University Of Minnesota

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract Sepsis is a complex, heterogeneous, global inflammatory response to an infection that precipitates critical hypotension, is beset with high mortality, and constitutes a clinical emergency. Large blood vessels (macrovessels) become atonic and fluid-permeable, while small vessels (microvascular beds) do not perfuse, causing shock and hypoxic organ damage, a condition termed vasoplegia. Dr. Wise endeavors to become an independently funded, collaborative surgeon-scientist by characterizing physiologic injury and defining cellular processes that occur during the vasoplegia precipitated by sepsis. While elimination of the infectious pathogen is most critical to survival, mitigating vasoplegic physiology prevents morbidity to vital end organs such as the lung. Examining the problem of vasoplegia from the lens of the vascular system necessarily requires career development in three key areas, identified to optimally address the research question: cell biology and multi- omics, use of intravital surveillance techniques (microscopy), and facility with large animal models to best recapitulate human physiology in sepsis. This project is well-positioned to succeed due to a complementary mentorship and advisory team led by Dr. Greg Beilman (University of Minnesota), strong institutional support, and a focused career development plan inclusive of germane coursework, didactics, seminars and meetings. Vasoplegia will be studied using a newly-piloted porcine “fecal clot” (with ischemia-reperfusion injury) model of surgically-induced sepsis. In the first Aim, fecal clot sepsis or “sham” sepsis will be induced, followed by surgical harvest of mesenteric and saphenous arteries and veins after 24 hours. Cut rings from these vessels will be suspended in a muscle bath to determine relative severity of injury of smooth muscle cells, endothelial cells and extracellular matrix. Permeability will be assessed via immunohistochemical staining for glycocalyx. Regulation of the endoplasmic reticulum stress response and P38 MAP Kinase activation by putative endogenous inhibitor Niban will be studied, to assess the role of “off-signaling” in propagating septic inflammation. In the second Aim, lung microvasculature will be imaged using intravital orthogonal polarized spectral video-microscopy, to quantify the loss of perfused vessels and blood flow induced by fecal clot sepsis. After synthesis of macrovascular and microvascular injury patterns of vasoplegia from the first two Aims, a pilot mitigation strategy will be tested in the third Aim. Intravenous “NiPp,” a rationally designed cell-permeant phospho-mimetic of Niban developed in the laboratory of Colleen Brophy (co-mentor), will be administered to septic pigs, to decrease harmful “feed-forward” activation of injury-induced pro-inflammatory pathways leading to vasoplegia. The primary outcome will be improvement in a validated clinical porcine sepsis score. All pigs will have systemic vascular tissue extensively bio-banked upon euthanasia. The robust animal model, data to be collected, and the learned research techniques will collectively predicate a dynamic research program, with potential for translating our findings to ultimately improve survival from sepsis, a major unmet clinical need.

NIH Research Projects · FY 2025 · 2024-07

The mission of the Clinical and Translational Science Institute (CTSI) predoctoral T32 Program is to contribute to the nation’s biomedical workforce by training a diverse pool of scientists in clinical and translational science (CTS) and leadership. We will meet this mission by developing, implementing, and continually improving a training program for predoctoral trainees (Scholars) that integrates a mentored research experience with an individualized curriculum combined with professional development activities highlighting our institutional strengths in community engagement, data science, team science, and effective communication skills. The UMN CTSI T32 Program is built upon our current successful Translational Research and Career Development (TRACT) TL1 program, which guided the development of early-stage CTS researchers over the past 4 years. Through personalized mentorship, coursework, seminars, and other tools, the TRACT program has guided 18 predoctoral Scholars along a rigorous path of professional development in CTS. Moving forward, the new UMN CTSI T32 Program’s mission is to educate and train a diverse group of Scholars from across the UMN system in the fundamentals of CTS through a 2-year comprehensive yet individualized program that complements their education and training in their interdisciplinary doctoral programs. We will recruit and train 3 cohorts (diverse in discipline and underrepresented populations) of 8 predoctoral Scholars from across UMN. The UMN CTSI T32 will provide a research experience with mentors trained in evidence-based mentoring and collaboration with community members throughout the research process, inclusive of a Community Mentor and team science across the UMN ecosystem. The UMN CTSI T32 will also include near-peer learning experiences with other CTSI program Scholars, as well as provide education and opportunities for a variety of career paths. The UMN CTSI T32 will use a comprehensive and adaptive evaluation plan for rigorous programmatic assessment and continuous quality improvement to maximize the training experience. The principal Scholar outcome is to have a deep knowledge in the science of translation beyond translational research. Another key Scholar outcome is an understanding of the importance of, and key strategies for, building truly collaborative interdisciplinary teams composed of people from diverse backgrounds and experiences whose collective roles synergistically optimize the advancement of diagnostics, therapeutics, clinical interventions, and behavioral modifications that improve health. Other outcome measures include the ability to communicate and engage with other scientists and nonscientific audiences and collaborators through our intensive formalized training in communicating science and community engagement (i.e., the Community Mentor Program). Programmatic outcomes include the development of training modules and workshops on the most effective community engagement and communications strategies exportable to the national CTSA consortium.

NIH Research Projects · FY 2024 · 2024-07

Modified Project Summary/Abstract Section Type 2 diabetes (T2D) and metabolic dysfunction-associated fatty liver disease (MAFLD) are two highly-associated metabolic disorders. It is well-documented that African Americans are 60% more likely to develop T2D than non-Hispanic whites. Multiple factors contribute to these disparities, including biological and clinical factors, as well as health system and social factors. However, available evidence of environmental factors are not enough to explain this significant T2D disparities between African Americans and non-Hispanic whites. Our database mining identified a sequence variant (rs142619613) within the coding region of YY1 gene (Yin Yang 1). rs142619613 occurs mainly in African Americans. This nucleotide variant (G-C) leads to Glu47Asp missense mutation, which significantly increased stability and transcriptional activity of YY1. We further established YY1 as a transcription activator of miR-23b/27b/24. MiR-23b/27b/24 selectively drove AKT phosphorylation but prevented phosphorylation of FOXO1, thereby promoting de novo lipogenesis (DNL) and gluconeogenesis. Phenotypically, antagonizing miR-23b/27b/24 alleviated hepatic insulin resistance, hepatosteatosis, hyperglycemia and MASH. Based on these findings, we hypothesize that rs142619613, by augmenting the YY1-miR-23b/27b/24 axis, selectively drives phosphorylation of AKT and prevents phosphorylation of FOXO1, thereby shifting the metabolic program of the liver towards increased gluconeogenesis and DNL and exacerbating hepatic insulin resistance, hyperglycemia and hepatosteatosis. The objective of this R56 project is to elucidate the role of miR-23b/27b/24 in promoting MASH development and evaluate the effect of rs142619613 on lipid and glucose homeostasis in human hepatocytes. Our long-time goal is to explain the higher incidence of T2D and MAFLD in African Americans and develop YY1 and miR-23b/27b/24 as potential therapeutic targets against both conditions. Two specific aims are designed to test our hypothesis in this R56 project. In Aim 1, we will establish the role of miR-23b/27b/24 in promoting hepatic inflammation and fibrosis and evaluate the impacts of miR-23b/27b/24 knockdown and liver-specific expression of Wwc1 on food intake, body weight, glucose homeostasis, and lipid homeostasis. Completion of this aim will establish gain-of miR-23b/27b/24 in promoting MASH. In Aim 2, we will assess the role of rs142619613 in disrupting lipid and glucose homeostasis in human hepatocytes. Understanding how T2D and MAFLD develops is of critical importance to design therapeutic strategies to combat both chronic illness. The results will provide novel insights into the mechanisms of the higher prevalence of T2D in African Americans, which may lead to rational therapeutic strategies for T2D.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMARY / ABSTRACT In general, enzymes are very precise at catalyzing a specific canonical reaction that fits within a particular metabolic network. Still, no enzyme is a perfect catalyst. The inherent flexibility of proteins makes it difficult for enzymes to distinguish their canonical substrate from structurally related compounds. Thus, many enzymes act on unintended substrates (i.e., substrate promiscuity). Substrate promiscuities result in the formation of unintended or damaged metabolites (i.e., metabolite damage) that can be a useless drain on metabolism, and may be inhibitory and/or reactive, sometimes leading to toxicity. Accordingly, metabolite damage repair enzymes exist for the specific purpose of counteracting metabolite damage, often by converting a damaged metabolite to a canonical one. The physiological importance of metabolite damage and its repair has been revealed over the past ~15 years as a handful of metabolic diseases in humans were discovered to be caused by disruption of metabolite damage repair genes, many of which are highly conserved across the three domains of life. The proposed project will address metabolite damage repair associated to the TCA cycle – a universal core metabolic pathway that is involved in energy conversion and is a source of chemical building blocks that supplies much of metabolism. The TCA cycle is a hotspot for metabolite damage due to high carbon flux through the pathway and chemical intermediates that are structurally similar organic acids, which can engage in promiscuous side reactions catalyzed by the abundant cycle enzymes. I have identified and characterized several highly conserved metabolite damage control systems related to vitamin, cofactor, and amino acid metabolism, and my training has empowered me with a unique skillset and perspective that is allowing me to make similar discoveries related to the TCA cycle. One enzyme that I have identified is particularly intriguing. A prevalent side-reaction of the TCA cycle enzyme succinate dehydrogenase oxidizes malate to enol-oxaloacetate (OAA), a metabolically inactive form of OAA that is a potent inhibitor of the TCA cycle. Our results provide strong evidence this side reaction is one of the most prevalent promiscuous reactions in nature, and that enol-OAA is a potent inhibitor of the TCA cycle. We identified a universally conserved enzyme, OAT1, that removes the inhibitor, and show that bacterial cells lacking OAT1 have a severely attenuated TCA cycle. The proposed work will integrate biochemical, genetic, and metabolomics/ fluxomics approaches to determine how OAT1 impacts the physiological and metabolic states of prokaryotic and single and multicellular eukaryotic model organisms. Completing this project will lead to the detailed characterization of a previously unrecognized but critical aspect of the TCA cycle, ultimately redefining one of the most universal core metabolic pathways in biology. This work will also provide insights into mitochondrial metabolism and physiology that will impact human health and disease, and deliver a metabolite damage repair enzyme for use in optimizing synthetic biology platforms.

NIH Research Projects · FY 2026 · 2024-06

A. Project Summary / Abstract The ongoing monkeypox (mpox) outbreak, with a total of more than 87,000 cases reported in over 100 non- endemic countries, highlights the pandemic potential of poxviruses. Due to its zoonotic nature with broad hosts and unidentified natural reservoirs of the causative agent, mpox virus (MPXV), future spillovers and outbreaks are expected even if the current outbreak is contained. Although typically manifesting in milder symptoms than smallpox, MPXV infections can still cause significant morbidity and mortality (up to 10% for clade I) that need to be therapeutically mitigated. The clinical efficacy of FDA-approved smallpox antiviral drugs is unclear or unpromising for treating MPXV, necessitating dedicated and expanded efforts in developing MPXV antivirals. The goal of this grant application is to develop novel antivirals for treating MPXV. Prior research from our team has developed and validated an efficient and robust reporter assay using the vaccinia virus (VACV), and identified and characterized three high quality and chemically distinct antiviral hits with strong potencies (EC50 = 0.14‒2.1 M; plaque reduction at 10 M 1,400‒160,000-fold), no cytotoxicity (CC50 > 250 M) and largely favorable ADME properties. The immediate focus of this grant is to further develop these three hit series into pre-clinical candidates using a highly integrated, multi-disciplinary approach combining medicinal chemistry, virology, ADME / pharmacokinetic (PK), toxicity, animal efficacy and proteomics. Three Specific Aims are proposed: 1) optimization and expansion of the identified hits. Iterative structure-activity relationship (SAR) and structure-property relationship (SPR) studies will be carried out via extensive analog synthesis, antiviral assay and ADME profiling to obtain optimized leads. Hit expansion will also be conducted by screening the high quality synthetic compounds in PI’s lab as well as preselected commercial compounds; Aim 2) in vivo PK, toxicity and efficacy studies, where the acute and sub-acute toxicity and PK studies in mice will guide compound selection for efficacy studies in mice; Aim 3) determine the antiviral mechanism of action (MOA). The molecular targets and antiviral MOA of optimized leads will be determined using a few different methods: a) identify the viral replication stage(s) targeted by the leads; b) select resistant mutants by passaging virus in cultured cells in the presence of individual lead compound, and sequence viral genomes to identify affected genes; The compound will then be tested in recombinant viruses harboring the selected mutations; c) design and synthesize chemical probes to label, pull down, and identify the target protein(s); d) test metal chelating analogs in resolvase biochemical assays. Overall, 2-3 antiviral leads with strong potency, favorable PK profiles, minimal toxicities, and established MOA will be developed, setting the stage for further pre-clinical development of MPXV drugs that will also contribute substantially to countering other orthopoxviruses.

NIH Research Projects · FY 2026 · 2024-06

The Carlson Group applies the diverse tools of chemical biology to understand how bacteria harness their limited genome to inhabit nearly every ecological niche on our planet. These “simple” single-celled organisms are remarkable in their ability to respond to and survive in the presence of diverse environmental stressors. This is largely accomplished through related but specialized proteins that are activated under disparate conditions. With NIH support, the PI has been investigating two protein families that are crucial to a bacteria’s ability to respond to its environment and maintain cell wall integrity, histidine kinases (HKs) and penicillin-binding proteins (PBPs). The PBPs are required for cell wall construction and are the targets of b-lactams, the most used class of antibiotics in the world. The HKs sense changes in the periplasmic space and transmit this information inside of the cell, allowing bacteria to respond to fluctuating environments. The HKs are also important in infection and antibiotic resistance. Clearly, a deeper understanding of when, where, and how the PBPs and HKs are functionally differentiated and used to enable cell survival will be crucial in combating the rapidly approaching “post-antibiotic era.” Chemical probes are an essential tool for the characterization of these differentiating factors. We have pioneered the development of activity-based probes (ABPs) for the PBP and HK families that specifically label catalytically active enzymes, yielding a read-out of their functional state. These molecules are required to investigate how functionally related proteins differ from one another in their catalytic activation, substrate or ligand recognition, localization, and biomolecular interactions. The overall vision for this MIRA project is to build on the PI’s foundation of results and leadership in this area and apply a chemical approach to expand the understanding of these two enzyme families. We will achieve this vision by expanding our library of PBP-selective ABPs and applying these chemical probes to investigation of the roles of these proteins in cell growth and division, as well as adaptation to environmental perturbations. We will also optimize the chemical probes and internalization agents needed to deliver ABPs into the bacterial cytoplasm, enabling global mapping of HK activation upon exposure to stimuli. Finally, we will validate the HKs as viable targets for the development of anti-virulence agents and adjuvants and optimize the potency of our lead compounds. Our combined expertise in chemical synthesis, ABPs, mass spectrometry-based -omics, biochemistry, and microbiology make us uniquely qualified to untangle the web of apparent functional redundancy within these enzyme families. We also have long-standing collaborators with expertise in the HKs and PBPs, as well as methods critical to this project. The overall impact of this work will be a drastically increased and rigorously tested understanding of cell wall biosynthesis and signal transduction in bacteria and a suite of freely accessible research tools. Ultimately, the amassed knowledge and tools will enable us to understand and predict how a signal is propagated into bacterial action and to hijack the involved proteins for disease treatment.

NIH Research Projects · FY 2026 · 2024-06

Project Summary The number of children born to opioid-dependent mothers has increased over 300% in the past two decades. As abstinence from opioids during pregnancy is not recommended, opioid medication replacement therapy, such as methadone, represents the standard of care for pregnant women with opioid use disorder thus perpetuating the birth of these opioid-exposed babies. Previous work has shown that prenatal opioid exposure predisposes for future substance misuse. Given its widespread accessibility, alcohol is one of the most likely addictive sub- stances that these prenatal opioid-exposed children will encounter. Due to the significant morbidity and mortality associated with excessive alcohol drinking, it is critically important to understand how prenatal methadone ex- posure (PME) predisposes these children for future problematic alcohol use and how alcohol interacts with PME to produce differential behavioral responses to alcohol. To elucidate mechanisms related to how PME may pro- duce enhanced alcohol-related behaviors, we developed and validated a mouse model of PME. Our model re- capitulates many clinical features of prenatal opioid exposure, including producing neonatal opioid withdrawal. Using our model, we find that PME increases alcohol drinking only in males, consistent with many clinical and preclinical studies that show that males are more severely impacted by prenatal opioid exposure. Given the role of the dorsal striatum brain region in modulating many aspects of alcohol drinking, we biochemically explored the proteome of the dorsal striatum and found that PME had a greater effect on protein and protein phosphory- lation expression in males than females, consistent with our drinking data. Pathway analyses of our proteomics data implicated glutamate and long-term synaptic depression plasticity (LTD) in the dorsolateral striatum (DLS) as being disrupted by PME. We further discovered that PME reduced dorsal striatal glutamate transmission and disrupted LTD. Recent work from our laboratory demonstrates that alcohol induces glutamatergic synaptic plas- ticity and disrupts LTD at anterior insular cortex inputs to the DLS (AICDLS synapses) in mice, but only in male mice and this was associated with enhanced male alcohol drinking behavior. We reasoned that the male-specific, PME-induced increase in binge-like alcohol consumption may utilize similar mechanisms as the male-specific, alcohol-induced AICDLS synaptic changes that govern excessive alcohol consumption. We hypothesize that PME produces synaptic adaptations exclusively in males that enhance AICDLS glutamatergic transmission that in turn govern the elevated binge-like alcohol consumption seen in male, but not female PME mice. In this project we will use a multidisciplinary approach combining home-cage binge drinking with brain slice electro- physiology, dorsal striatal cell type reporter mice, quantitative synaptic proteomics, ultrastructural expansion mi- croscopy, and wireless in vivo optogenetic manipulations of synaptic transmission. This project will provide crit- ical neural mechanistic knowledge for deciphering how PME enhances future alcohol drinking behavior in males and why females may be resistant or resilient to the effects of PME.

NIH Research Projects · FY 2026 · 2024-06

Project Summary/Abstract Adolescent conduct problems, such as aggression, defiance, violence, and criminality, represent a major public health concern with substantial costs to individuals, their families, and larger society. Current evidence-based programming addressing adolescent conduct problems is resource intensive and has limited effectiveness. The proposed R34 study will investigate the feasibility of using a mindfulness-based intervention (MBI) as a new approach to preventing escalations in conduct problems among at-risk youth in a school setting. MBIs have been demonstrated to support the development of effective self-control, a key risk factor underlying adolescent conduct problems. However, MBIs have not previously been evaluated for their effectiveness in preventing the development of serious conduct problems. Notably, previous implementations of MBIs with adolescents have been limited by a lack of generalization of mindfulness skills in daily life, particularly during times of stress. The proposed study will enhance an existing evidence-based MBI, Learning to BREATHE (L2B), with a recently-developed mobile app designed to support greater generalization of skills in daily life and times of stress. The proposed study will be completed in two primary phases, including Phase 1, a small pilot feasibility- focused 3-arm RCT will assign youth (n=120) to either the standard L2B intervention, L2B+App, or Skills for Success, an active control life skills intervention. We will address four primary aims. Aim 1 will focus on refining the implementation (n=10) of L2B enhanced by the mobile app (i.e., L2B+App), and Phase 2, in which a L2B+App intervention, including app usability, app functioning and features, integration with face-to-face programming, and cultural responsiveness of the integrated app. Focus groups and facilitator feedback from the Phase 1 pilot implementation will support refinement prior to the Phase 2 feasibility RCT. Further refinement will be supported by additional participant focus groups and facilitator interviews in Phase 2. Aim 2 will evaluate the feasibility of research procedures in the Phase 2 3-arm RCT, including our ability to reach recruitment, enrollment, and retention of participants, randomization procedures, and data collection procedures acceptability, appropriateness, engagement, feasibility, and fidelity of delivery i.e., L2B, L2B+App, and Skills for Success) with a focus on achieving benchmarks in each area. Finally, Aim 4 involves using a mixed-methods approach to refine all protocols, including facilitator training, intervention implementation, and research procedures, in preparation for a subsequent fully-powered efficacy trial. Successful completion of these aims will provide critical preliminary data required for the efficacy trial. benchmarks in . Aim 3 will evaluate the of the three interventions ( Moreover, the proposed research will demonstrate the potential of app- based strategies for enhancing delivery of mindfulness skills in at-risk youth. This program of research has the potential to innovate our approach to preventing adolescent conduct problems through the incorporation of mindfulness techniques as well as reduce the high costs and burden associated with these behaviors.

NIH Research Projects · FY 2026 · 2024-06

Project Summary The bacterial genome is organized into a compact form, known as the nucleoid, consisting primarily of the supercoiled circular DNA chromosome and DNA-binding proteins. Generation and maintenance of the nucleoid structure, which arises without a nuclear membrane or the chromatin present in eukaryotes, is critical to bacterial health; understanding the factors that give rise to this structure are thus important for promoting synergistic bacteria and disrupting antagonistic pathogens. Likewise, the dynamics governing DNA loci diffusion and intranucleoid transport govern the time scales for DNA-protein binding. A key open question surrounding the structure and dynamics of the nucleoid is elucidating those factors that can be attributed to its polymeric nature and those that are of biological origin. Indeed, many of the most intriguing ideas emerging from the microbiology community in the past decade, such as density waves, the glass-like behavior of the nucleoid in the absence of metabolic activity, and the connection between solvent quality and macrodomain formation, rely on connecting nucleoid biology to polymer physics. Unfortunately, making these connections in a definitive, quantitative manner has been frustrated by (i) the lack of a suitable physical model of the nucleoid and (ii) experimental data that, when combined with such a model, are sufficient to establish the governing biophysical principles. To resolve this gap in our understanding, this proposal couples (i) a multiscale, polymer physics-based model of the nucleoid that reflects both its equilibrium and non-equilibrium aspects to (ii) state-of-the-art biophysical experiments probing the nucleoid structure and dynamics. Any model of the nucleoid must respect the biological fact that the nucleoid is not a fully equilibrated system; the combination of topological domains, macrodomains and chromosome interacting domains are evidence that the nucleoid is not in a global equilibrium that samples the full configurational space, even if these domains can locally equilibrate over the time scale of cell division. We will parameterize a computationally tractable, coarse-grained (CG) model of the entire out-of- equilibrium nucleoid based on simulations of the rapidly equilibrated, fine-scale phenomena. By construction, the CG model will incorporate large-scale, non-equilibrium constraints imposed by biology. We will then study nucleoid structure and dynamics over parameter ranges far beyond those available in existing data sets, using both GPU-accelerated simulations of the CG model, live cell imaging, and a microfluidic experimental system. The experiments will improve the CG model through feedback between them, while the CG model will reveal the most useful systems for experimentation and aid in interpreting their results. Our approach will definitively establish the physical laws governing nucleoid structure and dynamics. Applying those laws at biologically relevant parameter ranges then provides a foundation for identifying when polymer physics can (or, even more importantly, cannot) be used to understand nucleoid biology.

NIH Research Projects · FY 2026 · 2024-06

Summary Different cell types, each with unique composition, properties, and functions, are essential to sustain life in an organism. Bulk analysis of such heterogeneous mixtures of cells does not distinguish the characteristics of a given cell type or differences between cells within the same type, missing information that is key to understanding molecular mechanisms foundational to the rational design of therapies for disease and ailments. The goal of this proposal is to develop single-cell and imaging methods using metal-bearing probes to monitor metabolic processes in live cells such as siRNA delivery, protein synthesis, and post-translational modification. The cells will then be fixed and processed for subsequent detection of protein and mRNA markers. There are three aims: (1) Delivering molecular probes to viable cells to explore molecular pathways in single cells by CyTOF. (2) Barcoding prenylation probes to explore molecular pathways in mixed samples of single cells. (3) Exploring the effect of aging in murine primary cells by CyTOF and murine tissue by MIBI-ToF. The proposed work will expand the use of multi- parametric capabilities of mass cytometry (i.e. cytometry by time-of-flight, CyTOF, and multiplexed ion beam imaging by time-of-flight, MIBI-ToF) in the biotechnological and biomedical fields. The development of these methodologies will explore the causal relationship between autophagy and senescence in murine liver models, exploring the hypothesis that the mevalonate pathway provides a link between the two. The ultimate goal is to understand the relationship between autophagy, mevalonate, and senescence pathways in aging and inform emerging treatments to counteract the detrimental effects of senescent cells.

NIH Research Projects · FY 2026 · 2024-06

People with severe mental illness (SMI) are ~3x more likely to contract and ~7x more likely to die from vaccine-preventable disease. And yet, vaccine uptake is significantly lower among people with SMI, likely due to greater vaccine delay/refusal despite availability (“hesitancy”). Critical knowledge gaps – including extremely limited knowledge of whether and how unique features of SMI modulate hesitancy – impede selection of interventions on vaccination for people with SMI. This K01’s purpose is to determine unique reasons for hesitancy among people with SMI (Aim 1), reveal causal pathways to hesitancy in people with SMI and forecast which variables in these pathways are likely the best intervention targets (Aim 2), and examine the impact of education about herd immunity – which improves vaccine intentions in the general population – on vaccine intentions and uptake in people with SMI (Aim 3). In Study 1, we will recruit people with SMI and two comparison groups: people with depression/anxiety and healthy people. Participants will complete measures of SARS-CoV-2 and influenza vaccine intentions/uptake, of factors that may be uniquely relevant to hesitancy in people with SMI, and of variables implicated in general population models of vaccine hesitancy. We will compare groups’ willingness to vaccinate, their level of concern about vaccines’ dangerousness, and their endorsement of anti-vaccine information (Aim 1). We will use active-learning causal discovery algorithms and intervention calculus to reveal causal pathways to hesitancy in people with SMI, quantify the lower-bound total causal effect of variables in these pathways on hesitancy, and prescribe future experiments to resolve any uncertainties in these causal pathways (Aim 2). In Study 2, we will use established natural language processing tools to analyze rates of and attitudes to anti-vaccine misinformation in Tweets from Study 1 participants. This analysis will complement our laboratory assessments of anti-vaccine misinformation endorsement for Aim 1. In Study 3, we will recruit a subset of participants from Study 1’s healthy and SMI groups. We will compare the effects of education about herd immunity across these groups, with the goal of identifying differential efficacy and examining the need to tailor existing interventions to people with SMI (Aim 3). Our rigorous characterization of unique aspects of vaccine hesitancy among people with SMI, along with causal pathways we reveal, our corresponding predictions about the impact of manipulating variables in these pathways, and our quantification of herd immunity education’s effect on vaccine decisions, will provide critical road-maps for empirically-informed interventions on hesitancy in people with mental illness. These road-maps will move us closer to a world where effective, evidence-based interventions on hesitancy are deployed to protect people with SMI from vaccine-preventable disease.

NIH Research Projects · FY 2025 · 2024-06

SUMMARY Immuno-oncology studies continue to grow which seek new therapies leveraging immunogenic, non-normal peptide sequences (neoantigens) arising from tumor-specific alterations at the genomic, transcriptomic or proteomic level. Non-normal DNA and RNA sequences that may encode neoantigens can be identified from next-generation sequencing (NGS) data, and further prioritized by their predicted binding to the class I or II major histocompatibility complex (MHC) as an indicator of immunogenicity. Immunopeptidomic enrichment of the peptide-MHC complex coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) can confirm the existence of predicted neoantigens as well as other tumor-associated antigens (TAAs) derived from normal protein sequences, including those with post-translational modifications (PTMs). This powerful approach requires `immunopeptidogenomic' informatics tools that integrate NGS and MS data analysis. Despite steadily growing numbers of cancer researchers pursuing these studies, they lack a centralized informatics resource tailored to these informatics requirements. As a solution, we will develop the immunopeptidogenomic (iPepGen) informatics resource for immuno-oncology research. iPepGen will leverage the Galaxy bioinformatics ecosystem, offering cancer researchers accessible workflows to predict neoantigens from NGS data and confirm their presence from MS-based immunopeptidomics data, including training resources housed in the Galaxy Training Network to promote community adoption. We will achieve our goals through these Specific Aims: Aim 1: Optimize and harden modular workflows for identifying, prioritizing and curating neoantigen candidates detected from genomic and/or transcriptomic alterations; Aim 2: Optimize and harden state-of-the-art MS-based immunopeptidomic analysis modules for identifying and verifying MHC-bound neoantigen and TAA peptides; Aim 3: Disseminate tested and optimized workflows and engage in training activities to promote community adoption of the iPepGen resource. Our team brings complementary, world-class expertise necessary for success in developing the iPepGen resource. PIs Griffin and Jagtap have developed widely used Galaxy-based multi-omic tools and training materials for cancer research. PI Nesvizhskii is a world-leader in development of computational tools for quantitative, MS-based proteomic and peptidomic analysis. Development, testing and optimization of tools, workflows and training materials will be guided by collaboration with cancer researchers conducting Driving Immuno-oncology Projects (DIPs). The iPepGen resource will offer a critically needed resource to advance game-changing immunotherapy studies impacting a wide-variety of cancer types.

NIH Research Projects · FY 2025 · 2024-05

Project Summary / Abstract Vocal prosody inflections are essential to function in everyday communication. For example, pitch (F0) – conveys information about expressing agreement or disagreement, to correct, to signal novel information, or to indicate between-the-lines meaning. Cochlear implants are notoriously poor at delivering accurate information about a talker’s voice pitch (Carlyon et al. 2002; Oxenham 2008). Difficulty in perceiving prosodic information is highlighted by CI patient reports of communication breakdown and listening complaints where the tone or hidden meaning of a message is misunderstood (Rapport et al. 2020). Predictably, standard speech perception measures based on intelligibility alone do not explain variability in communication ability or quality of life for listeners with cochlear implants (Capretta & Moberly 2016; Hinderink et al. 2000; Moberly et al 2018). Conversely, test batteries that specifically include skills related to vocal prosody are better at predicting quality of life and communication-related measures among listeners with cochlear implants, underscoring the need for further understanding of how to assess vocal prosody perception (Luo et al. 2018; Panzeri et al. 2021). To provide a person with the best ability to function in a social world where perception of a talker’s tone and intention play a vital role, we must better understand how perception of prosody is affected by the use of a cochlear implant. The first aim of the proposed work will compare perception of prosodic focus by listeners with normal hearing and cochlear implants. Importantly, listeners will report prosody perception for whole-sentence stimuli and provide responses using a multiple sliding-scale interface that will allow for differentiation of stronger and weaker perceptions of prosody, as well as characterization of errors in prosody perception relative to the talker’s intended target. The second aim will explore the perceptual weighting of pitch, intensity, and duration cues in prosody perception for both groups, as well as basic psychophysical measures to determine perceptual access to prosodic prominence. The third aim will examine the impact of noise on prosodic cue weighting by these two listener groups. Results from these examinations will show how differences in prosodic cue weighting result in weaker prosody perception and frequent mistakes for CI listeners, as well as a greater impact of noise on prosody perception. The long-term goals of this project are to improve our understanding of prosody perception for CI users. These goals directly address the NIDCD’s priority of increasing our knowledge on communication by individuals with hearing loss in real world environments. The training program involves extensive instruction in behavioral measures, acoustic analysis and manipulation, and advanced statistical analysis relevant to the proposed research. This research will be conducted under the mentorship of leading experts in hearing at an institution with a strong track record of hearing loss research, a history of training grants, and interdisciplinary collaboration in clinical sensory science.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Reward neurocircuitry relies in part on endogenous neuropeptide signaling to integrate complex, multivalent information and influence behavior. Opioid addiction can result in neuropeptide imbalances in the nucleus accumbens (NAc), a brain region critical for processing reward. Signaling by neuropeptides derived from VGF (non-acronymic) may be dysregulated by exogenous opioid exposure with repeated withdrawal. The neuropeptide precursor VGF and its derived peptide TLQP-62 serve important functions in experience- dependent plasticity in the nervous system, and VGF transcripts are upregulated in the nucleus accumbens (NAc) after drug exposure and withdrawal. My preliminary data demonstrate that knocking out VGF in the NAc eliminates the behavioral changes typically evoked by opioid exposure and withdrawal in mice. Because these behavioral adaptations are proxies for neuroplasticity in reward circuitry, we hypothesize that VGF is necessary for synaptic plasticity in the NAc. The objective of this work is to interrogate the function of VGF and its derived peptide TLQP-62 in facilitating synaptic plasticity in the NAc, and to understand its contribution to withdrawal- evoked behaviors. First, I will measure the effect of exogenous TLQP-62 on synaptic transmission in the NAc using whole-cell patch-clamp electrophysiology in acute mouse brain slices. Past work in other regions of the central nervous system have shown that TLQP-62 potentiates excitatory transmission. We therefore hypothesize that TLQP-62 will augment excitatory glutamatergic transmission into the NAc. I will then use RNA in situ hybridization to map endogenous VGF transcripts in the NAc and identify in which neuronal subpopulations it is expressed. Finally, I will evaluate the effect of endogenous VGF on withdrawal-evoked behavior adaptations. I will combine transgenic mice and viral vectors to conditionally knock out VGF expression from the NAc and use a model of spontaneous oxycodone withdrawal before performing a battery of behavioral assays to determine the locomotor, social, and sensory impact of VGF in the NAc. With these methods, spanning physiology, neuroanatomical mapping, and behavior, we will build an understanding of the function of VGF and TLQP-62 in the NAc and the role they play in opioid exposure and withdrawal. Beyond identifying a novel target for treating opioid use and withdrawal, the interdisciplinary techniques used in this proposal will examine the basic science mechanisms of clinically urgent questions, laying the groundwork for a career planted at the intersection of clinical need and basic research.

NIH Research Projects · FY 2026 · 2024-05

This is a new R25 proposal entitled the “Minnesota-Tuskegee Surgery Training Experience leading to Posters and Presentations” (M-STEPP). M-STEPP is a 9-week summer research experience for up to 10 undergraduate students from under-represented groups in biomedical sciences in partnership with Tuskegee University (TU). The program will provide translational research experiences related to diseases impacting the cardiovascular system with a clear deliverable of a poster, presentation, or a brief research report. This approach leverages an interdisciplinary partnership among faculty across the University of Minnesota (UMN) who have funded research portfolios within the scope of NHLBI’s mission. Mentors have research programs studying: 1) inflammation and endothelial cell biology; 2) atherosclerotic disease; 3) cardiovascular transplantation; 4) CVD risk factors related to diet, exercise, neuromodulation, and bariatric surgery; 5) development of CVD across the lifespan; 6) smoking as a driver of disease; 7) regenerative approaches to increase myocardial function; 8) the microbiome and its connection to metabolic disease; and 9) hemorrhagic shock and cardiovascular collapse following trauma. We use turnkey resources in the Department of Surgery to expose trainees to preclinical research, including Experimental Surgical Services, The Visible Heart Lab, and the Pre-Clinical Research Center, all of which have experience in the training of undergraduates from underrepresented groups. Our approach is based on an ongoing partnership between TU and UMN, now in year 2, with the primary stated goal of increasing the pipeline into science careers of those underrepresented in medicine. The focus of this R25 proposal is for scholars to complete a health services research project with exposure to a bench research experience. Interested scholars will be invited back for a second summer to deepen their exposure and expertise either in health services research (e.g., use of natural language processing) or a discrete project in translational benchwork. Our objectives are to introduce students to: 1) clinical experiences that will stimulate questions and long-term interest in cardiovascular health and the recognition of gaps in knowledge and care; 2) health services research through hands-on approaches; 3) data analysis; 4) principles of scientific writing with preparation of an abstract to scientific meetings; 5) speaking at the Department of Surgery Research Week; 6) critically evaluating scientific research in journal clubs; 7) clinical exposure and critical thinking with focused exposure to the simulation center and basic life support training. To ensure that the program is responsive to mentees, curriculum will be adapted based on pre- and post-completion surveys as well as formal evaluations of skills with a pre- and post-test, and social media and graduate school guidance resources will be leveraged to maintain mentee trajectory and monitor program success. This approach catalyzes the use of proposed resources to under-represented minorities to facilitate careers in biomedical research.

NIH Research Projects · FY 2026 · 2024-04

Abstract Direct intravenous delivery of gasses has been always considered an absolute contraindication due to the fear of air embolism. However, if it were made possible, direct intravenous high content oxygen solutions could ameliorate injury, improve tissue hypoxia and buy time for definitive interventions and/or prevent invasive interventions such intubations. The basic resting minute oxygen requirements of a human are 3ml/kg/min or 200- 300 ml of O2/min. During CPR, blood flow is significantly decreased and oxygenation at the lungs is frequently severely impaired. Both those conditions lead to vital organ and whole-body hypoxia. An effort to increase the overall oxygen content of circulating blood could have significant effect on end-organ oxygenation even with lower blood flow and could significant delay irreversible injury that leads to significant morbidity and eventually mortality in this patient population. We have developed a method with which a highly pressured mixture of saline and oxygen can be delivered directly into the vein at 1 Atm in order to dramatically increase the circulating oxygen blood content. We utilize high pressures (50-100 Atm) at room temperature (25-28C) to dissolve large volumes of oxygen in the infusate to make a clinically relevant therapy and sustain body oxygen requirements for up to 10- 20 min/L of saline infused. The concept is based on the simple idea that when the highly pressurized solution (eg. at 50 atm and 25C the dissolved volume of O2 /saline is ~2.5/1) is released under constant pressure through a nozzle that controls bubble nucleation rate and bubble size distribution evolution, a “mist” consisting of nucleated nanobubbles and saline water molecules can be delivered, which can be directly mixed with the returning venous blood. The presence of venous, unsaturated hemoglobin acts as an absorption sink for the delivered O2nanobubble solution (O2NBS). An optimal rate of mixing can be safe and minimize bubble formations through the process of immediate O2 uptake. Residual nanobubbles do not coalesce and therefore could circulate uninterrupted through the circulatory system to act as an additional oxygen reservoir. In Aim 1 we will study the physics and engineering methods to optimize the solution and nanobubble size characteristics under different upstream Pr/nozzle conditions. We aim to minimize nanobubble average size to safely deliver it in saline at 1atm and 37C. Aims 2 and 3 will assess the effect of the intravenous oxygen solution on blood and tissue oxygenation in animal models of simulated cardiac arrest and during prolonged CPR. Finally, a large 72 -hour survival study will be performed. The implications to public health are self-evident and far-reaching for diseases like cardiac arrest, hypoxic respiratory failure, emergencies requiring intubation and anesthesia.

NIH Research Projects · FY 2025 · 2024-04

The Academy of Behavioral Medicine Research (ABMR) is guided by its mission to advance the field of behavioral medicine by creating and disseminating knowledge, cultivating discourse, and inspiring change that culminates in better health for all. ABMR members are distinguished mid-level and senior scientists, including MDs and PhDs, elected by their peers for contributions to behavioral medicine research. Our mission primarily is accomplished through the annual scientific meeting that brings together our diverse membership and invited thought leaders to share the latest scientific advances, exchange cutting-edge ideas, and provide stimulating and engaging discussions in an informal but rigorously scientific atmosphere. ABMR scientists lead extensive NIH-supported programs of research that integrate biomedical, behavioral, and social sciences to improve the health and well-being across the lifespan. Our members focus on understanding biologic mechanisms of aging that underlie leading causes of death and disability (heart disease, cancer, Alzheimer's, diabetes) and developing and implementing effective interventions to maintain health and reduce the burden of age-related chronic conditions. In 2021, we added an early-stage investigators (ESI) program to the ABMR annual meeting, specifically to foster career development and leadership skills of promising behavioral medicine scholars with their first K- or R-level award from NIH. This R13 conference grant will support the planned 2024 and 2025 annual scientific meetings, including participation of 6 invited Keynote Speakers (3 per year) and our ESI Program (10 per year). The 2024 meeting will be held June 20-23, 2024, at The Depot in Minneapolis, MN and the 2025 meeting will be held June 25-28, 2025 in Portland, ME. Our R13 has 3 Aims. In Aim 1, we seek to promote interdisciplinary discourse on the latest advances in behavioral medicine, public health and health policy to promote healthy aging and reduce chronic disease disparities. Within this overall theme, the 2024 meeting will focus on strengthening the impact of our science by enhancing collaborations among scientists, discussing cutting-edge and integrative team science approaches to research. The 2025 focus will be on strengthening the impact of the science by enhancing collaborations outside our traditional academic boundaries, discussing innovative partnerships and the critical value of two-way communications with local communities, healthcare systems, and policymakers. Both meetings will include Keynotes, symposia, roundtable discussions, working groups, and ample time for networking. In Aims 2 and 3, we will support diverse and up-and-coming ESIs through the ABMR ESI Fellowship Award, providing a leadership workshop, mentorship and career development training focused on strategies for sustaining and strengthening independence and leadership. This R13 enables ABMR to nurture the success of the next generation of behavioral medicine researchers and leaders, which aligns with NIH’s Next Generation Researchers Initiative, and enables ABMR to advance the field of behavioral medicine as it has since its inception 45 years ago.

NIH Research Projects · FY 2026 · 2024-04

Project Summary: Despite reductions in antimicrobial use and implementation of antibiotic stewardship programs, antimicrobial resistant (AMR) pathogens continue to emerge and persist, causing millions of deaths every year. The long-term goal of this proposal is to decrease clinical AMR incidence by advancing knowledge of how AMR develops, persists and transmits to pathogens within the microbiome. The overall objective of this project is to develop a novel high-throughput method for generating enriched long-read sequencing data that accurately reflects the in situ dynamics of AMR genes across entire microbial communities. This is a major gap in methodology that hampers knowledge discovery and leads to suboptimal patient outcomes. The rationale is that this method will enable fundamental advances in both applied and basic research through: improved prediction about when and how bacteria exchange AMR genes; and rapid, lower-cost, higher-throughput generation of metagenomic resistome data. The project objective is achieved through three specific aims: 1) Increase the sensitivity and specificity of probe-based pre-sequencing enrichment; 2) Generate contextualized metagenomic sequence data with real-time enrichment of antimicrobial resistance genes; and 3) Evaluate the performance of triple-enrichment using samples with diverse antimicrobial resistance gene composition. Under aim 1, probes for all potential off- target (i.e., AMR) DNA will be designed using a novel compressed colored de Bruijn graph. Probes will be manufactured and used to deplete off-target DNA from metagenomic samples prior to enrichment of AMR genes. For aim 2, adaptive Nanopore sequencing will perform real-time rejection of off-target DNA using a compressed pangenomic index of all off-target regions, followed by on-the-fly classification of antimicrobial resistance genes. For aim 3, the potential bias of probe- and sequence-based enrichment will be quantified on a gene-by-gene basis to fully evaluate the performance of the enrichment workflow for various sample types and use cases. The proposed research is innovative for two major reasons: first, it enables the applied use of microbial ecology for clinical and public health applications, which is a major gap in the field of metagenomics; second, it allows for deep sequencing and robust genomic co-localization of rare and low-abundance metagenomic targets, which is currently very difficult to obtain. The proposed methodology will open new horizons for basic research into how and why bacterial communities evolve; and under what conditions diverse bacteria share critical functions such as AMR. The proposed research is significant because it will have broad applicability to many critical activities for combating AMR, including patient-centric decisions about antibiotic use and public health activities such as AMR surveillance. The proposed advancements will help predict AMR emergence events in patients and populations; identify hot-spots of AMR development and dissemination in hospitals and the community; and help track critically important AMR genes that may be rare or low-abundance within a given environment or set of samples.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are a novel class of medications that reduce CVD events, particularly heart failure (HF) outcomes in patients with HF and reduced or normal left ventricular (LV) function. The mechanism for the effect of SGLT2i is not well understood, but they may beneficially alter LV structure and function through changes in plasma proteins. Atrial myopathy is a novel biomarker for increased cardiovascular disease (CVD) risk. Atrial myopathy is characterized by abnormalities in the structure, function, and electrical conduction of the left atrium (LA). LA function can be quantified using strain analysis with 2-dimensional echocardiography (2DE) allowing detection of LA dysfunction before overt structural changes and LA enlargement occur. LA dysfunction is associated with increased risk for atrial fibrillation, stroke, dementia, and heart failure. Despite this risk, there are no proven therapies to treat or prevent LA dysfunction. Untargeted proteomic analyses done in adults with LA dysfunction are suggestive of an association with plasma proteins involved in cardiomyocyte function and inflammatory pathways. The goal of this K23 Career Development Award is to conduct a 9-month double-blind placebo-controlled RCT of empagliflozin (an SGLT2i) in 80 individuals at risk for HF but no diagnosis of HF or diabetes. Individuals will be identified by screening patients who visit the Cardiology clinic at the University of Minnesota. The primary endpoints for the proposed RCT include changes in LA and LV function assessed by 2DE at baseline and 9 months; and change in 8 a priori identified plasma proteins. Aim 1 will test the hypothesis that change in LA function from baseline to 9 months will be more favorable in the empagliflozin group than placebo. Aim 2 will test the hypothesis that change in LV diastolic from baseline to 9 months will be more favorable in the empagliflozin group than placebo. Aim 3 will test the hypothesis that compared with placebo, empagliflozin will be associated with greater change in the selected proteins after 9 months. The proposed training and mentoring plan has the following goals: 1) acquire expertise in clinical trial design, implementation, and analysis; 2) develop expert knowledge in left atrial myopathy; 3) acquire necessary skills to design, execute, and interpret studies involving proteomic analysis; 4) develop enhanced collaborative skills, written and oral communication skills, and management skills necessary to lead a multidisciplinary team. The training plan includes attendance at educational seminars and scientific meetings, structured mentoring, formal coursework, and hands on experience on this mentored K23 project and other collaborative research with mentors. In summary, the requested K23 support will provide the training and mentoring necessary for the candidate to establish himself as a successful independent physician scientist in the field of preventive cardiology. The proposed research will also provide novel preliminary evidence regarding the effect of empagliflozin on LA and LV function in an understudied population, informing a future R01-funded RCT evaluating SGLT2i treatment effect on CVD outcomes in patients without diabetes or HF.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Background. Agents that inhibit Androgen Receptor (AR) signaling are the standard-of-care treatment for prostate cancer (PC) patients. Unfortunately, essentially all patients develop resistance to AR targeting therapies (ART). We previously found robust B7-H3 (CD276) expression in metastatic PC patients resistant to ART. In our recent phase II clinical trial, we found that B7-H3 targeting agents improved clinical outcomes for 66% of PC patients for reasons unknown. We separately examined mechanistic regulators of B7-H3 expression with computational and laboratory approaches. We preliminary found that B7-H3 expression was regulated by AR and its co-regulators (FOXA1, HOXB13) at four transcription regulatory sites. Altogether, we hypothesize that AR activity regulates B7-H3 expression and thus response to B7-H3 targeted therapies. Specific Aims and Study Design. We propose to determine mechanisms of B7-H3 expression by examining the necessity of all B7-H3 transcription factors and regulator sites in PC (AIM 1). This includes the AR co- regulators HOXB13 and FOXA1, as well as the four B7-H3 enhancer sites that were hyperactivated in metastatic PC compared to primary disease. We will also determine how androgens or ARTs, which are used clinically, impact the susceptibility of ART resistant tumor cells towards B7-H3 targeted therapies (AIM 2). We will utilize single cell RNA-sequencing platforms to identify the genes and signaling programs associated with response to B7-H3 therapies in single cells. Lastly, we aim to identify the clinical and molecular characteristics of the PC patients with predicted response to B7-H3 targeted therapies (AIM 3). We will use computer- modeling tools to characterize the somatic changes and clinical features of patients that previously responded to a B7-H3 targeted therapy. We will then apply this response signature to ~7000 additional tumors to identify significant clinical and molecular features that are predictive of response. Anticipated Outcomes. We anticipate that the candidate transcription factors and regulatory sites of B7-H3 will control B7-H3 expression in PC tumors. Further, such changes in cell surface B7-H3 expression will greatly impact the tumor-forming capacity and ART resistance. We also anticipate that clinical forms of androgens or ART will alter B7-H3 expression and thus impact response to the B7-H3 targeted therapies. Lastly, we expect to identify many of the additional clinical and molecular features associated with B7-H3 expressing tumors. Future Directions and Impact. We anticipate this study will uncover mechanistic insights of B7-H3 expression and a further understanding of PC patient response to B7-H3 targeted therapies. We shall also identify the mechanisms by which B7-H3 regulates ART resistance, tumor formation, and cell signaling. Lastly, our findings will impact patient stratification methods in the upcoming B7-H3 clinical trials, which will be essential to the development and deployment of future B7-H3 therapeutics in PC patients.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract More than three-fifths of Americans with mental health disorders (MHD) have not received treatment in the prior year. Societally, under-utilization of mental health care has a substantial impact on disability and contributes to premature mortality through causes such as suicide. The problem of low mental health care use persists despite progress in increasing the rate of health insurance coverage. A major barrier to mental health services is cost, even among the majority of Americans who have either employer-sponsored or individual market commercial health insurance. As such policy interventions to reduce costs of mental health services among people with commercial insurance might have substantial effects on mental health services use and potentially quality. In 2021, New Mexico passed SB 317 “No Behavioral Health Cost Sharing,” which prohibited cost-sharing for mental health services in the commercial insurance plans that the state regulates. This law was the first of its kind nationally, and represents a stark natural experiment in reducing cost-related barriers to mental health services. The proposed study will assess the implementation and effects of the SB 317 No Behavioral Health Cost Sharing law with four specific aims. 1) Characterize the implementation of SB 317 from the perspective of clinicians and patients in New Mexico who are directly affected by the law, using qualitative interviews with key stakeholders. 2) Assess the effect of SB317 on MHD services benefit generosity using insurance documents that summarize plan benefits. 3) Evaluate the effect of SB317 on use of MHD services using quasi-experimental methods and comprehensive commercial claims data. 4) Evaluate the effect of SB317 on the quality of MHD services received. Understanding the way that zero cost-sharing affects mental health services will provide important new evidence to help guide mental health financing decisions to most- efficiently improve mental health services use.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY Substance use disorders (SUDs) impair the health and well-being of over 40 million Americans and claim 106,000 lives each year due to a lack of effective treatments. The nucleus accumbens (NAc), which consists of medium spiny neurons (MSNs) marked by expression of Dopamine-1 or -2 receptors (D1 or D2, respectively), is a reward center of the brain that exhibits changes in synaptic plasticity that drive addiction. D1-MSNs drive rewarding behaviors, such as addiction, while D2-MSNs drive aversive behaviors. Thus, identifying precise molecular targets that modulate D1-MSN synaptic signaling would hold promising therapeutic potential for SUDs. Angiotensin converting enzyme (ACE) is selectively expressed in D1-MSNs within the NAc. In the brain, ACE hydrolyzes numerous neuropeptides, including the enkephalin Met-enkephalin-Arg-Phe (MERF), which has a high affinity for opioid receptors. ACE inhibition (ACEi) increases levels of MERF in the NAc, which acts in a - opioid receptor (MOR) dependent manner at excitatory presynaptic inputs to drive long-term depression (ACEi- LTD) specifically onto D1-MSNs. However, it has not yet been determined which specific excitatory and inhibitory presynaptic inputs express ACEi-LTD, as NAc MSNs receive a diverse array of long-range excitatory and local inhibitory inputs. The main goal of this proposal is to determine which specific excitatory and inhibitory presynaptic inputs to NAc MSNs express ACEi-LTD, revealing the circuit-specific synaptic plasticity mechanisms evoked by ACEi which is critical for developing circuit-informed therapies for SUDs. Towards this, excitatory opsins will be expressed virally in an input and cell type-specific manner in individual inputs to the NAc. Whole-cell patch-clamp electrophysiological recordings will be taken from D1- and D2-MSNs while optogenetically stimulating individual excitatory or inhibitory inputs. Optically- and electrically-evoked postsynaptic currents will be measured before, during, and after application of an ACE inhibitor to determine which specific excitatory and inhibitory inputs exhibit ACEi-LTD. The MOR-dependence of ACEi-LTD will be determined using pharmacological and genetic blockade of MOR signaling. Since thalamic inputs to the NAc are known to drive opiate dependence, and since prior studies suggest that ACEi modulates cortical excitatory inputs to the NAc, Aim 1 will investigate the ACEi sensitivity of thalamic and cortical excitatory inputs onto MSNs. Additionally, since fast-spiking interneurons (FSIs) provide the most robust regulation of MSN output, and since low-threshold spiking interneurons (LTSIs) are known to decrease presynaptic release upon MOR-agonist treatment, Aim 2 will investigate the ACEi sensitivity of FSI and LTSI inhibitory inputs onto MSNs. This proposal will reveal the circuit-specific effects of ACEi, which selectively targets D1-MSNs of the NAc that drive reward and contribute to addictive behaviors. Since ACEi attenuates fentanyl preference in mice, ACE may be a precise and safe therapeutic target for the prevention and/or treatment of SUDs such as opioid use disorder.

NIH Research Projects · FY 2026 · 2024-04

MRI is the most advanced and the most used tool in our armamentarium to study the human brain. However, highest possible resolution achievable with existing methods falls short of goals set for the next generation Human Connectome Project (HCP) or the BRAIN Initiative that calls for the capability to study the organizing principles of the human cortex at the mesoscopic or sub-mesoscopic scale. Our long-term goal is to advance ultrahigh field (UHF) MRI through innovative technical developments applied at the highest possible magnetic field available for human studies. The overall objective of this study is to develop a new suite of MRI tools that, when synergistically combined with the unique UHF of 10.5 Tesla (10.5T) and optimized instrumentation (RF coils and gradients), will enable us to acquire high-quality, whole- and partial-brain functional MRI (fMRI) at unparalleled spatiotemporal resolutions, ushering in the next generation HCP and approaching the ambitious resolution target (i.e., 0.01-µL voxel volumes) set in the strategic plan developed by the second BRAIN Initiative Working Group. The overall objective will be accomplished by pursuing four specific aims: Under aim 1, we will develop and optimize new high-channel-count transmit and receive RF arrays to leverage SNR gains available at 10.5T while enabling RF parallel transmission (pTx) for flip angle homogenization and SAR reduction. For aim 2, we will devise two new pTx pulse design frameworks that incorporate RF-power-related constraints for water- selective excitation tailored for whole-brain scan without the need for additional fat saturation, and a fat-selective saturation and a water-selective excitation, suitable for partial-brain scan of a single imaging slab. Under aim 3, we will develop i) a comprehensive pTx-enabled 3D GRE EPI sequence for rapid high-quality high-resolution whole-brain and partial-brain scans, all with joint motion and field correction, ii) a computational toolkit enabling real-time motion detection, background field measurement, and GIRF-based field dynamics determination using NMR field probes, and iii) a tailored image reconstruction for producing motion- and field-corrected images based on field dynamics and motion detection. The results will be compared to those obtained with our long-standing and extensively used 2D simultaneous multi-slice EPI sequence. For aim 4, we will develop and demonstrate brain fMRI at spatiotemporal resolutions that are much higher than what is possible with existing methods. The research proposed in this application is significant because it is expected to have a broad positive impact on various fields including ultrahigh-field MRI, RF hardware, high-resolution fMRI, parallel transmission, data acquisition, image reconstruction, and neuroscience. The proposed research is innovative because it represents a new and substantive departure from the existing approaches by shifting focus to developing a synergy of various technical innovations that can help realize the full potential of 10.5T, enabling ultrahigh-resolution fMRI and increasing our ability to study brain function at the mesoscopic scale.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Chronic kidney disease (CKD) is highly prevalent, affecting approximately 37 million U.S. adults. CKD- mineral and bone disorder (CKD-MBD) is a common comorbidity of CKD that results in increased risk of cardiovascular and bone disease and associated morbidity and mortality. Abnormal phosphorus (P) metabolism is central to the development of CKD-MBD and is intimately linked with calcium (Ca) metabolism. Incomplete understanding of the underlying physiology of Ca and P in CKD and in response to treatments is a major knowledge gap that hinders research and clinical progress in CKD-MBD. P and Ca physiology is complex and involves interacting effects of three regulatory hormones, calcitriol, parathyroid hormone, and fibroblast growth factor-23, on a multi-tissue axis of intestine, kidney, and bone. Despite the complexity, most human research has relied on serum and urine Ca and P measures to infer aspects of whole-body physiology. Yet, prior work has shown that serum and urine Ca and P are not reliable markers of whole-body balance or intestinal absorption in CKD. Formal metabolic balance studies combined with isotope tracers can reveal the underlying whole-body Ca and P physiology and, notably, can distinguish intestinal absorption from bone resorption and formation. This project seeks to fill this knowledge gap through two specific aims. Aim 1 will determine the effects of dietary P restriction on P and Ca intestinal absorption, whole-body balance, and kinetics in adults with moderate CKD. Aim 2 will determine the effects of calcitriol, a key P and Ca regulatory hormone, on P and Ca intestinal absorption, whole-body balance, and kinetics in adults with moderate CKD. Each aim will be addressed in a clinical study using a two-phase randomized cross-over design with controlled feeding and metabolic balance and kinetics methods in adults with moderate-stage CKD. In Study 1 (Aim 1), subjects will be randomly assigned to a cross-over order of low and high diet P, achieved through manipulation of P source based on current understanding that inorganic P sources have much higher bioaccessibility compared with P found naturally in plant and animal foods, as recommended by current guidelines. In the second study (Aim 2), subjects will be randomly assigned to a cross-over order of calcitriol and identical placebo. Each study will consist of a 1-week run-in period on the controlled diet/intervention, 1-week full metabolic balance period with complete urine and stool collections and oral and intravenous administration of P and Ca isotopes for kinetic modeling to determine components of P and Ca metabolism including: intestinal absorption, renal clearance, and movement to and from bone, with kinetic measures continuing during a third week. After a washout period, subjects will cross-over to the second intervention period. The long-term objective of this project is to advance foundational knowledge of whole-body P and Ca physiology in CKD that will contribute to progress in translational and clinical research with the goal of reducing morbidity and mortality associated with CKD-MBD. This relates to the mission of the NIDDK to support research aimed at improving the health and quality of life of patients with kidney disease.

NIH Research Projects · FY 2026 · 2024-04

The PRKACA gene encodes for the catalytic subunit of protein kinase A (PKA-C). Recent ge- nomic efforts have identified several single-site mutations, insertions, and aberrant fusions directly linked to endocrine diseases. Specifically, a few somatic or germline mutations/insertions have been linked to adenoma-associated Cushing’s syndrome and its associated cardiac myxomas. Additionally, two chimeras are drivers for fibrolamellar hepatocellular carcinoma (FLHCC) and intraductal oncocytic papillary neoplasms (IOPNs) of the pancreas and bile ducts. Recent litera- ture and our preliminary studies suggest dysfunctional spatiotemporal regulation and loss in substrate specificity of PKA-C may represent common traits for the progression of these dis- eases. However, there are no conclusive molecular mechanisms underlying the progression of these diseases. We hypothesize that these PKA-C variants disrupt the allosteric cooperativity that drives the assembly/disassembly of the regulatory complexes, unleashing active PKA-C. We pro- pose to use state-of-the-art liquid-state NMR techniques coupled with biophysical and biochemi- cal approaches to characterize the interactions between PKA-C and its regulatory subunits and the endogenous regulator PKI. We will assess how mutations, insertions, and fusion chimeras of PKA-C disrupt intra- and inter-molecular allostery, leading to aberrant regulation and disease. Understanding the regulatory mechanism of PKA help determine the molecular etiology of these diseases and aid in the future design of strategies to precisely control the aberrant activity of kinases.