Indiana University Indianapolis

NIH Research Projects · FY 2025 · 2025-08

Project Summary Pain in sickle cell disease (SCD) is a major lifelong complication that disproportionately affects Black and African American people. Clinically, SCD presents in two distinct phases: the steady-state phase associated with chronic pain, and the vaso-occlusive crisis (VOC) phase characterized by severe pain which requires high doses of opioids and is associated with increased risk of overdose and death, opioid-induced hyperalgesia, and lowered quality of life (QoL). Mechanistic studies directly addressing the pathophysiological mechanisms of transition from steady-state to VOC are notably limited. Recent studies suggest that the progression of VOC is linked to abnormal vasoconstriction resulting from dysfunction of the autonomic nervous system (ANS). There is an urgent need to gain a fundamental understanding of the pathophysiology underlying the transition from pain to VOC and to develop evidence-based, effective, and safe therapies for managing pain in SCD patients. Preliminary results from our completed trial (ClinicalTrials.gov Identifier: NCT05045820) suggest that verum acupuncture treatment improved patient-reported pain, QoL, and reduce opioid requirements and the number of patientreported VOC. However, our understanding of the pathophysiology of SCD and VOC, and the mechanisms by which acupuncture ameliorates clinical symptoms of SCD, remains very limited. Our recent findings revealed a complex interplay among dysregulated inflammatory mediators and autoantibodies, and their relationship with VOC, pain, and sensory sensitivity at baseline in SCD patients. Subsequently, we observed that the reduced inflammatory markers and improvements of VOC/QoL/opioid requirement outcomes significantly differ between the verum and sham treatment groups. Leveraging a new clinical trial, we propose to specifically examine the inflammatory/immune biomarkers related with the onset and progression of VOC and their modulation by acupuncture through three Aims. Aim 1: Characterize variation in inflammatory/immune profiles in SCD patients as they progress to VOC. Aim 2: Determine the effect of acupuncture on inflammatory/immune patterns. Aim 3: Investigate the anti-inflammatory mechanisms of acupuncture. Persistent disparities in healthcare access faced by African Americans are widely recognized and have clear links to unfavorable health outcomes. If successful, the proposed studies have the potential to unveil novel mechanistic targets for effectively managing VOC and advance our understanding of the mechanisms underlying acupuncture analgesia.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY / ABSTRACT Unstructured clinician notes contain “80% of medical information” in the electronic health record (EHR) but are cumbersome to search and analyze. Multiple national thought leaders in health care, including the National Academy of Science, Engineering and Medicine have called for improved “access, tools, and capacity for [clinical] data analysis.” Yet, our inability to search, summarize, and extract knowledge from clinical notes remains a major technical impediment to replace the tedious, time-consuming task of human chart review. Large language models (LLMS) are advanced deep learning algorithms capable of the understanding of concepts and context from text. They facilitate several tasks relevant to digital chart review such as advanced information search and retrieval. Furthermore, LLMs can generate coherent prose to form context specific summarizations. Together, these two abilities hold the potential of realizing the vision of a world where most clinical decisions are cost-effectively “supported by accurate, timely, and up-to-date clinical information.” Nevertheless, real world deployment of digital chart review must ensure performance (i.e. accuracy, reliability) and clinicians’ compatibility to facilitate effective and sustainable interactions with clinicians within the context of their clinical workflows. Our research goal is to develop a digital chart review for providing clinicians with accurate and actionable information at the point of care. Our central hypothesis is that a domain adapted LLMs can deliver a decision focused chart summary which is comparable to human chart review by vascular surgeons. We will test this hypothesis using real world data from a large statewide health information exchange on more than 124,000 patients with peripheral arterial disease i.e. PAD (the leading cause of amputation in the US). The mentorship panel developed a set of research activities to execute the following aims that are aligned with the candidate’s overall goal of becoming an independent clinical investigator capable of using advances in health information technology to transform vascular care, and thus the health outcomes of underserved communities living with PAD. In Aim 1 we design and validate a large Language Model capable of extracting and summarizing PAD information from plain text notes. We will evaluate the outputs of the large Language model for accuracy, completeness, relevance, and uncertainty using industry standard metrics. We hypothesize that the LLM text extraction and summarization is comparable to human chart review. In Aim 2, we Co-design with clinicians a minimally viable prototype of a digital chart review system. We will incorporate feedback from the clinicians using the iterative Agile Innovation process. We hypothesize that different clinical scenarios will require emphasis of different information types.

NIH Research Projects · FY 2025 · 2025-08

Abstract: Aging causes deterioration of the musculoskeletal system via disruptions in critical paracrine and endocrine systems. Sarcopenia, the loss of muscle mass and function, and osteoporosis, the weakening of the bone, predispose the elderly to debilitating injuries, such as fracture, disability, and other adverse outcomes associated with morbidity and mortality. The societal costs of sarcopenia, osteoporosis, and their associated complications will rise dramatically as the aging population rapidly increases in size. Since 2011, when the first baby boomers reached retirement age, the age 65-and-older demographic has markedly grown. According to the US Census Bureau, in 2034, for the first time, adults over 65 years of age will outnumber children under 18, creating an essential need for improved clinical care. While the elderly retain some ability for bone and muscle improvement with exercise intervention, the anabolic response of musculoskeletal tissues is attenuated in the aged. Thus, new therapeutic strategies are required since the disease mechanisms involving tissue crosstalk are not understood. The osteocyte is an endocrine cell that responds to systemic and local cues to provide critical regulation of muscle. We validated that contracted muscle secretes L-β-aminoisobutyric acid (L-BAIBA), which protected against bone and muscle loss during hindlimb unloading. Our model suggests that L-BAIBA is in- creased with muscle contraction during exercise, and propose that L-BAIBA acts on the osteocyte which in turn produces fibroblast growth factor (FGF) 23 to target the kidney in order to maintain normal phosphate homeo- stasis. L-BAIBA acts through the Mas-Related G Protein receptor type D (Mrgprd) to directly increase Fgf23. This receptor is almost completely down regulated during aging, suggesting that this event causes an altered muscle-osteocyte homeostatic axis, leading to altered phosphate via lowered FGF23. Our central hypothesis is: L-BAIBA is a critical mediator of osteocyte FGF23 induction via Mrgprd-mediated β-catenin signaling, and the effects of aging on this system cause phosphate driven pathologies that compromise muscle function. We expect our studies using dovetailed, cutting-edge in vivo and in vitro techniques to provide novel insight into the trans- lational biology of muscle and bone function during aging. Our studies also feature a critical role for the kidney in exercise-induced musculoskeletal adaptation via excretion and balance of excess phosphate generated during exercise or aging. If successful, our studies would support that L-BAIBA protects muscle via controlling osteocyte FGF23. These findings would suggest that interventions that target molecular regulation of phosphate balance, including inducing levels of L-BAIBA might have protective effects during aging onset, a syndrome of progressive Mrgprd down regulation, to increase FGF23 and thus improve muscle function and patient outcomes.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY Myocardial infarction (MI) causes 1 of every 7 deaths in the United States. Because the heart has little regen- erative capacity after injury, an urgent need exists for novel strategies in MI therapy. MicroRNAs (miRs) are small noncoding RNAs that downregulate targets, and miRs are being pursued as novel therapies for heart failure in clinical trials. Vascularization is crucial for cardiomyocyte (CM) survival after MI. Ischemic insult re- sulting from insufficient blood supply to the heart results in reduced myocardial capillary flow reserve and capil- lary density, impaired tissue perfusion, and augmented cellular injury and scar formation, leading to progres- sive left ventricular remodeling and chronic heart failure. Because CM renewal is absent in adult mammalian hearts, dead CMs after MI are also replaced by scar tissue. Although the significance of endothelial dysfunc- tion and CM dropout in MI has been established, identification of the effectors underlying endothelial cell (EC) injury, CM dysfunction, and EC–CM crosstalk remains incomplete. Our goal here is to elucidate the roles of the novel EC- and CM-specific axis in MI. Beta-arrestin-mediated beta2-adrenergic receptor signaling (beta2AR/beta-arr signaling) elicited myocardial protection and in part contributed to the beneficial effects of the nonselective beta-blocker carvedilol. We discovered that carvedilol, acting through this mechanism, pro- moted the maturation of endothelial-enriched miR-532-5p (miR-532). Circulating miR-532 is a potential bi- omarker for MI. A secreted serine protease called PRSS23 initiates endothelial-to-mesenchymal transition and is essential for the cardiac valve formation in zebrafish embryo. Using innovative mouse models, we have made key discoveries: (1) systemic loss of Prss23, a key effector of miR-532, almost completely protects from cardiac dysfunction post-MI; and (2) systemic, EC-expressed, or CM-expressed miR-532 contributes to beneficial remodeling in ischemic hearts, which is opposed by EC-derived or CM-derived Prss23. Thus, dis- secting the pleiotropic mechanisms of action of the miR-532/PRSS23 axis may lead to the development of novel effective therapies for MI. Our hypothesis is that miR-532 elicits autocrine and paracrine protective ef- fects on ECs and CMs against ischemic stress by inhibiting maladaptive PRSS23 and cell extrinsic signals. To test our hypothesis, we plan to pursue three aims: (1) test if PRSS23 is a critical target of endothelial miR-532 and if pharmacological inhibition of PRSS23 promotes neovascularization post-MI; (2) determine the functional requirement of the CM-derived miR-532/PRSS23 axis during both murine post-MI remodeling and human CM damage; and (3) elucidate the key paracrine mechanisms by which the miR-532/PRSS23 axis in ECs regu- lates CM survival after ischemia. This proposal is innovative because the proposed functional axis has never been studied in a cell-type specific manner or with genetically modified mice. This project also employs trans- lational human cell and pharmacological studies and is significant because the identified beta2AR/beta- arr/miR-532-mediated PRSS23 regulatory mechanisms can be exploited as a novel target for MI.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Lymphedema is chronic limb swelling from lymphatic dysfunction which affects 250 million people worldwide. It is estimated that 5-10 million Americans have lymphedema. Primary lymphedema is rare and results from errors in lymphatic development. Secondary lymphedema accounts for 99% of the condition and occurs most commonly following surgical management of solid tumors (e.g., breast cancer, melanoma). Axillary lymph node dissection results in lymphedema in 30% of patients post-operatively. Skin thickening, interstitial fluid retention, and fibroadipose subcutaneous deposition from inflammation result in progressive limb enlargement. Lymphedema impacts quality of life and has a high health cost burden. Morbidity includes recurrent cellulitis, pain, and impaired extremity function. Non-surgical management of lymphedema includes compression therapy. Surgical treatment involves excisional (e.g, skin/subcutaneous resection) and microsurgical physiologic procedures including vascularized lymph node transfer and lymphovenous bypass. Current treatments may variably improve limb size, but do not cure lymphedema. Lymphatic capillaries develop discontinuous button-like junctional proteins which facilitate absorption of interstitial fluid into lymphatics at the capillary level. Continuous zipper-like junctional proteins are present in lymphatics which transport lymph without leakage. Rare genetic causes of primary lymphedema affect initial lymphatic capillary button junction development resulting in zippers instead of buttons. In the lung, chronic pleural inflammation has been found to result in the replacement of button junctions in pleural lymphatics with zipper junctions leading to impaired drainage function and pleural effusion. Secondary lymphedema occurs on an average of one year post-operatively following surgical lymph node dissection indicating the etiology of lymphedema is not only the mechanical disruption of lymphatics. Following post-surgical lymphatic injury, activation and maintenance of chronic inflammation is critical in the development of lymphedema. The goal of this application is to understand the mechanism and pathophysiology of junctional protein changes in secondary lymphedema. This will inform us about the fundamental underlying etiology of lymphatic stasis in lymphedema and, importantly, help us develop targeted therapies to improve the lives of patients with lymphedema. We hypothesize that inflammation after lymphatic injury results in capillary lymphatic button junction changes to impermeable zipper junctions rendering interstitial fluid uptake inefficient. In Aim 1, we elucidate how lymphatic injury affects lymphatic junctional proteins in secondary lymphedema. In Aim 2, we determine the effects gene delivery targeting junctional protein pathways following lymphatic injury. Our approach uses a combination of mouse models and clinical specimen to have a high translational significance.

NIH Research Projects · FY 2025 · 2025-08

Monoclonal antibodies (mAbs) targeting infectious agents act primarily by direct neutralization but their ability to bind to specific Fc receptors can either extend antibody half-life or enhance cellular activity against the pathogen. The anti-malarial IgG1 mAbs CIS43LS and L9LS are effective at preventing malaria infection in randomized, placebo-controlled clinical trials conducted in individuals who are naturally exposed to Plasmodium falciparum malaria. Both mAbs have increased binding affinity to the neonatal Fc receptor (FcRn), resulting in enhanced IgG recycling and thus longer serum half-life, while retaining the ability to bind to Fcγ receptors (FcγRs). Significant variability in anti-malarial mAb half-lives has been observed in clinical trials, which has implications for the optimizing dosing to extend duration of efficacy. We hypothesize that specific innate signals alter the protective efficacy of anti-malarial mAbs by regulating Fcγ-mediated effector responses or by extending half-life via enhanced FcRn-mediated recycling. To address this, we will leverage prospective clinical data, pharmacokinetic data, and biospecimens from a phase 2 pediatric trial of L9LS to conduct an unbiased, comprehensive analysis of transcriptomic, metabolomic, and cytokine profiles in blood obtained from trial participants before and after receipt of L9LS or placebo. Differential analysis and integrated, multi-omics machine learning will be conducted to determine immune features predictive of L9LS-mediated malaria protection and longer mAb half-life. Immune cells from malaria-protected and malaria-susceptible individuals will be compared using 1) single-cell transcriptomics to assess for differentially activated states within specific cell types and 2) in vitro functional assays to assess antibody-dependent cellular phagocytosis and cytotoxicity. Specific findings will be validated using samples from independent L9LS and CIS43LS trials conducted in malaria-exposed and malaria-naïve adults. The study has the potential to identify molecular signatures predictive of mAb-mediated protection from P. falciparum infection and address fundamental questions applicable to other anti-infective mAbs. Findings from this proposal could determine the impact of innate immune activation, which can vary even in states of apparent health, on the efficacy of anti-infective mAbs and provide insight on potential Fc-mediated mechanisms of action. Determining host features that affect the pharmacokinetics of the mAbs L9LS and CIS43LS has broader implications for understanding the dosing, clearance, and efficacy of other IgG1 mAbs. Successful completion of this project may also reveal strategies for enhancing and prolonging the therapeutic efficacy of IgG1 mAbs targeting other pathogens and clinical conditions. Individualizing mAb dosing based on patient-specific molecular characteristics can potentially reduce costs and improve resource utilization, which may be critical during outbreaks of emerging pathogens or in resource-limiting settings.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Hearing loss (HL) is the most common sensory disorder worldwide, with significant genetic, allelic, and phenotypic heterogeneity. Due to its complexity, HL presents many challenges in diagnosis, management, and treatment, with over 220 genes and 16,000 variants implicated in its nonsyndromic and common syndromic forms. The Hearing Loss Clinical Domain Working Group (HL-CDWG) established within the Clinical Genome Resource (ClinGen) has addressed these challenges through the curation of gene-disease relationships (GDRs) and assessment of the pathogenicity of variants associated with HL to enhance the diagnostic accuracy and clinical care of persons with HL. This work is particularly timely, given the rapidly increasing number of genes identified for clinical testing and ongoing clinical trials for genetic HL. The HL-CDWG is an international, multidisciplinary team of over 50 members from 40 institutions across six continents, specializing in the key areas of expertise with representation of research, clinical practice, and diagnostic laboratories. This group consists of HL experts - geneticists, basic research scientists, auditory biologists, otolaryngologists, audiologists, and genetic counselors - along with gene curation framework experts and biocurators ensuring a comprehensive approach to addressing genetic HL. The HL-CDWG is structured into the HL Gene Curation Expert Panel (GCEP) and the HL Variant Curation Expert Panel (VCEP). These panels have collaborated to refine the evidence supporting GDRs and variant classification. In this proposal, we will build upon and expand the activities of the HL-CDWG. In Aim 1, we will thoroughly curate the evidence supporting GDRs and quantify their strength using the ClinGen framework. In Aim 2, we will develop standardized, evidence-based gene-specific variant classification guidelines specifically tailored to each gene's mutational signature and molecular mechanism. We will prioritize genes targeted in gene therapy-based clinical trials and those with pleiotropic effects. This effort enhances the precision of genetic testing and interventions, aligns with successful clinical trials for genetic HL, and marks a pivotal step towards precision medicine. The work of the Hl-GCEP and VCEP will be disseminated through publications, webinars, and workshops at meetings targeted at the clinical and research communities in otolaryngology and the broader genetic community. This strategy ensures the effective implementation of an evidence-based framework and practices to enhance patient care and improve health-related outcomes and aligns perfectly with the 2023-2027 strategic plan of the National Institute on Deafness and Other Communication Disorders.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT The candidate’s long-term goal is to shift his research focus from examining neural correlates of individual differences in adolescent substance use disorders toward neurobiological predictors of individual treatment outcomes. Through the research and training in this K23 proposal, the PI will acquire training in multivariate statistics/machine learning methods and clinical research necessary to make this transition. Adolescent cannabis use is a serious public health problem. 6% of high school seniors report daily cannabis use and 3% of adolescents meet criteria for CUD. Although cannabis is increasingly becoming legal at the state level, cannabis is associated with increased risk of psychiatric disorders, cardiovascular disease, and respiratory disease. However, CUD treatment responses are inconsistent. Contingency management (CM) is a critical component of CUD treatment and relies on instrumental learning to reduce cannabis use by rewarding abstinence from cannabis use. However, little work has investigated whether integrity of brain networks underlying instrumental learning predict CUD symptom severity, and no work has investigated whether performance on an instrumental learning task predicts cannabis use outcomes following CM. Therefore, the proposed K23 project will address the candidate’s training goals while gathering preliminary and feasibility data supporting the next step in his research. The proposed research study aims to: (1) identify associations between CUD symptomatology and neural activity during instrumental learning and (2) examine whether neural activity during instrumental learning is associated with reduced cannabis use frequency during CM treatment. The PI proposes to accomplish these aims by collecting fMRI and report-based data from a sample of 76 youth ages 14-18 who use cannabis at least once per week. All youths will complete a screening visit, an fMRI scanning visit, 10 virtual CM sessions, and a follow-up visit. We anticipate that findings will show the extent to which disruptions in neural systems underlying instrumental learning are associated with CUD symptoms and the extent to which neural activity instrumental learning predicts contingency CM outcomes. This K23 application proposes training and research that is directly in line with NIDA objectives and represents a logical progression from the PI’s prior experience to address career development goals in three areas: (1) use of multivariate and machine learning techniques in analyzing neuroimaging data; (2) clinical research. The experience and findings from the proposed K23 will position the PI to pursue NIH funding and build this line of research through a large mechanistic clinical trial of CM treatment on adolescent cannabis use and the role of neuroimaging data in individualized treatment outcomes.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Enteric viruses are infectious human pathogens that lead to significant disease and symptoms such as nausea, diarrhea, vomiting, and abdominal pain. These viruses spread through the fecal-oral route, initiating infection in the intestine and traverse through the gastrointestinal tract. Enteric viruses are easily spread by contamination of food and water and by contact between individuals. Coxsackievirus is a type of enteric virus that causes hand, foot, and mouth disease, aseptic meningitis, conjunctivitis, and myocarditis. Specifically, Coxsackievirus B3 (CVB3) is the major isolate associated with viral myocarditis. Because CVB3 is an enteric virus and initiates infection in the gastrointestinal tract, the virus encounters a diverse population of intestinal bacteria. Previous studies have shown that CVB3 interacts with bacteria and that this interaction enhances viral stability, infectivity, replication, and pathogenesis. Our lab has also shown that bacteria-mediated enhancement is specific to bacterial species as well as to species-specific bacterial cell wall components. Notably, Salmonella enterica was able to enhance CVB3 infectivity, but not Escherichia coli. Furthermore, specific bacterial cell wall components are able to enhance CVB3 stability. Smooth type LPS from S. enterica was able to enhance CVB3 stability, but not rough type LPS nor any LPS from E. coli strains. Interestingly, CVB3 is able to bind to both S. enterica and E. coli despite these differences in bacteria-mediated viral enhancement. Previous data demonstrate that enteric viruses can bind directly to bacteria, including CVB3. However, it is unknown if binding trends affect bacteria-mediated viral enhancement or if specific components of bacterial cell walls are responsible for binding. The overarching goal of this project is to characterize the binding interaction between CVB3 and enteric bacteria. Our central hypothesis is that CVB3 binds enteric bacteria through specific LPS structures that dictate binding affinity. To examine this hypothesis, I will perform biochemical binding assays to identify the specific bacterial cell wall structures that allow for bacterial binding to CVB3. I will also compare binding affinities between CVB3 and different intestinal bacteria. Experiments outlined in Aim 1 will establish the role that LPS plays in binding to CVB3 and identify the specific components of LPS that are required for CVB3 binding. Experiments described in Aim 2 will widen the scope of this study by examining differences in CVB3 binding trends to different intestinal bacteria and LPS structures from different bacterial species. Overall, these data will distinctly characterize the interactions between CVB3 and enteric bacteria. Completion of this project will reveal more about the mechanisms driving bacteria-mediated enhancement of CVB3 and will provide insight into potential conserved features between CVB3 and other enteric viruses. This will increase our understanding of the pathogenesis of enteric viruses and guide future therapeutic avenues.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Chronic kidney disease (CKD) affects nearly 1 in 7 people and leads to a 5-fold increased risk of fracture as compared with normal individuals. Disease progression leads to changes in bone and mineral metabolism, including prolonged elevations in parathyroid hormone (PTH), resulting in bone resorption. Despite clinical efforts to control PTH-induced bone loss, fracture risk in CKD patients has not changed. This indicates additional mechanisms are at play. We hypothesize a role may be played by myokines-- compounds produced by muscle-- that act on bone, including irisin. Irisin has been shown in healthy animal models to upregulate bone formation transcription factors and improve cortical bone mass. Our preliminary data suggests that the high PTH in CKD-MBD inhibits the positive effects of irisin and/or its precursor (Fndc5) on bone. We will test our hypothesis by 1) investigating the mechanism of this relationship in vivo in a CKD-mineral bone disorder (MBD) rat model and in vitro using cultured osteocytes and 2) validating this relationship in humans using muscle and bone biopsies from patients with CKD. This interplay may be a missing link to explain alterations in bone remodeling, offering a new therapeutic strategy for patients with CKD-MBD.

NIH Research Projects · FY 2025 · 2025-08

Project Summary This proposal identifies the EphB2 receptor tyrosine kinase and its cognate EphrinB ligands as potential therapeutic targets to prevent vascular damage and fibrosis associated with systemic sclerosis (SSc, also known as scleroderma). A hallmark of SSc is the progressive and overwhelming deposition of extracellular matrix components, especially collagen, to cause the skin to become fibrotic and lose its elasticity. This process is thought to be driven by the recruitment of immune cells to sites of tissue damage, providing an inflammatory microenvironment to enhance fibroblast-to-myofibroblast transitions that in SSc patients leads to pathological expansion of pro-fibrotic cells and massive upregulated expression of collagen and other genes involved in fibrosis. As the biochemical pathways that control these events remain incompletely described, we focused our attention on potential membrane-associated molecules that may help interpret extracellular signals and aid the conversion of dermal fibroblasts into fibrogenic myofibroblasts, and identified the EphB2 receptor interacting with its EphrinB ligands as possible important components. Emerging data support the involvement of EphB-EphrinB in fibrosis of multiple organs, including our previous work, however little is known about the potential role of these highly conserved signaling molecules in the pathogenesis of SSc. Using human skin biopsies and mouse models of skin fibrosis, we will test the hypothesis that upon chronic, immune-mediated skin injury, EphB2 expression becomes strongly upregulated and the enhanced signaling pathways activated by this molecule are critical for the transdifferentiation of quiescent dermal fibroblasts into fibrogenic myofibroblasts to help bring about skin fibrosis. Our general idea is that when bound to EphrinB ligands expressed on various cells of the injured skin microenvironment (including endothelial cells), activated EphB2- expressing fibroblasts will initiate a differentiation process leading to their transformation into pro-fibrotic myofibroblasts. In support of this, preliminary data is provided that shows EphB2 expression is highly upregulated in human skin from SSc patients and in normal human dermal fibroblasts exposed to the pro- fibrotic inflammatory cytokine TGF-β1, and that skin fibrosis can be modulated by disrupting EphB2 either through genetic mutation or novel pharmacological approaches. The preliminary data has guided the formulation of three Specific Aims that will further test our ideas. Aim 1 will determine whether activation of EphB2 forward signalling is required for the progression of skin fibrogenesis. Aim 2 will test the hypothesis that EphB2-EphrinB interactions and signaling contributes to vascular damage and defective angiogenesis in SSc. Finally, Aim 3 will determine whether therapeutic targeting of these molecules will mitigate skin fibrosis. The proposed research is highly significant and innovative as it will reveal a key molecular mechanism that drives excessive fibrosis in SSc, and will also provide important early data on how novel small molecules that target EphB2-EphrinB interactions and signaling could be employed as future therapies to treat SSc.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Individuals with Trisomy 21 (Ts21) have ~80 defined clinical phenotypes collectively known as Down syndrome (DS); the incidence and severity of these phenotypes, however, are highly variable. The relationship between specific genes and phenotypes associated with DS has been studied for decades, but how modifier genes affect the incidence and variability of DS phenotypes is not well known, and only a few attempts have been made to identify two-copy or background genes that significantly modify DS phenotypes. Additionally, differences in many skeletal and neurobehavioral phenotypes have been noted between males and females with Ts21, but how sex modifies DS phenotypes has been understudied. More than twenty DS mouse models have been developed to understand gene-phenotype relationships in DS, but the inbred genetic backgrounds of all current mouse models do not offer an adequate landscape to analyze the impact of genetic diversity seen in humans with DS. Moreover, limited study of the development of sex-specific phenotypes has hampered the identification of the modifying causes of these traits. Additionally, how changes in the skeletal system may affect cognitive and behavioral functions has not been extensively investigated. Our central hypotheses are that novel, genetically robust DS mouse models will better represent the human condition and will facilitate localization of modifier genes and molecular mechanisms linked to the variable sex-specific skeletal and cognitive phenotypes associated with DS. The objectives of this project are to create new DS mouse models that better represent the variability found in the human Ts21 genetic condition, identify modifier genes and molecular pathways that lead to the variability in incidence and severity of DS skeletal and neurobehavioral traits, and determine if skeletal improvements ameliorate brain-related phenotypes in DS mice. To achieve these objectives, we will 1) create a diverse set of new DS mouse model lines to determine the overall contribution of triplicated and non-triplicated background genes on the incidence and severity of skeletal phenotypes associated with DS; 2) utilize an innovative QTL analysis of the new DS mouse model lines to identify non-triplicated modifier genes most affecting the incidence and severity of sex-specific DS skeletal phenotypes; 3) quantify modifiers of and changes skeletal and neurobehavioral measures after mechanical stimulation of bone in a selection of the new DS mouse model lines. Our proposal is significant because it establishes methodology to define gene-phenotype relationships for any DS phenotype, quantifies mechanisms of modifying gens, and determines if there is a link between abnormal DS skeletal development and functional brain deficits. This innovative work is in response to the NIH INCLUDE (INvestigation of Co- occurring conditions across the Lifespan to Understand Down syndromE) request for new DS mouse models to better define the pathophysiological effects of Ts21 and proposes a high-risk basic science study of the co- occurring bone and brain traits to identify genetic pathways that may be most responsive to new DS therapies.

NIH Research Projects · FY 2025 · 2025-08

A radiological or nuclear incident (e.g., an accident at a nuclear reactor site or a terrorist attack involving the detonation of an improvised nuclear device) could result in the exposure of thousands of people to lethal or potentially lethal doses of ionizing radiation (IR). While good progress has been made in developing mitigators to treat individuals that received doses high enough to trigger the hematopoietic acute radiation syndrome (H- ARS), there are no approved strategies to mitigate the delayed effects of acute radiation exposure (DEARE) that would subsequently manifest in H-ARS survivors. Several life-threatening or debilitating cardiac DEARE- related pathologies are observed months to years after the heart is exposed to doses known to elicit symptoms of the H-ARS. Thus, it is necessary to develop compounds or strategies that can mitigate cardiovascular dysfunction if administered 24 h or more after irradiation, involve easy deployment and use protocols, and are non-toxic. We previously identified and partially characterized a novel non-pharmacological strategy which we found to be very effective at mitigating the lethal effects of total body irradiation (TBI) in mice if administered after exposure. Our approach involves the creation of a small (3 mm) subcutaneous (SC) incision post- irradiation. We found that SC wounding several minutes after exposure to a range of IR doses greatly protected against lethality, and that mitigation of IR effects and the resulting H-ARS may be mediated by enhanced recovery of hematopoiesis (likely due to modulation of specific cytokines). We refer to this strategy as “protective wounding.” Most recently, we accumulated preliminary data which also suggest that cardiac function measured 30 days post-irradiation is preserved in mice that receive a SC cut after irradiation, compared to mice that do not. We propose to use “protective wounding” as a novel model for interrogating the proteins modulated and pathways involved in the mitigation of cardiotoxicity induced after TBI exposure. We will first determine optimal parameters after irradiation for our wounding model that will produce the greatest mitigation of cardiovascular dysfunction, and demonstrate the validity and applicability of the strategy as a mitigator of IR-induced cardiotoxicity. To accomplish this, various endpoints of cardiac function/structure in mice that receive a cut up to 5 days after receiving a single TBI exposure will be compared with sham-cut mice. We will next determine whether SC wounding (after exposure to a TBI dose expected to result in cardiotoxicity in H-ARS survivors) results in changes in cytokine and cardiac proteomics profiles that impact post-irradiation cardiovascular damage or recovery by correlating cytokine and proteomics data with structural and fuctional cardiovascular endpoints. Data accumulated will then be used to identify the pathways and begin to identify the mechanisms by which IR-induced effects on cardiac structure and function may be mitigated. These experiments should ultimately lead to the development of pharmacological mitigators of IR-induced cardiotoxicity that can be administered to mass casualties 24 h or more after a radiological/nuclear incident.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT CANDIDATE: Dr. James's career objective is to establish an independent research group investigating mitochondrial dysfunction (MD) and Aminopeptidase N (APN) driven signaling in pulmonary hypertension (PH). The overall research and career development plan is carefully crafted to impart the necessary skills for Dr. James to establish a novel and independent line of research. The plan includes the following objectives: 1) gain additional training in research methodology and professional development; 2) developing strong publication records; 3) presenting his work at scientific meetings to make an impact on the field 4) obtaining preliminary data for his first independent grant. ENVIRONMENT: The primary mentor Dr. Rafikov is an expert in mitochondrial dysfunction and PH. He will provide guidance and technical expertise for aspects in all aims. Co-Mentor Dr. Vinicio de Jesus Perez is an expert in pericyte biology and PH and will train Dr. James in pericyte isolation, co- culture, and data analysis. Co-mentor Dr. Ralph Fregosi is an expert electrophysiologist and will extensively prepare Dr. James in his career development in addition to training with patch clamp techniques. All the mentors with the rest of the advisory committee will ensure the success of Dr. James's proposal. All the proposed work will be carried out in Dr. Rafikov's laboratory with training gained from labs of Dr. de Jesus Perez and Dr. Zhiyu Dai for single cell transcriptomics. Dr. Rafikov has all the necessary funding from NIH and other sources to support Dr. James during the K99 training phase. The Department of Medicine at the University of Arizona has excellent facilities and supportive faculty members and provides a nurturing environment for the training phase. RESEARCH: The overarching hypothesis is that MD triggers the overexpression of the protein, APN in pericytes. APN has been shown to cause migration, invasion, and proliferation of cells. Preliminary data suggests that MD in rats triggers an increase in pericyte APN, causing them to dissociate from endothelial cells (EC's) in capillaries, transform into smooth muscle-like cells (PASMC) and proliferate. This ultimately leads to vascular remodeling and PH. Three specific aims will address this hypothesis. Aim 1: To Investigate the role of APN in pericyte-EC communication and pericyte transformation to PASMC. Aim 2: To Investigate the role of MD on APN expression and signaling. Aim 3: To Verify the impact of blocking APN and reversing MD on vascular remodeling in PH. The study will utilize several techniques including single cell transcriptomics, 3D imaging and co-culture to address these questions. The results of this study will lead to a better understanding of how MD drives APN expression and signaling. This could define novel targets in the treatment of PH for favorable clinical outcomes. The proposed training will enable Dr. James to acquire new experimental skills and execute his research plan successfully. This will allow Dr. James to become self-sufficient, able to secure extramural funding and a tenure- track position at a top-tier research institution.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT The goal of the proposed five-year training plan is the development of my independent research career as an academic hematopathology physician-scientist studying hematopoietic stem and progenitor cell (HSPC) crosstalk and transplantation. I am currently a research fellow at the Children’s Hospital of Philadelphia (CHOP) and attending physician in the Division of Hematopathology at the University of Pennsylvania (UPenn). I am specifically seeking to develop and refine skills required for my successful career as an independent investigator, including expertise in HSPC transplantation and microscopy, miRNA mimic and inhibitor screening, lipid nanoparticle delivery, and bioinformatics. My overarching goal is to improve HSPC transplantation outcomes through study of key mechanisms of HSPC-niche cell crosstalk. My mentor for this award is Dr. Peter Kurre, a recognized leader in hematopoietic stem cell biology and bone marrow (BM) failure syndromes. To further add to my scientific and career guidance, I have assembled a mentoring committee composed of scientists and physician-scientists from complementary research fields. I will have the full resources and support of UPenn and CHOP available for the completion of my research and career development goals. This proposal outlines experimental plans to define mechanisms of cellular crosstalk between HSPCs and BM ECs that will inform potential strategies for improving HSPC transplantation. Graft success after transplantation requires initial homing of donor HSPCs to the bone marrow niche, a specialized vascular microenvironment that supports long-term blood cell and immune system reconstitution. BM migration is a dynamic process involving various adhesion and signaling molecules that are not completely defined. My preliminary data have elucidated a novel mechanism of crosstalk between HSPCs and ECs that initiates from cargo trafficked by secreted extracellular vesicles (EVs). Specifically, HSPC EVs promote remodeling of ECs and lead to reciprocal CCR2 receptor-ligand interactions that likely interact with CXCR4 to drive HSPC migration. This proposal will define the molecular mechanisms underlying EC remodeling and HSPC migration, and investigate innovative pre- conditioning strategies to recapitulate and exploit physiologic mechanisms with the goal of improving HSPC transplantation outcomes. These objectives will be achieved via three specific aims: to elucidate key signaling pathways driving EC remodeling, to define CCR2 cooperativity in BM homing, and to test pre-conditioning strategies to induce EC CCR2 ligand upregulation and enhance HSPC engraftment. The outcome of these studies will deepen our understanding of HSPC-niche crosstalk and unveil new therapeutic strategies to optimize HSPC transplantation. Completion of these aims will consolidate my experience in models of murine HSPC transplantation and EV-mediated signal transduction, further my training in nanoparticle-based in vivo delivery, and provide the basis for a future R01 funding proposal, leaving me uniquely prepared for an independent career as a physician-scientist with a focus on HSPC transplantation.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Neurofibromatosis type 1 (NF1) is a cancer predisposition syndrome caused by a mutation in the NF1 tumor suppressor gene that occurs in 1:3000 live births. Loss of neurofibromin, a GTPase activating protein for p21 RAS, causes hyperactivation of the RAS pathway that leads to the formation of plexiform neurofibromas (PNF) in up to 50% of individuals with NF1. In the majority of NF1 patients, PNF grow slowly, typically reaching stability in adulthood. However, a subpopulation of NF1 patients experience progression to atypical neurofibromatous neoplasm of uncertain biologic potential (ANNUBP) and transform to malignant peripheral nerve sheath tumor (MPNST). MPNSTs are the leading cause of premature death in NF1 patients with a 5-year survival rate of 20- 50%. Therefore, understanding the molecular and cellular interactions that mediate progression along the peripheral nerve sheath tumor (PNST) continuum is critical to improving NF1 patient survival. The genetic events governing neurofibroma progression and malignant transformation have been extensively studied and several key driver events have been implicated. However, preclinical and natural history studies suggest that these genetic changes may not fully account for the clinical heterogeneity of PNF and ANNUBP precursor lesions and their evolution to MPNST. Therefore, I postulate that additional factors either intrinsic to neoplastic Schwann cell progenitors or within the tumor microenvironment play a critical role in modulating the growth and malignant potential of neurofibroma. My recent work defining the spatial gene expression profile of human NF1-associated tumors identified alterations in signatures of immune surveillance, including T cell activation and cytotoxicity, across the PNST continuum. Anti-tumor T cell response signatures were upregulated in ANNUBP, whereas precursor lesions that progressed to MPNST were characterized by signatures of T cell exhaustion. Furthermore, immunofluorescence staining showed that human ANNUBPs were characterized by increased infiltration of CD4+FOXP3- and CD8+FOXP3- T cells, while FOXP3+ T cells (T regulatory cells) predominated in MPNST. While the density and functional states of T cells infiltrating the tumor microenvironment are important predictors of therapeutic response and prognosis in many human cancers, the functional role of distinct T cell subsets in governing neurofibroma progression to MPNST remains undefined. The objective of this work is to define the T cell subsets present at various stages of the PNST continuum and investigate their functional role in tumor progression. I hypothesize that effector CD4 and/or cytotoxic CD8+ T cells are critical in preventing malignant transformation of PNF and ANNUBP precursors. Ultimately, these studies will advance our understanding of the functional role of T cell subsets across the PNST continuum and inform future diagnostic and therapeutic strategies for patients with NF1.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Pediatric asthma is a growing epidemic associated with significant burden to the healthcare system which lacks individualized treatment approaches. Although standard treatments for pediatric status asthmaticus are adequate for most patients, a subset do not respond quickly. These children are at risk for severe physiologic derangements including hypercarbia, hypoxemia, cardiac dysfunction, and death. Adjunctive asthma therapies can potentially provide benefit to children with status asthmaticus; however, there is limited evidence regarding their efficacy. Not only are these adjunctive therapies potentially ineffective, but many are associated with severe side effects. My central hypothesis is that children presenting to the hospital with status asthmaticus can be effectively subdivided into distinct clinical phenotypes that exhibit heterogeneous treatment effects. I propose to test this hypothesis by (1) identifying distinct clinical phenotypes in children presenting to the hospital with status asthmaticus, (2) validating the clinical phenotypes of status asthmaticus in other pediatric institutions, and (3) comparing effects of the adjunctive asthma treatments magnesium and aminophylline between clinical phenotypes. With the preliminary data generated, new skills obtained, and expertise gained through this K01 award, I plan to construct a robust computational phenotype for pediatric asthma, identify the patients most likely to have improved clinical outcomes with specific asthma therapeutics, and decrease the health burden of children with status asthmaticus in a future R01 study. During the award period, I will continue to conduct research under the primary mentorship of Dr. Eneida Mendonca, who is a pediatric critical care physician and established health informatics researcher, as well as the co-mentors and advisors outlined in the career development plan. I will obtain additional training in artificial intelligence and machine learning methods such as unsupervised clustering methods, health informatics and information systems, and comparative effectiveness research methods through both formal coursework as well as established institutional programs. With the preliminary phenotyping data generated by this study, the experience gained using health informatics and machine learning methods, and a knowledge foundation of comparative effectiveness research methods, I will be well-positioned to transition to research-independence.

NIH Research Projects · FY 2025 · 2025-07

An individual’s phenotypes related to their health conditions are associated with the complex interplay between the individual’s genetics and their exposures to both internal and external environments. Nevertheless, genetics only account for ~10% of an individual’s health, while the remaining appears to be determined by environmental factors and gene-environment interactions. To comprehensively understand the causes of diseases and prevent them, environmental exposures, especially the spatial and contextual exposome—the contexts in which the person lives their life or external agents to which the one is exposed (e.g., environmental pollutants), need to be systematically explored. Nevertheless, the heterogeneous definitions of the spatial and contextual exposome and the heterogeneity of their data sources require us to adopt semantic standards using an ontology-driven approach to (1) provide an unambiguous and consistent understanding of the variables in heterogeneous data sources, and (2) explicitly express and model the context of the variables and relationships between them. On the other hand, the rapid adoption of electronic health record (EHR) systems has made large collections of real-world data (RWD) that reflect the characteristics and outcomes of the patients being treated in real-world settings, available for research. The increasing availability of RWD combined with the advancements in analytical methods, especially artificial intelligence (AI) and machine learning (ML) offer unique opportunities to generate real-world evidence (RWE). There is also an increasing interest in spatiotemporally linking spatial and contextual exposome data to real-world observational data including RWD to answer various questions on how exposures to environmental factors affect health status, disease development, and outcomes. However, there are key gaps in the research infrastructure to support these studies. Our long-term goal is to develop and disseminate methods and tools to advance spatial and contextual exposome research with RWD. Responding to RFA-ES-23-002, the objective of this proposal is to develop an innovative SPATIAL AND CONTEXTUAL EXPOSOME SEMANTIC DATA INTEGRATION SYSTEM (SPACESCANS) with (1) an ontology-annotated knowledge graph of existing publicly available high-quality spatial and contextual exposome data from heterogeneous sources, along with (2) a user-friendly data integration tool that can guide the users to choose (i) the appropriate spatial and contextual exposome variables, and (ii) appropriate spatiotemporal linkage methods to link exposome data with their RWD, based on their study needs; and (3) generate analysis-ready (and AI/ML-ready) RWD-exposome linked datasets for downstream analyses.

NIH Research Projects · FY 2025 · 2025-07

Project Abstract We request funds to upgrade our current Siemens MAGNETOM Prisma 3T MRI scanner (installed in 2014) to the Siemens MAGNETOM Cima.X Fit 3T MRI scanner. The Cima.X Fit scanner is a commercialized connectome scanner, designed for advanced neuroimaging, with a gradient system of 200 mT/m, whose prototypes were developed by Siemens Healthineers in collaboration with Massachusetts General Hospital, Cardiff University, and the Max Planck Institute for the human connectome project (HCP). Our request for an MRI system upgrade is driven by the need for advanced diffusion imaging techniques to probe brain and other tissue microstructures with greater detail and specificity and the need to leverage AI-assisted fast imaging. The gradient strength limitations of our current Prisma scanner prevent us from pursuing cutting-edge diffusion microstructural imaging. Over 90% of our projects utilize diffusion-weighted MRI. Building on our strong and well-funded neuroimaging program, the high-performance gradient system will elevate our research capabilities by enabling the bench-to-bedside translation of high-resolution and advanced diffusion MRI technologies that are unavailable to us now. The high-performance gradient system will allow us to move beyond conventional diffusion tensor imaging (DTI) with non-specific metrics and apply biologically specific metrics (e.g., soma fraction, neurite density, and axonal diameter) to our large clinical populations. This will enhance our translational science by further bridging the gap between basic neuroscience research on our preclinical small animal 9.4T MRI scanner and human imaging studies. In addition, the current MRI pulse sequence software has become obsolete, preventing us from applying advanced fast imaging technologies using compressed sensing and AI-assisted reconstruction. Fast imaging improves all our imaging studies (including neuro and body imaging), and shorter scan times, while maintaining image quality, will markedly reduce the participant burden. This is particularly important for neonates, older adults, and other clinical populations who may have difficulty tolerating longer scans.

NIH Research Projects · FY 2025 · 2025-07

PROJECT ABSTRACT/SUMMARY IU MAP-AD Overall Abstract/Summary The Indiana University Microphysiological systems to Advance Precision medicine for AD/ADRD (IUMAP-AD) Center is committed to establishing a multi-component Alzheimer’s disease and Alzheimer’s disease related dementias (AD/ADRD) Translational Center for Microphysiological Systems (MPS). AD is a leading cause of dementia worldwide, with current estimates of nearly 6 million Americans currently affected by the disease. AD is associated with many accompanying pathologies including inflammation and activation of microglia and astrocytes, dysfunction of vasculature, as well as synaptic and eventual neuronal loss. Additionally, recent studies have identified numerous genetic risk factors associated with AD/ADRD at loci with known functions in glial and vascular cell types. However, our understanding of how these genetic risk factors and cell types contribute to AD/ADRD pathology remains somewhat limited, especially in the human system. Thus, to overcome these shortcomings, we propose to build on our extensive knowledge and tools acquired from the ongoing MODEL-AD, TREAT-AD, and CLEAR-AD consortia at Indiana University, as well as our extensive expertise in induced pluripotent stem cell (iPSC)/organoids, intelligent systems, and AD/ADRD research across the University. Our overarching goal is to develop reproducible, scalable, and standardized 3D brain organoid-based MPS models that recapitulate key features of human AD/ADRD pathophysiology. In the process, we will also generate numerous resources for investigation among the research community which will be readily shared, including novel cell lines, protocols for the development and implementation of MPS models, and datasets derived from the use of MPS models. These models will thus serve as precision medicine research tools to investigate the complex biology of AD/ADRD and accelerate multiple aspects of drug discovery and preclinical drug development.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Oral inflammatory conditions such as Chronic graft versus host disease (cGVHD) and Oral Lichenoid Lesions (OLL) commonly affect oral exocrine tissues including minor salivary glands (MSG) that require incisional tissue biopsy for diagnosis. Saliva, a biofluid with complex composition, is secreted by the acinar cells of salivary glands to help in important oral functions such as lubrication, digestion, and importantly, to elicit anti-inflammatory responses providing the potential to present specific biomarkers that reflect health and disease status. Being able to collect saliva through patient-friendly, non-invasive methods is an important advantage of utilizing this biofluid for oral and systemic disease diagnostics. Recent shotgun proteomic screening work from our laboratory identified a significant reduction in protein content of zymogen granule 16B (ZG16B), a jacalin-related lectin protein, at onset of salivary gland cGVHD compared to healthy donors. Lectin proteins have a carbohydrate domain and adjacent positively charged binding pocket that can facilitate cell-to-cell interactions and immunomodulation contributing to important oral cavity tissue homeostasis. Therefore, we hypothesize that ZG16B is required to coordinate the localization and release of acinar cell secretions. Changes in ZG16B levels in states of altered oral tissue homeostasis can signal salivary gland damage. Aims 1 and 2 focuses on determining the distinct binding partners of ZG16B in human salivary gland (HSG) cells and conditioned media as well as its functional consequence. This will be fulfilled by endogenous co-immunoprecipitation, mass spectrometry, loss-of-function and gain-of-function experiments followed by entire RNA sequencing analysis. Aim 3 will quantify the changes in concentration of salivary ZG16B using sex-matched samples from healthy individuals and saliva from patients with various oral inflammatory conditions damaging salivary gland (oral cGVHD and OLL). This will be fulfilled using ELISA-based salivary ZG16B screening validated with immunohistochemistry staining using tissue slides. The completion of this project will provide rigorous training in biochemistry, salivary gland biology and saliva-based diagnostics enhanced in an interinstitutional research agreement (NIH Graduate Program Partnership) between Indiana University School of Dentistry (IUSD) and National Institute of Dental and Craniofacial Research (NIDCR). This collaboration incorporates excellent mentorship in clinical and translational research to prepare the applicant for a career as a dentist-scientist.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Infants and young children (IYC) make up nearly 50% of all pediatric Hematopoietic Stem Cell Transplants in the U.S. each year. HSCT is a high-risk treatment that requires lengthy hospitalization (min. 4 weeks), repeated invasive procedures, and high symptom burden. IYC (< 3 years) and parents experience sustained and severe distress during HSCT, with as many as 80% of children and 56% of parents reporting moderate to severe post-traumatic stress symptoms (PTSS) after treatment. Developmentally, IYC cannot effectively self- regulate and rely on parents to manage and buffer the impact of stressful events. High parent distress makes this buffering process less effective, leaving IYC vulnerable to unrelenting stress and medical trauma. Despite the significant and prolonged symptom distress experienced by IYC and their parents, there is a glaring absence of empirically validated interventions for this age group. To fill this gap, the overall scientific objective of this application is to adapt and evaluate an existing music-based intervention (Active Music Engagement; AME) to be developmentally appropriate for IYC and the HSCT treatment context, to promote parent buffering behaviors and decrease distress for both the IYC and their parent. The central hypothesis is that the adapted AME intervention will produce a clinically meaningful decrease in distress during HSCT in IYC and their parent. We will accomplish this research across 2 separate but related phases. In the K99 phase: First, (Aim 1) we will describe patterns of parent buffering behaviors, IYC distress, and parent distress across HSCT admission using a multi-case approach and parent interviews (n=6 parent/IYC dyads). Then, (Aim 2a) we will adapt the AME intervention based on existing research, relevant theoretical constructs, and findings from Aim 1. Finally, (Aim 2b) we will further refine AME through a series of n-of-1 optimization studies. In the R00 phase, we will conduct a small two-group randomized controlled pilot trial (n= 16 parent/IYC dyads) to (Aim 3) evaluate the feasibility and acceptability of the adapted AME and attention control condition for parent/IYC dyads during HSCT; and (Aim 4) determine if the adapted AME produces minimally clinically important differences (MCIDs) on measures of parent/IYC distress and that the attention control condition does not produce MCIDs. The overall goal of the mentored training program is to position Dr. Harman to have the skills needed to become an independent investigator in behavioral oncology with a focus on music-based interventions to mitigate distress and medical trauma in IYC and their parents. The proposed training plan encompasses four key areas: (1) developmental and trauma-related theories, (2) intervention development and trial design, (3) advanced statistical modeling and dyadic analysis, and (4) multi-site trial management. Findings will significantly impact our understanding of parent buffering behaviors and the distress experienced by parents and IYC during HSCT and provide an empirically tested intervention currently unavailable for our youngest patients at risk for long-term adverse health outcomes related to sustained and escalating distress.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract Tooth loss is still a public health concern, and it affects daily oral functions thereby impacting general health. Even though tooth loss can be rehabilitated through dentures, chewing efficiency declines compared to natural dentition, affecting one’s nutritional intake. Individuals who have lost all their teeth or partially dentate with and without prostheses are known to be at risk for malnutrition. As a result, it is essential to monitor their nutritional profile among adults experiencing edentulousness and receiving prosthodontic treatments. However, this is not a common procedure during dental visits. Although, many malnutrition screening (MS) tools exist in general health settings they are not used in dental practices because of lack of evidence for the tools’ performance, on the usability, and lack of understanding of patient outcomes among individuals with dentures. The long-term goal for our proposed research study is to facilitate a patient-centered healthcare system by translating the best scientific evidence into clinical practice through clinical decision support systems (CDSS), especially those that emphasize continuity of care between dental and other healthcare providers. The objective of this career development award is to evaluate the accuracy and performance of the three malnutrition screening tools, Nutrition Screening Initiative (NSI), Mini Nutritional Assessment-Short Form (MNA-SF), and Malnutrition Screening Tool (MST), and use them to screen and refer to a clinical registered dietitian, and to evaluate the feasibility of the tools’ implementation in dental practices. The rationale for this proposed work is to identify and refer denture-wearing patients at risk for malnutrition, and to reduce the risk for experiencing adverse effects of malnutrition which would compromise their quality of life. To accomplish the goal, I propose the following three specific aims: 1) Evaluate the usability of three MS tools with end users (denture wearers) and assess the tools’ accuracy in identifying malnutrition risk by comparing their results to a gold standard: a registered dietitian’s nutritional assessment. 2) Determine dental clinicians’ opinions of the three malnutrition screening tools and their perceived facilitators and barriers to implementing these tools in dental practices. Focus groups with DCs will collect information on their perspectives, opinions, knowledge, attitudes, and recommendations on using the MS tool for clinical decision-making by using the consolidated framework for implementation research (CFIR). 3) Evaluate the feasibility and effectiveness of implementing in dental practices the three malnutrition screening tools for screening and referring patients who receive/wear dentures. Iterative usability testing with denture wearers, clinicians, and contextual inquiry (CI) observations in dental practices will be used to understand the ease in use of the tool by the end user within the clinical workflow. The outcome of this research is to provide scientific evidence on the effectiveness of using malnutrition risk screening with denture wearers in dental practices. The proposed work will inform on the design and implementation of MS tools.

NIH Research Projects · FY 2025 · 2025-06

Project Summary Mosquito-borne illnesses, which are caused by pathogens that are spread through the bites of infected mosquitoes, are among the most significant infectious diseases worldwide and among the most challenging to prevent and control. The long-term goal of our research program is to combat mosquitoes through the transfer of eco-friendly RNAi-based mosquito gene silencing from the bench to the field, resulting in the development of a new class of eco-friendly pesticides to combat insecticide resistance. The proposed investigation will test the hypothesis, which is supported by strong preliminary data, that interfering RNA targeting vital mosquito genes, which has been produced in Saccharomyces cerevisiae, can be effectively delivered to Aedes, Anopheles, and Culex mosquitoes in soda, resulting in high levels of mosquito mortality. The objective of the proposed study is to develop the yeast endless soda (YES) feeding station, an innovative soda bottle-based system for deployment of RNAi yeast insecticides as attractive targeted sugar baits (ATSBs). Preliminary studies suggest that the yeast-soda combination is more attractive to mosquitoes than existing sugar bait lures, enabling it to better lure mosquitoes away from feeding on natural sugar sources. Moreover, unlike insecticides that are presently being deployed in other ATSB systems, the RNAi yeasts to be evaluated in the proposed investigation are specific to mosquitoes, posing little if any threat to non-target organisms. Soda in plastic bottles and yeast are inexpensive and readily distributed worldwide, making these reagents excellent selections for the development of these innovative ATSB bait stations. The project plan includes the following aims: 1) Generation of an eco-friendly soda and RNAi-based yeast ATSB as a universal mosquito lure that selectively kills mosquitoes and 2) Development and evaluation of a prototype YES feeding station. The expected outcome is the development of a consumer accepted ATSB station that can be further evaluated in field trials and eventually deployed globally to reduce the worldwide mosquito burden.

NIH Research Projects · FY 2026 · 2025-06

T follicular cells (Tfh) act as effector helper cells that are critical in B cell class switching into IgE producing plasma cells. Mast cells (MC) express the high affinity IgE receptor, FcεRI, that when complexed with IgE and antigen, lead to degranulation and the release of preformed mediators irrevocably tied to anaphylactic reactions. Learning more about Tfh and MC function and development can provide knowledge on new therapeutic targets for unmet clinical needs. TGFβ is a complex cytokine that plays a well appreciated role in the immune system, however the effects of TGFβ signaling disruption on IgE production and responses and how this may impact allergic disease is still poorly understood. Given the potent and diverse role of TGFβ, a better understanding of this pathway and its relationship to allergic disease pathogenesis is warranted. To clarity this, we study Tfh and MC function in patients with Loeys-Dietz Syndrome (LDS), an autosomal dominant disorder caused by mutations in TGFBR1 and TGFBR2, who are highly predisposed to food allergy, asthma, allergic rhinitis and eczema. Using a mouse model of LDS with a knock-in allele (Tgfbr1WT/Mut ) of an LDS mutation and patient samples, we found that LDS patients and mice have decreased lymphocyte pSmad2/3 signaling downstream of TGFβ, increased Tfh2 cells, decreased Tfh17 cells, and increased food specific IgE. LDS Tfh cells displayed a reduced STAT3 and SOCS1 signaling signature. Furthermore, LDS T cells demonstrated increased PI3Kγ transcription and activity, which could be replicated with siRNA knockdown of the TGFβ dependent transcriptional repressor, SnoN, and prevented with chemical inhibition of PI3Kγ. In support, in vivo PI3Kγ blockage resulted in decreased Tfh accumulation and IgE production in response to fed OVA antigen. Furthermore, LDS mice were unexpectedly protected from IgE mediated anaphylaxis. In addition, cultured mice and human LDS MC’s displayed suppressed IgE-mediated degranulation compared to healthy controls. Mechanistically, IL-33 responses were found to be enhanced in LDS MCs, which was tied to decreased IgE mediated functions since LDS mice deficient in the IL-33 receptor displayed a partially corrected anaphylaxis phenotype. Furthermore, transcriptome analysis revealed that the neuropilin (NRP) signaling pathway was enhanced in LDS MCs. MC specific NRP1 knockout (cKO) led to enhanced IgE driven anaphylaxis, suggesting a negative regulatory role for NRP1 in MC mediated IgE functions. Therefore, this project will test the hypothesis that TGFβ signaling suppresses Tfh2 development via PI3Kγ and SOCS1-STAT3 but augments MC IgE responses by blocking the negative feedback loops via ST2/IL-33 and NPR1.