University Of Minnesota

NIH Research Projects · FY 2025 · 2018-08

PROJECT SUMMARY / ABSTRACT Listening effort has large impact on quality of life, and thus warrants detailed and thorough study. There has long been a conundrum over what effort is and how to measure it. We propose three new branches of work that follow in the spirit of our previous grant cycle, building off of the key observations that listening effort is tightly related to comprehension rather than phonetic accuracy, and that effort can emerge at distinct moments in time before, during and after the listening process. The new aims focus on the effort of preparing to listen, the effort of aiming attention at particular moments during listening, and the effort that lingers while a person is comprehending the meaning of what was heard. The experiments are guided by the lived experiences of people with cochlear implants, who regularly offer interpretation and suggestions on studies that illuminate the situations that pose recurring problems in their daily communication. Our team of clinically trained scientists and language/hearing experts integrates this input into rigorous designs that capitalize on our strength in creating stimuli, measuring listening effort, and sharing our resources with the field. Different aspects of effort need to be distinguished in order to better alleviate the associated costs. For example, effort of recovering from mistakes is different than the anxiety of listening. Overall high overexertion could be distinguished from exertion that simply lasts too long. The proposed work will build understanding of how effort changes on a moment-to-moment basis before, during, and after listening, and how this is affected by hearing difficulty – particularly the use of a cochlear implant.

NIH Research Projects · FY 2026 · 2018-08

SUMMARY Evaluating the impact of a potential tobacco product standard requires a comprehensive understanding of the direct impact of the intervention, as well as an understanding of the unintended consequences and other down-stream effects to capture the full effect of the regulation on public health. To support FDA regulatory action, impact analyses must be supported by rigorously designed studies and innovative statistical methods. We propose to develop novel statistical methods for evaluating the impact of a tobacco product standard. We motivate our development by gaps in the literature related to nicotine reduction, but many of the barriers to evaluating the impact of nicotine reduction are challenges for other tobacco product standards, as well. Since 2010, the FDACTP and NIH have funded many randomized controlled trials (RCTs) to understand the impact of a low nicotine standard for cigarettes, yet, critical unanswered questions remain related to 1) the impact on cessation, 2) understanding the population-level impact calibrated to the United States smoking population, 3) understanding the heterogeneity in the effect of a low nicotine standard to achieve an equitable impact across the population, and 4) estimating the real-world impact of nicotine reduction. Addressing these questions is challenging due to no single data source providing sufficient power or precision (i.e. cessation), lack of a sufficiently diverse and representative population (i.e. calibration and heterogeneity), or fundamental differences between implementing the intervention in practice and in a RCT (i.e. real-world impact). To understand the full impact of potential regulatory action, we must integrate information from multiple data sources, but existing approaches to data integration are insufficient. For the last seven years, we have developed statistical methods for tobacco regulatory science (TRS), providing the statistical tools needed to answer the questions most relevant to FDACTP. In this application, we address limitations in the existing methods for data integration that prevent the TRS community from answering key questions through the following specific aims: 1) Develop causal meta-analytic techniques for estimating the regulatory effect of an intervention; 2) Develop a Bayesian approach to dynamic borrowing that leverages the underlying latent structure of the data; 3) Develop methods to estimate the individual treatment effects to understand disparities in the impact of regulations; 4) Develop approaches to sensitivity analysis to understand the effect of unmeasured factors on the estimated regulatory effect of tobacco product regulation. This application addresses FDACTP scientific interest “Impact Analysis – Understanding the impact of potential FDA regulatory actions”. Our proposal represents a significant contribution to TRS and will result in statistical methods that provide more precise estimates of the impact of FDA regulatory action. The innovative methods proposed in this application will provide powerful tools for leveraging the extensive data resources for evaluating the impact of tobacco product standards.

NIH Research Projects · FY 2025 · 2018-07

Project Summary This proposal is for a competitive renewal of the NS105604 training grant under the Jointly Sponsored NIH Predoctoral Training Program in the Neurosciences (JSPTPN) at the University of Minnesota. The program has been very successful over the last five years, setting 32 predoctoral trainees on the start of their graduate career. It continues the University of Minnesota’s highly successful NIGMS training program, “Predoctoral Training of Neuroscientists” (T32-GM08471,1993-2018) under the Systems and Integrative Biology Program. Trainees in this program are pursuing a PhD through the Graduate Program in Neuroscience at the University of Minnesota, an interdepartmental and interdisciplinary program that spans 30 departments. This proposed program provides select trainees added value in the first two years of their graduate careers, providing its trainees with a broad foundation in neuroscience as well as the interdisciplinary skills needed to be successful within their neuroscientific careers. In particular, the program is designed to provide a broad understanding of the field as well as a deep understanding of research methodologies, experimental design, and quantitative reasoning. This program is built around a core of didactic coursework in neuroscience, exposure to research-related issues such as ensuring rigor and reproducibility and quantitative analyses, and the beginning of thesis-related in-depth research projects. Flexibility and time for in-depth development of collateral fields of knowledge are provided. Several unique educational opportunities are included, including the long-running Itasca summer laboratory, rotations, and specific classes in rigor, reproducibility, and quantitative reasoning. Time and attention are given to the student’s professional development, including exposure to neuroscience at the national and international levels. A talented group of trainers have been assembled that reflects the diversity of research questions, areas of study, and techniques in neuroscience. Each trainer directs a productive research program and has demonstrated commitment to teaching and training. The trainers are united by their participation in the Graduate Program in Neuroscience and by their dedication to predoctoral training. An impressive array of scientific and institutional resources are available to the trainees, including substantial direct institutional support for this training grant. The graduates of this program will be trained to be independent researchers, capable of making contributions in academia, teaching, industry, government, and public service.

NIH Research Projects · FY 2026 · 2018-05

The Gardner Laboratory uses a combination experimental and computational approach to dissect molecular mechanisms for how microtubule lengths are regulated inside of cells, and for how forces within the mitotic spindle act to ensure proper chromosome segregation during mitosis. We use biophysical computational modeling to better integrate and understand our experimental observations, make new experimental predictions, and to test whether our proposed cellular mechanisms are physically reasonable. Overall, we are a cellular biophysics laboratory that combines cell biology tools with biophysical methods to shed new light on the regulation of microtubule dynamics, and to dissect forces and their consequences in mitosis. Achieving the goals described in this application will provide mechanistic insights into how molecular-scale changes in microtubule structure could regulate cellular-scale changes in microtubule-associated protein localization and binding, as well as how controlled mitotic forces could act to protect genome integrity during mitosis. In particular, this application will advance our understanding of: 1) how microtubule structure can alter protein binding, and vice versa, 2) how the cell reads out and responds to nuanced tension signaling during mitosis, and 3) how controlled anaphase forces could act to protect genome integrity during anaphase.

NIH Research Projects · FY 2026 · 2018-05

Project Summary Cell signaling and membrane traffic emerge from an ensemble of dynamic, transient protein-protein interactions (PPIs) in a crowded milieu. Traditional structural and biochemical approaches are mostly limited to dissecting the function of stable, structured PPIs. To address emergent function stemming from transient PPIs, my research program develops innovative protein engineering and biophysical technologies. We investigate outstanding questions in GPCR-G protein selectivity and cell surface receptor activation of myosins. Studies will advance the fundamental cell biology of GPCRs and myosins, while delivering new therapeutic strategies to combat disease. Building on new technologies and conceptual advances from my lab, we propose five parallel research projects. (1) We discovered and characterized the temporal coupling of sequential GPCR-G protein interactions, leading to allokairic modulation of GPCR signaling. We will dissect the structural basis of allokairic modulation through the GPCR’s sequence-divergent third intracellular loop (ICL3). Using novel biosensors and receptor chimeras, we will define roles for ICL3 in autoregulation and G protein selection in closely related receptor isoforms. (2) We engineered a simple, accessible cell-free biosensor assay to measure the molecular efficacy of GPCR ligands. We will use this assay to identify and characterize receptor isoform-selective biologics, including peptides, peptide-mimetics, and nanobodies/affibodies. These biologics will serve as probes to advance the structural basis of GPCR-G protein selectivity and yield cell-permeable strategies to selectively target GPCRs. (3) We successfully integrated a computation-experiment collaboration to reveal the dynamic reshaping of GPCR cytosolic cavities underlying G protein selection. Using this strategy, we will map temporally persistent receptor- G protein interaction hot-spots across GPCRs, that encode G protein selectivity. We will dissect the structural basis of allosteric modulators through the dispersal of inter-residue communication networks within GPCRs. (4) We identified motor-cargo interaction kinetics and mechanical stiffness as two novel cellular regulatory mechanisms of cytoskeletal motors. We will use programmable biomimetic scaffolds to dissect myosin regulation through both receptor-adaptor and adaptor-motor ensembles. We focus on the impact of motor conformation and clustering triggered by diverse cell surface receptors including β1-integrin, plexin D1, and LRP2/megalin. (5) We will investigate a novel temporal bias mechanism in GPCR signaling, through receptor-mediated engagement of myosins during membrane traffic. We will characterize the differential regulation of motor activity through PDZ-binding motifs in the GPCR C-tail. We will use optogenetic/chemogenetic strategies to steer GPCR trafficking and map the temporal signaling profile through second messenger and Akt/MAPK pathways.

NIH Research Projects · FY 2026 · 2018-04

ABSTRACT Chronic kidney disease is a significant healthcare issue affecting >15% of the U.S. population and costing billions in healthcare dollars annually. Transplantation is the best option for most patients with progressive disease, resulting in a significant increase in life expectancy and improved quality of life compared to dialysis. The potential U.S. deceased donor organ supply is estimated to exceed the current number of organs transplanted by a factor of 4- to 5-fold, with a major limitation to the number of acceptable organs for transplant being the ischemic injury sustained between recovery and implantation. A method to cryopreserve or “bank” kidneys prior to transplant would effectively remove the influence of time from the supply chain of organ distribution. This would allow a new paradigm for transplantation that would improve donor/recipient matching, allow for better patient preparation, facilitate tolerance induction protocols, and increase organ utilization while improving graft and patient survival. One promising approach that overcomes the limitations of conventional strategies is vitrification—that is, cooling organs so quickly that they cannot undergo the phase transition from liquid to solid ice. With the help of cryoprotective agents (CPAs), the organ enters a stable glass-like state wherein viable storage is theoretically indefinite. The critical challenge, however, is rewarming without ice formation or cracking: if rewarming is too slow, ice crystals form, and if rewarming is not uniform, thermal stress causes cracking. During our initial R01 funding, we developed a novel approach termed “nanowarming” that achieved both objectives. Iron oxide nanoparticles were perfused throughout the vasculature of the organ along with CPA solutions. The organ was then vitrified by cooling and rewarmed on-demand by placing it in a radiofrequency coil that induces heating in the nanoparticles and, therefore, from within the organ. We found that nanowarming could rewarm vitrified organs, including kidneys, in animal models. We have recently shown, for the first time, that nanowarmed organs function in vitro and in vivo following transplantation. Further, we showed successful vitrification and nanowarming of human-sized (porcine) kidneys. These new data support the feasibility of our approach to cryopreserve and nanowarm whole human organs for transplantation. Nevertheless, many questions remain, including how nanowarmed kidneys function compared to control organs, what, if any, injury occurs during nanowarming, and how to scale up to human-sized organs. In this renewal R01, we propose to: (1) Quantitatively assess cryopreserved and nanowarmed kidney transplant function in a rat model, including long- term preservation, long-term function, modes of injury, and alterations of the host immune response, (2) Engineer and optimize scale-up for nanowarming vitrified human-sized organs, and (3) Vitrify and nanowarm human-sized kidneys while measuring viability, structural integrity, and organ function.

NIH Research Projects · FY 2025 · 2018-04

PROJECT SUMMARY/ABSTRACT Synthesis, Structure, and Mechanism of Biorelevant Molecules and Reactions Our MIRA-supported research program encompasses synthetic, mechanistic, and structural organic chem- istry. We address unresolved contemporary problems through studies that lead to i) new ways of deducing the structures of novel chemical entities, often through innovative use of NMR methodologies, ii) new insights about how chemical reactions, including spontaneous biosynthetic transformations, proceed, and iii) new ways to make molecules that have structural features of interest to researchers pursuing targets with promising bio- logical properties. We will capitalize on recent accomplishments and launch new efforts as follows. I. Natural Products Chemistry A. We remain interested in unraveling key steps in the biosynthesis of natural products that proceed in the absence of enzymatic catalysis—that is, spontaneously. Two specific hypotheses related to the origin of the unique skeleton of ottelione A drive current work: i) an unprecedented, low-barrier Cope rearrangement fashions the strange, dearomatized 4-methylenecyclohexenone present in this secondary metabolite and ii) a simple, achiral diarylheptanoid is oxidatively transformed into the strained and preorganized Cope substrate. The engagement of an outstanding collaborator to use genome mapping approaches will be of great benefit. B. We frequently engage in natural product structure determination studies and the development of methodologies of value to those who do the same. These studies have had impact extending well beyond the specific questions that we address. Our record in doing this is strong. One notable example teaches methodol- ogy for calculation of chemical shifts to the experimentalist who may be a novice computationalist. Our ap- proach was the same as that used in the newest developments of probabilistic methods for comparison of computed vs. experimental NMR chemical shifts to validate structure assignments (DP4, DP4+, DP4-AI, DP5). We find a gap in that some communities have yet to embrace these approaches. We propose to evaluate the effectiveness (and limitations) of these methods for structural assessment of various cyclic peptides and then to communicate, advertise if you will, these outcomes to benefit future structural studies by peptide chemists. II. HDDA-Benzyne Chemistry Our discovery of the broad scope of the hexadehydro-Diels–Alder (HDDA) reaction is both exciting and en- abling. This work has advanced significantly since the onset of our MIRA funding four years ago. The opportu- nities in this arena show no sign of abating. To the contrary, it seems that every month or so a coworker arrives at my doorstep with yet another new result that elicits from me something to the effect of “Wow, HDDA- benzynes will also do that!” Myriad new directions are presented in pages 4–6 of the Research Strategy. Many will lead to products containing a greater preponderance of heteroatoms, thereby demonstrating new ap- proaches for consideration and use by researchers engaged in drug discovery activities. (30 lines)

NIH Research Projects · FY 2026 · 2018-02

Project Summary Bruch’s membrane (BrM) represents a small portion of the human eye, yet its role in maintaining ocular health through regulating the transport of macromolecules and nutrients between the retinal pigmented epithelium (RPE), BrM, and choroid, is vastly larger. To highlight this fact, mutations in select proteins involved in forming one of the layers of BrM, the RPE basal lamina (BL), are linked to a spectrum of early- and late-onset macular diseases resembling age-related macular degeneration (AMD). For example, an autosomal dominant R345W mutation in fibulin-3 (F3, aka EFEMP1), a key RPE-BL protein, triggers Doyne Honeycomb Retinal Dystrophy/Malattia Leventinese (DHRD/ML), an aggressive macular dystrophy that closely resembles early/intermediate AMD, and can predispose DHRD/ML patients to advanced forms of AMD (geographic atrophy and neovascularization). We hypothesize that F3 is a key contributor to pathogenic sub-RPE deposits (basal laminar deposits [BLamD] in mice, sub-RPE basal deposits in RPE cultures, and drusen in humans). The long-term goal of our research effort is to develop tractable F3-centric therapeutics for DHRD/ML and early/intermediate AMD. To accomplish this goal, we will i) test small molecule-based anti-inflammatory strategies targeting BLamD formation, ii) test the effects of driving RPE-specific expression of F3 in the mouse RPE, and iii) identify RPE quality control pathways that regulate F3 proteostasis. Successful completion of this project will positively impact our knowledge of this important ECM protein and will provide additional tools and actionable information for interrogating/restoring the intricate biology underlying incurable inherited and age-related eye diseases.

NIH Research Projects · FY 2024 · 2017-09

Project Summary Migrating cells use filopodia to interact with and efficiently move through their complex 3D environments. Filopodia are slender actin-filled projections composed of a core of cross-linked, parallel actin bundles. They are highly dynamic, vary in length and found in a wide variety of cell types such as neurons that use them for gradient sensing and efficient directional migration or cancer cells that employ them for moving out from tumors into neighboring tissue. The first steps of filopodia formation are poorly understood. Three conserved proteins are required for their formation - a MyTH4-FERM myosin) and two regulators ofactin polymerization, VASP and Formin. How the action of these three proteins is coordinated to initiate filopodia formation is unknown. The objective of this proposal is to define the molecular mechanism of filopodia initiation with an emphasis on the role of a MF myosin and its functional relationship to the actin polymerase VASP in this process. Recent work revealed that activation and specific targeting of the MF myosin to the cortex requires the actin polymerization activity of the regulator VASP. The versatile model system Dictyostelium will be used to define the mechanism of this collaborative interaction. It will also be used to investigate how the myosin motor and actin regulator work together to organize the fast-growing ends of actin filaments at the membrane to initiate polymerization. A combination of in vivo, in vitro and in silico approaches will be employed to a) gain new insight into the regulation and mechanism of filopodia initiation and filopodial function in vivo; b) characterize the MF myosin motor and its interaction with the actin network in vitro; and c) build a predictive mathematical model of filopodia initiation. The knowledge generated by this project will reveal how cells use a myosin-based motor to build specific actin-based structures such as filopodia. Understanding how initiation occurs will also reveal how cells control filopodia formation to undergo directed migration or invade into surrounding tissues.

NIH Research Projects · FY 2024 · 2017-09

Project Summary/Abstract The mechanism of DNA packaging for double-stranded DNA viruses will be studied in the Bacillus subtilis bacteriophage f29, the most efficient in vitro viral packaging system known. Using an integrated genetic, biochemical, computational and structural approach, we will characterize protein conformational change and movement in the transiently assembled packaging motor during DNA encapsidation. The mechanism of packaging in f29 will serve as a model for animal virus packaging in the analogous herpesvirus and adenovirus systems, and aid in the search for new antiviral therapies. Due to similarities between the f29 ATPase and other ring translocases, insights gained from the study of f29 packaging will also provide insight into the basic principles of macromolecular motor function in higher organisms. To interrogate the mechanism of DNA packaging we will: 1. determine the molecular basis for force generation of the packaging motor; 2. elucidate the mechanism of nucleotide cycling for a complex multimeric ATPase; and 3. describe the nature of intermolecular communication that coordinates the action of a biological nanomotor.

NIH Research Projects · FY 2024 · 2017-09

PROJECT SUMMARY In this renewal, we seek to understand the origin of heterogeneity in sickle cell disease (SCD), which is present at every scale from molecules to the clinic, and is the major impediment to clinical management and the development of new therapies. Moreover, therapy often increases heterogeneity, with some patients responding strongly to therapy and others unresponsive. Our central hypothesis is that heterogeneity originates with intracellular kinetics of sickle hemoglobin (HbS) self-assembly that translates into heterogeneous populations of RBCs, which drive strong non-Newtonian fluid behavior in whole blood and alterations in the systemic circulation that precipitate pathologies such as endothelial injury, vaso-occlusion, aneurysm, and stroke. Thus, the ability to guide therapeutic intervention and to develop new therapies is ultimately hindered by our limited understanding of heterogeneity in the context of multiscale biophysical processes in SCD pathophysiology. In this work, we will develop a biophysical framework for SCD pathophysiology that spans from molecules to the systemic circulation, that is experimentally validated at every scale, and that allows us to predict the effects of multiscale heterogeneity. Specifically, we will: (1) Develop a quantitative framework for HbS polymerization that accurately predicts the kinetics of self-assembly; (2) Define the connection between the distribution of HbS polymer and mechanical properties among a population of RBCs; (3) Understand how cellular heterogeneity drives non-Newtonian blood rheology and altered flow in the systemic circulation. The work in this renewal builds on key conceptual advances made during our last 3 years of funding: HbS self- assembly kinetics have previously been underestimated by at least an order of magnitude; HbS polymer is heterogeneously distributed in RBCs at finite oxygen tension; velocity profiles in sickle blood demonstrate strong non-Newtonian effects; blood flow in SCD patients is altered throughout the circulation with aberrantly large wall shear stress relative to healthy blood. This work also leverages a unique and enabling set of tools that we have developed during the last 3 years of funding: the highest spatiotemporal resolution measurements of single HbS fiber assembly to-date; the first platform capable of quantifying HbS polymer in large populations of single RBCs under well-defined oxygen tension; a platform capable of quantifying viscoelastic properties of large populations of RBCs under well-defined oxygen tension; the ability to quantify submicron velocity fields in flowing blood at physiologic hematocrit; a platform to quantify sickle blood flow within physiologic oxygen gradients. Building on these tools and insights, this renewal work will develop and validate a multiscale model describing how heterogeneity propagates from the molecular to cellular to system levels, and we will develop experimental tools that can be used for clinical management and therapeutic development.

NIH Research Projects · FY 2026 · 2017-09

Project Summary Proline hydroxylation (Hyp) is a fundamental posttranslational modification and regulatory mechanism that are highly responsive to the changes in cellular metabolic conditions. During tumor progression, the rapid proliferation of cancer cells creates a hypoxic microenvironment that inhibits the hydroxyproline-mediated degradation of HIFa proteins and activates hypoxia-response cellular pathways to promote cancer cell survival in hypoxia. In addition to oxygen, the modification enzyme prolyl hydroxylases are also sensitive to the concentration of iron and key mitochondria metabolites including succinate, fumarate, and alpha-ketoglutarate, making the pathway a critical metabolic sensor in cells. Extensive studies have demonstrated that proline hydroxylation regulates protein structural stability, protein-protein interactions, or proteasomal degradation of substrate proteins. Despite its important roles in cell physiology and success in the targeted analysis of individual substrates, system-wide characterization and functional quantification of the pathway have been hindered by the lack of effective tools and strategies for site-specific identification of proline hydroxylation targets. Our overall hypothesis and long-term goal is that systematic characterization of “proline hydroxylome” through the development of functional proteomics approaches will lead to the mechanistic understanding of novel Hyp-mediated metabolic regulations in development and diseases. Moving towards this goal, in the past years, we have established HypDB for functional annotation analysis of the Hyp proteome with the development of a streamlined workflow for systematic analysis of the Hyp substrates in cells and tissues. We have gained extensive experience in biochemical characterization of Hyp targets, the interactome of specific prolyl hydroxylase as well as its crosstalk with other PTM regulatory pathways. To continue our effort, we will expand the HypDB to quantify Hyp dynamics in mouse tissues and develop functional proteomics strategies to identify key Hyp sites in protein structural stability and prolyl hydroxylase targets. We will apply recently developed chemical and biochemical strategies to investigate the crosstalk between proline hydroxylation and other metabolic-sensing modifications in regulating substrate protein degradation and activity. Furthermore, we will study the physiological significance of a new Hyp-mediated epigenetic modification pathway in regulating gene expression and chromatin activity. Overall, we anticipate that the development and application of functional proteomics technology for system-wide analysis of proline hydroxylation proteome will reveal novel metabolic-sensing pathways and potentially lead to paradigm-shifting concepts in the fields of cancer, metabolic diseases, and development.

NIH Research Projects · FY 2025 · 2017-08

Project Summary / Abstract Genetic variation among individuals shapes important phenotypes, including the risk for common human diseases such as cardiovascular, autoimmune, and neurological disease. In particular, regulatory genetic variation causes inter-individual differences in gene expression. The resulting gene expression differences account for a substantial portion of variation in many genetically complex traits. In spite of the critical importance of regulatory variation, many fundamental questions remain open. First, most DNA differences in a given genome likely have no effect. The nature of the specific variants that do have effects remains poorly understood. Second, genetic variation can specifically affect the protein abundance of a given gene without altering the abundance of the mRNA of the same gene. The mechanisms that are responsible for these protein-specific effects remain unclear. Third, we only have a crude understanding of how the differences in gene expression that result from regulatory variation affect organismal phenotypes. Over the next five years, research in my laboratory will focus on addressing these critical gaps in knowledge. Specifically, we seek to identify and characterize causal DNA variants, study the impact of genetic variation on protein degradation, and examine quantitatively how the precise abundance of a given gene can shape organismal traits. Our work combines computational biology, quantitative and statistical genetics with experimental genome-wide approaches. We use the yeast Saccharomyces cerevisiae as a powerful and tractable model system for regulatory variation, while pursuing related approaches in human cells. Our long-term vision is to improve our understanding of regulatory variation to the point at which it becomes possible to accurately predict the consequences of the DNA variants in an individual’s genome. This ability will be valuable for fundamental research and personalized approaches for improving human health.

NIH Research Projects · FY 2025 · 2017-08

Project Abstract: The University of Minnesota (UMN) Veterinary Diagnostic Laboratory (VDL) has the capacity and the opportunity to support the investigation of animal foodborne illness through the FDA CVM Vet-LIRN program, most specifically in the area of bacterial contamination.

NIH Research Projects · FY 2025 · 2017-08

PROJECT SUMMARY Although the USA has seen a significant increase in the number of PhD degrees in the biomedical sciences earned by scientists from underrepresented and racially minoritized (URM) backgrounds, there has not been a corresponding increase in the representation of URM faculty in the biomedical professoriate. To address this disparity in the state of Minnesota and more broadly, the University of Minnesota formulated and implemented the Minnesota IRACDA Program. The Minnesota IRACDA Program is a collaboration between the University of Minnesota and the nearby community colleges of Normandale Community College and North Hennepin Community College, which predominantly serve students from minority and low-income urban populations. The Minnesota IRACDA Program provides a diverse cohort of recent PhD scholars with rigorous scientific research training, instruction, mentoring, experience in teaching, and career development guidance. The long-term goal of the Minnesota IRACDA Program is to develop and implement successful and creative strategies to promote and facilitate the transition of talented postdoctoral researchers from diverse backgrounds into independent faculty careers in biomedical research and education. The overall objectives of the Minnesota IRACDA Program are to: (1) Recruit postdoctoral scholars from backgrounds that have been historically excluded from STEM research and education and provide them with strong multitiered mentoring in cutting-edge biomedical research. (2) Provide the postdoctoral scholars with pedagogical training in modern teaching techniques that incorporate diverse populations, and mentored practice in evidence-based teaching at the undergraduate level at two teaching-intensive institutions that predominantly serve students from minority and low-income urban populations—Normandale Community College and North Hennepin Community College. (3) Equip postdoctoral scholars with a multitude of career and professional development opportunities and programs including, participation in workshops that address scientific communication, publication, job searches, grant writing, professional advancement, and diversity, equity, and inclusion in biomedical research. (4) Provide URM undergraduates from the teaching-intensive partner institutions with paid authentic research opportunities at the University of Minnesota in which they are mentored by the postdoctoral scholars. In the first funding period, the Minnesota IRACDA Program met or exceeded all key benchmarks: (i) Nearly 40% of the IRACDA scholars recruited to the program were from URM backgrounds. (ii) IRACDA scholars were successful in career progression. Of the six scholars who completed training, four moved on to tenure-track Assistant Professor or Lecturer positions. (iii) URM undergraduate students participated in authentic research under the mentorship of IRACDA scholars and most successfully transferred to four-year degree programs. We are seeking funding to support six IRACDA postdoctoral scholars per year when the program is fully populated.

NIH Research Projects · FY 2026 · 2017-07

The ability to effectively study and develop innovative treatments for acute critically ill or injured patients remains challenging. Many of these devastating conditions occur infrequently while time-sensitivity of treatments challenge the capacity to obtain prospective informed consent, impeding the ability to enroll large numbers of patients into clinical trials. Yet, the current therapies for many of these devastating conditions are often understudied or unsatisfactory. To this end, the Strategies to Innovate Emergency Care Clinical Trials network (SIREN) provides a national structure to pool resources to address these challenges and advance emergency care. In this proposal we seek to continue our contributions to the SIREN network. These contributions include high numbers of high quality patient enrollments to all funded clinical trials, innovation and dissemination of pragmatic, cost-effective, and ethical advances in the conduct of emergency care research, support of prehospital and Exception From Informed Consent (EFIC) study procedures, and training of the next generation of emergency care clinical trialists. The proposed Upper Midwest hub represents a growth of the prior University of Minnesota hub, with expanding reach and inclusion of sites across the Midwest with a focus on growing rural and teleresearch capacity. Our hub brings academic depth and breadth across wide domains of acute critical care, pre-hospital and emergency research. Hub leadership is experienced in the spectrum of acute care clinical research, pre-hospital research, the use of Exception from Informed Consent for acute critical care clinical trials, positioning us well to conduct all manner of trials supported by the network, while the multidisciplinary specialties needed to collaborate for successful implementation of SIREN are well aligned with collaborative arrangements beyond traditional academic departments.

NIH Research Projects · FY 2026 · 2017-07

PROJECT SUMMARY The goal of this research is to understand how formins shape the architecture and dynamics of the actin cytoskeleton. Formins are a uniquely versatile family of actin regulatory proteins that stimulate both filament nucleation and elongation. Actin filaments assembled by formins are incorporated into a diverse set of higher- order structures that support essential cellular functions, including migration, division, and transport. Mammals express 15 formin isoforms, each of which possesses unique actin assembly properties and plays a specific role in cells. Consistent with this specialization, mutations in individual formin genes are linked to a broad range of diseases and pathologies, including neurological disorders, kidney disease, microcephaly, cardiomyopathy, and several cancers. However, despite their foundational roles as regulators of actin assembly, it is unknown how the polymerization activity of each formin isoform is tailored for the assembly of a specific actin structure. To bridge this gap in understanding, our goal is to establish how the broad range of formin activities influences actin network physiology and dynamics. Our central hypothesis is that formins direct the assembly and specialization of higher-order actin structures by generating binding sites for bundling and severing proteins at isoform-specific rates. We will use a combination of biophysical and cell biological approaches to test this hypothesis by pursuing three specific aims: (1) to elucidate the mechanism that underpins the adaptable polymerization activities of formins, (2) to investigate the effects of formin-mediated elongation on actin filament bundling, and (3) to assess the interdependent contributions of filament nucleation, elongation, and turnover to actin structure dynamics. Our work will establish at a molecular level how formins dynamically regulate the construction, specialization, and function of cytoskeletal structures that are essential for cellular viability and human development. In light of the diversity of formin isoforms, our results will generate fundamental insights into the molecular and temporal regulation of a large number of cellular processes. This will inform and guide our understanding of the molecular pathologies underlying a diverse set of human diseases linked to mutations in formin genes.

NIH Research Projects · FY 2025 · 2017-07

Project Summary The University of Minnesota Training the Next Generation of Surgeon-Scientists in Pancreatology Program will provide independent translational science training experience in pancreatology through a team mentoring-based program for the development of translational science projects as well as University of Minnesota Graduate School curriculum and thesis. The program uniquely focuses on developing professional surgeon-scientists with skills to work collaboratively with PhD and other investigators. A team comprised of an individual surgeon- scientist and a PhD or other appropriate investigator will mentor each resident; both will provide mentorship with the goal of developing surgeon-scientists trained in collaborative, transdisciplinary, translational research. As part of this effort, we will expect all trainees to complete a Masters in Surgical Science. We believe that the combination of these skills and experience within a team science setting will permit the acquisition of skills to effectively function as a surgeon-scientist in the future. Our program areas of strength include nationally and internationally recognized basic and translational research in the areas of acute and chronic pancreatitis, pancreatic cancer, and pancreatic islet transplantation with active involvement in clinical practice. The Executive Committee will provide administrative guidance to the Co-Directors in the selection of trainees and Trainee Mentoring Team; development of the individual trainee program, including an Individual Development Plan; evaluation of trainee and program progress, and strategic planning for future direction. The External Review Committee provides independent, extramural review of the program content and process. In summary, a 2-year dedicated research period, as part of a research training program during the surgery residency is highly likely to increase the number of surgeon-scientists poised to make lasting contributions through collaborative team-based research.

NIH Research Projects · FY 2025 · 2017-07

Project Summary Engineered proteins drive biotechnology and biology as therapeutics, diagnostics, and reagents. While engineering the primary function – e.g. binding – has become relatively robust, identifying proteins that meet the rigors of clinical and practical use remains highly problematic. Many proteins suffer from poor developability – instability, insolubility, low expression, and non-specific binding – that ultimately limits utility. Protein sequence space is immense, and sequence-function relationships are complex. Thus, more efficient methods are needed to map the sequence-developability landscape and reduce the practical burden of identifying developable sequences. Robust, quantitative knowledge of the landscape would [1] empower design of libraries constrained to developable space, [2] enable design of mutants to rescue lead molecules with compelling primary function but developability liabilities, and [3] enhance fundamental insight of factors that dictate protein robustness. Efficient techniques could also [4] enable integrated, upstream library-scale selection for developability. Sequence models are moderately predictive of select metrics but do not robustly quantify the overall landscape. Current experimental approaches are inefficient. Thus, creation and implementation of a platform for library-scale evaluation of protein developability would be transformative to accelerate and streamline the protein discovery and engineering pipeline. We will pursue this objective via three specific aims. Aim 1: Engineer a platform for library-scale evaluation of protein developability. We will develop a set of cellular assays that couple [i] genotype-phenotype linkage, [ii] phenotypic stratification via flow cytometric sorting or growth competition, and [iii] deep sequencing to efficiently quantify metrics of developability for millions of protein variants thereby elevating developability characterization by orders of magnitude relative to current methods. Aim 2: Elucidate sequence/developability landscapes for binder scaffolds. We will quantitatively elucidate sequence-developability landscapes for three ligand scaffolds to [i] empower mutant design to rescue lead molecules with compelling primary function but developability liabilities and [ii] to advance fundamental understanding of the physicochemical principles that dictate protein robustness. Aim 3: Design constrained libraries that yield significantly more developable binders. We will use this insight to design and test constrained combinatorial libraries to yield significantly more developable binders than an unconstrained library. We will test three hypotheses: [i] nested sampling enables the efficient traversal of the sequence/developability landscape to identify an effective constrained library design; [ii] developable space is more evolvable than naïve space (provided library scale diversity is maintained); and [iii] the intersection of developability and evolvability can be effectively identified via these methods.

NIH Research Projects · FY 2026 · 2017-04

PROJECT SUMMARY Most U.S. adults (77%) take dietary supplements (DS) and 87% of them express overall confidence in the safety, quality and efficacy of DS. However, DS are not always safe. DS information is scattered in biomedical literature, social media, and FDA spontaneous reporting system. Thus, to optimize the proper and safe use of DS, there remains a critical need to develop informatics framework with innovative tools and resources to enable us better understand efficacy and safety of DS through multimodal data sources. Built upon our prior project, the objective of this renewal application is to create an enriched DS knowledge base (eDISK) and to develop a translational informatics framework (iDISK-Mine) with innovative informatics approaches to facility DS research using real-world, multi-site EHR data and. We will expand our prior work in two major aspects: (1) Expand our scope to both efficacy and safety (focus of the prior project) of DS using multimodal data sources to enrich our current DS knowledge base (i.e., eDISK); and (2) Develop and evaluate a translational informatics framework to facilitate DS research using multi-site real world EHR data. We propose the following Specific Aims: (1) Create eDISK by integrating DS efficacy and safety from multimodal data sources; (2) Develop a translational informatics framework (iDISK-Mine) to facilitate EHR-based observational DS research; and (3) Evaluate the generalizability and utility of iDISK-Mine on the multi-site EHR data of depression patients. This is the first project to develop a translational informatics framework to advance our DS knowledge using multimodal data sources and enable us to understand how patients (e.g., depression patients) use DS using the real-world EHR data. The successful accomplishment of this project will deliver a novel framework with valuable tools and resources for DS clinical and translational research.

NIH Research Projects · FY 2025 · 2017-04

Abstract Ischemic cardiomyopathy and heart failure are the leading causes of combined morbidity and mortality in humans. Herein, sarcomere dysfunction has a central role in disease pathogenesis. The sarcomere is the essential functional unit of cardiac muscle, directly responsible for the pumping action of the heart. The cardiac sarcomere is a multimeric contractile apparatus consisting of a thin myofilament-based allosteric regulatory complex together with the myosin-based thick myofilament that generates force. Interlacing myofilaments operate in synchrony to regulate and generate the forces necessary for heart performance. Beat-to-beat control of cardiac sarcomere activation refers to the status of the thin filament regulatory system in controlling the degree to which contraction is turned on and off during a twitch. Disruption in sarcomere function underlies the basis for numerous forms of acquired and inherited heart diseases affecting millions of people in this country. Thus, focus here on mechanistic insights into sarcomere regulation underscores the major health relevance of this proposal. Recently, emerging results have come to the fore positing synergistic inter-myofilament regulatory signaling mechanisms, including a new role of myosin cross-bridge ON/OFF states in controlling muscle contraction. Building on our sarcomere activation innovations featuring single unloaded cardiac myocytes, we have made a breakthrough methodological advance, permitting real-time recordings of sarcomere activation in intact cardiac muscle under physiological load. This system is capable of detecting, by intramolecular FRET, multiple myofilament activating ligands during the physiological time course of a single twitch contraction in intact cardiac muscle under load. Guiding hypothesis: During physiologically relevant twitch contractions under load, thin filament activation is controlled dynamically by multiple synergistic inter-myofilament regulatory inputs, including TnC bound Ca2+, TnI switch domain-TnC interaction, OFF to ON state myosin cross-bridges, and MyBP-C in live cardiac muscle. This proposal aims to investigate inter-myofilament signaling by altering the TnI molecular switch mechanism during the physiological time-course of a single cardiac twitch in live cardiac muscles under load; to investigate the mechanism of inter-myofilament signaling during the cardiac twitch contraction in live intact cardiac muscles by modification in myosin cross-bridges; and to investigate the role MyBP-C in inter-myofilament signaling during physiological twitch contractions in intact cardiac muscles. Enabled by our innovative approach, the new insights into inter-myofilament signaling mechanisms gained here will significantly impact our understanding of cardiac function. In turn, this provides the essential foundation to guide new therapeutic discovery for the diseased heart by leveraging the sarcomere as an excellent target for developing new treatments and therapies for the diseased heart.

NIH Research Projects · FY 2026 · 2017-04

Project summary Current theories suggest that mammalian behavior arises from an interaction of different action- selection systems that process information about the past (memory), present (perception), and future (goals to achieve, outcomes to avoid) differently. Current theories dichotomize systems into planning and procedural systems, but newer theories suggest more complex, hybrid algorithms may exist within mammalian decision-systems. Current views of psychiatry are based on dysfunctions in the information processing of those decision systems, which means that treating and alleviating those disorders will be enhanced by a better understanding of the information processing that underlies decision making. A number of disorders (OCD, eating disorders, drug addiction) and a number of RDOC-related dysfunctions (compulsivity, habits, and issues of cognitive and “self-” control) have all been proposed to depend on conflicts between these decision systems. We will build on our established expertise in neural ensemble recording and computational analysis to examine the information processing of decision systems, particularly in questions of conflicts between these systems. Using DREADD manipulation and neural ensemble recording technologies, we propose to identify the mechanisms and computations that underlie action-selection processes under exogenously and internally-driven strategy changes.

NIH Research Projects · FY 2025 · 2017-04

PROJECT SUMMARY Despite advances in treatment options, breast cancer remains the second leading cause of cancer-related deaths in women. Identifying key signaling pathways that drive breast cancer progression is necessary for developing new approaches to target breast cancer. Fibroblast growth factors (FGFs) and their receptors (FGFR) are activated in human breast cancers across subtypes and contribute to breast cancer progression via both autocrine and paracrine mechanisms. The focus of this proposal is to define identify novel mechanisms through which FGFR activation in breast cancer cells contributes to pro-tumorigenic alterations in the tumor microenvironment, which contribute to breast cancer progression. To this end, we have focused on identifying 1) novel transcriptional targets of FGF/FGFR signaling in breast cancer cells and 2) their impact on the stromal environment. Using a model of FGFR-driven mammary tumor growth and progression, we have generate preliminary data that link FGF/FGFR activation in tumor cells with de novo cholesterol synthesis and accumulation. Furthermore, our findings suggest that cholesterol accumulation in tumor cells promotes the generation of an immunosuppressive macrophage population. Although the FGF/FGFR axis has been shown to regulate metabolic functions in some physiological contexts, the link between FGF/FGFR and cholesterol metabolism has not been investigated in the cancer. The studies described in this proposal will test the hypothesis that activation of FGFR in breast cancer cells drives cholesterol metabolism in tumor cells and that these alterations contribute to an immunosuppressive microenvironment. Studies proposed in Specific Aim 1 will the mechanisms by which FGF/FGFR activation in breast cancer cells drives cholesterol accumulation and storage. Studies in Specific Aim 2 will examine the impact of FGFR-driven cholesterol metabolism on the tumor microenvironment. Finally, studies in Specific Aim 3 will use spatial transcriptomics and multiplex imaging techniques to identify links between FGF/FGFR and cholesterol metabolism in human breast cancers. Understanding the mechanisms that contribute to FGFR-driven alterations in cholesterol metabolism in tumor cells and subsequent impacts on the tumor microenvironment will lead to novel therapeutic approaches that target malignant alterations within both the tumor cell and the stroma, leading to enhanced therapeutic efficacy.

NIH Research Projects · FY 2025 · 2016-09

Project Summary I am the Director of the Analytical Biochemistry Shared Resource of the University of Minnesota's Masonic Cancer Center (MCC), a National Cancer Institute-designated Comprehensive Cancer Center. The shared resource has emerged as a leading academic mass spectrometry laboratory, acting as an extension of the MCC's Carcinogenesis and Chemoprevention Program (CCP), and providing critical analytical services to CCP members who account for a large majority of the analysis performed in the facility. There are currently 16 NCI-funded projects involving nine CCP members who use the mass spectrometry facility. The integration of the Analytical Biochemistry Shared Resource and the CCP is such that the mass spectrometry laboratory staff, through extensive training and consultation with group members and collaboration with principal investigators, functions as the analytical component of many of the research groups. The scientific goals of the CCP are to understand the chemical and molecular mechanisms of carcinogenesis. The relevance of this knowledge to public health is through the ability to use it to develop and evaluate practical methods for cancer prevention. I am an integral part of the CCP through extensive consultation and collaboration with the members regarding their mass spectrometry-based analyses, as well as by working very closely with their research groups. I have daily interactions with the investigators and routinely attend laboratory group meetings, contribute to writing manuscripts, and participate in the CCP Seminar Series and the Translational Biomarkers Working Group. The need for my consultation and collaboration with members of the CCP regarding advanced analytical measurement, as well as day-to-day training and oversight of members of their research groups, has been growing steadily with the increasing analytical capabilities of the laboratory and increasing dependency of the users on mass spectrometric analysis to advance their research. There is an ongoing need to continue to advance the analytical capabilities of the facility as the field of biomedical mass spectrometry continues to grow and the research goals of the program members evolve. An external source of financial support would give me the independence and freedom to focus on collaborative efforts with the NCI-funded members of the CCP to provide them with the guidance and knowledge to harness the advanced analytical capabilities of the mass spectrometry laboratory.

NIH Research Projects · FY 2025 · 2016-09

PROJECT ABSTRACT/SUMMARY This is a proposal for the competing continuation of the University of Minnesota T32 Cancer Health Disparities Training Program, funded by NCI since 2011. Disparities in cancer incidence, prevalence, screening, treatment and survivorship are persistent, with persons from socioeconomically under-resourced and disadvantaged populations and racial/ethnic minorities disproportionately burdened. This program meets the essential need to train a diverse cadre of investigators to address cancer health disparities through innovative, multi-level and multi-domain interventions and community-engaged research. The primary goal of our T32 Program is to prepare predoctoral and postdoctoral trainees with the knowledge, skills, and experiences necessary to conduct cancer related intervention research with populations experiencing health disparities. Our 3 Training Aims are to: 1) to increase the number of predoctoral and postdoctoral trainees committed to intervention and translational research addressing cancer disparities by leveraging the strengths of the current training program, including curricular, mentored research, and career development components; 2) each year, prepare three predoctoral and three postdoctoral trainees to reduce cancer related health disparities; and 3) enhance the diversity of the research workforce in cancer related health disparities by committing to recruiting at least 50% of trainees from backgrounds underrepresented in research. We seek to educate researchers who are well prepared to design, implement, and disseminate innovative community-engaged interventions to reduce cancer related disparities. With this renewal, we also will pursue three initiatives that are responsive to trainee feedback and innovations in health equity research. First, our leadership, mentorship, and curriculum have been aligned with scientific approaches to health disparities research focused on multiple levels and domains of influence along the cancer continuum. Second, the role of our Community Mentors has been transformed to facilitate increased opportunities for trainees to acquire skills and experiences in community-engaged research. Finally, we will expand the focus on intervention development to address trainee recommendations for education in intervention implementation. Our training program is designed to prepare investigators skilled in intervention and translational research to reduce cancer health disparities with a focus on community engaged research. We will achieve our goals through an enriching mentored experience, didactic training, and collaborative research training and projects. Overall, our proposal is highly aligned with the NCI priorities for Advancing Public Health in Cancer.