Temple Univ Of The Commonwealth

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Childhood apraxia of speech (CAS) is a pediatric speech sound disorder (SSD) that impacts between 60,000 and 780,000 children under 14 in the US. The underlying impairment in speech motor planning primarily affects articulation and prosody and reduces intelligibility. Beyond impaired communication, children diagnosed with CAS are also at higher risk for difficulties with social/emotional development and reading and writing. Thus, effective targeted intervention is warranted for these children. However, CAS diagnosis is difficult and at present relies primarily on the `gold standard' clinical judgment by an `expert speech-language pathologist (SLP)' based on a number of perceptual features: (a) Inconsistent errors, (b) impaired transitions between sounds and syllables, and (c) impaired prosody, in particular lexical stress. This diagnostic standard is problematic for several reasons, including the fact that these features are not operationalized (in terms of elicitation, measures, number of instances required for diagnosis), lack precision, and vary with the SLP's language and clinical experience. These challenges are exacerbated by the well-known heterogeneity of CAS presentation and the overlap with other SSDs. Finally, there are no standards or criteria by which an SLP is considered an `expert'. Together, these major limitations lead to disagreements between SLPs and reluctance by some SLPs, especially less experienced ones, to diagnose CAS. This undermines the ability to make well-informed evidence-based clinical decisions for children with CAS and other SSDs. In addition, the research literature on CAS has been hampered for decades by this diagnostic problem, and is compounded by small sample sizes. More objective, validated, precise, replicable, and clinically feasible diagnostic measures are sorely needed. Acoustic measures have strong potential to meet these needs, based on prior small-scale studies in CAS and studies with other populations with speech disorders. However, to date no studies have systematically examined acoustic measures of core perceptual CAS features in a single study. This F31 proposal leverages a unique, large and well-characterized retrospective dataset of 125 children (76 with CAS, 28 with another SSD, and 21 without any SSD) to systematically examine the validity of a well- motivated set of acoustic measures of each core perceptual feature (3 acoustic measures each) (Aim 1), the differential diagnostic accuracy of the three acoustic measures with highest validity (1 acoustic measure per feature) (Aim 2), and the potential identification of CAS subtypes based on these acoustic measures (Aim 3). As a first step, the diagnostic accuracy of these acoustic measures will be evaluated against a gold standard of an operationalized consensus criterion based on independent judgments from 3 expert SLPs. This F31 project will provide novel, unique, and significant information that will help improve clinical diagnosis of CAS and other SSDs, provide the Fellow with advanced training to transition toward an independent research career, and set the stage for a future prospective validation study of the acoustic measures found most promising here.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract The foundation of the Kim Research Group at Temple University is built upon the development of new designer reagents and methods to access high-value carbonyl compounds from acetal-based radical precursors. To that end, this proposal strives to implement acetal radicals as a means to access a diverse array of medicinally relevant molecular platforms. To address these challenges, we have designed a set of novel reagents to overcome rapid decarbonylation side reactions of acyl radicals and developed a platform to synthesize carbonyl compounds of interest to those in the agrochemical, pharmaceutical, and material industries. Additionally, our fundamental research in acetal radicals has provided alternative opportunities to study them in fragment coupling reactions. This work will aid in the rapid construction of molecular complexity by combining C–H activation with C–C bond formations to construct a wide variety of molecular scaffolds. This research is well poised to tackle necessary synthetic challenges in order to build pharmaceutically relevant precursors, allowing it to be widely adopted within the synthetic medicinal community. We have devised two research programs to address these research aims. Research Program I will explore the use of acetal radicals to install high-value ketones that can serve as carboxylic acid bioisosteres. Research Program II will explore alternative C–C couplings of acetal radicals to install highly oxidized carbon fragments for complex molecular syntheses. This research will be essential in shaping the next generation of design principles for improved pharmaceuticals to tackle human diseases more effectively.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The proposed K23 career development award in clinical and translational research will form a foundation for Dr. Yuan Liu to become an independent clinician-scientist, with expertise on severe early childhood caries (S-ECC). The K23 training objectives progress in a sequential fashion and will support her development of key expertise in 3 areas: 1) developing advanced skills in designing and conducting clinical studies; 2) acquiring advanced knowledge and methods in fungal biology and microbiome; 3) improving scientific communication, grant writing skills and networking. The candidate will acquire experience in directing longitudinal study and essential training by making the use of the extensive resources at the University of Pennsylvania and capitalizing on an interdisciplinary mentoring committee with broad, complementary expertise in clinical research, epidemiology, bioengineering, microbiome, and bioinformatics. S-ECC is a major public health problem characterized by dysbiotic oral microbial burden leading to a persistent and virulent biofilm on the teeth of toddlers that causes rampant tooth decay. This process not only destroys the dentition but can also have a negative impact on the general health, such as complicated systemic infections, and the well-being of young children. Emerging clinical evidence has spotlighted the potential role of Candida albicans in S-ECC. Furthermore, our retrospective study using electronic health records (EHR) from six major children’s hospitals revealed that, oral thrush (oropharyngeal candidiasis or OPC) detection in toddlers less than 12 months of age was strongly associated with the development of dental caries soon after, particularly between 13 and 36 months. However, only cross- sectional human studies have been performed thus far. Longitudinal studies are warranted to better assess the causal association between OPC and S-ECC in toddlers. In parallel, further studies are needed to determine the fungal role in caries and if Candida colonization in oral cavity could be a reliable clinical marker for risk of S-ECC. To address this, the proposed research will be focused on conducting a 2-year prospective longitudinal study to investigate the association between early OPC and the progression of S-ECC, and the impact of Candida colonization on the functional plaque microbiome and biofilm virulence. Dr. Liu will investigate 3 specific aims: (1) examine the association between early OPC and S-ECC development in a cohort of infants; (2) evaluate the influence of OPC and Candida colonization on the plaque microbiome in infants; (3) study the role of Candida on biofilm cariogenic properties. Results from this clinical and translational effort will further elucidate the causal link between OPC and S-ECC, and enhance the understanding of fungal contributions to the etiopathogenesis of S-ECC. Collectively, these results will provide future opportunities for mechanistic clinical studies and developing targeted strategies for preventing S-ECC. This K23 award will provide Dr. Liu with new knowledge in the areas essential to her career goals as a clinician-scientist, implement the proposed research plan and build an independent research program with the goal of receiving an R01 prior to the end of the K23 award.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Confirmation bias refers to the tendency to interpret new information in a way that conforms to one's pre-existing beliefs. It is associated with deficits in cognitive functions like working memory and error detection, both commonly associated with the prefrontal cortex (PFC). As such, age-related changes to the PFC may leave older adults vulnerable to this bias and its negative outcomes, including financial exploitation. Older adults are disproportionately impacted by financial exploitation, and as cognitive decline progresses to mild cognitive impairment (MCI) and Alzheimer's disease and related dementias, risk for fraud increases. Given these connections, confirmation bias may serve as an indicator of cognitive decline and vulnerability to fraud; however, progress towards understanding these relationships has been hampered by three main issues. First, little is known about the latent processes shaping confirmation bias in older adults. Second, much of the work to date has relied on correlations between brain responses and behavior, leaving the causal mechanisms unclear. Third, behavioral changes over time in this population are underexplored. To address these concerns, this project has two specific aims. The first is to characterize how age-related differences in cognitive function contribute to susceptibility to confirmation bias. In an online study, participants will play an investment game involving the opportunity to update an initial judgment after receiving feedback from a partner (Aim 1). We hypothesize that older adults showing cognitive decline will exhibit confirmation bias at higher rates than younger adults, characterized by greater bias towards initial judgments and decreased engagement with conflicting feedback from the partner. Our second aim is to assess the causal relationship between confirmation bias and the PFC in older adults using transcranial alternating current stimulation (tACS; Aim 2). By applying tACS to the PFC, we will entrain individualized theta rhythms to enhance function across the region. Using the investment task from Aim 1 concurrent with tACS, it is hypothesized that confirmation bias will be diminished in older adults, particularly among those with MCI. Finally, to fully characterize the predictive role of confirmation bias, we will follow up with participants after two years to assess the progression of cognitive decline. It is predicted that higher rates of confirmation bias at visit one will be associated with greater cognitive decline at follow-up. The funding from this grant will provide the applicant with the opportunity to learn essential new skills in computational modeling (Aims 1 & 2) and noninvasive brain stimulation (Aim 2). These objectives will be accomplished through a combination of workshops, expert mentorship, and institutional training. Overall, this project will leverage training in well- established and cutting-edge techniques to help expand the understanding of cognitive decline and provide insight into causal links between PFC function and susceptibility to confirmation bias. More generally, it may provide new insights into reducing older adults' susceptibility to the bias and their risk for financial exploitation.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Youth at risk of HIV in the US are less likely to be aware of, use, or adhere to PrEP. This is especially worrisome as new modalities beyond daily oral (i.e. injectable, longer acting pills) are quickly coming on the market, making decision making and clarifying preferences crucial to uptake. Importantly, missing in much of the research on barriers to PrEP uptake are the voices of young (15-24) people at risk for HIV, who may experience age-related barriers to PrEP decision making, such as confidentiality, conflicts with family, and lack of a supportive community with credible spokespeople whom they trust. No interventions have developed decision support tools (DSTs) to help youth at risk for HIV make decisions about PrEP in line with their individual preferences and values that meet youth where they are. The primary goal of this study is to develop and evaluate the feasibility, acceptability and promise of efficacy of an electronic health DST called QUEEN-P (QUestions to Empower and ENgage with PrEP) to increase PrEP informed decision making among youth at risk for HIV in Philadelphia. Using commercial marketing techniques – perceptual mapping and vector message modeling – and grounded in the Ottawa Decision Support Framework, QUEEN-P will be delivered via tablets or other devices in novel locations (mobile vans, events), working with community partners known to be trusted by youth. Specific aims are: 1. Assess perceptions of PrEP, different delivery methods and potential barriers to initiation and adherence. We will establish community and scientific advisory boards (CAB/SAB) made up of youth and healthcare providers and community advocates, interview youth (n=20) to assess barriers, survey youth (n=100) and use perceptual mapping analyses to develop targeted messages, and elicit feedback from the CAB/SAB on content and delivery of QUEEN-P; 2. Develop the QUEEN-P eHealth DST and assess usability and acceptability. We will develop a prototype DST and concept-test messages and features with our CAB/SAB for feedback on acceptability, revise it based on feedback and conduct user testing (n=20); 3. Pilot the intervention to assess for promise of efficacy in engaging youth at risk for HIV in PrEP informed decision making and assess interest in and outreach to community clinical partners for PrEP. We will conduct a pilot test (n=60) in Philadelphia with youth at risk for HIV randomized to an “Enhanced” (QUEEN-P + navigation to PrEP providers) or “Basic” study arm (basic PrEP information and a list of resources). To assess promise of efficacy, we will compare outreach to clinical partners and decisional conflict (primary outcomes) at 3-month follow-up and assess decision preparation, self-reported knowledge and attitudes, and PrEP uptake (secondary outcomes; immediate post, 3 month follow-up). Qualitative exit interviews will inform feasibility and acceptability. This pilot study will examine the potential impact of using technology-based interventions to improve PrEP informed decision making in a population at high risk of HIV acquisition.

NIH Research Projects · FY 2025 · 2024-08

PROJECT ABSTRACT Added sugar (AS) is a critically important dietary target for intervention in adolescents; it contains excess calories with no nutritional value and is independently associated with chronic disease risk. Adolescents consume more AS than any other age group and are at a unique phase in the life course where improving diet could have a significant influence on overall health into adulthood. However, capturing dietary change remains a paramount challenge in the field of nutrition and this is particularly true for overconsumed nutrients like AS that are highly susceptible to reporting error and social desirability bias. The challenge of dietary self-report is also pronounced in behavioral trials where issues like participant burden, repeated measures, and small sample sizes have significant impacts on the ability to accurately assess outcomes. The carbon stable isotope ratio (CIR) is a low-burden and valid measure of AS and sugar-sweetened beverage (SSB) intake that warrants additional investigation in dietary interventions. Most of the evidence on CIR to date is derived from controlled feeding studies and observational trials using blood and hair CIR. Breath CIR has recently been validated as a measure of short and long-term AS intake and provides an exciting new avenue to explore AS intervention adherence and effectiveness in behavioral trials. Thus, the purpose of this R03 is to examine CIR from breath as an indicator of intervention adherence (i.e., short-term intake) and intervention effectiveness (i.e., change over time). Additionally, this study aims to assess the differential feasibility, acceptability, and sensitivity to change among CIR from breath, blood, and hair samples to inform work in future trials. This R03 study will be mapped onto a randomized controlled trial (RCT) in an ongoing K01 study. Adolescents (n=70) 12-16 years will be randomized to a 12-week behavioral intervention to reduce AS intake (n=35), or a nutritional education comparison (n=35). This R03 study will 1) Assess the feasibility and efficacy of using daily breath CIR (fasting morning, postprandial afternoon, and evening) to capture intervention adherence via repeated sampling over 12-weeks (at baseline, and in weeks 3, 6, 9, and 12); 2) Examine AS intervention effectiveness based on changes in breath CIR from baseline to end-of-intervention (12-weeks) between intervention and control participants; and 3) Compare feasibility, acceptability, and sensitivity to change of breath, blood, and hair CIR to capture change in AS over 12 weeks. This R03 will enhance the impact of the PI's existing K01 by adding innovative breath CIR measures and analyses, which will provide robust evidence for objectively assessing AS intervention adherence and effectiveness. Additionally, this study will test assessment approaches that could reduce participant burden and eliminate reliance on dietary self-report, resulting in more efficacious strategies for populations at greatest risk of diet-related health disparities. Findings will facilitate the PI's research trajectory to advance the science of dietary assessment methodology and expand the evidence for the use of objective biomarkers in behavioral nutrition interventions.

NIH Research Projects · FY 2026 · 2024-08

Project Summary Eosinophilic esophagitis (EoE) is a chronic food allergen- and immune-mediated disease that exerts a significant clinical and financial burden worldwide. Interleukin (IL)-13 is an essential mediator of EoE due its pleotropic effects on esophageal inflammation and tissue remodeling. We have identified IL-13 as a critical mediator of increased mitochondrial mass and mitochondrial DNA (mtDNA) release in esophageal epithelial cells. Our preliminary data indicate that IL-13-mediated alterations in mitochondrial mass and mtDNA release are dependent upon JAK/STAT signaling. We further provide preliminary data suggesting that depletion of mitochondria limits EoE-associated epithelial remodeling and inflammation. These findings support our overarching hypothesis that IL-13 in the EoE inflammatory milieu activates STAT signaling in esophageal epithelium to drive alterations in mitochondrial biology that promote EoE pathogenesis and may serve as novel biomarkers in EoE patients. To test this hypothesis, we will define the direct molecular mechanisms through which IL-13 signaling regulates mitochondrial biology (Aim 1) and the functional significance of these findings with regard to EoE-associated tissue remodeling and inflammation (Aim 2). Furthermore, we will elucidate the clinical significance of EoE-associated alterations in mitochondria by exploring their biomarker potential in EoE patients (Aim 3). The proposed studies will illuminate the IL-13/STAT/mitochondria axis as a novel player in EoE pathobiology and have great potential to improve clinical care in this disease that remains a major cause of upper GI morbidity.

NIH Research Projects · FY 2024 · 2024-08

Project Summary Ambient air pollution is a major pediatric public health concern due to its ubiquity and increased exposure patterns affiliated with climate change over the next decade. Despite significant improvements in overall levels of air pollution over the 20 years, 62% of children still live in areas that do not meet the US Environmental Protection Agency National Ambient Air Quality Standards. Consistent evidence has shown that ambient air pollution exposure is detrimental to children's respiratory health due primarily to behavioral patterns and timing of exposure during a sensitive window of respiratory, endocrine, and immune system development, maturation, and modulation. Given the growing risk of present and future pandemics, and consistent exposure to environmental stimuli, understanding the role of air pollution on pediatric respiratory disease is of pressing public health importance. While overall distributed and respiratory Minoritized poorer respiratory data pediatric discharge project Statewide estimates enrollment respiratory support using characteristics neighborhood-level aims young children in the United States have a high burden of acute r espiratory diseases, r espiratory related hospital admissions are not equally across racial and ethnic populations. Rates of hospitalization are consistently greater among Black Latinx populations. These observed r acial and ethnic inequalities often remain after adjustment for traditional risk f actors but are reduced when environmental and sociodemographic factors are considered. and low-income populations are more likely to reside in areas with greater levels of air pollution, quality built environment, and higher neighborhood deprivation, all factors which may impact acute disease in children. To bridge these gaps in knowledge, the proposed work will capitalize on existing to examine the complex relationships between air pollution and neighborhood-level characteristics on acute respiratory disease. The proposed study is a secondary data analysis of administrative hospital data to examine environmental risk factors for pediatric acute respiratory disease. The proposed will utilize participant data that has already been collected as part of the New York Department of Health Planning and Research Cooperative System (SPARCS) dataset. The addition of air pollution and neighborhood-level characteristics to the existing data does not require participant contact or new of participants. Specific outcomes of interest include asthma exacerbation, acute upper and lower infections, allergic rhinitis, and otitis media. This R03 will provide the protected time and programming o accomplish the following aims: (1) Examine the effect of air pollution on acute respiratory disease a case-crossover study design, (2) Determine the effect of objective measures of neighborhood level on acute respiratory disease, and (3) Using a mixture model, evaluate which highly correlated characteristics and air pollution metrics are associated with acute espiratory disease. All will assess whether these associations vary across age, race, ethnicity, and/or sex. t r Results from the proposed work will enhance the current knowledge by understanding the combined effect of air pollution and neighborhood-level exposures on respiratory disease in a generalizable sample of children

NIH Research Projects · FY 2025 · 2024-08

Voice disorders occur in 6–17% of children, resulting in dysphonia (i.e., altered vocal quality), which has significant negative health, social, emotional, and educational consequences if left untreated. Voice therapy is the gold-standard treatment for children with vocal fold nodules, the most commonpediatric voice disorder. Most children diagnosed with vocal fold nodules receive therapy at one of the few specialized pediatric voice clinics in the country. Families that do not have access to these specialized clinics may turn to the school system for therapy, however, speech-language pathologists (SLPs) in the school system self-report lower competency levels and lack of knowledge and comfort in treating pediatric voice. Therefore, the scarcity of specialized pediatric voice clinics has created a significant health equity issue as children unable to attend specialized pediatric voice clinics due to lack of financial or physical access will be significantly disadvantaged compared to their more affluent and well-situated peers. To address this critical gap, the long-term goal of this research program is to create training programs and materials for SLPs who practice outside of specialized clinics (e.g., the schools), with materials based on the best practice derived from expert pediatric voice clinicians. This research program is designed to address multiple priority areas of the NIDCD, including promoting health equity (Theme 4) and scientific advancement (Theme 5). Outcomes will also address the NIH-Wide Crosscutting Priority of reducing health disparities. The current study will identify key themes of expert pediatric voice therapy, a critical first step to improving the knowledge and skills about pediatric voice therapy for SLPs in all settings. Outcomes from this work will identify best practices in expert pediatric voice therapy, providing an essential foundation for a subsequent R01 to develop a training program for treating pediatric voice. In the current work, pediatric voice therapy sessions will be videotaped without interference at two top, high-volume pediatric voice centers (Boston Children’s Hospital, Cincinnati Children’s Hospital Medical Center). Video recordings will be analyzed offline using a two-phase explanatory sequential mixed method design, with the quantitative analysis phase followed by a qualitative analysis phase. Aim 1 will use qualitative content analysis to examine communication methods (relational, instrumental, affective) based on interpersonal communication theory. Aims 2 and 3 will be data-driven thematic analyses of how experts structure therapy sessions, such as the task order or how to incorporate games and breaks (Aim 2) and how experts implement frequently used therapy tasks (Aim 3). The research will be completed at Temple University (R1 university), which maintains a robust research environment with exceptional dedication and support to early-career faculty. This Early Career Stephen I. Katz R01 submission represents two clear changes in direction for the PI, a change in methodology (mixed methods design, qualitative analyses) and a change in population (expert pediatric voice clinicians).

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The neglect of children and adolescents, a form of maltreatment along with abuse, is a significant problem that impairs health and well-being throughout the lifespan. The long-term goal of this exploratory/developmental research grant (R21) is to lay the groundwork for a larger research grant (R01) to examine the multilevel antecedents of adolescent neglect, information that is critical to the development of interventions. The objective is to conceptualize, develop and initially validate a multidimensional youth self- reported measure of adolescent neglect. Neglect is conceptualized as inadequately met adolescent needs that present actual or potential harm. This shifts from previous definitions which have focused on parental omissions in care. This adolescent-centered conceptualization helps reconcile neglect's definition with growing scientific evidence of its complex multilevel etiology. By untangling unmet needs from parental omissions, the adolescent-centered conceptualization allows for more rigorous research examining neglect's multilevel antecedents. Importantly, it offers a more constructive frame that should improve parent and youth engagement in intervention. Aim 1 is to conceptualize, develop and pilot test a youth self-report measure of adolescent neglect, defining it as critical unmet adolescent needs. It will apply rigorous qualitative methods per state-of-the-art Patient-Reported Outcome Measurement Information System (PROMIS) scientific standards for establishing content validity. Guided by an initial conceptual framework, this will include a scoping literature review, a two-round modified Delphi expert panel (N = 12) and cognitive and concept elicitation interviews with adolescents (N = 18). The second aim is to establish initial psychometric properties of the new neglect measure. We will explore the measure's dimensional structure and assess its reliability (internal consistency, test-retest), validity (criterion, concurrent and predictive convergent), and invariance in a sample of 12-17-year- olds in Philadelphia who are at risk for neglect (N = 400). The outcome of the proposed research will be a rigorously developed, novel measure of adolescent neglect with a sound conceptualization that incorporates input from diverse experts and youths (Aim 1) and evidence of psychometric properties (Aim 2). The measure has the potential for significant impact, paving the way for a new generation of neglect scholarship with a sound conceptual and operational basis that will illuminate the scope, multi-level contributors, and sequelae of adolescent neglect, informing policy and intervention that can overcome this intractable and pernicious problem.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Degeneration of neuronal axons is a hallmark of diverse neuropathological conditions, ranging from acute nerve injuries to chronic neuropathies and neurodegenerative diseases. If we better understood the mechanisms of axon degeneration, we could devise new therapies to potentially slow or prevent axon degeneration and ameliorate a range of neuropathologies. Recent findings suggest that one key event in axon degeneration is the loss of ‘axon survival factors’, labile proteins whose continuous supply from neuronal cell bodies to distal axons normally ensures axon integrity. The damage to axons that occurs after injury, or in chronic neuropathies, disrupts the supply of survival factors and triggers distal axon degeneration. The best-known axon survival factor, nicotinamide mononucleotide adenylyl transferase-2 (NMNAT2) requires the protein-lipid modification palmitoylation for its fast axonal transport, but how palmitoylation contributes to NMNAT2 degradation is not fully clear. I identified previously unrecognized phosphorylation sites on NMNAT2, whose phosphorylation by c-Jun N-terminal kinase (JNK) triggers selective degradation of palmitoyl-NMNAT2 in cultured cells. While addressing this question, I also realized that several other regulators of axon degeneration are palmitoylated, in addition to NMNAT2. Given that palmitoylation is the only reversible protein-lipid modification, I asked if acute depalmitoylation of NMNAT2 and/or other axonal proteins is important for axon degeneration. My results revealed that pharmacological inhibition of protein depalmitoylases indeed protects axons from degeneration and additional preliminary genetic data ascribe this protection to block of a recently described depalmitoylase, ABHD16A. However, axon protection by depalmitoylase inhibitors is independent of NMNAT2 and other known axon survival factors, suggesting that additional protein(s) are depalmitoylated after axonal damage to drive subsequent degeneration. The hypothesis that underlies this proposal is that palmitoylation plays broad, previously unrecognized roles in the control of axon degeneration. Experiments in Aim 1 will use a recently generated phospho-mutant knockin mouse to define the importance of phosphorylation by JNK for NMNAT2 degradation and subsequent axon degeneration, both in cultured neurons and in vivo. Aim 2A will determine whether ABHD16A is indeed the key depalmitoylase whose activity is required for axon degeneration and will provide the first cell biological characterization of ABHD16A localization in axons. In Aim 2B I will break new ground by comprehensively identifying axonal proteins that are rapidly depalmitoylated after axonal injury, in order to identify those protein(s) whose acute depalmitoylation drives axon degeneration. Together, the results of these studies will provide new insights into the cell biology of axonal (de)palmitoylation, will define key events that drive axon degeneration, and could reveal new therapeutic strategies to ameliorate a host of neuropathological conditions.

NIH Research Projects · FY 2024 · 2024-08

The goal of this proposal is to fully delineate the molecular mechanisms by which the prominent serine/threonine Protein Phosphatase 2 (PP2A) holoenzyme, specifically PP2A/B55α, recognizes its substrates through a new helical motif and to use this information to identify direct PP2A/B55α substrates with functions in mitogenic signaling and early cell cycle progression. Protein phosphorylation is an essential regulatory mechanism in cell signaling and the regulation of the cell cycle and it is altered in cancer, Alzheimer, and other diseases. A large fraction of dephosphorylation is mediated by distinct PP2A trimeric holoenzymes. These consist of a scaffold protein that connects the catalytic subunit to the variable regulatory subunit, which mediates substrate specificity. The most abundant holoenzyme is PP2A/B55α. Its activity has been implicated in mitogenic signaling and early cell cycle progression, but details remain unclear. Initially, it was believed that PP2A/B55α recruited substrates through charge-charge interactions between the surface of B55α and substrates. However, our groundbreaking discoveries challenged this notion. We identified a conserved amino acid motif in various proteins, including the p107 substrate and the FAM122A inhibitor, that folds as an a-helix. This helical motif is essential for p107 dephosphorylation and for FAM122A binding via similar interactions with residues in a groove on the top of B55α. This mechanism is distinct from that used by other Ser/Thr phosphatases that bind short linear motifs in intrinsically disordered protein regions. Gap: Despite this advance, we remain unable to identify other PP2A/B55α substrates using this short helical sequence. This is due to the lack of a comprehensive assessment of amino acid variability within the helical motif sequence and incomplete detail of the motif’s structural plasticity. As a result, it remains extremely difficult to identify new substrates, to distinguish correctly vs incorrectly assigned substrates, and to demonstrate the veracity of correctly assigned substrates. We hypothesize that a helical region with a defined length and a variable conserved amino acid sequence in PP2A/B55α substrates mediates binding to the top groove of B55α via specific electrostatic and hydrophobic interactions. This is a primary mechanism for substrate recognition that modulates signaling through the cell cycle. These will be tested with two aims: (1) To define PP2A/B55α substrate specificity via a short helical motif with defined sequences. (2) To dissect the PP2A/B55α substrate interplay in mitogenic signaling linked to the cell cycle Restriction Point (RP). DEIA Aims: Diversity among Temple University undergraduates is excellent, but the transition of underrepresented minority (URM) students to biomedical science research careers or graduate schools is still severely limited. Our goal is to implement a program we have been piloting that is geared toward eliminating barriers to joining the scientific research workforce faced by URM undergraduates at Temple and nationwide. The program will recruit 15 students who will work in mentors’ labs on a specific project and receive programmatic training to prepare them for admission and success in Biomedical Research PhD programs. Aim 1 is to develop research skills and knowledge and Aim 2 is to develop career skills by providing instruction in fellowship writing, graduate school application, oral and writing scientific communication, and scientific networking. Successful completion of the scientific Aims will facilitate identification of PP2A/B55α substrates and their validation in multiple pathways and cellular processes often deregulated in disease providing novel targets for therapeutic intervention. The DEIA training program will guide URM students to professional research careers.

NIH Research Projects · FY 2024 · 2024-08

Project Summary Eosinophilic esophagitis (EoE) is a chronic food allergen- and immune-mediated disease that exerts a significant clinical and financial burden worldwide. Interleukin (IL)-13 is an essential mediator of EoE due to its pleotropic effects on esophageal inflammation and tissue remodeling. We have identified IL-13 as a critical mediator of increased mitochondrial mass in esophageal epithelial cells. We further demonstrate that IL-13-mediated alterations in mitochondrial mass are dependent upon JAK/STAT signaling and contribute to EoE-associated epithelial remodeling. These findings support our overarching hypothesis that IL-13 in the EoE inflammatory milieu activates STAT signaling in esophageal epithelium to drive alterations in mitochondrial biology that promote EoE pathogenesis. To test this hypothesis, we will define the direct molecular mechanisms through which IL-13 signaling regulates mitochondrial biology (Aim 1) and the functional significance of these findings with regard to EoE-associated tissue remodeling (Aim 2). The proposed studies will illuminate the IL- 13/STAT/mitochondria axis as a novel player in EoE pathobiology. The research proposal, coupled with the tailored training plan, will provide outstanding career and professional development opportunities for the applicant under the guidance of the co-mentors who have relevant expertise in esophageal biology (Whelan) and mitochondria (Elrod).

NIH Research Projects · FY 2026 · 2024-07

Interpersonal violence (IV) has detrimental health impacts for all individuals, with the consequences of unaddressed victimization representing a key driver of poor mental and physical health among survivors. To progress from merely identifying these disparities to actively intervening, innovative interventions are needed to promote healing engagement, build bridges within survivor care infrastructure, and connect survivors with appropriate resources to help them heal. The long-term goal is to establish survivor-centered interventions that promote healing and wellbeing for survivors of interpersonal violence. The primary objective of this research is to refine and evaluate a photo-elicitation intervention, Photo-experiencing and Reflective Listening (PEARL), aimed at identifying help-seeking barriers and promoting healing engagement among IV survivors. The rationale for this project is that there is a critical unmet need for interventions that identify survivor-centered barriers and facilitators of trauma healing while creating a structure for survivors to identify their needs and co-create a healing action for themselves moving forward. This project has two key aims: 1) to refine the PEARL intervention prototype for implementation with men and women who have experienced IV, and 2) to evaluate the feasibility, acceptability, and potential impact of PEARL in survivor populations (N=40; 30 women, 10 men). The study will employ a community-engaged, mixed methods approach. Aim 1 will involve collaborating with a community and scientific advisory board to refine the PEARL prototype and enhance the effectiveness and applicability of PEARL. Aim 2 will employ a randomized waitlist control design to assess feasibility, acceptability, and PEARL's potential impact on healing engagement and related health outcomes. The research is paired with a career development plan that will equip the applicant with the skills necessary for community-engaged intervention development, universal design, and implementation science to transition her into an independent researcher that can improve the survivor care landscape through her program of research. The research proposed in this application is innovative because it focuses on recovery rather than deficit in survivor populations, using trauma-informed, and healing-centered approaches. It also leverages photo-elicitation, allowing survivors to highlight their multidimensional social world and reflect on how it might facilitate, create barriers, or shape healing needs in real time. This study's significance lies in its response to the pressing need for interventions to explore the barriers and facilitators of health that influence IV survivor healing, all while promoting survivor engagement in the healing process. By bridging existing gaps in IV survivor support, this research aligns closely with NINR's mission to promote and identify factors that promote health and wellbeing in populations exposed to trauma and violence.

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract Cardiovascular disease is the leading cause of mortality in the US and worldwide. Decades of endeavors in studying cardiovascular disease have led to a substantial understanding of underlying mechanisms, but there is still much to be learned. Cardiovascular homeostasis is regulated at both transcriptional and posttranscriptional levels, and an increasing body of evidence suggests that RNA posttranscriptional modifications, such as alternative RNA splicing, play essential roles in regulating cardiac function and disease. A previous study by the applicant (Dr. Peiheng Gan) has revealed that RNA-binding protein with multiple splicing (RBPMS) is crucial for cardiomyocyte proliferation and heart development through modulating alternative RNA splicing. In the new study, the applicant found that RBPMS is required for adult cardiac contractility. The absence of RBPMS impaired cardiomyocyte contractility and impacted the splicing of various sarcomeric genes, which displayed distinct patterns. Cardiac RBPMS expression was decreased in patients with heart failure and doxorubicin-induced heart failure in animal models. Intriguingly, the overexpression of RBPMS showed protective effects on contractility and survival in doxorubicin-treated human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Together, these data suggest that RBPMS is a key regulator of adult heart function and heart failure progression. During the K99 phase of this proposal (Aim 1), the applicant will characterize how RBPMS regulates cardiomyocyte contraction through modulating RNA splicing using both mouse models and hiPSC-CMs. In Aim 2, the applicant will leverage cellular and molecular tools to characterize the functional interactions between RBPMS and other cardiac RNA binding proteins (RBPs), including RBM20 and RBFOX1. This study provides the applicant with a unique opportunity to investigate how different RBPs interact to regulate cardiac function and alternative RNA splicing. During the R00 phase (Aim 3), the applicant will explore the therapeutic potential of overexpressing RBPMS in doxorubicin-induced mouse heart failure and determine the molecular mechanisms of RBPMS-mediated protective effects. The completion of the proposed study will provide critical insights into the posttranscriptional regulation of cardiac function by RBPs and create new opportunities for curing cardiovascular disease. The applicant will acquire crucial knowledge and skills by studying RBPs and RNA posttranscriptional modifications during his K99 phase to complement his previous expertise in cardiovascular research. Additionally, the applicant’s career development will be enhanced by the expertise of an exceptional mentoring and advisory committee, as well as the unparalleled resources and ample educational and training opportunities at UT Southwestern Medical Center, a world-class research institution. The outstanding mentoring, unmatched resources, and strong commitment from his department will strengthen the applicant’s candidacy for and transition to an independent tenure-track faculty position.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Social deficits are a primary characteristic of autism spectrum disorder (ASD). The social motivation hypothesis posits that these deficits reflect alterations in social information processing, including a blunted reward response to social stimuli. Neuroimaging research suggests this blunting may generalize beyond the social domain. Given the considerable resources devoted to motivation and reinforcement-based interventions, it is essential to identify the extent that blunted reward processing influences the entire spectrum of autism (i.e., beyond social motivation) and to identify the neural mechanisms that contribute to said blunting. Proposed aims make substantive progress towards answering these questions in neurotypical (NT; N=50) vs. autistic (N=50) youths. We focus on emerging adolescence (10-15 years; 20% female reflecting autism prevalence) as it is a critical phase of development when peer feedback gains salience in NT youth and social deficits increase in autistic youth. Significance is derived from three factors. First, we address two shortcomings in fMRI study design that have hindered progress towards isolating ASD-related differences in reward processing: A) Most studies fail to directly compare neural response to social (e.g., viewing a smiling face) and non-social (e.g., winning money) outcomes. B) Social stimuli typically used to study reward fail to model the experience of actually being liked by peers (i.e., direct social reward). To overcome these shortcomings, we developed an fMRI-based feedback paradigm that quantifies neural response to reward in social (being liked) and non-social (winning money) domains. Aim 1 uses this paradigm to isolate differences in neural response to social and non-social reward. This will be the first fMRI study to test ASD-related neural response to direct social reward. Second, we address a major knowledge gap in the social motivation hypothesis - the extent to which ASD symptoms relate to dopamine (DA) system function in brain. Animal models of autism suggest that DA system dysfunction contributes to blunted reward processing. However, testing human correlates has proved challenging because it requires invasive techniques involving radioactivity. Recent advances have demonstrated that neuromelanin-MRI (NM-MRI) can be used as a non- invasive proxy for measuring DA system function. Aim 2 quantifies differences in midbrain DA system function via NM-MRI. If awarded, this study will be the first to apply NM-MRI in ASD research. Third, in Aim 3, we lay the foundation for a novel approach to individualized treatment by testing the extent to which DA system function influences relations between ASD and blunted reward response. Gold standard interventions (e.g., Applied Behavioral Analysis, ABA) rely on intact reward processing for maximal effect. If DA system deficits diminish neural response to reward, this may explain variation in treatment outcomes. NM-MRI could identify those who may benefit most (or least) from ABA, and identify novel pharmacological targets to treat core symptoms of ASD.

NIH Research Projects · FY 2026 · 2024-06

SUMMARY This project aims to investigate the factors involved in lung epithelial regeneration following bacterial pneumonia. The regeneration process relies on the activity of stem/progenitor cells, particularly lung alveolar epithelial type 2 (AT2) cells, which differentiate into alveolar epithelial type 1 (AT1) cells. However, defects in this differentiation process occur in certain populations, such as the elderly and those with chronic respiratory diseases. Understanding the molecular mechanisms governing AT2 cell activity is crucial for developing effective therapeutic approaches. This project is based on the hypothesis that calcium-dependent metabolic changes drive epigenetic reprogramming, promoting the gene program required for AT2-to-AT1 cell differentiation. Specifically, the lysine trimethylation status of histone H3, controlled by Jumonji C (JmjC) domain-containing histone lysine demethylases (JMJDs), plays a key role in regulating gene expression and cellular differentiation. Pharmacological inhibition of JMJDs increased levels of trimethylated histone lysine marks and decreased AT2-to-AT1 cell differentiation, suggesting the necessity of histone lysine demethylation for AT2-to-AT1 cell differentiation. The activity of JMJDs is regulated by the succinate/α-ketoglutarate (αKG) ratio, which is influenced by mitochondrial matrix Ca2+ (mCa2+) levels. MICU1, a regulatory subunit of the mitochondrial Ca2+ uniporter channel, controls mCa2+ uptake from the cytoplasm into mitochondria and affects the succinate/αKG ratio. Our preliminary studies demonstrate that Micu1 deletion in AT2 cells decreased glutaminolysis feeding into the TCA cycle, increased succinate/αKG ratio and levels of histone lysine trimethylation marks. Additionally, supplementation with succinate increased histone lysine trimethylation marks and inhibited AT2-to-AT1 cell differentiation. Based on these findings, we propose that MICU1- dependent mCa2+ uptake regulates AT2 cell differentiation capacity by controlling the succinate/αKG ratio. This reinforcement of αKG-dependent JMJD activity promotes the gene program required for AT2-to-AT1 cell differentiation. This project has three specific aims. In aim 1, we will investigate how the succinate/αKG ratio connects MICU1-dependent mCa2+ uptake to AT1 cell fate during AT2-to-AT1 cell differentiation. Aim 2 will determine the mechanistic role of glutaminolysis in AT2-to-AT1 cell differentiation and lung epithelial repair. In aim 3, we will determine whether histone lysine trimethylation marks, regulated by MICU1-dependent mCa2+ uptake, control the gene program necessary for AT2-to-AT1 cell differentiation. Through unique mutant mouse models and clinically-relevant model systems, this project will provide valuable insights into the mechanisms underlying lung epithelial regeneration and contribute to the development of novel therapeutic strategies for bacterial pneumonia-induced injury.

NIH Research Projects · FY 2026 · 2024-06

Project Summary/Abstract: Pharmacologic treatment of acute ischemic stroke (AIS) is very challenging, largely due to the difficulty of rapidly and specifically delivering drugs to the ischemic region of the brain. Targeted drug delivery using affinity moieties (e.g., antibodies) can increase the local concentration of drugs, thereby improving the therapeutic index. But targeting to blood brain barrier (BBB) epitopes such as transferrin and insulin receptors achieves poor delivery with intravenous administration (e.g., with intravascular targeted-nanocarriers, T-NCs) and is not selective to the injured region of the brain. New targeting/delivery approaches are needed for AIS. We and others have demonstrated that Vascular Cellular Adhesion Molecule (VCAM) is highly overexpressed in the endothelial cells of the penumbra (brain regions at risk of death). Here, we will exploit this for targeted drug delivery with T-NC. We have demonstrated how VCAM T-NC provides unprecedented enhancement of drug delivery to the brain after IV administration (>20 times higher vs. precedent approaches), specifically to the injured brain. In our pilot studies in an animal model of AIS, we observed rapid and specific uptake of CAM-targeted mAbs and T-NCs encapsulating mRNA encoding for interleukin 10/IL-10 (anti- inflammatory cytokine), providing targeted treatment that reduced stroke volume by >60% and improved survival from 60% to 100%. Central hypothesis: VCAM-directed T-NCs can restore BBB function by delivering therapeutic RNAs to endothelial cells in the ischemic penumbra and core, limiting sustained inflammation and brain damage after AIS. We will deliver IL-10 mRNA and siRNA against ICAM with T-NCs, to promote a local anti- inflammatory environment that will restore BBB homeostasis. Our T-NCs: i) Will target RNAs to the BBB to decrease BBB permeability to plasma proteins and reduce leukocyte infiltration; ii) Will deliver RNAs with complementary temporal effects to act at different phases of the disease – IL-10 will reduce inflammation in the acute phase of AIS, and ICAM siRNA will protect during the subacute one. In the current proposal, we suggest: Aim 1: Vascular immunophenotype and nanocarrier delivery in AIS. Aim 2: CAM T-NC to enhance the BBB after AIS. Our deliverables are: i) generate a spatiotemporal map for the expression of CAMs in the AIS of mice and its vascular accessibility to deliver T-NC, ii) Independently of the therapeutic efficacy of our approach will validate our central hypothesis as we are going to quantify: i) expression of the therapeutic mRNA, ii) silencing of ICAM in the BBB, and iii) cellular infiltration. We will also identify in an unbiased way different pathways affected by ischemic stroke and how these pathways are altered after the treatment with anti-inflammatory molecules. This will help to identify new targets for treatment. In addition, we will identify the possible toxicities associated with this therapeutic approach. These results will be key for further investigations in large animal models.

NIH Research Projects · FY 2026 · 2024-04

Voice disorders occur in 6–17% of children, resulting in dysphonia (i.e., altered vocal quality) which has significant negative health, social, emotional, and educational consequences if left untreated. The most common cause of dysphonia in children is vocal fold nodules (VFN), callus-like growths on the vocal folds. Although voice therapy is the preferred treatment, over one-third of children do not show clinically meaningful improvements from therapy. Two critical issues contribute to this lack of impactful therapeutic changes: 1) children are not mini- adults, and 2) dysphonia reduction may not be a salient and motivating goal for all children. To maximize treatment effectiveness for the dynamic pediatric system, we propose shifting from a narrow focus on dysphonia to a broader focus on improving speech intelligibility. This proposal will examine previously collected speech samples from children with VFN between 3 – 9 years, a critical developmental period. Speech samples will be compiled from clinical databases from three top pediatric voice centers, Children’s Hospital of Philadelphia, Boston Children’s Hospital, and Cincinnati Children’s Hospital Medical Center. Aim 1 will define the relationship between dysphonia and measures of intelligibility (e.g., speech intelligibility, speech naturalness). As intelligible speech production requires coordination of the vocal and articulatory systems, we hypothesize that the significant developmental changes in these systems will impact the relationship between dysphonia and intelligibility. Aim 2 will determine the extent that acoustic measures of the developing vocal system, developing articulatory system, and dysphonia severity contribute to the relationship between dysphonia and intelligibility. The acoustic measures with the largest contribution to the relationship between dysphonia and intelligibility will be examined in a subsequent R01 grant to determine which therapy tasks elicit the largest change in these measures. These therapy tasks will then be examined in a randomized controlled trial to test their efficacy and effectiveness in improving intelligibility in children with VFN. This research program is designed to address multiple priority areas of the NIDCD, with an overall emphasis on improving human communication (Priority Area 4). The current grant focuses on understanding diseases and disorders (Priority Area 2), with outcomes indicating which acoustic measures are the ideal method of predicting intelligibility in this population. This R21 lays the groundwork for the subsequent R01, a randomized clinical trial designed to test the effectiveness of therapy tasks at different developmental stages. This research program will therefore optimize treatment for children with VFN (Priority Area 3), providing information on developmentally appropriate therapy tasks for this underserved and vulnerable population. The research will be completed at Temple University (R1 university), which maintains a robust research environment with exceptional dedication and support to early-career faculty. Outcomes from this proposal will lay the foundation for a paradigm shift in how we conceptualize and treat children with voice disorders, an understudied and underserved clinical population.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Diabetes mellitus is a common chronic metabolic disease imposing great social and economic burden, and is considered as one of the major health-threats in USA and worldwide. The morbidity of critical limb ischemia (CLI) in diabetic patients is extremely high (up to 76% in some studies). Therefore, understanding the mechanisms of CLI pathogenesis in diabetes is critical for developing novel therapeutic strategies. We and others have reported that enhancing endothelial cell (EC) and endothelial progenitor cell function improves ischemic tissue repair. Recently, there is growing evidence indicating that diabetes severely impairs angiogenic property of EC/EPC that may directly limit CLI repair. The overall goal of this proposal is to elucidate the role of muscle-specific miR-499 in diabetes-impaired EC/EPC function. The premise of our proposed studies is based on our preliminary studies showing that expression of miR-499, a muscle-specific miR, was enhanced in EC/EPCs from db/db mice. Our central hypothesis is that diabetes e nhanced miR-499 impairs EC/EPCs function and ischemic limb repair via skeletal muscle-derived exosomal delivery of miR-499 to EC/EPCs. We propose to conduct complementary experiments organized under the following 3 aims: 1. Determine the role of miR-499 in diabetes-induced EC and EPC dysfunction; 2. Demonstrate the role of muscle-derived exosomes in miR-499-induced EC dysfunction in diabetes; 3. Elucidate the molecular signaling downstream of miR-499 involved in diabetes-impaired angiogenic property of EC. Our findings will provide fundamental insights into development of novel strategies for therapeutics of CLI in diabetic patients. 1

NIH Research Projects · FY 2025 · 2024-04

SUMMARY Understanding the mechanisms underlying GPCR signaling is crucial in order to fully comprehend their role in physiology and pathophysiology. In addition to canonical second messengers (cAMP, cGMP and IP3) and β- arrestin signaling, small GTPase proteins, such as Rho GTPases are largely involved in GPCR-mediated signal transduction. Guanine nucleotide exchange factors (GEFs) convert Rho GTPases from an inactive (GDP-bound) state to an active state (GTP-bound). Rho-GEFs can be activated by Gq, G12/13 and Gs proteins. However, currently there is no evidence that Gi/0-WT can directly activate RhoGEFs. PDZ domains are structural protein domains that recognize simple linear amino acid motifs often at the protein C-terminal (C-motif). RhoA, activated by PDZ-RhoGEFs, has important signaling roles, by activating phospholipase D (PLD) and transcription factors. Kappa opioid receptor (KOR), an abundantly expressed GPCR that mainly couples to Gi/o, has a KPV C-motif (last three amino acids) that binds PDZ class III proteins, including PDZ-RhoGEF. Our central hypothesis is that KOR activation, in addition to engaging cAMP inhibition and β-arrestin pathways, initiates an additional signaling mechanism downstream to PDZ-RhoGEF leading to activation of RhoA and subsequent activation of PLD. Activation of PLD generates two distinct second messengers, phosphatidic acid, which activates the mTOR pathway, and choline, which activates Sigma1 receptors. In addition to in vitro studies, this pathway that plays an essential role in KOR signaling will be studied in vivo on antipruritic effects of KOR agonists. Antipruritic effects of KOR agonists are well established and as of today, there is one FDA-approved KOR agonist (difelikefalin) for the treatment of chronic itch of chronic kidney diseases. We will use a multidisciplinary approach, combining state-of-the-art molecular and pharmacological approaches for a comprehensive investigation of the signaling pathways elicited by activation of C-motifs of KOR. Receptor and PDZ-RhoGEF mutations, measurements of second messenger levels (cAMP, choline and phosphatidic acid) as well as live imaging of PLD activation will be used. Acute itch model of scratching will be used for in vivo studies. We provide solid preliminary results supporting the feasibility of the project and the ability of our team to complete the work proposed. The project has two aims: Aim 1. Investigate the role of PDZ-binding domain in KOR-induced signaling; experiments are designed to characterize intracellular cascades activated by KOR-motifs (KOR-PDZ binding domain). Aim 2. Investigate the role of PDZ-RhoGEF/RhoA pathway in antipruritic effects of KOR agonists. RoA and sigma 1 receptor antagonists will be tested on scratch inhibiting effects of three different KOR agonists (U50,488 as prototype, nalbuphine, and difelikefalin as peripherally restricted). The successful completion of this project will increase the current knowledge of GPCR signaling and will serve as a basis for further development of ligands selectively targeting this pathway and drug development in the itch field.

NIH Research Projects · FY 2026 · 2024-04

Pediatric obesity is an epidemic with striking disparities by socioeconomic status (SES) and race/ethnicity. Thus, there is a need to identify novel approaches for obesity prevention, particularly for those at greatest risk. Primary care may be an ideal setting for obesity prevention given the high percentage of children who attend annual well child visits and recommendations that lifestyle behaviors be addressed in primary care with all children who do not yet have obesity. However, there is limited support for effectiveness of obesity prevention for school-aged children in primary care. Thus, identification of effective and efficient approaches that are well suited for translation are imperative. One such efficient approach may be enhancing children’s sleep. A good night’s sleep in childhood is a well-known correlate of healthier weight status. Adult experimental studies demonstrate that increasing sleep leads to positive changes in eating behaviors and weight. However, few interventions to enhance sleep duration have been conducted with school-aged children and we are unaware of any in primary care, which is striking given the large percentage of children with insufficient sleep and accumulating evidence supporting the importance of sleep for weight regulation. Our own randomized controlled trials with school-aged children provide compelling preliminary evidence that a brief behavioral intervention can result in clinically meaningful improvements in children’s sleep duration and that enhancing sleep can lead to positive changes in eating and activity behaviors and weight status. However, our studies have been limited to academic settings. Thus, the purpose of the proposed study is to determine (from a RE-AIM – reach effectiveness, adoption, implementation, maintenance – framework) feasibility, acceptability, and preliminary effectiveness of our sleep intervention for obesity prevention in primary care. The intervention’s brevity, flexibility in delivery mode (e.g., via zoom/phone), and consistent signal for efficacy enhances translation potential. Further, we propose enhancing reimbursement potential by modeling our intervention delivery after an existing reimbursable model of care for another chronic health condition (asthma) in which pediatricians refer for disease management support by a registered nurse (RN). Fifty short sleeping (< 9 hours/night) children 6-11 years old who primarily identify as African American/Black and from under-resourced communities will be enrolled. Children will be randomly assigned to either: 1) optimize sleep primary care (OSPC; our 4-session behavioral intervention to enhance sleep duration delivered by a RN in primary care) or 2) enhanced usual care (EUC; usual care plus sleep education delivered at the same timepoints as OSPC). At baseline, end of treatment (2 months), and 6 months, the following will be measured: feasibility/acceptability metrics, sleep duration (actigraphy), eating behaviors (24-hour dietary recalls), physical activity (accelerometry), and anthropometrics (measured height and weight).

NIH Research Projects · FY 2026 · 2024-03

Abstract This molecular most is a proposal to use fetal-derived exosomes isolated non-invasively from maternal blood as a source of biomarkers that predict whether an at-risk fetus will be born with fetal alcohol syndrome (FAS), the involved end of the fetal alcohol spectrum disorders ( FASD), or one of the partial forms . Prenatal alcohol exposure is the most common cause of intellectual disability in the US. Other features of FAS include small eyes, face, and brain, as well as neurobehavioral deficits. These syndromes are not readily detected early in pregnancy with available imaging techniques, but early diagnosis could facilitate development of therapeutic interventions. We previously studied banked human fetal brains and eyes at 9-22 weeks' gestation paired with maternal blood samples from pregnancies affected by maternal alcohol use (assessed by self-report) compared with non- exposed controls and demonstrated high correlations between altered expression of several protein, RNA and apoptotic signaling molecular markers and morphometric abnormalities consistent with FAS, such as reduced eye diameter and brain size. The molecular abnormalities were detectable in fetal brain-derived exosomes (FB- E) isolated from the maternal blood. Now, we propose a follow-up clinical study to determine whether these biomarkers will predict the postnatal emergence of FAS/FASD. In collaboration with the Department of OB/GYN and the Section of Neonatology at Temple University Hospital, FB-E will be isolated from maternal blood and analyzed for protein, RNA, and apoptotic markers throughout the pregnancy, and correlated with morphological observations to detect features of FAS/D (small eye globes, small head circumference, longer upper lip, absent philtrum, etc.) in newborn infants, with follow-up pediatric exams for 2 years. We will compare findings in 150 EtOH-exposed pregnancies with 75 unexposed pregnancies, controlling for infant sex, maternal obesity, age, race/socioeconomic status, and tobacco use, and excluding cases in which the mother used other substances of abuse, to determine which biomarkers are most predictive of FAS/FASD in at risk newborns. Pregnant women will fill out a standard survey about their use of alcohol, and cell-type specific (oligodendrocyte, neuronal, microglial, astroglial, synaptic) FB-Es will be isolated from their blood. The molecular contents of these exosomes will be assayed for biomarkers, particularly ones associated with dysmyelination, e.g., MBP, reductions of which showed the most consistent correlation with alcohol-associated reductions in eye diameter in our previous studies. The findings will be correlated with morphological abnormalities characteristic for FAS/D in neonates. If it works in FAS/D, this approach might be useful to predict other disorders of neurological development.

NIH Research Projects · FY 2025 · 2024-03

Abstract: With increased use of fentanyl or fentanyl-analog compounds in the United States, the opioid epidemic has evolved into a `fentanyl epidemic.' Overdose deaths involving these extremely potent mu-opioid receptor (MOR) agonists have increased ~30-40-fold in the last 20 years. Moreover, in 2021, drug overdose deaths in the United States topped 100,000 for the first time ever and more than half of all deaths involved fentanyl or fentanyl analogs. Yet, the brain neuroadaptations induced by fentanyl and fentanyl analog exposure remain poorly described. Additionally, it is not known if the high amount of fentanyl-associated deaths are due to fentanyl being `more addictive' than other opioids, or if fentanyl induces a unique set of neuroadaptations associated with increased opioid seeking. The limited literature describing molecular alterations following fentanyl exposure in comparison with other well-studied opioids such as morphine indicates divergency amongst MOR agonists for neuroadaptations involving microRNA (miRNA) regulation and gene expression. miRNAs inhibit protein translation to modulate gene expression and have recently emerged as critical regulators of drug seeking for many drug classes. miRNAs have additional utility as biomarkers because miRNAs are present in exosomes found in serum of peripheral blood samples. Investigation into the relationship between drug exposure and regulation of brain miRNAs that can also be detected in the periphery can be accomplished easily with rodent models of self-administration but has yet to be done. In this exploratory project, we will use rat opioid self-administration to: 1) examine the acute and long-lasting effects of fentanyl or the ultra-potent analog furanylfentanyl on exosomal blood serum and orbitofrontal cortex (OFC) brain microRNA (miRNA) profiles; 2) compare the fentanyl-induced miRNA profiles to the less potent opioid heroin; 3) correlate fentanyl-induced blood and brain miRNA profiles to opioid seeking behavior at multiple timepoints; and 4) directly compare fentanyl-, furanylfentanyl- and heroin-induced drug seeking behavior at response- equivalent dosages to identify agonist-specific differences in long-lasting drug seeking behavior. This project will provide critical insight into both the immediate and long-lasting molecular consequences of fentanyl and furanylfentanyl exposure on brain neurochemistry and identify miRNA-mediated pathways associated with opioid seeking. Therefore, this study provides an essential opportunity to understand the relationship between blood and brain miRNA expression as an indication of opioid craving. Studies of blood miRNA profiles associated with drug seeking behavior have translational potential, as biomarkers indicative of recovery from substance use or relapse may help to inform patient care or treatment responsiveness.

NIH Research Projects · FY 2026 · 2024-03

SUMMARY Between 2000, when the first version of Hypothesis Testing Using Phylogenies (HyPhy) was released, and 2022, the number of bases in GenBank increased ~250 fold, the number of sequenced genomes from a handful to >3,5003, and the number of PubMed papers studying molecular evolution ~12 fold. Data generation has ceased to be the bottleneck for biological and biomedical discoveries, and the new bottleneck is methods, tools, and software for data analysis and interpretation. Comparative evolutionary analyses remain an essential and powerful method for extracting meaning and insight from ever-expanding genomic sequence data. But, the lack of emphasis and incentive structure for developing, maintaining, benchmarking, and improving software and analysis tools, despite their essential and critical role in modern biology, biotechnology, and medicine remains a major concern. Analytical, infrastructure, and incentive challenges exposed by the genomic data deluge during the COVID-19 pandemic were aptly summarized. Over the last quarter century HyPhy has established itself as a useful, popular, and durable platform for studying diverse evolutionary processes, such as natural selection and recombination, across different taxonomic scales. Datamonkey, a web application providing free access to “one-click” popular HyPhy analyses, has seen increasing use by researchers worldwide over the last two decades. Through continued methodological innovation, improvements in performance and scalability, accessible web services, focus on data visualization, and user support, HyPhy developers were able to sustain and increase the reach and impact of the program. This proposal seeks to improve the software, enhance biological realism, relevance, accessibility, and interpretability of the methods, design novel approaches to address outstanding problems in evolutionary data analysis, and further lower access barriers to evolutionary comparative analyses through integration with the Galaxy ecosystem.