University Of Minnesota

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract-Overall Coordinated and integrated care programs for individuals with Alzheimer's disease and Alzheimer's disease related dementias (AD/ADRD) and their family caregivers are growing. These models of care integrate disease care management, provide multidisciplinary care planning, and navigate healthcare and social service systems for persons living with dementia and their caregivers. However, knowledge of these services’ effectiveness is limited due to the lack of high-quality data at the state and sub-state levels, which hampers evaluation of policies that support integrated/coordinated dementia care services. Lack of data also inhibits opportunities to adapt and scale integrated/coordinated dementia care services within and across states. The State Alzheimer’s Research Support Center (StARS) will guide states through partnership, engagement, and 1-year pilot projects to build collaborations and create a shared data infrastructure to advance the accessibility, affordability, and effectiveness of dementia care services across the United States. Guided by the National Institute on Aging Health Disparities Research Framework, the specific aims are as follows: 1) to establish partnerships to identify existing coordinated/integrated dementia care services and available data sources within states; 2) to support up to 16 dementia care pilot projects; 3) to build a within- and across state data infrastructure to evaluate dementia care services and policies; and 4) to implement a multifaceted dissemination strategy to promote best practices in the identification, linking, and sharing of dementia care data. The State Alzheimer’s Research Support Center will create a national data infrastructure that enables states and all communities to better understand dementia care services’ effects on key care transitions, such as hospitalization readmissions, nursing home admission, home health care, as well as the health and well-being of people living with dementia and their caregivers. This infrastructure will also facilitate the examination of policy variations and changes on dementia care services within and across states. The partnership building, pilot project support, and data sharing infrastructure of StARS will ultimately result in greater dissemination of integrated/coordinated dementia care services that yield positive outcomes for people with dementia and care partners.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY Submitted in response to RFA-DA-24-063, we propose to develop novel, brain-penetrant, small molecule biased allosteric modulators (BAMs) of the neurotensin receptor 1 (NTSR1) to attenuate relapse to opioid seeking in individuals with opioid use disorder (OUD). NTSR1 is a G protein coupled receptor (GPCR) that is highly expressed in dopamine (DA)-rich brain regions and modulates brain DA signaling. NTSR1 ligands counter the effects of multiple classes of misused drugs. As a GPCR, NTSR1 signals via heterotrimeric G proteins and β- arrestin proteins. While NTSR1 has long been recognized as a promising target for the treatment of chemical addictions, development of balanced NTSR1 agonists that active both pathways is precluded by on-target side effects (i.e., hypothermia, hypotension). Our collaborative team developed a series of first-in-class β-arrestin BAMs of the NTSR1, which attenuate psychostimulant drug effects without the side effects characteristic of balanced NTSR1 activation. While NTSR1 is a well-established therapeutic target for stimulant use disorders, and its mechanism of action suggests utility that spans drug class, its validity as a target for OUD has not been rigorously established. The limited data available on NTSR1’s effect on opioid action is promising. Our data suggest that first generation β-arrestin NTSR1 BAMs act via a reward mechanism conserved across drug classes and attenuate both stimulant and opioid drug self-administration. In aim 1, we will validate NTSR1 as a drug target for the treatment of OUD with our optimized lead BAMWe will leverage a mouse model of relapse to intravenous (IV) opioid seeking, NTSR1 knockout (NTSR1-/-) mice, and our extensive knowledge of β-arrestin BAM pharmacology. Recently, we discovered that these compounds block NTSR1 signaling via some G proteins, but permit signaling via others. Because balanced NTSR1 agonists promote drug seeking, we hypothesize that this G protein permissiveness detracts from the therapeutic utility of these BAMs in OUD. In aim 2, we will discover next generation β-arrestin BAMs for NTSR1 with improved β-arrestin selectivity to test this hypothesis. We conducted comprehensive signaling characterization for a panel of ligands from our lead series. In addition, we have robust cell-based assays (and appropriate counter-screens) to reliably monitor NTSR1 activation of more than 14 transducers. Leveraging these assets, we will conduct a medicinal chemistry campaign to increase the potency and β-arrestin selectivity of BAM scaffold, with a flow scheme consisting of cell-based receptor signaling assays and early assessment of ADME, brain penetration, and central NTSR1 engagement. We have stringent criteria for second-generation leads. Compounds that match this profile will be advanced to efficacy testing in a model of relapse to IV remifentanil seeking in wild-type and NTSR1-/- mice. This multidisciplinary research plan capitalizes on the unique scientific and drug discovery expertise of our team and is a critical step towards our goal of developing therapeutics to facilitate recovery in individuals with OUD.

NIH Research Projects · FY 2025 · 2024-09

The US is currently grappling with an escalating drug overdose crisis, with overdose deaths soaring to a record high of 110,000 in 2022 alone. This crisis has particularly devastating effects on African American, American Indian/Alaska Native and other racial/ethnic minority populations. Harm reduction strategies, such as naloxone distribution, are pivotal components of the US Department of Health & Human Services Overdose Prevention Strategy. While many jurisdictions across the country are actively expanding the availability of evidence-based harm reduction programs, the complexity of the overdose crisis necessitates a multifaceted and localized approach that goes beyond individual interventions. The availability of over $50 billion in legal opioid settlements nationwide, alongside innovative harm reduction interventions, such as overdose prevention centers, drug checking programs, and overdose detection technologies, presents unprecedented opportunities to address the overdose crisis and its harms comprehensively. Our project aims to design tailored harm reduction strategies that effectively address the heterogeneous nature of overdose epidemics, demographics, and local infrastructures across six distinct jurisdictions. Building on our previous work optimizing naloxone distribution strategies in Rhode Island, Massachusetts, and New York City, we will collaborate closely with health departments and community partners to assess the effectiveness, cost- effectiveness, and health equity outcomes of different harm reduction strategies in these and three new jurisdictions (Minnesota, Missouri, and Nevada). Through adapting a mathematical model and developing locally tailored decision tools, we will identify portfolios of harm reduction strategies that minimize fatal overdoses, maximize health equity, and provide value for money. Central to our approach is stakeholder engagement and the implementation of evidence-based interventions tailored to local contexts. By engaging relevant stakeholders in the adaptation and customization of our model and decision tools, we seek to ensure their integration into decision-making processes and their sustainability beyond the duration of our project. Furthermore, guided by implementation science, we will assess the fit and scalability of the model and decision tools in supporting data- and equity-informed policy decisions. This project will create urgently needed, publicly available evidence and resources that can guide the allocation of opioid settlement funds and inform the adoption of sustainable and scalable harm reduction strategies. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions to the national opioid public health crisis. The NIH HEAL Initiative bolsters research across NIH to improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Biomedical breakthroughs over the past 30 years have dramatically improved treatment and prevention of HIV, hepatitis C virus (HCV), and opioid use disorder (OUD). Yet uptake of these HIV-related evidence-based interventions (EBI) remains distressingly low in populations that could benefit from them the most, notably people who inject drugs (PWID).Three structural limitations of current health delivery practices must be addressed to improve PWID access to HIV-related EBI: 1) the fragmented nature of the US healthcare system that forces PWID to seek care for infectious diseases and OUD from different entities; 2) the prohibitive administrative burdens of social programs such as Medicaid and Ryan White (complex eligibility requirements, burdensome paperwork); and 3) severe stigma toward PWID in healthcare settings that leads to care avoidance and poor retention in care. We propose an intervention that co-locates telehealth in syringe service programs (SSP) to improve uptake and retention of two key EBI, PrEP and MOUD. Because their practices are rooted in harm reduction principles and provide supplies that PWID routinely use, SSPs maintain strong client follow-up and offer many points of entry into care. Telehealth offers a more cost-effective, adaptable, and scalable model than in-person care, and recent service advances have generated a strong knowledge base for remote delivery of HIV-related care. Our intervention will further address administrative burdens by placing case managers on site to ease administrative burden, and by providing our telehealth providers with an evidence based stigma reduction training pioneered by our team. Our work will take place in four SSP sites across Minnesota which is currently experiencing an outbreak of HIV across racially and regionally diverse communities of PWID. Aim 1. Conduct formative research on determinants of HIV and addiction among racially and regionally diverse PWID to inform design of a co-located telehealth program. We will conduct in- depth interviews with N=25 PWID and N=10 advocates from PWID-facing organizations. We will also develop a tailored stigma reduction training for partnering providers using our proven experimental audit approach. Aim 2. Develop an integrative, low-barrier, low-stigma telehealth program for urban and rural PWID informed by Aim 1 findings and input from PWID and providers. Share back of Aim 1 findings will be conducted with 3 advisory boards (rural PWID, urban PWID, providers) to enlist their insights in the design of the stigma reduction training and key features of the telemedicine program. Aim 3. Pilot a telehealth intervention in racially and regionally diverse SSPs to assess feasibility, acceptability, and preliminary impact. Uptake and retention of PrEP and MOUD will be tracked for 6 months in 80 PWID across 4 SSPs (2 rural, 2 urban). Feasibility and acceptability will be informed by process indicators (enrollment, uptake, retention) and exit interviews. Preliminary impact will be assessed through a pre-post comparison of electronic medical records (EHR) from a state-wide data sharing platform.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Female genital mutilation/cutting (FGM/C) affects girls and women throughout the world, with 3 million girls cut each year. The highest rates of FGM/C are in East Africa. There are significant reproductive, sexual, and mental health consequences for girls and women impacted by FGM/C. Healthcare providers (HCPs) require training to meet these needs, yet globally there is a dearth of training programs. Effectiveness studies of FGM/C training are even rarer. Tanzania has a high FGM/C rate, yet we found in preliminary studies that healthcare students were unprepared to meet the need of these patients. The goal of the proposed study is to develop training materials specific to the needs of HCPs within Tanzania, deliver a pilot intervention/training program, and study the effectiveness of the program at Muhimbili University of Health and Allied Sciences (MUHAS). In order to meet that goal, it is critical that training materials are developed carefully, with attention to both cultural and ethical concerns facing healthcare providers within the country. There are three specific aims. In Aim 1 we will conduct a social ecological needs assessment of the healthcare needs of women in Tanzania who have experienced FGM/C. The results of Aim 1 will be used to adapt a promising training program to accomplish our second aim. In Aim 2 we will develop a FGM/C training tailored to the Tanzanian context. A core component of Aim 2 will be to use a train the trainer program. In Aim 3 we will evaluate the effectiveness of the training through a randomized, controlled, single blinded trial of the training (intervention) against a waitlist control arm (n=200 students per arm; 400 in total). In Aim 3 we will assess medical, nursing, and midwifery students’ improvements in knowledge, attitudes, and clinical skills. Development and testing of a tailored training course about FGM/C for students in health care, if effective, has high potential to be integrated into existing curriculum; and be widely adopted as a new standard of training for health professionals across both low prevalence and high prevalence FGM/C areas.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Stimulation of beneficial neuroplasticity is a crucial component in promoting recovery from neurological diseases or insults like spinal cord injury and stroke. Respiratory neuroplasticity, in the form of phrenic long-term facilitation (pLTF), can be induced non-pharmacologically via acute intermittent hypoxia (AIH). However, it is abolished in conditions of low circulating sex hormones or elevated systemic inflammation. At present, the interaction of sex hormones, CNS inflammation, and mechanisms of neuroplasticity are not well understood. The long-term objectives of this research are to understand the mechanism of microglial-mediated inflammation in cases of reduced circulating estrogen. Our central hypothesis is that reduced levels of circulating estrogen provoke a microglial-mediated increase in inflammatory cytokine gene and protein expression in the spinal cord, accompanied by altered microglial morphology, which inhibits plasticity. The hypothesis will be tested through two specific aims. 1: Define the inflammatory cytokine signature of the ventral cervical spinal cord of female rats across estradiol states. The approach for this aim will involve assessment of inflammatory-related gene and protein changes in the ventral cervical spinal cord of female rats using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and multi-color flow cytometry, respectively. Further, we will determine if cytokine changes are driven by spinal microglia by assessing isolated microglia from ventral cervical spinal cords of ovariectomized and naturally cycling female rats. 2: Determine the role of spinal cord microglia in modulating the expression of phrenic neuroplasticity in states of reduced estradiol. The approach for this aim will be to apply three dimensional Sholl analysis to cervical spinal cord segment images to allow quantification of altered microglial morphology across low and high states of circulating estradiol. Multi-color flow cytometry will be used to quantify microglial proliferation. Secondly, female rats will be treated with a CSF1R inhibitor to transiently eradicate CNS microglial populations. AIH-induced pLTF will be measured via in vivo phrenic neurophysiology. This proposed research will contribute to science by establishing the mechanistic role and molecular signature of spinal microglia in inhibiting plasticity in states of low estradiol. These contributions are expected to be significant in advancing neuroplasticity research because they will support the need to account for sexual dimorphisms when designing and implementing novel neurotherapeutic interventions. The proposed research is part of a fellowship training plan with extensive hands-on training and mentoring in technical and professional research skills necessary to become an independent clinician-researcher. Primary research activities and training will take place at the University of Minnesota Medical School with additional training at the University of Wisconsin – Madison. The University of Minnesota’s strong foundation in research and the support of the Rehabilitation Science graduate program will create an ideal environment for completing this training.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY RNA plays numerous roles in biology. In addition to its canonical role in the central dogma as an intermediary between DNA genomes and protein products of translation, a myriad of roles have been described for this molecule, including gene silencing (siRNA), regulation of translation (bacterial riboswitches), and the translation machinery itself (rRNA). Despite the importance of these roles in biology, tools for imaging RNA are not as robust as the suite of fluorescent proteins that exist for readout of protein machinery. This project seeks to address this need by developing new fluorescent aptamer tags for RNA visualization. This strategy has enjoyed considerable success, particularly in bacterial systems, but deployment in complex systems (eukaryotic, whole- organism) has been challenging. Recent work on RNA folding has shown that a noncanonical RNA structure (a G Quadruplex) may impair the use of aptamers as tags in these systems. This project seeks to select fluorescent aptamers that are not G-Quadruplexes. We will seek select non-G-quadruplex-forming aptamers against a range of azo dyes (Aim 1) and endogenous pigments (Aim 2) for robust live-cell imaging, and a long-range goal of whole-organism RNA imaging. We will use these tags to characterize Lin28-mRNA interactions, which is not possible with existing G-quadruplex-forming aptamers, because Lin28 is known to unfold G-quadruplexes in RNA. We will also develop benchmarking tags for our selected aptamers by selecting non-G- quadruplex-forming aptamers to known aptamer ligands, which we will use to compare our selected aptamers.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Heart failure with preserved ejection fraction (HFpEF) is one of the most urgent prevention and treatment challenges in public health today given its increasing prevalence, limited therapeutic options, and substantial burden on global health care systems. Exercise-based prevention and treatment strategies for HFpEF have historically focused on the promotion of aerobic exercise. However, given the coinciding epidemics of physical inactivity and obesity in the U.S., and the compromised aerobic capacity and exercise intolerance in patients with HFpEF, long-term adherence to aerobic exercise is generally poor and thus, the prevention and treatment of HFpEF remains a major public health challenge. Thus, aligned with NHLBI research priorities for HFpEF, research is critically needed to: (1) identify optimal strategies for improving adherence to exercise in the prevention and treatment of HFpEF; and (2) examine biological pathways linking exercise to HFpEF. In the proposed project, Dr. McDonough, an exercise science and public health researcher, will examine the isolated role of resistance exercise in the prevention and treatment of HFpEF with examination of potential biological intermediates. With the exceptional mentorship team he has assembled, comprised of prolific epidemiologists, physician scientists, and methodologists, and the resources available to him through the University of Minnesota, this Postdoctoral Career Transition Award to Promote Diversity will support Dr. McDonough in filling critical training gaps and conducting the research necessary to launch his career as an independent investigator utilizing cohort studies to inform his future clinical trials in the treatment and secondary prevention of HFpEF. To prepare him for this role, a multifaceted training plan including coursework, mentorship, and research is proposed in: (1) the analysis of longitudinal data from cohort studies using a variety of multivariable regression modeling approaches; (2) the pathophysiology and epidemiology of HFpEF and how to safely implement exercise training in patients with HFpEF; (3) advanced clinical trial methods and statistical analyses; and (4) advanced epidemiologic statistical modeling of pathways and mediators. The expertise Dr. McDonough develops through this training plan will be essential for conducting the proposed research. During the mentored K99 phase, he will leverage NHLBI cohort studies to elucidate the independent role of resistance exercise in HFpEF primary and secondary prevention (Aims 1-3). In the independent R00 phase, Dr. McDonough will conduct an independent feasibility clinical trial examining the effect of resistance exercise on liver fat and subsequent functional capacity, quality of life, and intervention adherence in HFpEF patients (Aim 4). Findings from this project will provide the requisite preliminary data to support an NIH R01 grant proposal examining novel exercise therapies for improving hospitalization, mortality, and physical activity outcomes in HFpEF patients, in line with NHLBI, AHA, and ACC research priorities. In turn, Dr. McDonough will be well-positioned for a successful transition to an independent research career at an R1 institution while helping to enhance diversity in the biomedical research enterprise.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract. The Myc gene family encodes three highly conserved transcription factors (N-, c-, L-), known to be oncogenic drivers in many cancers. Dysregulation of Myc proteins is estimated to drive 30% of all cancers, attributed to inappropriate amplified Myc expression and dysregulated behaviors. c-Myc is prominent in a wide variety of cancers, due to its broad expression patterns, where N-Myc and L-Myc are tissue specific and heavily associated in neuroblastoma and small cell lung carcinoma, respectively. Due to its prevalent cancer involvement, Myc family transcription factors are considered an attractive anti-cancer target, but their disordered nature makes them poor drug targets. Protein-protein interactions of Myc with other regulatory partners creates an opportunity for therapeutic intervention via indirect targeting, yet structural details of many Myc interactions remain unclear. The N- and c-Myc binding partner Aurora Kinase A (AurA), a serine-threonine kinase, is hypothesized to stabilize Myc by preventing proper ubiquitin-mediated degradation by the SCFFbxw7 ubiquitin ligase complex. In this pro- posal, fluorescence anisotropy and time-resolved fluorescence will be utilized to fully assess binding of AurA at varied phosphorylation states and lengths of c-Myc. To structurally characterize a c-Myc/AurA complex I will pursue extensive training in X-ray crystallography alongside my experience using continuous-wave electron par- amagnetic resonance (CW-EPR) spectroscopy to study the interface of this interaction, proposed within. My preliminary work has supported the formation of an AurA/c-Myc/Fbxw7 complex, of which I will structurally char- acterize using continued training in cryogenic-electron microscopy (cryo-EM). Further characterization of c-Myc stabilization and ubiquitination patterns by AurA using my developed in vitro ubiquitination assays will result in a defined mechanism of AurA induced stabilization of c-Myc. The high conservation of regulatory protein binding domains across Myc family transcription factors suggests a role of AurA stabilization of L-Myc, already identified for N- and c-Myc. The work and training I receive studying the c-Myc/AurA interaction will build foundations for my future independent research on defining an impact of AurA on L-Myc stabilization, a widely understudied Myc family member. I will also use in cellular work to characterize the physiological protein interactions involved in L- Myc regulation and dysregulation, including kinases and SCF ubiquitin ligase components, currently unidentified. The extensive X-ray crystallography and cryo-EM training proposed within will add a strong structural biology foundation to the independent research program I plan to develop at an R1 institution. The focus of my lab will be rooted in characterizing the regulation mechanisms of disordered transcription factors in cancer and disease. I believe the extensive structural biology and biophysical background from proposed training and extensive ex- perience will make me a competitive candidate for a future tenure-track faculty role.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY The past decade has witnessed exponential surges in social media use among U.S. adolescents. Meanwhile, as mental health problems increase for all ages, the adverse trend is particularly striking among adolescents. Despite vigorous discussion about the contribution of social media to worsening mental health in adolescents, little objective data exist about how adolescents use social media and how it affects mental health. Few studies have examined strategies adopted by adolescents to regulate and manage their social media use; few have used detailed and objective measures of social media use and content exposure; and almost no longitudinal studies exist incorporating the strategies, social media use, and mental health. To fill these critical gaps, we propose a 2-year longitudinal mixed-method study with a national sample of 400 adolescents aged 13-17 years. Phase 1 will collect preliminary data from 100 adolescents to establish measure feasibility and optimize research procedures (Aim 1a). In Phase 2, a full-scale longitudinal data collection will be completed with 300 adolescents. Phase 1 will focus on one 14-day-long epoch where we will (1) collect daily diary reports of social media management strategies (SMMS; targeted at access, duration, timing, risky content, and risky social relations) and the social sources and approaches/tools for SMMS implementation, (2) administer baseline and end-of-epoch surveys on mental health, and (3) conduct semi-structured interviews among a subsample. During Phase 1, we will also use our existing pilot data of 154 adolescents to establish algorithms for processing data from Phase 2 (Aim 1b). Phase 2 will involve a baseline survey and 8 14-day epochs (2 years X4 quarterly epochs/year). During each epoch, in addition to the SMMS daily diary reports and end-of-epoch mental health survey, we will also use the Screenomics approach to continuously capture screenshots of adolescents' phone screens every 5 seconds to measure multiple granular objective indicators of social media use. Screenomics was IRB-approved, implemented, and proven secure and feasible in our pilot study. Annual semi-structured interviews will also be conducted among a Phase 2 subsample. The Phase 2 design will enable the examination of daily reciprocities between the multiple SMMS domains and social media use indicators (Aim 2), and the bidirectional dynamics associating SMMS, social media use, and their reciprocities with mental health (Aim 3). This highly innovative project will provide urgently needed information regarding how adolescents manage their social media use on a daily basis, which strategies influence actual social media use, and how the strategies and social media use captured through objective data influence and are influenced by mental health. Findings will offer informative solutions for healthier and safer use of social media to educators, practitioners, parents, policy makers, technology companies, and adolescents themselves. This work will also directly inform interventions aimed at maximizing the benefits and minimizing the harms of social media on adolescents' mental health with effective strategies that are well received by adolescents.

NIH Research Projects · FY 2025 · 2024-09

Summary West Nile virus is a pathogen of global concern. Since its introduction to the US in 1999, it has caused thousands of deaths and tens of thousands of hospitalizations. West Nile virus is transmitted to humans by the bite of infected mosquitos; the virus mediates a febrile illness and can progress to invasive infection of the central nervous system causing severe encephalopathy. WNV neuroinvasive disease also causes cognitive and motor sequelae that can last for months or years after the virus is cleared. In order to successfully treat these symptoms, gaining a clearer understanding of the immune response to WNV and the factors that determine the initiation and severity of neuroinvasive disease is critical. There is incomplete understanding of what virus and host determinants control or mediate WNV neuroinvasion, how the immune response to WNV within the CNS is initiated and regulated, and how the acute response to WNV influences long-term inflammatory sequelae of WNV encephalitis. The goal of the proposed studies is to address these knowledge gaps by gaining a molecular understanding of WNV neuroinvasion, CNS immunity, and inflammatory sequelae. We will do this by deploying newly-developed unique mouse models, recombinant forms of WNV, and an array of single-cell and spatial transcriptomic and epigenomic methods. First, we will define the cells that initiate the immune response to WNV in the periphery, and the targets of the cytokines produced by this response that control WNV neuroinvasion. Second, we will define the signaling circuits by which WNV initiates and propagates inflammatory responses within the CNS. Third and finally, we will use a recombinant WNV strain expressing Cre recombinase to mark CNS cells that survive WNV infection and assess their contribution to persistent inflammatory sequelae that persist after WNV clearance. New understanding gained from the proposed studies will significantly advance our understanding of the determinants and consequences of WNV neuroinvasive disease, and will inform ongoing efforts to mitigate and ameliorate these consequences in human patients.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Catheter-based renal nerve ablation (CBRNA) decreases blood pressure (BP) and disrupts the chronic overactivity of sympathetic efferent renal nerves thought to underlie neurogenic hypertension (HTN), diminishing the risk for developing other cardiometabolic diseases. Studies have demonstrated that a contributor to the development of neurogenic HTN is sensory afferent renal nerves (ARNs) whose input to central autonomic regions modulates sympathetic outflow to other organs. Recent work from our lab has identified the close proximity of ARNs to glomeruli in the renal cortex, which is novel in comparison to the traditional view that ARNs primarily innervate the renal pelvis. The implication of ARNs in regulating BP and the identification of ARNs in the renal cortex indicates the need for the role of ARNs in kidney function and autonomic processes to be investigated. This proposal aims to address this gap in knowledge by 1) elucidating the central terminations and regional specificity of renal afferents, and 2) identifying how renal afferent input influences the activity of central autonomic regions that regulate blood pressure using a salt-induced model of hypertension. Aim 1 will use viral vector-based neurotracing to label ARNs from the renal cortex and renal pelvis of the kidney to identify their central terminations. I hypothesize that two distinct populations of ARNs innervate the renal cortex and pelvis respectively, and project to distinct primary central termination sites. Tracing will be visible throughout the neuroaxis including the renal vasculature, dorsal root ganglia, nodose ganglia, spinal cord, and brainstem. Aim 2 will be completed using the deoxycorticosterone acetate (DOCA)-salt mouse model, which induces HTN through sodium and water retention, in targeted recombination of active populations (TRAP2) mice where the fluorescent reporter protein tdTomato is expressed under the immediate-early gene c- Fos, a neuronal activation marker, via tamoxifen-inducible Cre. The expression of tdTomato will be quantified as a direct measure of c-Fos in brain regions known to be involved in BP regulation during early DOCA-salt HTN development and compared to immunolabeled c-Fos at a later timepoint. I hypothesize that DOCA will increase the expression of tdTomato and c-Fos in non-denervated mice, and that afferent renal denervation (ARDN) and an IL-1 receptor antagonist will both attenuate the DOCA-induced neuronal activity. The expected outcomes of this proposal are to have traced the terminals of ARNs from the kidney to central primary processing centers and identified changes in neuronal activation in central autonomic centers relevant for ARN processing. These results will expand the understanding of how ARNs are involved in salt-induced HTN and establish an anatomical foundation for future investigations into the physiological roles of ARNs in the autonomic control of BP regulation. Further, these findings may contribute to the potential mechanism by which ARDN attenuates increased BP in salt-induced HTN allowing the development of interventions that can more accurately target ARNs, both peripherally and centrally, increasing the therapeutic potential of ARDN as a treatment for hypertension.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT Chronic Low Back Pain (LBP) is a leading cause of disability worldwide and one of the most common reasons patients are prescribed opioids, despite their poor ability to improve function. LBP threatens our economic health due to high rates of health care utilization and disability, our security due to high rates of medical discharge from armed services, and our quality of life due to pain-related suffering, pain-associ- ated opioid misuse, depression, and anxiety. The primary driver in 40% of all LBP cases is estimated to be pain resulting from intervertebral disc (IVD) degeneration, referred to as discogenic LBP. Improved understanding of this degenerative process is needed to address this global problem. Epigenetic mecha- nisms, such as DNA methylation, dynamically regulate gene expression in response to local, systemic and environmental influences. However, very little is known about the role of DNA methylation in dis- cogenic LBP. This application will address this knowledge gap by determining the role of DNA methyla- tion in the pathological expression of genes involved in IVD degeneration and discogenic LBP. Our central hypothesis is that widespread and persistent reprogramming of gene expression by DNA methylation drives IVD degeneration and can be targeted for the treatment of discogenic LBP. Since DNA methylation is potentially a mechanism underlying the chronicity of pain, elucidation of the role of DNA methylation in IVD degeneration is likely to provide a strong scientific framework for innovative new treatment approaches for discogenic LBP. In Aim 1 we will identify the DNA methylation and gene ex- pression landscape of IVD degeneration and discogenic LBP in humans. In Aim 2, we will identify the DNA methylation and gene expression landscape of IVD degeneration and discogenic LBP in animal models and delineate correlated DNA methylation and gene expression profiles. In Aim 3, we will explore the role of DNA methylation on IVD degeneration and discogenic LBP in animal models by manipulating methylation using non-pharmacological (home cage voluntary running wheels) and pharmacological in- terventions. These results will be significant because scientific evidence of a role for epigenetic reprogramming in IVD degeneration and discogenic LBP may contribute to a paradigm shift in understanding, prevention, and treatment of chronic LBP.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY More than 40% of patients with diabetic retinopathy (DR) are refractory to direct intraocular injection of anti- VEGF antibodies, the standard of care. The inability to effectively treat DR puts a heavy burden on the health care system and limits the quality of life for those affected. The need for novel, clinically relevant targets that can be exploited for new therapies is very clear. Diabetes is well-known to induce retinal mitochondrial dysfunction that leads to DNA damage and cytosolic release. Cytosolic DNA activates the cGAS-STING pathway, inducing inflammation. Although the cGAS-STING signaling axis is essential for sensing foreign DNA and thus contributing to innate immunity, aberrant activation of this pathway by self-DNA is a significant contributor to autoinflammatory and autoimmune diseases. Activation of cGAS-STING due to chronic diabetes-induced inflammation leads to activation of circulating monocytes, eventually giving rise to leukostasis. Leukostasis causes cell death and breakdown of the blood-retinal barrier, leading to retinal vascular leakage, a major cause of diabetic macular edema and leading cause of vision loss in diabetic patients. We have shown that 1) the cGAS-STING pathway is activated in ischemic retinopathy; 2) inhibition of STING attenuates monocyte activation and alleviates retinal leukostasis and angiogenesis; 3) STING knockout significantly reduces retinal leakage, number of leukocytes in the retina, and DR relevant inflammatory factors in an oxygen-induced retinopathy murine model; and 4) small molecule agonism of peroxisome proliferator-activated receptor alpha (PPARa) inhibits the cGAS-STING signaling axis and cytosolic mitochondrial DNA release. The objective of this exploratory R21 proposal is to assess two mechanistically differentiated approaches to inhibit STING with bifunctional small molecules in the context of DR. In Aim 1 we will assess small molecule induced STING degradation through proteolysis-targeting chimeras (PROTACs) as a strategy for STING inhibition in the context of DR. The PROTAC strategy is unexplored in the eye but especially well-suited for ocular conditions. In Aim 2 we will leverage our success in developing selective PPARa agonists as new therapeutic leads for DR to develop first-in-class bifunctional probes that simultaneously agonize PPARa (neuronal protection) and inhibit STING (retinal vascular homeostasis and inflammation) to explore the potential benefits of polypharmacology in retinal disease. The heterobifunctional modulators of these two targets are expected to provide a spatiotemporal benefit affecting multiple cell types and pathological events. This work is conceptually and technically innovative and is expected to provide the fundamental data needed to enable proof-of-concept for the PROTAC and multi-action compound strategies in ocular contexts (new approaches), a foundation for targeting STING in DR and related retinal diseases (new target), and new compounds poised for biological and translational initiatives, respectively. This project leverages a long-standing and productive collaboration leveraging expertise in medicinal chemistry and biochemistry to advance knowledge regarding STING biology and therapeutic utility in DR.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The goal of this project is to refine our gene therapy strategy for Xeroderma Pigmentosum-Cockayne Syndrome (XP-CS), with the ultimate aim of improving the neurological outcomes and life expectancy for this disorder. XP- CS is a rare, inherited DNA repair disorder that is characterized by an accelerated aging phenotype and prominent, severe neurodegenerative complications mirroring those seen in classic Cockayne syndrome (CS), including cognitive dysfunction, brain atrophy, neurodevelopmental delays, peripheral neuropathy, brain atrophy, and dementia. There are several genetic subtypes of XP-CS, including one associated with pathogenic variants in the gene ERCC5 (XPG). We have found that the Xpg-/- mouse accurately recapitulates key features of the XP- CS phenotype, including neurological features. We have also developed a first generation adeno-associated virus (AAV) vector capable of delivering ERCC5 that shows great promise but requires further development prior to translation into human clinical trials. Our preliminary studies using the first generation vector in the Xpg-/- mouse point to a clear path towards optimizing this approach, including capsid optimization, promoter optimization, delivery optimization, dose tuning, toxicity studies, and biomarker evaluations. We expect that our proposed studies will lead to a gene therapy strategy that overcomes many of the obstacles to translation into a Phase 1 human clinical trial. The knowledge obtained from these studies will be generalizable, and will accelerate the development of gene therapies for other genetic subtypes of XP-CS.

NIH Research Projects · FY 2024 · 2024-09

Kidney stones are a painful and common health problem, occurring in 8-12% of the population. Most kidney stones are composed of calcium oxalate (CaOx). Current therapies to prevent CaOx kidney stones have low efficacy, and recurrence rates are high. Pet dogs are an exceptional model of kidney stone disease. They naturally form CaOx kidney stones and share environmental and metabolic risk factors with humans. Using this natural model, we discovered a genetic variant in the uromodulin gene that has a profound effect on stone risk. Uromodulin (aka Tamm-Horsfall protein) is the most abundant protein secreted in urine and has long been linked to human kidney stone disease. The canine stone-risk variant is located within a sequence of uromodulin that is necessary for polymerization and excretion of small bioactive peptides. Our preliminary work demonstrated that both polymerization and peptides are disturbed in dogs with the variant, implicating these uromodulin features in stone pathogenesis. Our overarching objective is to identify the renal cellular and molecular mechanisms by which uromodulin polymerization status and peptides impact stone formation. The proposed study will test the hypothesis that defects in uromodulin polymerization status and peptides cause alterations in renal solute transporters/channels (Aim 1) and inflammatory cytokines involved in stone formation (Aim 2). The results of this work will identify disturbances in renal solute transport and inflammatory molecules that occur with abnormal uromodulin polymerization and peptide excretion. This will provide important insight into how uromodulin features affect its functions in the kidney. Uromodulin polymerization and peptide excretion can be modified with diet or drugs. Thus, the proposed research will lay the groundwork for future development of therapies to mitigate risk for CaOx kidney stone recurrence in humans.

NIH Research Projects · FY 2025 · 2024-09

MN PRIMED is a unique postbaccalaureate program for recent college graduates, offering a two-year intensive research experience combined with advanced graduate courses. Leveraging UMN's extensive research expertise in Diabetes, Endocrinology, and Metabolism (DEM), MN PRIMED aims to shape the future generation of researchers in DEM-related research. Participants are paired with two of the 20 experienced faculty mentors, known for their DEM-related research leadership and commitment to mentoring trainees. This sets the stage for independent research projects, academic growth, and publication opportunities. To achieve MN PRIME’s goal of preparing participants for doctoral programs with a focus on diabetes research, MN PRIMED doctoral readiness program will employ a multifaceted approach: 1. Providing trainees with exceptional research experiences to develop laboratory competence and multidisciplinary research skills regarding DEM. 2. Offering high-quality mentorship, including senior/associate mentors (mentor dyads) to support trainee and associate mentor development. 3. Assuring a path to graduate program entrance with application process guidance and guaranteed interviews. 4. Facilitating skill development, such as manuscript writing and effective communication in science, while also helping trainees navigate the complexities of graduate school. This administrative supplement will provide four additional postbaccalaureate positions in Year 2 to enhance peer-to-peer engagement among scholars and increase the number of learners pursuing careers in medical research focused DEM.

NIH Research Projects · FY 2024 · 2024-09

PROJECT ABSTRACT Opioid Use Disorder (OUD) is a complex medical condition characterized by compulsive and maladaptive use of opioid substances, including prescribed pharmaceuticals (such as oxycodone) and illegal agents (such as heroin). It causes physiological, psychological, and sociological effects, including tolerance, withdrawal symptoms, relapse, and impaired daily functioning. Managing opioid use disorder (OUD) involves addressing withdrawal symptoms and relapse risks during abstinence, which can vary throughout different stages of recovery. During the initial phase of opioid withdrawal, individuals often encounter the most severe and debilitating symptoms. These symptoms typically peak within the first few days of refraining from drug use and gradually subside with prolonged abstinence. Nevertheless, the most significant challenge during this critical stage of recovery is the sustained risk of relapse, even after the initial withdrawal symptoms have abated. This risk is further amplified by external cues that can trigger intense cravings and the compulsion to seek drugs. Hence, it is imperative to acquire a comprehensive comprehension of the neural mechanisms that underlie relapse during early and prolonged abstinence from opioid use. Rodent studies suggest that glutamatergic projections from the paraventricular nucleus of the thalamus (PVT) to the nucleus accumbens (NAc) are involved in the expression of negative affective states and relapse after abstinence. Both PVT and NAc are heterogeneous and complex brain regions with diverse sets of cell types, functional connections, unique subregions, and neurotransmitter systems. Opponent roles of anterior/posterior PVT subregions, and D1- and D2-medium spiny neuron activity, has been found for approach/appetitive and avoidance/aversive behaviors. This research project addresses the critical question of how the PVT interacts with the NAc to modulate withdrawal symptoms during early abstinence and prolonged abstinence on the vulnerability to relapse. This project is an essential step towards my goal of becoming an independent researcher. Through the K99/R00 grant, I'll have the opportunity to improve my understanding and abilities in advanced neuroscientific methodologies, which will lay the groundwork for my research program focused on addiction neuroscience. The knowledge and data I gather during the K99 phase will be a solid foundation for a successful transition to the R00 phase, which will help me expand my research in this field. Additionally, this work will reveal the complex PVT-NAc neural mechanisms governing relapsing behaviors, which could lead to innovative strategies for addiction treatment and prevention.

NIH Research Projects · FY 2026 · 2024-09

PROJECT ABSTRACT Opioid Use Disorder (OUD) is a complex medical condition characterized by compulsive and maladaptive use of opioid substances, including prescribed pharmaceuticals (such as oxycodone) and illegal agents (such as heroin). It causes physiological, psychological, and sociological effects, including tolerance, withdrawal symptoms, relapse, and impaired daily functioning. Managing opioid use disorder (OUD) involves addressing withdrawal symptoms and relapse risks during abstinence, which can vary throughout different stages of recovery. During the initial phase of opioid withdrawal, individuals often encounter the most severe and debilitating symptoms. These symptoms typically peak within the first few days of refraining from drug use and gradually subside with prolonged abstinence. Nevertheless, the most significant challenge during this critical stage of recovery is the sustained risk of relapse, even after the initial withdrawal symptoms have abated. This risk is further amplified by external cues that can trigger intense cravings and the compulsion to seek drugs. Hence, it is imperative to acquire a comprehensive comprehension of the neural mechanisms that underlie relapse during early and prolonged abstinence from opioid use. Rodent studies suggest that glutamatergic projections from the paraventricular nucleus of the thalamus (PVT) to the nucleus accumbens (NAc) are involved in the expression of negative affective states and relapse after abstinence. Both PVT and NAc are heterogeneous and complex brain regions with diverse sets of cell types, functional connections, unique subregions, and neurotransmitter systems. Opponent roles of anterior/posterior PVT subregions, and D1- and D2-medium spiny neuron activity, has been found for approach/appetitive and avoidance/aversive behaviors. This research project addresses the critical question of how the PVT interacts with the NAc to modulate withdrawal symptoms during early abstinence and prolonged abstinence on the vulnerability to relapse. This project is an essential step towards my goal of becoming an independent researcher. Through the K99/R00 grant, I'll have the opportunity to improve my understanding and abilities in advanced neuroscientific methodologies, which will lay the groundwork for my research program focused on addiction neuroscience. The knowledge and data I gather during the K99 phase will be a solid foundation for a successful transition to the R00 phase, which will help me expand my research in this field. Additionally, this work will reveal the complex PVT-NAc neural mechanisms governing relapsing behaviors, which could lead to innovative strategies for addiction treatment and prevention.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT Technology will be developed to enable future compact, mid-field (0.7 Tesla) magnetic resonance imaging (MRI) systems, for improving human health worldwide. MRI is an indispensable imaging tool that provides measurement capabilities unavailable with other modalities. Yet, due to its expense, large size, and demand on facility infrastructure, high quality MRI remains inaccessible to a large fraction of the world’s population, particularly in remote and resource-limited settings. The existence of portable, affordable, high-performing MRI technology will substantially expand its accessibility for both clinical care and neuroimaging research. Although low field (<0.1 Tesla) MRI scanners are now commercially available, to date they have not produced images of similar quality as those of mid- and high-field MRI scanners unless relying on intensive post-processing based on machine-learning and AI, which makes the reliability of these low field images uncertain at this time. As an alternative approach for increasing portability, and thus increasing access to mid-field MRI (0.1 - 1 Tesla), in this project we will further develop new technology called FREE (Frequency-modulated Rabi-Encoded Echoes) that has potential to eliminate one of the most expensive and massive hardware components of an MRI system; namely, the pulsed field gradients that are conventionally used to encode spatial information in MRI. Instead, the MR signals will be encoded by spatially varying radiofrequency (RF) fields, using specialized multi-channel RF coils and a novel frequency-swept pulse technique that performs spatial encoding using RF field gradients, even when the magnet produces a highly nonuniform field. Further, this project will build upon the previous innovations by this same team in a U01 grant that led to: 1) the capability to perform MRI with extreme magnetic field inhomogeneity (~2-3 orders of magnitude greater than what is commonly perceived to be necessary), 2) a unique compact high temperature superconducting (HTS) head-only magnet, and 3) a state-of-the-art multi-channel digital spectrometer for programming and controlling the MRI scanner. The research in this R56 project will involve computer simulations and experimental tests using the HTS head-only MRI scanner operating at 0.7 Tesla. We will develop a multichannel RF coil and multi-echo 2D-FREE imaging with parallel RF transmission and reception. Products will include new MRI methods, software, and hardware to achieve highly portable midfield MRI. Future portable mid-field MRI scanners based on this new technology will help people in remote, resource-limited settings to address health inequities.

NIH Research Projects · FY 2025 · 2024-09

Abstract Freezing of gait (FOG) is characterized by episodes during which an individual is unable to step, despite intending to do so, and is a common cause of falls, decreased mobility, and increased morbidity in people with Parkinson’s disease (PD). Effective treatment for FOG remains elusive, due to a lack of understanding of the complex underlying pathophysiology. The expression and factors contributing to FOG are highly heterogeneous across individuals, however, a common feature is that episodes are predominantly triggered during movement-state transitions (e.g. initiating walking or turning)4,5. Impaired transitions leading to FOG typically occur when the change in movement-state is self-initiated (uncued), but when the same transition is cued by an external sensory stimulus, movement execution is improved, and the incidence and duration of FOG is markedly reduced. FOG may be caused by abnormal communication between subcortical systems controlling posture and balance (e.g. vestibulo- and reticulospinal systems) and cortico-fugal systems driving the initiation of the intended action (e.g. stepping). Currently, the mechanisms contributing to impaired self-initiated movement transitions in people with PD and FOG, and how they are improved by external cues, are poorly understood. We hypothesize that the capacity to downregulate the communication (coherence) of systems controlling posture and balance during self-initiated transitions from one movement state (standing) to another (walking) is impaired in people with FOG, and that sensory cues ameliorate FOG by restoring transition-related modulation of posture/balance systems. This hypothesis will be tested using biomechanical and neurophysiological measures to examine the dynamics of the vestibulo-postural (Aim 1), cortico-cortical and cortico-muscular (Aim 2), and cortico-basal ganglia (Aims 3 and 4) systems during cued and uncued posture-locomotion transitions in PD, with and without FOG, and controls. Aim 1 will examine the vestibulo-postural system in FOG by measuring the dynamic changes in coherence between vestibular input (electrical vestibular stimulation) and the ground reaction forces controlling balance. Aim 2 will utilize high-resolution electroencephalography (EEG) and electromyography (EMG) to examine movement-related cortical potentials and cortico-cortical and cortico-muscular coherence. Aim 3 will use EEG and local field potential (LFP) recordings from implanted Medtronic PerceptTM deep brain stimulators (DBS) to examine the interaction of the globus pallidus and cortex (cortico-pallidal coherence). Aims 1-3 will utilize standardized gait initiation paradigms that may or may not provoke freezing. Aim 4 will use wearable sensors and a FOG-provoking course, involving multiple posture/gait transitions, to wirelessly capture LFPs associated with FOG episodes in participants with PD with the PerceptTM DBS system. This project will provide insight into the mechanisms and neurological biomarkers of FOG, which will be critical for the development of interventions to reduce the incidence and severity of FOG, reduce falls, and improve the quality of life of people with PD.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Diabetes is a heterogeneous disease characterized by chronic poor glycemic control, resulting in pathological changes in tissues throughout the body. Chronic hyperglycemia causes the cardiovascular system to undergo growth and remodeling (G&R) that is captured hypertension and increased arterial stiffness, both significant risk factors for cardiovascular disease. Diabetes results in both cellular and matrix changes in arterial health, and then each of these components further drive the pathological G&R by responding to the direct effects of the disease. Fortunately, there exist medical and lifestyle interventions to help mitigate the diabetic disease state and restore glycemic control. Large elastic arteries like the aorta serve as capacitors to absorb changes in blood volume due to pulsatile pumping and protect the more fragile downstream microvasculature. Overall health and stiffness of these large vessels is captured in the clinic via pulse wave velocity which is elevated in diabetic patients, and there is evidence that the pulse wave velocity decreases back towards baseline following restoration of glycemic control. The overarching objective of this project is to determine how hyperglycemia affects aortic biomechanics and mechanobiology and whether restoration of normoglycemia is sufficient to reverse these changes. To establish how arterial biomechanics are affected by hyperglycemia, I first employed a diabetic mouse model to ascertain changes in active and passive wall mechanics, and these preliminary results demonstrate that chronic hyperglycemia results in stiffer and hypercontractile murine aortas. The central hypothesis of this proposal is that chronic hyperglycemia results in cellular and matrix aortic G&R, and glycemic recovery results in reversal of the cellular, but not matrix, phenotype, leading to partial rescue of aortic health. To test this hypothesis, I will utilize an inducible mouse model of chronic hyperglycemia that can subsequently be rescued by administration of Phloridzin. Experiments proposed in Aim 1 will determine the in vivo and ex vivo biomechanical changes in aortic health due to hyperglycemia and following treatment. Aim 2 will then explore how the tissue-scale mechanical changes arise by investigating how the matrix composition and cellular phenotype are affected by the disease and treatment. The experimental work of the first two Aims will be coupled with a multiscale, bio-chemo-mechanical computational model of aortic G&R in Aim 3. The computational model will provide mechanistic insight into the roles of cells and matrix in disease progression and possible regression, resolving information that is inextricable experimentally. Collectively, these data will elucidate aortic G&R due to chronic hyperglycemia, whether these changes can be reversed by restoration of glycemic control, and the mechanisms behind these processes. This proposed work will have broad implications in the conceptual understanding of the cellular and matrix roles in diabetes-related cardiovascular pathologies and the development of methodologies to explore biomechanical and mechanobiological changes in vascular health due to disease and treatment.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Species exhibit stereotypic lifespans, suggesting an intrinsic hardwired limit. Our preliminary data shows that a population of mouse T cells can be iteratively boosted in vivo and passaged among young mice for >11 years (~4 mouse lifespans). This multi-lifespan T cell population remains functional, does not undergo unrestrained cell division, and like young memory T cells, undergoes rapid proliferative bursts upon further antigen/mitogen stimulation. We have now generated 10 indepdent ongoing cohorts of cells, ranging from 6-66 total boosts over 1-11 years, to study mechanisms by which functional cells can exceed their species’ lifespan while retaining cell identity, functional and proliferative competence, accumulating perhaps 100s or even 1000s of cell divisions, while not exhibiting signs of uncontrolled proliferation. Specific aims will reevaluate the Hayflick Limit, assess mutational burden and the accumulation of transcriptional noise, test whether a young host environment is responsible for maintaining everlasting functional persistence of transferred T cells, and evaluate metabolic adaptations that may support extreme longevity and maintenance of proliferative potential. Our long-term goal is to understand how a population of mammalian somatic cells has adapted the capacity to maintain vitality despite multi- lifetime chronological aging and an excessive history of proliferation, and to ultimately apply newly learned concepts and mechanisms to extend human health span. .

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Cardiomyopathy is classified into ischemic cardiomyopathy, caused by coronary artery disease, or non- ischemic cardiomyopathy, caused by other factors. The current standard practice for diagnosing the cause of cardiomyopathy is to use coronary angiography to assess the presence or absence of coronary artery disease. However, this approach is flawed and may not always accurately identify the underlying cause of the myocardial disease. Recent advancements in cardiac magnetic resonance imaging allow for a myocardial approach to identifying the cause of cardiomyopathy by directly visualizing myocardial abnormalities, including tissue damage. We hypothesize that a myocardial approach will reclassify the cause of cardiomyopathy in a clinically significant proportion of patients and improve the prediction of long-term outcomes. To test this hypothesis, we will use a large cohort of patients from an institutional registry of consecutive patients undergoing cardiac magnetic resonance imaging for clinical reasons. Our study's anticipated results will provide novel, large-scale data on the cause of cardiomyopathy and its long-term prognostic impact, clarifying the extent of misclassification when using a coronary approach. This information will be essential to conduct a randomized clinical trial comparing a myocardial approach to the coronary approach for identifying the cause of cardiomyopathy. More accurate recognition of the cause of cardiomyopathy will allow for increased use of appropriate cause-specific treatments, improving the long-term outcomes of patients. The study's results will be relevant to the 1 million Americans diagnosed with heart failure and the several thousand others diagnosed with cardiomyopathy without heart failure every year. The proposed work has a high potential to lead to improvements in clinical practice and patient outcomes on a large scale.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Cells shape and reshape themselves as they accomplish diverse functions in vivo. These cell shape changes result from physical and chemical feedback loops at multiple scales. At the biochemical scale, spatiotemporally varying signaling cascades control cytoskeletal polymerization that pushes on the plasma membrane from the inside, inducing cell morphology change at the scale of micrometers. Cell morphology in turn feedbacks to alter intracellular signaling, via mechanisms such as surface curvature-sensing proteins in the plasma membrane. Understanding these feedbacks across scales is particularly critical since they are a target for pharmaceutical intervention. Scientists have developed a wealth of techniques for interrogating the biochemical and genetic basis of cell function. However, disentangling the many feedback loops coupling cell morphology, intracellular organization, and spatiotemporally varying biochemical pathways, such as intracellular signaling, remains ex- perimentally challenging. Live-cell fluorescence microscopy can visualize these feedbacks in action, but imaging across scales produces enormous and detailed datasets that are impossible to make sense of without dedicated computational pipelines. We will develop algorithms for cell biology rooted in computational geometry. Using computational geometry approaches will allow us to draw from decades of math and computer science research to work in biologically relevant geometries, increasing accuracy and easing interpretation. We propose to develop essential algorithms to reconstruct plasma membrane organization from 3D microscopy images and track its movements across time. We will also measure the organizational rules that couple plasma membrane shape and dynamics to spatiotemporally varying biochemical pathways on the cell surface. Finally, we will focus on understanding feedbacks related to intracellular organization throughout the cell, not just on the plasma mem- brane, by broadening the computational approaches we developed for the plasma membrane to other geome- tries. The computational methods that we are proposing to develop will aid in further opening up to scientific investigation the many feedback loops and physical interactions of the meso-scale world of subcellular organi- zation.