University Of Minnesota

NIH Research Projects · FY 2025 · 2021-08

Title: Leveraging human evolutionary history to improve our fundamental understanding of complex disease architecture Abstract: The overarching goal of this research is to improve the applicability of genetic risk predictions within and across human populations by leveraging recent advances in our understanding of human evolutionary history. In Aim 1, I will carry out empirically-guided simulations to investigate how fine-scale substructure in large genome-wide association studies (GWAS; e.g. UK Biobank) biases our inference of complex trait architecture and polygenic risk score prediction. I will leverage these findings to develop statistical and computational tools to correct for such biases. In Aim 2, I will investigate whether recent admixture in humans generates incompatibilities between mitochondrial and nuclear DNA in African Americans. To test this, I will analyze genetic and electronic health record data from the ethnically diverse Penn Medicine Biobank to test whether mito-nuclear discordance—degree of ancestry divergence between mitochondrial and nuclear genomes—is associated with the risk of diseases common among African Americans. Additionally, I will test for selection against mito-nuclear incompatibilities in recently admixed populations. In Aim 3, I will investigate how the practice of endogamy and consanguinity among Pakistanis shapes their disease risk architecture. I will further evaluate the ability and limitations of currently used GWAS methods, which are typically modeled after outbred populations, to infer disease architecture given the complex population structure in Pakistanis. I will improve upon these methods, thereby making GWAS more widely applicable to a diverse set of people. Each of my three aims is independent, yet together they will lead to improvements in diagnosis, treatment, and prevention of human diseases—the overarching mission of the NIGMS. I will learn the skills needed to accomplish these aims with the help of my advisory committee, comprising of Drs. Iain Mathieson, Sarah Tishkoff, Doug Wallace, and Marilyn Ritchie, who are world-class leaders in genetics research. With the training plan that I have outlined and the resources at the University of Pennsylvania, I am confident that the K99 award will help me achieve my goal of becoming an independent scientist in the field of statistical genetics.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT The remarkable efficiency of human perception derives from the fact that we do not process each stimulus as a novel event. Instead, past experiences and scene context inform internal, working models of the world that allow us to generate predictions for our physical environment. A leading theory suggests that perceptual predictions are accomplished via flexible normalization: local inhibitory neuronal populations are regulated by long-range connections so that responses are suppressed when they do not provide helpful information about object boundaries. However, the precise neural mechanisms by which the healthy human brain accomplishes this flexible normalization are not known. In order to understand exactly how neural population responses are suppressed or enhanced in response to different scene contexts, we will perform 2-photon imaging in ferret primary visual cortex (V1) to quantify the responses of excitatory and inhibitory neural populations in superficial layers of cortex during several different visual stimulus paradigms. The ferret model is chosen because the imaging techniques necessary to quantify inhibitory neuronal responses are not yet well established in primate models, and while our current knowledge about neural morphology and connections has been derived from mouse models, mouse visual cortex lacks the “columnar organization” (spatial grouping of neurons with similar response properties) that is a hallmark of primate visual cortex and is present in ferrets. Thus, the ferret model is well-positioned to bridge the gap between mouse models and primate models. First, in order to understand neuronal behaviors in the absence of contextual modulation, we will characterize interactions within a single hypercolumn to small, simple stimuli (sinusoidally modulated luminance gratings) at a range of orientations and contrasts. We hypothesize that parvalbumin-containing (PV+) inhibitory interneurons will demonstrate the sharpest orientation tuning, followed by somatostatin-containing (SOM+) and serotonin-positive (5HTR+) populations. Next. using a Cross Orientation Suppression paradigm, we will test the hypothesis that that SOM+ responses track the overall contrast energy in the stimulus, while PV+ populations reflect suppression of individual grating component representations. Additional experiments with naturalistic textures will test whether these behaviors generalize to stimuli with a broad range of contrasts, orientations, and spatial frequencies. Finally, we will use classical Orientation-Dependent Surround Suppression and Collinear Facilitation paradigms to study how the local inhibitory pool responds to scene context. We hypothesize that the responses of local 5HTR+ neurons will reflect the surrounding stimuli rather than the center stimuli. Together, these experiments will constrain an open-source computational model articulated at the level of the single neuron that will constrain hypotheses about how human perceptual behaviors are linked to specific neuronal populations; this model will be valuable for understanding how perceptual aberrations associated with psychosis might be mapped to the function of specific neuronal subpopulations.

NIH Research Projects · FY 2025 · 2021-08

Abstract Acute leukemia is the most common childhood cancer, and remains one of the leading causes of death in children under 15 years of age. Prevention, therefore, remains the ultimate goal. An essential part of future preventive efforts will involve identifying children harboring preleukemic clones at birth. Several leukemia-initiating genetic lesions have been shown to arise prenatally; however, many other leukemia-initiating lesions have not been examined at birth. Several critical questions remain to be answered: 1) What subtypes and what proportion of childhood leukemia develops in utero? 2) What is the specific cell(s) of origin in which preleukemic clones arise? and 3) Are some newborns more prone to developing leukemia-initiating lesions than others? In this project, we will answer these questions, with a focus on the ~70% of patients with known translocation or point mutation driver events. This project has three main aims. Aim 1: To identify the presence and clonal frequency of prenatal leukemia-initiating lesions at birth. We will obtain matched cord blood (CB) and diagnostic leukemia samples from ~250 childhood leukemia patients in the Children’s Oncology Group Project:Every Child study. Backtracking will focus on ~182 patients with translocation/mutation-driven subtypes. We derive patient-specific somatic mutations from tumor profiling, then use droplet digital PCR (ddPCR) in flow-sorted CB cells to confirm the presence and frequency of leukemia-initiating lesions at birth, and at different hematopoietic stages. We will also use ddPCR to backtrack lesions in available newborn dried bloodspots (N ~45). Aim 2: To determine the cell of origin of leukemia- initiating lesions, the transcriptomic changes from preleukemia to overt leukemia, and whether secondary mutations arise prenatally, across childhood leukemia subtypes. In 50 childhood leukemia patients where CB ddPCR is positive we will conduct single-cell TARGET-seq on flow-sorted cell populations to simultaneously detect the presence of each patient-specific leukemia-initiating events, secondary mutations, and gene expression in genotypically- and immunophenotypically-defined populations at a single cell level. Aim 3: To determine whether the presence and frequency of preleukemic clones correlate with known risk factors for leukemia. Demographic, perinatal, and genetic risk factors for childhood leukemia will be obtained through parental survey, birth records, and sequencing data. For overall ALL and AML, and for common subtypes, we will test for association between risk factors and the presence and clonal frequency of preleukemic clones at birth, as measured in Aim 1. Identifying a specific cell of origin of preleukemic genomic alterations, and their frequency in neonatal blood, will shed light on childhood leukemia etiology and have important implications for precision prevention efforts. This study will identify key steps required for leukemogenesis by directly comparing the genetic and transcriptomic architecture of preleukemic CB to the subsequent full-blown leukemia. Results will provide a platform for development of large- scale population testing of preleukemic clones in healthy newborns and will enable the first cohort studies examining progression from pre- to overt leukemia.

NIH Research Projects · FY 2025 · 2021-08

Abstract In Sub-Saharan Africa, tuberculous meningitis (TBM) is the second most common cause of adult meningitis, and a major cause of morbidity and mortality among people living with HIV. While host immuno- deficiency clearly drives TBM pathogenesis, pathologic immune responses can also worsen disease. The key drivers of HIV-associated TBM pathogenesis remain undefined but likely differ from HIV-negative TBM, thus a study of the pathogenesis of TBM in HIV-infected humans is warranted and innovative. Opportunities for host-directed therapy in this vulnerable population remain unexplored. To optimize treatment of HIV/TBM and improve survival, it is critical to fully characterize host responses at the site of infection and identify immune signatures associated with good or poor outcomes. To this challenge, we bring our skills in experimental immunology of tuberculosis, matched with an experienced research team with a proven track record of clinical and translational research regarding AIDS-related meningitis in Uganda. Diagnosing TBM is notoriously difficult. The poor sensitivity (~50%) of standard methodologies detects only a subset of those with TBM, likely with the highest CSF bacillary burden. In these patients hypo-functional or pathologic immune responses, representing opposite extremes of immune function, may contribute to poor host control of infection. The higher sensitivity of Xpert Ultra enables semi-quantitative diagnosis of those with a lower burden of CSF bacteria and identifies a group with better immune control of the infection. Our preliminary data suggest that diagnosis with trace or very low Xpert Ultra is associated with better survival. In this project, we propose a new microbiologic/immunologic framework for understanding TBM, categorizing patients based on the differing Xpert Ultra PCR cycle-threshold, which serve as a surrogate for CSF bacterial burden. We seek to interrogate this framework by defining disease outcomes including survival and neurocognitive testing in these different framework groups, while correlating these findings with immunologic analyses of cellular immune responses in the CSF. Our central hypothesis is that CSF immune signatures correlate with key aspects of TBM disease pathogenesis including sensitivity of diagnostics, disease outcomes, and treatment responses. To test this, we will perform high parameter spectral flow cytometry and multiplex cytokine profiling of samples from the CSF and autopsy specimens of patients with HIV/TBM. By comparing these comprehensive immunologic data in groups of patients with either high or low CSF bacterial burden, in those with good or poor outcomes, and in the context of a clinical trial of standard vs high dose rifampin treatment, we aim to define the key contributions of host immunity to TBM pathogenesis. If our hypothesis is correct, the implications of this research are that immunomodulatory therapy will need to be customized to address the paucity or excess of immune responses.

NIH Research Projects · FY 2025 · 2021-08

Arboviruses such as, Dengue (DENV) and Zika (ZIKV), have been associated with several global epidemics. DENV infections can result in a mild fever to potentially fatal dengue shock syndrome, dengue hemorrhagic fever and death, while ZIKV infections can result in congenital microcephaly and the development of autoimmune diseases. Initial clinical manifestation of DENV and ZIKV disease are similar and co-infections have been reported. Most challenging, DENV and ZIKV are known to co-circulate, increasing the probability of co-infections, and thus the potential for severe disease. Consequently, since there is no effective vaccine or form of treatment for either virus, the development of anti-ZIKV and anti-DENV therapeutics is an urgent unmet medical need. Over the past decade prodrugs of nucleotides, referred to as ProTides, have been found to have potent antiviral activity. The anti-hepatitis C virus (HCV) drug sofosbuvir (FDA approved 2013) is the most notable ProTide success, since its use in combination therapy can clear Hepatis C virus infections. Recently, a similar ProTide, remdesivir, has been approved by the FDA, Japan, the European Union an Australia for the treatment of severe SARs-CoV-2 infections by intravenous injection. Carboxyesterase-anhydride-HINT1 (CAH) ProTides, such as sofosbuvir and remdesivir, utilize a four step activation process based on initial esterase hydrolysis, carboxylic acid nucleophilic attack at the phosphorous and release of phenol, followed by mixed anhydride chemical hydrolysis, and finally phosphoramidate hydrolysis by hHINT1. Unfortunately, although they can be dosed intravenously, due to high first pass metabolism, CAH-Protides are limited in their use as oral drugs to viral hepatitis. Furthermore, due to the high carboxyesterase activity of rodent plasma, it has been difficult to carry out pre-clinical studies with rodents and translationally relevant dosages of CAH-ProTides. To address the inherent issues surrounding current ProTide approaches, our group has designed an alternative ProTide approach, we refer to as anchimerically HINT1 activated proTides (AHA-ProTides). Our laboratory has demonstrated that AHA-proTides can be designed that are stable, orally bioavailable, cell permeable and dependent on HINT phosphoramidate hydrolysis. Recently, we have also demonstrated that AHA proTides can be prepared with enhanced potency against DENV and ZIKV. Consequently, we propose to the design and develop anti-DENV and anti-ZIKV AHA ProTides that can serve as advanced lead compounds for the potential treatment of both DENV and ZIKV disease.

NIH Research Projects · FY 2024 · 2021-08

Vaccination is currently the best method for preventing influenza virus infection and for reducing disease severity. Unfortunately, current vaccine regimens suffer from several major drawbacks and efficacy can vary dramatically year to year. Improved vaccine strategies are desperately needed. Preclinical testing is critical for evaluating potential safety and immunogenicity of next generation influenza virus vaccine candidates. Mice are an ideal first model organism because there are a wealth of reagents and genetic tools allow refined experimental approaches and the capacity to do rigorous mechanistic studies, longitudinal kinetic analyses, sampling of mucosal tissue, and assessment of protection through lethal challenge, all at low cost. While mice are not natural hosts for influenza virus infection they can still recapitulate many aspects of human disease. Additionally, some strains are directly pathogenic while others can be readily mouse adapted. Unfortunately, many therapeutic interventions that were successful in mice have failed to translate to humans. This could be due to several factors including species genetic differences and/or environmental factors. We have previously demonstrated that exposing SPF laboratory mice to diverse pathogens from pet store mice recapitulates many of the human cellular and molecular immune signatures absent in standard mouse models. Preliminary studies demonstrate that heterologous protection and influenza-specific serum antibody isotypes are dramatically altered in ‘dirty’ cohoused (CoH) compared to SPF mice, which more closely resemble what has been observed in humans. We hypothesize that CoH mice will better predict immune responses to influenza infection and vaccination compared to standard mouse models. We will test this hypothesis in three aims. Aim 1 will compare the immune response to a panel of adjuvants used in humans between SPF and CoH mice. Where available, we will compare transcriptional profiles to human data. Aim 2 will determine how a diverse infection history impacts the generation, function, and durability of influenza-specific memory T cells, including extensive analyses of lung immune surveillance.to influenza virus infection and vaccination. Aim 3 will determine the immune response to live attenuated, seasonal split and adjuvanted split vaccinations in SPF and CoH mice. This aim will also evaluate the immune response to less immunogenic targets of universal influenza vaccines. Collectively, this proposal will rigorously evaluate the immune response to influenza virus infection and vaccination in mice with diverse infection histories and immune signatures that more closely align with humans. We propose that the intrinsic advantages of the CoH mouse model will significantly complement ferret or other large animal models to improve the pipeline for preclinical testing and enhance translation potential of next generation influenza virus vaccines.

NIH Research Projects · FY 2025 · 2021-08

Project Abstract/Summary Development of type 2 diabetes (T2D) prior to the age of 20 years has been associated with rapid disease progression and early exogenous insulin dependence.1 Furthermore, adolescents with T2D are more likely to develop diabetes-related comorbidities, such as hypertension, atherosclerosis, and kidney disease earlier compared to adults, highlighting the need for a fundamentally different (and perhaps more aggressive) treatment approach in adolescents.1 Obesity (body mass index [BMI] >95th percentile for age and sex) is a primary risk factor for the development and progression of T2D.2,3 However, current treatment guidelines for T2D in adolescents recommend lifestyle management and metformin (+ insulin) as first-line therapy, which rarely result in BMI reduction or slowing of T2D disease progression.1,4-7 Therefore, novel treatments that meaningfully reduce BMI, delay exogenous insulin dependence, and potentially slow the progression of T2D need to be investigated in adolescents with T2D. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) may represent an ideal first- line therapy for adolescent T2D. GLP-1 RAs increase postprandial insulin secretion and reduce glucagon production, and at higher doses, can result in clinically meaningful BMI reduction by suppressing appetite and enhancing satiety.8 Liraglutide (a daily injectable GLP-1 RA) at its 1.8 mg/day dose was approved for adolescents with T2D in 2019, but liraglutide has not been studied with a primary focus on BMI reduction and insulin sensitivity or ß-cell function in adolescents with T2D.9,10 Therefore, the overall objectives of this study will be to 1) evaluate the effects of liraglutide at its obesity medicine dose (3.0 mg/day) versus standard-of-care on BMI reduction, and 2) evaluate its effect on insulin sensitivity and β-cell function in adolescents with T2D and obesity. The overall hypothesis is that liraglutide 3.0 mg/day will result in a greater mean BMI percent change as well as improvements in insulin sensitivity and β-cell function as compared to placebo plus standard-of-care. The focus on using liraglutide at its obesity medicine dose in adolescents with T2D is novel and important; prior studies evaluating liraglutide have not been generalizable to adolescents with this aggressive disease nor have prior studies had the majority of patients on the maximum liraglutide dose, making dose-dependent weight-loss evaluations difficult. The proposed study will generate preliminary data to inform the design of a larger and sufficiently-powered R01 trial. Importantly, this mentored project will provide essential training in clinical trial design and implementation in a unique and challenging patient population, as well as measures of insulin sensitivity/β-cell function. These skills will facilitate my scholarly independence and serve as the foundation of my future career focusing on the application of obesity medicine principles in the treatment of adolescents with T2D.

NIH Research Projects · FY 2025 · 2021-08

Project Summary The proposed study is a randomized clinical trial that uses a Sequential Multiple Assignment Randomized Trial (SMART) study design. The study will directly compare the efficacy of an innovative intervention that combines explicit and implicit approaches to a traditional implicit treatment approach to teach true grammatical forms to children with developmental language disorder (DLD). The study will also compare interventions that include sequences of Explicit- added and Implicit-only treatments. Participants will include 5- through 8-year-old children with DLD who present with significant grammatical weaknesses. In Phase 1, 155 participants will be randomized 1:1 to an Explicit-added treatment group or an Implicit-only treatment group. Each participant will complete 32 sessions targeting four unique grammatical forms (8 sessions/form). In Phase 2, “Masters” will be re-randomized to receive no treatment 32 sessions of the same treatment, or 32 sessions of the alternative treatment. “Non-Masters” will be re-randomized to receive 32 additional sessions of the same treatment or 32 sessions of the alternative treatment. Performance will be measured on acquisition, maintenance, and generalization probes obtained immediately,1-, 6-, and 12- months post-intervention. The SMART study design will be used to determine if child factors, including expressive and receptive language abilities, nonverbal IQ, and executive function skills can reliably predict the treatment sequence that optimizes language learning. Study results will help to determine the best sequence approach to ameliorate grammatical weaknesses, one of the core deficits of young children with language impairment.

NIH Research Projects · FY 2024 · 2021-08

Project Summary Inward Rectifier K+ channels (KIR) play key roles in the operation of cells in neuromuscular and other tissue. Pathogenic variants are linked to numerous neurological, cardiovascular, and metabolic disorders. Although some variants cause gating defects in KIR by altering ligand regulation or ion permeation, there is growing evidence that many –perhaps most– variants cause defects in folding and trafficking of KIR. Despite the central role for folding and trafficking in the disease etiology, there have been to date no comprehensive large-scale studies that determine sequence and structural determinants of KIR trafficking and functional robustness. Here we provide the first comprehensive assessment of missense and topological mutations’ effects on KIR trafficking and function. Acquisition of these data is the required first step to build quantitative biophysical model of the sequence, structure, and function relationship in KIR. These models will be useful to understand the mechanistic basis for KIR mutation phenotypes, to predict their pathogenicity, and to identify new treatment strategies for KIR- linked disorders. These models will also pave the way for rational engineering of KIR as chemogenetic reagents that can be used to study functional roles of K+ channels in intact tissues.

NIH Research Projects · FY 2025 · 2021-07

ABSTRACT This K01 research and training grant aims to use innovative value of information (VOI) analysis methods to efficiently inform the prioritization and design of potential clinical research studies related to managing virologic failure on dolutegravir (DTG)-based ART for adults (15 years and older) in Kenya, which will also have broader implications for resource-limited settings (RLS). The introduction of generic DTG to RLS raises new questions about optimum clinical management for people living with HIV for whom DTG-based regimens are failing. Conducting furtherclinicalresearch can informand improve HIV treatment guidelines; however, research funding resources are limited. Thus, it is important that methods for setting funding prioritie s maximize the potential downstream health benefits of research. The proposed K01 will leverage an individual-based HIV epidemiologic stochastic model, developed by Dr. Duarte, for a Kenyan adult (15 years and older) population. This project’s research aims (R1-3) are to: R1) Given currently available data, evaluate the potential cost-effectiveness of drug resistance testing to inform choice of second-line ART for patients with unsuppressed viral load on DTG-based ART; R2) Calculate the maximum potential returns on investment to be expected from further research related to managing virologic failure on DTG-based ART and identify which research studies have the greatest potential health benefit; and R3) Calculate the optimal sample size for potential key clinical studies identified in Aim 2. The research aims of this study will provide Dr. Duarte the opportunity to pursue his training aims ( T1-4), including: T1) Develop expertise in Bayesian theory and methods for model parameterization and calibration, conducting VOI analyses, and designing clinical studies; T2) Develop expertise in VOI analysis, including statistical methods for meta-model development; T3) Develop expertise in using open-source decision analysis methods, including data visualization tools; and T4) Understand the methodologic and pragmatic aspects of clinical study design, and develop relationships with stakeholders in Kenya and other RLS, in order to build research collaborations. These research and training goals will provide Dr. Duarte with valuable expertise that is applicable to a wide range of important health policy issues relevant to HIV, as well as other infectious disease challenges, thereby preparing him to transition to independence. He will plan to submit an R01 grant focused on informing the prioritization and design of clinical research to improve pediatric and maternal HIV health outcomes in RLS. This R01 will leverage the expertise Dr. Duarte gains through the proposed K01 and combine it with his ongoing, pilot grant-funded work building simulation models for pediatric and maternal HIV-affected populations. Dr. Duarte will complete his K01 career development training at the University of Washington, which provides an outstanding environment for training in computational methods and applying them to global health issues, as the university has a strong >25-year history of collaboration with Kenyan institutions.

NIH Research Projects · FY 2025 · 2021-07

SUMMARY This application will provide the Principal Investigator, Dr. Pamela Lutsey, with protected time and other support to 1) accelerate a successful record of mentoring junior clinical investigators and graduate students engaged in cardiovascular disease (CVD) patient-oriented research (POR), 2) obtain additional training in pharmacoepidemiology, -omics and mentoring best practices and 3) advance her ongoing CVD pharmacoepidemiology research program. Pharmaceuticals play a crucial role in the prevention and treatment of CVD and other conditions. To improve the effectiveness of pharmacologic interventions on CVD outcomes, we need junior clinical investigators conducting POR who have rigorous training and mentorship in the areas of pharmacoepidemiology, clinical epidemiology of CVD, translational research methods and the use of complex data sources. An expert in these areas, the candidate, Dr. Lutsey, proposes to apply K24 funds toward the mentorship of POR scholars at the University of Minnesota who are committed to a research career that will reduce the burden of CVD in the population through building rigorous evidence regarding how to optimize use of pharmaceuticals in order to prevent CVD in at-risk populations and reduce risk of recurrence among individuals with CVD. Aim 1 involves conducting pharmacoepidemiology POR comparative effectiveness research to prevent CVD in at-risk populations and improve the outcomes among CVD survivors. State-of-the-art pharmacoepidemiologic techniques will be applied to large administrative data sources, specifically MarketScan and the Medicare 20% sample. Aim 2 will harness rich data from deeply phenotyped cohorts to understand behavioral characteristics that influence the effectiveness of medications, and glean added understanding of the physiologic mechanisms through which pharmacotherapies act. These analyses will use data from the Atherosclerosis Risk in Communities (ARIC) study and the Multi-Ethnic Study of Atherosclerosis (MESA). Findings from the proposed Aims will advance understanding of the role of specific pharmacologic treatment strategies on CVD risk, and have potential to directly impact clinical practice, reinforcing or leading to reconsideration of existing clinical guidelines. This proposal is structured to allow trainees the flexibility to pursue their own hypotheses, motivated by their clinical experiences and interests. With support from this K24, Dr. Lutsey plans to expand the breadth of her POR research activities, recruit new junior clinical scholars into POR, and prepare them to make major scientific advances and launch their own independent research careers.

NIH Research Projects · FY 2024 · 2021-07

Project Summary Abdominal aortic aneurysm (AAA) is an important cause of morbidity and mortality in older adults. AAA rupture carries a ≥ 65% mortality rate. There are no direct pharmaceutical treatments for AAA, so the main management options are screening, secondary risk factor intervention, and surgical repair for large AAAs, which carries risk. Our previously funded AAA R01 “Identifying Epidemiological Risk Factors for Abdominal Aortic Aneurysm” established one of the few large population-based prospective US cohorts to identify etiologic risk factors for incident AAA. Among 15,792 ARIC participants followed for more than two decades, we ascertained 665 AAAs, identified novel middle-age risk markers for AAA, and estimated the lifetime risk for AAA from age 45 to be 5.6%. Building upon our original R01 and an ongoing proteomic project in ARIC, we propose a 4-year study to identify proteomics risk markers and investigate novel mechanisms and etiological pathways for AAA. Our specific aims are to: (1) Leverage a large panel of aptamer-based, plasma proteomics data (n=4,931 human proteins) in the entire ARIC cohort from Visits 2 and 3, to conduct a prospective study of proteomic risk markers for AAA (n=552 cases) over 24 years of follow-up, and to replicate significant proteins identified in nested AAA case (n=518)-cohort (n=833) samples from the prospective, population-based HUNT3 and SCCS studies. We will use a combination of targeted and agnostic approaches. To ensure the accuracy and generalizability of our findings, we will also identify commercial assays or develop targeted quantitative liquid chromatography-mass spectrometry assays for the top 5 novel, replicated, aptameric-based proteins and then compare the targeted protein levels with the aptamer-based measurements in 200 ARIC plasma samples. (2) Conduct genome-wide association study (GWAS) in ARIC for proteins identified and replicated in Aim 1, and replicate any significant genetic associations in HUNT3 (n=4,230), MESA (n=5,351), and published protein GWAS database. We will also conduct a Mendelian randomization study, incorporating data from the international AAA GWAS Consortium (10,204 AAAs and 107,766 controls), to elucidate the causal relation between significant protein biomarkers and AAA, followed by a network analysis to integrate the genomic and proteomic findings. This study will use unsurpassed proteomic resource in ARIC and other cohorts to identify new risk factors and mediators of AAA, with potential implications for AAA prevention and treatment.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY/ABSTRACT Since the World Health Organization declared COVID-19 a pandemic in March 2020, increasing evidence has shown that the disease affects multiple organs, including the central nervous system (CNS). Effects of COVID-19 on the CNS in the acute phase were documented clinically, by magnetic resonance imaging (MRI) and spectroscopy (MRS), by plasma biomarkers and at autopsy, with neurological symptoms manifesting in 1/3 to 2/3 of hospitalized, severe cases. After the acute phase, approximately 10% of patients experience prolonged illness, during which neurological symptoms (headaches, cognitive blunting, and fatigue) are among the top 10 symptoms reported by COVID-19 survivors. The underlying biology of these prolonged symptoms is unknown; therefore, prospective studies to systematically investigate the pathophysiology of such sequelae are urgently needed. Based on the clinical presentation of COVID-19, reports of COVID-related symptoms in the months following the infection, including reports of Parkinsonism and other delayed neurological and neurocognitive complications ranging from mild-to-severe, and known peripheral triggers of cerebral pathology, neuroinflammation (Aim 1), neurodegeneration (Aim 2) and cerebrovascular disease (CVD) (Aim 3) are expected to be important components of long-term CNS pathophysiology. The COVID BRain Advanced Imaging Network (COVID-BRAIN) was formed as a Consortium of six institutions to systematically and prospectively elucidate the long-term CNS pathophysiology of COVID-19 using highly sensitive, harmonized, advanced MRI/MRS technology at 3 tesla in conjunction with standardized neurological and neuropsychological evaluation and inflammatory blood biomarkers. Five sites that currently partner in other multi-site neuroimaging initiatives (University of Minnesota, Mayo Clinic Rochester, Harvard University/Massachusetts General Hospital, Johns Hopkins University, Houston Methodist Research Institute) will collect longitudinal multi-modal MRI (T1, FLAIR, diffusion MRI, susceptibility-weighted MRI, single- and multi-voxel MRS and pseudo-continuous arterial spin labeling), clinical, neurocognitive and blood biomarker data from laboratory confirmed post-COVID cases with neurological symptoms (N=200) and matched controls (N=100). The Laboratory of Neuro Imaging (LONI) at the University of Southern California will serve as the data management site. Group differences and change over time in MR markers indicative of neuroinflammation, neurodegeneration, hypoxia/ischemia and CVD and their associations with specific neurological symptoms, cognitive function, and inflammatory blood biomarkers will be investigated. The mechanistic insights provided by this study will inform the care and treatment of patients that are expected to suffer long-term consequences of the pandemic for the years to come.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract Memory CD8 T cells with varying functional characteristics are generated after infection or immunization. We previously defined a population of T cells within the CD62Llo effector memory compartment that express high levels of effector molecules and continue to persist into the memory phase. Although over time in specific pathogen free mice LLECs tend to wane in number, they represent a substantial fraction of CD8 memory T cells in ‘dirty mice’ and dominate the secondary and tertiary memory pools. Importantly, our prior work showed that these ‘long-lived effector cells’ (LLEC) are the most robust memory T cells for mediating antigen-specific clearance of systemic viral and bacterial pathogens. Our recent RNA sequencing data indicates that LLEC may achieve this through unique expression of multiple NK cell-associated receptors as well as chemokine and trafficking molecules that may enforce their strict localization to the vasculature at the steady state. In this proposal we will determine: 1) if LLECs participate in tissue-initiated infections through either extravasation or from their position within the vasculature, 2) if NK cell receptors modulate LLEC function, and 3) if the LLEC subset uniquely thrives during inflammation for its persistence. Our proposal leverages recent transcriptional analysis along with innovative mouse models and technical approaches to understand the signals governing how circulating memory T cell populations mediate protective immunity. We predict our studies will expose novel considerations for generating robust memory T cell function, ultimately leading to improved vaccination and immunotherapy approaches.

NIH Research Projects · FY 2026 · 2021-07

ABSTRACT Executive function (EF) improves dramatically during childhood and adolescence, and failures of EF are associated with both a broad range of negative outcomes and diverse mental illnesses. The brain circuits responsible for EF are spatially distributed, and include the fronto-parietal, cingulo-opercular, and salience systems. These networks have typically been studied using standardized network atlases, which assume a straightforward mapping between structural and functional neuroanatomy across individuals. However, multiple independent efforts in adults using precision functional mapping techniques have recently demonstrated that there is marked inter-individual variation in functional topography, which is defined as the spatial distribution of functional networks on the cortex. The over-arching hypothesis of this proposal is that individual variation in functional topography is a critical determinant of EF in youth. Our collaborative team recently published the first report of individualized functional networks in children using a cross-sectional sample (Cui et al., Neuron 2020). In this proposal, we will build upon this initial work by replicating and generalizing this finding using two large cross-sectional datasets with high-resolution imaging: the Healthy Brain Network (HBN; n=5,000) and Human Connectome Project: Development (HCP-D, n=1,300). Critically, we will also leverage the unprecedented resources of the Adolescent Brain and Cognitive Development Study (ABCD, n=11,572) to delineate within-subject change in personalized networks. In this proposal, we will first harmonize these massive data resources using advanced techniques originally developed for statistical genomics (Aim 1). Next, we will describe how personalized networks evolve with age (Aim 2) and predict EF (Aim 3). Finally, we will use machine learning tools to discover how the functional topography of personalized executive networks predict dimensions of psychopathology in a data-driven manner (Exploratory Aim 4). Throughout, we will adhere to best practices of open science to maximize reproducibility, and ensure that all processed data, code, and results are openly shared with the neuroscience community. Together, this research will establish that functional topography is essential for understanding EF, and will motivate trials of personalized neuromodulatory therapies.

NIH Research Projects · FY 2025 · 2021-07

7. PROJECT SUMMARY / ABSTRACT Periodontitis is a chronic inflammatory condition characterized by the destruction of the periodontium and is the leading cause of tooth loss in adults. It is driven by a dysbiotic microbial biofilm that colonizes the gingival sulcus. We seek to identify and characterize the local immune response to the microbial biofilm that leads to periodontitis. Recently, CD69 engagement on T regulatory cells was reported to induce immunosuppressive activities. A natural ligand for CD69-mediated activation of Tregs is calprotectin (CLP; S100A8 complexed to S100A9; S100A8/A9; MRP8/14). When expressed in stratified squamous epithelia, this divalent cation-binding complex appears to contribute to intraepithelial antimicrobial defense. When released from infected or desquamating keratinocytes or neutrophils, however, CLP may interact with CD69+ T regulatory or T helper 17 cells, ultimately suppressing the immune response. If so, CLP may function during the initiation of periodontitis contrary to its postulated function as a proinflammatory “alarmin”. Using a global CLP null mouse, our preliminary data suggest that the net effect of CLP dampens the recruitment of an acute inflammatory infiltrate and limits periodontal bone destruction in a ligature-induced experimental periodontitis model. We will now explore a modified mouse model of ligature-induced periodontitis primed with Porphyromonas gingivalis (Pg). and in the resolution phase We hypothesize that CLP signals through CD69 during the initiation and resolution phases of experimental periodontal inflammation to dampen the destructive cellular infiltrate in the gingiva. To test our hypothesis, we will: 1: Characterize the differences in the inflammatory cell infiltrate attributable to CLP during the initial and resolution phases of experimental periodontitis. 2: Determine how Pg-primed Treg cells modulate the recruitment of the initial inflammatory cell infiltrates through CD69 signaling and CLP. 3: Determine the contribution of Pg-primed Th17 cells to modulating recruitment of the initial and resolution phase and resolution phase inflammatory cell infiltrates attributable to CD69 signaling and CLP. To our knowledge, we are the first group with data suggesting that CLP dampens the innate immune response in a CD69-dependent manner. We have the tools to explain how CLP contributes to recruitment of innate immune cells by affecting Treg and Th17 cells in vivo using a murine model of periodontitis. Ultimately, we will characterize how CLP and CD69 signaling in Treg and Th17 cells shapes the immune cell environment in the gingiva to drives either protection of periodontal tissues or destruction of alveolar bone. The results obtained here will be used to design therapeutic interventions directed at boosting or inhibiting the activity of CLP. Critical steps will be identified that might be amenable to targeted therapeutic intervention in humans. Ultimately we aim to reduce the economic and personal burden of periodontal diseases.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract Preeclampsia is a complex disease of pregnancy involving cardiovascular, immune, and placental dysfunction. It is the leading cause of maternal-fetal morbidity and mortality. Preventative options are poor and the only effective treatment is the delivery of the often pre-term fetus and the dysfunctional placenta. The current proposal aims to investigate the potential of personalized medicine in the form of cellular immunotherapy as a novel preventative measure for the prevention of preeclampsia. Elevated secretion of arginine vasopressin precedes the development of physiological symptoms of preeclampsia in humans, and infusion of vasopressin throughout gestation into wild-type mice is sufficient to initiate the cardiovascular, renal, and immune dysregulation observed in human preeclampsia. Although vascular and renal complications are observed clinically in preeclampsia, immune dysfunction is central to the pathogenesis of preeclampsia. Regulatory dendritic cells have been shown to induce immune tolerance and are a current cellular immunotherapy in ongoing clinical trials for several other immune mediated diseases. The proposed study will test the hypothesize that treatment with regulatory dendritic cells will prevent the cardiovascular, renal, and immune features of preeclampsia. The overall objective of this proposal is to determine if DCreg administration is a potential preventative for preeclampsia. In the following aims, utilizing the chronic AVP infusion mouse model of preeclampsia and primary human mononuclear cells, we will investigate 1) the potential of DCreg to prevent the cardiovascular, renal, and immune features of preeclampsia in mice and 2) the ability of human mononuclear cells obtained from women with chronic hypertension and/or preeclampsia to differentiate into functional DCreg. The vision for my career is to develop an independent, NIH-funded research program that transforms women’s cardiovascular healthcare through innovative scientific discovery and the education of future scientists. To continue to grow as an independent investigator and to achieve my career goals, the objective of the career plan is to develop skills in the following areas: 1) training in assessment of vascular function as it relates to models of preeclampsia; and 2) developing skills in translational clinical research. These will be accomplished through both didactic coursework and experiential training through direct interactions with my mentors and colleagues. If regulatory dendritic cells prevent preeclampsia in the present study, this data will identify a novel cellular immunotherapy for preventing preeclampsia. Future studies would be aimed at further understanding the mechanism(s) of regulatory dendritic cell action and the potential of regulatory dendritic cells in preventing preeclampsia. This project is innovative as it proposes a powerful, novel treatment as a possible prevention for preeclampsia: personalized cellular immunotherapy.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY/ABSTRACT Natural killer (NK) cells are innate lymphocytes that mediate cellular cytotoxicity against virally infected and malignant cells. Upon activation, they also release chemokines and cytokines that help prime and coordinate the adaptive immune response. Because of these functional properties, there is continued interest in developing NK cell products for immunotherapy. However, this effort is hindered by our limited understanding of human NK cell differentiation and the transcription factor networks that regulate this process. In work supported by current K99/R00 funding, we generated a considerable amount of preliminary data comprehensively characterizing the transcriptional and epigenetic landscapes in sorted NK and CD8+ T cell subsets isolated from cytomegalovirus (CMV) seropositive donors. To complement these analyses, we performed flow cytometry-based analyses of transcription factor expression and ChIP-seq on sorted peripheral blood NK cell subsets to construct transcription factor networks that potentially regulate key steps in NK cell differentiation. The culmination of this work revealed a central role for Bcl11b. This was completely unexpected given several detailed studies in mice showing that Bcl11b expression is restricted to the T, NKT, and ILC2 lineages and acts to enforce T cell identity. Our preliminary results also revealed a reciprocal relationship between Runx2 and Bcl11b, with Runx2 regulating a set of transcription factors enriched in immature NK cells and Bcl11b regulating genes enriched throughout canonical NK cell maturation and in adaptive NK cells exhibiting a T cell-like gene expression signature. In this application we will test the hypothesis that Runx2 is a major hub that enforces the epigenetic identity of immature CD56bright NK cells, while Bcl11b suppresses the Runx2-directed transcriptional program to drive canonical NK cell differentiation, maturation, and cytotoxic effector function. We also posit that high levels of Bcl11b contribute to the T cell-like features of CMV-induced adaptive NK cells. We propose two independent aims to test our hypotheses. In the first aim, we will use overexpression and knockdown strategies in sorted NK cell subsets to define the roles of Bcl11b and Runx2 during canonical NK cell differentiation. These results will guide subsequent experiments designed to determine whether Bcl11b overexpression in hematopoietic progenitor cells (HPCs) promotes NK cell maturation and effector function both in vitro and after adoptive transfer into mice with established tumors. In the second aim, we will determine the role of Bcl11b in promoting T cell-associated features of CMV-induced adaptive NK cells and test the hypothesis that high levels of Bcl11b along with activating receptor and cytokine receptor stimulation drives adaptive NK cell differentiation and maturation. We will also test the hypothesis that adaptive NK cells mediate superior antitumor function in vivo relative to canonical NK cells. Results generated from this proposal will define the role of Bcl11b in promoting canonical and adaptive NK cell differentiation, determine whether Bcl11b overexpression enhances NK cell-mediated antitumor activity, and definitively test the antitumor function of adaptive NK cells.

NIH Research Projects · FY 2025 · 2021-07

PROJECT SUMMARY Genetics is a foundational discipline central to biomedical research and clinical practice. Our mission is to train geneticists for the 21st century. To achieve this goal, we have designed and implemented an interdisciplinary training program to transform graduate education in genetics and genomics at the University of Minnesota. Our Specific Aims are: (i) to provide students with rigorous and quantitative training in the fundamentals and state- of-the-art advances in genetics, genomics, and computational biology; (ii) to leverage the many collaborative research and educational strengths of our institution to provide new interdisciplinary student-centered graduate training opportunities that transcend departmental and collegiate boundaries; and (iii) to generate a diverse cohort of well-trained scientists able to integrate their broad skillset and their hunger for lifelong learning into their work across a multitude of careers in the biomedical workforce. This program will enable trainees to pursue many impactful career opportunities in academia, biotechnology, government, health care, or education created by recent advancements in genetics and genomics, including careers in fields that are undergoing rapid change, as well as careers in areas that do not presently exist or that are nascent. In recognition of the fact that the geneticists of tomorrow must possess quantitative acumen, we have not only developed new courses emphasizing bioinformatics and computational approaches, but we have also integrated these into our training program through a major redesign of the graduate curriculum pursuant to evaluation by internal and external review panels. In a sense, genetics and genomics are foundational fields that transcend and unify multiple disciplines within the biological sciences because what sets living things apart from inanimate objects is that all living things possess within them molecular blueprints for their construction, genomes. An impactful aspect of this academic and research training program is that it seeks to transcend traditional departmental boundaries by bringing together researchers from multiple disciplines within the biomedical sciences who utilize genetic and genomic analyses in their research. In so doing, this training program seeks to promote interdisciplinary research, foster collaborative team science, and implement individualized student-centered educational and career development programs. Our training program includes 50 highly productive and collaborative faculty from ten departments across five colleges at the University of Minnesota. We are seeking support for six PhD trainees for two years each beginning in year two of their academic program, with funding for year one provided by institutional funds. In addition to their thesis research, trainees will participate in professional development programs aimed at promoting their transition into and long-term success in biomedical science-related careers.

NIH Research Projects · FY 2024 · 2021-07

Summary: Distribution of dietary vitamin A/all-trans retinol/ROL throughout the body is critical to maintain retinoid function in peripheral tissues and to ensure optimal vision. In humans, dietary vitamin A is absorbed in the small intestine, stored in the liver, and secreted into circulation bound to serum retinol binding protein 4 (RBP4-ROL). A receptor-mediated system for systemic RBP4-ROL uptake, storage and transport is essential not only to ensure availability for optimal ocular retinoid signaling for photoreceptor function, but also to prevent cellular toxicity associated with excessive retinoid accumulation. STRA6, the only known receptor for circulatory ROL bound RBP4 in the eye, is not expressed in systemic tissues proposed to facilitate the uptake of RBP4- ROL. This indicates the existence of additional vitamin A transporters in such tissues. The objectives of this proposal are 1] to determine the physiological role a novel vitamin A transporter, the retinol binding protein 4 receptor 2 (RBPR2) in facilitating the systemic uptake of dietary ROL bound RBP4 for vision, and 2] to investigate if modulation of such eye related ROL transporters could limit substrate availability required for toxic retinoid biogenesis and thus improve vision in patients with inherited retinal degenerative diseases. The long-term goal is to identify the physiological mechanisms of RBPR2 for RBP4 binding and ROL transport in retinal health and disease states. The central hypothesis is that RBPR2 has high affinity binding for RBP4-ROL in tissues devoid of STRA6 and that its physiological function is critical to ensure and regulate dietary vitamin A uptake and delivery to the eye in the support of vision. The rationale underlying this proposal is that completion will fill the knowledge gap of how dietary vitamin A is sequestered into systemic tissues from RBP4, transported and stored in peripheral tissues lacking STRA6, for eventual distribution to the eye for vision. The central hypothesis will be tested by pursuing three specific aims that will in Specific Aim 1: Determine the functionality of RBPR2 for vitamin A transport; Specific Aim 2: Determine the physiological role of RBPR2 for systemic RBP4-ROL transport in vision; and Specific Aim 3: Determine if modulation of Rbpr2 activity attenuates inherited retinal degenerative diseases. We will pursue these aims using an innovative combination of structural analysis, biochemistry, cell biology, physiology and novel animal models aimed at exploring the in vivo requirements of RBPR2 for ROL transport for photoreceptor health, vision and in retinal disease. The proposed research is significant because it will determine for the first time the mechanisms influencing circulatory RBP4-ROL uptake, storage and transport into the eye, and explore strategies aimed at modulation of such eye related ROL- transporters for improving vision in humans with inherited retinal diseases. The proximate outcome of the proposed research will provide information on a novel ROL transporter, RBPR2, that will improve understanding of human disease states, particularly blindness, associated with impaired blood vitamin A homeostasis or ocular vitamin A excess and could yield concepts for their prevention and therapy.

NIH Research Projects · FY 2025 · 2021-07

Neighborhood parks hold great potential as attractive and accessible settings to promote and enhance physical activity (PA) among racially/ethnically diverse low-income youth. However, little park-based intervention research has been conducted to explore how this potential might be realized. No previous park-level intervention has experimentally evaluated youth sports program fee waivers or intensive park outreach strategies to increase child enrollment and participation in park-based youth sports programs. The proposed research combines these promising intervention strategies into a single intervention and uses a gold-standard randomized controlled study design, and focuses on a low-income, racially/ethnically diverse sample at highest risk for developmental declines in physical activity. The proposed study evaluates the effects of a park-level intervention to increase moderate/vigorous physical activity (MVPA), and decrease sedentary time among low- income racially/ethnically diverse children ages 6-12 yrs. Neighborhood parks (n=24) will be randomized for a 2-year period to one of two experimental conditions: 1) Intensive Outreach and Youth Sports Program Fee Waiver; or 2) Comparison Condition: Current Outreach and Youth Sports Program Fee Structure. The intervention outreach and fee waiver components are expected to support parents to enroll their child in park- based youth sports programs by decreasing social/cultural and economic barriers to child enrollment and participation. The evaluation cohort will consist of 432 children ages 6-12 yrs who reside in one of the 24 park neighborhoods randomized to the experimental conditions. Measurements from individual children will be collected at baseline, 6, 12, and 24 months. Specific Aims of this research are: Specific Aim 1: To evaluate the effects of a park-level intervention on changes in child MVPA over 6, 12 and 24 months; Specific Aim 2: To evaluate the effects of a park-level interventions on changes in child sedentary behavior over 6, 12 and 24 months; Exploratory Aim 3: To evaluate the effects of a park-level interventions on changes in parent light activity and sedentary behavior over 6, 12 and 24 months. Hypothesis 1: Children who live in a neighborhood where the park is randomized to intensive park outreach and fee waiver will have larger increases in MVPA and decreases in sedentary behavior at 6, 12 and 24 months relative to children who live in a comparison neighborhood. Hypothesis 2: Parents of children who live in a neighborhood where the park is randomized to intensive park outreach and fee waiver will have larger increases in light PA and decreases in sedentary behavior at 6, 12 and 24 months relative to parents of children who live in a comparison neighborhood. IMPACT: The proposed research will provide evidence about the effectiveness of a park-based intervention that includes intensive park outreach and youth sports program fee waivers on changes in child physical activity in low-income, diverse urban communities. It will provide critical information needed to support policies and programs that reduce disparities in physical activity among high risk youth.

NIH Research Projects · FY 2025 · 2021-07

This K24 grant will provide the PI, Dr. Lin Yee Chen, a midcareer NIH-funded patient-oriented research (POR) investigator, with the protected time and support needed to (1) accelerate his current trajectory in mentoring junior clinicians and investigators who are conducting POR in cardiovascular (CV) science, (2) acquire additional training in mentoring methods, machine learning, and omics science, (3) promote his current research that aims to define the mechanisms underlying the relationship of the abnormal atrial substrate—atrial cardiomyopathy—and atrial fibrillation (AF) to ischemic stroke and other CV outcomes. Trainees in his AF Clinical Research Group will be recruited from the NIH-funded T32 training programs in the University of Minnesota's Division of Cardiology, Division of Epidemiology, and Division of Biostatistics; Department of Medicine Physician Scientist Training Program; KL2 and TL1 Programs of the CTSI; and graduate programs (MS in Clinical Research, MPH, and PhD) in the School of Public Health. For his career development, Dr. Chen will hone his mentoring skills and learn new skills in cutting-edge areas (machine learning and omics science) through focused study, selected coursework, seminars, and guidance from senior collaborators with domain expertise. Finally, this grant will support a research project that is based on the Atherosclerosis Risk in Communities (ARIC) Study, which extends Dr. Chen’s ongoing work to elucidate the role of atrial cardiomyopathy in driving AF-related outcomes. The specific aims are: (1) Identify atrial cardiomyopathy subtypes by using machine learning approaches to analyze the extensive data at ARIC Visit 5 (2011-13): clinical, 2D-echocardiographic, heart rate variability, arterial stiffness, ECG, and multi-omics data, (2) Evaluate association of subtypes with ischemic stroke and other CV outcomes, and (3) Discover specific risk factors for subtypes by analyzing risk factor measures collected at Visits 1-4 (1987-98). Crucially, findings will be validated in 2 independent community-based cohorts: Cardiovascular Health Study (CHS) and Multi-Ethnic Study of Atherosclerosis (MESA). This project has significant impact: (1) This K24 grant will provide the PI with the protected time to enhance his current mentoring of trainees involved in POR. By assembling a team of senior collaborators that comprise experts in mentoring, data science, and molecular epidemiology, the PI provides an outstanding platform for his mentees to acquire cutting-edge skills in POR, (2) By resolving heterogeneity in atrial cardiomyopathy, the team will advance the NIH’s Precision Medicine initiative by personalizing current treatment for patients based on biological underpinnings, (3) The findings will lead to a clinically meaningful improvement in classification of stroke risk in patients with AF, which will improve patient outcomes, (4) By efficiently leveraging existing resources of deeply phenotyped NHLBI cohorts, this project will fill critical knowledge gaps in prevention and treatment, thus achieving a sustained and powerful impact on CV public health, clinical practice, and education of the next generation of researchers in POR.

NIH Research Projects · FY 2025 · 2021-06

SUMMARY Type 1 diabetes (T1D) is an autoimmune disease resulting from a breakdown in immunological tolerance caused by T cell-mediated destruction of islet beta cells. Diabetes is orchestrated by HLAII- restricted CD4+ T cells, through cellular interactions with both B cells and CD8+ T cells, resulting in autoantibody production and beta cell death, respectively. While anti-islet autoantibodies are currently the best predictors of T1D development, screening is limited to four islet antigens and no T cell biomarkers exist. Despite years of research, it is still unclear which antigen-specific CD4+ T cells initiate T1D. New evidence suggests that hybrid peptides (HP) formed from the fusion of islet β cell proteins may be critical antigens in T1D as recent studies identified HP-reactive CD4+ T cells from T1D patients and diabetic mice in vitro. These neo-antigens escape central tolerance and must be controlled by peripheral mechanisms including anergy or regulatory T cell control. In preliminary studies, we identified HP-specific CD4+ T cells in diabetic mouse models using tetramer reagents, and showed they are pathogenic and cause T1D in mouse transfer models. More importantly, we can block spontaneous T1D in the NOD mouse model targeting one hybrid peptide when presented in mouse MHCII using peptide-specific:MHCII blocking antibodies. Thus, we hypothesize that HPs are critical antigens and that autoreactivity to HPs initiates T1D. The goals of this proposal are to determine if HP-specific CD4+ T cells initiate T1D and if targeting them can lead to tolerance as a prevention or cure for T1D. The second goal of the grant is to determine if HP specific cells are relevant for human T1D and use of scRNA-seq analysis to uncover critical clues about shared transcriptional programs related to the pathogenic potential between human and mouse HP reactive T cells. Completion of this project could lead to better biomarkers to predict T1D risk and disease progression.

NIH Research Projects · FY 2025 · 2021-06

SUMMARY/ABSTRACT Somali women living in the U.S. have lower cervical cancer screening rates than the U.S. general female population. This disparity is due to a range of factors, including limited awareness of HPV and cervical cancer, cultural and religious beliefs, mistrust of health care providers, concerns around modesty and circumcision, and low provider self-efficacy to perform Pap tests on circumcised women. HPV self-sampling is an emerging cervical cancer screening modality that may address common screening barriers among Somali women, particularly those related to modesty and circumcision. HPV self-sampling is accurate for detecting pre- cancerous cervical lesions and effective in reaching underscreened women. In addition, our pilot work has demonstrated the feasibility and acceptability of HPV self-sampling in Somali women. While research has focused primarily on home-based HPV self-sampling, there is an untapped opportunity to offer HPV self- sampling in the primary care setting. Offering HPV self-sampling in primary care could effectively increase cervical cancer screening rates in Somali women by positioning providers to address screening barriers, enabling clinics to opportunistically fit in HPV self-sampling with other appointments, and providing an alternative modality for circumcised women. We propose a Hybrid Type 2 effectiveness-implementation design to assess the effectiveness and implementation of a patient-centered, culturally-tailored HPV self-sampling intervention for Somali women. Guided by the Consolidated Framework for Implementation Science and Social Cognitive Theory, we will conduct focus groups with Somali patients and interviews with providers to identify patient-, provider-, clinic-, and systems-level factors to inform refinement of intervention materials and development of implementation strategies (Aim 1). After tailoring the intervention, we will then implement HPV self-sampling in 2 primary care clinics, and evaluate changes in Somali women’s cervical cancer screening rates one-year pre and one-year post implementation (Aim 2). Changes will be compared with Somali women attending 27 control clinics followed over the same time period, using difference-in-difference methods. Finally, using RE-AIM, we will conduct a post-implementation mixed methods analysis of the processes and strategies needed to successfully implement HPV self-sampling in primary care for Somali patients (Aim 3), including interviews with providers and Somali women. Our hypothesis is that implementing HPV self-sampling in primary care will lead to increased uptake of cervical cancer screening in Somali women and that an implementation science based analysis of the processes needed to successfully implement the intervention will lead to sustainable, novel strategies to support the sustained integration of HPV self-sampling into primary care.

NIH Research Projects · FY 2025 · 2021-06

The last decade ushered in amazing advances in neuroimaging due to transformative developments in magnetic resonance and optical imaging techniques as well as in computational and modeling tools. A common thread in these advances is their multi-disciplinary nature, requiring collaborations among medical researchers, engineers, physicists, mathematicians, and data scientists, among many others. In order to continue the pace of technical advances in neuroimaging and to exploit their unique capabilities for brain research and medical applications, it is critical to train the next generation of neuroimaging specialists in a setting 1) with an abundance of state-of-the-art tools, 2) with a programmatic interest in developing novel neuroimaging technologies and biomedical applications, and 3) where trainees can carry out groundbreaking research under multi-disciplinary mentorship. The University of Minnesota (UMN) has an excellent tradition of training neuroimaging postdocs in its Medical School and its College of Science & Engineering. It is also the home of major, world-renowned efforts in neuroimaging technology development and novel biomedical applications of neuroimaging. The proposed Minnesota Neuroimaging Postdoctoral Training Grant aims to give 14 postdoctoral fellows multi-disciplinary skills in neuroimaging technology development and advanced biomedical applications, guidance in career development, and social and networking support through intense two-year neuroimaging fellowships at UMN. Each fellow's primary research will be conducted on a multi-disciplinary project that combines their background with another field with the express goal of developing new neuroimaging technologies. Each fellow will be co-mentored by two faculty selected from the 40+ participating faculty in this grant: one that will directly supervise the research project, and one that represents a core area related to the research project. Fellows will take at least two courses to broaden their skillset and prepare for either an academic or industry research career. They will participate in an Annual Retreat and twice-monthly seminars that will cover research and career development topics such as responsible conduct of research, scientific rigor and reproducibility, grant writing, and other key subjects. They will also participate in UMN's numerous neuroscience conferences, symposia, and workshops, along with well-established UMN outreach programs to high schoolers and undergraduates interested in STEM fields. The program will be managed by an Executive Board. Management plans include a rigorous, ongoing evaluation process that incorporates an external Advisory Board and the University of Minnesota's internal research and assessment services.