Mayo Clinic Rochester

NIH Research Projects · FY 2025 · 2025-09

The rationale of the Mayo Clinic Polycystic Kidney Disease Research Core Center (MPRC) is to provide reagents and data through the PKD-Research Resource Consortium (PKD-RRC) to the PKD community to facilitate breakthroughs in PKD research. The operational philosophy of the MPRC is to provide resources that can be transformative to the PKD community in areas of perceived need. The MPRC builds on a long history of PKD research at Mayo and uses the expertise of its faculty in genomics, radiological imaging, and gene editing to generate these reagents. The Center also builds upon biological resources and generated reagents including a large, well characterized cohort of autosomal dominant PKD (ADPKD) patients, a PKD and polycystic liver disease biobank, ADPKD patient DNA variant data, and developed radiological software to characterize PKD kidneys and liver. The MPRC has a single Biomedical Research Core, focused on the study of human ADPKD: The Human ADPKD Population and Materials (APM) Core. In addition, there is a Research Development Core (RDC) and an Administrative Core that oversees the operations of the MPRC, manages communications within the Center and with the PKD-RRC, and organizes the education and training part of the Center, including the Summer Student Enrichment Program. The APM Core focuses on providing deep, longitudinal phenotyping data, including radiological imaging images and genetic results from 1250 Mayo ADPKD patients, to enable investigators to better understand disease progression. Additionally, the Core will collect and distribute PKD serum, urine and kidney and liver tissue, to query pathogenesis. The RDC highlights Mayo expertise in vivo gene editing and imaging software development that can be further developed for breakthrough technologies. Overall, the MPRC will provide much needed reagents, tools, and expertise to transform PKD research in the US.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Ulcerative colitis (UC) affects nearly one million US adults. Refractory disease is still common, and the current trial-and-error treatment approach causes delays in identifying optimal therapy for individual patients. Anti-TNF remains a commonly prescribed biologic for UC, but approximately one-third of patients fail to respond and about one-third lose response over time. Understanding the role of IBD-associated genes in intestinal epithelial cell (IEC) injury, especially in determining therapy response or resistance, is a critical unmet need for aligning patients with effective treatment options. Our long-term goal is to develop cytoprotective strategies and identify formalin-fixed, paraffin embedded (FFPE) tissue signatures that correlate with therapy response. The objective of this proposal is to elucidate IEC injury pathways that govern anti-TNF therapy effectiveness, and to discover predictive biomarkers in FFPE biopsies indicative of anti-TNF treatment outcome. Our rationale is that TNF- dependent and independent mechanisms of IEC injury dictate anti-TNF therapy response or resistance. Our novel approach employs an innovative mouse model, A20/Abin-1T-ΔIEC, exhibiting severe colitis that mirrors anti-TNF refractory UC. This model is instrumental in identifying TNF-independent pathways of intestinal injury. Complementing this mouse model, we will apply pioneering spatial transcriptomics assays to analyze archived FFPE patient biopsies, revealing treatment response signatures in routinely collected samples. Predictive biomarkers in FFPE biopsies would integrate best with current clinical workflows, since nearly all IBD patients have archived FFPE biopsies. Additionally, we leverage well-characterized colonic organoids derived from healthy controls and UC patients for mechanistic in vitro assays to characterize primary IEC responses to death stimuli. The specific aims include: 1. Delineating TNF-independent (anti-TNF refractory) mechanisms of IEC death via the A20/Abin-1T-ΔIEC mouse model, hypothesizing that IL-1B signaling is a crucial driver of mucosal injury. 2. Validating IEC injury pathways correlated with anti-TNF treatment response and non-response in FFPE specimens, using a customized gene panel for detailed spatial transcriptomics. 3. Assessing differential sensitivities of UC-derived IEC organoids to death stimuli and cytoprotective factors, hypothesizing a distinct response pattern in UC-derived organoids compared to controls. This proposal integrates a novel mouse model, advanced spatial transcriptomics, and patient-derived organoid cultures to pinpoint critical mechanisms of IEC injury and develop biomarkers indicative of anti-TNF treatment response. By advancing this research, we aim to significantly impact precision medicine for UC, aligning medications to individual patient epithelial biology, while identifying new therapeutic targets.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT Inflammatory bowel disease (IBD) is characterized by inconsistent response to current therapies and persistent activation of pathogenic effector CD4+ T cells, implying that regulatory T cells (Tregs) are potentially dysfunctional. Recent evidence suggests that inflammatory-like Tregs are present in the intestinal lesions of individuals suffering from refractory IBD; however, the underlying mechanism and its contribution to IBD are poorly understood. Thus, the OVERALL OBJECTIVE of this proposal is to aid in the generation of more preliminary data for a planned R01 application that will explore the molecular mechanisms by which lipid metabolites enforce Treg phenotype and immunosuppressive function during chronic intestinal inflammation. These insights will facilitate the development of therapeutic strategies for manipulating Tregs to treat IBD. Untargeted metabolomics revealed that certain sphingolipids accumulate in human Tregs. This proposal aims to then explore how these lipids are synthesized and elucidate the importance of these lipids to metabolic homeostasis, cell signaling, and Treg function. Building on the unbiased metabolomics data, we also found that the abundance of these lipids in Tregs can be modulated by physiologically and pathophysiologically relevant signals. Therefore, we hope to explore the functionality of Tregs depleted of these lipids in the setting of experimental mouse models of chronic intestinal inflammation to establish the function of these lipids in Tregs in a manner that mirrors human IBD. Based on these observations, we formulated the CENTRAL HYPOTHESIS that long-chain ceramides promote metabolic homeostasis and resistance against the acquisition of effector-like phenotype in Tregs, resulting in the preservation of immunosuppressive function. The following independent SPECIFIC AIMS are designed to test two integrated hypotheses. First, we will directly test the hypothesis that ceramides prevent excessive oxidation of fatty acids linked to the glycolytic-lipogenic metabolism and effector IL-17-producing T helper (Th)17 cell-like phenotype. Furthermore, we will test the hypothesis that ceramides inhibit the activation of the nutrient-sensing signaling pathway associated with effector type T helper (Th1) cell-like phenotype. Second, we will test the hypothesis that Tregs depleted of ceramides exhibit poor resolution of IBD in an animal model. We propose to utilize sophisticated approaches relevant to health and IBD pathophysiology to test our central hypothesis. This proposal, which is technically and conceptually innovative, is also significant because it presents a novel concept in Treg biology and identifies new mechanisms for therapeutically optimizing Tregs to halt the refractory IBD.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Registries are by far the most important source of data for orthopedics, particularly joint replacement. The importance of registries surpasses even randomized clinical trials (RCT) due to the long duration of postoperative surveillance, the substantial costs associated with the procedures and implants under investigation, and the heterogeneity that exists across institutions, regions, and countries. Unfortunately, building a large-scale and comprehensive registry is difficult. On one hand, comprehensive datasets usually result from expensive small cohort projects such as the Osteoarthritis Initiative (OAI). Universal implementation of this model is not feasible as smaller institutions might not have the resources to employ the required personnel or build the extensive initial infrastructure. On the other hand, nationwide registries such as Medicare databases are comparatively sparse and unhelpful. In orthopedics, large-scale registries like the American Joint Replacement Registry (AJRR) face challenges of data contribution. Participation is currently voluntary and the fractionated nature of the US healthcare system limits the data quality of contributions. Thus, the balancing act in registry construction is between comprehensive depth and participation/completeness. If data points are too onerous to abstract, participation will be low; if completeness is prioritized, interesting data points are difficult to include. To solve both these problems and take the next step in national-scale registry construction, we will develop automatic methods of data abstraction. The potential time and cost benefits of an entirely automated abstraction pipeline are immense, in addition to allowing for registries to easily scale to accommodate the records produced by the millions of arthroplasties performed nationwide. Our central hypothesis is that large language models, with proper fine-tuning, grounding, and prompting, can acquire trustworthy orthopedic-specific performance enabling them to interpret clinical notes for data extraction and complex synthesis tasks. Successful completion of this aim will yield fine-tuned LLMs capable of 1) efficiently and accurately extracting critical data for automated orthopedic registry curation, and 2) interpreting clinical notes for patient-specific phenotyping. These advancements are anticipated to reduce barriers to clinical registry construction and increase the comprehensive depth of registry data, encouraging cross-institutional collaborations on significant health issues. Additionally, increased registry participation will facilitate the integration of pragmatic and nested RCTs within registries, enabling prospective data collection and generation of high-level evidence to refine surgical techniques and implant design, ultimately improving patient outcomes.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT As the primary non-invasive imaging tool in cardiology, echocardiography delays and misdiagnoses significantly affect outcomes for the 5.5% of the U.S. population with cardiac issues. Automation of echocardiography interpretation is proposed but remains limited to models with narrow image analysis functions. Research into Visual-Linguistic Models (VLMs) and Large Language Models (LLMs) to build an Artificial Intelligence-enabled Echocardiography Interpretation System (AEIS) presents a solution to this problem and paves the way to next- generation Generalist Medical AI (GMAI) systems. In this proposal, Dr. Chieh-Ju Chao is seeking a K08 Mentored Career Development Award to advance his training to develop and evaluate an AEIS. The mentoring team is led by Drs. Bradley Erickson (primary mentor) and Fei-Fei Li (co-primary mentor), both are worldclass experts with complementary AI ability in medical imaging and computer vision. They are funded by agencies including NIH, NSF, and DoD, with a proven record of accomplishment of successfully mentoring junior investigators. The Research Training Plan will equip Dr. Chao with advanced skills through mentorship from a cross-site, multidisciplinary team, with Dr. Bradley Erickson serving as the central figure to ensure training quality. In Aim 1. Dr. Chao will train with Drs. Fei-Fei Li and Ehan Adeli to pretrain VLMs using PubMed Open-Access figures and the CheXpert Plus datasets, followed by fine-tuning with Mayo's echocardiography data to enhance caption accuracy. Aim 2. involves training with Dr. Curtis Langlotz to optimize LLMs for factual correctness and expert preferences, and Dr. Erickson for quantitative hallucination detection to build an LLM system for echocardiography report summarization. In Aim 3, Dr. Chao will train with Dr. Imon Banerjee on multi-modal framework organization and evaluation to assemble the AEIS, and with Dr. Jae Oh on designing clinical reader studies, and Dr. Garvan Kane on AI-based workflow optimization to study the effectiveness and human-AI interaction of the AEIS. Dr. Chao has a solid foundation in echocardiography and AI, 13 publications in the medical AI field, including 6 as a first or co-first author since he entered in 2021. The Career Development Plan will enhance Dr. Chao's ability on advanced AI research skills for developing and evaluating an AEIS, with career skills gained through practical experience and coursework. The environment at Mayo Clinic, enhanced by resources from Stanford's Institute of Human-centered Artificial Intelligence through our collaborative program, is outstanding for innovative research. Excellent infrastructure (datasets, cloud and local computing resources and other equipment) is available. In summary, the robust mentoring team and training plan are expected to thoroughly prepare Dr. Chao for an independent career centered on developing and evaluating future GMAI systems in cardiovascular medicine, to improve efficiency and quality of echocardiography interpretation, as well as patient outcomes and longevity.

NIH Research Projects · FY 2025 · 2025-09

The aging population has led to an increase in cognitive impairment (CI) such as mild cognitive impairment or dementia. As the aging population grows, the need for effective diagnostic, management, and predictive tools is more urgent. With this landscape, there is great need for precision medicine approaches considering individual factors, such as sex, in the understanding and management of dementia. In particular, women have a disproportionate burden of dementia. However, the examination of women-specific differences has not been well integrated into precision medicine approaches, despite the extensive literature demonstrating sex differences in brain structure and function over the lifespan. Women-specific factors earlier in life, such as menopause, menopausal hormone therapy, pregnancy, and hypertensive disorders during pregnancy, are hypothesized to affect cognitive health and dementia. Some studies have identified a stronger association between biomarkers, such as apolipoprotein E (APOE) ε4 genotype, and dementia or cognitive decline in women compared to men. Additionally, the progression of dementia in women can also be uniquely influenced by a myriad of biological, social, and environmental variables. Despite the high prevalence of dementia in women, disparities in diagnosis, access to healthcare, clinical trials, treatment, and care management have been reported. Electronic health records (EHRs), particularly clinical free text, contain invaluable information about patients’ medical history to track longitudinal health conditions. Yet, EHRs have not been fully utilized to systematically capture or distinguish women-specific health events, trends, or patterns that could be invaluable in understanding CI progression. To address this gap, we will analyze women-specific health patterns from EHRs that can address diagnostic disparities in CI progression. We will study these questions using: (a) the population-based Mayo Clinic Study of Aging cohort (n=6,531) with longitudinal cognitive assessments and extensive clinical characterization, providing an ideal gold standard for CI; and (b) the Rochester Epidemiology Project, which links longitudinal EHRs for a large Midwestern population (~1.4 million people in 27 counties). Additionally, we will use nationwide data from National Alzheimer’s Coordination Center to increase representation of the population and to enhance the generalizability. The primary goal of this project is to develop an informatics tool to extract women’s health conditions from multi-site EHRs (Aim 1), analyze differences in CI symptoms, diagnosis, and progression in women in relation to biomarkers (Aim 2), and develop fair and explainable predictive models for CI, assessing the prognostic value of women’s health indicators to guide targeted health adjustments (Aim 3). The proposed tools and methodologies will address diagnostic and treatment disparities in women, ensuring transparent and fair healthcare decisions. The proposed aims align closely with the imperative need and could significantly impact the lives of women at risk for Alzheimer’s disease and other dementias.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Alcohol-associated liver disease (ALD) and metabolic dysfunction-associated steatotic liver disease (MASLD) are leading causes of liver transplantation, morbidity, and mortality. With the obesity epidemic and increasing alcohol consumption, a mixed-etiology disease, metabolic syndrome-associated alcoholic liver disease (MetALD), has emerged. Accurate diagnosis and risk prediction of ALD, MASLD, and MetALD are currently hindered by the lack of distinct histological features. To address this critical knowledge gap and its impact on diagnostic accuracy and the development of precision medicine, this proposal introduces an innovative approach that harnesses the power of multi-omics data integration using machine learning (ML) techniques. Our central hypothesis is that each disease has distinctive features identifiable by ML and spatial transcriptomics, enabling the development of new diagnostic and prognostic tools. The rationale for this is that current histological methods lack specific features, whereas ML has shown potential for enhanced diagnostic accuracy. In our preliminary data: 1) we developed several ML tools, including one that distinguishes between ALD and MASLD with high accuracy, another that assigns zones in liver biopsy whole slide images (WSI), and identified a morphometric signature for severity in ALD; 2) we identified a unique zonal gene signature profile for MetALD from a limited spatial transcriptomic dataset. Building on these preliminary successes, our proposal outlines two specific aims designed to further dissect the complexities of steatotic liver disease: 1. We will define the etiology and predict the severity of steatohepatitis using novel ML tools as follows: a. Classify the etiology as ALD, MetALD, or MASLD on WSI based on ML. b. Predict disease severity using a graph neural network model for comprehensive morphometric analysis. 2. We will identify distinct profiles that confer prognostic value for MetALD as follows: a. Define zonal and unique regions of interest pathways of liver injury using spatial transcriptomics b. Develop a multimodal prognostic algorithm for MetALD based on the histological features of the graph neural network, clinical and serological findings, and spatial transcriptomics. This proposal is technically and conceptually innovative, as it seeks to integrate histological features with ML for diagnostic precision and disease severity prediction, advancing our understanding of the underlying pathways of injury in MetALD. These outcomes will guide the development of noninvasive diagnostic and prognostic markers, aligning with NIAAA’s mission to improve alcohol-related liver disease outcomes. This proposal ensures that I achieve my goal of becoming an independent physician-scientist specializing in ML, spatial transcriptomics, and liver diseases, through the advanced training and dedicated research time provided by the K08 and the well-rounded mentorship and tremendous resources provided by the Mayo Clinic.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Deep Brain Stimulation (DBS) is a prevalent therapy approved by the U.S. Food and Drug Administration that uses electrical stimulation of brain nuclei to treat the symptoms associated with movement disorders. However, a lack of immediate feedback to determine its efficacy for treating other disorders has led to suboptimal therapeutic effects. Thus, re-imagining DBS may lead to more effective therapies that treat the root cause of the disorder, and not just address its symptoms. These therapies may require exploring innovative approaches that leverage our understanding of neural plasticity mechanisms. Spike-timing dependent plasticity (STDP) is a well characterized phenomenon that can induce neuroplasticity via phase-locked activation of pre- and post-synaptic neural elements. Here, we propose a novel approach that combines STDP and DBS (STDP-DBS) and is capable of selectively changing maladaptive plasticity associated with pathologic neural activity and behavior observed across multiple neurologic and psychiatric disorders, including alcohol addiction. In this proposal, STDP-DBS applied with precise timing at two regions of the brain such as the Globus Pallidus internus (GPi) and the Anterior Cingulate Cortex (ACC) will enable selective bidirectional gain control of cortico- striato-pallidal signaling. In turn, this will allow control of maladaptive neural signaling that contributes to the pathology of neurologic disease while avoiding off-target disruption of non-pathological brain activity, in turn minimizing potential side effects of conventional DBS. The work proposed here represents the first steps toward correcting maladaptive plasticity associated with neurologic and psychiatric disorders. In turn, this may enable a new avenue for treatment of a wide range of disorders such as addiction, Tourette’s syndrome, obsessive compulsive disorder, schizophrenia, Parkinson’s disease, major depressive disorder, and many others.

NIH Research Projects · FY 2025 · 2025-09

(PLEASE KEEP IN WORD, DO NOT PDF) Telomere biology disorders (TBD) are multisystem disorders caused by premature shortening of telomere lengths. Hematopoietic stem and progenitor cells (HSPC) in TBD show impaired proliferative capacity manifesting as bone marrow failure (BMF). We and others have shown that HSPCs from TBD patients may develop somatic mosaic events that lead to context-specific clonal hematopoiesis (CH), overcoming replicative constraints. Due to TBD-specific unique stressors, these patients have a higher prevalence of CH (40% vs. 6% in age-matched non-TBD individuals) and a characteristic context-specific mutational spectrum mainly in DNA damage response and repair (DDRR) pathways and the p.S34 position in the spliceosome gene U2AF1, which results in multiple missplicing events. U2AF1MT-S34 is associated with rapid clonal expansion and evolution to myeloid neoplasm in non-TBD population, but TBD-U2AF1MT-S34 patients show a perplexing stability over time. This suggests that clonal dynamics and clinical outcomes of CH in TBD differ from age-related CH, with the underlying mechanisms behind these differences still unknown. We hypothesize that U2AF1MT-S34 somatic events result in splicing alterations in key pathways that rescue hematopoiesis in the context of TBD and may lead to adaptive and/or maladaptive outcomes, including progression to myeloid neoplasm. To test this, we will create and characterize a new K562 hematopoietic stem cell line to study our preliminary observations in TBD patients with U2AF1MT-S34 CH. In Aim 1, we will use a novel CRISPR/Cas9 method developed in our laboratory to create a K562 TERTMT cell line. We have used this method successfully in HEK-293t cells resulting in a phenotype that resembles TBD (low telomerase activity, short telomeres, replicative impairment). Using the same approach, we will introduce the U2AF1MT-S34 variant in this K562 TERTMT cell line and characterize the effect of these co-occurring mutations on telomere length, replicative capacity and morphology. In Aim 2, we will characterize the transcriptomic missplicing changes exerted by U2AF1MT-S34 in relevant pathways (DDRR, splicing, and telomere maintenance) in the context of short telomeres using long-read RNA sequencing (Oxford Nanopore). This project offers both conceptual and methodological innovation by creating a new model of TBD-CH and using it to characterize the downstream effect of the most frequent somatic event observed in these patients.

NIH Research Projects · FY 2025 · 2025-09

Hepatobiliary and pancreatic (HPB) cancers are a group of rapidly fatal malignancies with poorly understood etiology and biology. These cancers are often diagnosed at a late stage, precluding curative therapy. To address the increasing problem of higher incidence and mortality of HPB cancers in Africa, there is a pressing need to (1) identify and empower cancer researchers and clinicians with the most current knowledge and research skills in the field to contribute meaningfully to understanding the etiology and prognostic factors associated with these cancers, and (2) help develop sustainable, practical actions to reduce the incidence and mortality of these cancers in Africa. There is also an urgent need to harness our collective expertise and resources in the United States with African investigators to advance the science of HPB cancers in Africa. This proposal is a renewal of a successful three-year conference series of the Africa HepatoPancreatoBiliary Cancer Consortium, as supported by R13CA265018. These conferences include hands-on skills workshops in surgery, advanced endoscopy, ultrasound, pathology, and patient advocacy, followed by plenary and discussion sessions on the pathobiology of hepatopancreatobiliary cancers, scientific writing, mentorship, designing joint research projects among investigators in Africa and the United States, prevention and early detection, clinical care in low-resource settings, project management training for research coordinators, and training in new laboratory techniques/technology, among other topics. The past three years have led to the initiation of multi-national collaborative research projects, clinical training opportunities, and mentorship programs. The funding of this R13 grant will allow us to harness the momentum we have created to further these existing efforts. At the conclusion of the two-year R13 grant, we will have enhanced the expertise in Africa and the United States to address the increasing mortality rates from hepatobiliary and pancreatic cancers. Through this project, we will continue to develop a vibrant and collaborative group of investigators equipped with appropriate skills to meaningfully contribute to the advancement of the science of HPB cancers. The results of our research collaborations will not only improve our understanding and treatment of HPB cancer etiology and biology in Africans, but also of all persons in the United States.

NIH Research Projects · FY 2025 · 2025-09

Despite well-established benefits of reduced mortality and improved quality of life, <25% of eligible patients participate in cardiac rehabilitation (CR). Disparities in CR participation are particularly worse among patients most affected by cardiovascular (CV) disease including those of lower socioeconomic status and those living in rural and dense urban areas. Participation barriers include inflexible CR hours and considerable distances to CR centers. In the post-COVID-19 pandemic era, novel CR delivery methods to mitigate barriers to care are crucial. Data-driven telehealth models using virtual technologies have emerged to expand CR access through home-based delivery. One such modality, virtual worlds (VWs), are 3-D, immersive computer-based environments allowing users to interact via online personas (avatars), simulate in-person experiences, and social network. VWs are applied for physical rehabilitation, health education, and chronic disease management and could address CR participation barriers. Our major project goal is to use a Hybrid Type 1 implementation trial design to rigorously test the efficacy of a behavioral theory-informed, 12-week, VW-based CR (VWCR) intervention compared to center-based CR (CBCR) for improving CV health and CR participation among cardiac patients. CV health will be measured by the American Heart Association Life’s Essential 8 (LE8) composite score, an evidence-based metric of 8 health-promoting behaviors/clinical factors (e.g., diet, blood pressure) that improves CV outcomes. We will conduct a multiphase, multicenter, 2-arm, randomized controlled non-inferiority trial with 150 adult cardiac patients. We hypothesize that patients randomized to our patient-centric VWCR intervention will have noninferior CV health profiles and higher adherence rates than those randomized to CBCR. This R01 proposal aligns with the NLM’s focus on next-generation technologies to “reach more people in more ways through enhanced dissemination and engagement.” Building on our patient informed VWCR preliminary work, we propose 2 aims. Aim 1 will determine the effect of VWCR on CV health among cardiac patients compared to CBCR. Primary outcome is the LE8 score. Aim 2 will determine whether patients randomized to VWCR will have improved participation in and adherence to CR compared to those in CBCR. Primary outcome is adherence (attendance of ≥70% of sessions prescribed). We will use mixed methods to assess adherence and sustainability/scalability potential. Secondary outcomes include major adverse CV events, psychosocial measures (quality of life, self-efficacy, self-regulation, social support), cost effectiveness and implementation outcomes according to the RE-AIM framework. A Patient/Community/Stakeholder Advisory Board will provide input for all activities. Our innovative, reproducible VWCR intervention integrates theory-informed and empirically supported components to influence the LE8. If successful, our results can pave the way for scalable, broad implementation of VWCR to increase CR accessibility and improve CV health outcomes among cardiac patients.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT Expanding pharmacotherapeutic options and finding treatment response biomarkers for existing medications are both important for facilitating personalized medicine in alcohol use disorder (AUD). Accordingly, NIAAA advocates for research exploring the neurobiological mechanisms of AUD and recovery. Acamprosate, an FDA-approved antidipsotropic, has been suggested to modulate various neurotransmission systems including glutamate and GABA, yet its exact efficacy mechanisms remain speculative. Neuroactive steroids, including allopregnanolone (ALLO), play crucial roles in modulating GABAergic inhibition by binding to GABAA receptors. Given the disruption of GABAergic neurotransmission in AUD development and withdrawal, ALLO, acting as a positive allosteric modulator on GABAA receptors, may assist in restoring the balance between inhibition and excitatory neurotransmissions. ALLO, known for its anxiolytic, antidepressant, sedative, and anticonvulsant properties, experiences disrupted biosynthesis due to chronic alcohol consumption. Pregnenolone, its precursor, was shown to reduce craving and normalize autonomic arousal in AUD individuals, hinting at the potential contribution of ALLO and related neurosteroids to AUD development and recovery. However, clinical evidence regarding associations between ALLO, its GABAergic isomers, and precursors with AUD, including craving and treatment response to acamprosate, remains unknown. This project aims to explore the relationships between circulating neuroactive steroid levels and AUD, as well as AUD-related phenotypes, using existing AUD cohorts recruited for two NIAAA-funded clinical trials, including a discovery cohort (n = 297) and a validation cohort (n = 126). We will measure the serum levels of ALLO, its precursors, and its isomers by sensitive gas chromatography coupled with mass spectrometry. This neuroactive steroid panel will enable the evaluation of ALLO biosynthesis capacity and the excitation-inhibition balance on GABAA receptors. Specifically, we will investigate the association of serum ALLO and related neuroactive steroids with AUD by comparing these neuroactive steroid levels and their ratios between the discovery AUD cohort and age-and- sex-matched controls (Specific Aim 1). Focusing on the AUD cohort, we will examine the association of these neuroactive steroid levels and their ratios, with AUD-related phenotypes, including recent alcohol consumption, withdrawal symptoms, alcohol craving, sleep quality, and depression and anxiety levels, which were collected by widely used standardized questionnaires (Specific Aim 2). Finally, leveraging the follow-up clinical data and biospecimens collected, we will explore the associations of these neuroactive steroid levels and their ratios with treatment outcomes in individuals with AUD treated with acamprosate versus placebo (Specific Aim 3). This project will deepen the understanding of the effects of AUD and prolonged abstinence on the regulation of ALLO and related neuroactive steroids, thereby laying the groundwork for utilizing neuroactive steroids as biomarkers for treatment response and uncovering novel therapeutic mechanisms in AUD management.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT The objective of this study is to conduct a pragmatic, randomized clinical trial to evaluate the effectiveness and safety of oral midodrine in patients with sepsis associated hypotension. This patient-centered study approach is seeking alternatives to minimize the burden of intensive care unit (ICU) stay in these patients. Sepsis is one of the leading causes of death and disability worldwide. Cardiovascular compromise in sepsis manifests as hypotension due to arterial vasodilation. Hypotension often persists despite initial fluid resuscitation, prompting additional fluid boluses and subsequent administration of intravenous vasopressor agents. Although peripheral intravenous access could be used to initiate vasopressor treatment, the need for close blood pressure monitoring requires ICU admission and, for any but very short-term use, arterial and central venous catheter placement. Both excess intravenous fluid and arterial and venous catheterization may expose the patients to risks, unintended harms and discomfort. Moreover, a persistent need for intravenous vasopressor infusion prolongs ICU stay with associated burdens and cost. Midodrine is an oral vasopressor agent originally approved for use to treat orthostatic hypotension. Observational studies have suggested increasing off-label use of oral midodrine as a vasopressor-sparing agent in various groups of critically ill patients. Randomized controlled trials showed mixed results likely due to the heterogeneous general ICU patient population. Our preliminary data and the existing evidence point to clear equipoise and the need for conducting the proposed pragmatic single-blinded randomized clinical trial to evaluate both the safety and effectiveness of midodrine for sepsis associated hypotension. Our hypothesis is that administering oral midodrine to patients with septic shock who have received initial fluid resuscitation, appropriate antimicrobial treatment, and source control will decrease the duration of intravenous vasopressor treatment. If efficacious, midodrine can facilitate less invasive sepsis treatment and decrease the burdens and cost of ICU stay.

NIH Research Projects · FY 2025 · 2025-09

Project summary Age-associated B cells (ABCs) is a recently described B cell subset that accumulate during aging or development of systemic autoimmunity. They contribute to dysregulation of humoral immunity during aging and production of autoantibodies in autoimmune diseases. They are particularly responsive to TLR signaling and can be generated in both T cell dependent and T cell independent manner. Its metabolic and transcriptional regulation remain poorly understood. The MiTF/TFE transcription family member TFEB is a key modulator of lysosomal biogenesis, autophagy and lipid metabolism. We present preliminary evidence suggesting a potential contribution of TFEB transcription factor to the metabolic fitness of ABCs. TFEB overactivation leads to abnormal mitophagy and impaired mitochondrial metabolism in B cells, and dampened humoral immunity against T cell independent antigens. ABCs from aged mice, aged human donors and patients with systemic lupus erythematosus (SLE) exhibit elevated TFEB expression and its associated transcriptional signature. Our central hypothesis is that TFEB modulates systemic autoimmunity partly by controlling the mitochondrial metabolism and function of ABCs. Specifically, increased TFEB activity will reduce mitochondrial metabolism of ABCs, reduce autoantibody production and ameliorate SLE pathology, whereas reduced TFEB activity will have the opposite effects. In Aim 1, we will perform metabolic assays, transcriptomics and metabolomics to define the functions of TFEB in ABC generation, mitochondrial metabolism and function using novel genetic gain-of-function, loss-of-function, and reporter models. In Aim 2, we will test how TFEB may contribute to SLE pathogenesis using a TLR7 ligand induced lupus model on TFEB gain-of-function and loss-of-function mouse models. Our research will establish the first metabolic metabolism controlling ABCs.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT The prevalence of Type 2 diabetes (T2D) is an expanding global issue that continues to impact several developing nations. The onset of pancreatic β-cell functional failure is a central feature to the pathophysiology of T2D and is mediated in part by diabetogenic stressors including chronically elevated free fatty acids (lipotoxicity), low-grade systemic inflammation, and proteotoxicity. Due to their high protein synthetic burden, β- cells employ autophagy and endo/lysosomal pathways to reduce protein aggregates and protect β-cell mass and function. Recently, two endomembrane systems – small extracellular vesicles (sEVs; 30-150 nm sized nanoparticles) and autophagy have been shown to work in tandem to facilitate cellular adaptation and intercellular communication. Under diabetogenic stress, β-cell defects in autophagy mediated by auto/lysosomal dysfunction have been shown to occur, while conversely our group and others have noted enhanced sEV secretion in these conditions. However, what remains elusive is our understanding of the molecular mechanisms underlying the dis-balance between autophagy and sEV secretory pathways and potential therapeutic strategies to re-balance both in order to improve functional β-cell mass. Thus, our central hypothesis is that diabetogenic stress-mediated β-cell lysosomal dysfunction occurs in part, through activation of a novel secretory autophagy pathway to enhance β-cell sEV secretion. In Specific Aim 1, we will use in vitro and ex vivo, mouse and human diabetic models to investigate the molecular mechanisms associated with diabetogenic stress-mediated enhanced β-cell sEV secretion through activation of a novel sEV-based secretory autophagy pathway. We will use state-of-the-art EV based tools and technologies to visualize and quantify events that will determine activation of secretory autophagy. Furthermore, we will use sophisticated cell biology techniques to delineate the mechanism of preferential β-cell sEV release mediated by diabetogenic stress-induced amphisome de- acidification events. Our preliminary data reveal significant improvements in β-cell function and a reduction in β- cell sEV concentration under diabetogenic stress with pharmacological agents to either repress sEV biogenesis/release or enhance autophagy. Thus, in Specific Aim 2, we will explore in vivo and in vitro pharmacological, nutritional, and genetic therapeutic approaches to rebalance sEV secretory:autophagy pathway flux under diabetogenic conditions with the goal of improving functional β-cell mass. Using these approaches, we will first test the impact of a) inhibition of sEV biogenesis/release; b) enhancing autophagy; c) using a combinatorial approach to determine alterations in sEV release, autophagy induction, and β-cell function. Taken together, completion of the proposed application will provide novel insight into the molecular mechanisms that regulate sEV biogenesis and autophagy crosstalk during β-cell functional failure in T2D. Moreover, these studies will uncover new potential therapeutic approaches to restore the balance between both pathways with the goal to preserve and/or restore functional β-cell mass.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY This research proposal aims to evaluate vagal nerve stimulation (VNS) as a novel treatment for Tobacco Use Disorder (TUD), targeting improved smoking cessation rates. Cigarette smoking remains a major public health issue, leading to over 400,000 premature deaths annually in the U.S. and contributing to diseases such as lung cancer, chronic obstructive pulmonary disease (COPD), and heart disease. Despite widespread awareness of the risks and a strong desire to quit, many smokers are unsuccessful in their attempts, with only a small percentage achieving long-term abstinence. Current FDA-approved treatments—nicotine replacement therapy (NRT), varenicline, bupropion, and transcranial magnetic stimulation (TMS)—offer limited success in terms of efficacy and accessibility. There is a clear need for more effective and accessible treatments to support smoking cessation in TUD. This study's innovation lies in exploring VNS as a neuromodulation technique for treating TUD. Preclinical studies suggest that the vagal nerve is involved in addiction pathways, making it a promising target for smoking cessation interventions. This study will be the first to test VNS as a treatment for TUD using a non-invasive, handheld, and portable device, addressing key barriers to existing therapies like TMS, which requires specialized centers, and surgically implanted VNS devices. By using a pragmatic, decentralized clinical trial (DCT) approach, the study will combine VNS with existing pharmacotherapies, such as NRT and varenicline, to enhance smoking abstinence rates. The proposed phase-II randomized, double- blind, parallel-group clinical trial will assess the efficacy of NRT plus active VNS versus NRT plus sham VNS in smokers. Participants will receive 12 weeks of nicotine patch treatment combined with VNS, with an additional 12 weeks of varenicline and VNS for those who do not achieve abstinence. The trial will utilize remote carbon monoxide (CO) monitoring and telehealth visits to create a fully decentralized clinical trial environment, allowing for a patient-centered approach. Recruitment will occur through the Mayo Clinic’s Nicotine Dependence Center, leveraging our extensive experience in smoking cessation research. The primary endpoint of the study is biochemically confirmed smoking abstinence at 12 weeks, with secondary outcomes including prolonged abstinence at 24 and 26 weeks. This trial aims to provide preliminary evidence for the efficacy of combining VNS with NRT and varenicline, potentially leading to large-scale trials and FDA approval of VNS as a viable treatment for TUD. If successful, this research could add a novel, effective, and accessible therapy to the limited treatment options currently available for smokers attempting to quit.

NIH Research Projects · FY 2025 · 2025-09

To achieve the goals articulated in the NIH Advancing Science for the Health of Women strategic plan and advance our understanding of the antecedents of chronic disease in women, innovative strategies that leverage longitudinal data in large diverse populations are required. Cardiovascular disease (CVD) is the single largest killer of women. Clinically used risk scores used to guide treatment and prevention strategies to delay or prevent CVD have variable performance across demographic characteristics. Tools that provide more precise estimates of individual risk in women could significantly reduce the risk of CVD. Numerous clinical and imaging biomarkers are routinely measured serially across the lifespan but are typically confined to screening, diagnosing, or monitoring single diseases. Breast arterial calcification (BAC) is a mammographic biomarker that is associated with CVD. This application focuses on detecting and quantifying BAC and studying the utility of BAC for predicting CVD in women. Screening rates of mammography are high throughout the nation in every demographic group and thus represent a unique opportunity to directly interact with women who might otherwise underutilize preventive health care services. In comparison with 2D mammogram, over 80% of the imaging sites now also acquire digital breast tomosynthesis (3D) images. Our overarching hypothesis is that automated quantification of BAC from 2D and 3D mammograms will allow opportunistic prognostication of CVD risk. Therefore, we propose to study 125,519 women from two geographically and socioeconomically diverse cohorts from the University of Mississippi Medical Center and the Upper Midwest region via the Rochester Epidemiology Project. Our access to currently available clinical and imaging data uniquely positions us to 1) extend and validate the AI algorithm to detect and quantify the extent of BAC on both 2D and 3D breast mammograms; 2) identify clinical, demographic, and socioeconomic factors associated with BAC prevalence and progression; and 3) assess the predictive value of BAC as an independent biomarker of cardiovascular risk in aging women. The infrastructure in place at each site to extract and process images will enable the assessment of BAC in over 500,000 index and serial mammograms. Finally, we will follow these cohorts for over a decade for the development of CVD risk factors and events to determine the impact of BAC prevalence and progression across the lifespan on subsequent disease. If our proposed studies show that BAC is a significant early predictor of CVD in women, BAC measures could become an important way to refer women to preventive CVD therapies at the time of mammography. As such, mammogram screenings could become a way to not only address risk of breast cancer but also become part of primary CVD prevention.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract: Exercise intolerance and exertional dyspnea are hallmarks of heart failure with reduced ejection fraction (HFrEF). HFrEF patients have impaired locomotor muscle blood flow that directly contributes to submaximal exercise intolerance (and thereby limiting activities of daily living). Patients with HFrEF also exhibit exaggerated inspiratory muscle oxygen cost of breathing – a candidate mechanism contributing to exertional dyspnea in HFrEF. American College of Cardiology/American Heart Association guidelines recommend cardiac rehabilitation (CR) for patients with HFrEF (Class I recommendation). A core component of CR is exercise training. However, exercise training has a minimal impact on locomotor muscle blood flow and exertional dyspnea during submaximal exercise in HFrEF. Inspiratory muscle dysfunction is highly prevalent in HFrEF. In this context, inspiratory muscle training (IMT) is a clinically relevant intervention that targets the inspiratory muscles. IMT improves inspiratory muscle function, exercise tolerance, and quality of life in HFrEF. Importantly, IMT combined with exercise training leads to greater improvements in exercise tolerance and exertional symptoms than exercise training alone in patients with HFrEF. Our scientific premise is that the addition of IMT to CR increases locomotor muscle oxygen delivery and reduces inspiratory muscle oxygen cost of breathing during exercise contributing to the improvements in exercise tolerance and exertional symptoms with IMT combined with CR in HFrEF patients. The Specific Aims that will be explored in this proposal include: 1) To test if IMT combined with CR improves locomotor muscle blood flow during exercise compared to CR alone in patients with HFrEF, and 2) To test if IMT combined with CR reduces the exaggerated inspiratory muscle oxygen cost of breathing and improves ventilatory function to a greater extent than CR alone in patients with HFrEF. Both Aims are framed with testable hypotheses and clearly associated with the experimental protocol and statistical analysis plan. Our integrative, highly collaborative research team has the intellectual and technical expertise, established infrastructure, and clearly demonstrate high feasibility in performing all facets of these studies to address and interpret the Aims we have proposed. Our proposal addresses an important problem by focusing on ideas that are a significant departure from current paradigms on physiologic adaptations to exercise training in HFrEF patients. Our preliminary data and review of the rigor of prior research supporting our Aims provide strong justification for our innovative experimental design and gold standard techniques to reveal the physiologic mechanisms contributing to the improved exercise tolerance and exertional symptomology with IMT combined with CR in HFrEF. Finally, we have aligned our scientific premise, aims, and associated hypotheses with the NHLBI Research Priorities and the current NIH review criteria that emphasizes significance, impact, and innovation for R01 applications.

NIH Research Projects · FY 2025 · 2025-09

Augmented computing power, storage capability, and predictive analytics have accelerated adoption and deployment of Artificial Intelligence (AI) in health care delivery including for pediatric asthma care, the most common pediatric chronic disease with significant health disparities by socioeconomic status (SES), rurality, and demographics. Major concerns exist regarding the possibility that AI models perform systematically worse on disadvantaged populations, and the American Academy of Allergy, Asthma & Immunology, along with the Joint Commission, the National Academy of Medicine, and other non-government agencies, have expressed concerns that large-scale application of such unfair AI models may have substantial impact on health disparities. Ensuring fairness in AI is a crucial step toward improved asthma care for all children. Despite these concerns, the science of AI fairness in health care settings is still in its infancy. In our existing work, we applied current approaches of measuring and mitigating differential AI performance to an AI model for pediatric asthma care. We found the current measurement approaches to be inadequate, and the mitigation strategies to be unsuccessful. Three key limitations of the current approach for measuring and mitigating differential AI model performance in health care are: 1) statistical power and uncertainty measurement are not considered when assessing AI fairness metrics; 2) a lack of understanding of the role of EHR quality by sensitive attributes (e.g., SES) on differential AI performance; and 3) a lack of a suitable mitigation framework and strategies to reduce disparities by addressing the underlying sources of the disparities (EHR quality). Our study aims to address each of these knowledge gaps. Aim 1 is to develop a framework and tool for measuring differential AI performance using a statistical approach. We take our existing AI models for pediatric asthma exacerbation and asthma prognosis, and test for statistically significant differences by sensitive attributes. In addition to rurality and demographics, we will assess the role of SES in differential AI performance as the primary sensitive attribute by using the individual HOUsing-based SES (HOUSES) index, a validated, standardized, objective, and nationwide individual-level SES measure. We will do so via in-progress analytic and computational extensions of biostatistical techniques to estimate uncertainty and statistical power around AI fairness metrics. Aim 2 is to assess the role of EHR data quality by sensitive attributes (e.g., SES) on AI model performance. Using explanatory regression modeling (primary approach) and direct standardization (secondary approach), we will measure the strength of association between SES and AI model errors and see if this association is mediated by metrics of EHR quality. Aim 3 is to determine whether addressing EHR quality can achieve AI model fairness. This endeavor will enable AI to be used in fair and responsible ways to deliver optimal health care and achieve improved health for all children with asthma.

NIH Research Projects · FY 2025 · 2025-09

Our Networking Core (NC) will function synergistically with the Professional Development (PDC) and Training Cores (TC) to foster community and collaboration among trainees. Our leadership team has worked to ensure that networking mechanisms, including those led by trainees, underlie and are infused into the curricula of both Cores. Our NC will create infrastructure to support a bridge that joins trainees of this Program with others in the KUH research space, including the Collaborating for the Advancement of Interdisciplinary Research in Benign Urology (CAIRIBU) Community, a multi-institutional, multi-center collaborative of Urology O’Brien Centers, P20 Centers, and K12 Career Development Programs directed by Dr. Penniston (GENERATOR NC co-director) (Table N1). Our GENERATOR Program is unique since both institutions have currently-funded CAIRIBU Centers and Programs. Collectively, these Centers and Programs afford this NC a unique ability to directly leverage existing outreach programs and promote new interactions and research opportunities among GENERATOR Program urology trainees. To serve all of our KUH trainees we will also build relationships with leaders of the Cooperative Centers of Excellence in Hematology (CCEH)1 and its central coordinating center (University Utah) and the National Coordinating Center for the George M. O'Brien Kidney National Resource Centers (University of Alabama at Birmingham). To accomplish these goals our GENERATOR Networking Core Specific Aims are: AIM 1. Complement the training of 21st century collaboration and communication skills with practicum environments. Our primary objective is to develop opportunities for trainees to operationalize, practice, and sharpen the collaboration and communication skills learned in our Program. AIM 2. Encourage and foster a virtual trainee-led “community of practice” and other forums that afford trainees an unsupervised forum to relate to one another and interact. Our primary objective is to provide a forum to enhance trainees’ relatedness and communication. AIM 3. Share metrics of successes, best practices, and opportunities for improvement. Our primary objective is to comprehensively evaluate all elements and outcomes of our initiatives. Our Networking Core (NC) will function synergistically with the Professional Development (PDC) and Training Cores (TC) to foster community and collaboration among trainees. Our leadership team has worked to ensure that networking mechanisms, including those led by trainees, underlie and are infused into the curricula of both Cores. Our NC will create infrastructure to support a bridge that joins trainees of this Program with others in the KUH research space, including the Collaborating for the Advancement of Interdisciplinary Research in Benign Urology (CAIRIBU) Community, a multi-institutional, multi-center collaborative of Urology O’Brien Centers, P20 Centers, and K12 Career Development Programs directed by Dr. Penniston (GENERATOR NC co-director) (Table N1). Our GENERATOR Program is unique since both institutions have currently-funded CAIRIBU Centers and Programs. Collectively, these Centers and Programs afford this NC a unique ability to directly leverage existing outreach programs and promote new interactions and research opportunities among GENERATOR Program urology trainees. To serve all of our KUH trainees we will also build relationships with leaders of the Cooperative Centers of Excellence in Hematology (CCEH)1 and its central coordinating center (University Utah) and the National Coordinating Center for the George M. O'Brien Kidney National Resource Centers (University of Alabama at Birmingham). AIM 1. Complement the training of 21st century collaboration and communication skills with practicum environments. Our primary objective is to develop opportunities for trainees to operationalize, practice, and sharpen the collaboration and communication skills learned in our Program. We will create small, structured, and supervised transdisciplinary groups focused on specific research questions, skills-building topics, journal clubs, and collaborative grant-writing groups. Most will be virtual to allow cross-institutional participation. Faculty mentors in our Program will be assigned responsibilities to oversee and lead these activities. Existing CAIRIBU resources and activities will be leveraged to develop resources for all GENEATOR KUH trainees. AIM 2. Encourage and foster a virtual trainee-led “community of practice” and other forums that afford trainees an unsupervised forum to relate to one another and interact. Our primary objective is to provide a forum to enhance trainees’ relatedness and communication. We will encourage trainees to recognize their shared experience, build relationships and networks, and engage in joint activities. We will seed the concept for this “community of practice,” share evidence for its benefits, provide logistical support and coordination, and address needs that trainees identify. Structured, peer-led activities and informal collaboration among trainees can meaningfully impact professional development and the transition to research independence. Such opportunities for trainees to engage in self-directed initiatives and collaborative groups promote a sense of engagement, accountability, and momentum in their learning. Accordingly, we will actively encourage, support, and coordinate trainee-led initiatives that reinforce these goals.. AIM 3. Share metrics of successes, best practices, and opportunities for improvement. Our primary objective is to comprehensively evaluate all elements and outcomes of our initiatives. We will collect data to address high priority questions of the KUH communities. We will evaluate the structured practicum opportunities we create for trainees to practice and sharpen skills (Aim 1) and peer-initiated and led activities to develop trainees’ motivation for learning and success (Aim 2). We will disseminate our findings in manuscripts and presentations.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT This application, submitted by a multi-disciplinary research team, responds to RFA-DK-25-022: Precision Medicine for Type 1 Diabetic Nephropathy (U01 Clinical Trial Not Allowed). The goal of the RFA is to expand the Kidney Precision Medicine Project (KPMP) by adding new research sites to recruit individuals with Type 1 Diabetes (T1D) at various stages of or high risk for Diabetic Nephropathy (DN) for a longitudinal study involving research kidney biopsies at enrollment. Our institution is well-positioned to enroll a diverse T1D population, supported by robust electronic databases for participant recruitment and phenotype determination. We have extensive experience with clinical trials in the T1D population, enhancing our ability to recruit and retain participants. Resources include a T1D registry and the Expanded Rochester Epidemiology Project (E-REP), covering 27 counties in Minnesota and Wisconsin, linking medical records for over 60 years. This includes populations from rural regions with lower socioeconomic status, often underrepresented in clinical research We have integrated the HOUsing-based SocioEconomic Status (HOUSES) index and Rural-Urban Commuting Area (RUCA) classifications into a Technology-Enabled Subject Recruitment System (TESRS) to enhance recruitment efficiency from rural and under-resourced populations. In addition, we have access to Institutional cohorts performing genetic testing with archived samples and laboratory measurements including renal function. Support from ethicists experienced in biorepository consent and genomic research result return, along with patient advocacy partnerships, will help implement KPMP protocols for kidney tissue biopsy procurement. The development of surrogate markers for early and progressive kidney injury, including novel image-based biomarkers and cardiovascular (CV) events proposed by us aims to clarify disease pathways and identify high- risk T1D patients for DN and CV morbidity and mortality. Results of this project may facilitate drug discovery and has potential to improve outcomes of people with T1D DN.

NIH Research Projects · FY 2025 · 2025-09

The GENERATOR (“Next Generation of KUH Researchers”) is a research and career building program focused on predoctoral and postdoctoral scholars in KUH research laboratories. The training program will complement and fully integrate resources at the Mayo Clinic of Rochester (MCR) and the University of Wisconsin-Madison (UWM) as well as resources available through the U24 Interactions Core known as “Collaborating for the Advancement of Interdisciplinary Research in Benign Urology (CAIRIBU; housed at UWM). GENERATOR will leverage expertise from CAIRIBU to develop analogous career development activities for our “K” an “H” trainees. The primary goal or GENERATOR is to create a complete training pipeline and community of KUH researchers, spanning high school, undergraduate, post-baccalaureate, DVM and MDs in training, graduate, postdoctoral, and early career faculty levels. The five Specific Aims are: 1) Construct a pipeline that integrates investigators at multiple levels spanning high school through early career faculty into the KUH research space; 2) Recruit, select and maintain a trainee pool in KUH foci through continued evaluation; 3) Recruit, select and maintain a faculty pool in KUH foci through continued evaluation; 4) Develop and implement an engaging curriculum to provide fundamental training in KUH diseases, responsible conduct in research, grant writing, and professional development; 5) Integrate, organize, and offer existing resources (MCR, UWM) in a personalized training plan. We are committed to building, maintaining, and nourishing the KUH training pipeline and improving communication, collaboration, and teaching across this pipeline for upcoming generators of students, fellows, and early career faculty.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Beginning in 2016 and expanding in 2021, the WHO Classification of Tumors of the Central Nervous System utilizes acquired tumor alterations to classify and grade adult diffuse glioma. Genomewide association studies (GWAS) have identified 30 total germline variants across Europeans and East Asians associated with risk of developing adult diffuse glioma and some with development of specific clinically relevant molecular subtypes. Interestingly, some of these germline variants reside in or near genes that also have acquired tumor alterations in brain tumors. While a GWAS by the three primary 2016 WHO clinical subtypes were conducted, clinical characteristics and outcome heterogeneity exist within each subtype. Furthermore, the updated 2021 WHO criteria now utilize acquired tumor alterations for determining tumor grade. Thus, a current gap is to evaluate the genetic predisposition of developing a brain tumor with these clinically relevant acquired alterations or a tumor with a high mutation burden. Additionally, the functional relevance of most of the 30 germline variants remains unknown, which limits knowledge regarding the biology of glioma and critical information to develop preclinical models. Thus, we will use deconvolution of bulk RNA expression data, single-cell sequencing data, and modern epigenetic data to evaluate all known and newly identified germline variants. We will also determine the clinical utility of germline variants via polygenic risk models. While the prevalence of adult diffuse glioma is too low to screen the general population, we propose that a screening test can be used on targeted populations where differential diagnosis is challenging. Second, we propose that germline variants can predict whether a brain tumor patient has a particular acquired tumor alteration, e.g., IDH mutation, prior to surgery. Third, our preliminary data shows that polygenic risk scores are associated with overall survival. We will utilize both in-house and publicly available data on patient-matched germline and tumor specimens from adult diffuse glioma patients to expand our previous work and answer these additional essential questions. The project has the following three aims: Aim 1: Identify and validate novel germline variants associated with development of adult diffuse glioma with clinically relevant acquired tumor alterations. Aim 2: Using in-silico approaches, fine- map and evaluate the functional impact of known and newly identified adult diffuse glioma germline variants, accounting for specificity of cell type. Aim 3: Develop and validate the clinical utility of a glioma polygenic risk score for differential diagnosis of indeterminate brain lesions and to predict prognosis, and a polygenic tumor subtype model for predicting acquired tumor alterations. Sex and genetic ancestry will be carefully considered. Overall, understanding the interaction between germline and acquired genetics is crucial for development of more accurate diagnosis, targeting of modern glioma therapeutics that leverage specific tumor alterations, and improving patient outcomes.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract While heart transplantation is the gold standard to treat patients with end-stage heart failure, there is a shortage of donor hearts for recipients. Unfortunately, a minority of donor hearts on offer are accepted for transplant. This can be for multiple reasons, but the risk of primary graft dysfunction (PGD) is a major factor in turning down donor hearts. The risk of PGD is increased when cold static preservation time is >4 hours although machine perfusion technique can prolong this time by several hours. Even with these advances, a finite and significant geographical limitation is placed on organ matching. Expanding our molecular understanding of cardiac preservation is needed to position the organ preservation field for leaps in therapeutic advancements. Brd4 is a member of the bromodomain and extraterminal domain (BET) family of epigenetic readers. It plays an important role in many cardiac diseases such as cardiac hypertrophy and coronary atherosclerosis. Brd4 interacts with numerous factors that regulate transcription, histone modification, chromatin accessibility and architecture. We show that human failing hearts and donor hearts with prolonged preservation times have preferential short Brd4 isoform (Brd4-S) expression. We show that in-vivo knockdown of Brd4-S in cardiomyocytes greatly improves ex-vivo donor heart function with prolonged preservation. We also show that cold cardiac perfusion with histidine-tryptophan-ketoglutarate improves ex-vivo donor heart function and is associated with a significantly reduced Brd4-S expression. We hypothesize that cold preservation of donor hearts induces a switch towards Brd4-S expression leading to genome architectural restructuring with epigenetic changes that promote cardiac injury. In Aim1, we will determine if increased osmolality, cold perfusion and expression of splicing mediators contribute to Brd4-S expression during cardiac preservation. We used ex-vivo perfusion and heterotopic transplant models to test these hypotheses. In Aim 2, we will define the chromatin configuration induced by Brd4 isoforms during cold static cardiac preservation. We will use mass spectrometry to identify Brd4-S binding partners in donor heart preservation. To characterize the genomic landscape, we will examine the genome occupancy of Brd4-S and its cofactor, as well as histone modification, chromatin accessibility and looping in preserved donor hearts with Brd4 isoform knockdown. To determine the specific effects of cold perfusion on genomic organization, we will perform similar studies in cold HTK-perfused hearts. In Aim 3, we will determine if inhibition of Brd4 improves the preservation quality of pig and human hearts in a cold cardiac perfusion model. The proposed work will define Brd4-S's role in modulating chromatin configuration during cardiac preservation and represents a preclinical organ preservation study of combined Brd4 inhibition and cold HTK perfusion. This is expected to increase donor heart utilization and expand the donor pool with broad implications for other solid organ transplants and ischemic pathologies.

NIH Research Projects · FY 2025 · 2025-08

Modified Project Summary/Abstract Section This project investigates the association between oral microbiota and i) biomarkers of ADRD pathophysiology and ii) mid-life cognitive function. Substantial evidence links periodontal disease—a clinical condition characterized by an imbalanced or “dysbiotic” oral microbial ecology—with cognitive functioning and Alzheimer’s Disease and Related Dementias (ADRD). It is biologically plausible that alterations in the oral microbiota itself—which underlies periodontal disease—could contribute to ADRD pathology through both indirect and direct mechanisms. Indirect mechanisms include inflammation, hemodynamic alterations, and/or impaired glucose regulation, all of which are hypothesized to be impacted by the oral microbiome and to predict poor cognitive outcomes. There is also evidence that oral pathogens can directly impact the risk for ADRD by infecting the brain and inducing β-amyloid production and neurotoxic effects on tau proteins. This project addresses limitations of prior research by studying a large, diverse, representative cohort with direct measures of the oral microbiota and by studying biomarkers of ADRD pathology and objectively assessed cognitive functioning in the clinically normal range prior to ADRD onset. The proposed project efficiently leverages new data from the High School & Beyond (HS&B:80) cohort study. HS&B:80 has followed a nationally representative sample of ~25,500 Americans from high school in 1980 through age ~60 in 2021. Participants have been recontacted several times, with high rates of participation. In 2021, HS&B:80 gathered cognitive functioning, anthropometric, and biomarker measures including i) saliva-based microbiota assessments (n=~6,220) and ii) blood-based markers of ADRD pathophysiology (n=~4,220 who also have oral microbiota assessments). Markers of pathophysiological features of ADRD include Aβ42 and Aβ40, phosphorylated tau at threonine 181, neurofilament light chain, and glial fibrillary acidic protein. The project has two primary aims: (1) Investigate the association between the salivary microbiota composition and midlife cognitive functioning. (2) Investigate the association between the salivary microbiota composition and biomarkers of ADRD pathophysiology. Hypotheses: Oral microbiota signatures will be associated with cognitive functioning and biomarkers of ADRD pathology. Specifically, we posit a priori that elevated ratios of Treponema-to-Corynebacterium genera will be associated with i) reduced cognitive functioning and ii) ADRD biomarker concentrations that reflect amyloid and tau pathophysiology, axonal damage/neurodegeneration, and/or inflammation/astrocytosis. We will also consider whether early life education and family socioeconomic background account for observed associations in Aims 1 and 2. Hypothesis: Associations seen in Aims 1 and 2 will be partially attenuated after adjusting for early life education and family socioeconomic background.