University Of Minnesota

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Atrial fibrillation is the most common sustained arrhythmia and its prevalence is increasing. Accumulating evidence suggests that upstream to atrial fibrillation, structural, mechanical, or electrophysiological changes in the atria—collectively known as atrial myopathy—may be a primary mediator of heart failure, ischemic stroke, and mortality in patients with atrial fibrillation. However, the pathophysiology of atrial myopathy is incompletely understood, there are currently no targeted therapies, and treatment targets suggested by human population studies remain unvalidated experimentally. Therefore, treatment target validation and understanding the pathophysiologic mechanisms that contribute to atrial myopathy are unmet needs. This project aims to fill this gap by identifying novel treatment targets through large-scale human data analysis and by investigating the protective role of the protein DLK1 in atrial myopathy. The primary aim leverages human cohort data to identify candidate treatment targets by discovering circulating proteins and metabolites linked to both left atrial function and atrial myopathy. By combining traditional multivariable linear regression analyses with advanced genomics-based causal inference methods, we aim to prioritize promising targets for further preclinical investigation, ultimately paving the way for novel atrial myopathy therapies and improved clinical atrial fibrillation management. Because prior studies suggest DLK1 protects against fibrosis and inflammation and our preliminary data found that higher levels are associated with better left atrial function. The secondary aim of this project will investigate DLK1's role in mouse models of atrial myopathy, testing if increasing atrial DLK1 expression improves left atrial function and if decreasing atrial DLK1 expression impairs left atrial function. Aiming to become an independent physician-scientist in atrial fibrillation and atrial myopathy research and to bridge population and basic science, the applicant seeks K08 funding for protected time to solidify his skillset in characterization of cardiac fibrosis, interrogation of atrial pathophysiology, and Mendelian randomization for causal inference. This protected research time will give the applicant an opportunity to build his research program, gather preliminary data, and ultimately secure R01 funding to establish himself as an independent investigator in the cardiovascular field. The applicant has also assembled an accomplished and well-funded multidisciplinary mentoring team, comprised of internationally known experts in the fields of atrial fibrillation and atrial myopathy epidemiology, cardiomyocyte biology, cardiac fibrosis and diastolic dysfunction, and Mendelian randomization. With their support and guidance, and the research infrastructure available at University of Minnesota, the applicant is well-positioned to achieve his proposed research aims and accomplish his stated career development goals. The applicant’s clinical expertise in multi-modality cardiovascular imaging aligns perfectly with his research goals, enabling him to investigate novel biomarkers and translate research findings to improve patient care.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Sexually reproducing organisms use meiosis to distribute one copy of each homologous chromosome pair to developing gametes. This relies on the formation of physical linkages between chromosomes known as crossovers, and crossover errors are a leading cause of infertility and conditions such as Down syndrome. Accordingly, while crossovers appear at random genomic positions in each meiosis, they are tightly regulated to ensure each chromosome pair receives at least one crossover and that crossovers beyond the first are well- separated along the length of the chromosomes. Yet while these phenomena were first observed over a century ago, the mechanism by which crossover locations are coordinated along chromosomes remains hotly debated. To investigate this longstanding question, our lab recently pioneered the use of single-molecule imaging to track the motion of proteins known to regulate crossover formation. We found that these proteins are recruited by and move along a nanoscale protein assembly between the chromosomes known as the synaptonemal complex, allowing them to coordinate their activity at sites millions of base pairs apart. Our current research program works to understand how dynamics in this liquid-like compartment create emergent patterns of crossovers. In our first major research direction, we will study the proteins required for crossovers to form. One leading theory of crossover patterning proposes that the distribution of crossovers along chromosomes emerges from competition between sites to recruit enough of these proteins to form crossovers. To explore this possibility, we will evaluate how quickly and how far different proteins can move along chromosomes, how proteins bind to potential crossover sites, and what regulates these dynamics. Using our precise measurements of diffusion and binding kinetics, we will use computer modeling to understand how these dynamics collectively generate tightly regulated crossover numbers and spacing, and how mutations in these proteins disrupt regulation. In our second major research direction, we will study the dynamic properties of the synaptonemal complex itself. As a liquid-like assembly, small genetic perturbations or changes in environmental conditions can alter the material properties of the synaptonemal complex and drastically impair fertility. We will study how structural changes and shifts in temperature alter the function of the synaptonemal complex, as well as how the synaptonemal complex in turn is patterned and internally organized by the formation of crossovers. Altogether, our research program will uncover fundamental mechanisms of how crossovers are regulated and how molecular activity along chromosomes is spatially patterned. The insights provided from this research are critical for understanding how infertility and birth defects arise in humans and how liquid-like compartments organize the nucleus, with implications for related diseases such as errors in DNA repair in cancer.

NIH Research Projects · FY 2025 · 2025-09

Project Summary / Abstract Despite widespread use of buprenorphine to treat opioid use disorder (OUD), 50% of patients return to use within months. Non-invasive prefrontal stimulation is a highly promising buprenorphine augmentation strategy to enhance prefrontal plasticity to boost executive functioning (EF) needed for recovery. We reported that a neuromodulation intervention combining prefrontal stimulation and EF training delivered to individuals with alcohol use disorder increased prefrontal-incentive salience network connectivity and resulted in a significant reduction in relapse rates. This intervention has not yet been tested as a buprenorphine augmentation strategy for OUD. This proposal investigates the added benefit of the intervention in individuals under buprenorphine maintenance treatment for OUD (bOUD) to determine whether this intervention can: (i) induce circuit-based target engagement, (ii) enhance EF, and (iii) improve treatment outcomes. UG3 phase, open-label, within- subjects. Deliver 10 days of active transcranial direct current stimulation (tDCS) to individuals with bOUD concurrent with cognitive training. Brain imaging data and EF performance will be collected at pre-, mid- and post-intervention. UG3 aims to determine if there is a dose-dependent tDCS effect by contrasting change from baseline after 5 vs after 10 tDCS sessions in the following metrics: (SA1) circuit-based target engagement of a prefrontal-incentive salience network associated with abstinence and (SA2) EF improvement, i.e. working memory, cognitive flexibility, inhibitory control, decision making. SA3 will involve a pre-submission meeting with the FDA and NIH to review and obtain feedback on: regulatory process, UH3 design and study endpoints. UH3 phase, randomized-controlled trial, between-subjects. With FDA feedback and UG3-determined dose, a between-subjects, triple-blind, randomized-controlled clinical trial will assign 120 individuals with bOUD to either active tDCS or sham concurrent with cognitive training. We will collect brain imaging and EF changes at pre- and post- intervention, treatment outcome measures at 1-, 3- and 6-month follow-up timepoints. UH3 aims to contrast active tDCS vs sham groups on (SA4) clinical efficacy and durability of active tDCS on treatment outcome (craving, opioid use) over the 6-month follow-up period. To examine intermediate factors playing a key role in the intervention’s mechanism of action, we will (SA5) determine the relationship between changes in target engagement, EF and treatment outcomes. The UH3 is positioned to be in Phase 2 of the FDA Regulatory Pathway, to design either a Phase 2 or 3 Clinical Trial to support a Premarket Approval application. Impact: We propose the first study to examine the added benefit of a non-invasive prefrontal neuromodulation intervention designed as a highly promising buprenorphine augmentation strategy to dually target executive functioning deficits and excessive craving in individuals with bOUD. Success in these aims would provide crucial information about this combined intervention, setting the stage for clinical translation of a low-cost, scalable, neuroscience-based treatment for OUD.

NIH Research Projects · FY 2025 · 2025-09

The National Institute on Aging Demography and Economics Coordinating Center (NIA DECC) will help the NIA P30-funded Demography and Economics of Aging and AD/ADRD (D&E) Centers realize their full potential to generate impactful research, to facilitate new and innovative collaborations, and to broaden the community of aging and AD/ADRD scholars. NIA’s investment in a Coordinating Center is only justified if it increases the efficiency, productivity, and impact of the collective group of D&E Centers beyond what they can accomplish in isolation. We propose an exciting, interactive, and dynamic new vision for what the D&E CC can do for the D&E Centers, for NIA, and for the community of researchers worldwide who will leverage our resources to shape the next generation of research on the demography and economics of aging and AD/ADRD. We will do this by (1) serving as the administrative hub of the D&E Centers; (2) facilitating synergistic collaborations to catalyze impactful aging research; (3) expanding the impact of the D&E Centers’ research through effective dissemination; and (4) coordinating tracking and reporting to reduce redundant effort and increase productivity. The suite of activities we propose leverages the many strengths, assets, expertise, and programming of the University of Minnesota’s Institute for Social Research and Data Innovation to coordinate and enhance the activities of the individual D&E Centers; the objective is to make the whole more productive and impactful than the sum of its parts. The NIA DECC will create and support a vibrant nationwide community of aging and AD/ADRD scholars that is better informed, supported, and connected to address urgent aging-related problems. Like the individual D&E Centers and NIA, we are also deeply committed to growing the pool of scientists studying the D&E of aging and AD/ADRD. We will coordinate, organize, and catalyze the individual D&E Centers’ efforts to recruit and support Emerging Scholars. The NIA DECC will be led by an interdisciplinary MPI team that has, for more than a decade, collaborated to successfully develop, implement, grow, and sustain multiple research centers, major research projects, training programs, professional development programs, and initiatives to help early career scholars develop their research agendas and funding portfolios. We envision the NIA DECC as a central hub of innovation, information exchange, and new ideas and collaborations, connecting the D&E Centers to one another, to NIA, to other NIA P30-funded networks (e.g., the Roybal Centers, the Alzheimer’s Disease Research Centers), to policymakers, and to the public. The proposed activities will increase the productivity of all the D&E Centers and grow and broaden the aging and AD/ADRD research community, supporting the P30 Center Program’s goal to advance aging research in demography, economics, and related interdisciplinary population-based social science areas, including those with a focus on AD/ADRD.

NIH Research Projects · FY 2025 · 2025-09

Program Director/Principal Investigator (Last, First, Middle): Naselaris, Thomas PROJECT SUMMARY We propose to amass a large sampling brain activity as humans internally generate mental images. The proposed Mental Imagery Database (MID) will use high-resolution 7T fMRI to achieve large-scale sampling of brain activity during mental imagery, a timely project directly motivated by the rise of generative AI. Of the many use cases we envision, we see two as especially urgent: (1) MID will be used by neuroscientists and AI researchers to gain insights into how the brain transforms text into novel visual representations. This operation is central to current AI research and distinguishes humans from other biological intelligences. Although AI image generators have become better at making pictures than most humans, they still struggle with visual interpretations of written information that humans find easy. Very little is currently known about how human brains do this. MID will provide the needed neuroimaging data at a scale that is matched to the complexity of the problem. (2) MID will advance recent work on visual decoding that has made great strides reconstructing seen images. Such work is hampered by the lack of data for cross-generalization to mental imagery. Using MID to fine-tune vision decoders for mental imagery would bring us closer to viable technology for externalizing visual thoughts. This technology could be deployed by clinicians to help patients interrogate and control intrusive mental images of traumatic events, and aid diagnosis and communication for unresponsive patients whose consciousness is not readily apparent through standard behavioral assessments. Decoding such patients' mental images could, in principle, affirm consciousness and aid in accurate diagnoses. RELEVANCE We expect this work to deliver a method for producing high-quality reconstructions of visual mental images. This work will impact the diagnosis and treatment of mental health disorders that are driven by dysregulated visual mental imagery. Little is known about the brain systems that mediate mental imagery's role in mental health, in part because of the paucity of high-quality mental imagery data, which the proposed Mental Imagery Database will provide.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Traumatic brain injury (TBI) is a leading cause of death and disability, and is associated with elevated risk of chronic health conditions including dementia. Cerebral edema is a particularly deleterious complication of TBI which greatly increases risk of death, but current clinical approaches for edema management do not lead to signif- icant improvements in patient outcomes. Recent translational studies of TBI in mice demonstrate that trigeminal nerve stimulation (TNS) offers a low-risk, noninvasive therapeutic approach that reduces edema and improves functional outcomes. However, stimulation parameters vary widely across studies, and the mechanisms of edema reduction are not well-understood. The research proposed herein seeks to utilize both mouse experiments and realistic numerical simulations to probe stimulation parameter sensitivity and obtain a mechanistic understanding of how nerve stimulation leads to edema reduction. Such insights are necessary for rapid and effective trans- lation of TNS-based therapies, and indeed, results from this research will help inform upcoming TNS studies in humans. The central hypothesis of this proposal is that TNS improves functional outcomes after TBI by enhanc- ing extracellular fluid transport through the glymphatic system (a pathway in which cerebrospinal fluid exchanges with interstitial fluid in the brain). Four lines of evidence support this hypothesis: (1) a recent Nature publica- tion demonstrates that restoring disrupted glymphatic transport following TBI removes excess fluid and cortical debris, sharply reducing neuroinflammation and improving functional outcomes; (2) glymphatic flow is driven by arterial pulsations, and prior TNS studies use pulsed stimulation that enhances arterial pulsatility; (3) prior work shows that stimulation of the vagus nerve enhances glymphatic transport; and (4) multiple studies show analo- gous sensory (e.g., whisker) stimulation alters cortical blood flow and enhances glymphatic transport. Specific Aim 1 will test the central hypothesis by measuring brain-wide glymphatic influx due to variable TNS parameters (stimulation frequency, intensity, duty cycle) and will yield a near-optimal parameter set corresponding to maxi- mum glymphatic transport. Specific Aim 2 will provide the first high-resolution, in vivo quantification of changes in arterial diameter and glymphatic transport during TNS. This will be achieved using two-photon microscopy while TNS is administered with near-optimal parameters. Specific Aim 3 will leverage an existing, versatile simulation of the murine glymphatic system to causally establish the extent to which increased arterial pulsatility enhances glymphatic transport. Results from this study will: (1) conclusively demonstrate whether TNS ameliorates acute post-TBI edema via enhanced glymphatic transport, (2) yield a set of near-optimal TNS parameters, (3) directly quantify TNS-induced alterations to blood/glymphatic flow, and (4) quantify the extent to which increased glym- phatic transport due to sensory/nerve stimulation may be attributed to increased arterial pulsatility. Importantly, this study will have direct translational value: research results will help guide upcoming human studies in which TNS will be administered to humans using a noninvasive, wearable device with potential for commercialization.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract My laboratory works at the interface of chemistry and biology on technology development for post-translational modification enzyme activity measurement. We use a broad variety of techniques including peptide chemical biology, spectroscopy, proteomics, cell biology, and analytical chemistry to create tools that help answer biological questions with molecular precision. We have spent the most time on studying and building a toolkit for measuring kinase activity, both in vitro and in live cells. We are currently working on building out our platform technology called KINATEST-ID to characterize kinase preferences, design novel substrates, and apply them in kinase assays with innovative read-outs. Our goals for the next five years are (1) to build up our understanding of kinase-substrate interactions using multiple avenues: in vitro phosphoproteomics experiments to characterize the substrate profiles of many different kinases, comparing substrate preference motifs to try to dissect differences, designing novel artificial substrate tools that could distinguish between the activities of different kinases, and using cutting-edge artificial intelligence-based structure-based computational methods like Rosetta and AlphaFold to characterize and predict determinants of substrate selectivity; and (2) to develop and implement assay read-out detection methods that can meet different levels of technical needs for drug screening, drug target validation, pharmacodynamic monitoring for companion diagnostics, as well as basic study of fundamental questions about enzyme function and interaction with substrates in cells.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract: The evolution of antimicrobial drug resistance is an urgent threat to public health. A better understanding of basic evolutionary principles in microorganisms can inform efforts to identify, prevent, and combat the emergence of drug resistance. Candida albicans, one of the most common fungal pathogens, can adapt to antifungal drugs via mutations that alter gene expression levels. Gene expression levels can be altered by changes in gene copy number and mutations that alter transcriptional regulatory networks. Basic science investigating the influence of gene copy number changes on fitness and the impact on the gene regulatory network across diverse genetic backgrounds is needed. These investigations will directly contribute to combating the emergence of antifungal drug resistance. The objective of the proposed research is to test the effects of copy number changes on fitness and gene regulatory networks, and relate these effects to the rate and dynamics of the evolution of antifungal drug resistance in C. albicans. In the long-term, this work can be extended to additional fungal pathogens and related to clinical sampling data, as well as inform general evolutionary biology principles that can be tested and applied across the tree of life. Aim 1 uses newly developed CRISPR-activation technology in C. albicans to test the fitness effects of gene overexpression in multiple genetic backgrounds and environments. These data will be used to describe the distribution of fitness effects of recurrently amplified genes. Aim 2 combines CRISPR- activation technology with single-cell RNA-sequencing to model transcriptional regulatory networks. These will be used to identify genome-wide transcriptional effects of gene amplification in diverse genetic backgrounds and environments. Aim 3 uses experimental evolution to test the hypothesis that a gene’s position within the regulatory network influences the rate and dynamics of adaptive gene expression change in resistance genes. The long-term career goals of the candidate are to establish a research program centered around genetic constraints and facilitations to evolutionary adaptation at a major biomedical research institution. The short-term training goals of this proposal are to further develop the candidate’s expertise in the computational representation of regulatory networks and in the application of single-cell RNA-sequencing technologies to pathogenic yeast. The University of Minnesota provides an ideal environment in which to pursue these training goals, with excellent resources for career development, extensive genomics and computing core facilities, and faculty with expertise in genomic networks, evolutionary biology, and microbiology that will serve as mentors and advisors to the candidate. The training and research proposed will launch the candidate into an independent and productive research career.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Dopamine (DA) is a critical neuromodulator for mood, motivation, and reward-based learning. Dysregulated DA dynamics across brain regions and timescales contribute to psychiatric and substance use disorders, yet how DA timescales mediate the relationship between learning and motivation remains unresolved. Striatal DA release is documented to fluctuate on fast “phasic” and slower “tonic” timescales, hypothesized to regulate learning and motivation, respectively. Prevailing reinforcement learning (RL) theories suggest that phasic DA transients encode reward prediction errors (RPEs), which diffuse and accumulate into tonic DA levels to regulate motivational engagement. This interpretation assumes that subjective values driving motivational engagement are reactive to recent reward exposure. However, alternative theories propose that tonic DA may arise independently from phasic DA, reflecting proactive effort allocation based on internal calculations rather than recent reward exposure. Resolving these conflicting views is essential for understanding DA’s role in motivation and its dysfunction in psychopathology. This proposal aims to define the circuit and computational mechanisms linking phasic and tonic DA to flexible motivational engagement. Using a novel broadband DA measurement technique we recently developed, I will analyze how DA dynamics across timescales interact to support reactive-vs-proactive motivational flexibility. Aim 1 will assess how these valuation processes influence the relationship between phasic and tonic DA. Aim 2 will identify circuit mechanisms regulating how phasic DA signals accumulate into tonic levels under varying task demands of reactive-vs-proactive value learning. These studies will reconcile longstanding theoretical debates on DA function, providing a mechanistic and computational framework for understanding how DA shapes motivation and learning across timescales. The insights gained from these experiments will inform targeted interventions for circuit-specific vulnerabilities in addiction, depression, and psychosis.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Cells are collectively oriented along tissue axes to execute coordinated behaviors during embryonic development. The highly conserved Planar Cell Polarity (PCP) pathway allows cells to communicate directional information with each other to direct essential processes such as neural tube closure, axis elongation, and heart morphogenesis. The core PCP pathway includes three transmembrane proteins (Celsr1, Fzd6, and Vangl2), which form complexes on opposite sides of the cell. Through interactions with intracellular binding partners, these proteins are able to self-organize such that Vangl2/Celsr1 complexes enrich on one side while Fzd6/Celsr1 complexes enrich on the other side of the cell. Extracellularly, Vangl2/Celsr1 complexes form an asymmetric junction with Fzd6/Celsr1 complexes to directly link PCP asymmetry between neighboring cells. A defining characteristic of PCP organization is its alignment with a tissue axis which allows cells to uniformly orient PCP asymmetry across great distances. This stereotypic pattern cannot be achieved by self-organization alone as cells could spontaneously organize in any orientation with respect to the tissue axis. For this reason, a well-accepted hypothesis emerged that a ‘directional cue’ acts in a gradient across a tissue to bias the distribution of PCP proteins along the same axis. Currently, the identity of potential ‘directional cues’ remain elusive, and how the core PCP pathway interacts with the cue to generate tissue-level asymmetry is unknown. Forward genetic screens are a powerful tool to identify genes involved in a particular process based on the phenotype. In fact, core PCP genes were identified in Drosophila based on disrupted bristle orientations in the wing and thorax of PCP mutants. However, the same approach is less successful in identifying genes that encode ‘directional cues’ as they are also thought to be essential for the development of the tissue itself. Fortunately, breeders have performed their own forward genetic screen in guinea pigs and mice, selecting for naturally occurring genetic variants with altered hair follicle orientation. Similar to Drosophila bristles, mammalian hair follicles are oriented by the PCP pathway. The altered fur orientation in these natural variants shows that planar polarity alignment is decoupled from the body axis in a region-specific and uniform manner, suggesting the causative mutations alter ‘directional cues.’ Remarkably, the viability of the animals and the overall architecture of the skin itself remain unaltered. By turning to these natural variants, I hypothesize that I will be able to identify ‘directional cues’ that link PCP asymmetry to a tissue axis. By computationally mapping the genetic variants, I will reveal genes required to link PCP alignment to a tissue axis. Through phenotypic characterization in vivo and in vitro, I will define how the altered gene impacts PCP asymmetry at individual junctions and its alignment across the entire tissue. This research will uncover mechanisms coordinating cell behaviors across tissues, lending insight into a fundamental process that is required for proper development.

NIH Research Projects · FY 2025 · 2025-08

Project Summary How does mutant ATXN1 expression in microglia impact Spinocerebellar Ataxia Type 1 pathogenesis? Spinocerebellar ataxia type 1 (SCA1) is a fatal autosomal dominant neurodegenerative disease without a cure or effective therapies to delay disease onset and progression. SCA1 is caused by a CAG repeat expansion that encodes for a polyglutamine stretch in the ATAXIN-1(ATXN1) protein and is characterized by deficits in motor coordination, dysarthria, and premature death. Prior work has uncovered increased numbers of microglia, microglia contribution to SCA1 motor phenotypes, and differentially expressed genes in SCA1 microglia. However, there is still a critical need to understand whether these phenotypes are due to mutant ATXN1 (mATXN1) expression in microglia or surrounding cells. The goal of my proposal is to determine the role microglial mATXN1 expression plays in motor behavior, microglia molecular phenotypes, and the impact on surrounding cells. I aim to achieve this by removing mATXN1 from microglia by crossing Lyve1CRE mice with f- ATXN1146Q/2Q mice. The f-ATXN1146Q/2Q mouse model contains LoxN sites around mATXN1 allowing for conditional deletion of mATXN1 with Cre recombinase expression. The Lyve1CRE model highly expresses Cre recombinase in microglia and macrophages. Crossing these two mice will generate f-ATXN1146Q/2Q;Lyve1CRE mice which will have mATXN1 deleted from microglia and macrophages. I aim to assess the impact of removing mATXN1 expression in microglia on motor deficits, microglia disease phenotypes, and cerebellar pathogenesis. Motor function will be assessed through battery of motor assay. Microglia disease phenotypes and cerebellar pathogenesis will be assessed with a combination of single nuclei and bulk RNA sequencing and immunohistochemistry. I hypothesize that removal of mATXN1 will ameliorate motor deficits, correct the number of differentially expressed genes in microglia and other cerebellar cells, and ameliorate microglial reactivity and cerebellar pathology seen through immunohistochemistry. This research will aid in further elucidating the role microglia plays in SCA1, a relatively understudied aspect of SCA1. Finally, the training I will receive during this work is paramount to my goal to become faculty at research university.

NIH Research Projects · FY 2025 · 2025-08

Abstract The long-term, overall mission of this T32 is to provide a comprehensive and multi-level training program, exposing trainees to transdisciplinary science, and providing the building blocks to launch a long-lasting career that address pressing issues in musculoskeletal research. Within this five-year program, we will focus on the concept of interdisciplinary musculoskeletal research. Specifically, the proposed training program will provide integrated training for five predoctoral students in our three thematic areas: 1) MSK Development and Aging; 2) MSK Imaging and Translational Studies; and 3) MSK Cancers and Osteoimmunology. Our selected faculty within these themes align with the central concept of interdisciplinary research and have a strong track-record of mentoring predoctoral students. Our selected faculty within these themes align with the central concept of interdisciplinary science and have a strong record of accomplishment of mentoring predoctoral students and cross-disciplinary collaboration. Faculty within the MSK Development and Aging theme focus on understanding how mechanisms governing correct formation of MSK tissues and how these processes go astray with age. Early detection of MSK conditions aids in the management of MSK conditions, which is the focus of faculty within the MSK Imaging and Translational Studies theme. Lastly, faculty within the MSK Cancer and Osteoimmunology theme aim to identify potential treatments for MSK cancers and explore how MSK tissues, including cancers, interface with the immune system. Along with these topical foci, our training program incorporates methods to achieve specific training goals, including: 1) exposing trainees to broad MSK research topics and methods that address MSK conditions; 2) supplying training on ethics and reproducibility within research; 3) delivering skills needed to develop independent lines of research; and 4) providing networking and career development opportunities. This training program will ensure that trainees from diverse backgrounds will be in the best possible position to contribute effectively to advancing MSK science.

NIH Research Projects · FY 2025 · 2025-08

Most phenotypic traits, including nearly all morphological, physiological, and molecular quantities, vary continuously among individuals in a population. These traits are shaped by DNA variation at dozens to thousands of genes, giving them a complex, multigenic basis. Over the past 20 years, there has been remarkable progress in identifying genomic regions that are associated with complex traits. However, few of the causal genes that shape complex traits have been identified. This proposal addresses this key problem by introducing a new method for genetic mapping called CRI-SPA-Map. Our objectives are to apply CRI-SPA-Map in three distinct, complementary contexts in the yeast Saccharomyces cerevisiae: a pair of closely related strains, a pair of more distantly related strains, and between S. cerevisiae and its reproductively isolated sister species S. paradoxus. In each of these contexts, we will identify and analyze causal genes that shape the ability of yeast strains to grow in a set of diverse environmental conditions. CRI-SPA-Map makes it possible to transfer small tracts of DNA from one yeast strain to a genetically different strain efficiently, in high throughput, and with minimal cost. Combined with high-throughput phenotyping, CRI-SPA-Map enables systematic discovery of causal genes that shape complex traits. We anticipate that our proposed activities will result in discovery of dozens to hundreds of causal genes. Systematic analyses of these genes will provide new insights into the genetic architecture of complex multigenic traits, such as the precise number and effects of causal genes, how their functional roles relate to the given trait, and how their effects vary across conditions. This proposal will also generate a community resource of libraries of allele-engineered and whole-genome sequenced yeast strains, allowing mapping of any trait of interest to gene-level resolution. We will pursue two specific activities in support of the broader impacts of this proposal. In support of improved STEM education, this grant will support a teaching experiment to test if the order in which genetic concepts are taught can improve student understanding of multigenic traits. In support of the development of a competitive STEM workforce, this proposal will establish a new pathway for undergraduate research experiences in complex trait genetics. RELEVANCE (See instructions): Most phenotypic traits are shaped by DNA variation at dozens to thousands of genes, but few of the causal genes have been identified. This proposal addresses this key problem by introducing a new method for genetic mapping, which we will apply to reveal causal genes in three contexts in the yeast Saccharomyces cerevisiae. Systematic analyses of these genes will provide new insights into the genetic architecture of complex multigenic traits.

NIH Research Projects · FY 2025 · 2025-08

Abstract Monitoring of ascending thoracic aortic aneurysm by contrast CT presents major cost, health, and patient availability issues, especially for patients in rural areas. Although CT is expected to remain the gold standard for patient monitoring, these issues make it attractive to explore an alternative monitoring modality that could be done more often, possibly even at home, and could be used to screen whether the full CT is necessary. Such a tool would not only help reduce the number of CT scans needed for stable patients, it would also have the potential to be used more often than the usual 6-12 month frequency of surveillance CT, meaning that a potential problem could identified sooner. In this exploratory project, we will determine whether carotid pulse arrival time (cPAT) has the potential to be such a monitoring tool. cPAT is easily measured via surface tonometry, and electrical-impedance-based methods are even simpler and less costly. cPAT is also a direct measurement of flow through the ascending thoracic aorta, unlike the standard pulse wave velocity measurements that test wave speed between, e.g., the carotid and femoral arteries. The appeal of cPAT is clear, but it is not clear that the measurement is sensitive enough to measure significant changes in the ascending aorta. Thus, we will use two Specific Aims to assess the potential of cPAT as a monitoring tool. First, we will perform an experiment on a large cross-sectional cohort to determine whether cPAT results are consistent with the empirical expected pulse wave velocity based on age and blood pressure, testing whether cPAT can measure a population-level trend that one would expect to see. Second, we will perform computer simulations of fluid-structure interaction flow in the ascending aorta, using realistic geometries based on patient scans. These simulations, which will include longitudinal patient scans and artificially enlarged aneurysms, will allow us to assess how much of a change in vessel properties could be detected given the time resolution of the cPAT. Together, these two Aims will determine whether a larger-scale study is merited on whether cPAT can be an effective monitoring tool.

NIH Research Projects · FY 2026 · 2025-08

ABSTRACT Somatic mosaicism arising from the accumulation of postzygotic DNA mutations in somatic cells (somatic mutations), leads to tissue genetic heterogeneity. Somatic mutations occur at different life stages and throughout an individual’s lifespan and are implicated in various human diseases, including cancers, monogenic diseases, neurodegenerative disorders, cardiovascular diseases, autoimmune diseases, skin diseases, and liver diseases. In addition, random somatic mutations were identified to accumulate with age in humans, and associated with environmental and genetic risk factors, for example, tobacco smoking, UV radiation and BRCA1/2 deficiency. However, it is still unclear whether somatic mutations are a cause of functional decline in human health, for instance, aging, although the hypothesis was proposed in the 1950s, due to several critical knowledge gaps and important challenges. Due to cell-to-cell heterogeneity, somatic mutations are unique in each cell of normal tissues and are extremely difficult to detect accurately, which leads to the large unknown about somatic mutation burdens per cell with respect to tissue types and life span in normal, non-cancer tissues of an organism. In addition, the causes of somatic mutations and how they evolve in normal tissues during different life stages remain unclear. With the experience and expertise in somatic mutations and genome instability, our goals for the next five years is to utilize cutting-edge DNA sequencing technologies, including single-cell whole-genome sequencing, Nanorate sequencing, and ultra-deep bulk whole-genome sequencing and computational analyses to 1) quantitatively explore somatic mutation burdens in normal, non-cancerous tissues in mice across different life stages; 2) identify mutational signatures and clusters, to infer molecular mechanisms and risk factors and to guide experimental validation in the future; 3) decipher the patterns of somatic mutation evolution, examining whether they are subject to random drift, positive selection or negative selection across tissue types and life stages. Based on the new discoveries, for the long-term goals, I will explore the health relevance of mutation burden, signature, and somatic evolution, quantify the risks of environmental factors, and develop new standards for drug safety. Additionally, I plan to select specific genes or genetic regions (with either higher or lower mutation burdens than expected) for testing their health relevance. These goals will be achieved using mouse as the model organism for testing and developing new interventions. Overall, the proposed projects will provide comprehensive insights into somatic mosaicism, laying the groundwork for understanding its role and the mechanisms of its impact on biology and health and shedding light on future interventions.

NIH Research Projects · FY 2026 · 2025-08

Project Summary Epstein-Barr virus (EBV) remains a significant burden to global health. This orally transmitted virus is associated with a spectrum of diseases, including nasopharyngeal carcinoma, a type of head and neck cancer, oral hairy leukoplakia, infectious mononucleosis, and certain B cell malignancies, and about 10% of gastric cancers. Despite its substantial health impact, there are currently no commercial vaccines or targeted therapies for EBV infection or EBV-associated diseases, and the mechanisms underlying EBV's diverse pathogenic effects remain largely elusive. A critical aspect of EBV's oncogenic potential lies in its ability to transform primary B cells into immortalized lymphoblastoid cell lines (LCLs), a process central to EBV-associated tumorigenesis. EBNA2, an EBV encoded nuclear antigen, plays a pivotal role in this transformation by acting as a master regulator of both viral and host gene expression. EBNA2's pathogenesis is intrinsically linked to its transcriptional regulation activity. This protein regulates the expression of hundreds of viral and host genes. However, the mechanisms through which EBNA2 regulates gene expression remain elusive. In this proposal, we aim to elucidate the mechanisms by which EBNA2 regulates gene expression and its contribution to B cell transformation and EBV pathogenesis. We will investigate this from three key aspects: (1) Identification of EBNA2 co-factors involved in EBNA2- mediated transcriptional regulation. EBNA2 regulates gene expression through largely unidentified co-factors. Dissecting these crucial co-factors will not only enhance our understanding of EBNA2's regulatory mechanisms but also identify potential targets for treating EBV-associated diseases. (2) Determine the mechanism through which EBNA2 regulates genome organization and its impact on B cell transformation and gene expression. Our preliminary data indicate that EBNA2 globally alters host genome structure. Given the critical role of genome structure in gene transcription regulation, we hypothesize that EBNA2 manipulates genome organization to regulate gene expression. We will investigate the mechanisms through which EBNA2 regulates genome organization and its effect on gene expression and B cell transformation. (3) Single-cell analysis of EBNA2 in transcriptional regulation. LCLs are heterogeneous cell populations. Consequently, studying the EBNA2 regulome using bulk RNA-seq from LCLs may obscure crucial information about differential responses to EBNA2 among distinct cell subpopulations. To gain a more comprehensive understanding of EBNA2's impact on transcriptional regulation across diverse cell populations in LCLs and during EBV-mediated B cell transformation, we will employ single-cell RNA-seq and ATAC-seq techniques. In summary, this proposal will comprehensively evaluate the role of EBNA2 in transcriptional regulation. This research addresses critical knowledge gaps in EBV biology, potentially leading to new targeted therapies and preventive strategies against EBV-associated malignancies.

NIH Research Projects · FY 2025 · 2025-08

Persistent differences in cancer incidence, mortality, and survival across population subgroups cannot be explained fully by traditional risk factors. These differences are seen for all cancers combined, most common cancers, frailty, morbidity, and mortality among cancer survivors. Complex interactions of biologic, psychosocial, socioeconomic, and environmental factors contribute to these differences. This project, responsive to PAR-23-255, will use data from two well-characterized prospective studies, the Atherosclerosis Risk in Communities (ARIC) Study and Multi-Ethnic Study of Atherosclerosis (MESA), to investigate contributions of person-level and neighborhood-level social and structural determinants of health (SSDOH) to accelerated biological aging, cancer risk, and differences in frailty and mortality among cancer survivors. We will examine independent and joint contributions of education, economic stability, social isolation and support, neighborhood demographics and area deprivation ascertained at the census tract level, to study outcomes. This proposed study builds on our existing work (R01CA267977) that has used published and novel proteomic aging clocks (PACs) to quantify biologic aging and link PAC acceleration to total cancer risk, risk of smoking-related cancers, and frailty and mortality in cancer survivors in ARIC and MESA. With this R21, we seek to better understand the SSDOH drivers of age acceleration and their contribution to cancer risk and survivorship. By examining person-level and neighborhood-level SSDOH measures, we aim to address the complexity of variations in cancer incidence and survivorship. We have SSDOH, proteomic and cancer data on >10,800 ARIC participants with nearly 4,100 incident cancer cases over 30 years of follow-up, and >5,300 MESA participants with nearly 700 incident cancer cases over 18 years of follow-up. Our central hypothesis is that SSDOH become biologically embodied via PAC acceleration leading to disparities in cancer risk and survivorship outcomes. Specific Aims are: 1) Identify associations of person-level and neighborhood-level SSDOH with PAC acceleration and incident cancer in two prospective cohorts of adults. 2) Determine associations of both person-level and neighborhood-level SSDOH with PAC acceleration, frailty, and all-cause mortality among cancer survivors in ARIC and MESA. Our proposed analyses will be completed in the total population of ARIC and MESA (separately) and then stratified by population subgroups; subsequently we will complete meta-analyses combining ARIC and MESA data. Our multi-disciplinary study team is well-prepared to lead this work, with complementary expertise in research on population health differences, SSDOH and chronic disease risk, molecular and cancer epidemiology, oncology and cancer survivorship, aging biomarkers, biostatistics, and proteomics analyses. The proposed study will have a significant impact by advancing the understanding of how socio-environmental factors translate into biological age acceleration, measured by PACs, and lead to greater cancer risk and adverse outcomes among cancer survivors.

NIH Research Projects · FY 2025 · 2025-08

Abstract Raised intracranial pressure (ICP) is a common complication of cryptococcal meningitis (CM), affecting around half of patients, leading to increased morbidity and mortality. Lumbar punctures (LP) are used to diagnose raised ICP, by measuring cerebrospinal fluid (CSF) opening pressure (OP), and to treat raised ICP by therapeutic drainage of CSF. This reduces mortality but relief is often only temporary and high ICP frequently recurs during treatment, requiring further repeat LPs, sometimes daily. LPs are invasive, often painful procedures, and can lead to complications. A non-invasive test to identify patients with raised ICP would be highly advantageous as it would help identify patients at risk of ICP-associated complications, and significantly reduce the number of LPs performed purely for ICP monitoring reasons. Retinal imaging technologies show great promise and are now routinely used to detect and monitor raised ICP in other neurological conditions. These include Optical Coherence Tomography (OCT), which uses safe low- power laser to produce high-resolution, cross-sectional images of the retina, allowing for very accurate measurement of optic nerve swelling - a strong predictor of raised ICP; and fundoscopy, to identify retinal vessel changes associated with raised ICP. With the development of low-cost, portable OCT machines and lens adaptations that enable the use of mobile platforms for high-quality non-mydriatic fundoscopy, these assessments are now much more accessible to non-ophthalmologists and in resource limited settings. In this proof-of-concept translational proposal, we will evaluate whether mobile OCT and smartphone fundoscopy can be used to identify high ICP in CM patients at diagnosis and during treatment, and which non- invasive retinal imaging modality is better. To do this we will conduct a prospective cohort study in Kampala, Uganda, enrolling patients with HIV-associated CM. We will perform serial OCT and smartphone fundoscopy during inpatient treatment and compare these readings to opening pressure measured at LP. The proposal draws together expertise in retinal imaging and OCT analysis from Dr. Mollan’s group in Birmingham, with the clinical experience of a long-established and highly productive meningitis research collaboration between the University of Minnesota and the Infectious Diseases Institute, Makerere University (Uganda). This project has the potential to significantly impact the complex clinical care required to treat persons with HIV-associated cryptococcal meningitis, particularly in resource-limited settings. Using mobile retinal imaging as a non-invasive test for raised ICP would reduce the material resources, technical manpower, and possibility of adverse events that are associated with performing LPs. By demonstrating proof-of-principle and technical feasibility, the data from this R21 could then be leveraged into a larger diagnostic package of care used in a R01-funded randomized control trial for novel treatments and management strategies to reduce ICP in CM.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT A remarkable change in the approach to the diagnosis, evaluation, and management of congenital cytomegalovirus (cCMV) infection has occurred in recent years. Driven by the advent of universal newborn screening for cCMV, newborns with this infection that would have never been clinically recognized in past years are now being identified, evaluated, and monitored – primarily for delayed-onset sensorineural hearing loss (SNHL). SNHL is the major known complication of cCMV, occurring in up to 12% of infants with this infection. However, the advent of universal screening has created challenges, and exposed significant knowledge deficits about the natural history of cCMV. Conventional thinking has been that the majority of cCMV infections are asymptomatic (AcCMV), and except for those children that develop SNHL, the prognosis for a normal outcome is favorable. This assumption is based on small, often uncontrolled studies. The long-term neurodevelopmental and neurocognitive outcomes of AcCMV have never been examined in a well-powered and unselected population of infants diagnosed in the context of universal screening. To address these knowledge deficits, and better understand the potential range of outcomes in AcCMV, this proposal aims to address three specific aims. First, we aim to determine if infants with AcCMV have reduced white matter structural coherence when compared to control infants. Our control infants will come from the NIH-funded HEALthy Brain and Child Development (HBCD) Study, which is advancing the field of cognitive neuroimaging by developing and validating significantly faster and higher resolution acquisition protocols, resulting in more robust infant imaging data. Second, we will determine if infants with AcCMV have reduced performance on longitudinal neurocognitive assessments when compared to HBCD control infants. Key features will be standardized scores of cognitive, language, and motor skills, as well as executive function and emotion regulation skills. Third, we aim to explore new scientific avenues by determining if AcCMV-associated SNHL can be predicted with brain MRI. Multi-shell diffusion imaging data will measure axonal microarchitecture and fMRI passive listening tasks will measure task-specific BOLD responses. This aim addresses a major challenge in the field of cCMV, and the public health and clinical management implications are extensive. Our over-arching hypothesis is that infants with AcCMV do, indeed, have a long-term phenotype that has heretofore been unrecognized because of the limited sensitivity of previous approaches. This work will inform and direct the future of newborn cCMV screening programs, and has implications for clinical management, including antiviral therapy. The proposed project will also enhance our understanding of this ubiquitous but understudied congenital viral infection.

NIH Research Projects · FY 2025 · 2025-08

Processing complex sensory information requires highly interconnected cortical networks. A critical feature determining the behavior of these networks is the structure of lateral interactions between nearby neurons, which can, under certain circumstances, give rise to organized, modular functional activity. However, despite the clear importance of lateral interactions in a wide range of network models, we lack a clear understanding of their organization in vivo. To address this challenge, we propose a tight integration between computational modeling and optogenetic experiments at mesoscopic and cellular scales. We focus on a broad class of models explaining the emergence of highly organized modular patterns through structured local lateral interactions that exhibit local excitation—lateral inhibition (LELI) structure. The behavior of networks governed by such interactions depends critically on the spatial extent, heterogeneity, and net strength of these interactions, yet these essential parameters have not yet been examined in vivo. Here, we propose to directly probe local lateral interactions of cortical circuits, determine how these circuits transform local and large-scale inputs into output activity patterns, and develop computational models describing this transform to estimate these essential parameters of network interactions. By mapping functional interactions at mesoscopic and cellular resolution in vivo in a species with modular organization similar to that seen in humans, we will be able to provide the clearest evidence to date regarding the LELI- structure of lateral interactions. By applying specifically designed mesoscopic optogenetic stimuli in conjunction with widefield calcium imaging, we will map the transformation of input within the cortical network on millimeter scales, and shed light on the underlying lateral interactions. Further, by applying this framework to a brain area beyond visual cortex, our work has the potential to identify universal principles governing cortical development and function. The new insights from this proposed work will have broad implications, and will contribute to a deeper understanding of the circuit mechanisms responsible for the generation of cortical network activity states, providing fundamental knowledge that could serve as the basis for the development of next generation prosthetics and novel treatments for a host of neurological disorders. RELEVANCE (See instructions): The research in this proposal will provide new insights into how circuits within a nonmurine cortex transform incoming information. We will gain insights into how network interactions and their transformation of information may be perturbed in neurological disease. Ultimately, understanding the mechanisms underlying activity transformation within the cortex may provide avenues for potential therapeutic interventions in these diseases.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDA) is a lethal disease notoriously resistant to therapy including immune checkpoint blockade. Meanwhile, immunotherapy targeting the PD-1:PD-L1 pathway is inducing stunning clinical outcomes in other advanced malignancies. However, the underlying mechanisms governing immune surveillance and resistance to a robust antitumor T cell response in pancreatic cancer are largely unknown. We are poised to identify how to safely promote T cell-mediated destruction of pancreatic cancer through 2 funded R01 applications and a collaborative P01 application. Our preliminary data support the hypothesis that immune- mediated pancreatic cancer eradication requires the following components: 1) a high affinity tumor specific T cell, 2) modification of suppressive intratumoral myeloid cells, and 3) overcoming chronic inflammatory and immunosuppressive signaling pathways. Notably our results demonstrate that developing combinatorial strategies to overcome endogenous T cell dysfunction in PDA may not be the same for enhancing cellular therapy using T cell receptor (TCR) engineered based approaches. Thus, the 3 funded projects are based on the development of highly faithful PDA animal models in which an endogenous or TCR engineered T cell response can be tracked longitudinally over time. The projects are aimed at uncovering novel mechanisms that interfere with the efficacy of combinatorial immunotherapies designed to engage the endogenous immunity (project 1) or TCR engineered T cell therapy (project 2 and 3). We will test novel combinatorial approaches to safely enhance the antitumor activity of a combination therapy that uses CD40 agonist, anti-PD-L1 (project 1) or abrogating TGFb signaling in TCR engineered T cells (Project 2) or the combination of TCR engineered T cell therapy and simultaneous modification of the desmoplastic tumor microenvironment (Project 3). Together the studies will identify characteristics of T cells and the tumor microenvironment that produce durable antitumor responses during immunotherapy.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Understanding how neuronal circuits give rise to behavior requires recording their activity at a high temporal and spatial resolution with cell type specificity. This is particularly critical in the case of the cerebellum, which in addition to being essential for effective control of movement, has ever emerging roles in cognitive functions and neurological disorders. However, such recordings have historically been challenging, due to the complex and conserved cytoarchitecture of the cerebellar cortex, and the firing properties of its primary output neurons, Purkinje cells, which exhibit two distinct activity modalities: simple spikes and complex spikes. Optical interrogation of Purkinje cell Ca2+ can report complex spike events due to their low frequency (0.5-2Hz), but the slow kinetics of Ca2+ indicators make them unviable for resolving simple spikes. Conversely, while microelectrode arrays offer high temporal resolution, they provide limited spatial information. Here, we propose to leverage ultrasensitive genetically encoded voltage indicators (GEVIs) in order to perform the first optical voltage recordings of Purkinje cell simple and complex spike activity during behavior. Our approach will allow us to examine the encoding properties of Purkinje cell simple and complex spiking modalities during behavior and interrogate their interactions relative to the geometry of their inputs and overarching functional organization. In Aim 1 of this proposal, we will establish the instrumentation and procedures for ultra-high-speed voltage imaging of cerebellar Purkinje cells in awake behaving mice. Using this approach, we will examine spike level dynamics of both simple and complex spike firing, computing spatiotemporal correlation and activation patterns of neighboring Purkinje cells. We will also interrogate the kinematic encoding properties of these two activity modalities. In Aim 2, we will combine our approach for voltage imaging with simultaneous Calcium imaging of dendritic activity, to compare Purkinje cell encoding properties and responses to sensorimotor mismatches, whose processing is considered a hallmark of cerebellar function, with their overarching functional dendritic organization. Together, our work will shed new light on the population dynamics of Purkinje cell spiking activity and contributions to behavior, as well as provide new potential links between the functional and structural organization of the cerebellar cortex.

NIH Research Projects · FY 2025 · 2025-08

Tuberculosis (TB) is the leading cause of death by a bacterial disease worldwide. Differences in lung microenvironments, including the granuloma, influence the support and suppression of Mycobacterium tuberculosis (Mtb). While factors that impact microenvironment specific immune responses and Mtb sterility in the lung are incompletely understood, recent studies have shown that the microbiome takes an active role in shaping the immune response. Both lung and gut microbiomes have been shown to influence disease outcome in influenza, cystic fibrosis, asthma, cancer, and COPD. The dominant animal model used in TB is the mouse. While laboratory breeding of mice in specific pathogen free (SPF) and hygienic conditions has increased reproducibility, it has also removed much of the microbial exposure experienced by free-living organisms. Daily encounters with microbes help sculpt the immune system for a robust response against new pathogens. Unfortunately, the immune system of SPF mice lacks effector differentiated CD8+ memory T cells, which are critical cell types for an immediate response to infection. It is unknown how prior experience to microbes impacts TB progression. This proposal leverages a novel mouse model (‘dirty’ mouse model) that creates multiple exposures to diverse microbes to shape the immune response in SPF mice. Microbially experienced pet store mice will be housed with C3HeB/FeJ SPF mice; thereby, transferring both commensal and pathogenic microbes between animals. C3HeB/FeJ mouse strain is selected as it forms multiple types of TB granulomas similar to human TB. The overall goal of this project is to determine how prior diverse microbial experiences influence the course of TB infection (‘dirty’ vs. SPF mice). The central hypothesis is that the lung and gut microbiomes of dirty C3HeB/FeJ mice will maintain equilibrium in the face of infection and shape the immune system to resist and control Mtb as compared to SPF mice. This proposal will define changes to 1) lung and gut microbiota diversity, 2) locations and interactions of microbiota inside granulomas and lungs, 3) innate and adaptive immune cell populations, and 4) IL-23 pathway, a critical host response in the control of TB, in dirty and SPF mice. The dirty mouse model will serve as a complement to existing small animal models of TB to reveal new biomarkers and host-directed treatments to TB that are influenced upon prior microbial experience. The outlined studies push the boundaries of our current understanding of TB pathogenesis and may ultimately show TB as a disease that is defined by intimate interactions between the pathogen, host, and microbiota.

NIH Research Projects · FY 2025 · 2025-08

Project summary This proposal seeks the acquisition of a LUMICKS C-Trap instrument at the University of Minnesota. The C-Trap is the only commercially available laser optical trap that combines TIRF fluorescence microscopy with integrated microfluidics in a user-friendly, reliable, engineered instrument. Single molecule force and fluorescence spectroscopies have transformed the fields of cellular and molecular biology, biochemistry, biophysics, and biomedical engineering, providing answers to crucial biological and health-related questions. These tools are currently limited to laboratories with specialized expertise and custom-built single molecule instruments. The C-Trap is designed as a turnkey system, enabling laboratories without specific single molecule force or fluorescence spectroscopy expertise to conduct advanced single molecule force, position, and fluorescence localization, as well as FRET analysis, with unparalleled spatial and temporal resolution. Key features of the C-Trap include dual optical traps for manipulating biomolecules with sub-pN force resolution/detection; 3-color laser TIRF and widefield fluorescence detection for visualizing biological processes; precise temperature control; micro and nano-stage control for exact sample positioning; laminar flow microfluidics for high sample throughput and varying ambient media while maintaining molecular interactions; and user-friendly software for integrated, controlled, and automated instrument operation. The intuitive instrument and software interface, along with the automation package, will enable non-expert users to run experiments and collect high-quality data with minimal training. The proposal represents five major users (all with NIH R01/R35 funding) who will primarily use the instrument, and eight minor users with NIH or other funding. These users come from four colleges across University of Minnesota (College of Biological Sciences, College of Science and Engineering, School of Dentistry, and the Medical School) and six different departments. The C-Trap will significantly advance the NIH-funded projects of these researchers by enabling high-impact single molecule mechanobiological and fluorescence studies on a range of important biomedical questions in areas such as cell surface mechanosensing, muscular dystrophy pathogenesis, cytoskeletal structure and dynamics, viral assembly and cell entry and nucleic acid processing. The major users and technical advisors possess extensive relevant expertise with the technology, and will provide guidance to all users. The instrument will be housed in the University Imaging Center, a Nikon Center of Excellence, which provides support and training for all of their instruments as well as expert advice and consultation, from experiment planning to image acquisition and analysis. Establishing this resource will significantly enhance numerous research programs at the University of Minnesota and contribute to the discovery of impactful scientific, biomedical, and healthcare-related knowledge.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Predicting Late Talkers in Infants who are at Elevated Familial Likelihood for Autism Autistic children are late to say their first word, and these early challenges persist with over 70% of autistic preschoolers having language impairment. Siblings of autistic children unaffected by autism themselves are at a 4-5-fold increased risk of developing language challenges. The field lacks screening tools with strong predictive power to identify late talking autistic children during the birth-to-three early intervention period. The proposed project makes significant steps towards early identification of language impairment and understanding the developmental sequelae of language in autistic toddlers by leveraging data from the Baby Sibling Research Consortium database, the largest collection of language data of infants who develop autism. Aim 1 of the proposed study is create a normative model sensitive to the heterogeneity of autistic language development using summary-level data from the MacArthur-Bates Communicative Development Inventories (CDI). This model will then be used to predict late talkers. The normative modeling framework quantifies individual differences in scores, flagging individuals for further follow up. This framework moves away from the limitations of simple “case- control” designs by allowing for the heterogeneity that is inherent to developmental disorders. We will make our normative model publicly available, to be used by investigators interested in samples that tend to be smaller in size, increasing the significance and