Utah State Higher Education System--University Of Utah

NIH Research Projects · FY 2025 · 2025-08

This proposal describes a 5-year intensive mentoring program and research plan with the goal of developing the principal investigator into an independent, patient-oriented clinical researcher focusing on the pathophysiology of cardiogenic shock (CS) and its overlap with the cardiorenal syndrome. The applicant is an advanced heart failure cardiologist with formal training through a Master of Science in Clinical Epidemiology and is currently an Assistant Professor at the University of Utah. CS is a life-threatening complication of myocardial infarction or heart failure with a rising incidence but persistent 30-40% in-hospital mortality. Acute kidney injury (AKI) and pathophysiologic vasodilation are strong and consistent risk factors for CS death, but the mechanisms linking these risk factors with CS death and with each other are poorly understood. The central hypothesis of this proposal is that Angiotensin II (Ang II) becomes deficient in a subset of patients with CS resulting in AKI, vasodilation, and death. This is supported by our preliminary data showing diminished Ang II levels in CS despite excess renin and Ang I. A similar mechanism has been observed in sepsis, leading to approval of synthetic Ang II as a treatment for vasodilatory shock, but its role in mixed CS and vasodilatory shock is unknown. This model challenges the paradigm that RAAS activity is always increased and always detrimental in heart failure patients, even when it is required for the regulation of peripheral vascular resistance and maintenance of glomerular filtration in states of reduced renal blood flow, and it provides a novel and potentially druggable pathway for the treatment of CS. The goal of this K23 is to initiate a mentored research project and training plan to a) establish a body of evidence for Ang II deficiency as a contributor to CS death, vasodilation, and cardiorenal pathophysiology and b) obtain training, education, experience, and data needed to transition to independence as a patient-oriented clinical researcher. The aims of this proposal are to i) use advanced longitudinal data analysis techniques to evaluate the dynamic association of pathophysiologic vasodilation in CS with AKI and death, ii) show, in a prospective CS cohort study, that Ang II deficiency is associated with CS vasodilation independent of the systemic inflammatory response, and iii) estimate the causal effects of vasopressors on AKI and death in CS after accounting for time-varying confounding from circulating Ang II levels using marginal structural models. A comprehensive training plan is developed to integrate with these aims to 1) master new skills in causal inference and longitudinal data analysis, 2) gain robust experience leading prospective multicenter research in a critical care setting, and 3) learn to analyze cardiorenal outcomes and measure and interpret key cardiorenal biomarkers, under the guidance of mentors expert in these methods. This K23 award will provide the resources necessary to support the Candidate’s transition to an independent research career in an area of growing public health importance.

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 · 2025-07

PROJECT SUMMARY Autoimmune diseases (ADs) arise due to a misguided immune response, where autoreactive lymphocytes launch attacks on the body's own tissues. Many AD patients, such as those of type-1 diabetes and multiple sclerosis, still suffer from ineffective treatments that fail to halt disease progression. Moreover, current AD treatments often employ an indiscriminate approach by suppressing both activated and naive lymphocytes. This leads to long-term lymphopenia, broad immunosuppression, and increased susceptibility to infections and malignancies. A promising alternative involves the targeted depletion of lymphocytes expressing programmed death-1 (PD-1), offering a novel solution to halt auto-attacks while avoiding long-term lymphopenia or immunosuppression. Additionally, it spares naive lymphocytes that are PD-1 negative, preserving the lymphocyte repertoire and allowing patients to quickly regain full immune protection once the depleting agents are cleared. Additionally, the depletion will spare naive lymphocytes that are PD-1 negative. The preservation of naive cells and lymphocyte repertoires will quickly re-gear treatment subjects with full immune protection after the cell depleting agents are cleared. Importantly, the use of an immunotoxin to deplete PD-1+ cells has confirmed these advantages. Together, these findings support the development of clinically viable agents for targeted lymphocyte depletion. In pursuit of this goal, we focused on a class of antibodies called Bispecific Killer cell Engagers (BiKEs). BiKEs are designed to enhance the interaction of natural killer (NK) cells and target cells by simultaneously binding target cell antigens and NK cell receptors. BiKEs have shown considerable success in clinical and preclinical settings, demonstrating superior efficacy in eliminating target cells compared to conventional depleting antibodies. Notably, BiKEs excel in eliminating target cells with modest antigen expression that is similar to the case of PD-1. Our innovative approach involves developing and utilizing anti-PD-1 BiKEs (PD-1 BiKEs) designed to bridge NK cells to PD-1+ cells, activate NK cells, and redirect their cytotoxicity towards PD-1+ cells. We hypothesize that PD-1 BiKEs have the potential to efficiently deplete primary PD-1+ cells and, consequently, mitigate disease progression in mouse models of ADs. We have successfully produced and characterized an anti-human PD-1 BiKE that targets the activating NK cell receptor CD16. This BiKE effectively initiates NK cell- mediated depletion of PD-1+ cells. These preliminary results underscore the rationale and feasibility of using BiKEs to deplete PD-1+ cells and alleviate ADs. To test our hypothesis, we will undertake the following two aims: Aim 1: Characterization and Functional Comparison of Mouse PD-1 BiKEs. Aim 2: Establishment of the efficacy and safety advantages of mouse PD-1 BiKEs.

NIH Research Projects · FY 2025 · 2025-07

Abstract Cerebral malaria is a severe complication of Plasmodium infection that can result in death. The main pathogenic effector cells are CD8 T cells that produce IFN-g; however, it is known that invariant natural killer T (iNKT) cells are needed in the Plasmodium berghei ANKA experimental cerebral malaria (ECM) model. The overarching hypothesis of this proposal is that iNKT cells provide the initial cytokines that drive pathogenic IFN-g producing anti-Plasmodium CD8 T cells. iNKT cells are a unique group of innate T cells that recognize glycolipid antigens and have the ability to produce cytokines rapidly following TCR stimulation. Although iNKT cells differ from conventional T cells in the types of antigens they recognize, iNKT cells are similar in that they produce effector cytokines such as IFN-g, IL-4, and IL-17 that can be characterized as iNKT1, iNKT2, and iNKT17 subsets respectively. These cells play important roles in the control of cancer, infections, and autoimmune diseases as they are often the initial source of T cell effector cytokines triggered prior to adaptive immune cells. Published work from the Evavold lab, and others, has demonstrated that conventional T cells become activated through TCRs that are a mechanosensor in which the affinity and bond lifetime under force determines the effector functions. Unlike the adaptive T cell response which is more nuanced, innate T cells (iNKT, gd, MAIT, or ILCs) respond to triggering antigen in a binary on/off manner. Our R21 proposal will be the first to examine the role of 2D affinity and applied force to the TCR in the cytokine response of iNKT cells and will define how these features shape development of CD8 T cell-driven ECM. The central hypothesis of our grant revolves around the idea that, like conventional ab T cells, the TCR for iNKT can be classified as a mechanosensor that integrates differences in 2D affinity and bond lifetimes under force of the presented glycolipid antigens resulting in discrete cytokine production and phenotype. Based on preliminary data, the working hypothesis of this grant is that bond lifetime under force determines effector phenotype in iNKT cells and that manipulation of iNKT cell activation in the periphery can be utilized as a novel adjunctive therapy for CD8-mediated organ-specific pathologies in malaria. We predict that conserved ligands with a lower affinity and shorter bond lifetime in P. berghei ANKA infection will favor iNKT2 cell responses and provide protection from neurological manifestations of Plasmodium infection in the ECM model. Our working hypothesis will be tested under 2 specific aims: Aim 1. Define the strength of signal exerted by the TCR of iNKT cell subsets in both naïve mice and those with P. berghei ANKA infection Aim 2. Prohibit the expansion and trafficking of pathogenic IFN-g secreting Plasmodium-reactive CD8 T cells through administration of chemotherapeutic iNKT2-inducing lipid antigens

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT / PROJECT SUMMARY Closed-mechanism peripheral nerve injuries are among the most devastating neurologic injuries, often with complete loss of functional use of a limb. Nerve regeneration, i.e., the cascade of regenerative changes after injury, commonly fails in rapid-stretch injuries. Instead, a neuroma forms – where abundant scar tissue replaces the normal pathway for nerve regeneration. Unfortunately, there is little pathophysiologic understanding of how the regenerative cascade that normally promotes nerve regrowth is corrupted into to the abundant fibrosis, chronic inflammation, limited axonal regeneration, and painful consequences of neuromas. The goals of this project are to evaluate the cellular composition, activation state and spatial niches in neuroma formation, validate our preliminary data that NETosis appears to be one of the inciting events in neuroma formation, and treat nerves with DNase I – which appears to improve nerve regeneration after injury. First, the initial response to nerve injuries appears to determine whether the nerve will recover or not. Our preliminary data highlights five expression pathways in neuroma formation: 1) Vascular 2) Neuronal 3) Fibrosis 4) Cell Death 5) Inflammation. Furthermore, we have substantial evidence that inflammation is exuberant and persistent, suggestive of a potentially coordinating role. We will analyze cell types and states, as determined by protein and genetic expression, to help identify the mechanistic details of neuroma formation. NETosis has been established, in other disorders, to lead to impaired vascular remodeling, worsening fibrosis, and amplification of inflammation - aligning with the critical features of neuromas. Our preliminary data indicates that NETs form selectively in severe, neuroma-forming injuries, and not in milder stretch injuries that do not form neuromas. Furthermore, preliminary evidence indicates that genetic reduction in NETosis (PAD4 KO) improved the gross pathology, fibrosis and axonal regeneration after severe nerve injury. We will assess the impact of reducing NETosis on nerves recovering after severe injury, using PAD4 KO animals and Cl-amidine treatment, which reduces NETosis, in comparison to untreated WT animals. Second, we will assess the impact of exacerbating NET formation in lower-severity injuries, using purified NET products or pharmacologically activated neutrophils. Third, we will assess impact of re-introducing NETs in PAD4 KO animals to assess for return of neuroma formation. Lastly, we will investigate DNase I for identification of optimized treatment strategies for severe nerve injuries. Our preliminary data suggests treatment with DNase I over 14 days dramatically improves axonal regeneration. We will investigate dosage, duration and initiation of DNase I treatment, to develop optimal dose strategies. The outcomes of the proposed experiments will (i) address a knowledge gap on mechanisms underlying neuroma formation, (ii) determine how NETs impact nerve regeneration, and (iii) provide essential data for designing future human trials with DNase I for nerve injuries.

NIH Research Projects · FY 2026 · 2025-07

PROJECT SUMMARY/ABSTRACT This K23 Patient-Oriented Mentored Award Proposal presents a structured 5-year research and career development plan to prepare the candidate, Amiko Uchida, MD, to be an independent physician-scientist and leader in the field of eosinophilic esophagitis (EoE). EoE is a chronic inflammatory esophageal disease affecting nearly 1 in 1000 US individuals and is increasing in incidence. Despite the high frequency of EoE, its pathophysiology is incompletely understood, resulting in limited treatment options and reliance on expensive endoscopies for diagnosis and monitoring. One treatment is a food elimination diet. About half of EoE patients respond to empiric removal of wheat and dairy (2FED) from the diet. While this has led to the idea that certain foods trigger an inappropriate immune response against dietary antigen(s), there is a paucity of data to support food-specific immune responses (e.g. antibodies) as the sole driver of EoE. Interestingly, eliminating more foods does not always result in more remission, suggesting that EoE may be more complex and depend on environmental factors like the microbiota. The microbiota is a crucial factor for many intestinal diseases and is well-known to be affected by diet. The effects of 2FED on the intestinal microbiota composition and metabolites, like short chain fatty acids (SCFAs), are unknown in EoE. Given our limited understanding of EoE pathophysiology, there is a critical unmet need to understand how diet induced microbial changes impact EoE to allow for novel therapies. Dr. Uchida is uniquely positioned to close this knowledge gap as she is an Adult Gastroenterologist and co-Director of the Eosinophilic GI Disease Clinic at the University of Utah conducting translational research on EoE. Dr. Uchida will build on her previous scientific and clinical training by determining how 2FED impacts the microbiome and can be leveraged in EoE. Specifically, she will 1) Characterize the fecal microbial metagenome, metatranscriptome, and metabolome associated with 2FED- induced EoE remission, 2) Define serum IL-33 and metabolite changes associated with 2FED-induced EoE remission, and 3) Delineate the contribution of SCFAs to eosinophil-intrinsic anti-inflammatory responses. Dr. Uchida will investigate these aims under the guidance of her primary mentor June Round, PhD, who is a world- renowned microbiome immunologist as well as her co-mentor Kathryn Peterson, MD MSCI, an acclaimed EoE expert and clinician investigator. Both have long histories of training successful scientists and have established a productive, integrated team with the candidate. Dr. Uchida will achieve her goals through the guidance of her mentors, a scientific advisory committee, formal coursework, seminars, and hands-on training in advanced laboratory techniques. The scientific environment of the University of Utah is particularly strong and offers numerous scientific and career development resources. Successful completion of this project will inform the development of new microbial-based therapeutics and non-invasive disease markers in EoE, and open avenues of independent research for the candidate.

NIH Research Projects · FY 2025 · 2025-07

SUMMARY Osteoarthritis (OA) is a debilitating disease characterized by loss of joint space, degeneration of cartilage at articular surfaces, remodeling of bone and other joint tissues, and inflammation. Although it is the major cause of disability in the aged population, not a single disease modifying drug is available. The functions and identities of the biological processes that pose vulnerabilities to OA are unknown. We have identified a novel association between OA and PIEZO1 activity exhibiting heightened susceptibility to age-associated OA. Our analyses of 151 families with dominantly inherited OA identified four independent OA-susceptibility alleles affecting the mechanosensitive cation channel, PIEZO1. An independent indication that PIEZO1 is critically involved in OA comes from a recent genome wide association study (GWAS) that identified a dominant rare coding allele of PIEZO1 associated with reduced OA progression. We do not know how these mutations affect PIEZO1 activity and OA susceptibility. PIEZO1 responds to mechanical cues to regulate diverse biological processes in maintaining musculoskeletal homeostasis. However, its role in OA susceptibility is unresolved. Here we combine single channel biophysical analyses with cellular and molecular studies using mice harboring gain- and loss- of function human PIEZO1 mutations to determine the role of PIEZO1 in joint homeostasis and OA. Our preliminary electrophysiological studies indicate that all four familial OA alleles are hypomorphic, reducing the open probability of the channel in response to mechanical stimulus, while the GWAS allele is hypermorphic. Mice carrying the dominant familial OA-associated Piezo11398W allele, introduced by genome editing, have significantly increased inflammatory gene expression in the knee joint. We hypothesize a reduction in Piezo1 activity is a central component of the homeostatic signaling networks and cell processes that maintain the synovial joint. In Aim 1 we identify how mutations in PIEZO1 alter single channel activity, trafficking, and cellular Ca2+ levels in chondrocytes expressing WT and mutant proteins. In Aim 2 we test the hypothesis that mice expressing Piezo11398W will have reduced susceptibly to injury induced OA while Piezo12484L will accelerate OA. Mouse models of age-dependent OA are rare and changes that indicate early stages of OA are unknown. Our preliminary data indicate that PIEZO1 has a different role in injury- vs age-associated OA. Therefore, it is crucial to define the role of PIEZO1 during aging and establish aging models of OA. In Aim 3 we establish i) that reduced Piezo11398W activity promotes OA-associated gene expression in multiple joints during aging, ii) and causes accelerated onset of histologically recognizable age-dependent OA in the joints that are correlated with use/weight-bearing activity. RNA-seq analyses will be used to uncover changes in gene expression and cell populations in the joint that tracks development of OA. Our work will have direct clinical impact, informing efforts to identify biomarkers of susceptibility or early stages of OA, which in turn help develop better therapies for OA.

NIH Research Projects · FY 2025 · 2025-07

The Medical Imaging with Deep Learning (MIDL) conference is an annual event dedicated to advancing research and innovation in medical imaging, image analysis, and the development and integration of deep learning, machine learning (ML), and AI. The scope of MIDL encompasses deep learning approaches to all medical image analysis challenges, offering a platform to explore theoretical advancements and their translational applications. By fostering interdisciplinary collaboration, MIDL addresses complex problems in healthcare and biomedical sciences. NIH funding will support registration awards for students in US based institutions to attend MIDL 2025. This initiative aims to promote access to networking, mentorship, and professional opportunities in medical imaging and AI community. Recipients will be selected based on their scientific qualifications, ensuring the awards recognize and support promising researchers.

NIH Research Projects · FY 2025 · 2025-07

Abstract The generation of long-lasting humoral immunity is the purpose of almost all vaccines. Long-lasting humoral immunity is mediated through antibodies produced by plasma cells (PCs) and memory B cells (MBCs). Despite the outstanding success of vaccinations overall, many clinical and experimental vaccines fail to induce long- lasting humoral immunity. One key example is the influenza vaccine, which requires administration every year. Upon encountering a foreign antigen, antigen-specific B cells proliferate and differentiate into the germinal center (GC) B cells and subsequently generate PCs and MBCs. B cell receptor and CD40 receptor signaling result in the activation of NFkB in GC B cells. It is well established that transcription factors NFkB cRel and RelA are critical for physiological B cell responses and that their misregulation leads to B cell-mediated diseases. cRel and RelA are essential for generating GC B cells and PCs. The activation of cRel and RelA depends on the degradation of NFkB inhibitors, IkBs. IkBa and IkBe have different binding preferences to cRel and RelA. IkBa binds to RelA-containing dimer (RelA:p50) with a higher affinity than cRel-containing dimer (cRel:p50), whereas IkBe binds to cRel:p50 with a higher affinity than RelA:p50. The network of IkBa, IkBe, cRel, and RelA is defined here as the NFkB signaling network. The role of the NFkB signaling network in regulating affinity maturation and generating PCs versus MBCs is unknown. In this project, we will define the role of the NFkB signaling network in regulating affinity maturation and in the generation of PCs versus MBCs. The objective of Aim 1 is to define how IkBa and IkBe regulate affinity maturation and the generation of PCs and MBCs. The objective of Aim 2 is to identify how cell surface receptors regulate the abundance of IkBa and IkBe and thus control cRel and RelA activation in GC B cells. The objective of Aim 3 is to determine whether the vaccine-induced response of IkBa- deficient and IkBe-deficient mice can enhance humoral immunity against influenza infection. The proposed research is innovative because it defines the discrete role of IkBa and IkBe in regulating the fate of GC B cells. This work uses newly generated cRel and RelA fluorescence reporter mice and IkBe and IkBa knock-out mice. The proposed research is expected to define the mechanism that allows the NFkB signaling network to controll different gene regulatory programs within phenotypically-identical GC B cells to regulate affinity maturation, as well as the generation of PCs and MBCs. The study will reveal new insights into the generation of long-lived humoral immunity and improve immunity against influenza.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract In this proposal, I discuss my plans to use a high-throughput yeast display platform to dissect the essential changes that occur in viral protein binding specificities. I am using the coronavirus family for my investigations because there are instances of species specificity changes of interest in the SARS-related coronaviruses that are of interest, as well as instances of wholesale receptor changes in the MERS-related coronaviruses. Thus far, this work has begun to find the substitutions of interest along the gains of human ACE2 binding in the SARS-related coronaviruses. From preliminary results, we can see one substitution that is highly associated with mutations that can bind human ACE2. In addition, I have started to parse out the important branch where the wholesale receptor change occurred along the MERS-related coronavirus family phylogenetic tree. These changes are particularly interesting because the evolutionary genetic basis for these receptor-binding specificity changes has not been characterized. Through this research, I will inform the field of the necessary changes that need to occur for viruses to switch receptors as well as what effects species specificity has on these receptor binding dynamics. Overall, this work will be important for the field, too, because it will lay a foundation of how these viruses are gaining the ability to spillover from their animal reservoirs and into humans. Zoonotic transmission, or viruses gaining the ability to bind human receptors, is a major threat to human health, as we know from the SARS-CoV-2 pandemic and the MERS epidemic, for instance.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT Natural killer (NK) cells function as cytotoxic cells towards virally-infected and tumor cells. They are also important producers of cytokines to orchestrate immune response. These functions are regulated through a balance of activating and inhibitory receptor signals in the presence of stress- and self-signals. When virally infected or malignant cells downregulate expression of MHC class I (MHC-I), inhibitory receptors on NK cells are no longer engaged, inducing NK cell function. Despite the idea that NK cells are based on such innate recognition mechanisms, NK cells demonstrate different target cellular lysis when presented with different peptides such as SIINFEKL from ovalbumin (OVA) or RGYVYGQGL from Vesicular Stomatitis Virus (VSV) on MHC-I. This differential effector function occurs in the complete absence of direct contacts between peptide and Ly49C and the basis for this is unknown. The overall goal of the proposed work is to determine how Ly49C imparts these different NK cell functions with a view to developing strategies to manipulate NK cells. Antigen-dependent NK cell recognition via modulation of inhibitory receptor recognition is a paradigm shifting concept and may impart NK cell trained immunity. The Ly49 (mouse) family of inhibitory receptors in mice have been well characterized with Ly49C being the dominant player in NK cell inhibition and licensing. Ly49C binds to MHC-I H-2Kb to signal self and prevent the breakdown in tolerance. Ly49C binds H-2Kb underneath the peptide-binding domain alongside the α3 helix domain with no direct contacts to the H-2Kb loaded peptide. As an innate germline encoded receptor, non-specific binding to the constant region of MHC-I would be consistent with the nature of innate immunity. However, Ly49C-peptide-H-2Kb interaction results in different biological activity depending on the peptide. This project is based on published and preliminary data showing: 1) no peptide-dependent structural differences with peptides bound to H-2Kb or when the entire Ly49C-peptide-H-2Kb interacts; 2) mechanosensing measures such as two dimensional bond lifetime under force can dictate differential cell responses and, 3) our preliminary data demonstrating that mechanosensing properties of inhibitory receptor Ly49C and its interaction with peptide-H-2Kb is different depending on the peptide. My working hypothesis is that the bond lifetime as it occurs at the membrane allows Ly49C to distinguish different peptides loaded into MHC. I will test my working hypothesis under the following 2 specific aims: Aim 1. Demonstrate that Ly49C bond lifetime with peptide:MHC molecules is a key feature for NK cell function. Aim 2. To delineate critical amino acids of peptide for Ly49C function. At the completion of the proposed work I expect to define how NK cell trained immunity occurs in an antigen specific manner. This information could be used for rationally designed engineering for the treatment of cancers that can be destroyed by NK cells or infectious diseases where peptide priming of NK cells could lead to trained NK cells that could better fight infectious diseases.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY This MIRA/R35 application is designed to replace our NIGMS R01 GM147310-01 and a recent NIGMS R01 application that was scored on first submission but rejected based on not being able to have two simultaneous NIGMS R01s. Following NIGMS advice, I have consolidated these projects into this MIRA application. Our overarching goal is to uncover fundamental cell signaling mechanisms that can be therapeutically targeted in disease. All projects began with PTM discovery approaches and have now grown into multi-pronged efforts that span molecular biology/biochemistry, structural biology, and drug development. The first project addresses the regulation and function of kinases in the understudied ‘dark kinome’, with current focus on an unusual ubiquitin- binding non-receptor tyrosine kinase called TNK1 (currently funded by GM147310-01). C-terminal truncations in TNK1 convert it into an oncogenic driver, yet it’s normal “day job” function and mechanism of regulation are still poorly understood. We also developed the first TNK1 small molecule inhibitor, TP5801, which has passed IND hurdles for phase-I trials but lacks clear clinical direction due to our still nascent understanding of TNK1 biology. Thus, our research addresses critical gaps in TNK1 function and regulation that will inform the clinical path of TP5801. Our work on TNK1 takes advantage of cell-based and in vitro biochemical systems recently developed in the lab, and also extends to mouse models in which we have CRISPR-engineered constitutively active (by disrupting 14-3-3 binding) or kinase-dead tnk1 alleles. Together, our preliminary data point to a role for TNK1 in sensing poly-ubiquitin and instigating inflammatory signaling, and also help explain how genomic rearrangements aberrantly activate the kinase. The second project addresses key gaps in our understanding aggrephagy, a form of autophagy that occurs in nutrient replete conditions and rids cells of toxic protein aggregates that could otherwise cause degenerative proteopathies (e.g., ALS). Specifically, we focus on the earliest steps—how the lipid scramblase ATG9A is recruited to sites of aggrephagy, referred to as “ubiquitin-rich condensates”, and engages with autophagy/aggrephagy machinery to encapsulate and degrade protein aggregates. This project also addresses how ubiquitin-rich condensates can act as platforms to assemble pro- inflammatory (and other) signaling complexes, providing a link between aggrephagy and inflammatory signaling that could explain why persistent, pathological inflammation is a hallmark of proteopathy diseases. A third related project extends our work on aggrephagy to understand how disruption of ATG9A-mediated aggrephagy sensitizes cells to nucleic-acid sensing pathways that activate interferon (IFN) signaling. Together these aggrephagy-focused projects uncover fundamental mechanisms and links to inflammation that could be exploited for new approaches to manipulate IFN signaling in disease.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Mutations during embryonic development, aging, cellular metabolism, and environmental exposure are permanently recorded in the genomes of each cell and its daughters. Depending upon whether the mutations can be detected in regular next-generation sequencing, they are recognized as clonal or non-clonal in nature and present different features. Elucidating the patterns of these mutations and their potential to transmit to offspring is key to understanding congenital de novo mutation (DNM) disorders and genetic variability across human generations. As parents age, the number of DNMs in their germ cells increases, and with this, an increased risk of DNMs and the disease they cause in offspring. Although age-related DNM risks have been reported in large populations, our understanding of how paternal-specific clonal and non-clonal mosaicism contribute to offspring and how natural selection shapes the mutation pattern is still limited. During my previous graduate and postdoctoral research, I established the concept that a considerable portion of DNMs in children with neurological and psychiatric disorders arise from clonal mosaic mutations in the sperm (Yang, et al. Cell 2021). I developed experimental and computational pipelines to accurately detect clonal mosaic mutations in bulk samples with deep whole-genome sequencing (Yang, et al. Nature Biotechnology, in press; Breuss, Yang, co-firsts, et al. Nature 2022). In this K99/R00 application, I aim to unravel the feature of the non-clonal gonadal mutation burden at the single-cell level, employing multidisciplinary approaches spanning the mentored [K99] and independent [R00] award phases. I will compare the genomic sequences from 700 single human sperm from bulk sperm sequences in 35 healthy young men, analyze the genomic positions where the non-clonal mutations tend to reside compared to the clonal ones, and study the impact of those mutations (Aim 1). I will develop new computational software to accurately detect mosaic mutations from single cells not only from haploid and diploid genomes, and develop experimental approaches to accurately validate the somatic mutations from single-cell amplified DNA (Aim 2). Finally, I will measure the single-cell DNA mutation rate and mutation patterns using 2300 single sperm from an additional 45 young versus 75 aged donors for clues on age-related mutational mechanisms and how they will impact the genome stability in the next generation before and after natural selection (Aim 3). Overall, the results from this proposal will help us to understand the non-clonal mosaic mutational burden, mutation distributions, as well as mutational functions in human sperm, and the age-related genetic impacts on the genome stability of the next generation. My career goal is to lead an independent research group focusing on somatic mutations in the human genome, their causes, and predicting their consequences on child health. During the K99 phase, I will continue to receive subject recruitment, reproductive science, experimental, computational, and career development training from my postdoctoral advisor Dr. Gleeson, co- mentors Dr. Wilkinson and Sebat, as well as external mentors at UC San Diego and other institutes. The rigorous mentored support will greatly my knowledge in human subject handling and reproductive science, as well as getting me prepared for job applications. The results obtained in the K99 phase will facilitate my transition to an independent investigator in the R00 phase and lay the foundation for my future career.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY / ABSTRACT Persistent balance problems are common sequelae after mild traumatic brain injury [mTBI]. Current approaches to treat these balance problems traditionally focus on principles of sensory integration derived from vestibular rehabilitation. However, recent evidence using destabilizing perturbations suggests that balance deficits may originate from motor impairments, rather than sensory impairments. This reactive balance – the ability to recover stability after an external perturbation – is an important domain of balance that is distinct from static (i.e., standing) and dynamic (i.e., walking) balance tasks. Preliminary studies suggest this reactive balance is impaired after mTBI, but the components underlying these impairments and the extent to which these impairments manifest during daily life remain unclear. While reactive balance can be improved through perturbation-based balance training in other populations, current clinical practice guidelines for the treatment and management of mTBI provide no guidance on treating reactive balance. The objective of this study is to understand the components underlying reactive balance deficits in people with mTBI and their impact on daily life. The long-term goal is to guide targeted rehabilitation approaches that incorporate rapid motor responses and reactive balance training to improve the function and quality of life for people with imbalance after mTBI. Using a sliding platform and mechanized shoes, we will examine how mTBI affects reactive balance during standing and locomotion, including walking and turning. Specifically, we will examine the two important components of reactive balance: the ability to prepare (i.e., prime) a motor response in anticipation of an upcoming perturbation and the ability to execute a motor response after a loss of balance. We will also develop a method to assess reactive balance during daily life using wearable inertial sensors to assess reactive balance during free-living daily mobility. Our aims will provide component-specific (Aim 1) and task-specific (Aim 2) targets for future rehabilitation while exploring reactive balance during daily living (Aim 3). Combined, these aims will provide the foundation for new, perturbation-based balance rehabilitation after mTBI, including innovative methods to assess reactive balance during daily living, that can complement current standard-of- care.

NIH Research Projects · FY 2025 · 2025-07

There are an estimated 6-7 million people with opioid use disorder (OUD) in the United States (US). Medications for OUD is the gold standard treatment, but the full benefits of these medications remain unrealized, due to several factors, including inadequate access and poor retention. Extended-release buprenorphine (XR) injection for OUD became available in the US in 2018, offering additional choice and flexibility. XR has the potential to improve access and retention for some patients because of advantageous characteristics (e.g., no burden of daily dosing, no risk of diversion or misuse, less fluctuation in daily drug levels). However, compared to daily sublingual buprenorphine (SL), XR costs more and has more restrictive insurance coverage, which may exacerbate persistent patterns of medication access and retention. There is currently little guidance informing treatment decisions between XR and SL buprenorphine. A better understanding of which patient subgroups benefit most from XR is critical to inform optimal treatment for each person and motivate efforts to address barriers to access. The goal of this K99/R00 is to address these gaps and to support Dr. Ross’s career goal of becoming an independent investigator whose research advances evidence-based treatment of OUD through cross-disciplinary collaboration and application of novel quantitative methods. Through the award, Dr. Ross will build on her training as an epidemiologist with experience in pharmacoepidemiology and causal inference to develop competency in 1) OUD and its treatment, 2) conceptualization and quantification of population subgroup differences in health services, 3) statistical methods for precision medicine, and 4) professional skills to be a responsible and independent investigator. The proposed research will use recent Medicaid data from 50 states and data from the large multi-site Optimizing Retention, Duration, and Discontinuation Strategies for OUD Pharmacotherapy trial (RDD). The research aims are to: 1) assess current demographic differences in real-world XR treatment for OUD among Medicaid patients, 2) identify the patient subgroups expected to have the greatest improvement in treatment retention from XR relative to SL using the RDD trial data, and 3) predict the effect of personally tailoring buprenorphine formulation decisions on real-world treatment retention and demographic differences in retention among Medicaid patients. This work is aligned with NIDA’s strategic priorities to support personalized addiction medicine and leverage data science tools for clinical decision making. This grant will support Dr. Ross’s transition to an independent investigator in OUD treatment through salary support, focused training, and a dedicated mentorship team.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY / ABSTRACT Our long-term goal is to improve the health, care, and well-being of individuals with transhumeral limb loss by uncovering the biomechanical origins of functional deficits and long-term musculoskeletal complications that disproportionately affect them. Trauma is the most common cause of transhumeral limb loss, and while socket- based prosthetics are standard of care, more than half of affected individuals wear their devices infrequently or abandon them altogether. This negatively affects quality of life and results in overuse injuries and pathologies in the intact contralateral limb. Prior research has examined prosthetic devices/interfaces, outcomes of treatment, and percutaneous osseointegrated devices (PODs) to skeletally attach prostheses since they may mitigate socket issues and restore normal shoulder function. Yet there are virtually no data on three-dimensional (3D) shoulder kinematics and morphology after unilateral transhumeral amputation. Given the paucity of data on these understudied and underserved individuals, we will collect high precision, bilateral, 3D form-function data to quantify local (e.g., bone) and global (e.g., joint level) 1) kinematic compensations and 2) morphologic variations in the shoulders of patients who use socket suspension. We will then 3) analyze the form-function relationships that arise from these data. This cross-sectional observational study will recruit unilateral transhumeral amputees and a healthy control cohort. A medical history and medical imaging of both shoulders will be collected. Dynamic 3D motion of the thorax, scapulothoracic and glenohumeral joints in the shoulders, and the arm/prosthesis, will be captured using optical motion analysis and dynamic stereoradiography. Anatomy will be quantified on medical imaging using established 2D/3D clinical measures, and 3D anatomic models will be used to create statistical shape models that uncover previously unseen modes of morphologic variation. Analyses will examine bilateral (a)symmetry within and between groups to quantify biomechanics related to functional deficit and musculoskeletal complications, controlling for age, sex, residual limb length, and time since amputation. The results of these studies will provide clinicians, device designers, and researchers with the first accurate subject- specific in vivo 3D form-function measurements in transhumeral amputees. These extremely rare, yet highly valuable data will facilitate discovery of morphologic and kinematic biomarkers related to pathology and functional deficiency in shoulders after transhumeral amputation. Research into prosthetics and implanted device design, bone-limb-prosthesis interfaces, musculoskeletal function and rehabilitation, and surgical planning will all benefit. Techniques like finite element analyses and musculoskeletal modelling can then also query joint contact patterns and load/deformation of tissues using subject-specific data to drive studies. These data will also motivate and benchmark FDA evaluation of PODs for safe and effective widespread clinical use for limb loss.

NIH Research Projects · FY 2025 · 2025-07

This project addresses the lack of access to evidence-based childhood obesity interventions (EBI-CO) in rural areas where both poverty and obesity is prevalent. Building Healthy Families (BHF) is an adapted EBI-CO designed for, and implemented in, rural areas and has successfully achieved clinically and statistically significant reductions in child BMI z-score (-0.27±0.22)—a similar magnitude of effect relative to previous efficacy trials. To increase the likelihood of broad dissemination and implementation of BHF to other rural communities we piloted the development and implementation of the BHF Online Training Resources and Program Package (BHF Program Resources). The BHF Program Resources is a ‘turn-key’ online resource that includes a train-the-trainer system, program materials and a data portal for use by community-based implementation teams. In our pilot study we compared the use of the BHF Program Resources with and without community participation in an action learning collaborative (LC) in changing RE-AIM outcomes. We found the learning collaborative was related to improved Reach (94% vs 74% attendance), Effectiveness (BMI z-score change -0.15±.08 vs -0.09±0.11), and Implementation (91% vs 73% adherence to protocol). Further, Adoption and Maintenance were successfully achieved, but did not differ by condition (100% of enrolled communities initiated the program and planned a second cohort of families). We propose to expand our pilot into a fully powered Hybrid Type 3 effectiveness-implementation, cluster RCT to test the utility of the BHF Program Resources Only (BHF-PO) with and without a BHF Learning Collaborative (BHF-LC). We will scale up a bundled adoption strategy we used successfully in our pilot to engage 30 micropolitan and rural communities across the 6-state region served by the Mountain West Family Healthy Weight Collaborative and Huntsman Cancer Institute (NM, ID, MT, NV, WY, UT; See Letters of Support). We will randomly assign communities to either the BHF-PO (n=15) or in combination with the BHF-LC (n=15) to determine the relative effects on RE- AIM outcomes of reach, effectiveness (secondary outcome), implementation (primary outcome), and program maintenance. In addition, we will assess potential mechanisms of change across RE-AIM outcomes using the Integrated Promoting Action on Research in Health Sciences (iPARIHS) framework. Finally, economic evaluation will be conducted to determine costs and benefits across RE-AIM outcomes.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Regulatory T cells (Tregs) are essential for maintaining immunotolerance, protecting tissues from immune- mediated damage while preserving normal immune function. Chimeric Antigen Receptor T regulatory cells (CAR Tregs) have emerged as a cutting-edge therapeutic approach due to their enhanced specificity, potency, and reliable sourcing. However, despite their promise, current CAR Tregs have struggled to achieve durable tolerance in clinical applications, exposing critical gaps in efficacy and highlighting the need for innovative strategies to improve their therapeutic potential. This project introduces a transformative "3-In-One" CAR Treg design to address these limitations. Unlike conventional CAR Tregs, which rely on disease-specific antigens for activation and suppression, the "3-In-One" approach utilizes effector T cells (eTconv) as a multi-functional target. In this design, eTconv serve as the attractant, activator, and suppression focus of CAR Tregs, thereby streamlining their activation and enhancing their suppressive efficacy. This strategy was crafted based on core working mechanisms of CAR Tregs that center around eTconv rather than antigens and is expected to significantly improve the efficiency and durability of CAR Treg-mediated immunotolerance. We propose using Programmed Death-1 (PD-1), a marker predominantly expressed on activated eTconv, as the target for this innovative CAR Treg design. PD-1CAR Tregs are expected to exhibit superior suppressive efficacy compared to traditional antigen-specific CAR Tregs by selectively suppressing eTconv without affecting naïve T cells or normal immune responses. This targeted approach offers a significant advantage over conventional CAR Tregs, whose application is often restricted to specific antigen-driven diseases. In this R21 project, we will develop and evaluate PD-1CAR Tregs with varied affinities for PD-1, alongside a reference MOGCAR Treg that targets a specific myelin antigen. Our research aims to: I. Characterize PD-1CAR Tregs. We will assess binding specificity, regulatory phenotypes, cytokine secretion, suppression of eTconv, and bystander immunosuppressive effects. II. Evaluate Therapeutic Efficacy and Safety. Using two experimental autoimmune encephalomyelitis (EAE) models representing distinct autoimmune conditions, we will measure the ability of PD-1CAR Tregs to restore immunotolerance. Outcomes will include clinical EAE scores, suppression of demyelinating eTconv activity. A critical advantage of PD-1CAR Tregs is their potential applicability across multiple antigen-driven diseases, avoiding the requirement to develop different CAR Tregs for different medical conditions. Furthermore, we will rigorously evaluate whether the immunotolerance induced by PD-1CAR Tregs selectively targets pathogenic responses without impairing host immunity to infections. In summary, the "3-In- One" design represents a paradigm shift in CAR Treg therapy, combining efficiency, specificity, and versatility. By targeting eTconv via PD-1, this approach promises to revolutionize CAR Treg-based treatments for autoimmune diseases and other conditions requiring immune tolerance.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY We propose the Huntsman Cancer Institute (HCI) Oncological Data Science (ODSi) Training Program. HCI is a National Cancer Institute-designated Comprehensive Cancer Center and the official Cancer Center of the State of Utah. Contributing to HCI’s mission of reducing morbidity and mortality from cancer, ODSi provides a robust two-year predoctoral training program that leverages HCI research and infrastructure strengths in bioinformatics, molecular population health, and, jointly cancer data science, plus integrates trainee interactions with cancer clinicians and our Disease Centers, to provide broad multidisciplinary training in cancer biology. The program goal is to prepare young scientists to become data ambassadors and data innovators in the use of biomolecular-anchored data science techniques and methodologies to advance the understanding, prevention, diagnosis, and treatment of cancer. We have carefully selected 27 experienced HCI members to serve as training faculty, with an additional 8 promising Assistant Professor mentors. The ODSi program will support six predoctoral candidates per year. Trainees will be recruited into mentor’s laboratories after entering the University of Utah (U of U) via graduate programs in Molecular Biology, Biomedical Informatics, Public Health, Population Health Science, and our MD-PhD program. These graduate programs recruit an average of 57 students annually, ensuring a deep pool of potential ODSi participants. ODSi trainees will be selected based on their academic performance, promise as cancer researchers, and commitment to cancer data science training. Training in the ODSi program includes courses in cancer data science, basic and clinical cancer biology, bioinformatics, epidemiology, and data science electives. To complement these didactic courses, our trainees will have multiple opportunities to understand cancer from a clinical and patient-centric perspective. These opportunities provide broad exposure to cancer as a disease, including exposure to different operational units within HCI, physician shadowing, and attendance at Molecular Tumor Boards. These activities are buttressed by Research-In-Progress presentations, a career development series, and participation in an annual ODSi trainee retreat. The ODSi program is strengthened by optional training activities, including internships at local biotechnology companies and a Master of Science in Clinical Investigation. ODSi trainees will receive experimental and career development advice from their mentors and their thesis committee, guided by required Individual Development Plans. The ODSi leadership team will receive guidance from an External Advisory Board comprised of leaders of successful NCI T32 programs and other cancer data science training, and an Internal Advisory Board comprised of U of U education leaders. We will use professionally-developed evaluation tools to ensure the effectiveness of the ODSi program, and we will track trainees for ten years following completion of the program.

NIH Research Projects · FY 2026 · 2025-06

Abstract: This is a T32 submission by the University of Utah Division of Clinical Pharmacology to the NICHD-sponsored Clinical Pharmacology Training Network (CPTN). The goal of the CPTN training programs is to ensure that a highly trained workforce is available in clinical pharmacology and therapeutics. Pediatric and maternal pharmacologists are urgently needed because the majority of drugs prescribed to children and pregnant and lactating women lack dosing information specific to these populations. The lack of evidence-based dosing results in off-label prescribing and places these populations at risk for treatment failures and toxicities. These outcomes can be avoided with innovative clinical drug studies. Design and interpretation of these studies require trained pediatric and maternal pharmacologists. Properly trained pediatric and maternal clinical pharmacologists are indispensable for their ability to integrate basic and clinical science including trial design, epidemiology, biostatistics, biomarker qualification and standardization; disease-specific biomarker development; novel clinical trial design and endpoints; and bioinformatics. At present there is a shortage of scientists who have formal training in pediatric and maternal clinical pharmacology. The goal of the Utah Pediatric and Maternal Clinical Pharmacology T32 Fellowship Program is to develop clinician- and PhD-scientists who will be leaders in the field of pediatric and maternal clinical pharmacology research. This will be accomplished by providing training and experience in the methods and conduct of basic and clinical drug research in the different phases of pediatric drug development and fostering interaction with pediatric subspecialists to address deep knowledge gaps in pediatric therapeutics. We will leverage the resources of the Division of Clinical Pharmacology in the Department of Pediatrics (expertise in pediatric clinical pharmacology), the Department of Obstetrics and Gynecology (expertise in maternal pharmacology), the Department of Population Health Sciences (expertise in pharmacoepidemiology), and the College of Pharmacy (expertise in assay development and the mechanistic methods that inform clinical pharmacology studies). We have an excellent team of 22 established and emerging mentors who will support up to 3 postdoctoral positions per year. The T32 fellowship leverages the infrastructure of the existing American Board of Clinical Pharmacology-accredited Utah Clinical Pharmacology Fellowship Program. The T32 fellowship will combine expert mentorship with rigorous training in the principles of clinical pharmacology, quantitative skills, clinical trials, regulatory affairs, and writing and presentation. Training will include a mix of didactic coursework and practical training. Each fellow will have their own primary project and be integrated into multiple other projects. At the end of the two-year training, each fellow will have answered an important clinical question in pediatric or maternal pharmacology, have 5-10 publications, have submitted one grant, and will be ready to submit a mentored K-level grant.

NIH Research Projects · FY 2026 · 2025-06

––– PROJECT SUMMARY/ABSTRACT –––––––––––––––––––– R01: Psychostimulant responses and sleep. –––– Substance use disorder (SUD) and sleep alterations are reciprocally connected in a vicious feed forward cycle. In one direction, repeated exposure to addictive substances affects sleep timing, duration, and quality. The effects of drugs on sleep can even last into the abstinence period, during which patients experience irregular sleep and insomnia, and this persistent sleep disturbance is a strong risk factor for relapse to drug use. Conversely, sleep disturbances cause stress and mood imbalance and are a risk factor for the development of SUD. This includes sleep disturbances caused by means other than drugs of abuse, such as shiftwork. Some neurobiological systems are known to influence sleep as well as motivation and reward. These systems are often targeted by drugs of abuse, and a substantial body of literature has focused on the disruptive effects of acute exposure to substance of abuse on sleep. However, mechanistic studies of long-term sleep irregularities after drug exposure, as well as of how sleep disturbances affect SUD-related behaviors remain scarce. This application responds to the one-time RFA-DA-25-045 “Mechanistic studies to investigate the interrelationship between sleep and/or circadian rhythms and substance use disorders”. The goal of this proposal is to identify genes and molecular mechanisms that affect psychostimulant-induced sleep irregularities as well as sleep deprivation-caused changes in experience-dependent amphetamine preference (EDAP). We will do so using the vinegar fly, Drosophila melanogaster, an established genetic model organism with considerable economy of scale and excellent experimental amenability. Based on preliminary data showing that recovery from acute amphetamine exposure, which suppresses sleep, leads to sleep rebound, we propose to 1) determine mechanisms of stimulant-induced sleep changes, including anatomical as well as genetic investigations. 2) We will use within-fly amphetamine exposure to screen Drosophila orthologs of 2 094 human brain-enriched transcripts for their role in psychostimulant-induced sleep changes. Lastly, we have recently developed a self- administration assay that monitors EDAP with high temporal resolution, where flies develop preference for amphetamine within ~12 h, starting from indifference or slight aversion. EDAP depends on Drosophila’s canonical dopaminergic ‘reward system’. Our preliminary data show that mechanical sleep deprivation accelerates the development of EDAP and promotes EDAP at lower, normally sub-threshold amphetamine concentrations, as observed in mammals. We will 3) determine circuit and genetic mechanisms of sleep deprivation-induced changes in amphetamine preference. Results from these investigations will offer RFA- relevant “insights into the fundamental processes that link SUDs to disorders of sleep…and vice-versa”.

NIH Research Projects · FY 2026 · 2025-06

PROJECT ABSTRACT The opioid crisis remains an ongoing public health concern as annual deaths due to opioid overdose remain at record-high levels. Chronic pain contributes to both non-fatal and fatal opioid overdoses, and the lack of effective and safe non-opioid options for long-term pain management is a significant barrier to the treatment of substance use disorders. There is an urgent need to better understand the factors linking co-occurring pain and substance use to poor functional outcomes, such as overdose and pain-related suicidality. Both the sensory and affective components of chronic pain are essential to consider as both contribute to the process of pain embodiment. Pain embodiment is a part of the chronic pain experience that is characterized by a tendency to apprise bodily sensations as a threat to bodily integrity. Chronic pain patients with high pain embodiment may experience more pain-related anxiety and may be more motivated to use opioids as a safety-seeking behavior to escape pain- related distress. Here, we propose to characterize pain embodiment and how it relates to risk for opioid misuse and brain network changes as measured by resting-state functional magnetic resonance imaging (fMRI). We will study healthy individuals, individuals with chronic pain, and individuals recovered from chronic pain with a state- of-the-art imaging protocol and a comprehensive set of pain and clinical measures. Our central hypothesis is that, in chronic pain, pain embodiment increases the risk for opioid misuse and that this relationship is reflected in networks shared by addiction and central pain processing: the default mode network (DMN) and the salience network (SN). Aim 1 will identify concepts of pain embodiment that relate to opioid misuse risk. Aim 2 will determine how pain embodiment and opioid misuse risk are reflected in functional connectivity changes in the DMN and SN. Aim 3 will determine how pain embodiment and opioid misuse risk influence concentrations of proton brain metabolites in the anterior cingulate cortex, a major hub of the SN. Together, this proposal will illuminate how pain embodiment is reflected by changes in central pain processing and offer novel insights into the mechanisms contributing to poor functional outcomes in co-occurring chronic pain and opioid misuse. An interdisciplinary team of mentors will provide training in clinical and psychological behavioral measures regarding chronic pain as well as training in magnetic resonance imaging methods, including acquisition and data analysis. This training will be complemented by clinical mentorship in psychology and physical medicine and rehabilitation. This training plan was developed in collaboration with my Sponsor and Co-Sponsor to complete the central goals of conducting independent, collaborative research; advancing communication and grant-writing skills; effectively mentoring and teaching trainees in a supportive environment; and honing clinical skills in preparation for the return to medical school. This training is ideal for a future physician-scientist with the goal of improving patient care at the intersection of translational research and the clinical management of co- occurring substance use disorders and chronic pain conditions.

NIH Research Projects · FY 2026 · 2025-05

Ross River (RRV) and Chikungunya (CHIKV) viruses are emerging pathogens that target the musculoskeletal system and a major health concern as they cause explosive outbreaks in humans. RRV and CHIKV are mosquito-borne enveloped RNA viruses that cause severe arthralgia, arthritis, myalgia and myopathy that typically resolves within weeks of infection, although a significant fraction of RRV and CHIKV patients develop debilitating chronic disease. While the etiology of virally-induced arthralgia and arthritis has been intensely studied, the mechanisms underlying myalgia and myopathy are poorly understood. Our collaborative research uniquely combines the expertise of a skeletal muscle biology lab (Gabrielle Kardon) and a viral pathogenesis lab (Deborah Lenschow) to determine how RRV and CHIKV affect muscle structure and function to cause myopathy and test if and how infection of myogenic cells drives acute and chronic disease. Using new mouse models of RRV and CHIKV disease, recombinant versions of RRV and CHIKV to track infected cells, and mouse genetic alleles to label and manipulate myogenic cells we will test two hypotheses. First, we will test whether infection and cytopathic destruction of myofibers and a protracted regenerative response are the cause of acute myopathy. Second, we will test whether mechanically compromised myofibers and functionally and transcriptionally altered muscle stem stems are major causes of virally-induced chronic myopathy. Our research will elucidate the mechanisms causing the acute and chronic myopathy that are key aspects of increasingly common and debilitating RRV and CHIKV disease. In addition, our studies, which include comparison to regeneration following hypercontraction (BaCl2) injury, will provide important insights into the mechanistic differences between skeletal muscle recovery after pathogenic and acute sterile injury.

NIH Research Projects · FY 2025 · 2025-05

Abstract Funds are requested toward the purchase of a JEOL CRYO ARM 200 microscope with Direct Electron Apollo detector. The University of Utah and regional partners Brigham Young University and Utah State University have a large, vibrant, and well-funded community of structural biologists with well-established and emerging research programs that depend upon cryo-electron microscopy (cryo-EM). This community includes 13 Major Users and 11 Minor Users, supported by 35 NIH awards, who would use the new instrument. Multiple additional user grants will also directly depend upon access to the new instrument, including from NSF and private foundations. Our community already makes extensive use of the University of Utah Core Facility in Electron Microscopy, which is supported by a faculty-level Director and a faculty-level Director of Cryo-EM, both of whom have extensive experience in the instrumentation, methods, and applications of cryo-EM. The EM core facility currently houses just one instrument that is suitable for single-particle high-resolution cryo-EM, a ThermoFisher Scientific Titan Krios with a Gatan K3 detector. This instrument is state-of-the-art but is fully utilized and is experiencing a dramatic reduction in availability for single particle cryo-EM owing to commitments to the agency (Beckman Foundation) that funded purchase of both the Krios and a recently installed ThermoFisher Aquilos 2 Cryo-FIB. Specifically, the Beckman funding required that 50% of the Krios time be available to support cryo-electron tomography (cryo-ET) of samples generated using the Aquilos. This is appropriate because the Krios is especially well suited for cryo-ET. Compounding the need for enhanced single particle cryo-EM data collection, our aging ThermoFisher Tecai F20 instrument, which in principle could be used for screening, has become outdated, problematic to maintain, and will be retired. Furthermore, our user community is burgeoning with multiple recent junior recruits, recently recruited and highly active senior faculty members, and with multiple additional faculty recruitments envisioned. To address these needs, we propose to purchase a JEOL CRYO ARM 200 microscope with Direct Electron Apollo detector that will be housed in the EM core facility. This is an optimal solution because the instrument combines highly efficient screening with high-resolution data collection, is highly cost effective, can be used essentially 24/7, and will be housed, maintained, and managed in an outstanding existing core facility.

NIH Research Projects · FY 2026 · 2025-05

Project summary Understanding the genetic basis and molecular mechanisms of evolutionary conflicts, and how they shape the evolution of genomes, cells, and species is a fundamental goal in biology. In particular, our research program focuses on understanding how molecular arms races involving selfish genetic elements such as segregation distorters and transposable elements can lead to the rapid evolution of essential cellular machinery and contribute to the origins of new species. Using a highly multidisciplinary approach, we aim to understand the genetic and molecular bases of conflict and hybrid incompatibilities in Drosophila. This proposal briefly describes three research directions pursued in my laboratory. First, we aim to understand the molecular mechanisms of the lethality of hybrid males in crosses between Drosophila melanogaster and sister species. Here, I highlight our discovery of the role of rapidly evolving DNA damage-induced cell cycle checkpoints and DNA replication licensing machinery in hybrid inviability. We propose a testable cell biological model where hybrid lethality is caused by cells attempting to divide before their genomes are fully replicated. Second, we aim to understand the molecular mechanisms of meiotic drive and hybrid sterility in very young subspecies of Drosophila pseudoobscura. Here, I highlight our discovery of the existence of male germline checkpoints and a rapidly evolving non-coding RNA as the basis of drive and sterility. Third, we aim to understand the genetic and molecular basis of meiotic drive in the selfish Sex-Ratio chromosome in D. pseudoobscura. Here, I highlight our discovery of the involvement of rapidly evolving meiotic cohesin and dynein as the genetic basis of meiotic drive. We propose that a failure to properly pair and segregate the Y- chromosome during meiosis is the molecular mechanism of drive. Each of these research directions involves the development of innovative genetic mapping methods and systematically dissecting phenotypes generated by evolution down to their cellular roots; each of these directions has led us to original discoveries involving fundamental cellular processes. Through the unification of the fields of evolutionary genomics and cell & developmental biology, our research program is providing exciting advances in solving long-standing and fundamental problems in biology.