University Of Wisconsin-Madison

NSF Awards · FY 2024 · 2024-10

An organism’s ability to respond to its environment is fundamental to its survival. When the organism of interest is involved in a host-pathogen interaction, “the environment” becomes complex, including includes not only external forces, but also the conditions imposed by the interacting partner. There are also two genomes at play, each of which affects the behavior of both organisms. In addition, external environmental forces impose additional pressures, and each genome can affect how both partners respond. This project examines the role the host genome plays in altering behavior of both host and pathogen under environmental stress. It leverages an agriculturally relevant system, nematode infection of tomato. Parasitic nematodes are responsible for around $125 billion in annual crop loss worldwide with yield loss upwards of 80% for tomato. Limited control options are available, and the situation is exacerbated by an emerging concern in agriculture: the effect of warming nighttime temperatures (WNT). This unprecedented trend is causing critical challenges to crops. Broader, future impacts of this work include the development of novel approaches to examine host-pathogen interactions and how they are affected by external conditions. This then will lead to the identification of plant lines that are more resilient to both abiotic and biotic stresses. Importantly, by elucidating the molecular biology behind the parasite response to those plants under WNT, this study will go beyond merely identifying relevant host genes to contribute new insight into the mechanisms by which those genes alter the nematode biology. Understanding the nematode in addition to the plant paves the way towards targeting the parasite directly for crop improvement. The goals of this project align with an overarching concern in genetics: to identify DNA variants that influence how individuals respond to their environment. Here, the concept of “individuals” and “environment” are complex. DNA variants in one species will be identified that, in tandem with external environmental conditions, affect how another, interacting species responds. The environmental context considered is warming nighttime temperatures (WNT), a critical, highly relevant, and current environmental concern. Genetically variable tomato plants derived from a cross between a cultivated line and a wild line will be infected with a genetically homogeneous strain of parasitic nematodes. A control experiment will also be performed with uninfected plants. These early, late, and control experiments will be carried out under two temperature regimes: normal nighttime temperatures and WNT. For each treatment combination, phenotypes related to infection and plant health will be collected, along with gene expression data for both plant and nematode. With this design, connections between DNA variants in the tomato genome and molecular responses of the nematode as well as the plant will be made, and the effect of WNT on these connections will be uncovered through via a series of genetic mapping experiments. Leveraging the connections identified in this way, more complex genotype-expression-phenotype pathways can subsequently be inferred, providing a detailed view of the molecular biology of the plant-parasite interaction response to WNT. It will also pinpoint promising candidate genes, which will be functionally validated. All project outcomes will be made publicly accessible through publications and deposition of data and resources in long-term repositories. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-10

By collecting and storing genes from important wildlife now while these wildlife populations are under current threat of diminishment and possible extinction, current conservation practice can support efforts of future generations to restore these populations and/or explore them for potential benefits for the ecosystem. This project explores the proactive, large-scale storage of the genes of wildlife of cultural, conservation, and scientific significance. The approach seeks a unified mode for collecting, storing, and recording collections, based on a consultative process that honors the intentions, preferences, and priorities of local communities, including and especially Indigenous populations. The aims of the project include: 1) exploration of minimally-invasive sampling and storage of genetic samples from American red fox and American pine marten in the Great Lakes region; 2) evaluation of the generation of induced pluripotent cells (iPSCs) from these samples, a key technique to support species recovery; 3) evaluation of storage techniques (pooled-sample approaches) to reduce space and energy consumption; 4) execution of consultative engagements with communities, including Indigenous partners to evaluate the role of such technologies, if any, in local conservation planning, and to develop a unified system for recording and communicating collections, and 5) provision of training and instrumentation for communities to collect, process and store data for their own purposes, to assure self-determination. The project will conduct preliminary fieldwork to collect 15 minimally invasive genetic samples from red foxes (Vulpes vulpes) and American martens (Martes americana) in the Great Lakes Region. The samples will be used to identify technical gaps in generating induced pluripotent cells (iPSC) (i.e., reprogrammable cells that preserve genetic diversity) and to assess the coverage of genetic variation in the biosamples relative to the wild population. This proof-of-concept study will also identify technical gaps in long-term cryogenic preservation of biosamples and procedures to re-isolate single iPSC clones representing sampled individuals. We will further develop and deploy an ethical framework that honors Traditional Knowledge, Indigenous Data Sovereignty (IDS) principles, and public trust. The project will host a series of workshops, convening diverse publics from the Great Lakes region, including Sovereign Native Nations, to discuss and develop: 1) Data sovereignty principles as they apply to genetic material, 2) Governance guidelines for data sharing, storage and ownership of genetic information; 3) Training and infrastructure needs of communities and sovereigns related to genetic data management, 4) Biosampling decision making. Workshop results will include action plans for policy, training and educational needs, and guidelines for collaboration. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY/ABSTRACT Hospitalized children experience alarming rates of harm due to medical errors and rely on their clinicians and parents to exchange important information about their care and work together to identify and intercept errors. While recent mandates require clinicians to share their clinical notes with parents, which detail a child’s diagnosis, progress and treatment plans, fewer than 10% of parents read these notes during their child’s hospital stay. To address this gap, we developed Bedside Notes – a health IT intervention that provides parents with real-time access to their child’s clinical notes through an inpatient portal on a bedside tablet during their child’s hospitalization. Our single-center pilot study showed a 14-fold increase in parent access to notes after implementation. Additionally, one in five parents who accessed notes identified a potential safety concern, 60% of which were confirmed. The long-term goal of this research is to improve access to trusted health information for parents of hospitalized children to enhance the safety of pediatric inpatient care. The objective of this proposed study is to evaluate the impact of the Bedside Notes intervention on note access and parent- reported outcomes for hospitalized children. Our objective will be accomplished with these aims: 1) Assess the impact of the Bedside Notes intervention on parent access to inpatient notes, 2) Evaluate the effectiveness of note access on improving parent safety perceptions and experiences, and 3) Identify contextual factors contributing to parent use of and hospital staff receptivity to the intervention. Guided by the Systems Engineering Initiative for Patient Safety 2.0 framework, our approach will be to conduct a hybrid type 1 randomized controlled trial (RCT) with English and Spanish-speaking parents of 558 children admitted to a pediatric service at 3 sites. Parents will be randomized to one of two arms: (1) usual care (notes through an outpatient portal on a personal device) or (2) the Bedside Notes intervention (notes through an inpatient portal on a hospital-owned bedside tablet + orientation video). Mixed methods (EHR audit reports, surveys, interviews) will assess note access and parent safety perceptions and experiences. Deliverables include data on the efficacy of the Bedside Notes intervention and contextual factors informing future implementation and dissemination. Our team’s expertise in pediatric health services research and consumer-facing informatics, and experience successfully collaborating to design and pilot test the Bedside Notes intervention, form a strong foundation for this research. This proposal is innovative because it will be the first RCT evaluating an intervention to improve note access and impact on parent-reported outcomes. Utilizing a multisite RCT design, multiple electronic health record vendors, and national advisory group feedback, this study is poised to make a sustained positive impact on the delivery of patient- and family-centered care. This study is significant because findings will advance scientific knowledge regarding the potential to leverage health IT to engage parents as partners in improving the safety of inpatient care for children, an AHRQ priority population.

NIH Research Projects · FY 2025 · 2024-09

Uncorrectable vision loss, a.k.a. low vision, can significantly impact activities of daily living (ADLs), affecting indepe ndence, health, and social well-being . Magnifiers are the most commonly used low vision aids, however, they largely reduce visual field. Augmented reality (AR) glasses present a unique opportunity f or low vision technology by intelligently recognizing the environment and enhancing users' vision based on their contexts. However, existing AR solut ions mainly rely on image processing techniques, generating pure pixel-level augmentations to the whole scene, such as pixel remapping for visual field loss, and recoloring based on distance. These solutions arbitrarily alter the user's full visual field with no semantic understanding of the scene, distorting users' natural vision and diminishing important visual details. As a result, they ca nnot effectively support complex ADLs that involve constant motion and object interactions, such as cooking in the kitchen, or safely navigating a crowded street. The overall objective of this proposal is to fundamentally advance low vision technology by leveraging state-of-the-art Al techniques and presenting unobtrusive scene-aware AR augmentations that are tailored to users' environments and visual abilities. Contextualized in two key ADLs (meal preparation, outdoor mobility), the research will investigate how to recognize and augment visual elements in AR from three fundamental dimensions via three specific aims: (A1) E nhancing perception of visual salience, (A2) E nhancing object affordance for safe and efficient interactions, (A3) E nhancing awareness of environmental dynamics. In each aim, the research will contribute to both HCI and Al by designing scene-aware AR augmentations with low vision users and rehabilitation professionals, and developing egocentric video datasets and Al models to enable fast and robust scene recognition in AR. In the final year, a holistic AR system will be developed and evaluated for real world impact and limitations via field studies (A4). With the Al-powered AR systems f or low vision, the project will contribute to the mission of N EI by fundamentally transforming conventional low vision aids and improving independence and quality of life for low vision people in various challenging ADLs. The involvement of low vision participants and rehabilitation professionals throughout the research will also increase their exposure to state-of-the-art technolog y, potentially increasing the technology acceptance and adoption in low vision rehabilitation. RELEVANCE (See instructions): The proposed research is relevant to public health because it investigates how to design and develop Al-powered AR systems to enhance low vision users' visual perceptions, empowering them to navigate activities of daily living with greater safety, efficiency, and independence. The project's contribution to low vision technology directly aligns with the mission of N EI , which emphasizes the importance of eliminating vision loss and improving quality of life through vision research.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT Heart disease is the leading cause of mortality globally; yet prevailing treatment modalities fail to address the fundamental issue of replacing permanently lost contractile myocardium. Human pluripotent stem cells (hPSC) have the ability to generate large numbers of human cardiomyocytes (CMs) for cell therapy, drug toxicity, and disease modeling. However, current hPSC-CM differentiation protocols suffer from significant batch-to-batch and line-to-line variability in reproducibly generating pure hPSC-CM populations. One explanation for this variability is that existing hPSC-CM differentiation protocols rely heavily on murine cardiogenesis biology, from which humans evolutionarily diverged 100 million years ago. Ethical concerns and the paucity of embryonic tissue preclude studies of in vivo human cardiogenesis. However, hPSC-CM differentiation provides the intrinsic opportunity to define the molecular events that govern efficient human cardiogenesis by temporally comparing high and low purity hPSC-CM differentiations. We have demonstrated that high and low purity hPSC-CM differentiations have distinct signal transduction, metabolic fuel consumption, and transcriptional regulation as early as the mesoderm progenitor stage using bulk transcriptomics, proteomics, and metabolomics. In this study, we propose to use single nucleus RNA and ATAC sequencing to objectively define the heterogeneous cell populations present during progenitor stages of high and low purity hPSC-CM differentiations in multiple genetically distinct cell lines. By comparing batch-to-batch and line-to-line variability in hPSC-CM differentiation purity, we expect to reveal conserved signaling pathways and gene regulatory networks that dictate human CM and non-CM cell specification. After uncovering the molecular mechanisms underlying divergent developmental cascades, we will apply this knowledge to enhance the robust generation of high purity hPSC-CMs. To accomplish this feat, we will use stage-specific signaling approaches to improve the production of high potency mesoderm and cardiac progenitors, which are poised to become high purity hPSC-CMs. After optimizing experimental perturbations at these two critical developmental stages, we will employ a combinatorial strategy to enhance the consistency of high purity hPSC-CM differentiation by merging the signaling approaches utilized to generate high potency mesoderm and cardiac progenitors. In summary, the proposed work drives progress towards improved heart disease treatment strategies by temporally investigating high and low purity hPSC-CM differentiations to unveil the molecular mechanisms of human cardiogenesis and to facilitate robust, high purity hPSC-CM biomanufacturing. The proposed work will utilize the stem cell bioengineering, cardiomyocyte biology, and single nucleus multi-omics expertise of the Sponsor, several collaborators, and outstanding research cores at the University of Wisconsin to investigate and control how human cardiogenesis unfolds in vitro. The training opportunities afforded by this proposal will facilitate extraordinary research, clinical, professional, and leadership growth, which are all essential skills to develop into a successful physician scientist.

NIH Research Projects · FY 2025 · 2024-09

Abstract Mitochondria, the energy generating organelles, dynamically fuse and fission to take various forms in different cells. Mitochondrial dynamics have been linked to complex cellular processes such as metabolism, immune response, and cell death, but the exact relations between their form and function are still not fully understood. Retinal rod photoreceptor cells, the energy intensive neurons, provide an excellent model for investigating the significance of mitochondrial form because they have a uniquely uniform arrangement of elongated mitochondria in the inner segments and one large circular mitochondrion in each of the presynaptic terminals. In our preliminary study, rod photoreceptor cell-specific ablation of mitochondrial fusion proteins, mitofusin 1 (MFN1) and mitofusin 2 (MFN2), resulted in fragmentation of mitochondria by one month of age, followed by extensive photoreceptor cell degeneration by the adult age. In addition, rod photoreceptor cell- specific ablation of a mitochondrial fission factor, dynamin-related protein 1 (DRP1) caused late-onset and age- dependent retinal degeneration (obvious by one year of age), suggesting the role of DRP1 in aging rod photoreceptor cells. Therefore, we hypothesize that development and maintenance of the mitochondrial form through mitochondrial dynamics are vital for sustaining rod photoreceptor cell health and function. In Aim 1 of this proposal, we will test the hypothesis that mitochondrial fusion is critical for establishing rod photoreceptor cell-specific mitochondrial structures and metabolic status during postnatal development. In Aim 2, we will test the hypothesis that mitofusins maintain the health of rod photoreceptor cells by affecting the metabolic pathways. In Aim 3, we will examine the role of the mitochondrial quality control mechanism through DRP1 in aging rod photoreceptor cells. Successful completion of these studies will provide novel insights into the functional significance of the regulation of mitochondrial dynamics in rod photoreceptor cells, downstream effects of changes in the mitochondrial form, and the link between mitochondrial dynamics dysfunction and neurodegeneration.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Women and birthing people who identify as African American are more likely to experience severe maternal morbidity or mortality or deliver a low-birth weight infant and experience hypertensive disorders of pregnancy at higher rates. Factors leading to health disparities around chronic disease such as hypertension, cardiovascular disease, and diabetes are set early in life: the long-term effects of health disparities in pregnancy and early childhood signal that interventions at this time of life may have a positive impact on adult health and chronic disease. The lived environment and socioeconomic status have a larger impact on outcomes than health behaviors and clinical care, so advancing effective prevention strategies that eliminate deleterious social determinants of health is an urgent priority. The overarching mission of the University of Wisconsin-Madison Prevention Research Center (UWPRC) is to improve the health of women and birthing people, infants, and families impacted by health inequities, by conducting and building capacity for high-quality applied health promotion and disease prevention research. The proposed UWPRC will build on and extend the success of the existing UWPRC. UWPRC will be led by an experienced Administrative Team who engage with a broad base of academic experts through a Steering Committee and Faculty Advisors Committee. Community consultation and engagement through the Community Advisory Board and the Translational Partners Panel, enable high-quality and community-grounded intervention, implementation, and public health practice-based research and translation. The UWPRC will be housed in the School of Medicine and Public Health, a medical school that integrates public health in its research, teaching, and service missions and is integral to Wisconsin’s public health system. The core research project, “Community-based health promotion and prevention modifying Staying Healthy After (Around) Childbirth (STAC) for birthing persons at risk for and with hypertension during pregnancy through postpartum” is a dissemination and implementation project to adapt STAC, an evidence- based intervention for remote blood pressure monitoring for hypertension disorders in pregnancy (chronic disease priority category 5). The project goals are to initiate remote hypertension care during pregnancy and through six-weeks postpartum and incorporate implementation of the program in community-based organizations through a community-engaged participatory approach to improve identification, prompt treatment, in Black persons at risk for or with hypertensive disorders in pregnancy. The project will build relationships with community-based organizations led by Black women and those serving Black women, and address gaps in scalability and identification of necessary supports for adoption and implementation of the intervention. The UWPRC will continue and expand its participation in the PRC Network throughout the funding cycle. We are uniquely positioned to develop, test, and translate evidence-based strategies at the interface of communities, public health, and health care, where translational barriers often delay or prevent their impact.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Central nervous system (CNS) neurons exhibit a limited capacity to survive and regenerate axons following injury, resulting in functional failure in the brain (e.g., stroke or trauma), spinal cord (e.g., paralysis), or the optic pathway (e.g., blindness). However, 5 to 10% of CNS neurons do survive injury and only 10 to 15% of these surviving CNS neurons regrow axons. Critical to understanding the molecular mechanisms of neuronal survival and regeneration is isolating regenerating from non-regenerating neurons and deciphering their unique gene expression and regulatory profiles following injury. We developed a unique method of purifying regenerated from non-regenerated retinal ganglion cells (RGCs) following injury and identified, using bulk whole-genome methylation sequencing, differentially methylated regions throughout the rat genome capable of distinguishing regenerated from non-regenerated RGCs. Bulk RNA-sequencing of these same two RGC populations revealed differentially expressed genes similarly capable of distinguishing regenerating from non-regenerating RGCs, and some of these differentially expressed genes were also differentially methylated. DNA methylation levels are established and maintained by the folate biochemical pathway; thus, discerning specific DNA methylation alterations that correlate with RGC regeneration supports our previous observations that folic acid supplementation improves the ability of RGCs to regenerate axons following injury. Recent reports showed that 46 different RGC subtypes exist and some of which exhibit particular resilience or susceptibility to neurodegeneration. These findings suggest that select RGC subtypes contribute to axon regeneration. Accordingly, using our published optic nerve transection model and established high-throughput single-cell methylomic and transcriptomic protocols, we will determine the molecular profiles of purified regenerating RGCs to identify specific RGC subtypes that contribute to axon regeneration. Moreover, we hypothesize that folic acid alters select regenerating and non-regenerating RGC subtypes to enhance folic acid-dependent axon regeneration. An improved understanding of how different RGC subtypes contribute to axon regeneration can be leveraged to reveal novel neuroprotective mechanisms for a vista of new research opportunities across various neurological disorders of the brain, spinal cord, and optic pathway.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy) is the most common monogenic cause of stroke and vascular contributions to cognitive impairment and dementia (VCID). In this project, we will study CADASIL patients with NOTCH3 mutations. Preclinical studies have indicated that NOTCH3 mutations are associated with structural and functional changes in the vascular smooth muscle cells and the loss of pericyte function. Thus, studying CADASIL patients with NOTCH3 mutations is an ideal approach to investigate early physiological changes associated with VCID in gene mutation carriers, prior to the onset of clinical symptoms. We will examine cerebrovascular reactivity (CVR), a cerebral small vessel disease (SVD) functional marker, in a subset of participants from the first North American longitudinal cohort (CADASIL Consortium). This CADASIL Consortium includes 400 adults with CADASIL NOTCH3 mutations and 100 non-carrier controls. A key feature of CADASIL and other SVD is the early development of white matter (WM) hyperintensities and these features are often irreversible. Functional declines in microvascular health are thought to occur earlier in the development of the disease and may underlie the WM changes in CADASIL. CVR is a sensitive measure of microvascular function and reflects the ability of small vessels to dilate in response to vasoactive stimuli. Thus, the objective of this application is to determine the impact of NOTCH3 mutations on CVR, and explore potential associations with clinical, neuropsychological, neuroimaging, and biofluid markers of VCID. To achieve this, we will leverage participants and data from the longitudinal cohort with NOTCH3 mutation carriers (CADASIL Consortium) and compare with healthy non-carriers. To enhance feasibility, we will use a novel, yet validated CVR assessment that uses breath modulation and is suitable for multi-site MRI data collection in the following specific aims. In Aim 1 we will examine if CVR is lower in NOTCH3 mutation carriers with varying disease severity (asymptomatic, symptomatic, and symptomatic with functional decline) when compared with healthy non-carrier controls. In Aim 2 we will determine if CVR in NOTCH3 mutation carriers is associated with other biomarkers of VCID and CADASIL phenotypic markers. In Aim 3 we will characterize the trajectory of change in CVR in NOTCH3 mutation carriers and non-carrier controls (over 18 months) and determine the association with other biomarkers of VCID and CADASIL phenotypic markers. Upon completion, this application will provide critical information on the pathogenesis of VCID using participants with a SVD autosomal dominant source, that will expand beyond patients of the rare disease CADASIL. This approach aligns with recent NIH recommendations emphasizing the need for human studies to identify and confirm biomarkers of vascular processes related to cognitive impairment. Importantly, this effort is timely and complements ongoing NIH-funded work to understand Alzheimer’s Disease Related Dementias, including the multi-site MarkVCID study, in which CVR is being evaluated as a candidate biomarker for sporadic VCID.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Synaptic loss is theorized to occur early in the course of common neurodegenerative proteinopathies such as Alzheimer disease (AD) and Lewy body dementia (LBD). Until recently, the evaluation of synaptic loss in the human brain depended on the availability of pathologic/autopsy material, which inevitably did not represent early stages of disease. In the past few years, development of radiopharmaceuticals that target SV2A, a ubiquitous synaptic protein, have made it possible to quantify synaptic density in the human brain in vivo. One long-term goal of the collaborative neuroimaging programs at UW-Madison is to use magnetic resonance imaging (MRI) and Positron Emission Tomography (PET) to understand and detect preclinical stages of neurodegeneration in AD and ADRD. The objective of this proposal is to use SV2A PET, acquired simultaneously with diffusion and structural MRI, to evaluate the sequence and topography of synaptic, white matter microstructural, and volumetric changes that correspond to prodromal and manifest stages of LBD. The central hypothesis is that lower synaptic density in neocortex and hippocampus will be detectable at earlier disease stages than reductions in white matter microstructural integrity and will correspond to or predict cognitive decline. The rationale for this proposal is that participants selected and carefully characterized to represent the spectrum of disease stages from healthy aging to prodromal LBD will provide a representative cohort from which to draw conclusions regarding the sequence of brain changes that occur prior to clinical disease onset. For this project we will recruit 160 human research participants, 120 of whom have significant risk factors to develop LBD: Forty participants with rapid eye movement sleep behavior disorder and normal cognition (RBD-NC), 40 participants with early- stage Parkinsonism and normal cognition (PD-NC), and 40 participants who meet research criteria for mild cognitive impairment with Lewy body (MCI-LB), as well as 40 age- and sex-matched healthy controls. We are experienced in and will perform extensive motor and cognitive characterization of these individuals at baseline and 2-year follow-up intervals. All participants will undergo structural and diffusion MRI as well as SV2A PET at baseline; a subset of participants (~35) who show cognitive decline during the study will be re-imaged with SV2A PET/MRI at follow-up. Using the SV2A PET and MRI data, the specific aims will be to (1) map differences in synaptic density between control, RBD-NC, PD-NC, and MCI-LB at baseline, (2) determine the relationship between synaptic density within prespecified neo and allocortical regions and cognitive decline, and (3) determine the degree to which reduced synaptic density occurs independent of microstructural and structural change prior to dementia. The significance of this project is that it evaluates important hypotheses regarding the role of synaptic loss as an early event in and cause of cognitive change in LBD. This project is innovative because few studies have evaluated SV2A in prodromal stages of LBD, including RBD and MCI-LB. The positive impact of this work will be to develop biomarkers for the future evaluation of intervention therapies.

NIH Research Projects · FY 2025 · 2024-09

Abstract With nearly 40 years of experience, the Wisconsin Research and Education Network (WREN) is one of the oldest and most respected primary care practice-based research networks (PBRNs) in the US, having been founded in 1987. WREN has conducted hundreds of clinical trials in primary care practices, including pragmatic trials, implementation science studies, and community-engaged projects, among others. Given our extensive experience, deep expertise, and strong relationships with rural primary care clinics, WREN is well poised to serve as a Rural Network Research Hub. WREN is supported by the University of Wisconsin-Madison (UW-Madison) School of Medicine and Public Health CTSA (Institute for Clinical and Translational Research, ICTR) and is in the UW-Madison Department of Family Medicine and Community Health. WREN is in the Meta-network Learning and Research Center (Meta- LARC), a bi-national network of PBRNs. As a Rural Network Research Hub, WREN will partner with two rural Federally Qualified Health Centers (each with multiple clinics) serving Western and Northern Wisconsin, an independent practice serving Southwestern Wisconsin, and UW Health rural clinics. These clinics have successfully worked with WREN on a variety of projects. In year 2, we will expand our network to new rural clinics and re-engage with clinics we worked with previously. WREN will facilitate regular meetings of the Hub clinics in a learning collaborative to assess, select, and facilitate NIH study opportunities in their rural communities. We will develop infrastructure to support rural clinics in implementing research and build and sustain engagement with rural communities underrepresented in clinical research.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Human cytomegalovirus (HCMV), like all herpesviruses, has two phases of its infectious cycle that enable lifelong infection in its host. Lytic infection occurs in differentiated cells and produces infectious virions. Latent infection occurs in undifferentiated cells and is characterized by limited viral gene expression, lack of virion production, and the potential for reactivation. HCMV virions have a tegument layer around the capsid containing proteins important for initial events in infection. In differentiated cells, tegument-delivered viral transactivator pp71 traffics to the nucleus, degrades the cellular repressor DAXX, activates lytic immediate early (IE) genes, and lytic infection is initiated. In undifferentiated cells, tegument-delivered pp71 does not reach the nucleus, DAXX is not degraded, IE transcription is silenced, and latent infection is established. This is because in undifferentiated cells, pp71 and some other tegument proteins remain colocalized with endosomes, while the genome-containing capsid and capsid-associated tegument proteins reach the nucleus. However, the mechanism leading to tegument disassembly and the nuclear localization of tegument-delivered pp71 in differentiated cells and why it does not occur in undifferentiated cells is unknown. Because HCMV enters fibroblasts (where tegument-delivered pp71 reaches the nucleus) by virion fusion at the plasma membrane, but enters myeloid cells (where tegument-delivered pp71 remains cytoplasmic) by endocytosis, I hypothesize that entry by fusion drives pp71 nuclear localization. Here I will test this hypothesis by chemically and biologically inducing HCMV to fuse into myeloid cells and then monitor tegument-delivered pp71 subcellular localization, IE transcription, and infection outcome.

NIH Research Projects · FY 2024 · 2024-09

Economic activity across sectors can impact human health through multiple exposure pathways. Many health risks are well-documented, for example, the risk of air pollution on health outcomes across the lifespan. Major policies, however, continue to emerge without a full consideration of their health implications. Lacking is an understanding of how policy decision making (outside of the health sector) can have specific and measurable public health implications. To fill this gap, our proposed Center for Health, Energy, and Environmental Research (CHEER) will catalyze rigorous, innovative, community-driven, policy-relevant, transdisciplinary research. Our overarching aim is to better understand the positive and negative health impacts of cross-sector policy choices across energy, transportation, land use, and infrastructure.  Achieving this goal requires engagement across community, scientific, and policy sectors. We posit that a truly community engaged approach to decision-making will increase both the efficacy and durability associated with specific policy solution pathways proposed with human health in mind. Our key innovation is in developing and employing a linked pathway that starts and ends with communities while it generates robust estimates of the health impacts of contemplated cross-sector policies. Innovations in community engagement will include: participatory web-based technologies, including web-based mapping and storytelling, crowd-sourcing and artificial intelligence, and utilization of low-cost air pollution monitors, which are increasingly being used by citizen scientists and communities. Innovations coming from our primary research project will include development of a new, high-resolution Scenario Health Risk for Energy model to support decision-making by policy makers and community partners, balancing representation of complex processes with the speed and ease-of-use required for public utilization. We also will conduct a Pilot Target Trial to assess the likely impacts of variable air pollution scenarios on asthma burden of Milwaukee public school children. The proposed Data Science Core will prioritize authentic, multi-directional, equitable community engagement and data sharing. Innovations will stem from: Establishing a Data Repository that will foster team science and provide a single point of access for all stakeholders; creating a Data Commons that provides linkage, integration, mapping, metadata management, and computerized routines for reusable data, metadata, and software artifacts for collaborative research and data sharing; and Providing Digital Engagement tools using advanced technologies (e.g., natural language processing) to ensure that community voices are utilized efficiently to inform research and policy development. Our Wisconsin-based community, government, and university environments have the track record, expertise, foundations, and institutional support for high productivity and policy impact of this Center for Health, Energy, and Environmental Research.

NIH Research Projects · FY 2025 · 2024-09

Abstract Urinary urgency, with or without incontinence, greatly affects the quality of life in individuals suffering from Overactive Bladder (OAB). A sudden and often overwhelming desire to pass urine occurs frequently (8 times and more) throughout the day, often coinciding with nocturia. More prevalent in females, the overall prevalence in U.S. studies is 14%. Lifestyle modifications, bladder retraining, pelvic floor exercise and antimuscarinic OAB therapies are initial treatments for OAB. Although these can provide some relief, complete relief is rare, and treatment is entirely unsuccessful in 25-40% of women (refractory OAB) due to lack of initial effect, developed tolerance, or intolerable side effects leading to discontinuation. Both implantable and non-invasive posterior tibial nerve stimulation (PTNS) devices have been FDA approved to treat refractory OAB. These existing PTNS devices are placed many millimeters or even centimeters from the nerve, and have significant issues with inconsistent targeting/migration, with stimulation induced side-effects putatively limiting engagement with intended A-beta sensory fibers within the tibial nerve. Implantable PTNS devices are also costly and complex, leading to significant device related complications. To address these issues, we will use a rational engineering approach to develop, validate and optimize a novel minimally invasive electrode strategy consisting of only 2 implanted passive materials, and an associated surgical tool that allows for direct targeting of the nerve via injection. We anticipate this will yield dramatically improved nerve engagement while reducing complications.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT Beginning early in life, metabolic dysregulation related to the triplication of chromosome 21(e.g. decreased energy expenditure, increased leptin, hypotonia, increased inflammation, and autonomic dysfunction) as well as lifestyle (e.g., poor diet, sedentary behavior, reduced and disrupted sleep, and stressors) are thought to contribute to the 2 times greater prevalence of obesity in people with Down syndrome (DS) as compared to the general population. In the general population, metabolic health, obesity, and lifestyle have strong associations with health conditions including obstructive sleep apnea, Alzheimer’s disease, cardiometabolic diseases, and cognitive impairments. However, the relationship between these factors in people with DS is not well understood. The Metabolic Health, Lifestyle, and Risk of Co-Occurring Health Conditions in Down Syndrome (MET-DS) study is a five-year, longitudinal study of factors that alter the risk and severity of co-occurring health conditions in children, adolescents, and young adults with DS. The study involves a rigorous deep-phenotyping protocol to better understand the complex interplay between trisomy 21, metabolic dysregulation, obesity, lifestyle, and the development of co-occurring health conditions. This effort will enroll 200 participants (ages 6- 24 years of age) with DS from four clinical performance sites, and follow them annually across 3 data collection cycles to address the follow aims: 1) Deeply characterize and establish normative data on metabolic health, obesity, and lifestyle factors of a large cohort (N = 200) of children, adolescents, and young adults with DS (aged 6-24 yrs.); 2) Measure the stability and change in profiles of metabolic health, weight status, and lifestyle factors across 2 years (3 time points; 12 months apart) and during transitional periods of development; 3) Establish the relation between metabolic health, obesity, and lifestyle factors on co-occurring health conditions (e.g., obstructive sleep apnea, cardiometabolic diseases, autoimmune diseases, and early biomarkers of Alzheimer’s disease) and cognitive development across 2 years (3 time points; 12 months apart); 4) Collaborate with the other DS-CRS sites and the DS-4C on the development, implementation, and data harmonization of common core measures across domains (e.g., clinical, cognitive, behavioral, imaging, and multi-omics) and sites, and lead the DS-CRS sites in collection of metabolic and lifestyle measures.

NIH Research Projects · FY 2026 · 2024-09

ABSTRACT Intergenerational transmission of inequalities in economic and social wellbeing, including health, in the United States has been well documented and shown to account for a substantial portion of contemporary disparities by SES and race and ethnicity in these domains. Yet, due to a paucity of population-based longitudinal data on family contexts, parenting, and child development across multiple generations, knowledge of the extent to which these factors may be key mechanisms for intergenerational transmission of inequalities is limited. The Future of (formerly Fragile) Families and Child Wellbeing Study (FF) is the only ongoing population-based birth cohort study in the U.S. to currently follow three generations of family members. FF follows the parents (first generation; G1) of initial birth cohort children (G2; born 1998-2000) and the G2 children (now young adults). Survey and biological data are now being collected on all children born to G2 sample members (G3) as soon as possible after their birth and will be collected on G2s at age 27. Data span economic, social, environmental, and biological factors for each generation within a large and diverse national sample. The proposed study will expand FF to include data at age 5 for all (G3) children born to G2 women. The project aims to: (1) Collect data on family/caregiving context, parenting and grandparenting behaviors, and child health, socioemotional development, achievement (cognition), and executive function for all G3 children born to female G2 respondents as they reach age 5; (2) Prepare all data and accompanying documentation to be publicly available to the research community and fully linkable to all other FF data sources within 1 year of the end of the award period; (3) Produce a comprehensive guide for implementing virtual assessments of children and families in the context of a large-scale national study with specific attention to logistical considerations, response rates, potential challenges (connectivity and quality therein, household distractions, child attention and comprehension), and lessons learned, both overall and for particular subgroups of families; and (4) Conduct preliminary analyses of intergenerational correlations of family context/caregiving environment, economic resources, parenting behaviors, and child development among disadvantaged families. These data will support novel and important studies of intergenerational transmission of family context, economic resources, parenting behaviors, and child development; whether associations of family context, economic resources, and parenting behaviors with child development are consistent or differ across generations; and whether there is heterogeneity in these patterns by population characteristics. They will also (eventually) support epigenetic analyses via DNA-based, sociodemographic, and environmental information on three generations, including prenatal, perinatal, and early childhood data on G2 and G3. As such, FF, with the inclusion of G3 early childhood assessments, holds unlimited potential to support novel and innovative multi- generational research to inform public health and social policy.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY The introduction of highly pathogenic avian influenza (HPAI) virus of the Guangdong H5N1 lineage has resulted in severe disease outbreaks with widespread mortality in wild birds and poultry in the United States. Current control measures, including massive culling of infected flocks, vaccination efforts, and biosecurity procedures have failed to stop the spread of avian viruses including HPAI viruses resulting in significant economic losses and public health risks due to their zoonotic potential. There is a pressing need to explore alternative strategies to combat viral pathogens that inflict massive losses on the poultry industry. To tackle the susceptibility of chickens to avian viruses, we propose establishing the BRAVE (Building Resistance Against Viral Entities) Innovation Center. This initiative aims to generate chickens with intentional genome alterations that are resistant to viral pathogens of significant biological and economic importance. To identify the host factors that will be targeted for intentional genome alterations in chickens, we will perform a genome-wide CRISPR/Cas9 study with a sgRNA library in Aim 1. These experiments will focus on identifying pro-viral chicken genes that are essential for virus replication. We will begin with HPAI H5N1 virus, and then expand the study to HPAI H7N9 virus and other avian viruses including infectious bronchitis virus (IBV). Aim 1 will also identify host factor targets for intentional genome alterations to identify unique and common antiviral proteins that directly inhibit the replication of the viruses in this proposal. We will identify and rank the best host factors when knocked out (pro-viral factors) or overexpressed (antiviral factors) for the greatest inhibitory effect on virus replication. We will monitor cell viability and the lack of escape viruses that would mutate away from the resistance profile. Using the data collected in Aim 1, in Aim 2, we will generate an intentional genomically altered (IGA) chicken line resistant to HPAI virus infection and second line that has a multi-virus resistance phenotype. We will employ CRISPR/Cas9 or transposase methods to modify the genomes in chicken primordial germ cells (PGCs) and then inject the modified PGCs into recipient chicken embryos to establish germline-modified chickens and eventually flocks of birds for infection studies in Aim 3. Once an IGA chicken line is established, we will compare the susceptibility of the altered birds with that of wild-type birds to HPAI H5N1 and H7N9 virus infections. Virus replication, associated pathology, transmission, and emergence of mutant viruses associated with breakthrough infections will be assessed and compared to wild-type birds. We will also evaluate the susceptibility of the multi- virus-resistant chicken line to H5N1 and H7N9 viruses, along with other avian viruses including IBV. Non-infected IGA chicken lines will be characterized to ensure proper growth, longevity, and fertility along with any pathological changes compared to their wild-type counterparts.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY: Hematopoietic stem cells (HSCs) are at the top of a hierarchy of differentiating progenitor cells that can reconstitute the entire blood and immune system. Proper regulation of HSC differentiation is critical during homeostasis and response to stress or infection. Dysregulation of HSCs naturally occurs during aging, and aberrant differentiation can cause blood cancers and other diseases. Decades of research have been devoted to understanding and defining the hierarchy of HSC differentiation. However, the limitations of current biological tools and animal models has left key questions unanswered. Foremost, we have not been able to track individual HSCs and multipotent progenitors (MPPs), directly and continuously, in their endogenous microenvironment. Previously, HSCs and MPPs in mouse models could only be identified by dissociation of bone marrow, labeling with antibodies to detect markers of differentiated lineages (Lin) and at least four other markers; for example, c-Kit (aka Kit or CD117), Sca-1 (aka Ly6a), CD48, CD150 (aka Slamf1). More recently, single color transgenic HSC reporter lines have labeled a proportion of HSCs that could be detected by intravital imaging, but without additional markers for MPPs, the dynamics of HSC fate decisions are lost. We hypothesize that newly discovered combinations of cell type-specific markers will allow live tracking of HSC/MPP behaviors and key lineage decisions. The convergence of multiple cutting-edge technologies makes it possible to address this hypothesis by generating a multi-color, multi-loci mouse reporter system in the following aims. Aim 1) Generate an HSC/MPP reporter mouse by targeting knock-in 2A fusion reporters to four loci. We will use CRISPR/Cas9-dependent homology directed repair (HDR) in mouse zygotes, with repair template delivered by adeno-associated virus (AAV). Each locus will retain functional proteins that cleave to express distinct fluorescent proteins (e.g., BFP, GFP, tdTomato, iRFP), representing a positive or negative marker for HSCs and/or MPPs. Aim 2) Characterize the HSC/MPP reporter line using complementary assays to assess faithful labeling of HSCs and MPPs. After knock-in validation, we will interbreed the lines to generate four color transgenics. Labeled HSCs and MPPs will be quantified by flow cytometry and functionally validated by limiting dilution transplantation. Bones will be imaged ex vivo to confirm that spatial maps of endogenously labeled HSCs and MPPs are consistent with published immunolabeling data. Outstanding questions about the migration of HSCs and MPPs will be answered by intravital microscopy. Our research proposal fits this funding opportunity because it represents a significant “improvement of animal models for stem cell-based regenerative medicine.” It addresses the research interests of multiple NIH Institutes and Centers because the models generated would apply to the basic biology of the blood system (NHLBI), immune cells (NIAID), aging hematopoiesis (NIA), HSC biology (NIDDK), and blood cancers (NCI). As blood and immune cells interact with all tissues of the body, our models will be used to evaluate “processes that impact multiple body or organ systems.”

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT: The goal of this study is to characterize a novel mouse mutant with a phenotype resembling Braak stage I/II Alzheimer’s disease (AD) pathology in humans and to develop this line into a novel animal model for AD research. AD is a major public health crisis predicted to affect 13.8 million Americans and cost $750 billion per year by 2050. However, the pathological mechanisms underlying AD have remained controversial, especially regarding how amyloidosis, tauopathy, and neuroinflammation, the three hallmarks of AD, may interact with each other at the molecular and cellular level to promote AD pathology development. This uncertainty has impeded the development of effective intervention. In pilot studies, we have generated a microglia-specific mouse mutant with strong tauopathy and atrophy specifically in the entorhinal cortex. This is the first time such a unique tauopathy phenotype has been observed in the mouse brain. In humans, tauopathy in the entorhinal cortex predicts not only that rate of future tau pathology spread and deterioration in the brain but also the rate of cognitive decline in affected patients. Thus, the mouse mutant we have provides a unique opportunity for uncovering mechanisms of tauopathy development in AD. To characterize this mutant, we will: 1) determine spatial and temporal pathologic progression across cell types in the mutant brain 2) identify key cellular/molecular drivers of the tauopathy phenotype Through these experiments, we will determine primary and secondary changes in the various brain cell types in the mutant brain and pave way for elucidation of mechanisms of AD tauopathy development in this unique mouse model. We will also gain insight into the mechanisms by which inflammatory cytokines may drive entorhinal tauopathy in this mutant and pave way for determining the functional significance. Together, through this study, we will provide not only a novel AD animal model with strong entorhinal tauopathy and neurodegeneration yet no complications of gene overexpression. Furthermore, since the predominant group of risk genes for late-onset AD, which comprises >95% of AD cases, are related to microglial function, we will also provide a model to elucidate pathogenic mechanisms of tauopathy that are specific and relevant to the vast majorities of AD cases.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT / PROJECT SUMMARY The prevalence of combustible cigarette tobacco use remains high among socioeconomically disadvantaged Americans despite declines in the broader population. Socioeconomic inequities in tobacco use rates and tobacco cessation success have grown over recent decades. Disadvantaged adults who use tobacco are therefore a high-priority population, but we have made too little progress in engaging this population in evidence-based tobacco cessation treatment. Community agencies that provide support services to disadvantaged individuals have the potential to reach people who use tobacco and engage them in high-quality cessation treatment to prevent cancer. The Salvation Army is a community agency with national and international reach and a commitment to discouraging tobacco use and promoting the health and wellbeing of the diverse communities they serve. The Salvation Army will partner in the proposed project to refine and evaluate implementation strategies designed to help their staff connect their clients with free evidence-based tobacco treatment from the Wisconsin Tobacco Quit Line. Salvation Army social service leaders, staff, and clients will help to refine and adapt quitline referral implementation strategies with promising preliminary data to enhance their fit and sustainability in the usual context of Salvation Army services. Pragmatic research methods will be used to evaluate two promising implementation strategies designed to promote quitline referrals: 1) a low-intensity, sustained collaborative approach to implementation support for Salvation Army staff (vs. a usual care approach offering initial training with minimal technical support), and 2) modest, immediate incentives for clients to connect with the quitline (vs. no incentive). Both the collaborative sustained implementation support for staff and client incentives will be refined in the 2-year UG3 phase, along with pragmatic study measures and procedures. In the UH3 phase, the target implementation strategies refined in the pilot phase will then be evaluated in a pragmatic randomized rollout trial in which 8 sites are randomized to sustained collaborative implementation support for staff and 8 sites are randomized to usual implementation training for staff, and all 16 sites will start without client incentives for immediate quitline connections, and will then introduce incentives at randomly assigned timepoints, as in a stepped-wedge design. Inclusion criteria for sites, staff, and clients will be minimal to enhance external validity, and data on the primary outcome (whether or not a referral occurred) will be gathered from Salvation Army service records and quitline records in the course of usual care. This will be supplemented by mixed method assessments conducted by researchers that will not affect the context of intervention implementation. This pragmatic program of research has the potential to identify staff- and client-focused strategies that can enhance Salvation Army efforts to support tobacco cessation among their clients. Leaders at the Salvation Army are committed to helping their clients quit tobacco, and to applying the lessons learned in the proposed project more broadly in the region and nation.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY Where one lives is an important contributor to disparities in health behaviors and outcomes, but it has been difficult to draw firm conclusions on the relations between regional/neighborhood context and alcohol use and misuse. This may be because (a) the influence of alcohol use and misuse (and genetic risk for alcohol use and misuse) on where one lives is rarely considered, and (b) the effects of regional/neighborhood contexts may only be relevant for certain genetically vulnerable individuals. Incorporating genetic information into a prospective longitudinal study of the relation between regional/neighborhood context and alcohol use and misuse might resolve inconsistencies in the literature. The proposed research will involve secondary analyses of data from the National Longitudinal Study of Adolescent to Adult Health (AH), which included 20,745 adolescents in 1994-95 and has since conducted four follow-up waves. All five waves of data collection gathered extensive health and behavior information, including alcohol use and misuse; administrative data from the census and other sources were linked to the participants’ home addresses to provide information about the region/neighborhoods in which they lived. Importantly, 9,974 participants provided DNA samples for genotyping, allowing for the quantification of alcohol-related genetic risk by aggregating the effect of individual measured polymorphisms into polygenic risk scores. The inclusion of 1,962 (289 monozygotic twin, 452 dizygotic twin, and 1,251 full sibling pairs) in the sample allows for the control of genetic factors when examining associations between alcohol involvement and regional/neighborhood context and can provide evidence consistent with a potentially causal relation. The three main goals of this proposal are to: (1) examine prospectively the relation between regional/neighborhood contexts and later alcohol use and misuse, and (in adulthood) the prospective relation between alcohol use and misuse and later regional/neighborhood contexts, (2) examine prospectively, within exposure-discordant adult twin and sibling pairs, the relation between regional/neighborhood contexts and later alcohol use and misuse, and the prospective relation between alcohol use and misuse and later regional/neighborhood contexts, (3) examine whether alcohol-related polygenic risk is associated with moving to or remaining in a high-risk neighborhood context and whether alcohol-related polygenic risk amplifies the associations between neighborhood context and alcohol use and misuse. The proposed research has the potential to provide critical insights into how alcohol-related genetic propensities may exacerbate health disparities by influencing whether individuals reside in a high-risk neighborhood context. It will also inform prevention and intervention efforts by clarifying whether the relation between neighborhood context and alcohol misuse involves a direct causal relation. Evidence consistent with such a causal relation would point to neighborhood-level interventions; evidence of a non-causal relation would suggest that interventions delivered directly to individuals or families might prove more effective.

NSF Awards · FY 2024 · 2024-09

Nuclear quantum effects significantly impact the static and dynamic properties of many chemical, biological, and materials systems. Accurately and efficiently incorporating nuclear quantum effects in quantum chemistry calculations and molecular dynamics simulations is a crucial challenge in the field, especially for systems with light nuclei like hydrogen. The constrained nuclear-electronic orbital density functional theory (CNEO-DFT) and its molecular dynamics extension (CNEO-MD), developed by the Yang group, provide high accuracy and efficiency in describing these effects. The growing interest in utilizing these methods for various investigations necessitates the development of robust, reliable, and sustainable cyberinfrastructure products. The project aims to develop cyberinfrastructure products that integrate constrained nuclear-electronic orbital functionalities into widely used software packages. This open-source cyberinfrastructure ensures wide accessibility for the quantum chemistry and molecular dynamics communities and fosters future research in theoretical and computational chemistry. Additionally, it has broader impacts on drug discovery and education in physical and theoretical chemistry. The project aligns with NSF’s mission to advance scientific progress and has the potential to significantly impact multiple research fields by providing a powerful computational tool for accurate simulations of systems with significant nuclear quantum effects. The technical scope involves integrating CNEO-DFT and CNEO-MD functionalities into existing open-source community-based packages. Specifically, these functionalities will be implemented as modular extensions in PySCF, in NWChem for practical and large-scale simulations, in codes interfacing PySCF extensions with GROMACS for hybrid quantum mechanics/molecular mechanics (QM/MM) calculations, and in CP2K for condensed-phase simulations. Each developed element primarily targets a community that includes quantum chemistry method developers, computational and experimental researchers in gas-phase chemistry, computational biochemists, and chemists as well as materials scientists in the condensed-phase community. This award by the NSF Office of Advanced Cyberinfrastructure is jointly supported by the Division of Chemistry. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Toxic oligomers formed by amyloid-beta (Aβ) self-association are of high interest for research into the causes and treatment of Alzheimer's Disease. Studies Aβ oligomer formation on a molecular level using biophysical and analytical techniques have provided many valuable insights, but there remain many open questions regarding oligomer structure and the factors that influence their formation. In the present proposal, we will develop a new method for studying Aβ oligomerization that greatly diverges from the biochemical and biophysical approaches historically utilized by the field. Here, we will use a genetically encoded sensor that detects Aβ oligomer formation and converts the oligomer formation event to the expression of a reporter gene, enabling high-throughput and direct measurements of Aβ oligomerization. This format enables us to measure the effects of millions of different factors on Aβ oligomerization and to quickly identify agents that can selectively bind oligomers over other Aβ species. 1

NIH Research Projects · FY 2026 · 2024-09

Project Summary/Abstract Kinship ties form the bedrock of societies and provide meaning in terms of roles, obligations, and responsi- bilities. Generational overlap in the form of shared lifetimes represents a fundamental condition guiding whether and how kin relationships may develop and the extent to which resources are shared. Yet, we know little about the prevalence, duration, or life course timing of generational overlap, how it varies by socioeco- nomic status, and how it may potentially influence individual health and wellbeing. Generational overlap is con- sequential because it offers an important perspective for observing the demographic constraints on multigener- ational family investments in children. We propose to provide new information about the overlap in shared life- times for grandchildren and grandparents across two industrialized countries – the U.S. and Denmark. Both countries have experienced declines and delays in fertility, lengthening life expectancies, and notable educa- tional expansion in the 20th century, but these have occurred on different timelines, to different degrees, and within different social policy regimes. We use harmonized data from the Panel Study of Income Dynamics, the Danish population register, the Add Health Parent Study and the Survey of Health, Aging, and Retirement in Europe to evaluate generational overlap and its implications across historical time. We consider three genera- tions – grandparents (G1), parents (G2), and grandchildren (G3) and focus on grandparents born since 1912. We address three specific aims: 1) Provide a descriptive portrait of the prevalence and length of generational overlap for grandparents (G1) and grandchildren (G3) in the U.S. and Denmark using an innovative age-pe- riod-cohort approach; 2) Explicate the role of educational expansion in shifting generational overlap, including the specific case of within-family intergenerational mobility where diminished generational overlap may be an unanticipated `cost' of intergenerational mobility; 3) Document the social class gradient in grandparents' life course position (employment status, health, and proximity to kin) arising from the timing and duration of gener- ational overlap and describe the intergenerational exchanges of time, money, and care that result – and for Denmark, consider subsequent health/wellbeing for G1-G3. We attend to further variation by gender, family structure, and (for the U.S.) race/ethnicity. In sum, shared lifetimes among grandparents, parents and grand- children represent a key aspect of how kinship ties shape individual life courses. We explore how the duration, quality, and consequences of these shared lifetimes are shaped by demographic processes across genera- tions. This project will be led by an outstanding team of established collaborators who are leaders in the fields of family demography and social inequality and who possess the methodological skill and expert knowledge of each data source to achieve the study's aims. These findings have important implications for the intergenera- tional transmission of inequality, as well as potential resource demands on governments and families.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT Mammals have evolved an extensive nuclear network of DNA replication and repair proteins that are tasked with amplifying and maintaining the fidelity of their genetic code. These signaling mechanisms are always active, surveying the genome to maintain its integrity in the face of attack from external abiotic factors (such as radiation and chemicals) and biotic factors (bacteria and viruses). To counter the sentinels of genome integrity, viral pathogens have evolved strategies to usurp these host replication and repair proteins for their own benefit. Viral strategies to hijack the nuclear compartment include: localizing to cellular sites of DNA damage, generating additional DNA breaks on the host, amplifying the cellular DNA damage signals and tethering themselves extra- chromosomally to DNA break sites to persist long-term. However, the mechanisms by which viruses carry out these pathogenic activities remain largely unknown. Better mechanistic understanding of how viruses dysregulate host DNA repair and replication pathways are essential for developing therapeutic interventions for human health. The goal of this research program is to understand how genome stability is impacted by viral pathogens by building on existing technologies to develop novel tools that will help us interrogate how replication and repair proteins are usurped, modified and leveraged by DNA viruses. We propose to answer fundamental questions about host genome stability that are utilized by viral pathogens for short-term and long-term benefit. We will determine how cellular signals are modulated on a global scale by DNA viruses and how these signals impact host replication proteins. We will then decipher how viruses utilize cellular signals to localize their proteins and genomes to cellular DNA break sites. Lastly, we will interrogate how cellular proteins are used by viruses to tether themselves in proximity to DNA break sites for long-term persistence. Not only are these fundamental discoveries, they also have the potential to inform cancer therapies and human DNA repair in addition to generating knowledge about viral life cycles. These basic studies on viruses and DNA damage will benefit a diverse array of biologists who study viral signaling in the biosphere. These strong foundational methodologies can be applied broadly to study all eukaryotic viruses as well as bacteriophages.