University Of Minnesota

NIH Research Projects · FY 2026 · 2016-09

ABSTRACT Age-related decline in cognitive health is a pressing public health concern. Variation in late-life adult cognitive health is associated with adverse interpersonal experiences (AIEs, e.g., loneliness), although documenting the full extent of this association requires further study of a greater diversity of AIEs. Social isolation and lower quality relationships, characterized by greater conflict, tend to disrupt psychosocial and biological functioning, resulting in increased risk for Mild Cognitive Impairment (MCI), which, in turn, increases risk for Alzheimer’s Disease and Related Dementias (ADRD). Conversely, adults who receive more warmth, support, and responsiveness in their social experiences typically have better cognitive outcomes. This project takes advantage of an unparalleled opportunity to further this important line of work on the significance of relationship experiences in later adulthood for MCI by conducting a follow-up of the Minnesota-Carolina Twin Study of Aging (MCTSA). MCTSA is a longitudinal study targeting 800 ethnically diverse monozygotic and dizygotic same-sex twin pairs. Most twin participants have already completed assessments of AIEs in the previous project period. In this proposed continuation period, we will complete novel assessments of MCI. Further, a daily diary study aimed at connecting AIEs and fluctuations in cognitive functioning on a day-to-day timescale will allow us to document processes that presage the development of Alzheimer’s Disease and Related Dementias (ADRD). Our overarching objectives are to investigate the degree to which diverse AIEs predict subsequent MCI, to take advantage of the twin aspect of our design to document the environmental basis for AIE-MCI associations, and to understand how these connections are manifest in the day-to-day flow of interpersonal experiences and cognitive functioning. Specific aims of this project include: AIM 1: Document longitudinal associations between a comprehensive array of earlier, specific adverse interpersonal experiences (AIE) and subsequent mild cognitive impairment (MCI); AIM 2: Model environmental pathways linking earlier adverse interpersonal experiences (AIE) and subsequent MCI; AIM 3: Complete a twin daily diary study of within-person dynamics connecting adverse interpersonal experiences and daily fluctuations in cognitive functioning.

NIH Research Projects · FY 2026 · 2016-09

Project Summary/Abstract Translating basic vision research to the clinic progresses at an astounding rate, but we still have a long way to go in developing treatments and potential cures for blinding vision diseases. The advancements in methods, e.g., viral vectors, genetic approaches, single cell transcriptomics, imaging modalities, optogenetics, and stem cell-based approaches, have changed our ability to understand the visual system in health and disease. The major aim of the University of Minnesota’s program in the Training in Translational Vision Sciences is to prepare predoctoral trainees for careers as independent scientists with excellent knowledge of basic science, and importantly, provide connections to clinical problems and real patients with vision-threatening disorders. The 18 training faculty are a well-funded, productive group of vision scientists with a commitment to training the next generation of vision scientists. Ten of these 18 have funding from the National Eye Institute, but all preceptors have well-funded laboratories and strong publication records. The major areas of research focus include real- time imaging, visual processing in human and animal models, vascular biology, and development. These focus areas are typically coupled with a strong translational objective focused on developing new strategies to improve vision in individuals with significant visual impairment. We are requesting funding to support five predoctoral candidates who have entered into the laboratories of one of the vision scientist preceptors. Predoctoral trainees will come from one of four graduate programs: “Neuroscience”, “Molecular, Cellular, Developmental Biology & Genetics”, or “Biochemistry, Molecular Biology, and Biophysics” in the medical school, or the Graduate Program in Psychology in the College of Liberal Arts. With the focus on translational vision science, we have greatly increased the collaborations between basic scientists and the clinical faculty in the Department of Ophthalmology and Visual Neurosciences (DOVN). Several new key elements of the Translational Vision Research Training program include a newly designed course for trainees, Critical and Translational Reasoning in Visual Science, co-taught by the training faculty and the clinical faculty in the DOVN. All trainees must attend and present their research at our annual Spring Symposium, where nationally known and in-house experts in vision science present their most recent research along with our trainees and faculty as well as participate in one of several vision-related journal clubs. The Program Directors and Executive Committee select and evaluate trainees for this program. The program directors are Linda McLoon PhD and Harald Junge PhD, both in the DOVN. We have built in a Program Director transition plan, with Drs. McLoon and Junge sharing the position during the first two years of the program, and with Dr. Junge taking over as Principal Investigator in years three to five. As we move toward increasingly collaborative research between our basic and clinically trained faculty members, the training we provide will maximize our ability to prepare vision scientists with a multi-disciplinary education needed to meet the “audacious goals” of the NEI - focused on preventing, treating, and rehabilitating blinding eye disease.

NIH Research Projects · FY 2026 · 2016-09

Project Summary Research in the Tonks group is focused on designing new, enabling oxidative amination reactions using Earth-abundant metals such as Ti and Fe. The goal of our future work is to design catalytic reactions that generate reactive nitrogen equivalents from simple amine precursors. There remains a need for general reactions directly from feedstock amines, rather than high-energy precursors. Such methods are essential to translate new amination reactions to discovery medicinal chemistry, where stable, commercial reagents are necessary, and where convergent multicomponent reactions have strategic advantages over linear syntheses. We will bring to the fore a new concept in organic catalysis, metal-catalyzed multi-site proton-coupled electron transfer (catalyzed MS-PCET), to carry out net dehydrogenative reactions of amines, leading to the reactive nitrene precursors we need for catalytic amination reactions. The advantage of catalyzed MS-PCET is that we can use metal catalysts to circumvent kinetic limitations of H-atom abstraction that are typically tied to the bond dissociation free energy of an N-H bond. This allows us to use mild oxidants and bases to engender net dehydrocoupling or nitrene transfer reactions. Since the catalyst, oxidant, and base are separate reagents, there is a significant opportunity to finely tune reactions and develop/optimize new reactions. Our proposed studies in catalyzed MS-PCET are built from promising preliminary results, where we have discovered an N-H dehydrocoupling reaction catalyzed by FeCl3 that leads to medicinally relevant 2H-indazoles through efficient, mild routes. We will pursue other new inter- and intramolecular N-H dehydrocoupling reactions—focusing on N-N and N-O bond forming reactions because these types of synthetic disconnections are very uncommon but potentially enabling in the synthesis of a wide range of biologically relevant molecules and heterocycles (piperazic acid, indazoles, isoxazoles, for example). A key concept of this work is that ligand oxidation drives productive catalysis through reactive metal-bound radical intermediates. We will explore the photochemistry of metal imido (M=NR) molecules to further extend this concept and enable new electrophilic and biphilic reactions, expanding the scope of our early transition metal amination platform. We will also use our expertise in early transition metal chemistry to develop enabling, bench- stable reagents for high-throughput screening campaigns, filling a key gap in titanium catalytic methods. Relevance to public health. 59% of all unique FDA-approved small molecule drugs contain at least 1 nitrogen heterocycle, making them the most prevalent functional groups. Indazoles and their derivatives are an important class within this group. Although many reactions to form indazoles exist, their synthesis relies on potentially toxic and explosive hydrazines and have well-established regioselectivity limitations. By designing catalytic methods to indazoles, and more generally to the formation of weak N-N/N-O bonds, chemists will have rapid, convergent access to diverse, novel molecules that will aid in in developing new small molecule drug-like architectures.

NIH Research Projects · FY 2026 · 2016-07

PROJECT SUMMARY Cells sense and transmit mechanical forces through cell-surface receptors to orchestrate essential processes, from migration and differentiation to immune activation and chromatin organization, and dysregulated mechanotransduction underlies pathological mechano-phenotypes in cancer, muscular dystrophy, and other disorders. Understanding molecular mechanisms of mechanosensing will lead to understanding of biological functions under tension as well as new therapeutic strategies. Over two MIRA cycles, our lab has advanced three core research programs in the areas of using molecular tension sensors to measure pN forces exerted by cell-surface proteins in cells, using structural biophysics to dissect molecular mechanisms of force sensing and using HUH-tags to enable protein-DNA bioconjugation. In this second MIRA renewal, we ask new questions and continue innovating tools in these three program areas. We will use our high-throughput mechanical profiling assay based on immobilized DNA tension sensors and flow cytometry readout to develop workflows for mechan-omics to link mechanical phenotypes with transcriptomic and epigenetic states and use CRISPR-KO screens to discover mechanosensors unique to abnormal mechanotypes. We will also develop high-throughput luciferase readouts of mechanophenotype and new force sensors to detect inter-cellular forces. We will probe proteolytic switch domains using force spectroscopy and use bioluminescence imaging approaches to study Notch ectodomain architectures and how native versus engineered receptor–ligand interactions impact tension sensing and synapse assembly. Finally, we will mine extremophile Reps for thermostable variants with unique specificity and engineering potential.

NIH Research Projects · FY 2025 · 2016-07

PROJECT SUMMARY AND ABSTRACT The basal ganglia have a rich somatotopy and functional topography composed of motor subcircuits that are thought to be critically important to the pathophysiology of Parkinson's disease (PD) and successful application of deep brain stimulation therapy (DBS) for managing each cardinal motor sign of PD. There is a strong clinical need to better understand these processes and in turn harness them to deliver therapy that is tailored to a patient's own symptomatology and motor control needs on a moment by moment basis. This project will investigate how spatiotemporal optimization of DBS settings can differentially affect neural pathway activation through the brain's motor control network and how those results translate to improving each of the four cardinal motor signs of PD (bradykinesia, rigidity, tremor, and postural instability). Aim 1 will investigate at the single cell, ensemble, and network levels how spatiotemporal parameters of DBS influence information transmission, and critically how the motor control network is able to produce naturalistic movements despite information lesions induced by high-frequency stimulation. Aim 2 will develop and apply a Bayesian Dual Adaptive Control algorithm to investigate how spatiotemporal DBS settings affect electrically-evoked compound action potentials and how those features map onto modulating individual motor signs. Aim 3 will leverage the ground-truth electrophysiological data from high-density microelectrode array recordings at the site of DBS and within the motor control network to validate key parameters used in computational models of neural pathway activation with DBS therapy. The proposed study integrates innovative high-density microelectrode array recordings, closed-loop optimization algorithms, micron-resolution anatomical pathway imaging, and subject-specific computational models of DBS. Together, this project will enhance our understanding of the pathophysiology of PD and provide critical data towards translating next generation personalized and responsive DBS therapies.

NIH Research Projects · FY 2025 · 2016-07

The R25 Summer Research Program in PharmacoNeuroImmunology (PNI) provides up to six meritorious undergraduate students an intensive laboratory-based research experience to increase their participation in biomedical graduate research programs. There is a well-recognized need to train biomedical scientists in an interdisciplinary and translational field that focuses on the interactions of drugs of abuse with the nervous and immune systems and integrates our understanding of these physiological interactions with their behavioral counterparts. We will accomplish this by offering an intensive 10- week research experience that will provide undergraduate students the necessary academic and research skills to ensure their competitiveness toward entering the next phases of their career - i.e., doctoral programs or professional schools- medical, dental or veterinary school. The PNI summer program is part of the comprehensive Life Science Undergraduate Research Program (LSSURP) at the University of Minnesota. The PNI summer program is composed of an intensive laboratory-based research experience that entails close interaction with and mentorship by a program faculty member that focuses on drug abuse research. This laboratory work is supplemented by a comprehensive educational curriculum, which includes seminars by program faculty on topics in biomedical research, ethics, and laboratory safety, and a focused student research poster symposium. An innovative component of our PNI program proposal is to deliver education modules with small group discussion. In addition, Dr. Jesse L. Mason, an alumnus of LSSURP and the T32 PNI training program, will provide a series of sessions for PNI summer students targeted toward dealing with adversity as a graduate student. Three measures of evaluation will be collected on a yearly basis: 1) Assessment of the 10-week summer research experience by participating undergraduate students and faculty in the program. This assessment will come in two ways from the LSSURP survey and a PNI specific survey; 2) The PNI program itself will be supported by the Advisory Committee to verify that we are meeting the goals of the program and 3) Outcome assessment of the students that were in the program. We will track the careers in biomedical sciences of the past summer PNI trainees.

NIH Research Projects · FY 2026 · 2016-07

Abstract Viruses are a major threat to human health. Our laboratory uses various structural biology techniques to dissect molecular mechanisms of how viruses replicate and invade the host cell or its genome. One area of our major interest is retroviral integration, a critical step in the lifecycle of retroviruses that achieves permanent insertion of the reverse-transcribed viral genome into a host chromosome. We will build on our recent structural studies of the Human T-cell Leukemia virus and Rous sarcoma virus intasomes and further investigate the roles of host factors during integration of these retroviruses. Another area that we are pursuing is the replication of coronavirus RNA genomes and host cell invasion. In particular, we are interested in how a virally encoded exoribonuclease complex facilitates faithful replication of the large RNA genomes of coronaviruses, and how this unique proofreading activity could be modulated by small molecules. We are also investigating inhibition of the receptor binding of the coronavirus spike protein by novel antibodies and antibody-mimics. Overall, the studies proposed in this application will help better understand important RNA-based human pathogens and could aid in the development of antiviral strategies, or alternatively, gene delivery tools useful in research or gene therapy applications.

NIH Research Projects · FY 2025 · 2016-07

PROJECT SUMMARY/ABSTRACT With thousands of patients now on the waiting list for a kidney transplant, it is obvious that there is a critical shortage of available donor organs. Xenotransplantation represents a promising solution. This effort is of paramount importance as a significant number of patients are more likely to die waiting rather than receive a life- saving kidney transplant. The major limitation to the clinical translation of xenotransplantation has been the cross-species differences in cell surface carbohydrates/glycans and proteins that serve as targets for recipient- derived antibodies resulting in antibody mediated rejection. We have observed a bimodal distribution of rejection in the first year after pig to NHP kidney transplant with ~50% of recipients treated with our standard anti-CD154 containing immunosuppression regimen experiencing rejection early (<100 days, MST 53 days, n=17) while the majority of the remaining animals survive between 1 and 4.5 years (MST 422 days, n=17). Early graft loss is secondary to antibody mediated rejection directed mostly at glycan related antigens. Late failure has features of chronic antibody mediated rejection and we have shown that this is directed mostly at SLA class II molecules. For kidney xenotransplantation to have clinical utility we need to consistently prevent early antibody rejection while graft survival of at least 3 years in patients to achieve equipoise. In the current proposal we plan to study and address both early and late kidney xenograft failure. As antibody failure remains the most important problem we will address this through data driven genetic engineering to remove known xenoantigens and test novel, clinically relevant strategies to control immune cells responsible for antibody production. We have identified additional glycan xenoantigens that likely contribute to early antibody mediated rejection and have already produced a new pig- GGTA1-/-/B4GALNT2-/-/B3GNT5-/- that we will test in our pig to primate kidney xenotransplant model. We will test whether the elimination of additional candidate antigens will reduce/eliminate the early rejection we see in the first 100 days after transplant and provide consistency towards long-term survival. Late failure in our pig to NHP kidney transplant model is caused by chronic antibody mediated rejection with antibody directed at SLA class II molecules being a major driver. In aim #2 we will use our already generated SLA class II molecule knock out pigs to evaluate the role of SLA DQ and SLA DR in late xenograft rejection. In aim #3 we will target the cells responsible for antibody production using cutting-edge, clinically relevant therapies including a novel anti-CD19 depleting antibody and CD20 and CD19 specific NHP CAR T cell therapeutics to test whether broader and deeper depletion of B cell subsets will prevent early antibody mediated rejection, reset the B cell repertoire, and prevent late antibody mediated injury to the kidney xenograft. The proposed studies address an important clinical problem-- the adequate supply of donor organs as well as critical scientific questions- including what pathways/molecules are essential for the immune response to xenogeneic tissues.

NIH Research Projects · FY 2025 · 2016-06

Inter-organ signals that regulate body size, physiology and developmental timing Identifying and characterizing physiologic mechanisms that regulate organ communication and function during development and adulthood is vital for understanding how many important human health problems arise and intensify during our lifetimes. This proposal outlines a research strategy to further characterize key signaling systems that regulate critical and conserved physiological processes. These include determining how a signaling network of three Drosophila Activin-like members of the TGF-beta superfamily coordinately control fundamental aspects of brain, muscle, and fatbody cellular function in response to environmental variables such as nutrition during development to maintain physiologic homeostasis and how neuroendocrine mechanisms regulate steroid production and release, also in response to various internal and external cues, to properly time developmental maturation. We will use a wide variety of modern biological investigative methods including genetic analysis, neuronal circuit and activity mapping, biochemistry, optical/EM imaging, metabolomics, temporal and tissue specific transcriptome characterization as well as chromatin immunoprecipitation experiments, to answer these questions. Impact on human health: The successful completion of these aims will provide novel insight into how an two very conserved inter-organ signaling networks parse out specific, as well as combinatorial control, over various fundamental cellular processes to produce a properly proportioned animal, that is optimized for survival in its particular ecological niche. It is expected that this knowledge will provide useful paradigms for understanding functional aspects of the more complex, but highly related vertebrate TGF-beta and neuroendocrine signaling systems, and will afford novel insights into molecular mechanisms that contribute to a number of human disorders including obesity, metabolic syndrome, muscle wasting, pubertal disorders and ageing.

NIH Research Projects · FY 2025 · 2016-03

PROJECT SUMMARY Programming of self-antigen specific CD4+ Tregs in the thymus is essential for suppression of aberrant immune responses and prevention of autoimmunity. Tregs also control inflammation during viral infection and following pathogen clearance. This is particularly relevant for some types of viral infections such as the 1918 influenza pandemic strain, and more recently, specific strains of coronaviruses, where the immune response itself can be pathogenic. In addition to viral infections, high levels of type I IFN are a defining feature of some autoimmune diseases, such as SLE or Sjögren’s syndrome; Tregs reduce the symptoms of such autoimmune diseases as well. However, we know relatively little about the Tregs involved in either of the above processes. For example, what types of Tregs are involved in these responses? Do Tregs that dampen anti-viral immune responses or suppress SLE-like disease develop in the thymus? If so, what is the thymic niche that controls differentiation of this Treg subset? What APCs/stromal cells are required for differentiation of this Treg subset? What specific functional roles does this Treg subset play during viral infections or SLE? Our preliminary data demonstrate that a unique subset of Tregs develops in the thymus characterized by a strong interferon-stimulated gene-signature (ISG-Tregs). We propose that this ISG-Treg subset plays a key role in governing antiviral immune responses and suppressing immune responses to autoimmune diseases characterized by high levels of IFN. We will explore this hypothesis in two specific aims. In aim 1, we will determine how IFN signaling in the thymus affects thymic selection and the development of ISG-Tregs. In aim 2, we will determine the functions of ISG-Tregs in response to viral infections and systemic lupus erythematosis. Successful completion of these aims will allow us to identify the mechanism by which ISG- Tregs arise in the thymus, and determine the role of ISG-Tregs in immune responses to viruses, and in autoimmune diseases associated with high levels of IFN, such as SLE or Sjögren’s syndrome.

NIH Research Projects · FY 2025 · 2016-01

Abstract. Natural killer (NK) cells are innate lymphocytes that can be targeted to multiple tumor antigens with exquisite specificity by anti-tumor antibodies, resulting in antibody-dependent cell-mediated cytotoxicity (ADCC). This is a key mechanism of action by several clinically successful monoclonal antibody (mAbs) therapies; however, most patients exhibit or acquire resistance to this immunotherapy. ADCC by human NK cells is exclusively mediated by CD16A (FcγRIIIA), a low affinity IgG Fc receptor. Patient data indicates that increasing the binding affinity between CD16A and antibodies augments the clinical response to therapeutic mAbs. Therefore, we generated a recombinant FcγR consisting of the extracellular region of CD64 (FcγRI), the only high affinity IgG Fc receptor, and the transmembrane and cytoplasmic regions of CD16A, a potent activating receptor. NK cells derived from induced pluripotent stem cells (iPSCs) engineered to express CD64/16A mediated robust ADCC. Moreover, CD64/16A could function as a docking platform for anti-tumor mAbs, arming NK cells with switchable and mixable tumor targeting elements. Our goal is to generate an optimized recombinant CD64 expressed by iPSC-derived NK cells (iNK cells). A compelling scientific premise is provided to support our hypothesis that recombinant CD64 expressed by iNK cells can modulate their activation and enhance their binding to tumor targeting mAbs and cancer cells. Our study will focus on epithelial ovarian cancer, the most lethal gynecologic malignancy, as strategies to enhance ADCC have yet to be carefully investigated. Our hypothesis will be tested by three specific aims: 1) Determination of the in vitro and in vivo ADCC efficacy of iNK cells expressing recombinant CD64; 2) optimization of recombinant CD64 signaling in iNK cells to enhance ADCC and their in vivo durability; and 3) evaluation of the “off-the-shelf” use of iNK cells expressing recombinant CD64 in a preclinical ovarian cancer model, including a high grade serous ovarian cancer patient- derived xenograft. The impact of our study is that it investigates an innovative engineered NK cell platform to express a novel recombinant FcγR to be used in combination with mAb therapies for universal tumor antigen targeting. Our study involves a diverse team of experts with a track record of progressing basic research to the clinic for cancer immunotherapy.

NIH Research Projects · FY 2026 · 2015-09

Abstract Adolescent Brain Cognitive Development (ABCD) is the largest long-term study of brain development and child health in the United States. The ABCD Research Consortium consists of 21 research sites across the country, a Coordinating Center, and a Data Analysis and Informatics Resource Center. In its first five years, under RFA-DA-15-015, ABCD enrolled a diverse sample of 11,878 9-10 year olds from across the consortium, and will track their biological and behavioral development through adolescence into young adulthood. All participants received a comprehensive baseline assessment, including state-of-the-art brain imaging, neuropsychological testing, bioassays, careful assessment of substance use, mental health, physical health, and culture and environment. A similar detailed assessment recurs every 2 years. Interim in-person annual interviews and mid-year telephone or mobile app assessments provide refined temporal resolution of developmental changes and life events that occur over time with minimal burden to participating youth and parents. Intensive efforts are made to keep the vast majority of participants involved with the study through adolescence and beyond, and retention rates thus far are very high. Neuroimaging has expanded our understanding of brain development from childhood into adulthood. Using this and other cutting-edge technologies, ABCD can determine how different kinds of youth experiences (such as sports, school involvement, extracurricular activities, videogames, social media, unhealthy sleep patterns, and vaping) interact with each other and with a child's changing biology to affect brain development and social, behavioral, academic, health, and other outcomes. Data, securely and privately shared with the scientific community, will enable investigators to: (1) describe individual developmental pathways in terms of neural, cognitive, emotional, and academic functioning, and influencing factors; (2) develop national standards of healthy brain development; (3) investigate the roles and interaction of genes and the environment on development; (4) examine how physical activity, sleep, screen time, sports injuries (including traumatic brain injuries), and other experiences influence brain development; (5) determine and replicate factors that influence mental health from childhood to young adulthood; (6) characterize relationships between mental health and substance use; and (7) specify how use of substances such as cannabis, alcohol, tobacco, and caffeine affects developmental outcomes, and how neural, cognitive, emotional, and environmental factors influence the risk for adolescent substance use.

NIH Research Projects · FY 2025 · 2015-09

PROJECT SUMMARY/ABSTRACT The Minnesota HHEAR Targeted Analysis Laboratory’s overall goal is to provide human health researchers with access to state-of-the-art infrastructure for targeted analysis of biological samples. We see this goal as firmly embedded within the concept of the exposome and its relationship to human health. To that end, we are requesting a one-year extension of our project as well as additional funds to complete our assigned HHEAR projects as well as analyze 1800 urinary samples from the Chronic Kidney Disease of Uncertain Etiology in Agricultural Communities (CURE) study for exposure to a number of pesticides.

NIH Research Projects · FY 2025 · 2015-09

The goal of the University of Minnesota Cancer Research, Education and Training Experience (CREATE) program is to train, motivate and prepare the next generation of cancer biology researchers in order to accelerate the reduction of cancer incidence, morbidity and mortality. The CREATE program is an integrated residential 10 week summer research experience that motivates and/or affirms student interest in pursuing a research career, and provides students with foundational skills required for entry into and success in a graduate or MD/PhD program. The foundation of our program is the completion of an independent cancer biology research project under the mentorship of a University of Minnesota Masonic Cancer Center faculty member. We enrich this research experience with regular group meetings to discuss research and develop presentation skills, introduction to core concepts in cancer biology, emerging cancer biology research topics and cancer research career pathways, and the development of professional skills critical for admission to and success in a graduate program. Based on our ability to recruit a talented cohort of students, and outstanding student outcomes in the initial funding period, we are requesting continued support for the CREATE program. We will recruit 18 students each year, with 6 students participating in a novel “MSTP track” where students will be mentored by a physician scientist and participate in additional MD/PhD-specific enrichment activities. The rationale for this new track is based on success with a pilot in the previous funding period, and the need to increase the physician scientist pipeline. For all CREATE students, we have used our evaluation results to revise our cancer biology-specific activities, provide mentor training to all program participants, and focus our professional development activities on supporting and enhancing mentoring relationships, writing personal statements and interviewing. The CREATE program benefits from strong partnerships with the University of Minnesota Masonic Cancer Center and the Medical Scientist Training Program (MD/PhD). We work extensively with the Life Sciences Summer Undergraduate Research Programs, a long-standing umbrella organization that coordinates the activities of several distinct summer research programs at Minnesota. This relationship enhances our recruitment efforts, maximizes administrative efficiencies, and provides additional enrichment activities that complement and enhance CREATE. Continuous quality improvement driven by our evaluation plan will allow us to continue to enhance the student experience and positively impact short-term and long-term measures of success.

NIH Research Projects · FY 2025 · 2015-08

PROJECT SUMMARY / ABSTRACT Fetal alcohol spectrum disorders (FASDs) comprise a range of effects resulting from prenatal alcohol exposure (PAE) including neurological abnormalities, cognitive and behavioral impairments, growth retardation, and craniofacial anomalies. Few treatments have been investigated despite FASD’s tremendous public health burden. Cognitive deficits are a core feature of FASD, and cognition is a natural target for intervention. One potential intervention for cognition in FASD is the essential nutrient choline - known to have effects on brain development and cognition. In the hippocampus, choline contributes to increased dendritic arborization, larger cells, and functional changes. Choline affects the cholinergic system and alters brain structure and function in regions essential for memory functioning, including methylation in the hippocampus and prefrontal cortex. Only a handful of human choline studies for FASD have been undertaken and our group has conducted most of them. Our early double-blind, randomized, controlled trial established safety and tolerability. Our subsequent trial revealed beneficial effects for sequential delayed memory in participants with FASD (greater in younger [ages 2-3] rather than older [ages 3-5] children). Our third (ongoing) study included a long- term follow-up that demonstrated long-term benefits for choline vs. placebo in non-verbal processing, working memory, long-term verbal memory, and ADHD behavior. The proposed studies will include a new clinical trial with a new cohort of 2-5 year old children with FASD. Rather than a placebo- controlled trial, it will be a two-arm block-randomized study with cumulative choline exposure durations of 3 or 6 months. Results will directly inform future clinical implementation of choline as a neurodevelopmental intervention. The proposed studies will also capitalize on three existing cohorts for additional longitudinal studies that will determine durability of effects from early treatment. A 4- year and 8-year follow-up study will each examine cognitive effects as well as structural and functional brain effects using advanced MRI methods. Cognitive measures will include the Stanford- Binet Intelligence Scale, the Elicited Imitation memory test, the NIH Toolbox Flanker test and Picture Sequence Memory Test, and the Minnesota Executive Function Scale. We will examine choline effects on behavior using parent-report (CBCL) and a structured diagnostic interview (KSADS). Select hippocampal sub-fields will be examined for volumetric improvements following choline or placebo. Functional connectivity will be examined and is expected to reflect changes from early choline supplementation. We will also use cortical myelin mapping to evaluate choline’s effect on long-term myelin development. In addition, we will apply diffusion-weighted imaging to determine choline’s effect on white matter microstructure – which is known to be disrupted in FASD.

NIH Research Projects · FY 2024 · 2015-07

Project Summary Lower urinary tract symptoms (LUTS) encompass a variety of bothersome storage and emptying symptoms, including urgency and stress urinary incontinence, frequent and/or urgent urination, nocturnal enuresis (i.e., bed-wetting), difficulty urinating, dribbling after urination, and bladder or urethral pain before, during, or after urination. More than 200 million people worldwide and over 15% of women aged 40 years or older suffer from urinary incontinence. In fact, women are 2-3 times more likely to experience urinary incontinence and 4 times more likely to experience a urinary tract infection in comparison to men. For decades LUTS research has focused on underlying pathology, disease mechanisms, and treatment efficacy. The NIDDK took a bold step by introducing the concept of prevention as an important priority for women's urologic research. Through its transdisciplinary team science approach, the Prevention of Lower Urinary tract Symptoms Research Consortium (PLUS-RC) is creating new conceptual models and paradigms, developing innovative measures, establishing evidence, and leading and seeding research on bladder health and LUTS prevention. As the Scientific and Data Coordinating Center (SDCC) for this unique consortium since its inception, we are excited to further develop the evidence base to empower women and their communities to advocate for an environment that supports healthy bladders. During this next grant period, we plan to (1) provide leadership, management, and biostatistical support in the design and implementation of a nationally and regionally representative prospective longitudinal cohort study to assess the distribution of bladder health and evaluate potential risk and protective factors that influence bladder health status; (2) provide leadership, management, and biostatistical support in the analysis of existing studies (quantitative and qualitative) that will lead to the collaborative development of conceptual models that guide analyses within the observational study above, and new studies that support future primary or secondary prevention initiatives; (3) provide leadership in the principles and process devoted to item development, instrument development, and psychometric evaluation and scoring; (4) facilitate a network of community stakeholders to assist and collaborate in the process of data collection development, execution, and dissemination; (5) foster transdisciplinary team science across all consortium activities and centers; and (6) provide administrative leadership and logistical support and coordination of meetings of the Steering Committee, all subcommittees, the Executive Committee, the External Expert Panel, and the pilot and feasibility studies program, for the efficient and ethical execution of consortium objectives.

NIH Research Projects · FY 2025 · 2015-07

PROJECT SUMMARY The profound detrimental impact to society from individuals abusing drugs is well established. To meet the challenge of developing new and effective treatments to help those that have succumbed to the temptation of drug use and abuse, we need to inspire the next generation of students to pursue research careers in the field. The need is particularly acute among populations of students who are currently underrepresented in the field of drug abuse. Published analyses indicate that exposing undergraduate students, especially early in their career, to laboratory research is an extremely effective way for developing their interest in research as a profession. Since 1989, the University of Minnesota of has recognized and met this challenge by offering summer residential research programs in the biomedical sciences. This proposal is the competitive renewal of a training program that specifically provides research experience in the field of drug addiction. Our goal is to train undergraduate students who have completed their freshman or sophomore years in college. We will recruit students nationally, accepting students from groups that are underrepresented within the biomedical research profession. We will provide them with a 10-week intensive research experience that will include professional mentoring (academic survival skills and preparation for graduate school) as well as workshops on research ethics. Our goal is to inspire a new generation of drug abuse researchers. In turn, we expect these individuals to become part of the research infrastructure dedicated to solving medical problems of nervous system dysfunction

NIH Research Projects · FY 2025 · 2015-07

Project Summary/Abstract The University of Minnesota Medical School is committed to the recruitment, enrollment, research instruction, and mentoring of a diverse group of high-quality medical students with aspirations of a career in academic medicine. Based on surveys of medical students, as well as on an analysis by a faculty-driven Medical Student Research Task Force, we identified research training in the summer between years 1 and 2 as the critical opportunity to capture the interests and talents of academically minded medical students. Thus, this T35 competitive renewal application for a “Medical Student Summer Research Program in Infection and Immunity” represents our plan to continue to provide high quality medical student research instruction and mentorship. Our long-term objective is to significantly increase the number and diversity of medical students that obtain paid employment in medical research, that publish during medical school, and that eventually enter into physician-scientist training pathway residency programs. To this end, we describe herein our activities during the first 5 years of the program (including years 1-4 of this funded T35 grant) and offer the results of our current efforts based on the performance and perceptions of 30 medical student summer researchers. Daniel Mueller, MD, initially developed this Medical Student Summer Research Program in Infection and Immunity, and will continue to lead this T35 training grant. He has a breadth of experience in academic medicine including a) laboratory-based research that includes both fundamental and human-oriented investigation into autoimmune disease pathogenesis, b) medical leadership as a division director, c) an extensive undergraduate and pre-doctoral training record, and d) direct patient care in Rheumatology. Mueller will be assisted by numerous Medical School leaders with records of success in research and mentorship, and will receive additional financial support from the Chair of Medicine and the Dean of the Medical School. A total of 31 outstanding Center for Immunology faculty members will serve as the preceptors for 7 summer medical students in each year of the grant period. Diverse student candidates will be approached by the T35 director on an individual basis, and encouraged to apply to the program. Students entering the program will receive direct instruction in scientific investigation for a period of 2 months. Students will also receive weekly instruction that relates the function of the immune system to human health. Training experiences will include the writing of a formal “Specific Aims Page” proposal, as well as a “Progress Report” final summary of their research. In addition, fellows will prepare and present to Center for Immunology faculty both a scientific poster and oral presentation of their work. Finally, trainees will be given the opportunity for longitudinal mentorship regarding the physician-scientist career path throughout their medical school experience. Success in this program will be monitored both through longitudinal surveys of students and training faculty as well as through oversight by both Internal and External Advisors.

NIH Research Projects · FY 2025 · 2015-06

PROJECT SUMMARY APOE4 is the strongest genetic risk factor for sporadic AD with Ab-dependent and Ab-independent effects on disease pathogenesis. However, the molecular mechanisms underlying the pathogenic nature of APOE4 are not fully elucidated. In previous funding period, we have demonstrated that APOE4-induced phosphoinositol biphosphate (PIP2) dyshomeostasis through the increased expression of a PIP2-degrading enzyme, synaptojanin 1 (synj1). In parallel, a non-biased multiscale network analysis of human dataset from the AMP-AD consortium identifies synj1 as a key driver in both male and female AD subnetworks. Synj1 reduction has been found to provide several beneficial effects such as rescuing APOE4-associated cognitive impairments and lysosomal defects. Notably, we discovered that APOE4+ microglia have higher synj1 expression at baseline, and that APOE4+ microglia manifests with impaired phagocytic activities and lysosomal defects when compared to APOE3+ microglia which can be rescued by synj1 haploinsufficiency. A pilot study indicates that lowering synj1 expression in E4FAD mice protects against cuprizone-induced chronic neuro-inflammation and cognitive/motor deficits in vivo. We postulate that the APOE-synj1-PIP2 signal pathway plays a functional role in regulating microglial function, and that dysregulated microglial synj1-PIP2 homeostasis induced by APOE4 may lead to dysregulated immune function and excessive synaptic elimination, promoting APOE4-associated neuroinflammation and synaptic dysfunction. Therefore, we will characterize the regulation of microglial function by the APOE-synj1-PIP2 signaling pathway during aging and AD pathogenesis in this renewal application. We propose to 1) investigate microglial synj1 function in modulating APOE4-regulated neuroinflammation in AD in vivo (Aim 1) using cuprizone-induced inflammation in male and female EFAD mouse models (human ApoE4 knock-in at 5xFAD background); 2) to characterize the molecular mechanisms by which the APOE-synj1-PIP2 signaling pathway regulates microglial function (Aim 2) using mouse microglia from EFAD mice (synj1+/+ and synj1+/-), mouse microglia from TREM2-/- or APOE-/- mice, and human iPSC-derived microglia from APOE3+/+ and APOE4+/+ normal and AD subjects; 3) to investigate temporal and spatial relationship between dysregulated microglial function and excessive neurite and synaptic elimination using ex vivo 3-D co-culture of iPSC-derived brain cells; 4) to perform high resolution multiscale network modeling using scRNA-seq dataset from mouse brains and RNA-seq dataset from human iPSC-derived microglia and co-culture to identify microglia-specific molecular signatures driven by APOE-synj1-PIP2 signaling; and 5) to validate identified molecular signature in post-mortem human brain samples (Aim 3). The goals of this application aim to elucidate novel pathways and molecular signatures underlying APOE4-induced microglial dysfunction. These studies will facilitate identification and development of a more personalized targeted therapy to APOE-regulated neuroinflammation in AD.

NIH Research Projects · FY 2024 · 2014-11

Using high resolution cryo electron microscopy (Cryo EM) we will investigate the mechanisms used by coxsackievirus B3 (CVB3) to enter host cells by engaging the coxsackievirus and adenovirus receptor (CAR). CVB3 is a human pathogen that causes myocarditis, pancreatitis, and has recently been linked to the onset of juvenile diabetes mellitus. Our proposed studies of CVB3 entry are a continuation of a productive previous granting period. Part of the previous work done was in collaboration with Dr. Jeffery Bergelson, Division of Infectious Diseases at Children's Hospital at Philadelphia, who has since retired. The proposed projects to continue studies of enterovirus entry include a collaboration with Konstantin Chumakov, FDA. Chumakov will work with us to explore one of the most promising results from the previous funding period, which was the discovery that a region in the CAR footprint onto the CVB3 capsid is conserved among three other viruses and their receptors, thus crossing four species using three different host proteins for entry. Further examination showed 1) the region is highly conserved among all human enteroviruses, including all three strains of poliovirus. 2) there are in total four different highly conserved regions. Previous work and our preliminary results link these conserved sites to functions linked to entry. Our structure/function studies will include testing each region for function and as targets for antivirals that are desperately needed. We will use both traditional icosahedrally averaged approaches along with asymmetric reconstruction approaches, some of which have been recently developed by us.

NIH Research Projects · FY 2025 · 2014-07

Project Summary/Abstract This renewal application for the T32 institutional training grant (now in its 10th year), Comorbidity: Substance Use Disorders and Other Psychiatric Conditions (NIDA DA037183), will serve as an essential hub of education, training and career enhancement for a diverse group of clinical, translational, and basic scientists studying comorbidity in substance abuse research and its intersection with other differences (as age, race, and social vulnerability) within the University of Minnesota (UMN) community. The training program provides research training for 4 postdocs annually and has provided training for 13 postdoctoral trainees, with 4 currently in training (17 total trainees). The program's long-term goal is to cultivate a cohort of scientists with research expertise in topics relevant to addiction comorbidity, including mechanisms, antecedents and correlates, diagnostics, and psychosocial and pharmacological interventions. Component objectives are to provide each trainee with a working knowledge of comorbidity research, including (a) translational science from early phase clinical trials to community-based participatory research perspectives; (b) effective research strategies for comorbid conditions across populations and ethnic and cultural groups (e.g., American Indian, Hmong, Somali). A goal of the renewal addresses the growing need for training in addiction comorbidity research across the lifespan. Accomplishing programmatic features will capitalize on (a) the spectrum of faculty expertise providing mentoring across multiple areas and (b) Integration across training programs and departments. Key elements of the program include (1) the involvement of scientists and clinicians with diverse expertise and a core internal advisory group; (2) Recruitment, including underrepresented group outreach, of rigorously screened Ph.D. candidates with SUD and comorbidity as a primary career focus; (3) Training with an interdisciplinary mentoring team with complementary expertise; (4) Formal training plans with clear milestones including trainee development of an NIH application initiated in Year 1; (5) Active research, seminars, didactic coursework, workshops, and development of management, ethics, and regulatory expertise; (6) Dynamic program administration entailing monitoring with enhancements and problem resolution along with continued contact with trainees after completion and; (7) Annually convened advisory committee. Resources: Mentor funding sources include NIH Institutes, NSF, and Minnesota Medical Foundation, Key Personnel and Primary Mentors are directors of clinics, centers, or departments with significant resources.

NIH Research Projects · FY 2025 · 2014-04

Abstract: This renewal continues our optimization of regulatory T cell (Treg) therapy for creating transplant tolerance to prevent graft-vs-host disease (GVHD). Our first-in-human phase I CD4 thymic Treg (tTreg) and CD4 inducible Treg (iTreg) trials that showed reduction but not elimination of GVHD. Importantly, relapse rates were not improved and decreasing GVHD but not relapse is an insufficient outcome. In this renewal, we will build on our exciting finding that Tregs can acquire super-suppressor function and by incorporating an anti-CD19 scFv CAR (CAR19) increase their potency of killing. In other studies, we developed highly suppressive CAR19 CD8 iTregs that have an intrinsic killing mechanism. Comparing the relative efficacy of these cell types, alone or in combination, will allow us to initiate clinical trials using one product simultaneously suppress GVHD and paradoxically kill lymphoma, solving the 2 major alloHCT limitations. A commonality in the diverse approaches and Treg subsets that acquire super-suppressor function is the presence of a striking metabolic effects. We discovered that targeting selected cell surface or intracellular proteins led to mitochondria fusion, high oxidative phosphorylation (OXPHOS) and super-suppressor Tregs. Polar metabolite analytics pointed to key KEGG pathways [serine-glycine-one carbon network], linked by methionine and folate cycles, trans- sulfuration for glutathione, and alanine-aspartate-L-glutamate. A net result is one-carbon (1C) methyl, formyl, or methyl group transfer for biosynthesis, control of gene expression and damaging redox reactions. CAR19 expressing CD4 tTregs and human CD8 iTregs that are highly suppressive and cytolytic to hCD19+ targets will be studied in GVHD/GVL models. Notably, whereas activated CAR19 CTLs can cause severe cytokine release syndrome (CRS), our preliminary data indicate that CAR19 CD4 tTregs produce low inflammatory cytokines and do not cause CRS. Aim 1A tests the hypothesis that metabolic reprogramming of Tregs depends on purine synthesis, mitochondria fusion and OXPHOS, and glutathione for redox balance. Aim 1B exploits Tregs with 4-1BB vs CD28 and CD2) signaling domains that affect metabolic reprogramming and Treg persistence for optimal GVHD/GVL. Aim 2 is focused on the killing mechanisms that these Treg subsets use to lyse targets. Toward that end, we recently discovered CTL supramolecular attack particles (SMAPs) as a new mechanism for more sustained killing. SMAPs are cytotoxic core-shell particles that contain granzyme B and perforin and are secreted after triggering the immunological synapse (IS), the contact signaling point for T cells and APC. Identifying integrated IS signals for SMAP release will provide insights into Treg killing mechanisms, aiding Aim 1. Supported lipid bilayers simulating the IS will assess CD2 as an IS signal integrator/amplifier of killing by analyzing dual CAR Tregs. Our central hypotheses are that Tregs reprogrammed in aim 1 for 1C-, purine- and glutathione- metabolism have enhanced suppression and SMAP release, triggered by CD2 integrated and amplified IS signals and optimal GVHD/GVL.

NIH Research Projects · FY 2025 · 2014-04

ABSTRACT The long term goal of our research is to understand the computational role of mental imagery in human cognition, and to operationalize this understanding to provide new tools for improving mental health. For most people, mental imagery is experienced as visual content that is independent of vision but indispensable for thought. This experience suggests that mental imagery serves an important cognitive function; however, studies of the brain systems that generate mental imagery have yet to reveal how or if mental images contribute to cognition. In this proposal, we test the hypothesis that mental imagery supports cognition by permitting the comparison of seen to unseen images. The comparison of seen and unseen images is a routine operation that occurs, for example, when one judges how a seen image differs from a remembered image, or from a target image that one wants to detect. To test this hypothesis we will measure brain activity in people as they imagine and as they complete a variety of tasks that require them to compare pictures displayed on a screen to pictures that they have been asked to remember. Using techniques borrowed from artificial intelligence (AI) we will extract from these data information about individual mental images, and then determine if this information can predict brain activity and behavior during the tasks. If successful, this proposal will establish a functional and computational role for imagery. Since very little is currently known about the function of mental imagery, our work is an essential step toward understanding how mental imagery interacts with and supports cognition, and how disregulated mental imagery can disrupt mental health.

NIH Research Projects · FY 2025 · 2013-09

ABSTRACT Central nervous system (CNS) infections are common across all ages in Africa in people with or without HIV-infection. In people with HIV, cryptococcal meningitis has historically been the second most common AIDS-defining illness in Africa and the most common cause of adult meningitis in Africa overall. The next most common cause of meningitis is likely TB meningitis, although CSF diagnostics are challenging. With the widespread availability of antiretroviral therapy (ART), long term survival in persons living with AIDS and CNS infections should be possible, but delayed or inaccurate diagnoses and limited therapeutic options contribute to poor outcomes. In the era of widespread ART access, more people are presenting with CNS infections unmasked after starting ART or with virologic failure, yet their epidemiology and outcomes are unclear. We propose to continue a prospective cohort study to enroll >1000 new persons presenting with CNS infections and HIV in Uganda. We will use point-of-care and molecular diagnostics to rapidly determine the etiologies of CNS infections, with a specific focus on optimizing and validating new diagnostic tests, such as a semi-quantitative cryptococcal antigen (CrAg-SQ) lateral flow assay and Xpert-HR Host Response gene signature for active tuberculosis. Second, we will investigate a new oral antifungal, oteseconazole, conducting a phase II randomized proof-of-concept clinical trial to investigate microbiologic activity in cryptococcosis. Oteseconazole is a tetrazole that is >100-fold more active in vitro than fluconazole. Third, we will determine the impact of oteseconazole on neurocognitive performance vs standard-of-care. Our Specific Aims include: 1. Determine the etiology of CNS infections in Africa among HIV-positive adults through use of a stepwise diagnostic algorithm, accounting for HIV therapy status. 2. Determine if adjunctive oteseconazole has greater in vivo activity than adjunctive fluconazole + flucytosine when added to single-dose IV liposomal amphotericin B in HIV-associated cryptococcal meningitis. 3. Determine if oteseconazole is associated with better quantitative neurocognitive performance Z-score (QNPZ-8) at 3-months after cryptococcal meningitis compared to standard fluconazole consolidation. Hypotheses: 1. We hypothesize that in the ART era, cryptococcal and TB meningitis will remain the two most common etiologies of meningitis in an HIV-positive population, with the majority now presenting ART-experienced. 2. We hypothesize oral oteseconazole will have a superior quantitative rate of CSF sterilization when added to IV amphotericin in comparison to adjunctive fluconazole + flucytosine. We hypothesize oteseconazole will have a lower incidence of culture-positive relapse through 1 year of follow up than standard fluconazole. 3. We hypothesize oteseconazole will have better QNPZ-8 at 3-months vs those randomized to fluconazole.

NIH Research Projects · FY 2025 · 2013-04

Recessive dystrophic epidermolysis bullosa (RDEB) is a rare inherited mucocutaneous disease caused by mutations to the COL7A1 gene. Advancements in gene therapy have opened avenues for treatment by introducing a functional copy of the mutated gene in the patient’s cells. Viral vectors, gene editing nucleases, base editors, and prime editors are potential strategies for restoring expression of COL7A1. Lenti-, retro-, and herpes simplex virus (HSV) approaches are all being employed clinically. Ex vivo cell correction with integrating vectors (i.e. lenti- or retroviral) requires specialized facilities for engineering cells and these systems pose insertional mutagenic risks that are a consideration in the RDEB phenotype that has a predisposition to aggressive squamous cell carcinoma development. HSV-1 is a non-integrating vector that reduces genome toxicity related safety concerns but episomal nucleic acid could be lost over time. All of these viral vector systems mediate supranormal gene expression levels. In situ, locus specific correction is desirable as it retains the endogenous gene regulatory network and minimizes insertional mutagenesis. Here we will evaluate highly promising genome engineering and delivery approaches as part of a strategy to restore COL7A1 expression in cells ex vivo and in vivo. In the first aim, we will define and optimize evolved engineered Prime Assisted Site- Specific Integrase Gene Editing (eePASSIGE), a powerful addition to the genome engineering armamentarium. Polymeric nanoparticles (PNP) will be employed for the delivery of eePASSIGE nucleic acids that will install a full length COL7A1 cDNA into the endogenous locus ex vivo and in vivo. Mesenchymal stromal cells (MSC) that express ATP-binding cassette superfamily member (ABCB5) display homing and tissue repair properties and we will evaluate the ability of eePASSIGE corrected autologous ABCB5+ MSC to correct RDEB pathology in mice in vivo. In the second aim, we will develop a novel microfluidics-based epidermal poration device to deliver eePASSIGE PNPs topically to mice for in vivo, permanent COL7A1 gene correction. Both aims will then be synergized to assess the combinatorial potential of topical and cell-based therapy to ameliorate the cutaneous and internal disease manifestations toward filling a therapeutic gap in current treatment options. Our proposed experiments will develop a novel ex vivo and in vivo genome editing strategy that could be broadly applied across all RDEB patient cohorts and that will enhance overall efficiency and correct the full spectrum of the disease.