Utah State Higher Education System--University Of Utah

NIH Research Projects · FY 2025 · 2025-09

Project Summary: Cardiovascular disease has consistently been the leading cause of death in the US for the last century with resultant heart failure (HF) accounting for nearly 800,000 deaths per year. Despite substantial advancements in clinically available interventions, the incidence of HF continues to rise with a staggering estimated economic burden of >$900 billion per year by 2030. Virtually all etiologies of cardiovascular disease leading to both heart failure with reduced ejection fraction (HFrEF) and preserved ejection fraction (HFpEF) involve pathological remodeling of the left ventricle characterized by excessive deposition of extracellular matrix (ECM) proteins that impair ventricular compliance and perpetuate HF pathogenesis. The primary cell type responsible for synthesizing and secreting ECM proteins are resident cardiac fibroblasts (CFBs) that become activated by diverse stimuli during HF. As a testament to the secretory potential of CFBs, a single fibroblast can produce >500,000 procollagen chains per hour which necessitates the presence of a robust network of protein folding machinery in the endoplasmic reticulum to maintain cellular proteostasis and allow for the proper processing of nascent ECM proteins. Such protein folding demands trigger activation of the unfolded protein response (UPR), an adaptive component of the proteostasis network to facilitate proper protein quality control. Our long-term goal is to determine the mechanistic contributions of the UPR in regulating chronic CFB activation and reactive cardiac fibrosis in response to HFrEF and HFpEF. Our preliminary data support the notion that the IRE1 arm of the UPR plays a pivotal role in protecting against fibrotic remodeling in the heart via targeted mRNA degradation of transcripts encoding proteins required for CFB activation, namely Tmem100. We’ve also characterized a novel small molecule activator of IRE1 with potential to ameliorate functional decline in a preclinical model of HFrEF. In this proposal, we will focus on IRE1 and whether tactile control of the endonuclease activity of IRE1 could alter CFB activation and ECM deposition with the hypothesis that IRE1 protects against pathological cardiac fibrosis in HF via regulating the selective degradation of Tmem100 and a pro-fibrotic transcriptome. We will address this hypothesis using fibroblast-specific gene targeting in complimentary mouse models of HF, as well as mechanistic studies in primary CFBs, in the following Specific Aims which are to: (Aim 1) determine the cardiac fibroblast transcriptomic profile regulated by IRE1 using a mouse model of HFrEF, (Aim 2) determine the functional significance of IRE1-mediated degradation of Tmem100 mRNA in fibrotic remodeling using a mouse model of HFrEF, and (Aim 3) evaluate the therapeutic efficacy of novel small molecule activator of IRE1 in mitigating fibrotic remodeling using mouse models of HFrEF or HFpEF. These studies are significant as they present the opportunity to identify novel mechanisms contributing to CFB activation and ECM deposition as well as to test new therapeutic strategies with potential to ameliorate fibrotic remodeling associated with both HFrEF and HFpEF.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY The transformative discovery of immune checkpoint inhibitors (ICIs) has unlocked antitumor immune responses in a wide range of cancer patients. Nonetheless, alternative treatment options are critically needed for the majority of patients who are non-responders to ICIs. Even cancer types known to have the highest response rates such as Hodgkin lymphoma can cause relapsed or refractory disease in a subset of patients. Among the key factors hindering the clinical benefit of ICIs are T cell exhaustion and suppressive myeloid cells. Exhausted T cells can be locked in a dysfunctional state by multiple redundant mechanisms including the suppressive action of myeloid cells. Despite the documented clinical relevance of suppressive myeloid cells, however, targeted therapies remain in clinical development. The Research Plan addresses the hypothesis that JAK inhibitors in conjunction with checkpoint inhibitors convert myeloid cells from a suppressive into an immunostimulatory state, inducing clinical responses in ICI monotherapy-resistant patients. The proposed studies leverage concordant preliminary preclinical and clinical preliminary data from preclinical cancer models and a clinical trial in Hodgkin lymphoma patients with high efficacy. In Aim 1, key mechanistic features of the effects of the JAK inhibitor ruxolitinib will be determined. Aim 2 focuses on the identification of cellular and molecular correlates of response to the combination therapy of ruxolitinib with the ICI nivolumab in Phase 1b and II clinical trials of relapsed or refractory Hodgkin lymphoma, including evaluation of the hypothesis that responders exhibit more baseline circulating suppressive myeloid cells than non-responders. In Aim 3, the cell- intrinsic role of JAK1 and JAK2 in this combination therapy will be established, and the hypothesis that MHC class I-independent responses are preferentially enhanced will be tested. The principal investigator Dr Zak is an immunologist with expertise in chemical biology, computational biology and clinical studies, possessing a comprehensive toolkit to execute this translational project. His long-term goal is to establish an independent research program centered on identifying and targeting immunosuppressive pathways in cancer. Dr Zak has established a clinical collaboration enabling the development of a novel combination therapy. A comprehensive career development plan addresses four key areas of training to enhance the launch of his research program: (1) utilizing relevant models of cancer, (2) design, management and analysis of collaborative clinical studies, (3) grantsmanship and progression towards independent grant submission and (4) establishing a laboratory, hiring, mentoring and management. The career development activities will forge the skills needed to launch and sustain an independent academic research program in cancer immunology.

NIH Research Projects · FY 2025 · 2025-09

Summary The mutual interactions between the host and the gut microbiota confer health benefits to the host and provide a nutrient-rich environment for the microbial community. However, our knowledge of the direct functional impact of host factors on the gut microbial community remains incomplete. The monolayer of intestinal epithelial cells (IECs) provides the front-line response to the luminal contents, including the gut microbiota, for maintaining intestinal homeostasis. Our previous works and preliminary data have demonstrated a previously unrecognized function of IECs on the gut microbiota: IEC-derived components promote Lactobacillus rhamnosus GG (LGG) growth, the protective effects of LGG on IECs, and production of p40, an anti-inflammatory factor, thereby ameliorating colitis in mice. To elucidate the mechanisms involved in this action, we have used novel transgenic mouse models to demonstrate that IEC-EVs interact with a wide range of microbiota, in addition to Lactobacilli, in the mouse gastrointestinal (GI) tract, and blocking extracellular vesicles (EV) release by IECs increases inflammatory disease severity consistent with the importance of IEC-EVs for enforcing gut homeostasis. We further find that IEC-EVs could be up-taken by LGG and promote LGG growth and protective effects on IECs. Thus, IEC-EVs may functionally mediate the communication between IECs and the gut microbiota. Importantly, we have identified a molecular chaperone, heat shock protein (HSP) 90, particularly HSP90β as a high abundant cargo in IEC-EVs. Blocking HSP90 activity inhibits the increase in LGG growth and p40 production by IEC- derived components and IEC-EVs, indicating that IEC-derived HSP90β has potential clients with functionality in LGG. HSP90 is highly conserved from bacteria to mammals and displays functional overlap. Thus, HSP90β in IEC-EVs is a potential cargo to regulate growth and function of bacteria in the GI tract. We will test the hypothesis that IEC-EVs deliver functional protein cargos, including HSP90, to IEC-EV- targeted bacterial strains in the GI tract to promote their growth and functions in preventing intestinal inflammation. Studies in Aim 1 are proposed to determine whether HSP90 serves as a functional cargo in IEC- secreted EVs to promote LGG growth, promote the beneficial effects of LGG on IECs, and reinforce the efficacy of LGG for ameliorating colitis. Clients of HSP90 and HSP90-regulated pathways in LGG for LGG growth and function will also be investigated. In Aim 2, we will use novel mouse models to identify the microbial profile that are targeted by IEC-EVs and their contents to determine how extensive this mode of host-microbial crosstalk is in mammals during health and its contribution to preventing colitis in mice. Studies in this proposal will advance the field of microbial host mutualistic interactions by defining a novel crosstalk between the microbiota and host via IEC-EVs. Such knowledge will lay a foundation for developing strategies to enhance the effects of health- promoting commensal bacteria such as probiotics for preventing and treating inflammatory bowel disease.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT Globally, millions of pregnancies per year end in stillbirth, which occurs in 5.48/1000 U.S births, exceeding most high-resource countries and presenting major opportunities for improvement. Critical gaps in stillbirth risk stratification hamper efforts to accurately target interventions. Moreover, significant inequities persist; for example, American Indian/Alaska Native (AIAN) and Native Hawaiian/Pacific Islander (NHPI) people have 1.5- 2 times higher rates of stillbirth compared with White people. Further, stillbirth causes and experiences in AIAN and NHPI communities remain poorly elucidated. The University of Utah (UofU) is an ideal site for the NICHD “Road to Prevention of Stillbirth” Consortium, intended to reduce stillbirth in the U.S. For over 40 years, the UofU has led one of the most productive U.S. stillbirth research programs, with one of the few clinical programs dedicated to stillbirth prevention, staffed by leading experts with a history of national and global collaboration with clinicians, scientists, parents, and community groups. We recently launched the UofU Stillbirth Center of Excellence (USCoE), the first such center in the nation. With partners such as the University of Hawai’i, Tséhootsoí Medical Center (Navajo Nation), and Australia’s Stillbirth Centre of Research Excellence, we are poised to make innovative progress in preventing stillbirth and mitigating inequities. We also have a long history of effective participation in research (e.g. NICHD Maternal-Fetal Medicine Units (MFMU) Network, Stillbirth Collaborative Research Network, and Centers for Disease Control and Prevention), making UofU an ideal research network center. In addition to being an exceptional center for the Consortium, we propose to assess risk factors for adverse outcomes associated with decreased fetal movement (DFM). DFM is a major risk factor for stillbirth, occurring in up to 50% of cases. However, it also occurs frequently in normal pregnancies ending in live birth. Thus, risk stratification tools have great potential to improve stillbirth risk detection. We also aim to assess and address disproportionate stillbirth burdens in AIAN and NHPI communities. We will use audits to identify granular causes of and contributors to stillbirth through rigorous cause of death audits and qualitative approaches. Further, we will qualitatively assess parental experiences with stillbirth, related medical care, bereavement support, and evaluation for stillbirth. Disparities in these communities are unacceptable and have not been previously studied in a rigorous fashion. The UofU’s ability to support and complete Consortium research, in addition to our development of projects with high impact on stillbirth prevention and support, will ultimately reduce stillbirths and improve health equity.

NIH Research Projects · FY 2025 · 2025-09

Project Summary / Abstract This supplemental application to the NIDDK R25 Native American Research Internship (NARI) program proposes support for an Academic Career Development Workshop at the 2025 Association of American Indian Physicians (AAIP) Annual Meeting. The workshop will provide NARI alumni and American Indian and Alaska Native (AI/AN) undergraduate and post-baccalaureate students with research education resources as they pursue medical and graduate school, and early-career development as physician scientists and biomedical researchers. The workshop is designed to offer targeted mentorship, culturally grounded career guidance, and exposure to NIH-supported research pathways. It will build on the success of the 2024 AAIP Academic Career Development Workshop, which demonstrated strong engagement and positive outcomes among AI/AN trainees. The 2025 program will feature sessions led by NIH leadership, Indigenous faculty, and NARI alumni. Topics will include mentorship strategies, graduate and medical school preparation, NIH funding mechanisms, and research careers in diabetes, obesity, and metabolism. Participants will engage in roundtable discussions, keynote presentations, and interactive networking opportunities designed to foster long-term engagement with biomedical research and strengthen connections across NARI alumni cohorts. The supplement will support travel, lodging and per diem for NARI alumni and other eligible AI/AN students, participation by NARI staff, and evaluation of workshop outcomes. Interactive discussions with AI/AN physician leaders and NIH program officers will promote culturally sensitive dialogue and peer mentorship. This initiative complements the parent R25 award by expanding its reach and impact, strengthening the pathway of AI/AN scientists, and advancing NIDDK’s commitment to increasing AI/AN representation in the biomedical workforce. It will also serve as a model for integrating research career education with Indigenous values and community engagement.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Maturation during the HIV life cycle consists of the viral Gag and Gag-Pol polyproteins undergoing sequential proteolysis to create mature proteins, which organize into a functional viral core having the viral genomic RNA (gRNA) condensed as a ribonucleoprotein particle (RNP) inside the mature capsid. Eccentric condensate (EC) phenotypes, consist of the RNP localized outside the capsid, lead to loss of infectivity. The EC phenotype is caused by antivirals that target both the viral CA and IN proteins; additionally, this has been shown in so-called class II IN mutants. Recent work has shown that IN can bind to gRNA, indicating that IN has a critical role in the maturation process and gRNA encapsidation. In accordance with previous findings, we hypothesize that IN acts as a physical bridge between the gRNA and assembling CA. Consistent with this hypothesis, preliminary data that we have collected suggests that IN binds to CA hexamers. Furthermore, cryoEM analyses indicate that IN likely binds to the C-terminal domain (CTD) of CA, causing a shift in CTD-CTD organization as the capsid assembles. Finally, RNPs purified from bona fide virions can be reencapsidated with recombinant CA that is assembled de novo. Further investigations proposed in this F32 application aim to uncover a detailed mechanistic binding scheme of IN and CA and will utilize complementary structural biology and biophysical techniques. The results are expected to further our understanding of the maturation process of HIV and possibly identify an attractive new target for antivirals.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY/ABSTRACT The global trend toward earlier pubertal development has extensive public health implications, particularly for low- and middle-income countries (LMICs) where age at menarche has been decreasing faster than in high- income countries (HICs). Additionally, in several LMICs, the timing of menarche has been found to vary by rural versus urban residence. As younger age at menarche is a risk factor for negative health outcomes like reproductive cancers, obesity, and cardiovascular disease, understanding the timing and determinants of pubertal development and menstrual characteristics may elucidate linkages between pubertal exposures and related health disparities. Therefore, we propose to characterize patterns in the timing, tempo, and determinants of pubertal development and menstrual characteristics in the Costa Rican study, Estudio de Comparacion de Una y Dos Dosis de Vacunas Contra el Virus de Papiloma Humano (ESCUDDO). We hypothesize that urban girls will experience earlier puberty, faster tempo, and shorter, regular menstrual cycles compared to their rural counterparts. We will leverage data from the full ESCUDDO cohort and the Immunogenicity Subcohort Group (ISG) which comprise of pubertal development and menstrual history from Costa Rican girls ages 12-21 and ages 12-14 at baseline, respectively. We will first evaluate if the timing and tempo of pubertal development and menstrual characteristics is earlier and faster in urban girls compared to rural girls (Aim 1). Using data from the full cohort (n=24,782), we will ascertain differences in pubertal development and menstrual characteristics across the rural-urban continuum in girls ages 12 to 21 using parametric survival models for pubertal timing and mixed-effects models for pubertal tempo (Aim 1a). Using longitudinal data from the ISG (n=1,408) we will update our estimates of pubertal timing and tempo (Aim 1b). We will also determine if associations between determinants of earlier and faster pubertal development and menstrual characteristics in predominantly HICs are also determinants in Costa Rica (Aim 2) by measuring the effect of early pubertal development predictors including weight, BMI, prenatal and neonatal history, smoking, and socioeconomic status on the timing and tempo of pubertal development and menstrual characteristics using parametric survival models. The proposed study will be the largest comprehensive assessment of pubertal development and menstrual characteristics in female adolescents to date and will help identify populations at high risk of earlier, faster pubertal development and shorter, regular menstrual cycles. The objectives outlined in this proposal will provide me with extensive experience in reproductive health research, while contributing to the large knowledge gap of pubertal development globally. Furthermore, the combination of rigorous training in advanced epidemiologic methods, experiential learning, and expert mentorship will ensure my transition to a successful physician-scientist.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Squamous cell carcinoma (SCC) is one of the most common cancers worldwide, with over one million deaths annually. While treatments such as chemotherapy and radiation are available, their severe side effects highlight the need for targeted therapies. SCCs from diverse tissues often share the overexpression of the transcription factor SOX2, which has been implicated in tumor initiation and maintenance. Despite its central role in SCC, therapeutic targeting of SOX2 remains challenging, necessitating a deeper understanding of its oncogenic transcriptional program and regulatory mechanisms. Our preliminary work demonstrates that SOX2 acts primarily as a transcriptional activator in SCC, binding promoters and distal enhancers to upregulate cancer- relevant genes via chromatin loops. Interestingly, many SOX2 binding sites are co-occupied by the oncogenic transcription factor TP63, a known master regulator in SCC. TP63 also functions as a pioneer factor, opening chromatin to facilitate binding by other factors including SOX2. Despite their established co-regulatory role, little is known about the mechanisms by which SOX2 and TP63 interact to regulate target gene expression in SCC. We hypothesize that SOX2 and TP63 co-occupy enhancers and upregulate target genes driving oncogenic pathways in SCC. This study aims to elucidate the functional regulatory role of SOX2 in SCC by addressing two key aims. First, characterizing the cancer-specific transcriptional program regulated by SOX2. Using an integrated genomics approach combining CRISPR-Cas9-mediated SOX2 knockout RNA-seq, SOX2 ChIP-seq, and H3K27ac HiChIP, we will identify direct SOX2 target genes. We will then prioritize genes relevant to human SCC using data from the Cancer Genome Atlas. We will evaluate the role of these prioritized target genes in cell proliferation and tumor growth using an in vitro competition assay and in vivo xenograft models. Second, we will explore the interplay between SOX2 and TP63 at shared enhancer binding sites. We hypothesize that TP63 facilitates chromatin accessibility, enabling SOX2 binding, and together, they regulate critical oncogenic pathways. This will be tested using CRISPR Cas9-based enhancer motif disruption, and RNA-seq and ATAC-seq following TP63 knockout to assess changes in chromatin accessibility and gene expression. This work will reveal SOX2 target genes, and how SOX2 along with TP63 drives SCC, providing insights into cancer-specific pathways that could serve as therapeutic targets. Furthermore, the insights gained may extend to other cancers where SOX2 activation is implicated, broadening the impact of this work.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY The prevalence of metabolic disease is growing rapidly, driven predominantly by increasing incidence of obesity and metabolic-associated steatohepatitis (MASH). However, there are very limited therapies for treating liver injury, placing these as a growing area of unmet clinical need. One therapeutic approach for metabolic diseases has been to seek human gene variants conferring susceptibility or protection. Unfortunately, several of the top genetic targets have phenotypes in model systems that do not replicate human pathology, have loss-of-function phenotypes associated with higher injury risk, or are associated with worsening cardiovascular risk, complicating the development of drugs. Here, we propose to study a gene, EF-hand domain family member D1 (EFHD1), whose decreased hepatic expression in genome-wide analyses has been associated with protection from liver injury in multiple, racially-diverse human populations. The EFHD1 protein is targeted to the mitochondrial outer membrane, including areas interacting with the endoplasmic reticulum, and possesses Ca2+-binding EF hands. We find that mice without EFHD1 (Efhd1-/-) have no obvious adverse phenotypes at baseline, but are protected against liver injury during a metabolic challenge, replicating the human phenotype. Therefore, inhibiting EFHD1 may be a promising therapy for liver injury. Moreover, a widespread phenomenon noted in many organs, including the liver, is that overnutrition causes excessive mitochondrial fission through the release of Ca2+ from the ER. However, the critical transducer coupling ER Ca2+ release to mitochondrial fission has not been identified, despite decades of study. Here, we hypothesize that EFHD1 is, in fact, that critical transducer, providing a mechanistic understanding for why inhibiting it may be potentially beneficial in MASH. Our proposal comprises three aims. First, we will define whether EFHD1 is the critical adaptor facilitating Ca2+-dependent mitochondrial remodeling. Second, we will determine if loss of EFHD1 leads to altered fatty acid oxidation. Finally, we will test whether loss of EFHD1 is protective against hepatocyte damage and subsequent inflammatory responses in mouse models of MASH and liver injury. Our collaborative team of investigators combines expertise in mitochondrial Ca2+ signaling, mouse and human studies of liver disease, and rigorous metabolic phenotyping, and is well-poised to tackle this project. In summary, the series of experiments detailed in this proposal will reveal the mechanism of EFHD1 action, and establish whether targeting this pathway may be protective against liver injury, as seen in human genetic analyses.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY: Living organisms have evolved the capacity to cope with exposures to chemically diverse foreign entities, called xenobiotics. For humans, xenobiotics are frequently encountered from diet, cosmetics, industrial or agricultural products, environmental pollutants, and therapeutic drugs. Nearly all therapeutic drugs entering the body encounter the xenobiotic detoxication system (XDS) which acts to detect, chemically modify, and eliminate xenobiotics from the body. Therefore, the XDS governs drug safety and efficacy. However, the dynamic nature of the XDS is rarely considered when prescribing. Xenobiotic-triggered upregulation of the XDS is called induction. Evidence from my work indicates induction might be common in medically complex children, adolescents, and young adult (CAYA) patients who experience polypharmacy. A state of polypharmacy is prone to precipitate induction as well as cause broad disruptions in drug pharmacokinetics (PK) which can result in drug failure or toxicity. My central hypothesis is that xenobiotic- induced changes in the XDS are common, largely unrecognized, and contribute to suboptimal therapeutic outcomes for medically complex CAYA experiencing polypharmacy. In a clinical setting, assessing the phenomenon of induction is challenging and therefore rarely performed. Nonetheless, determining the frequency, magnitude, and triggering exposures for induction among medically complex CAYA could provide numerous opportunities to improve therapeutics use and benefit patient outcomes. My translational clinical pharmacology research program seeks to better understand factors that influence drug safety and efficacy in medically complex CAYA who experience polypharmacy. To accomplish this, I built a unique pharmacology platform (“PKPD platform”) to obtain much needed standard-of-care (‘real-world’) population- specific PK data. We found that blood concentrations, spanning commonly to rarely prescribed drugs across different pharmacologic classes were consistently lower than expected for achieving therapeutic target ranges. As polypharmacy is a near universal feature for medically complex CAYA, we began to suspect induction was an unrecognized but prevalent factor affecting drug therapy. In the timeframe of this proposal, we will use our PKPD platform to investigate the rate and magnitude of induction as well as causes and consequences of induction among medically complex CAYA. Additionally, we will evaluate a newly discovered mechanism of drug-induced changes to epigenetic regulator activity and the consequences for XDS gene expression.

NIH Research Projects · FY 2025 · 2025-08

Project Abstract: HIV is a continual threat to public health. Despite advances to prevent infection and slow disease progression, no definitive cure exists for this deadly virus. Existing treatments for HIV target essential viral proteins and enzymes, such as protease, reverse transcriptase, integrase, and viral capsid interactions, all of which contribute to the successful delivery of an infectious virus to the next host cell. While the major roles of these enzymes are well understood, the mechanism of their incorporation during viral assembly as part of the Gag-Pol polyprotein has yet to be defined. Preliminary data from the Saffarian Lab shows a mix of mature and immature Virus-Like Particles (VLPs) from expression of an NL43 backbone with a Psi (Ѱ) packaging signal deletion, NL43(Ѱ: Δ(105-278)&Δ(301-332)), in HEK293 cells. The Psi packaging signal deletion reduces viral gRNA incorporation during assembly, but the effect of the deletion on maturation efficiency has not been defined. This finding highlights the need for further investigation of the mechanism of Gag-Pol incorporation and the possible role of viral gRNA in maturation efficiency. This proposal aims to test two hypothesis to address these unknowns: Hypothesis 1- An unknown Gag-Pol packaging mechanism is disrupted by removal of the psi packaging signal, (Δ(105-278)&Δ(301-332)), resulting in VLPs lacking sufficient Gag-Pol for maturation. Hypothesis 2- There is a role for gRNA in protease activation, and virions that did not mature did not have sufficient gRNA or cellular RNA’s incorporated. Aim 1 will investigate the mechanism of Gag-Pol incorporation by examining the fluorescence of labeled Gag-Pol proteins in purified HIV VLPs. Aim 2 will uncover the role of viral gRNA in virion maturation efficiency by leveraging lentiviral vector systems to selectively incorporate gRNA domains into minimally viable VLPs. These VLPs will be examined for maturation efficiency using advanced Cryo-Electron Tomography and subtomogram averaging. These experiments will provide training in advanced fluorescent light microscopy, Cryo-Electron Tomography, and complimentary biochemical assays. Additionally, exposition of the proposal findings will offer experience in oral and written scientific communication by attending and presenting at conferences and publishing in high quality journals. These skills will serve the fellow as an independent researcher following the conclusion of the fellowship. Finally, the findings of this proposal stand to elucidate essential HIV maturation mechanisms, opening avenues for drug targets and advanced treatment for HIV.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Primary graft dysfunction (PGD) is a form of acute lung injury that occurs in the first 72 hours after lung transplantation. PGD is the leading cause of morbidity and mortality in the first 30 days and is strongly associated with an increased risk of chronic rejection and death beyond the first year after transplantation. Donor, recipient, and surgical risk factors for PGD have been identified, and clinicians make decisions about donor offer acceptance based on these risk factors to avoid severe PGD. However, how different donor and recipient risk factors interact to impact the risk of PGD is unclear. We previously built a machine learning (ML) algorithm to predict the risk of severe PGD using clinical data available at the time of donor offer. However, this model was based on single-center data, and its performance could be improved by capturing additional key parameters using a multi-center cohort. Furthermore, the model did not include donor lung CT images through imaging is a cornerstone of clinical decision-making. We hypothesize that donor lung CT images encode valuable information that could be used to estimate the risk of severe PGD and that better risk estimates could be achieved by fusing donor and recipient clinical data with donor CT imaging data. To test this hypothesis, we will perform an observational study at five lung transplant centers in the US. We will use a retrospective cohort of bilateral lung transplant recipients from the sites to build ML models to predict severe PGD, and concurrently, enroll lung transplant recipients in a prospective cohort, which will serve for model validation. To determine the characteristics of donor CT images associated with severe PGD, we will use the attention-based multiple instance learning (MIL) method. We will divide CT scans into 3D patches of 32×32×32 voxels per patch, and severe PGD will serve as the pseudo-label for all 3D patches. Both the original CT scans and the augmented sub-volumes will be processed through separate Swin Transformer encoders and combined into three “heads” used to compute the inpaint loss, contrastive loss, and bounding box detection loss. Gradient-weighted class activation mapping (Grad-CAM) will be applied to the attention scores of the 3D patches to visualize prioritized lung regions. Attention masks generated by the MIL model will be examined by a thoracic radiology expert to identify and list imaging biomarkers predictive of severe PGD. We will then integrate the clinical and imaging data using a multimodal learning strategy. The clinical and MIL-identified featuresets will be aligned using separate Multi-Layer Perceptron (MLP) layers before concatenation and passed through another series of MLP layers. To avoid overfitting, clinical and MIL-identified features will be combined in an element-wise fashion with their independent feature vectors before being passed through the final MLP layers. This project will develop a multi-dimensional clinical decision-making tool to predict the risk of severe PGD. This can enhance donor- recipient matching and clinical preparedness for immediate post-transplant care to improve early outcomes after lung transplantation.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract The goal of this application is to support Alan Morris, MS, as a Research Software Engineer at the Scientific Computing and Imaging Institute at the University of Utah. He works primarily on research software supported by NIH grants under the direction of Drs. Shireen Elhabian and Andrew Anderson. The primary focus of his work is the development of software for medical image and shape analysis and in supporting the research of the faculty and students at the SCI Institute. This Research Specialist Award would allow 3 years of funding to maintain protected time for Mr. Morris to work on supporting NIH-related research software projects with the particular focus of enhancing them with robust machine learning support and infrastructure. The results of this work will be disseminated through open-source software releases, peer-reviewed journal manuscripts, and presentations at local and national meetings.

NIH Research Projects · FY 2025 · 2025-08

Inappropriate activation of the Hedgehog (HH) pathway drives basal cell carcinoma (BCC) of the skin, the most common cancer in the United States with greater than 4 million cases annually. Existing drugs target oncogenic HH signaling by inhibiting the key pathway regulator SMOOTHENED (SMO). Tumors initially respond to SMO inhibitors, but these molecules ultimately lose effectiveness, largely due to secondary mutations within SMO that render it drug-resistant. To elicit expression of oncogenic target genes involved in tumorigenesis, SMO signals intracellularly to GLI transcription factors, but for decades the underlying signaling mechanism remained obscure. We recently identified a key aspect of SMO-GLI communication that enables us to propose new therapeutic strategies to minimize resistance issues. Working in cultured cells and embryos, we discovered that the active, agonist-bound form of SMO undergoes phosphorylation by the kinase GRK2, enabling SMO to directly bind the PKA catalytic subunit, inhibit its enzymatic activity, and thereby prevent PKA from phosphorylating and inactivating GLI. We hypothesize that this novel SMO-GRK2-PKA signaling pathway transmits oncogenic HH signals from the cell surface to the nucleus during BCC tumorigenesis. As such, GRK2 represents a promising therapeutic target for both primary and SMO inhibitor-resistant BCC. The goal of our MPI R01 grant is to define the role of GRK2 in mediating oncogenic SMO-GLI communication in BCC, elucidate the underlying mechanism of phosphorylation, and to evaluate GRK2 inhibition as a BCC therapeutic strategy. Using cultured cell and in vivo BCC models, we will determine whether SMO undergoes GRK2 phosphorylation in primary cilia and probe the functional significance of these phosphorylation events to oncogenic GLI activation and BCC tumorigenesis. We will also block GRK2 activity using either genetic or pharmacological approaches, the latter capitalizing on selective, high-affinity, bioavailable GRK2 inhibitors that have already been developed and utilized in cardiovascular disease models. Lastly, we will uncover the biochemical and cell biological mechanism by which GRK2 recognizes the SMO active conformation at the cell membrane, which is its essential role in HH signal transduction, and capture a cryoEM structure of SMO-GRK2 or SMO-GRK2-Gβγ complex to understand this process at atomic resolution. To carry out our studies, we have assembled a team of experts in biochemical / cell biological mechanisms of HH signal transduction (Myers), BCC tumorigenesis (Atwood), and GRK2 structure / pharmacology (Tesmer). We expect these studies to define an essential and unique role for GRK2 phosphorylation of SMO in transmitting oncogenic HH signals from the cell surface to the nucleus, and for GRK2 inhibition, either alone or as a combination therapy with FDA-approved SMO inhibitors, to block tumorigenesis and forestall resistance more effectively than existing standard methods of care. Overall, our work will provide valuable insights into the mechanism of BCC tumorigenesis and validate a novel BCC therapeutic strategy, with the potential to transform treatment paradigms for many malignancies driven by ectopic SMO or GRK2 activity.

NIH Research Projects · FY 2025 · 2025-08

PROJECT ABSTRACT Eosinophilic Esophagitis (EoE) is a morbid, highly inflammatory condition of the esophagus, increasingly diagnosed in both children and adults, resulting in substantial healthcare burden for patients and payors. Due to the need for invasive upper endoscopies for diagnosis, the disease is not identified for years after the onset of symptoms. Yet, identifying and controlling the disease early in its development would provide a means to personalize therapy and reduce complications. Our long-term goal is to detect and characterize the early epithelial signals which precede the development of persistent, chronic EoE, identifying biomarkers of early disease, in order to reduce the burden of this morbid, inflammatory process. The overall objective for this application is to characterize the molecular pathways responsible for disease initiation, differentiating them from those that are responsible for the maintenance and persistence of inflammation over years. Our central hypothesis is that the initiation of EoE involves specific reversible (preventable) epithelial signals which propagate and perpetuate a robust, complex, inflammatory reaction leading to diffuse esophageal inflammation, fibrosis, narrowing, and life-altering symptoms of choking, food impactions, and food/social aversion. Our rationale stems from our discovery that specific pathways are differentially regulated within esophageal tissues during food elimination/reintroduction diets. The central hypothesis will be tested by pursuing two specific aims: (1) to differentiate RNA signatures responsible for the initiation of EoE (acute EoE) from those involved in persistent, longstanding inflammation (chronic EoE);and (2) to identify the specific epithelial and immune pathways which precede the onset of robust, diffuse esophageal eosinophilia. Under the Aim 1, the RNA signatures from paired biopsies taken prior to food elimination (“chronic” EoE) and after recurrence of EoE during reintroduction of trigger foods (“acute” disease phase) within patients will be compared to differentiate the processes involved in disease initiation and disease persistence. For the Aim 2, tissues from patients who resolved their EoE with food elimination diet and subsequently demonstrated “patchy” recurrence of disease upon trigger food reintroduction (i.e. obvious eosinophilic inflammation and endoscopic signs of EoE interspersed with normal-appearing tissue) will undergo spatial transcriptome sequencing to localize and decipher the cellular contributions involved in disease recurrence. The proposed research is innovative in its study of human samples to thoroughly categorize early changes involved in disease initiation. This study of early transcriptional signals capitalizes on biopsies performed during food elimination/reintroduction diets which remove and then expose patients to the antigenic stimulus – essentially recreating a scenario which mimics disease genesis. The significance of the research lies in the identification of new biomarkers and eventual interventions/therapies for early stages of EoE. Collectively, our observations should inform future investigation into early markers of disease and the development of pre-emptive therapies to prevent disease progression.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract SCIRun, is an open-source problem solving environment that was developed in previous NIH-funded projects to enable all aspects of bioelectric field simulation, with a special emphasis on applications to neuromodulation and stimulation of the brain. Many advancements in transcranial (TMS, tCS) and invasive stimulation (DBS, sEEG, ECoG) have relied on computational simulation and optimization methods and many have used SCIRun to perform these simulations. However, SCIRun’s capabilities have fallen behind as stimulation methods have expanded (e.g., more complex stimulation electrodes with different sizes and numbers of electrodes). Moreover, any software product, especially with the complexity of SCIRun, also requires constant updates to remain com- patible with emerging operating systems and support libraries. We propose major upgrades and expansions of SCIRun through technological, infrastructure, interface, and training advancements. These improvements will make the package more efficient, flexible, and comprehensive. We will strongly broaden its use cases for scientists by developing new stimulation methods that may improve treatment outcomes and reduce costs for a range of pathologies. These improvements are also specifically aimed at making SCIRun more accessible to clinicians who can use our tools for individualized treatment planning and increased patient understanding. Of special interest will be applications in neuromodulation, a domain with vibrant growth in the exploration of novel electrode configurations and target interventions. To support these specific innovations will require improve- ments to the general software environment to reflect contemporary computing resources. Such changes in the research computing ecosystem provide opportunities for pipeline integration, distribution, and democratization of computing access. Changes that we wish to leverage include the explosion of Python as a development environment, which has seen considerable investment in infrastructure, support, capabilities, and distribution that remain open source and accessible. Similarly, improvements in cloud computing, web applications, and deployment technology have made these avenues more accessible. In addition to improving capabilities, inter- operability, and distribution, leveraging these open-source tools and ecosystems will connect us to active user communities, providing further opportunities for exposure, support, and sustainable benefits.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY We often think of action potentials as a way for cells to quickly send signals over long distances and release substances like neurotransmitters. This perspective overlooks the crucial role of electrical signals in triggering intracellular biochemical processes, including gene expression. Our research has identified that endoplasmic reticulum (ER)-plasma membrane (PM) junctions in the neuronal soma act as specialized sites for this electrical- biochemical coupling. These ER-PM junctions are molecularly complex, exhibiting diverse architectures, protein compositions, and distributions, indicating that they serve distinct cellular functions. Our lab's long-term goal is to understand how electrical signals are translated into specific biochemical responses, such as activating gene expression programs. We propose that ER-PM junctions are central to translating membrane depolarizations into intracellular signaling events, and our work is focused on discovering the mechanisms underlying their assembly and function. Building on our previous work in defining the proteome and physiological roles of these junctions in mammalian neurons, our five-year goal is to investigate the regulatory mechanisms governing their organization, with a particular emphasis on cAMP-dependent protein kinase (PKA) signaling. First, we aim to define the signaling interactions at PKA-containing ER-PM junctions, focusing on how distinct A-kinase anchoring proteins (AKAPs) recruit additional scaffolding proteins and shape the composition of these signaling complexes. We will use genetic and biochemical approaches to map enzyme substrates targeted to these sites and identify how these enzymes influence somatic signaling pathways. Secondly, we will examine how AKAP- enzyme signaling complexes are structurally organized, especially how conserved, disease-linked domains within AKAPs direct the formation of specific complexes at ER-PM junctions. A key aspect of our research will be to determine how PKA’s kinase activity versus its role in organizing ER-PM junctions affects signaling outcomes. By identifying the molecular components and pathways involved, this research aims to clarify the roles of ER-PM junctions in neurons and uncover principles applicable to similar structures in other cell types. Such knowledge will improve our understanding of how electrically excitable cells fine-tune their signaling systems to regulate processes like gene expression.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Up to 1 in 5 women undergo surgical treatment for pelvic organ prolapse (POP) yet we know little about its pathogenesis; thus, women receive interventions for POP only once they have reached end-stage disease. Identification of early markers of POP (precursor phenotypes) is needed to identify women at risk of end-stage POP and to develop effective preventative strategies, which currently do not exist. Our long-term goal is to change the status quo from treatment of end-stage POP to prevention of early POP through screening and early detection. Well-established risk factors for POP include aging, VD, and family history (FH). The Lifespan Model posits that maximal pelvic floor functional reserve (PF reserve), or its capacity to function optimally at full growth based on each person’s unique structural properties, is largely predetermined. PF reserve declines throughout life, due primarily to VD and aging. End-stage POP develops once PF reserve is impaired below a specific threshold and those who begin with a lower PF reserve will reach this threshold sooner. Women who undergo POP surgery before they reach 45 years are 5-fold more likely to have a FH of POP, supporting the concept that PF reserve is heritable. Childbirth induced levator ani muscle (LAM) avulsion, where the pelvic floor muscle is torn, is a major risk factor for POP, but is only present in ~50% of POP cases. An understanding of PF reserve will fill in critical knowledge gaps about the pathogenesis of POP, why effects of VD and aging differ amongst women according to their FH risk, and ultimately will provide avenues to identify prevention and early therapeutic strategies based on phenotypes of PF reserve. Using MRI, clinical measures, and RNA sequencing of the LAM, we plan to quantify structural, functional, and cellular & molecular domains of PF reserve in women without LAM avulsion and prior to the onset of age- related changes. Four groups of women, 18-30 yo, will be selected based on FH and VD risk. The objective of Aim 1 is to quantify the effect of FH risk on baseline PF reserve prior to VD. The objective of Aim 2 is to quantify the effect of FH risk on PF reserve after the first VD. Using Aims 1&2 data, we will determine, for the first time in humans, correlations between primary cellular/molecular pathways and structural/functional pelvic floor parameters. In Aim 3, we will apply these PF reserve results to a validated biomechanical platform (POP- SIM) to quantify PF reserve and identify parameters with the greatest impact on POP development. Our overall objective is to test the hypothesis that FH risk, defined as ≥1 first-degree relative with POP, impairs PF reserve and that VD increases the magnitude of that effect in the absence of well-established risk factors of LAM avulsion and aging. At the end of this project, we expect to have identified POP precursor phenotypes that can be validated in future studies and used to develop a new domain of research in the area of POP prevention.

NIH Research Projects · FY 2025 · 2025-08

Childhood asthma is a leading cause of chronic disease and hospitalization in children. Viral infections within the first year or two of life are strongly associated with childhood asthma risk, but the immune mechanisms mediating this association are poorly understood. The long-term goal of this proposal is to define how early-life inflammation determines asthma susceptibility by reprogramming the fate and function of type 2 innate lymphoid cells from hematopoietic progenitors during early life. This proposal builds on preliminary data linking Type I interferon-mediated inflammation induced by maternal immune activation with poly(I:C) and associated with common viral infections with a hyperactivated Type 2 innate lymphoid cell (ILC2) response in the postnatal lung that drives asthma susceptibility in mice. ILC2s are strongly implicated in asthma pathogenesis in mouse models and have also been identified in human asthma, but their role as mediators of asthma susceptibility is undefined. The overall objectives of this proposal are to define the cellular and molecular mechanisms by which viral-induced inflammation initiates a hyperactivated ILC2 phenotype at the progenitor level and determine the requirement and sufficiency for hyperactivated ILC2s in remodeling lung immunity and susceptibility to asthma during early life. We will also examine hyperactivated ILC2s in human development in response to infection as a putative biomarker for asthma susceptibility. The central hypothesis of this proposal is that early-life infections promote lung dysfunction and asthma susceptibility by programming hematopoietic progenitors to produce hyperactive tissue-resident ILC2s that shape lung immunity. We will test this hypothesis in three specific aims. In Aim 1, we will define the cellular and molecular mechanisms underlying the persistent hyperactivated function of ILC2s in response to early-life inflammation by performing progenitor fate-mapping and orthogonal transcriptomic and epigenomic analyses, In Aim 2, we will test whether hyperactivated ILC2s in the postnatal lung are necessary and sufficient to induce lung immune remodeling and lung dysfunction using a combination of specific gene deletion models and transplantation approaches. In Aim 3, we will perform phenotypic and functional investigation of human umbilical cord blood ILC2s from normal pregnancies and pregnancies affected by chorioamnionitis to gage the response of human ILC2s to early-life infection. We will also leverage comparisons to our own single-cell transcriptomic datasets in mice to define transcriptional regulators of the human ILC2 response. The concept of this proposal is innovative in defining perturbation of “layered immune development” of ILC2s as a novel pathogenic driver of asthma susceptibility, and innovative in approach by using overlapping inducible fate-mapping and genetic deletion models in parallel with single-cell transcriptomics, ATAC-seq, and adoptive transfer assays. The proposed research is significant because it will address gaps in understanding asthma pathogenesis and will pave the way for the identification of new therapeutic targets for asthma treatment and prevention.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT CD4+ T cells mediate protection from influenza infection in mice and humans and represent a potential target of vaccination. While influenza-specific T cells may be generated via immunization or viral infection, how the nature of initial antigen exposure shapes the formation, persistence and function of the subsequent CD4+ T cell response to influenza infection is not fully understood. The long-term objective of our research is to determine the TCR-dependent and epigenetic mechanisms that drive secondary CD4+ T cells responding to influenza infection. We recently published a collaborative study showing that CD4+ T cell responses to influenza are shaped by prior antigen exposure. We adopted a sequential heterologous immunization approach in which CD4+ memory T cells generated via either infection or protein immunization are rechallenged with influenza virus. Both infection- and immunization-induced CD4+ memory T cells undergo extensive secondary expansion following influenza infection, leading to the formation of long-lived and abundant secondary CD4+ memory T cells, both in circulation and in the lung, as well as enhanced germinal center responses. The function of the secondary response in the lung is guided by the primary challenge. Primary activation of CD4+ T cells via protein immunization biases the lung CD4 secondary response to influenza towards Tfh function, whereas primary CD4+ T cell activation via infection biases the lung CD4 secondary response to influenza towards Th1 function. Our preliminary data indicate that CD4 effector T cell responses in the lung are also biased towards high TCR signaling and high affinity, as compared to the draining lymph node, suggesting that strong TCR signals may be required to sustain effector responses and drive the formation of large numbers of secondary CD4+ memory T cells. Additionally, we previously reported that the functional differentiation of CD4+ T cells in response to acute infection is controlled epigenetically by acquisition of DNA methylation programing, with the opposing effects of the DNA methyltransferase DNMT3A and the methylcytosine dioxygenase TET2 determining differentiation outcome and memory T cell plasticity. Therefore, we will test the overall hypothesis that TCR signal strength and epigenetic DNA methylation programming guide the function and long-term memory of the secondary CD4 T cell response to influenza infection. Our aims are to: 1) Determine the relationship of TCR signal strength and TCR antigen-binding kinetics, including affinity and bond lifetime under force, to secondary CD4+ T cell differentiation in the lung; and 2) Define the role of TET2- and DNMT3A-dependent epigenetic programming established during primary T cell activation in driving secondary effector function in response to influenza challenge.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY Strikingly, cancer patients with a variant form of HER2 have a median progression free survival (PFS) of 4.9 months – a stark contrast to generic reports that HER2 targeted therapy gives 90% of patients a PFS of 5+ years. Most resistance is due to HER2 variants – structural mutants that prevent drug binding or drastically decrease the internalization of HER2. Accordingly, most standard and emerging treatments fail for these patients. There is an immediate need for therapeutic approaches that can be specifically designed to treat HER2 variants. Antibody-directed enzyme prodrug therapy (ADEPT), a strategy of conjugating prodrug-activating enzymes to antibodies that target cell surface receptors, nearly reached clinical success. It was limited, in large part, by premature prodrug activation in circulation. We propose to re-engineer the ADEPT strategy, addressing previous shortcomings, to tackle the pressing clinical need resulting from HER2 variants. In our strategy, Complementation Dependent Enzyme Prodrug Therapy (CoDEPT), split-enzyme fragments are fused to two separate antibodies against HER2. The binding of these antibodies to HER2, at distinct epitopes, brings the split- enzyme fragments into proximity, facilitating reconstitution of the active enzyme. The enzyme subsequently activates prodrug at the tumor and should thereby drastically reduce off-site activation and related toxicity. We hypothesize that CoDEPT constructs can reduce tumor growth in a murine breast cancer model. In Aim 1 we will engineer and characterize a library of split-β-lac constructs targeting HER2 variants and select lead CoDEPT constructs. In Aim 2, we will determine kinetic parameters and concentration thresholds that will establish foundational requirements for CoDEPT prodrug delivery and activation. In Aim 3, we will evaluate therapeutic efficacy of select CoDEPT constructs in vitro and in vivo.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The IGF2BP family of RNA binding proteins were recently shown to interact with multiple ASD-associated transcripts and proteins, suggesting it is part of a convergent ASD pathway. However, the fundamental role of IGF2BPs in brain development is poorly defined and is the focus of this proposal. Here we report the discovery of new missense variants for IGF2BP3 found in patients with neurodevelopmental disorders, and phenotype modeling in Drosophila is used to study their function. Loss of the fly IGF2BP3 orthologue Imp from post-mitotic neurons causes microcephaly, and these phenotypes are rescued by wildtype human IGF2BP3 but not patient- associated variants. Imp is known to promote stem cell division, so the finding that Imp has essential functions in, and causes microcephaly when depleted from, non-dividing post-mitotic neurons is novel and surprising yet critical to our understanding of human disease throughout life. IGF2BP1-3 and Imp are RNA-binding proteins that regulate many mRNA targets by modifying stability, transport, splicing, or protein translation. Sap47 (Synapse associate protein 47) is one such target as loss of Sap47 from postmitotic neurons causes microcephaly in flies just like loss of Imp. Sap47 localizes to synapses, but its function in neurons remains largely unknown. Moreover, its mammalian orthologue SYAP1 is a strong ASD risk factor but its function in the mammalian brain is also completely unknown. Imp and Sap47 adversely affect both neuron cell survival and morphology, which likely contribute to microcephaly and a miswired brain characteristic of multi-factorial neurodevelopmental disorders. This proposal will test the hypothesis that IGF2BP3/Imp stabilizes Sap47/SYAP1 mRNA in neurons to promote neuronal targeting and survival and thereby ensure proper brain development. A combination of fly and mouse models as well as human gene variants will be used to investigate the conservation of protein function. First, how Imp and Sap47 loss results in defects in neuron outgrowth, targeting, and survival defects will be investigated using highly characterized fly visual system neurons and mouse hippocampal cultures. The molecular mechanism by which Imp/IGF2BP1-3 regulates Sap47/Syap1 mRNA in neurons will be determined by testing Imp’s role in mRNA stability. Pulse-chase in vivo 5-ethynyluridine (EU) incorporation assays will test Sap47 mRNA stability with and without Imp. IGF2BP3 and patient variants will also be tested. Finally, the mechanism by which Sap47 ensures proper brain development and function will be elucidated using domain analysis. In addition, work here will determine whether loss of putative Sap47 protein interactors cause brain volume phenotypes or modify Imp and Sap47 phenotypes in neurons. Mammalian cell culture will be used to test molecular and biochemical conservation of identified interactions.

NIH Research Projects · FY 2025 · 2025-08

Project Summary CD4+ helper T cells provide critical effector functions to combat viral diseases and are a key cell type for vaccine- induced immunological protection against viruses. During response to chronic viral infections, stimulation by persistent antigen and inflammatory cytokines can induce dysfunction that limits helper T cell function that contributes to subsequent viral persistence and chronic viral diseases. While such dysfunction is well-described for exhausted CD8+ T cells generated by chronic antigen stimulation, the mechanistic basis for dysfunctional CD4+ T cells generated during chronic infection are poorly understood. We hypothesize that the dysfunctional state of virus-specific CD4+ T cells during chronic infection is programed in part through the acquisition of epigenetic programing by changes in DNA methylation at key genes that regulate T cell functions such as cell proliferation, survival, and effector functions. This hypothesis is based on our key preliminary data from our studies of virus-specific CD4+ T cells that are deficient in Tet2 and Dnmt3a, enzymes that regulate active demethylation and de novo methylation of CpG dinucleotides of DNA, respectively. Using a highly novel experimental model to study cell-intrinsic programing of virus-specific CD4+ T cells, we have found that the combined deficiency of Tet2 and Dnmt3a results in CD4+ T cells with a remarkably unique capacity to maintain proliferative, survival, and functional capacities during chronic viral infection. The three specific aims of our proposal will define how chronic antigen stimulation contributes to Tet2-mediated active demethylation and Dnmt3a-mediated de novo methylation programing that drives CD4+ T cell dysfunction. We will use highly innovative conditional knockout in vivo virus-specific CD4+ T cell models, combined with whole genome DNA methylation sequencing to identify epigenomic programing and associated transcription factor pathways that contribute to impaired proliferation, survival, and effector functions of chronically-stimulated CD4+ T cells. Finally, we will determine whether disruption of such epigenetic programing results in CD4+ T cells capable of improved help for exhausted CD8+ T cells and improved viral control and clearance of chronic viral infections. Together, these studies will reveal critical mechanistic insights into CD4+ T cell programing that could be targeted for improving strategies to prevent and treat chronic viral infections.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Stroke is a leading cause of long-term disability worldwide and 80% of all stroke survivors experience upper-limb hemiparesis (i.e., weakening of motor control in the hand and arm on one side)1. Upwards of 50% of individuals with upper-limb hemiparesis also experience tactile sensory deficits in the hand2–4. However, tactile sensory deficits and the fine sensorimotor function that stems from tactile sensory feedback are frequently overlooked in today’s rehabilitation programs5,6. This is despite research indicating that the integration of both sensory and motor training can be more effective than conventional approaches which focus primarily on motor7. This neglect of fine sensorimotor function can be explained in part due to a lack of standardized assessments, rehabilitation tools, and validated performance metrics5,8. Given the importance of fine sensorimotor hand function in daily life, there exists a critical need for a standardized device capable of both assessing and rehabilitating fine sensorimotor hand function. Practicing grip force regulation, as one would do when handling a fragile object, presents a novel and unique method to both assess and rehabilitate fine sensorimotor hand function, as grip force regulation relies heavily on tactile feedback and fine motor control9,10. To assess and rehabilitate fine sensorimotor hand function after stroke, we are developing the Electronic Grip Gauge (EGG), a handheld wireless device that can mimic a fragile object. The EGG consists of a 3D-printed block embedded with sensors that track applied grip force, load force, acceleration, and relative object position. Stroke patients attempt to transfer the EGG over a barrier as quickly as possible while also regulating their grip force to avoid “breaking” the object. If applied grip force exceeds a set threshold, then an audible “break” sound plays. As a fine sensorimotor assessment, one must rely on their underlying tactile feedback to monitor and regulate their fine motor execution during transfers. The objective of this proposal is to validate the EGG as a diagnostic and rehabilitative tool for fine sensorimotor function, and to demonstrate performance marker reliability in stroke populations. Our central hypothesis is that an individual’s ability to rapidly transfer fragile objects without breaking them can provide quantifiable performance metrics for fine sensorimotor function. This application outlines a rigorous scientific and clinical training plan at the University of Utah Neurorobotics Lab which is housed within the Craig H. Neilsen Rehabilitation Hospital. Mentorship from leaders in stroke rehabilitation and clinical trials research will be combined with longitudinal clinical experiences in the Department of Physical Medicine and Rehabilitation. These activities will enable the applicant to become a successful physician-scientist in physical medicine and rehabilitation with a focus on running a future lab dedicated to the development and translation of novel rehabilitation devices.

NIH Research Projects · FY 2026 · 2025-08

ABSTRACT Medulloblastoma (MB) is the most common malignant pediatric brain cancer. A third of MB, called Group 2 or SHH MB, arises from the overactivation of a highly conserved cell-cell communication circuit called the Hedgehog (Hh) pathway. Several FDA-approved drugs target oncogenic Hh signaling by inhibiting SMOOTHENED (SMO), the key switch that regulates Hh pathway activity. Group 2 MB initially respond to SMO inhibitors, but these molecules ultimately lose their effectiveness due to secondary mutations within SMO. Clearly, new strategies are needed to prevent or overcome SMO-mediated drug resistance to improve patient outcomes. In this proposal, our goal is to harness new biochemical discoveries of SMO activation from our labs that identified 1) a new ligand-binding pocket deep within the SMO seven-transmembrane (7TM) region that explains how resistance emerges in SMO and suggests strategies to minimize it in the future and 2) G protein- coupled receptor kinases 2 and 3 (GRK2/3) as essential for SMO to activate downstream glioma-associated (GLI) transcription factors and stimulate expression of pro-oncogenic genes, ultimately driving tumorigenesis. Our findings immediately suggest two clear therapeutic strategies to combat chemotherapy resistance in SMO- driven malignancies. First, we hypothesize that a distinct class of SMO inhibitors which bind deep within the 7TM pocket and overlap extensively with the endogenous cholesterol ligand will be less prone to resistance than existing agents. Second, we hypothesize that GRK2/3 inhibitors will block Hh signaling arising from all existing oncogenic or therapy-resistant forms of SMO. Furthermore, we expect that combining SMO and GRK2/3 inhibitors will be even more effective than either inhibitor alone. We provide significant published and preliminary data to support these hypotheses and now propose to evaluate the extent to which SMO deep-pocket inhibitors and/or GRK2/3 inhibitors generate less tumor resistance than conventional therapies by rigorously evaluating these inhibitors using multiple assay platforms and species (fish, mice, human) of Group 2 MB. We will generate highly scalable in vivo brain tumor models in zebrafish to rapidly interrogate the impact of SMO and GRK2/3 inhibitors and leverage this knowledge to selectively test single or combination strategies as effective treatments that minimize drug resistance in genetically engineered mouse and human cell transplant models of Group 2 MB. We have assembled a team of experts covering the spectrum from biochemical/structural mechanisms of Hh pathway activation, animal models of MB and pre-clinical human cell-based in vivo models. Success in these studies will establish deep-pocket SMO and GRK2/3 inhibitors, alone or in combination, as new viable therapeutic strategies to treat Group 2 MB that forestalls or overcomes resistance more effectively than conventional SMO inhibitors. These studies have the potential to transform current treatment paradigms for Group 2 MB and other Hh-driven brain tumors and provide justification to design future clinical trials in children.