University Of Pennsylvania

NIH Research Projects · FY 2026 · 2025-08

1 PROJECT SUMMARY: 2 Cardiac Allograft Vasculopathy (CAV) is the leading cause of transplanted heart failure and loss. Despite being 3 a major cause of morbidity for heart transplant (HT) recipients, little progress has been made in tailoring CAV 4 surveillance and mitigation strategies to individual patient risk. All guideline-endorsed methods for CAV detection 5 rely on repurposing techniques initially developed for evaluating native coronary artery disease. As such, these 6 techniques focus on estimating the flow of blood through the large, epicardial coronary arteries. Despite the 7 large-vessel focus of established CAV screening, the pathobiology of CAV also involves significant microvascular 8 inflammation, with distinct histologic changes which precede overt, large-vessel manifestations. Currently, there 9 are no objective means for measuring these microscopic changes, forcing clinicians to rely on tests which only 10 detect CAV after it has progressed to macroscopic stages. We hypothesize that the application of digital tissue 11 analysis methods in CAV will lead to both a clinically viable risk-assessment tool, and to the discovery of novel 12 mechanistic biology. Human tissue samples contain a wealth of information which is underutilized in conventional 13 clinical and scientific workflows. However, advanced digital technologies capable of extracting and quantifying 14 the spatial information contained within residual tissues are poised to change this. Computer-aided image anal- 15 ysis of digital pathology (DP) slides can extract novel ‘morphologic biomarkers’, quantifying the sizes, patterns, 16 and spatial relationships which comprise the cardiac microarchitecture. Combining DP morphologic analysis with 17 new, spatial-molecular profiling techniques adds additional insights, enabling deep, mechanistic interrogations. 18 Our team is a leader in cardiac digital pathology (DP) analysis, having developed numerous first-in-heart pipe- 19 lines for generating clinically important diagnoses and risk-assessments. We are also leaders in spatial-molec- 20 ular profiling of human cardiac tissues, with important contributions in both HT and non-HT diseases. As de- 21 scribed in a recent publication in Circulation, we have developed the ‘integrated-CAV Prediction’ (iCAV-Pr) sys- 22 tem. iCAV-Pr uses clinical data and residual tissue from routine post-HT biopsies to predict which patients will 23 go on to develop CAV years before overt disease onset. To fill unmet needs in heart transplant care, we propose 24 a multicenter study aimed at validating iCAV-Pr predictive performance, at establishing it’s potential role in clin- 25 ical practice, and at defining the biology that underlies it’s predictions. In Aim 1, we will enhance the iCAV-Pr via 26 extraction of new morphologic features, followed by performance validation in a diverse, multinational, retrospec- 27 tive cohort. In Aim 2, we will assess the efficacy of iCAV-Pr vs. intravascular ultrasound, the most sensitive 28 existing method for detecting early changes in CAV. And, in Aim 3, we will perform high-plex digital spatial 29 proteomic profiling on serially collected EMB pathology slides to deeply explore the mechanisms underlying CAV 30 development. Successful completion of these aims could open a new frontier in personalized post-transplant 31 care, enabling providers to tailor both CAV screening and treatment strategies to individual patient risk.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY/ABSTRACT: This application requests funding to rigorously study the effectiveness and implementation of Hospital-to-Housing, a large-scale intervention that provides medically supported transitional housing for people experiencing homelessness with opioid use disorder (OUD) following hospitalization. Homelessness is associated with poor health outcomes, many of which stem from consequences of substance use. In addition, homelessness complicates access to medical care, including substance use treatment, and increases acute care utilization. Medical hospitalization represents a critical, reachable moment for patients with OUD to initiate treatment with highly effective medications such as methadone and buprenorphine. However, patients hospitalized with complications from OUD have complex ongoing needs for medical care, substance use treatment, and mental health conditions after hospital discharge that are challenging to manage without stable housing. Hospital-to-Housing (H2H) is community-academic partnership between Project HOME, a nationally recognized housing organization, and three large Philadelphia health systems to transform care for people with OUD, medical complexity, and unsheltered homelessness. The H2H model includes several evidence-based components, including post-discharge transitional support, respite housing with integrated medical care and integrated OUD treatment, case management support, and transition to permanent supportive housing, all in a low-barrier model that does not require abstinence for continued housing and treatment. The program, funded through a philanthropic investment, does not include a formal evaluation, creating a time- sensitive opportunity to advance our understanding of strategies to address social needs of people experiencing homelessness, OUD, and medical complexity. The aims of this proposal are to: 1) Estimate the effect of H2H on engagement in outpatient substance use treatment and acute care use (primary outcomes), housing stability, engagement in primary care, and overdose and all-cause mortality (secondary outcomes); 2) Explore active components of H2H and identify barriers and facilitators to implementation and scale; and 3) Conduct a cost and cost-effectiveness analysis of the H2H model. To complete the aims, we will leverage partnerships between Project HOME, participating health systems, and Philadelphia city agencies to access administrative data and use target trial emulation and quasi-experimental techniques to compare individuals enrolled in H2H during hospitalization with a cohort of matched controls. The proposed work uses innovative methods to estimate real- world effectiveness of a medically supported housing model, implemented at scale in a major city at the epicenter of the overdose crisis and without additional research-funded resources. Such models are critically important given the rise in OUD-related complications, such as infections and wounds, that are poorly managed in our current treatment system. Successful completion of this study will provide rigorous evidence about the effectiveness of H2H and critical information to inform implementation and scaling of future models.

NIH Research Projects · FY 2025 · 2025-08

In the US, there are significant, unacceptable disparities between Black and non-Black women in cesarean delivery rates, as well as maternal and neonatal morbidity. Standardization of labor induction is a promising intervention to reduce such racial disparities in obstetric outcomes. In a completed prospective cohort study, our group used a pre- and post-implementation mixed- methods analysis to determine effectiveness of our induction protocol on overall obstetric outcomes, as well as racial disparities, while simultaneously evaluating implementation outcomes across two sites. To encourage implementation of the induction protocol, we harnessed several implementation strategies, including unit-level audit and feedback reports distributed by site. As a part of this work, we performed qualitative interviews with site clinicians to determine protocol acceptability, as well as assess our implementation endeavors. A critical theme emerged: clinicians felt that these general, unit-level audit and feedback reports demonstrating disparate outcomes could not represent their individual care; they needed individual-level feedback to feel responsible. The central hypothesis of this proposal is that a Disparities Dashboard, detailing clinician- level utilization of evidence-based practices, as well as clinical outcomes, by patient demographics, has the potential to drive clinician behavior change. We plan to test this hypothesis in a 2-aim approach. First, we propose a three-phase mixed-methods study that includes iterative feedback from clinicians and patients with the purpose of developing an acceptable Disparities in Labor Outcomes Dashboard. Second, we plan to compare 6 months where both sites will continue to use unit-level audit and feedback as the primary implementation strategy, to 6 months where Site #1 will continue to use unit-level audit and feedback, but clinician-level Disparities Dashboards will be implemented at Site #2. We will evaluate the Dashboard’s impact on intervention fidelity (adherence to the labor induction protocol) as well as clinical outcomes potentially impacted by increases in these evidence-based practices (cesarean delivery, maternal morbidity) in a difference-in-differences study design. Creation and evaluation of Disparities Dashboards as an implementation strategy to reduce disparities in obstetrics is an important clinical question yet to be studied in the literature. Dr. Hamm is an R01 and U24-funded maternal fetal medicine physician trained in clinical epidemiology with an established interest in implementation research. This proposal will leverage collaboration with senior investigators (Drs. Srinivas, Ashcraft, and Howell) and Penn’s programs in obstetrics, implementation science, health disparities, and qualitative methods. The results of this R21 will be invaluable to supporting a planned R01-level application for a stepped-wedge hybrid effectiveness implementation trial evaluating implementation of the standardized induction protocol at diverse sites, using the Disparities Dashboard as a critical implementation strategy for success.

NIH Research Projects · FY 2026 · 2025-08

Summary The goal of this proposal is to test a novel role for actin during mitosis in the preimplantation mouse embryo. Following fertilization, the embryo starts to undergo mitotic divisions and must accurately propagate its genetic material in order to avoid aneuploidy. However, early mammalian embryos lack efficient spindle assembly mechanisms and it remains unclear how proper chromosome segregation is achieved during early development. Using advanced imaging, we discovered actin cables inside the cell nucleus in the early embryo. Upon mitosis, these cables capture chromosomes and undergo contraction. This initiates a chromosome congression-like process before the mitotic spindle completes its assembly, which is disrupted by manipulation of F-actin, but not of microtubules. Moreover, when the spindle is assembled, we found another actin network that encloses the spindle and regulates its growth and position, which likely helps to ensure proper cell division patterns. Our data support a new paradigm for the regulation of mitosis in early development whereby nuclear actin cables and peri-spindle actin regulate key steps of mitosis, typically carried out by the mitotic spindle in centrosomal cells. Thus, we propose to dissect the mechanisms by which these actin networks form, how they are regulated, their importance for ensuring mitotic fidelity, and their developmental regulation. Aim 1 will determine how nuclear actin cables form, capture chromosomes, and initiate their congression. For this, we will combine high-resolution live-imaging of actin, chromosome, and microtubule dynamics with biophysical and molecular manipulations in the intact-developing mouse embryo. Aim 2 will use similar approaches to mechanistically dissect how the peri-spindle actin network forms, and how it regulates spindle growth and position to ensure proper chromosome segregation and cell division patterns. In addition to testing a new actin-dependent model for the regulation of mitotic fidelity during early development, the project may eventually help to reduce mitotic errors in IVF embryos, a significant barrier in reproductive medicine.

NIH Research Projects · FY 2025 · 2025-08

Summary Antibody-based approaches to HIV cure or ART-free remission hold promise. Several antibody-based therapeutic HIV vaccine trials are currently underway, leveraging germline-targeting approaches originally developed for HIV prevention, which ignore the existing autologous neutralizing antibody (anAb) responses already present in people with HIV (PWH). Here, we propose an alternative strategy of personalized therapeutic immunization to expand and enhance PWH's own anAb responses with the goal of eliciting antibody-mediated viral control and ART-free remission. We hypothesize that a personalized anAb-boosting vaccine administered after early ART initiation can boost anAb breadth and potency sufficiently to neutralize the replication-competent viral reservoir and elicit ART-free virus control. Recent findings from our group and others support this rationale: 1) Early ART limits reservoir diversity while enabling ongoing anAb evolution; 2) AnAb responses restrict viral rebound after treatment interruption, and correlate with delayed time to rebound; 3) Our TF SHIV-infected nonhuman primate model recapitulates HIV immunopathogenesis, persistence, and the kinetics and potency of anAb responses; 4) New insights into antibody development via vaccination and SHIV infection enables rational immunogen design; 5) Novel mRNA-lipid nanoparticle delivery and Env engineering technologies allow rapid production of personalized stabilized Env immunogens. We propose a coordinated 3- arm rhesus macaque experiment to test whether individualized mRNA-LNP delivered Env immunogens can enhance baseline anAb responses and elicit durable ART-free virus control compared to off-the-shelf germline targeting immunogens or ART alone. We will test a personalized two Env immunogen strategy, based on the unique Env sequence diversity present at early ART initiation in each animal, aiming to engage baseline responses and then broaden them to cover the diversity of viruses in each animal. We will test these vaccination regimens in macaques infected with a validated, barcoded SHIV and early ART treatment. Nine animals in each arm will be (A) immunized with two doses of mRNA-LNP delivered stabilized Env representing the most frequent natural month 4 Env variant in each animal (Env.con), then two doses of a distinct Env designed to more fully cover the diversity of sampled month 4 virus (Env.div), (B) immunized with two mRNA-LNP-delivered doses of an off-the-shelf germline targeting immunogen (BG505.GT1.1), then two doses of the native BG505 Env, or (C) ART alone. Primary study outcomes will be impact of vaccination on post-ART virus control and enhancement of anAb responses. We will also conduct mechanistic studies to characterize virus rebound and identify blood and tissue correlates of post-ART virus control. Results will advance the HIV cure field by defining the potential of HIV-specific humoral responses and the capacity of novel immunogens to grow them. If successful, this approach could be central to a combination cure strategy by pairing it with cellular vaccines or other immunotherapies, then iteratively extending it to PWH on chronic ART, and rapidly translating it into human trials.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT The gut microbiome plays a crucial role in health, and its disruption by antibiotics can lead to decreased microbial diversity, loss of essential metabolic functions, and increased susceptibility to infections with pathogens such as Clostridioides difficile. The mechanisms and factors influencing the recovery of the microbiome from antibiotics remain inadequately understood. Notably, the potential impact of companion animals on the human microbiome is often overlooked, even though recent evidence suggests that pets can modulate the human microbiome and protect against colonization and recurrence of infection with C. difficile. To investigate these dynamics, the REMBRANDT study (REcovery of the MicroBiome fRom ANtibiotics for Dental implanTs) was initiated as a longitudinal cohort study examining microbiome disruption and recovery in pet owners and non-pet owners receiving antibiotic prophylaxis for dental surgeries. Stool and saliva samples are collected from participants, and if applicable, their pets, before, during, and after the antibiotic regimen, alongside extensive meta-data. Currently, metagenomic 16S rRNA sequencing is performed on all samples, which provides genus-level taxonomic data. Preliminary analyses appear to confirm that pet ownership can influence the post-antibiotic trajectory of the microbiome. Here, we propose to sequence REMBRANDT samples more deeply using shotgun metagenomic sequencing (SMS) to achieve higher-resolution taxonomic profiling of the microbiome and its associated gene families and metabolic pathways. This enhanced sequencing approach will allow for a comprehensive characterization of the taxonomic and functional disruption and recovery of the microbiome and a more sensitive detection of patient and household-level characteristics that affect recovery. Additionally, this approach will provide insight into the post-antibiotic collection of antimicrobial resistance genes in the gut (the resistome) and the factors that govern the acquisition and persistence of antimicrobial resistance genes within the host and their potential dissemination to pets. Findings from this research will be used to inform public health strategies and interventions aimed at enhancing microbiome recovery and limiting the spread of antimicrobial resistance.

NIH Research Projects · FY 2025 · 2025-08

Extracellular vesicles (EVs) have gained significant attention for their diagnostic and therapeutic potential due to their unique protein and RNA cargo, which play key roles in cancers and other diseases. However, despite extensive research and investment, the clinical translation of EV biomarkers remains limited. A major challenge is that EVs from diseased tissues often have molecular similarities to those from healthy cells, with only subtle differences in their cargo. The specific packaging of molecular cargo in individual vesicles is believed to be crucial to their function, making bulk analysis methods insufficient. Single EV-level analysis is essential. Current approaches, relying on low-throughput, high-resolution imaging, are hindered by the vast number of EVs in clinical samples like blood (~1010 EVs/mL). To overcome this, we propose Agarose Bead-based Digital Single Molecule-Single EV Sorting (BDEVS), a high- throughput microfluidic platform for ultra-sensitive profiling of individual EVs in plasma. Unlike conventional methods, BDEVS provides single-molecule sensitivity and multiplexing (>5 targets) without compromising throughput, analyzing millions of EVs per minute. The platform uses rolling circle amplification (RCA) of cleaved EV surface proteins, which are amplified in microscale agar droplets, followed by flow cytometry-based readout and sorting. This approach addresses previous limitations such as steric hindrance, non-specific binding, and limited protein quantification on EVs.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Skin-penetrating helminths affect 1.5 billion people worldwide and can cause clinically significant itch. However, whether itch-inducing neurons regulate skin anti-helminth immunity is unknown. Cutaneous infections with the helminth Schistosoma mansoni cause mild itch and little skin irritation that result in parasite dissemination. My recently published studies demonstrate that S. mansoni suppresses itch evoked by neurons bearing the Mas- related G protein-coupled receptor (Mrgpr) A3. However, the molecular mechanisms by which S. mansoni inhibits MrgprA3 neurons are undefined. My published studies also showed that activation of MrgprA3 neurons induced IL-17-mediated inflammation, keratinocyte thickening, and skin resistance to S. mansoni dependent on antigen- presenting cells. However, the effector mechanisms by which MrgprA3 neurons induce skin inflammation and anti-parasitic immunity are obscure. Two main subsets of itch-transmitting neurons are defined by MrgprA3 (NP2) and MrgprD (NP1) in mice, but these two neuronal subsets express remarkably distinct transcriptional programs and have unique functions. In contrast to the pro-inflammatory properties of MrgprA3 neurons, my preliminary data show that MrgprD neuron stimulation limits IL-17-driven skin inflammation and promotes secretion of the anti-inflammatory cytokine IL-10, which is well-known to promote S. mansoni systemic dissemination. Based on these studies, this K99/R00 proposal is designed to test the central hypothesis that S. mansoni inhibits MrgprA3 neurons that initiate IL-17-driven keratinocyte responses but preferentially stimulates MrgprD neurons that suppress skin immunity by inducing IL-10 secretion. This hypothesis will be tested in three specific but complementary aims. Aim 1 (K99) will determine how S. mansoni blocks the activation and itch evoked by MrgprA3 neurons. Aim 2 (K99/R00) will test if MrgprA3 neurons protect against S. mansoni by eliciting IL-17- dependent keratinocyte proliferation and maturation. The R00 independent phase in Aim 3 will leverage the expertise acquired in Aim 1 and 2 to define if S. mansoni directly activates MrgprD neurons to promote IL-10- dependent immunosuppression that facilitates its dissemination. The studies of this K99/R00 proposal may inform new therapies against chronic itch and skin helminth infections. The principal investigator (PI), Dr. Juan Inclan-Rico, will learn the skills and techniques necessary to accomplish the proposed research under the guidance of his mentoring team, led by Drs. Wenqin Luo and De’Broski Herbert, who have pioneering expertise in somatosensation and immunoparasitology. Collectively, the mentoring committee has a proven record of transitioning postdoctoral fellows into independent investigators. Dr. Inclan-Rico will also acquire the leadership, management, mentorship, and grantsmanship skillset necessary to run his own laboratory. The PI’s strong enthusiasm, publication record, and commitment to science, combined with the training obtained during the K99 mentored phase, will define a clear path to establish his independent research program that defines the contributions of itch-inducing neurons to skin immunity during infectious and non-infectious conditions.

NIH Research Projects · FY 2025 · 2025-08

Despite continuous efforts to eradicate tuberculosis (TB), Mycobacterium tuberculosis (Mtb) is still the leading infectious agent worldwide. Most people heavily exposed to Mtb become infected and develop latent Mtb infection (LTBI). Importantly, Dr. Hong’s collaborators identified ~7% of individuals who are heavily and repeatedly exposed to Mtb resist developing LTBI (resister or “RSTR”) in a household contact (HHC) study in Uganda. However, clinical and epidemiological studies were not able to differentiate these clinical phenotypes (RSTR vs. LTBI). To discover genetic determinants of this resistance phenotype, Dr. Hong mapped expression quantitative trait loci (eQTLs) associated with gene expression in Mtb-infected, but not uninfected monocytes from the same patient cohort. This genetic approach revealed that 29 eQTLs were significantly associated with the expression of 16 genes in response to Mtb infection. Despite the discovery of Mtb-specific genetic signatures, the functional importance of these eQTLs and their target genes in macrophage responses to Mtb infection deserves to be further investigated. In the current proposal, the primary goal is to discover the global human genetic regulators (genes and variants) of Mtb-induced macrophage antimicrobial pathways. Dr. Hong hypothesizes that Mtb-dependent eQTL monocyte response genes (termed “Mtb eQTL gene”) and variants are global regulators of macrophage responses to Mtb and mediate critical proinflammatory and antimicrobial responses. Alternatively, these genes may be hijacked by Mtb to favor its survival. In Aim 1, she will determine the mechanisms of how Mtb eQTL genes regulate macrophage antimicrobial responses to Mtb and other pathogens. In Aim 2, she will determine the allele-specific antimicrobial macrophage mechanisms of Mtb eQTLs and their association with clinical outcomes. Significant findings from this project will enrich our understanding of how Mtb eQTL genes and variants modulate macrophage responses to Mtb, directing towards understanding why individuals have different susceptibility to Mtb infection and TB disease as well as whether these insights can lead to novel treatment or diagnostic strategies. Dr. Hong, Assistant Professor at the University of Pennsylvania School of Nursing (primary) and the Perelman School of Medicine (secondary), is a clinician scientist with a Ph.D. in TB research and epidemiology. Dr. Hong is seeking a K08 Mentored Clinical Scientist Research Career Development Award to acquire advanced training and mentorship for further studying the influence of genetic determinants on resistance and susceptibility to Mtb infection and TB disease. Since her research focus has shifted from clinical epidemiology to immunogenetics in TB, this K08 training award will provide her with the necessary protected time to fill the gaps in her research skills. By implementing the studies described in this proposal under the guidance of expert mentors, Dr. Hong will be well-positioned as an independent investigator, characterizing novel mechanisms of TB susceptibility with expertise in TB immunogenetics.

NIH Research Projects · FY 2025 · 2025-08

R50 Abstract – Reiss Gastrointestinal maligancies remain lethal and difficult-to-treat, with poor outcomes for many patients. In spite of this, we are in a moment of a scientific and clinical revolution with the development of biomarker-driven therapies for selected patients. With these advances come multiple new questions for the field, many of which the NCTN infrastructure is optimally suited to address. To date, I have demonstrated a strong commitment to clinical and translational research for populations of patients with rare subsets or biomarkers, including via the development and implementation of NCI-trials at my own institution: I am the international study chair and lead accruer for EA2192 as well as the study champion and lead national accruer for SWOG-2001. On the larger scale, I hold leadership roles within the Abramson Cancer Center (ACC) at the University of Pennsylvania and at the NCTN. I serve as the co-chair of the ECOG- ACRIN GI Committee, I am the co-leader of the ACC Cancer Therapeutics Program and I am the co-leader of the ACC Pancreatic Clinical Trial Program. These leadership positions are optimal platforms upon which to spearhead programs that (1) formally and longitudinally assist junior and mid-career oncology faculty in their quests to develop investigator-initiated studies and (2) develop a program at the NCTN that focuses on developing realistic trials for rare populations and then on accruing them successfully. To date, I have demonstrated a persistent and strong commitment to the NCI research enterprise. I have been involved with NCI-related research since my early career, initially attending meetings and later developing my own protocol, EA2192. Based on my steady engagement and input, I was appointed as the co-chair of the GI Committee in 2022 alongside Jordan Berlin (see LOS). Together, we have made a commitment to improving the process of clinical trial development, a mission that will be critical in order for the NCTN to remain competitive in an ever-changing landscape. Over the past three years, we have employed multiple initiatives including boosting the education of our investigators about the NCTN process, leaning on the excellent Working groups chairs to fine-tune concepts prior to Committee Presentation and providing substantial assistance during the submission process. With the support of the R50 Research Specialist award, I will build further on this approach in two ways: At the ACC, I will develop a sustainable program for early and mid-career investigators to provide longitudinal support in the development of investigator-initiated trials, with a particular focus on rare disease studies that can be best executed via the ECOG-ACRIN system. Within the NCTN, I will employ a novel program assisting investigators in the development and successful enrollment of trials for rare disease populations, a subset of studies that have lost ground since the COVID pandemic. Specifically, I will create a living resource for NCTN investigators that focuses on methods to design practical, feasible studies and will provide longitudinal support to those across the NCTN who are developing studies in this space. My ultimate goal is to develop and implement pragmatic clinical trials that address key questions in the field of gastrointestinal malignancies, particularly for patients with rare subsets of disease.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Sickle cell disease (SCD) and thalassemia are most common genetic disorders worldwide and cause early mortality and suffering. Reversal of the developmental hemoglobin switch to increase levels of fetal hemoglobin, encoded by HBG, reduces SCD complications. Previous work over several decades has elucidated several key transcription factors required to repress HBG expression, but they have proven difficult to target therapeutically with small molecules. Therefore, a deeper understanding of how cell type and context-specific gene expression programs are coordinated throughout development is essential to design therapies that reactivate HBG selectively while sparing other blood cell lineages. While transcription factor recruitment is considered a key control point for gene specificity, less is known about the selectivity of co-regulatory complexes. This proposal focuses on one such transcriptional co-activator complex, SAGA, and aims to disentangle its functions throughout human hematopoiesis. My overarching hypothesis is that despite its ubiquitous expression, SAGA has gene- and cell type-specific functions across hematopoiesis. Aim 1 elucidates its gene-specific roles using precision base editing approaches and functional genomics. I will fine-map SAGA subunit functions both genome-wide and on globin gene regulation during erythroid development and differentiation. Aim 2 investigates the distinct roles of SAGA amongst human blood cell lineages using both in vitro and in vivo xenograft model systems. The proposed research builds on my expertise in developmental hematology, genomics, and genome editing and provides a generalizable framework to dissect the functions of multi-subunit complexes and gene pathways. Through this detailed molecular mapping of the SAGA complex, I will examine how SAGA impacts specific loci in primary hematopoietic cells across both lineage and developmental state. While targeting ubiquitous factors has been viewed as therapeutically undesirable, this proposal revisits this concept by focusing on SAGA to discover new context-specific functions that could direct future precision therapies for SCD and other blood disorders.

NIH Research Projects · FY 2025 · 2025-08

Abstract Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related deaths and a significant global health concern. Early detection is crucial, as it significantly improves survival rates; patients diagnosed early can expect a median survival of over 60 months, compared to less than 15 months for those diagnosed at advanced stages. Unfortunately, most HCC cases are diagnosed late, underscoring the urgent need for developing better detection methods. The current sensitivity of ultrasound for early-stage HCC detection is only about 45%, which increases to 63% when alpha-fetoprotein (AFP) testing is added, indicating a pressing need for new imaging techniques to enhance detection capabilities. This research proposes a novel imaging method, hydralazine-augmented contrast ultrasound (HyCUS), which aims to improve HCC visibility by increasing the image contrast between tumors and surrounding tissue by differentially modulating their blood flow. The study will validate HyCUS using an autochthonous rat model of HCC with liver fibrosis, assessing visibility through objective computational metrics and subjective assessments by radiologists. The proposal has three specific aims. Specific Aim 1 will establish HyCUS as a technique for enhancing HCC visibility. The underlying principle of the technique will be validated by measuring the differences in blood flow between HCC and surrounding liver tissue after hydralazine administration. These differences will be correlated to Evans blue dye uptake to assess visibility objectively. Specific Aim 2 will develop an imaging regimen for HyCUS. Understanding the dose-response relationship of hydralazine is essential for clinical applications. These studies will investigate the effect of different hydralazine doses on the visibility of lesions, the timing of contrast enhancement, and the duration of this enhancement across various tumor sizes. Specific Aim 3 will evaluate the performance of HyCUS against existing methods (B-mode ultrasound and contrast-enhanced ultrasound) in detecting early-stage HCC and measuring tumor burden. These studies will track tumor formation and growth of HCC in a rat model, assessing the earliest detection times and overall tumor burden through necropsy analysis post-imaging. In conclusion, the proposed HyCUS technique represents a promising advancement in the early detection of HCC. Enhancing the visibility of tumors through targeted blood flow modulation could significantly improve diagnostic capabilities and ultimately improve patient outcomes in liver cancer management.

NIH Research Projects · FY 2025 · 2025-08

Summary: Ulcerative colitis (UC) is a subtype of inflammatory bowel disease with symptoms of abdominal pain and bloody diarrhea secondary to colonic inflammation. Human colonic mast cells (MCs) express a novel G protein-coupled receptor known as Mas-related GPCR-X2 (MRGPRX2, mouse ortholog, MrgprB2), and RNAseq analysis of inflamed UC samples demonstrated enhanced expression of MRGPRX2 agonists and MC proteases when compared to non-inflamed samples. It is well documented that GPCRs undergo desensitization following their phosphorylation at Ser/Thr residues. Interestingly, analyses of human GPCR protein-altering single nucleotide polymorphism (SNP) from exome chip data of inflammatory bowel disease showed that N62S (rs10833049) mutation of MRGPRX2, which results in the creation of a phosphorylation site, is associated with protective phenotype in UC. By contrast, we found that another SNP, in which a Ser is replaced with Leu (S325L) results in reduced desensitization and greater degranulation than the WT receptor. Based on these findings, we hypothesize that MRGPRX2 expressed in colonic MCs contributes to UC and that its phosphorylation and desensitization modulates disease outcome. Given that MrgprB2 is the mouse counterpart of human MRGPRX2, it is expected that MrgprB2−/− mice would display a reduced disease phenotype in experimental models of UC when compared to WT mice. Surprisingly, however, recent studies showed that MrgprB2 displays a protective effect in experimental UC in mice. This difference could reflect the fact that MRGPRX2 and MrgprB2 display low sequence homology (~53%) and suggests that mice expressing MrgprB2 may not adequately reflect the situation in humans. To overcome this limitation, we utilized CRISPR/Cas9-mediated gene editing approach to replace MrgprB2 with human MRGPRX2 (MRGPRX2-KI mice). We found that primary MCs from MRGPRX2-KI mice respond to agonists that are implicated in UC for substantially greater degranulation at lower concentrations than primary MCs from WT mice expressing MrgprB2. In aim 1, we will utilize MRGPRX2-KI, WT and MrgprB2-/- mice to test the hypothesis that, unlike MrgprB2, MRGPRX2 expressed in colonic MCs contributes to the pathogenesis of UC. In aim 2, retrovirus will be used to express MRGPRX2 and its phosphorylation variants N62S and S325L in MrgprB2−/− mouse bone- derived mast cells, which will then be engrafted into MC-deficient mice. This strategy will be used to determine how SNPs on MRGPRX2 modulate experimental UC. Successful completion of this study will lead to the characterization of a new preclinical model of UC and may provide novel insights on how individual variations in MRGPRX2 alters disease phenotype.

NIH Research Projects · FY 2025 · 2025-08

Title: Deciphering novel cell death mechanisms and metabolic reprogramming in response to microenvironmental stress. Understanding how cells respond to microenvironmental stress is essential for unraveling fundamental biological processes. While acute stressor perturbations in tissues are often transient, the cellular changes induced can persist long after the initial stressor has subsided, challenging our current knowledge of long- term impact of microenvironmental stress. My laboratory combines cell- and genome-engineering with transcriptional, epigenetic, and metabolic profiling to understand cellular fate decisions informed by the surrounding microenvironment. With these tools, we have revealed an unexpected phenomenon: Cells exposed to immune cell activation can undergo cell death without specific antigen recognition, cytokine signaling or apoptosis induction, suggesting the existence of a novel mechanism of cell death with far-reaching implications for various biological processes. Our preliminary experiments provide a crucial insight: the disruption of translation of mitochondrial respiratory chain genes confers resistance to this form of cell death in exposed cells. This observation suggests that exposure to immune activation induces nutrient deprivation in the surrounding microenvironment, forcing exposed cells to adopt alternative metabolic pathways for survival. The absence of widespread tissue death during exposure to immune activation further supports this hypothesis, indicating that surviving cells likely undergo significant metabolic reprogramming to adapt to these challenging conditions. The research in this Maximizing Investigators' Research Award proposal aims to capitalize on our deep expertise in cell engineering and metabolic profiling to (I) characterize the precise mechanism of cell death in non-targeted cells exposed to immune activation and (II) elucidate the metabolic and subsequent epigenetic reprogramming events occurring in surviving cells, along with their long-term consequences in the contexts of wound healing, cellular senescence, and tissue homeostasis. Understanding this novel cell death mechanism and its evasion through metabolic reprogramming has the potential to revolutionize our comprehension of fundamental cellular processes. This knowledge could have profound implications for diverse fields of cell biology, shifting paradigms in our understanding of how cells respond to external stressors and commit to life-or-death fate decisions. Our findings may illuminate new principles of metabolic plasticity and cellular adaptation, providing insights into the complex interplay between metabolism, cell survival, and cell death pathways. Ultimately, this research will expand our basic understanding of cellular homeostasis, stress responses, and tissue maintenance, contributing to the broader knowledge base of cellular and molecular biology. These fundamental insights could inform future studies across various biological disciplines, potentially opening up new avenues for exploring cellular behavior in both normal and pathological states.

NIH Research Projects · FY 2025 · 2025-08

Engineered immunotherapies harness the power of the immune system to combat diseases and to attack specified targets, such as cancer cells. While their applications have been transformative, a major challenge toward widespread use is the presence of sustained stimuli in a tissue or tumor microenvironment that strongly inhibit immune cell function. Chronic inflammatory signaling via interferons causes phenomena such as T cell exhaustion and macrophage phenotypic switching. New platforms that enable cells to robustly maintain their effector functions, despite sustained inflammatory hyperstimulation from interferons, are urgently required to drive the field forward. To address this critical unmet need, we propose development of synthetic compartments capable of insulating and sequestering signaling components. This strategy can accelerate reaction rates and rewire signaling, altering flux through the Type I and Type II interferon pathways to restore immune cell competence. We have developed a modular intracellular microcompartment platform based on the self-assembly of specific intrinsically disordered proteins (IDPs). Using this system, along with tagged enzymes, we have demonstrated selective and robust control of component sequestration and release, enabling on demand control of cell behaviors. In preliminary data, we have demonstrated the feasibility of targeting janus kinases (JAKs) to synthetic compartments, feasibility of efficiently sequestering TYK2, feasibility of expressing condensates in human T cells, and feasibility of generating knockouts of TYK2 with reduced Type I signaling. In Aim 1, we propose to further develop this platform to ‘insulate’ and shut down key components of JAK STAT signaling in Type I interferon pathway, which is known to result in T cell exhaustion. We will target and functionally insulate the kinase TYK2 via inducible expression of our synthetic compartments. We will also measure immune-related pathways as validation that our approach is feasible without major alteration of T cell physiology. In Aim 2, we propose to redirect repressive Type I signals to Type II cell stimulatory outputs, effectively reprogramming intracellular inflammatory signaling. In Aim 3, we will use these strategies in an in vitro co-culture model of mouse melanoma and T cells to restore immune cell function under conditions that cause exhaustion. We will test the extent to which transient sequestration of TYK2 shuts down interferon Type I signaling, enabling T cell cells to block tumor cell growth. The future of personalized medicine depends on engineering syngeneic cells that target disease tissues and are resistant to deactivating signals. By harnessing the power of compartmentalization and controlled release, our platform provides a new and versatile toolkit to program immune cell responsiveness for next-generation cell-regenerative therapies.

NIH Research Projects · FY 2025 · 2025-08

The overall goal of this application is to elucidate the role of TRIM10 in maintaining solubility and function of TDP-43 and protecting against TDP-43 proteinopathies. TDP-43 is an RNA-binding protein that resides predominantly in the nucleus of healthy cells, regulating various aspects of RNA metabolism. In progressive and fatal neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), limbic predominant age-related TDP-43 encephalopathy (LATE), and other Alzheimer's disease-related dementia (ADRD), TDP-43 often misfolds and aggregates in the cytoplasm of affected neurons and glia, which is accompanied by its depletion in the nucleus. Moreover, mutations in TDP-43, which increase its propensity to misfold and aggregate, are associated with a familial subset of ALS and FTD. In both sporadic and familial TDP- 43 proteinopathies, aggregation of TDP-43 largely occurs in an age-dependent manner, suggesting a diminished capacity of a cellular factor(s) that can suppress TDP-43 aggregation early in life. However, the identity and nature of such a factor(s) remains unknown. This conspicuous gap in our knowledge impedes the development of effective therapies. To maintain their proteins in the functional soluble form, organisms in all kingdoms of life rely on protein quality control (PQC) systems. Over the past decade, research in my lab has suggested that animal cells may have a unique and highly potent PQC system consisting of tripartite motif (TRIM) proteins. We found that individual TRIM proteins possess multiple activities to enable protein folding, and they operate via a mechanism distinct from canonical ATP-dependent PQC systems. More recently, we demonstrated that TRIM11 is highly effective in preventing tau aggregation. Our preliminary data further showed that another TRIM (TRIM10) may potently suppress TDP-43 misfolding and aggregation, and its expression may be markedly downregulated in TDP-43 proteinopathies. Here, we propose to test the central hypothesis that TRIM10 is critical for maintaining TDP-43 in its functional soluble state, and downregulation of TRIM10 contributes to the pathogenesis of TDP- 43 proteinopathies. We will (1) characterize the degradative, chaperone, and disaggregase activities of TRIM10 towards TDP-43, (2) determine the effect of TRIM10 on phase behavior and function of TDP-43, and (3) define the role of TRIM10 in the pathogenesis of TDP-43 proteinopathies and explore its utility in disease treatment. Collectively, these studies will enrich our understanding of protein homeostasis and TDP-43 proteinopathies and inform the development of effective therapies.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Artificial intelligence (AI) is poised to transform healthcare, particularly in clinical decision support for treatment selection in a range of diseases. However, AI is underutilized in clinical care, in part due to two unique challenges: 1) noisy and complex health data, such as multi-modal electronic health record (EHR) data, and 2) demand for a high degree of trustworthiness in treatment selection use cases. Explainability is a core principle of trustworthy AI, but may be lacking in current state of the art (SOTA) AI models for healthcare, particularly for analysis of multi-modal EHR data. Most notably, current approaches to explainable AI in healthcare based on post-hoc explanations of black-box models are unlikely to offer clinically meaningful and actionable explanations to clinicians and end-users, thereby decreasing trust and buy-in. Furthermore, treatment effect estimation and selection based on inaccurate explanations may cause patient harm. As such, there is an urgent need for inherently explainable AI/ML models in healthcare that also enjoy the robustness and expressivity of modern deep learning models. Our preliminary work suggests that incorporation of explanations based on existing clinician domain knowledge would be a key facilitator for trusting AI models that predict treatment benefit. However, to the best of our knowledge, no existing SOTA AI models effectively incorporate clinical domain knowledge into explanations. To address these gaps, we propose to develop Med-Scallop, a novel methodology and associated software tool based upon the emerging paradigm of neurosymbolic AI. This paradigm can effectively integrate deep neural models with symbolic domain knowledge from human experts (e.g. physicians), yielding neurosymbolic programs that are more robust, data-efficient, and interpretable than their neural or symbolic counterparts alone. Our preliminary quantitative and qualitative results suggest that Med-Scallop can generate clinically meaningful explanations for clinical decision support (CDS) and facilitate AI-physician collaboration. We propose to assess the performance of our proposed neurosymbolic AI models using two clinical use cases for guiding optimal individualized treatment selection and management for patients with non-small cell lung cancer and sepsis, using multi-institutional EHR datasets for training and validation. Our specific aims are to 1) develop inherently explainable neurosymbolic AI architectures for treatment selection using temporal EHR data; 2) integrate multi-modal clinical data into a neurosymbolic framework for faithful and accurate treatment selection; and 3) develop Med-Scallop, an open-access software platform for neurosymbolic AI for CDS, as well as a comprehensive mixed-methods methodology for assessing clinical validity and usability of explanations from AI models. Our approach represents a new, transformative approach to harness the benefits of both deep neural networks and interpretable symbolic models without suffering their limitations and the power of rich yet complex EHR data.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT: Neurodevelopmental disorders (NDDs) represent a diverse array of conditions characterized by abnormal brain development and function which cause motor, cognitive, and communicative deficits that greatly impact a patient’s quality of life. However, many underlying disease mechanisms are still unknown, resulting in a dearth of effective treatments and an urgent need to better understand NDDs. One challenge of studying NDDs and developing effective treatments however is that many NDDs are multifactorial with both an individual’s genetics and environment playing key roles. Attributing a single disease mechanism to a patient phenotype can therefore be extremely difficult. Consequently, one approach to circumvent this is by focusing on NDDs with a singular and well-defined genetic cause. To this end, the Akizu lab recently identified a novel NDD caused by heterozygous missense mutations in Enhancer of Zeste Homologue 1 (EZH1) which is one of the two catalytic subunits within the Polycomb Repressive Complex 2 (PRC2) that is responsible for transcriptional repression through tri-methylation of the lysine at position 27 of histone H3 (H3K27me3). Initial mechanistic work identified several mutations that increase the catalytic activity of EZH1 and subsequent H3K27me3 levels, indicative of a gain-of-function (GOF) effect. Follow-up experiments further revealed that one GOF mutation p.A678G induces premature neuronal differentiation and enhanced maturation during the differentiation process along with a downregulation of glutamatergic synapse genes that together may potentially produce dysfunctional neurons. However, several unanswered questions remain such as whether the remaining 10 uncharacterized EZH1 missense mutations are GOF, if the previously characterized phenotype extends to other validated GOF mutations and also induces neuronal dysfunction, and whether EZH1-specific inhibitors can ameliorate these effects. This proposal therefore aims to answer these questions and gain further insight into the EZH1 GOF phenotype and relevant treatment strategies, advancing our understanding of NDD disease mechanisms and therapeutic approaches to help afflicted patients. The proposed research will also provide the applicant with intensive training in imaging and molecular methodologies; rigor and reproducibility; experimental design; and scientific communication. Furthermore, the applicant will be supported by a team of experts headed by the sponsor Dr. Naiara Akizu and also including Dr. Ethan Goldberg, Dr. Ana Cristancho, and Dr. George Burslem. Finally, the resources available at both the University of Pennsylvania and the Children’s Hospital of Philadelphia will ensure the successful completion of the proposed research and training of the applicant and will help fulfill their desire to become an independent research professor of neuroscience.

NIH Research Projects · FY 2025 · 2025-08

Proposal Summary/Abstract Candidate: Dr. Leo Wang holds a BA, MS, MD, and PhD from the University of Pennsylvania, where he completed dermatology residency and currently is a Clinical Instructor and postdoctoral fellow working with Dr. George Cotsarelis in the Department of Dermatology. In 2025, Dr. Wang will be appointed as Assistant Professor in the tenure track and provided a generous startup and laboratory space as an independent researcher. Support from the DP5 award will enable the pursuit of the proposed research in Dr. Wang’s future laboratory. Environment: The proposed research will be conducted in the Department of Dermatology at the University of Pennsylvania, which is committed to Dr. Wang’s career development and mentorship. The Department of Dermatology is consistently top ranked in NIH funding with the support of a NIH P30 Skin Biology and Disease Resource-based Center, which enhances research and collaboration across the University. The School of Engineering and Applied Sciences is a block away from the School of Medicine, allowing Dr. Wang to maintain multidisciplinary collaborations. Research: Human skin wounds typically heal with fibrotic scars, leading to impaired form and function, characterized by the loss of appendages like hair follicles. Scarless wound healing requires hair follicles, which are rich in stem cells and factors that can prevent and treat scars. While humans do not regenerate hair follicles, mice do so in large full-thickness wounds through wound-induced hair neogenesis (WIHN), involving sequential upregulation of Wnt and Sonic hedgehog (Shh) signaling. This proposal aims to spatiotemporally activate Wnt and Shh signaling using a two-component hydrogel that releases CHIR99021, a Wnt agonist, and HhAg1.5, a Shh agonist, to promote hair follicle neogenesis and treat scars. Hydrogels will be engineered with microcapsules for tunable release of CHIR and HhAg, tested to induce WIHN and prevent scarring, and fabricated into microneedle patches to simultaneously wound and induce hair neogenesis, and treat fibrotic, mature human scars. This research will demonstrate that biomaterials can control exogenous signals to prevent and treat scars through hair follicle neogenesis, advancing the understanding of WIHN and expanding approaches to controlled delivery. Most importantly, this will lead to a therapeutic that can be injected into skin after surgery or trauma to prevent scars, or applied as patches to treat mature scars, which could transform clinical care.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Studies of public health interventions aimed at controlling the spread of infectious diseases such as HIV/AIDS or COVID19 often face important methodological challenges due to pervasive network dependence, confounding and widespread missing data. Each of these complications has separately received considerable attention, however, methods to tackle them when they coexist are currently lacking. We will develop new statistical methodology, specifically causal identification theory and robust estimation theory which we plan to apply to address pressing scientific questions in infectious disease research using data from two randomized trials and two observational studies with data at hand, where both missing data, and network structure occur including the HAALSA South African Study, the Networks, Norms, and HIV/STI Risk Among Youth (NNAHRAY) study, and the Botswana Combination Prevention Study (BCPP) and the Home-based Interventionto Test and Start (HITS) cluster randomized trial. Success in the proposed research will not only allow for robust conclusions to be drawn from data in the above studies, despite the presence of missing data, the potential for confounding bias, and complex social network structure; it will also provide a methodological template for addressing similar questions beyond these four studies as confounded, missing and dependent data are routinely co-occurring complications in Social and Infectious Disease Epidemiology.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY βII-spectrin, a key component of the cytoskeleton, forms a submembrane lattice that organizes micron-scale domains of transmembrane proteins. βII-spectrin also facilitates organelle transport and likely provides structural support and flexibility to neurons. Loss of neuronal βII-spectrin in mice causes early postnatal lethality and significant alterations in cortical development and axonal connectivity. Heterozygous variants in SPTBN1, the gene encoding βII-spectrin, cause an early-onset neurodevelopmental syndrome characterized by speech and motor delays, comorbid with intellectual disability, autism spectrum disorder, ADHD, epilepsy, dysmorphisms, and cortical deficits, collectively known as SPTBN1 syndrome. Previous structure-function evaluations of disease variants in heterologous cells and rescue studies in βII-spectrin null murine cortical neurons indicate that pathogenic mechanisms involve βII-spectrin instability and aberrant binding to key molecular partners. My preliminary data shows that missense and nonsense SPTBN1 syndrome variants differentially impact βII- spectrin protein levels and subcellular localization. These findings imply that different groups of βII-spectrin variants impart neuronal dysfunction through divergent mechanisms. However, the molecular pathways through which these variants affect the total βII-spectrin pool and downstream βII-spectrin-dependent processes in human neurons remain unexplored. In this study, I will evaluate the premise that nonsense and missense SPTBN1 syndrome variants disrupt βII- spectrin function in human cortical neurons (hCNs) through molecularly distinct pathological changes in expression and subcellular distribution. I will first study nonsense SPTBN1 variants in hCNs to determine the molecular and functional consequences of pathogenic βII-spectrin haploinsufficiency compared to βII-spectrin heterozygous knockout hCNs (Aim 1). I will also interrogate missense SPTBN1 syndrome variants that distinctly change βII-spectrin levels and distribution, uncover mechanistic basis for these effects, and explore methods to ameliorate βII-spectrin misexpression (Aim 2). These experiments will leverage the power of endogenous expression of disease-linked SPTBN1 variants at heterozygous levels in hCNs to uncover mechanistic insights into SPTBN1 syndrome pathogenesis. Ultimately, this work will guide the future design of therapeutics for SPTBN1 syndrome and other spectrinopathies of the nervous system.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Wilms tumor is the most common pediatric renal tumor, primarily affecting children under the age of 5. It is closely linked to disrupted nephrogenesis and features rudimentary structures found in the developing kidney. While alterations in transcription factors and signaling proteins crucial for kidney development are known molecular drivers, recent studies have revealed that 30-50% of Wilms tumor patients have mutations in epigenetic regulators, with ENL (Eleven-Nineteen Leukemia) being the most frequently mutated. The role of these epigenetic regulators in kidney differentiation and tumorigenesis remains unexplored. This study aims to fill this gap by exploring the role of ENL mutations in nephrogenesis and Wilms tumor development. ENL is a chromatin reader protein which binds histone acetylation through its YEATS domain to recruit co-factors and drive transcriptional elongation. Hotspot mutations in the ENL YEATS domain have been identified in Wilms tumor. These mutations result in hyperactivation of target gene transcription by the formation of high local concentration assemblies termed condensates. To study the role of ENL mutations in Wilms tumor, our lab developed a conditional knock-in mouse model for the most frequent Enl mutation, referred to as Enl-T1. We crossed these mice to the Wt1-Cre strain to induce heterozygous expression of Enl-T1 in the nephron and stroma lineages of the developing kidney. These mice displayed aberrant kidney development with the loss of differentiated nephron structures and post-natal lethality. Single-cell analyses of transcriptomics and open chromatin accessibility revealed that Enl-T1 shifts nephron progenitor cells toward an aberrantly committed state while restricting their further differentiation. Enl-T1 expression also disrupted stromal progenitor cell differentiation, suggesting a role for ENL in both stromal and nephron progenitor populations. Notably, the developmental defects and transcriptional changes induced by Enl-T1 can be rescued by treatment with TDI-11055, an inhibitor that blocks the interaction of ENL-T1 with acetylated histones. Based on these findings, I hypothesize that an optimal level of ENL activity is critical for normal kidney development, and disruptions to this balance through gain-of-function mutations in ENL can lead to tumor formation. I will investigate the role of the Enl-T1 in nephrogenesis and tumorigenesis using lineage specific cre drivers (Aim 1) and determine the impact of the ENL mutant inhibition, using TDI-11055, on Wilms tumor maintenance (Aim 2) through a combination of mouse modeling, histological analysis, single cell transcriptomics, and patient-derived Wilms tumor organoids. These studies will elucidate the role of Enl mutations in kidney development and disease, identify a novel therapeutic target in Wilms tumor, and provide new insights into how epigenetic regulation plays a role in cell differentiation in normal and diseased contexts.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Bronchopulmonary dysplasia (BPD) is the most common complication affecting preterm neonates and is defined by supplemental oxygen dependence at 36 weeks postmenstrual age. Advances in neonatal care have improved overall survival of preterm neonates though BPD prevalence continues to increase. BPD develops after early-life lung injury caused by myriad pre- and postnatal factors such as the administration of supplemental oxygen. Though a lifesaving therapeutic intervention, high concentrations of oxygen (hyperoxia) arrest alveolarization and vascularization and induce inflammation. Neonates with BPD face challenges in lung function that can persist throughout adulthood. BPD has a striking male bias for both incidence and severity, demonstrating the need to interrogate sex as a biological variable in lung development, injury, and repair processes. Previous work from our lab showed that female chromosome complement (XX) is protective against lung injury, independent of gonadal sex (ovaries and testes). It is unknown if the protective effect is due to the presence of two X chromosomes or the absence of a Y chromosome. The goal of this proposal is to interrogate dosage sensitivity of sex chromosomes and an X-linked gene, Kdm6a, in an early-life hyperoxia mouse model of BPD. I hypothesize X chromosome mechanisms and dosage contribute to female resilience against hyperoxic lung injury. This hypothesis will be tested by the following: Aim 1) Determine the role of sex chromosome dosage in neonatal hyperoxic lung injury and Aim 2) Determine the role of endothelial Kdm6a in neonatal hyperoxic lung injury. In Aim 1, I will assess lung morphometry and vascularization in room air and hyperoxia-exposed in the XY star (XY*) mouse model that has XX, XO, XY, and XXY genotypes. My preliminary data shows modulation of alveolarization after injury as a function of chromosome dosage. I will then determine cell-type specific gene signatures after hyperoxia exposure using single-cell RNA sequencing. In Aim 2, I will determine the role of endothelial Kdm6a in female neonatal mouse pups during development and after hyperoxic lung injury. X chromosome inactivation (XCI) is a dosage compensation mechanism whereby supernumerary X chromosomes are transcriptionally silenced so genes from only one X chromosome are expressed. However, some genes escape XCI. Intriguingly, preliminary data from our lab show a female- specific induction of an XCI escapee, Kdm6a, after exposure to hyperoxia. I will test the hypothesis that endothelial knockdown of Kdm6a intensifies hyperoxic lung injury in female mice. To determine the role of Kdm6a in epigenetic regulation and alterations to the chromatin landscape, I will perform bulk RNA-seq, ATAC- seq, and CUT&RUN. Completion of this proposal will advance mechanistic knowledge of sex-specific responses to respiratory insults and potentially inform the development of precision medicine approaches.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Digital health interventions (DHIs) have shown significant promise in promoting HIV prevention and care among at-risk youth. Despite the great potential for behavior change that DHIs offer, the effectiveness of DHIs has varied across studies, largely due to the varying degrees of participant engagement. Therefore, identifying key engagement characteristics in DHIs is important to evaluate the efficacy of DHIs with rigor and enhance the efficacy of DHIs for at-risk youth by promoting engagement. DHI studies have utilized single indicators of engagement, such as amount, frequency, and duration, and have focused on cross- sectional relationships between engagement and outcomes. This approach has limited the ability to evaluate the multidimensional aspects of engagement and the prospective effects of engagement on outcomes over time. Moreover, the one-size-fits-all approach may not be suitable for designing DHIs for youth. Therefore, there is a need to analyze engagement across multicenter research trials dedicated to HIV prevention and care for youth in the US. Our research aims to identify key characteristics of engagement to enhance the efficacy of DHIs for at-risk youth by pooling data from Adolescent Medicine Trials Network for HIV Interventions (ATN) - UNC/Emory Center for Innovative Technology (iTech) trials. In this study, we propose using secondary data from more than 900 individuals who participated across five ATN-iTech trials (MyChoices, LYNX, TechStep, Get Connected, and P3). The study's specific aims are: 1) To harmonize archived paradata into common data elements of participants' DHI engagement across ATN trials; 2) To identify engagement typologies and examine their associations with HPC outcomes; and 3) To examine longitudinal associations between participants’ prospective engagement and desired HPC outcomes over time. This research significantly contributes to current scientific knowledge by detangle the complex relationship between engagement and DHI outcomes to optimize DHIs for youth by pooling a multicenter research network in the U.S. Our findings will help to understand the characteristics driving higher engagement, rigorously evaluate the efficacy of DHIs while considering engagement, and propose optimized strategies to promote engagement, ultimately enhancing the efficacy of DHIs for youth. These insights will provide a blueprint for researchers to rigorously evaluate DHIs and enhance their effect sizes for adolescents and young adults at risk for HIV and inform the development of an implementation science framework for engagement in digital HIV interventions, which may be used in future studies for at-risk youth.

NSF Awards · FY 2025 · 2025-08

Mechanical metamaterial, i.e., engineering materials whose properties arise from geometry rather than chemical composition, hold great promise for applications in aerospace, biomedical engineering, robotics, and beyond. By integrating active elements, these materials can respond dynamically to environmental stimuli such as heat or light, enabling adaptive and programmable behavior. Despite their potential, the practical design of such materials remains constrained by computationally expensive and highly specialized modeling tools. This award supports fundamental research to establish a general, experimentally validated continuum modeling framework for active mechanical metamaterials. This framework will enable efficient prediction of material behavior and systematic design across a wide range of geometries and actuation mechanisms. The project advances fundamental understanding while supporting technological innovation in advanced manufacturing, adaptive devices, and reconfigurable structures. It also leverages interdisciplinary collaboration and integrated fabrication-modeling workflows to train students across educational levels and to engage the public through outreach initiatives. The research integrates theoretical, experimental, and computational approaches. A continuum modeling framework will be constructed to connect unit-cell geometry with macroscopic deformation, including both planar and three-dimensional behaviors. The models will incorporate internal variables and compatibility conditions to capture soft deformation modes and the effects of actuation. Experimental work will validate the framework through fabrication, including direct ink writing, molding, and conventional 3D printing, and mechanical testing of passive and active metamaterials, making use of digital image correlation to quantify deformation. A systematic scaling study will define the limits of continuum applicability. The final phase will implement the models into finite element simulations to enable inverse design, allowing metamaterials to be tailored for specific responses. Collectively, these efforts will yield general design principles and computational tools to accelerate the adoption of mechanical metamaterials in advanced engineering applications. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.