University Of Minnesota

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY/ABSTRACT The overarching objective of this trial is to test the impact of a full-service mobile food market (“a grocery store on wheels”) on diet quality, food insecurity (i.e., uncertain or not enough access to food for a healthy life) and food purchases in communities experiencing low food access and low incomes. We will also explore personal, social, behavioral, and environmental factors that may influence adoption of mobile market shopping. The long-term goal of our work is to improve diet/weight-related health outcomes by increasing access to and intake of healthy foods for individuals who are more likely to be impacted by poor nutrition and diet/weight-related health outcomes, such as type 2 diabetes and high blood pressure. Lack of affordable, healthy food access and food insecurity are believed to be important contributing factors to these outcomes. Mobile food markets have been proposed as a strategy for improving diet-related health outcomes because they aim to improve access to low-cost, healthy food. Mobile food markets are increasingly available nationally and have been supported by state legislation, but little is known about their effectiveness. Full-service mobile markets may improve multiple aspects of the diet by providing foods to meet all dietary needs through a convenient one-stop shop. The full-service mobile market to be tested in this proposal (Twin Cities Mobile Market) sells staple foods from a bus that regularly visits low-income neighborhoods. Foods are sold at prices 10% below those of grocery stores. SNAP/EBT is accepted, and a state-funded fruit/vegetable incentive program (Market Bucks) is available to shoppers. Working in partnership with our community team members, we will enroll 6 community sites (clusters) in two waves (12 total sites/clusters) and recruit 20 participants per site (N=240). We will collect baseline data and randomize sites to either receive the full-service mobile market intervention or serve as the waitlist control. We will then implement the full-service mobile market at intervention sites, follow participants for one year, and collect follow-up data. After follow-up data collection, waitlist control sites will receive the full-service mobile market intervention. For Aim 1, changes in diet quality (our primary outcome measured with the Health Eating Index-2015) and food insecurity (measured with the 18-item U.S. Adult Food Security Survey Module) will be assessed. For Aim 2, changes in food purchases will be objectively measured by collecting one month of food purchase receipts at baseline and follow-up data collection. We will also assess mobile market food purchases during the implementation period with customer loyalty cards. For Aim 3, we will explore the factors that influence adoption of mobile market shopping. Findings will provide evidence on the effectiveness of a full-service mobile market to improve diet, food security, and food purchasing outcomes. This innovative research will provide timely evidence to inform mobile market sustainability, policy, and legislative decisions.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY More than 1.5 billion people are infected with helminths worldwide, predominantly distributed in tropical and subtropical areas. On the other hand, in developed countries with markedly reduced infectious diseases, there is a continuing increase in the incidences of allergic, inflammatory, and autoimmune diseases. The hygiene hypothesis proposes that an under-stimulated immune system resulting from the absence of exposure to helminths, predisposes to autoimmune and allergic inflammation. There is an urgent need for new preventive and therapeutic medicines for mucosal infection and inflammation. The research in my laboratory has been focused on the pathophysiological role of O-linked N-Acetylglucosamine (O-GlcNAc) modification on intracellular proteins at serine and threonine residues. The long-term goal is to elucidate the regulatory mechanisms and physiological functions of O-GlcNAc signaling in intestinal homeostasis and mucosal host defense. In the proposed study, we will test the hypothesis that O-GlcNAc transferase (OGT), by activating STAT6 signaling, promotes the differentiation of IL-25-producing tuft cells and facilitates IL-33 secretion from goblet cells, thus evoking type 2 immune responses for tissue repair and inflammation control. In Aim 1, we seek to define the functional impact, upstream activating signals, and downstream targets of STAT6 O-GlcNAcylation in tuft cell differentiation, type 2 immune activation, and helminth expulsion. In Aim 2, we identify a novel, common target of OGT and STAT6 that colocalizes with IL-33 in goblet cells and mediates the unconventional secretion of IL- 33 to initiate type 2 immune responses. In Aim 3, using pharmacological and genetic approaches to increase global protein O-GlcNAcylation in the intestinal epithelium, we expect to establish that the OGT-STAT6 pathway is required for intestinal homeostasis and mediates the therapeutic effect of helminths in colitis. The proposed study will provide valuable insights into the development of new intervention strategies to eradicate parasitic worms in areas of poverty in the developing world and to treat inflammatory bowel disease in industrialized countries.

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY / ABSTRACT Metastatic relapse may occur in patients with ER+ breast cancer decades after original diagnosis. Most breast cancer related deaths are caused by metastasis and thus identifying at-risk patients and developing therapies to prevent reactivation are a crucial challenge. These efforts have been impeded by a lack of understanding of cancer cell dormancy in the bone marrow, believed to be the cellular source of metastatic relapse. It is not well understood how tumor cells interact with the bone marrow microenvironment and how these interactions regulate tumor cell dormancy and escape. My proposed research seeks to develop a mouse model of dormancy and investigate the spatial organization of the bone marrow niche in patient samples in the mentored K99 phase and develop a biomaterial model of dormancy in the independent R00 phase. I hypothesize (i) cancer stem cells are a subset of disseminated tumor cells responsible for metastatic relapse and (ii) the bone marrow niche, including the healthy stem cell niche, facilitates dormancy and reactivation of these cells. In Aim 1, I will develop a novel mouse model of hormone responsive breast cancer dormancy in bone marrow and evaluate the presence, phenotype, and microenvironmental regulation of disseminated tumor cells using optical tissue clearing/3D imaging of whole bone. In Aim 2, I will investigate the hypothesis that tumor cell interactions with the bone marrow niche control tumor cell phenotype via high dimensional spatial analysis of bone marrow biopsies from patients with breast cancer including imaging mass cytometry (IMC) and spatial single cell RNA sequencing (scRNAseq). In Aim 3, I will develop a biomaterial model of the dormant bone marrow niche via creating mechanical mimics of the three distinct compartments of bone marrow and evaluate the role of mechanosensing in induction and maintenance of dormancy. In the K99 phase of the award, Prof Max Wicha will serve as my main mentor. Dr. Wicha is a pioneer in the cancer stem cell field and is an expert in breast cancer biology and metastasis. I will work with and consult my collaborators and mentoring team, including Prof Fei Wen (imaging mass cytometry), Evan Keller (single cell spatial analysis program), Dr. Dafydd Thomas (pathology core), Prof Gary Luker (microscopy), Prof Monika Burness (breast cancer clinical oncology) and Prof Sofia Merjaver (breast cancer molecular biology). My K99 training will consist of developing a novel mouse model of bone marrow dormancy and learning bioinformatics approaches to analyze spatial contributions to cellular phenotype in IMC and scRNAseq data to propel me toward developing a synthetic dormant bone marrow niche mimic using biomaterials during the independent R00 phase. In sum, the proposed research will address an urgent, unmet need to identify the role of the bone marrow niche in breast cancer dormancy and reactivation, which may provide a path forward for identifying patients at higher risk of metastasis and developing therapies against reactivation.

NIH Research Projects · FY 2026 · 2022-02

Project Summary/Abstract The proposed R01 project will feature a robust collaboration between Lutheran Social Service of Minnesota (LSS-MN) and the University of Minnesota to evaluate the “Porchlight Project,” a novel adaption of its volunteer programs that serve older persons with Alzheimer’s disease or Alzheimer’s disease related dementia (AD/ADRD). The Porchlight Project is multicomponent training delivered to volunteers and includes three established online training modules on person-centered dementia care; a four-session online training program that demonstrates to volunteers how to apply person-centered dementia care knowledge to their interactions with persons with dementia and their caregivers; and ongoing monthly coaching sessions. We will evaluate the real-world efficacy of the Porchlight Project throughout Minnesota and randomly assign a minimum of 171 persons with AD/ADRD, family caregivers, and volunteers across 19 Regional Program Coordinator regions to one of two groups: one that receives the Porchlight Project over a 12-month period and a usual care control condition that receives standard volunteer support. An embedded experimental mixed methods design will be utilized that will incorporate various qualitative data collection elements within the 12-month randomized controlled evaluation of the Porchlight Project (Stage III of the NIH Stage Model). Twelve month outcomes that we hypothesize the Porchlight Project will influence include: 1) increased volunteer competence; 2) increases in quality of life and reduced loneliness for the person with AD/ADRD; and 3) increased self-efficacy, decreased stress, and improved well-being for caregivers. The Porchlight Project offers a potentially efficient, wide-ranging service model for states and communities to implement that can facilitate the “dementia capability” of their various volunteer programs. Moreover, creating effective linkages to community-based long-term services and supports addresses several goals of the National Plan to Address Alzheimer’s Disease as well as the recent Dementia Care Services and Support summits.

NIH Research Projects · FY 2025 · 2022-02

ABSTRACT Despites improvements in childhood cancer survival in the last several decades, marked racial, ethnic, and socioeconomic disparities in outcomes persist. Compared with non-Hispanic white children, non-Hispanic black and Hispanic children experience lower survival from many cancers, including leukemia, the most commonly diagnosed cancer in children. The underlying causes of these survival differences are poorly understood and may vary by cancer type, and both biological and socioeconomic pathways have been proposed. Recent evidence has suggested that lower socioeconomic status (SES) is associated with survival from some childhood cancers. The Children's Oncology Group (COG) is an international clinical trial cooperative group of over 200 hospitals which together treat more than 90% of all children and adolescents diagnosed with childhood cancer in the United States and Canada. In 2007 the COG opened the Childhood Cancer Registration Network (CCRN; COG protocol ACCRN07) to create a research registry. A total of over 56,000 childhood cancer cases were enrolled on ACCRN07 through the end of enrollment on December 8, 2017. All children and parents enrolled on ACCRN07 provided address information which was current at the time of diagnosis. We will work with investigators at the Minnesota Population Center to geocode all ACCRN07 patients with a valid U.S. address, contextualize with socioeconomic status data, and return small-area SES data to COG for dissemination. We will contextualize each geocoded address with Census data at the block level using seven variables from these data we will use factor analysis to derive a five-level SES indicator. We will then examine the influence of SES on risk of minimum residual disease at the end of induction therapy, relapse, other serious toxicities and adverse events, and survival in >9,500 acute lymphoblastic leukemia (ALL) patients. Over 9,500 ALL patients on ACCRN07 with a valid address will also have been treated on COG protocols. Ours will be the first study to evaluate SES as a predictor of childhood cancer outcomes on a large scale within the Children's Oncology Group, and will include detailed cytogenetic and molecular characterization of each tumor. Additionally, to our knowledge, this will be the first analysis of SES predictors of short-term treatment toxicities. We will create a highly useful resource on a large scale for a contemporary cohort of childhood cancer patients. Our findings will have translational potential in that outcomes related to SES may indicate the need to develop tailored interventions for low-resource patient populations. Additionally, this cohort's utility will extend beyond outcomes of therapy and into survivorship with linkages to the National Death Index (NDI) to obtain mortality data. Our long-term goal is to understand the factors that contribute to disparities in childhood cancer relapse, survival, and the burden of morbidity in survivors. Thus, this effort will inform targeted follow-up recommendations and risk-reducing interventions.

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY/ABSTRACT Cancer remains a leading cause of mortality within the US, responsible for 1 in 4 deaths. Immunotherapies have ushered in a new age of cancer treatment, leveraging the potency of the immune system to restrict cancer without many of the side effects of conventional therapies. These immunotherapies have primarily targeted immune responses specific for tumor antigens (tAg). Although tAg-specific T cell immunotherapies have proven powerful tools in combatting cancer, their success is context-dependent, displaying limited efficacy in most solid tumors. This is largely owed to chronic T cell receptor (TCR) binding to cognate tAg, leading to permanent cellular dysfunction. Solid tumors comprise the majority of cancer cases and deaths, thus, it is essential to exploit other cellular modalities in immunotherapies. Recently, it has become apparent that tAg-nonspecific “bystander” T cells are observed and often outnumber tAg-specific T cells in solid tumors. Although their function within tumors is unknown, bystander T cells can exert cytotoxic effector function once activated by inflammation in a number of contexts. My unique approach leveraging T cells with defined T cell receptors (TCRs) allows me to determine the mechanisms that dictate bystander T cell entry into the tumor and if they maintain the ability to respond to stimulation once tumor-resident. To appreciate the heterogeneity of tumor microenvironments, I will employ multiple animal models of solid tumors. My objectives are two-fold: First, I want to test how bystander T cells migrate to the tumor and if they are spared from dysregulation due to their inability to recognize tAg. Second, I want to test if the dichotomous effects of bystander T cells can be therapeutically leveraged to improve anti- tumor responses. At homeostasis, bystander T cells can simply deny other immune cells access to targets, hindering antigen (Ag)-specific immune responses. Once activated by inflammation, bystander T cells rapidly acquire effector function and directly kill target cells in an innate-like manner. To achieve these objectives, I will employ my expertise in 28-color flow cytometry to interrogate cell phenotype, activation, and functional capacity. I will complement my use of flow cytometry with 3-dimensional immunofluorescence, which will uncover sub- anatomic immune cell organization within the tumor. Hypothesis: My central hypothesis is that bystander T cells in solid tumors remain functional and can be therapeutically leveraged in cancer specifically. I will test this hypothesis through two independent aims: AIM 1: Test the hypothesis that bystander T cells are recruited into tumors by CXCR3 and remain functional in the tumor microenvironment. AIM 2: Test the hypothesis that anti- tumor immune responses can be enhanced by activated bystander T cells or targeted depletion of tissue-resident bystanders. My proposed experiments will elucidate the role of bystander T cells in tumor both with and without interventions, with the goal of developing interventions to improve cancer outcomes, which is in alignment with the mission of the NCI and NIH.

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY / ABSTRACT Cryptococcus neoformans is an opportunistic fungal pathogen that causes localized and disseminated disease, cryptococcosis, in immunocompromised individuals, such as in people living with HIV. A life- threatening manifestation of cryptococcosis is cryptococcal meningitis (CM); a central nervous system infection with acute inflammation of the meninges and the most common cause of meningitis in those with HIV. Sub- Saharan Africa carries the majority of the global CM disease burden and mortality. Because the host immune response is crucial to clearance of the infection and resolution of symptoms, a solid understanding of the host immune response in the state of cryptococcal infection is essential in order to reduce morbidity and mortality. Natural killer (NK) cells are an innate lymphoid immune cell that has been understudied in CM. NK cells have antigen independent cytotoxic ability that render them ideal for targeting both intracellular and extracellular pathogens. This key feature of NK cells could make them an important aspect in the immune response to C. neoformans, which is both an intracellular and extracellular pathogen. Standard antifungal therapy targets the pathogen, yet mortality still occurs, including among persons with sterile CSF cultures. My preliminary data has shown that low CSF concentrations of NK cell cytotoxicity associated soluble molecules are associated with increased risk of acute 14-day mortality in persons with CM. Therefore, improving NK cytotoxic activity may be an innovative therapeutic pathway toward reducing cryptococcal mortality. Additionally, the identification of genes and pathways implicated in impaired NK cell function that could be modulated to improve cytotoxicity could be targets for future therapeutics. My central hypothesis is that NK cells contribute to fungal clearance of C. neoformans, but that a combination of NK cell exhaustion, impaired cytokine production, increased inhibitory receptors, and decreased activating receptor expression due to the underlying HIV infection disrupts their cytotoxic abilities. To test the central hypothesis, I am proposing the following Aims: Aim 1 seeks to compare NK cell natural cytotoxicity and antibody-dependent cell mediated cytotoxicity between HIV-infected persons with CM who die within 14-days of diagnosis and those who survive >14 day. Aim 2 seeks to determine if persons with CM who die within 14-days have high levels of differentially expressed genes involved in exhaustion and inhibition of cytotoxicity pathways when compared to persons who survive CM. Collectively, these findings will provide the first quantification of NK cell cytotoxicity against clinical isolates of C. neoformans and the identification of differentially expressed genes associated with acute CM mortality. This information will provide us with specific genes that are involved in exhaustion, cytokine production, and cytotoxicity that could be modulated in the future with host targeted immunotherapy.

NIH Research Projects · FY 2026 · 2022-02

Project Summary/Abstract The balance and function of effector T cells, regulatory T cells, and anergic T cells are critical for maintaining immune homeostasis. Lack of regulatory T cells (Tregs) leads to autoimmunity mediated by self-antigen specific effector T cells whose targets can include tissues of the eye. Although most Tregs originate in the thymus, our previous studies suggest that in response to retinal self-antigens, Tregs can be generated locally in the retina from conventional CD4+ T cells, a process we refer to as "on-demand" Treg generation. T cell recognition of antigen-MHC-II complexes in the absence of costimulatory signals or in the presence of inhibitory signals can induce anergy, a state of functional unresponsiveness and non-proliferation in T cells. Thus, generation of anergic T cells may be another important mechanism for maintaining retinal immune homeostasis. We have observed a small number of T cells and a population of myeloid cells (microglia), a subset of which can act as conventional dendritic cells within the retina. Thus, we hypothesize there is a local, continual generation of retinal self-antigen specific Tregs and anergic T cells within the retina that contributes to retinal immune homeostasis and that the interaction between T cells, retinal microglia, and possibly non- myeloid retinal cells determines the nature and fate of retinal T cells. This hypothesis will be explored in three aims using the R161H mouse model of spontaneous retinal autoimmunity in conjunction with transgenic mice that allow for the tracking and depletion of dendritic cells, microglia, and Tregs. Aim 1: This aim will define the role that retinal microglia, particularly the dendritic cell subset, plays in antigen presentation that leads to autoimmune pathogenesis in the retina. Aim 2: This aim will define and distinguish the roles that resident retinal microglia versus the non-myeloid retinal cells play in production of anergic and regulatory T cells within the retina. Aim 3: This aim will be translational studies using local therapeutic manipulation of retinal microglia or other antigen presenting cells to limit T cell co-stimulation and promote generation of anergic and regulatory T cells to limit inflammatory injury to the retina.

NIH Research Projects · FY 2026 · 2022-01

Project Summary/Abstract This project will create a massive microdata resource comprising the entire population of the United States in 1950. The 1950 Census is ideal for research on aging: people who were young in 1950 can be linked to myriad sources describing their health and well-being from mid to later adulthood, allowing a prospective view of aging. By linking the 1950 Census to recent health surveys, administrative records, and the national death index, investigators can pursue prospective analyses of the impact of early life conditions—including socioeconomic status, parental education, local environment, and family structure—on later health and mortality. The database will cover the entire population with full geographic detail, providing contextual information on childhood neighborhood characteristics, labor-market conditions, and environmental conditions. The 1950 data will enable transformative research to uncover the effects of early life conditions on health and well-being in later life, including cognitive impairment. The database will make a permanent and substantial addition to the nation’s statistical infrastructure and will have far-reaching implications for research across the social and behavioral sciences. The project involves (1) transcribing 8.3 billion keystrokes of data describing the demographic and economic characteristics of all individuals, families, households, and group quarters present in the U.S. in 1950; (2) evaluating data quality through random blind verification and comparison with published census tabulations; (3) converting approximately ten million different open-ended census responses into numeric classifications compatible with previous and subsequent census data; (4) data cleaning, including editing and imputation of inconsistent and missing data values; (5) developing metadata and documentation, including full descriptions of data processing methods, detailed analysis of comparability issues, and comprehensive machine-processable metadata; and (6) incorporating the database into the Integrated Public Use Microdata Series (IPUMS) data access systems for free dissemination to the scientific community. The proposed work will be carried out by a team of highly-skilled researchers with unparalleled expertise and experience in large-scale data creation, integration, and dissemination. The project is a collaboration with the nation’s largest producer of genealogical data. This public-private partnership allows a highly cost-effective use of scarce resources for shared infrastructure for population and health research.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY/ABSTRACT The blood-retina barrier (BRB) protects the retina by limiting extravasation of solutes and immune cells and by providing active transport mechanisms for required nutrients and hormones. BRB dysfunction is implicated in retinal diseases including diabetic retinopathy, choroidal neovascularization, retinal occlusive diseases, uveitis, macular telangiectasia, familial exudative vitreoretinopathy, retinopathy of prematurity, and Coat’s disease. Although the key role of canonical Norrin and WNT7A/B signaling in inducing and maintaining blood-CNS barriers has been identified, highly potent and therapeutically amenable agonists of these pathways that could promote or restore BRB function are lacking. Signal initiation by Norrin requires the receptor Frizzled4 (FZD4) and two co-receptors, low-density-lipoprotein receptor-related protein 5 (LRP5) and tetraspanin-12 (TSPAN12). Here, we use an entirely novel agonist to activate canonical (i.e., beta-catenin-dependent) signaling in endothelial cells. The agonist is a human antibody modality that functionally mimics Norrin and WNT7A/B. The mechanism of action and its efficacy in distinct types of BRB defects are not known. We will conduct cell-based experiments to test the hypothesis that the agonist activates signaling by inducing complex formation of receptor and co-receptor molecules. To understand which types of BRB defects can be restored by the agonist, we will use multiple mouse models with BRB defects that have distinct characteristics. We will determine the efficacy in restoring the BRB in each model, perform transcriptomic experiments and immuno- histochemical analyses to identify mechanism of BRB restoration, and quantify vascular density. BRB defects, e.g., in macular edema, are treated with anti-VEGF or anti-inflammatory drugs. However, anti-VEGF therapies are not effective in all patients, and other patients may develop resistance. The novel agonist used here has an entirely different pharmacodynamic profile compared to anti-VEGF and anti-inflammatory drugs. Therefore, it is important to evaluate this drug candidate for its therapeutic potential.

NIH Research Projects · FY 2026 · 2021-12

Behavior is organized across multiple spatial and temporal scales, ranging from sub-second motor commands over multi-second movement plans to long term foraging patterns. Currently it is unclear how the brain solves this coordination of multiple intertwined temporal demands. While classical neuroscience experiments typically look at or engage a fixed temporal scale or horizon, ethological studies have long focused on the analysis of naturalistic behavior across freely elicited temporal scales. Here we will use novel developed deep learning pose estimation approaches to study the behavior and associated single unit physiology of foraging behavior in freely moving rhesus macaques. First we will establish that timescales of macaque pose behavior encode cognitive variables such as reward expectation. Second we will establish the link between neural and behavioral timescale coordinating in the decision to action axis of the medial prefrontal wall leading to the anterior cingulate cortex by recording multi-region wireless electrophysiology in freely moving rhesus macaques. Third, using embedding and connectivity analysis we will uncover the mechanisms for inter and intra areal timescale coordination to understand how the brain balances temporal scale demands.

NIH Research Projects · FY 2025 · 2021-12

Project Summary Nerve damage is a common affliction that causes sensory and/or motor deficits. Recovery involves a regenerative process in which damaged axons within a nerve fiber must re-extend to the appropriate target tissues, in a process known as target-specific regeneration. This process often fails in humans, leaving patients with chronic health problems. Improving clinical outcomes requires a better understanding of how target-specific regeneration is regulated. We know that components of the nerve support scaffold can guide axon re-extension along simple paths. However, when axons reach nerve branch points, they require more specific guidance mechanisms to differentiate between multiple paths and select the correct one. We have little understanding of what environmental cues guide these decisions, and how they are appropriately interpreted by regrowing axons. The objective of this proposal is to identify cellular and molecular mechanisms that regulate axon targeting decisions to promote target-specific regeneration. I have established the zebrafish vagus nerve as a model to elucidate mechanisms of target-specific axon regeneration. Regenerating vagus axons select between five nerve branches to robustly re-innervate the correct target tissue, although how they do so is not known. I hypothesize that two non-mutually-exclusive mechanisms regulate target-specific regeneration: 1) chemosensation, in which a regenerating axon can interpret spatially patterned chemical guidance cues in the environment that direct its growth; 2) fasciculation, in which a regenerating axon can recognize undamaged axons that are innervating its intended target and use them as a substrate for directed growth. The three aims of this proposal will comprehensively identify how growing axons interact with their environment at the cell biological and molecular levels during target-specific regeneration. In Aim 1, I will combine a novel single-cell chimera regeneration assay with live imaging and genetic and pharmacological manipulations to establish a conceptual understanding of how in vivo axon-environment interactions guide targeting decisions. In Aim 2, I will combine a novel method to label and isolate live neurons based on their innervation target with in vivo and in vitro techniques to precisely measure how axons of each of the five innervation target groups interact with other axons, and with chemical signals, in the environment. In Aim 3, I will combine innervation target-specific neuron isolation with RNAseq and mutant analysis for unbiased identification of molecules that regulate target selection in each of the five innervation target groups. This study will greatly enhance our fundamental understanding of how axons reinnervate their target tissues during regeneration, and provide an important knowledge base to develop improved treatments for nerve damage.

NIH Research Projects · FY 2026 · 2021-12

This project proposes an integrated suite of microscopy and data analysis advances that would enable quantitative, mechanistic analysis of immune-microenvironment dynamics in poor prognosis solid tumors. While immunotherapies are showing remarkable clinical responses in some advanced cancers, to date, their impact on many solid tumors has been modest. This is due, in part, to solid tumor microenvironments limiting the effectiveness of natural immune responses and immunotherapies. Yet, our understanding of the physical and molecular mechanisms governing T cell infiltration, distribution, and function in native tumor microenvironments remains extremely limited. As such, defining key T cell behaviors as a function of complex tumor microenvironments will identify design criteria that can be used to develop novel cell engineering strategies that optimize T cell-centric therapies for solid tumors. To this end the research theme of the U54 Center for Multiparametric Imaging of Tumor Immune Microenvironments (C-MITIE) is to define physical and molecular barriers to effective anti-tumor immunity and immunotherapies through advancement and development of state-of-the-art live cell and tissue optical imaging platforms and quantitative analyses. To achieve our goals, our framework brings together advanced optical imaging platforms, nano- and micro- fabrication, genome engineering, cancer immunology, and biophysical modeling. Thus, from this integrated effort we seek to define mechanisms of immune suppression that guide the development of next-generation cell-based immunotherapies.

NIH Research Projects · FY 2025 · 2021-11

Project Summary Cryptococcal meningitis (CM), caused by the opportunistic fungal pathogen Cryptococcus neoformans (Cn), is a leading cause of HIV-related mortality worldwide. In healthy individuals, exposure to Cn in early childhood results in a pulmonary latent infection that is asymptomatic, but leads to the formation of lung granulomas. Following HIV-associated compromise of the immune system, control of latent Cn infection within pulmonary granulomas is lost and the fungal pathogen disseminates to cause meningitis. The host immune cells and effector functions critical for establishing and maintaining control of latent Cn infections have not been identified. Clearance of latent infections is warranted in the context of advanced HIV care. Understanding how the healthy immune response controls latent Cn infection is needed to: 1) define critical immune functions that prevent disease, 2) determine why a healthy immune system is unable to eradicate latent infections, and 3) develop targeted therapies that mitigate disease progression in HIV patients with CM. Using a novel mouse inhalation model of latent cryptococcosis developed in our lab, I will test my central hypothesis that a Th1 CD4 T-cell response is necessary and sufficient to control the latent pulmonary Cn infection, but is unable to clear the infection due to intrinsic deficiencies in interferon-γ (IFNγ) signaling caused by persistent Cn survival within granulomas. In Aim 1, I will combine laser capture microdissection with RNA sequencing to determine the cellular and effector functions responsible for containing Cn within granulomas. In Aim 2, I will use a mouse model that mimics HIV-induced CD4 depletion and post-mortem granulomatous lung tissue biopsies from HIV patients with CM to investigate how HIV-induced loss of CD4 T-cells disrupts control of latent Cn infection within granulomas. In Aim 3, I will use adoptive T-cell transfers and exogenous IFNγ supplementation to elucidate the CD4 T-cell subset and effector functions responsible for controlling latent Cn infection. The long-term goal of these studies is the development of immune-regulated therapeutic strategies for HIV- associated CM. These strategies include clearing latent Cn infection prior to HIV-immunosuppression in at-risk individuals and mitigating disease progression in HIV/AIDS patients by replacing the essential Cn-specific CD4 T-cell subset required for control of Cn infections. This proposal will lay the necessary groundwork for developing therapies that specifically target Cn infection, but avoids eliciting immune reconstitution inflammatory syndrome in HIV/AIDS patients with CM. The proposed research and training plans provide a rigorous program for successful completion of MD- PhD degrees, and will further my development into a successful academic infectious disease physician scientist who drives cutting-edge translational research in HIV/AIDS and HIV-associated opportunistic pathogens.

NIH Research Projects · FY 2025 · 2021-09

Project Summary 2024: Benchmarking Project The SenNet consortium is working together to develop and apply multiple spatial imaging technologies to investigate the molecular and cellular heterogeneity of cellular senescence in aged tissues. However, one major obstacle faced by the consortium is the uncertainty surrounding the ability of the various technologies being employed to robustly characterize senescent cells in different tissues. To address this challenge, the SenNet consortium will undertake a collaborative effort to test the effectiveness of various spatial omics technologies on the same human tissue across different sites, as well as to test their effectiveness across different tissues. This endeavor aims to accomplish several goals: firstly, to determine the robustness of senescence-associated marker detection based on the platform, secondly, to extract diverse information about senescence from the same tissue. This consortium-wide benchmarking project will focus on spatial omics techniques that offer single- cell resolution. This approach is based on the current understanding that senescent cells are present in relatively low abundance in aged tissues and the need to use technologies that can accurately detect individual senescent cells and differentiate them from surrounding cells. By rigorously testing the robustness of high-plex technologies already in use within the SenNet Consortium across multiple sites and tissues, we will determine which techniques are most accurate and informative in detecting senescent cells in different tissues.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Current treatments for addiction remain limited by a gap in the fundamental knowledge of how different regions of prefrontal cortex interact in decision-making. Here, we examine the direct neural correlates in prefrontal cortex of the control and costs of effortful decision making in humans at high spatiotemporal resolution, laying the groundwork for the development of new treatments to improve decision making in addiction. This K23 Career Development Award aims to provide me with the necessary training to become an independent investigator translating intracranial studies of decision making into non-invasive biomarkers and targets for neuromodulation in addiction. Toward this end, I propose the following training objectives: (1) Develop advanced skills in neural decoding from intracranial electrophysiology data; (2) gain expertise in designing neuroeconomic and computational psychiatry experiments; and (3) gain expertise in utilizing brain stimulation in human behavioral experiments. The overall research objective of the proposed project is to resolve the roles played by prefrontal regions during effortful decision making by combining a neural decoding with functional connectivity analysis and with cortical stimulation to perturb network function. The central hypothesis is that anterior cingulate cortex (ACC) allocates cognitive resources for control, based on the cost of control assessed in orbitofrontal cortex (OFC), in turn determined by the efficiency evidence accumulation in dorsolateral prefrontal cortex (DLPFC). The specific aims of this research are to (1) Dissociate the roles of three prefrontal regions in the regulation of cognitive control. (2) Map the prefrontal network allocating cognitive effort, and (3) Causally dissect the cognitive effort network. Innovation: (1) Method: Use of neural decoding, information theory with intracranial electrophysiology and stimulation to characterize prefrontal networks in humans; (2) Design: Integration of experimental paradigms from neuroeconomics with computational models of neural processing and neurophysiology; (3) Concept: Measuring and modulating cognitive effort through simultaneous recording and stimulation of prefrontal regions during effortful decision-making. The proposed research is significant because it resolves a controversy over the functional roles of key regions of prefrontal cortex in effortful decision making that are putative targets for neuromodulation in addiction. The new fundamental knowledge generated by this proposal will lay the foundation for the development of novel biomarkers and targets for improving cognitive control and decision making in addiction with mechanism-based computationally guided neuromodulation.

NIH Research Projects · FY 2025 · 2021-09

Neurodevelopmental processes are shaped by dynamic interactions between genes and environments. Maladaptive experiences early in life can alter developmental trajectories, leading to harmful and enduring developmental sequelae. Pre- and postnatal hazards include maternal substance exposure, toxicant exposures in pregnancy and early life, maternal health conditions, parental psychopathology, maltreatment, and excessive stress. To elucidate how various environmental hazards impact child development, it is imperative that a normative template of developmental trajectories over the first 10 years of life be established based on a sufficiently large and demographically heterogeneous sample of the US population. To accomplish this, the Healthy Brain and Child Development (HBCD) Consortium has been formed to deploy a harmonized, optimized, and innovative set of neuroimaging (MRI, EEG) measures complemented by an extensive battery of behavioral, physiological, and psychological tools, and biospecimens to understand neurodevelopmental trajectories in a sample of 7,200 mothers and infants enrolled at 27 sites across the United States (US). The HBCD Study will carry out a common research protocol under direction of the HBCD Consortium Administrative Core (HCAC) and will assemble and distribute a comprehensive and well-curated research dataset to the scientific community at large under the direction of the HBCD Data Coordinating Center (HDCC). The overarching goal of the HBCD Study is to create a comprehensive, harmonized, and high-dimensional dataset that will characterize typical neurodevelopmental trajectories in US children and that will assess how biological and environmental exposures affect those trajectories. A special emphasis will be placed on understanding the impact of pre- and postnatal exposure to opioids, marijuana, alcohol, tobacco and/or other substances. To address these broad objectives, the sample of women enrolled will include: 1) a varied cohort that is representative of the US population; 2) pregnant woman with use of targeted substances (opioids, marijuana, alcohol, tobacco); and 3) demographically and behaviorally similar women without substance use in pregnancy to enable valid causal inferences. In addition, the HBCD Study will identify key developmental windows during which both harmful and protective environments have the most influence on later neurodevelopmental outcomes. The large, multi-modal, longitudinal, and generalizable dataset that will be produced for the first time by this study will provide novel insights into child development using state-of-the-art methods. The HBCD Study will inform public policy to improve the health and development of children across the nation. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions to the national opioid public health crisis. The NIH HEAL Initiative bolsters research across NIH to improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Mucosal surfaces account for the vast majority of transmission events for human retroviruses – e.g., human immunodeficiency virus type 1 (HIV-1) and human T-cell leukemia virus type 1 (HTLV-1). HIV-1 and HTLV-1 infection through the oral cavity represents a significant gateway for postnatal transmission from mother to child. Virus spread via cell-to-cell transmission aids in establishment of viral infection in a newly infected individual. While significant advancements in our understanding of retrovirus replication have been made, there are many details that remain poorly understood. For instance, it is still unclear how virus particle assembly sites are determined and how viral structural proteins and genetic material translocate to these locations on the inner leaflet of the plasma membrane. The details of viral particle assembly are particularly unknown in the context of cell-to-cell transmission. In this application, I propose to use molecular virology and state-of-the-art imaging approaches to elucidate mechanisms of cell-to-cell transmission of different human retroviruses by comparative analyses. It is well documented that HTLV-1 is efficiently transmitted via cell-cell contacts, i.e., the virological synapse (VS). This is likely also the case for HIV-1 but has been commonly underappreciated. Virus transmission at cell-cell contacts via the formation of a VS can result in polarized virus particle release into the VS. Virus particle formation is driven by the Gag structural protein, which multimerizes at the virus assembly site (e.g., points of cell contact), resulting in particle biogenesis and release. In preliminary studies, our lab has observed that the pool of Gag utilized in particle biogenesis in non-polarized cells was primarily recruited from the plasma membrane for HTLV-1 whereas for HIV-1 Gag is recruited from the cytoplasm. This fundamental observation of differential modes of Gag recruitment to particle assembly sites may help explain the distinct reliance of cell-to- cell transmission as a productive mode of virus spread for HTLV-1 compared with that of HIV-1. I propose 2 lines of investigation for this application. I will first investigate whether the differences in HIV-1 and HTLV-1 Gag puncta biogenesis observed in non-polarized cells are also conserved in polarized cells. Second, I will investigate virus- host cell interactions that help facilitate human retrovirus assembly, particularly in the context of cell-cell contacts, which is of particular significance in oral biology. An important aspect of this aim will be the use of novel and innovative technology, cryogenic-correlative light and electron microscopy (cryo-CLEM), in order to gain greater insights into the role(s) of host cellular proteins important for virus assembly. Human retrovirus particle assembly is a critical step in infectious virus transmission, including oral transmission at mucosal surfaces in the context of cell-cell contacts. These studies will help contribute new information to better understand key aspects of human retroviral replication that are not fully understood and represent knowledge gaps in the field of virology and will provide critical information to aid in the prevention of retroviral transmission through the oral cavity.

NIH Research Projects · FY 2025 · 2021-09

Abstract The 10,000 Families Study (10KFS) will be a new, family-based cohort study in Minnesota, the ‘Land of 10,000 Lakes’. Due to our high incidence rates of hematologic malignancy (leukemia, lymphoma, and myelodysplastic syndromes), we will focus our proposal on environmental exposures of concern that are possible hematologic carcinogens with limited evidence in humans as defined by the International Agency for Research on Cancer (IARC). This includes glyphosate, poly- and perfluoroalkyl substances (PFAS), and radon, all of which have well-described geographic variation in prevalence in the state of Minnesota. During the UG3 phase, we will evaluate innovative exposure assessment methodologies and recruit diverse participants into the 10KFS cohort. Exposure assessment will include residential history, ambient measures in air and water at current residence, as well as individual-level exposure assessed with silicone wearable bracelets, serum, hair, and urine. Participants will be recruited from targeted counties that maximize the exposure distribution to our three carcinogens of interest and include under-represented populations from rural and immigrant communities. We will measure clonal hematopoiesis of indeterminate potential (CHIP), a precursor to heme malignancy, and perturbations in immunity and epigenetics as our primary cancer-related outcomes in the UH3 phase. Our target is to recruit 8,750 participants from 4,000 households to participate using recruitment strategies that have been developed and pilot tested. Our central hypothesis is that PFAS, glyphosate and radon contribute to increased hematologic malignancy specifically, and cancer incidence broadly, in Minnesota. Cancer incidence will be determined via annual record linkage with the Minnesota Cancer Reporting System (MCRS) and the Virtual Pooled Registry Cancer Linkage System. The scientific aims in the UG3 phase will be to: 1) evaluate innovative exposure assessment methods for PFAS and glyphosate that can be easily implemented in a large scale study and 2) describe the exposure burden in under-represented immigrant populations. We will complete recruitment of the study population in the UH3 phase where the specific aims are to: 1) determine whether levels of radon, glyphosate, and PFAS are associated with increased prevalence of CHIP and 2) utilize epigenomics to test the hypotheses that radon, glyphosate and PFAS exposure are associated with immune dysregulation and epigenetic signatures. Our community engagement will be conducted in partnership with the non-profit research institute Hispanic Advocacy and Community Empowerment Through Research (HACER), the Minnesota Freemasons and Order of the Eastern Star and community educators from the Extension Center for Family Development. The results of this study will provide needed data to inform whether these agents are hematologic carcinogens, it will address documented disparities in exposure that occur within our state and provide a mechanism for community outreach and education concerning these exposures and any potential cancer link.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT – Overall A growing body of research documents the profound contribution of structural and interpersonal racism to disparities in chronic disease including cardiovascular disease (CVD) and the related chronic conditions (RCC) of hypertension and obesity. In Minnesota, where the murder of Mr. George Floyd at the hands of police instigated a local, national, and global reckoning on the pervasiveness of racism, there is an urgent need to understand and ultimately address the ways that racism undermines well-being of Black, Indigenous, and people of color (BIPOC) communities. The mission of our proposed Center for Chronic Disease Reduction and Equity Promotion Across Minnesota (C2DREAM) is to reduce disparities in CVD and RCC experienced by BIPOC communities including immigrants and refugees across Minnesota. Our 3 proposed intervention projects target community and primary care approaches to diet, physical activity, smoking cessation, and other proximal contributors to CVD and RCC among Minnesota’s diverse BIPOC communities. C2DREAM will promote innovative, multilevel interventions that target fundamental causes of health inequities among diverse BIPOC communities across Minnesota; generate knowledge, analytical and implementation approaches, and community engagement strategies to understand and develop solutions to address racism at multiple levels; develop and support the next generation of health equity researchers; create and sustain a community-engaged and equity-guided dissemination and implementation processes to translate research results and knowledge regarding addressing chronic disease inequities into action; and employ equity-guided evaluation approaches. All center activities and research projects are aligned with a unifying conceptual model guided by the NIMHD’s Minority Health & Health Disparities Research Framework. C2DREAM is a regional partnership that spans the state among the University of Minnesota (UMN), including the Medical School on the Twin Cities (central) and Duluth (northern) Campuses, and the School of Public Health; Mayo Clinic including Rochester and community-based Mayo Clinic Health System clinics (southern); and Hennepin Healthcare, a county-owned HMO and the state’s largest safety net hospital (central). Our partnership builds on many formal and informal connections across collaborators and institutions, including NCATS-funded UMN Clinical Translational Science Institute (CTSI) and Mayo Clinic Center for Clinical and Translational Sciences (CCaTS) resources, as well as extensive partnerships with key community stakeholders. Our team has rich expertise in measurement of racism at multiple levels, health promotion interventions targeting CVD and related chronic conditions, behavioral medicine, epidemiology, public health, psychology, community based participatory research, policy, communications, and implementation science. This breadth, depth, and regional reach uniquely positions us to rigorously investigate fundamental drivers of health inequities and generate solutions that will impact Minnesota and the nation.

NIH Research Projects · FY 2025 · 2021-09

Despite the prevalence of prostate cancer, the current tools available to manage the disease continues to leave physicians and their patients in a position to overdiagnose and overtreat. The confidence to pursue more conservative approaches like active surveillance are limited, as biopsy is known to underestimate the grade and extent of disease, both of which are important for risk stratification. Targeted biopsies, by means of MRI- guidance, are becoming the preferred way to ensure the most aggressive appearing lesions are sampled in the hopes of avoiding some of the issues with standard biopsy approaches. These targeted biopsies make use of multi-parametric MRI (mpMRI) which includes both anatomical and functional information that are complimentary and together increase the sensitivity and specificity for cancer detection. However, the ability to effectively use mpMRI requires specialized training while the standards for properly using the multiple MRI datasets are still being developed. To address this issue, we have developed an alternative method that would provide a quantitative, user-independent, summary of the mpMRI data (qMRI) to visually “map” disease and assess its aggressiveness. Using quantitative MRI, a Composite Biomarker Score (CBS) map is generated, with a demonstrated increase in sensitivity and specificity for tumor detection compared to any single qMRI parameter. Our primary goal is to integrate this predictive qMRI model into a computer-aided diagnostic (CAD) system (referred to as CBS-CAD) to improve the use of mpMRI in PCa management. Employing quantitative MRI (qMRI) can address the issues of a qualitative image analysis if the major roadblocks to its adoption can be overcome. To address the roadblocks and implement the CBS-CAD system we will pursue the following specific aims: 1) develop an analysis pipeline to evaluate qMRI performance and translate CBS-CAD methods, 2) perform a multi-vendor, multi-site quantitative imaging technical performance evaluation and 3) perform a multi-center clinical validation study assessing CBS-CAD performance. Our expected outcome of this academic-industry partnership will be the integration of several novel technologies into a comprehensive CAD system consisting of a phantom and automated software for 1) qMRI system validation and 2) clinical translation of novel models for detecting cancer and assessing aggressiveness.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Many diseases are complex, heterogeneous, conditions that affect multiple organs in the body and depend on the interplay between several factors that include genetic, cellular, molecular, and environmental factors. It is therefore not surprising that the pathogenesis of many complex diseases remain elusive, and therapeutic targets are lacking. The traditional approach that focus on a small number of molecules (e.g., genes or metabolites) or a single type of data (e.g., clinical or genetic) cannot address this complexity and heterogeneity. Integrative or systems biology approaches and network analysis can be used to leverage the strengths of data from multiple sources (e.g., genomics, metabolomics, epidemiology, clinical data) to achieve new insights into the pathobiology of complex diseases. Recent technological advances have enabled the production of vast amounts of diverse but related data with rich information that offer remarkable opportunities to understand biological processes involved in complex diseases and to transform medicine, yet at the same time present significant analytical challenges including how to effectively synthesize information from the tens of thousands of data points to identify important biomarkers with potential to serve as therapeutic targets. To alleviate this, we will develop and apply a suite of novel, robust, and powerful statistical and machine learning methods for the integration and interpretation of cross-sectional and longitudinal data from multiple sources. These models will also be used to define subpopulations of patients who have different prognoses or require different therapeutic approaches based on data from different sources. Further, we will make use of recent advances in network theory to model the complex multilateral relationships in molecular data from multiple sources. The proposed methods will be applied to several publicly available datasets and cohorts to ensure that we can generalize our work to other datasets and cohorts and thus increase the long-term impact of our research. The proposed research will also contribute valuable statistical and machine learning algorithms that will be broadly applicable to data from multiple sources and multiple cohorts and will be made available to the public free of charge.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Chemical toxicants and carcinogens present in the polluted air are the likely causative agents in the associated cancers of aerodigestive tract in exposed individuals. Understanding the relationship between the level of exposure to these pollutants and the risk of cancer is key to identifying individuals or populations at risk and informing the development of preventive measures. The overarching goal of this study is to advance such understanding through a collaborative research between the Masonic Cancer Center, University of Minnesota and Centre for Cancer Epidemiology (CCE) in Mumbai, India. We will employ a panel of established and novel air pollution-related biomarkers to test our overall hypothesis that the uptake of air pollution-related chemical carcinogens is associated with the risk for LC and HNC in Indian nonsmokers. In Aims 1 and 2, we will conduct case-control studies to assess the association of air pollution-related biomarkers in plasma with LC and HNC, respectively, in Indian nonsmokers. To achieve the goals of these Aims, we will use available biological samples and questionnaire data from the corresponding epidemiological cohorts conducted by the CCE. We will develop capacity for biomarker measurements at CCE by transferring the U.S. team's expertise in this area. In Aim 3, we will recruit healthy nonsmokers with various levels of usual exposures to air pollution, based on their occupation or lifestyle. We will compare a panel of biomarkers of exposure and effect across the groups and correlate to pollutant levels collected through personal air sampling devices worn by a subset of participants. We will use specially designed air samplers to characterize the chemical profile of various air pollution scenarios. This aim will aid in the interpretation of biomarker data generated in Aims 1 and 2, and will inform future biomarker-based studies of air pollution in India. An important outcome of this study is the development of capacity for future biomarker research of cancer risk in India using CCE cohorts. Such research can be further expanded to other population subgroups (e.g., smokers or occupationally exposed individuals) and additional environmental and dietary exposures in India.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Diabetes has a tremendous impact on the health and well-being of affected individuals, as well as a considerable overall societal burden. Pancreatic islet transplantation has the potential to cure diabetes, but one of the main problems limiting the success of this treatment is an inadequate supply of islets. Islets from a single donor are often insufficient to achieve insulin independence, and multiple infusions are often required, each with increasing risk. Two potential strategies exist to increase the number of islets available: (1) pool islets from multiple donors and perform single procedure, high-dose transplants; and (2) develop alternative sources such as stem-cell-derived islets. The availability of these limited resources becomes a supply chain problem, and for either approach, a method for islet preservation is essential. Our long-term objective is to develop a method for cryopreserving, or “banking,” islets prior to transplant. No previous strategy has achieved the high viability, function, and clinical scalability required for transplant in a single approach. To achieve long-term islet banking, we propose to use an alternative cryopreservation strategy, vitrification. That is, cryogenic storage in an ice-free glassy state. A significant challenge in the vitrification of biospecimens is that the cooling and heating rates needed for vitrifying and rewarming are tremendously high (>107 °C/min). These rates are reduced by adding cryoprotective agents (CPA) that inhibit ice formation, but these agents are themselves toxic to islets. Thus, the critical challenge in islet vitrification is achieving fast enough cooling and warming to avoid ice, while avoiding toxicity from the CPA, and doing so in a clinically scalable manner. Using engineering principles of heat and mass transfer, our multidisciplinary research team has developed an approach for vitrification and rewarming (VR) to solve this problem, termed “cryomesh VR,” for islets. Our central hypothesis is that the improved heat transfer achieved by cryomesh VR, combined with optimizations in CPA use, will enable ice-free vitrification and rewarming of islets while avoiding toxicity. Our preliminary data achieve cooling and warming rates far exceeding other methods, and we have shown CPA loading and unloading protocols with low toxicity in mouse, human, pig, and human stem-cell-derived (SC) islets. Indeed, in all cryopreserved islet models tested we have achieved viability, recovery, and function that meets or exceeds all previous reports and does so in a clinically scalable method. To further improve our approach and move towards clinical translation, we propose the following aims: Aim 1. Refine the optimal physical conditions for the cryomesh VR of mouse, human, and SC islets; Aim 2. Measure the viability, function, and in vivo potency of mouse, human, and SC islets following cryomesh VR; Aim 3. Define the molecular and cellular changes occurring in response to cryopreservation; and Aim 4. Scale-up cryomesh VR for clinical throughput and adapt the processes for cGMP production. If successful this approach could revolutionize how islets are isolated, allocated, and stored prior to transplant and increase utilization of deceased donor pancreases for the cure of diabetes.

NIH Research Projects · FY 2025 · 2021-09

Squamous cell carcinoma of the head and neck (HNSCC) is a devastating, frequently disfiguring, and often fatal disease, expected to affect more than 53,000 people in the U.S. in 2020 and kill more than 10,000. Prevention of this terrible disease is critical. Cigarette smoking, alcohol consumption, and human papilloma virus (HPV) are well established major causes of HNSCC; only smoking and alcohol consumption are considered here. Cigarette smoking and alcoholic drinks are sources of multiple DNA adducts that are critical in the carcinogenic process. This proposal will establish a liquid chromatography-nanoelectrospray ionization- high resolution tandem mass spectrometry (LC-NSI-HRMS/MS) profile analysis of 12 oral cell DNA adducts that are likely causes of HNSCC. This was inspired by our recent analysis of DNA adducts in oral cells, in which we found levels more than 20 times higher in cigarette smokers than in non-smokers. These exciting results encouraged us to propose a profile analysis of important carcinogen-derived DNA adducts in oral cells according to the following specific aims: 1. Develop an LC-NSI-HRMS/MS profile analysis method for quantitation of 12 important and representative carcinogen and toxicant – DNA adducts in human oral cells and tissue. The adducts are derived from various carcinogens and DNA reactive compounds in cigarette smoke and alcoholic beverages. 2. Apply the profile analysis to oral cells from currently healthy individuals: a) 100 non-smokers who are non- drinkers or light drinkers; b) 100 cigarette smokers who are non-drinkers or light drinkers; and c) 100 cigarette smokers who are moderate or heavy drinkers. Comparisons of adduct levels in groups a and b will identify adducts enhanced by cigarette smoking while comparisons of groups b and c will identify adducts that are enhanced by the combination of smoking and moderate or heavy drinking. 3. Test the longitudinal stability of the oral cell DNA adduct profile analysis over a 6 month period in 50 smokers who are non-drinkers or light drinkers. 4. A) Determine the DNA adduct profile in oral cells collected from 75 smokers with HNSCC and compare to that in 200 smokers without HNSCC recruited in Specific Aim 2 with the goal of identifying an adduct profile that is characteristic of HNSCC incidence. B) Compare the oral cell DNA adduct profile from part A of this aim to that in tissue, both normal and tumor, in a subset of 60 patients from part A who undergo surgery, to determine whether oral cell DNA adduct patterns are consistent with those in tissue. Our results will potentially identify individuals who are susceptible to HNSCC but are unable to quit smoking. Once identified, aggressive lifestyle and monitoring interventions in these subjects such as oral examinations 2-4 times per year can be initiated for prevention or early detection of this disfiguring and often fatal cancer.