for Autism Data Science Initiative: 1) to identify genetic and environmental factors that lead to an autism diagnosis and 2) determine how these factors have either contributed to or are reflected in the rising prevalence. Most of the existing research on environmental contributions to ASD focuses on isolated exposures, neglecting the systemic nature of real-world environmental interactions. This gap limits the development of actionable risk models for early diagnosis and personalized intervention. By integrating multi-level exposome data (geocoded structural factors, individual social determinants, and perinatal exposures) with genomic and omic profiles, this study will advance understanding of how environmental contexts interact with biological susceptibility to shape ASD phenotypes, offering a foundation for tailored clinical management and novel preventive strategies. While publicly available research datasets are critical to this effort, an integrated database on a well- characterized, large, clinical population is essential. CHOP successfully implemented universal early autism screening in its primary care network in 2011, and has studied early screening, diagnostic outcomes, and prevalence through electronic health record (EHR) data since then. This cohort now includes 104,405 children born between 2008-2017 who were screened for autism at an 18- or 24-month well-child visit and had at least one additional visit within the CHOP network at 4+ years of age. Thus, this is a well- characterized and research-ready dataset of ~4000 children with autism and ~100,000 without. Our proposed three-year project is to aggregate clinical and genomic data from CHOP’s EHR, clinical and research biorepositories; with Penn’s EHR data on pregnancy, maternal, and birth outcomes, and research databases (ROA Task 1). We will also generate geo-coded exposomic data including daily air and water components, greenspace, built environment, and the Childhood Opportunity Index during pregnancy, birth, and childhood (ROA Task 2). We will then use advanced data science methods to generate hypotheses about relationships among and between variables to predict autism diagnosis and explain the increased prevalence over time (ROA Task 3). With this resource and the predictive power of our machine learning analytic plan, this project will have the statistical power to identify potential causes of autism in specific populations (e.g., starting with genetic and phenotypic subgroups). We will develop a prediction model that incorporates child-, family-, neighborhood- and community-level clinical, genomic, and exposomic data to predict autism in our cohort of 104,405 children who were all screened for autism as toddlers and who are now age 8-17 years. We will also identify genomic, exposomic, clinical-practice factors, and their interactions, that contribute to the increase in autism prevalence and heterogeneity of the autism phenotype, including increases in medical and genetic risks; changes in environmental exposures; and critical changes in diagnostic practice, criteria, and service availability. Our multi-disciplinary team represents the best clinical, informatics, genetics, data science, and community engagement expertise for developing machine learning methods and tools for large-scale, high-resolution, meaningful, and actionable autism research. If successfully implemented, this study will create an unprecedented data resource for researchers to derive specific, testable causal hypotheses and definitively answer questions regarding causes of autism in a way that was not previously possible. Furthermore, the results will parse the variance across broad domains of genomic and exposomic factors that may cause autism, which is critical for setting future research priorities.