University Of Wisconsin-Madison

NIH Research Projects · FY 2025 · 2018-09

Project Summary The goal of this R01 renewal application is to further develop the LED-enhanced NMR technology (LC-photo- CIDNP) that was established during the prior cycle of funding, and extend the method to the facile and ultra- rapid 1D-to-3D NMR spectroscopy of proteins at sub-micromolar concentration. We will focus on NMR studies in solution and will target folded, unfolded and intrinsically disordered proteins in either buffered solution or cell- like media. We will accomplish the above goals within three steps. First (Specific Aim #1), we will incorporate a tryptophan (Trp) isotopolog bearing a quasi-isolated 1H-13C spin pair (QISP) within soluble proteins to achieve unprecedented NMR sensitivity for the detection of solvent-exposed Trp in proteins at nanomolar and sub-nanomolar levels. We will then employ the above technology in combination with field-cycling to achieve further NMR sensitivity enhancements. Second (Specific Aim #2) we will extend LC-photo-CIDNP to amino acids other than Trp and Tyr within proteins. This goal will be accomplished via through-space and through- bond polarization transfer methodologies. Third (Specific Aim #3), we will extend LC-photo-CIDNP to higher- dimensionality (>2D) NMR spectroscopy by developing novel 3D (and possibly 4D) 1H,13C heteronuclear spectroscopy pulse sequences tailored to the analysis of side-chain and backbone 1H-13C resonance pairs. This effort will include non-uniform-sampling (NUS) data collection schemes. We will then combine theoretical calculations and experiments to develop better LC-photo-CIDNP dyes with optimized g-factor values and long photoexcited-state lifetimes, for optimal LC-photo-CIDNP data collection. We will also exploit the peculiar field dependence of LED-enhanced NMR and implement 2D LC-photo-CIDNP on benchtop NMR spectrometers. Finally, we will test the success of the improved LC-photo-CIDNP technologies developed in this work by studying the interaction of an aggregation-prone client protein (SH3 variant) with the Hsp70 molecular chaperone at sub-micromolar concentration.

NIH Research Projects · FY 2025 · 2018-09

PROJECT SUMMARY/ABSTRACT Primates are uniquely capable of interpreting external stimuli and responding in behaviorally advantageous ways. A key neuronal process supporting these abilities is the sophisticated level to which their visual systems construct three-dimensional (3D) representations of the world from two-dimensional (2D) retinal images. Indeed, 3D spatial processing was a driving factor in the evolution of the primate brain and human analytical abilities. Today, deficits in 3D processing help define certain neurodevelopmental disorders. Our overarching hypothesis is that 3D visual perception, oculomotor processing, and the formation of sensorimotor associations that facilitate strategic behaviors are collectively supported by a little-studied V3A → caudal intraparietal (CIP) hierarchy that bridges occipital and parietal cortex. However, there is a critical gap in the understanding of these areas’ causal roles in perception and it is unknown if shared circuitry within this hierarchy jointly supports visual and oculomotor functions. Here we propose new experiments with macaque monkeys that combine behavioral, high-density electrophysiological, and causal manipulation techniques to fill these gaps. In Aim 1, we will assess the causal contributions of V3A and CIP to 3D perception. The experiments will use electrical microstimulation (EM) to manipulate neuronal activity in each area while the monkeys perform an eight-alternative forced-choice (8AFC) surface orientation (tilt) discrimination task. We hypothesize that weak currents applied to clusters of V3A/CIP neurons with similar visual selectivity will systematically bias the 3D orientations reported by the monkeys. The predictions for which neuronal/stimulus factors will determine the direction and magnitude of the induced biases are hypothesis driven and highly site specific. A comprehensive linear regression model will be used to test our hypotheses that biases will: (i) have distinct relationships with the stimulus tilt (relative to the preferred tilt at the EM site) and slant, (ii) be larger at EM sites with 3D object selectivity compared to lower-level visual feature selectivity, and (iii) be larger when CIP is stimulated than V3A. In Aim 2, we will test if the areas carry presaccadic activity and if training shapes sensorimotor associations between the neurons’ visual and saccadic properties. Specifically, in Aim 2A we will use overlap and memory-guided saccade tasks to test for presaccadic activity and evaluate two alternative hypotheses regarding sensorimotor associations in monkeys naïve to the 8AFC tilt discrimination task. Namely, whether an alignment of surface tilt and saccade direction preferences in V3A/CIP naturally occurs in the circuit or is flexibly learned and dependent on sensorimotor training. In Aim 2B, we will assess the mechanisms (visual and/or saccadic) supporting sensorimotor associative learning and their temporal dynamics by tracking the relationship between visual selectivity and presaccadic activity as a function of training (duration & performance level) on the 8AFC discrimination task. By testing the causal roles of these two interconnected cortical areas in 3D perception and if shared circuitry jointly supports visual/oculomotor functions, this work will advance the understanding of processes that enable primates to uniquely thrive in a 3D world.

NIH Research Projects · FY 2025 · 2018-07

Project Summary Cells live in diverse environments and cellular communities, from the cells in our bodies to single-celled organisms surviving in the soil. To navigate these complex environments, cells must be able to sense and respond to a variety of signals. This is done through biological signaling pathways, consisting of sensors and interacting proteins, which process external signals and transmit information. My research program focuses on understanding how these biological networks transmit information about external signals to the activity of intracellular effectors, such as transcription factors, to generate an appropriate cellular response or state and how these cell states affect community-level phenotypes. Understanding this signal processing represents a key gap in our knowledge of how healthy and diseased cells make decisions and guide the behavior of cellular communities. Specifically, we ask (1) How do signaling networks transform extracellular signals into appropriate intracellular signals? (2) How are intracellular signals interpreted by the cell to generate appropriate responses? and (3) How do individual cell decisions affect population-level community phenotypes? Our research is focused on understanding signaling specificity and kinetics in the mitogen-activated kinase (MAPK) pathways as well as transcription factor dynamics and subsequent gene expression in response to environmental stress. MAP kinase pathways are conserved from yeast to humans and control vital cellular processes including proliferation, differentiation, and stress response. We use a variety of systems to address the questions outlined in this research proposal including Saccharomyces cerevisiae, the human fugal pathogen Candida albicans, synthetic signaling pathways, and mammalian cell culture. We take a multi-pronged approach that uses microfluidic and optogenetic tools to perturb signaling pathways and combine these perturbations with mathematical modeling to understand how different properties of signaling pathways, including bandwidth and crosstalk, allow them to appropriately transform their input signals. Furthermore, we use these tools to drive dynamics of intracellular effectors, such as transcription factors, and ask how these different effector dynamics generate cellular responses. And finally, we use the exquisite spatiotemporal control available with light to generate desired states in individual or populations of cells, including fungal biofilms, and ask how this affects community-level phenotypes.

NIH Research Projects · FY 2025 · 2018-07

Abstract In 2007, some pets became ill and a few died as the result of consuming contaminated pet food. An investigation revealed that the incident was due to melamine, an adulterant found in the contaminated pet food. Melamine was also found in tainted animal feed that was used for farm animal and fish. Some food animals that ate the tainted feed were processed into human food. This event had major implications for animal and human health. In recognition of the event and its consequences, the Center for Veterinary Medicine (CVM) sought out cooperative agreements with veterinary diagnostic laboratories to enable analysis of animal diagnostic samples and animal food/drug products in the event that laboratory surge capacity resulting from large-scale outbreak or threat incident is needed. Participating laboratories have increase surge capacity and have prepared for analysis related to microbiological or chemical contamination, either through intentional or unintentional means. This consortium of laboratories is useful for the detection and surveillance of animal feeds or other large-scale outbreaks, so as to halt an event early and reduce consequences. The emergent of SARS-CoV-2 virus in 2019 was a public health need that was not identified at the time of our original Vet-LIRN award. The susceptibility of multiple animal species including household pets to the virus generated an unknown threat to public health, and warrant investigation to bridge the knowledge gap of the virus at the human-pet/animal interface. This application is to continue the Wisconsin Veterinary Diagnostic Laboratory’s (WVDL) commitment to this cooperative agreement and to establish rapid communication with Vet-LIRN to increase the government’s ability to examine samples from animals adversely affected by contaminated or adulterated products. Examination of such samples can contribute to overall food safety as animal food events could signal potential issues in the human food system. The University of Wisconsin (UW)- Madison, Wisconsin Veterinary Diagnostic Laboratory (WVDL) is fully accredited by the American Association of Veterinary Laboratory Diagnosticians (AAVLD), and thus, has the personnel with necessary experience, technical expertise and necessary infrastructure to participate in method standardization, training, proficiency testing, and deployment of new equipment to accomplish the task described above with relative ease. The WVDL has participated in several proficiency test and intra- laboratory comparison exercises (ICE) in the past 5 years and has participated in other funding opportunities included the purchase, implementation and utilization of the Illumina iSeq 100 sequencing platform and SARS-CoV-2 PCR and serology testing.

NIH Research Projects · FY 2025 · 2018-05

ABSTRACT The primary focus of my laboratory is the development of new tools and strategies for proteomic analyses of complex biological systems, specifically centered around the concept of the proteoform. Proteoforms, each of which comprises a unique combination of amino acid sequence and post-translational modifications (PTMs), are the primary molecular effectors of cell function. Subtle sequence and PTM differences between proteoforms can completely alter their function and activity. We see comprehensive proteoform-level analysis of biological systems as absolutely essential to understanding their function, for both individual pathways and networks operative within cells, and more globally, to decipher the systems-biology-level dynamics and interactions that control cellular response. The current technology for global proteoform analysis in complex systems is in its infancy, offering both a great challenge and a great opportunity. Our laboratory is keenly interested in tackling this problem and is pioneering a new approach that integrates high resolution proteoform intact mass measurements, both bottom-up and top-down strategies, new informatic tools for the comprehensive analysis of PTMs, and RNA-Seq information; all woven together in a robust bioinformatic framework to allow the comprehensive identification and quantification of proteoforms in complex mixtures. Along with other world-class scientists, we will work towards embarking on the Human Proteoform Project, which includes ambitious subprojects describing the construction and utility of comprehensive proteoform atlases for humans and model organisms. Specific projects in our laboratory will include development of the following: (1) a multi-dimensional separation strategy for increased breadth and depth of proteoform identifications; (2) a source-induced dissociation method for fragmentation of eluting proteoform ions to increase proteoform identifications; (3) intelligent real-time data acquisition; (4) direct acquisition of orbitrap time-domain transients to expand the accessible mass range; (5) data analysis software including the abilities to search for truncated proteoforms and utilize the most abundant mass for identification; (6) sample-specific databases created through integration of bottom-up, top-down, intact mass and RNA-Seq data; (7) visualization tools for manual validation of proteoform identifications and for troubleshooting problems with samples and/or algorithms; and (8) using proteoform quantitative trait loci (QTLs) to reveal the modifying enzymes encoded elsewhere in the genome that are responsible for the critical post-translational modifications with functional consequence. We are excited to develop powerful new tools to advance the state-of-the-art in this new and important field of study to reveal the biologically important effectors of cellular mechanisms. These tools, which will be made widely available to all researchers, will reveal new information essential to the understanding of both normal and disease biology, deepening and accelerating the study of human disease processes.

NIH Research Projects · FY 2026 · 2018-05

Project Summary Bacteria frequently encounter stresses including nutrient starvation, temperature changes, and antibiotic assault, which could easily throw their intracellular environment into chaos. To survive and to adapt, bacteria developed diverse stress responses to regulate intracellular processes accordingly. While the transcriptional networks governing stress responses have been extensively characterized, there are major gaps in our knowledge beyond transcription regulation. The theme of my research is to elucidate stress signaling mechanisms that are transmitted by rapid changes in concentration of ‘alarmones’ – signaling nucleotides which are instrumental for alerting cells about stresses in a timely manner. My laboratory has extensive experience in characterizing the conserved alarmone (p)ppGpp. (p)ppGpp is induced by stresses and mediates profound, pleiotropic physiological changes in almost all bacteria to allow fitness, survival, and evolution. We identified multiple purine synthesis enzymes, a replication enzyme and a transcription repressor that are directly regulated by (p)ppGpp in Gram-positive Bacillus species. These regulations were further found to be conserved in many pathogens and are critical for homeostasis, starvation resistance, antibiotic persistence, and genome integrity. Currently, we are also investigating how (p)ppGpp regulates the switch between distinct bacterial lifestyles: planktonic growth and biofilm formation. Additionally, we detected other nucleotide alarmones including AppppA, pGpp, ppApp, and c-di- AMP, which are induced by different stresses including temperature and cell wall stress, to form a robust protective network. Our future research will answer the following fundamental questions: How are the different alarmones triggered by different stresses, and how do bacteria synthesize them? What are the direct interaction targets of different alarmones, and how do they promote bacterial fitness and influence bacterial development such as biofilm formation and sporulation? How do bacteria integrate multiple cues from different alarmones for rapid and appropriate adaptation to diverse environments? We combine metabolomics, transcriptomics, and proteomics with biochemical and cell biological approaches to answer these questions. We obtained a list of alarmone targets from systematic screens performed with the proteome of the pathogen Bacillus anthracis. We will study these processes in the related non-pathogenic bacterium Bacillus subtilis for which we have extensive experience. B. subtilis grows fast and is highly amenable to genetic manipulation. The nucleotide signaling mechanisms we characterize in Bacillus are applicable to other, less tractable, pathogenic bacteria, and can be used for developing antimicrobial strategies by targeting their stress responses.

NIH Research Projects · FY 2026 · 2018-04

Abstract HCMV encodes multiple proteins that suppress lytic phase immediate early (IE) transcription during latency. These include Us28 and UL138. We identified a new repressor of the MIEP during latency: the Pentamer. The Pentamer is a five-member glycoprotein complex consisting of gH, gL, UL128, UL130, and UL131 that is required for entry into epithelial and endothelial cells. We determined the Pentamer represses the MIEP during both lytic and latent infections. Provocatively, we determined at least two different Pentamer alleles exist, one that strongly represses the MIEP, and one that weakly represses the MIEP. We hypothesize that HCMV uses unique Pentamer alleles to create different sub-sets of virions pre-programmed to either initiate a lytic infection or to establish latency. To test this hypothesis, in Aim 1 we will create genetically matched virus strains (laboratory and clinical) with pro-lytic or pro-latency Pentamer alleles and test them for their propensity to initiate lytic infection in fibroblasts, epithelial and endothelial cells, to select against Pentamer function during fibroblast propagation, and to establish latency in primary CD34+ cells. In Aim 2 we will determine the mechanism through which the Pentamer represses the MIEP.

NIH Research Projects · FY 2025 · 2018-03

Project Summary/Abstract Our group completed the natural history study of premanifest Huntington disease (HD) entitled Predict-HD which evaluated over 1400 research volunteers who were healthy but had undergone the predictive test for the gene that causes HD. Findings revealed that signs and symptoms of HD were evident up to 15 years before the traditional diagnosis of HD was given in the clinic. From these data we were able to develop models of prognosis, disease progression and prediction of HD onset. Disease-modifying clinical trials are currently underway to slow the progression, or delay the onset, of HD. More recently, a collaborative group published an assay to measure the amount of mutant huntingtin protein in the cerebral spinal fluid of HD participants. Questions of central importance to the success of this measure for clinical trials require investigation: (1) how reliable is the measure in the same person when repeated (intra-subject test-retest reliability); (2) how reliable is the measure in the same person when analyzed by two different labs (inter-lab reliability); (3) does the measure reflect disease symptoms (content validity); (4) does the measure predict meaningful disease outcomes (prognostic validity); (5) does the measure track disease progression or severity; and (6) how many (and what stage of HD) research subjects do we need to know with confidence that an intervention is working (i.e., delaying onset/slowing progression)? The proposed research study will address all of these limitations to more effectively test new experimental interventions such as gene therapies and new drugs. Findings will immediately inform how the field should best design preventive clinical trials for HD.

NIH Research Projects · FY 2026 · 2018-03

Project Summary The prefrontal cortex (PFC) supports a constellation of ‘executive’ cognitive processes that guide goal-directed behavior. Dysregulation of PFC-dependent cognition is associated with numerous behavioral disorders. Currently there is a strong need for improved treatments for PFC- dependent cognitive dysfunction. However, the development of novel treatments is hindered by our limited understanding of the neurobiology underlying PFC-dependent cognition. In recent studies we demonstrated that corticotropin-releasing factor (CRF) neurons in the caudal, but not rostral, dorsomedial PFC (dmPFC) of male and female rats (outside proestrus) impair two distinct cognitive processes: working memory and sustained attention. Conversely, inhibition of PFC CRF neuronal activity or blockade of CRF receptors, locally or globally in the brain, improved PFC- dependent cognition. Interestingly, the regulatory actions of CRF across these distinct cognitive processes involve distinct pathways: local release for working memory and extra-PFC release for sustained attention. The mediodorsal nucleus of the thalamus (MDthal) plays a central role in the regulation of PFC-dependent function. Preliminary studies indicate that MDthal plays a prominent role in the sustained attention actions of PFC CRF neurons. We recently demonstrated that the PFC CRF neurons are comprised of both glutamatergic (CRFGlu, 85%) and GABAergic (CRFGABA, 15%) subpopulations. The proposed multidisciplinary studies will provide a better understanding of neural mechanisms that underlie the cognitive actions of PFC CRF neurons. Aim 1 uses recently developed viral vector-based chemogenetic manipulations to determine the cognitive actions of CRFGlu and CRFGABA neurons. Aim 2, building on preliminary observations tests the hypothesis that the sustained attention (an possibly working memory) actions of PFC CRF neurons involve projections to the MDthal. Aim 3 examines the neural coding actions of caudal dmPFC CRFGlu and CRFGABA across the PFC-MDthal circuit. Collectively, these studies will provide novel insight into the neurobiology of PFC-dependent cognition. In doing so, these studies may provide a better understanding of the neural bases of PFC cognitive dysfunction and lead to novel treatment strategies for PFC-dependent cognitive dysfunction.

NIH Research Projects · FY 2026 · 2018-03

ABSTRACT Mechanical load is a fundamental regulator of cardiac function. The heart operates in a dynamically changing mechanical environment, and alterations in intra-cardiac pressure and/or volume preload/afterload influence cardiac performance to coordinate cardiac output with venous return and arterial blood supply. A crucial aspect of this regulation is the modulation of heart rate, which is controlled by the sinoatrial node (SAN), the primary pacemaker of the heart. The SAN anatomy and location within the heart enable it to detect fluctuations in both coronary and atrial blood pressure, establishing a structural foundation for the regulation of heart rate through SAN mechanosensitivity in response to hemodynamic changes. Although physiological stretch is an essential component of the SAN autoregulatory feedback mechanism, chronically elevated stretch results in severe myocardial remodeling and leads to SAN dysfunction (SND), also known as sick sinus syndrome. Conditions associated with mechanical overload, such as hypertension, often exhibit SND, which manifests as bradycardia, irregular atrial pauses, and sinus arrest/block. The upstream mechanisms of SND in the hypertensive heart are unexplored and contribute to lack of preventative intervention. We will address this gap in knowledge by employing a combination of several multi-level cutting-edge imaging modalities of cellular microarchitecture, Ca2+ and cAMP dynamics, electrophysiological measurements, biochemical studies, and computational modeling to demonstrate an innovative concept that proposes a tight association between mechanical loading, SAN pacemaking, and its regulation by the autonomic nervous system through a mechanosensitive caveolar pacemaker signalosome. This caveolar domain provides the spatiotemporal foundation for mechano- electrochemical signal transduction and heart rhythm regulation, which involves stretch-induced augmentation of cAMP production and cAMP/PKA-mediated phosphorylation of Ca2+ handling and sarcolemmal proteins as well as activation of caveolar mechano-sensitive ion channels. We propose that prolonged (chronic) atrial overload leads to the degradation of caveolae, causing SND and an altered response to both mechanical and autonomic stimulation, which forms the molecular basis for chronotropic incompetence. Preventing caveolae degradation or restoring caveolae structures could alleviate SND phenotype and improve the SAN ability to adequately respond to emotional or physical stressors. This research holds significant potential impact as it will provide mechanistic insights that can serve as a foundation for developing innovative therapeutic strategies aimed at preventing SND in hypertensive individuals.

NIH Research Projects · FY 2026 · 2018-01

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease (AD) is the most common form of dementia in the elderly population and 6th leading cause of death in the US. Despite extensive research, there are currently no treatments that slow or stop the development of AD. With the number of AD cases expected to triple in the next 30 years, there is a pressing need to diagnose AD early in the preclinical stage. While several peptide and protein biomarkers in cerebrospinal fluid (CSF) have been used for AD diagnosis, an unequivocal diagnosis in the early phases of AD is still lacking. Perhaps more importantly, the discovery and establishment of reliable biomarkers capable of monitoring progression and degree of cognitive impairment as well as potential efficacy of therapy remains a major challenge. Furthermore, compared to CSF, serum sample provides an appealing source for biomarker discovery and screening due to less invasiveness and easier access. However, the correlation between CSF and blood protein/peptide biomarkers as well as changes in the brain structure/function and cognition in AD is not well established. In order to address these challenges and fill in existing knowledge gaps, we propose to employ a multi-faceted approach combining a suite of mass spectrometry-based technologies enabled by innovative multiplexed tagging strategies, improved sampling and separation strategies and clinically-available measures to discover, identify and evaluate candidate biomarkers of AD in CSF/serum obtained from asymptomatic cognitively-healthy middle-aged adults, older cognitively-normal adults, and patients with mild cognitive impairment (MCI) and AD. We propose the following specific aims: Specific Aim 1 – To develop novel enrichment strategies and complementary separation modalities for enhanced coverage of glycoproteome and posttranslational modification crosstalk analysis in paired CSF and serum samples from subjects in control, preclinical, MCI, AD groups, respectively. Specific Aim 2 – To enhance quantitative glycoproteomic analysis of low-abundance species in CSF and serum samples via innovative dimethylated leucine (DiLeu) boosting and BoxCar data-independent acquisition (DIA) strategies along with machine learning classification algorithms for improved diagnosis of AD. Specific Aim 3 – To validate candidate AD biomarkers, in CSF and serum samples collected from individuals with MCI and dementia, using targeted quantitative proteomics approaches enabled by isotopic DiLeu tags and affinity-bead assisted MS immunoassay along with association with AD-related clinical, cognitive and neuroimaging measures. This project uniquely integrates advances in MS-based multiplexed quantitative glycoproteomics and bioinformatics tools with neuroimaging and clinical measures to enable more comprehensive discovery and validation of CSF and serum biomarkers in AD. These biomarkers would be invaluable in improving our understanding of AD pathogenesis, designing therapeutics for patient care and more efficient clinical trials of disease modifying therapies. The advances in technology and new insights will have broad impact on translational medicine.

NIH Research Projects · FY 2026 · 2018-01

Project Summary Age-related diseases are the major causes of morbidity and mortality in the US. Many elderly people suffer from multiple age-related diseases simultaneously; while the risk of almost every individual disease rises with age, they also interact. For example, diabetes and obesity are risk factors for neurodegenerative diseases including Alzheimer’s disease (AD). Calorie restriction (CR), a dietary intervention which extends lifespan while delaying or preventing age-related disease, is one plausible approach to lessen the burden of multiple age- related diseases simultaneously, but reduced-calorie diets are notoriously difficult to sustain. Recent studies have highlighted an important role for dietary protein in health and longevity, with protein restriction (PR) shown to promote longevity and to mimic the metabolic, frailty, and cognitive benefits of CR. During the initial project period, we found that specifically reducing dietary consumption of the three branched-chain amino acids (BCAAs) – leucine, isoleucine, and valine – has sex-specific benefits for frailty and lifespan in C57BL/6J mice. We determined that the metabolic and molecular effects of PR are both sex and strain dependent, and that the role of a specific hormone proposed to mediate the effects of PR may be more limited than previously suspected and also differ between sexes and strains. Finally, we found that the BCAAs have distinct roles on metabolism, with restriction of isoleucine being necessary and sufficient for the metabolic benefits of PR. In preliminary experiments, we have also found that isoleucine restriction has sexually dimorphic effects on healthspan and longevity in genetically heterogenous mice, and that PR has beneficial effects on cognition and disease pathology in a mouse model of Alzheimer’s disease. Here, we will rigorously test the ability of graded restriction of isoleucine to promote health and longevity in DBA/2J and C57BL/6J mice of both sexes, examining the effects on metabolic health, frailty, cognition and lifespan as well as the effects on pathology and at the molecular level. We will identify the role of a specific hormone, FGF21, in the metabolic response to isoleucine restriction. Finally, we will test if restriction of individual BCAAs is necessary and sufficient for the ability of a PR diet to prevent or delay AD. The proposed work will examine the role of the BCAA isoleucine on health and longevity in multiple genetic backgrounds for the first time and answer long-standing questions regarding how dietary composition impacts healthy aging. Importantly, we will gain new insight into the mechanisms that drive the potent effects of isoleucine restriction on healthy aging, and break new ground identifying how individual BCAAs impact the progression of AD. In the long term, this work will enable our lab and others to develop a mechanistic understanding of how dietary BCAAs and other macronutrients regulate health and disease vulnerability, and to identify new targets for pharmacological treatments to promote healthy aging.

NIH Research Projects · FY 2025 · 2017-09

PROJECT SUMMARY/ABSTRACT Recipients of a kidney transplant from an African-American deceased donor have worse outcomes than their counterparts receiving an organ from a European-American deceased donor. In addition, kidney transplants from deceased donors with two apolipoprotein L1 gene (APOL1) high-risk genotypes, which is almost exclusively found in individuals of recent African ancestry, have shorter survival. Some recipients of these organs, however, have good long-term outcomes. Therefore, we hypothesize that APOL1 genes interact with other environmental or inherited factors to cause accelerated failure of kidney transplants. Similarly, living donors of recent African ancestry face increased risk of post-donation kidney failure compared to European-American living donors, but the potential impact of APOL1 genotypes in these donors is unclear. The National Institutes of Health (NIH) established the “APOL1 Long-term Kidney Transplantation Outcomes” (APOLLO) U01 Consortium in 2017 to prospectively address several questions regarding APOL1 genotyping in kidney transplantation. The APOLLO Consortium includes a Scientific and Data Research Center and 13 Clinical Centers, including our University of Wisconsin Clinical Center. In the proposed APOLLO Phase 2, we will collect extended long-term data on graft function, injury, and survival to more comprehensively define outcomes associated with donor and recipient APOL1 genetic variants and continue to recruit new living donors of African ancestry. Working with the SDRC, we will return APOL1 genotype results to enrolled transplant recipients and living donors. The data to be collected will define the impact of genotypes on graft outcomes and living donors and identify secondary factors that modify outcomes. These data are critical to improve allograft survival and quality of life for all transplant recipients.

NIH Research Projects · FY 2025 · 2017-09

The central goal of the Institute for Clinical and Translational Research (ICTR) TL1 Training Program is to contribute to translational workforce development by preparing a robust cohort of scholars that can address societal health needs through clinical and translational research (CTR) endeavors in academia, government, and industry. The University of Wisconsin-Madison (UW) TL1 Training Program has been active since 2008, and currently supports both predoctoral and postdoctoral scholars for two-year appointments. The objectives for the upcoming funding period are to: 1) Augment our current success in building capacity of the translational workforce via predoctoral and postdoctoral training; 2) Innovate our tailored, competency-based TL1 curriculum to provide the flexibility and adaptability to realize the “Wisconsin Idea” in the context of a continually shifting research landscape; 3) Advance a positive environment by intentionally engaging a broadly representative translational workforce and expanding the research portfolio to pursue health for all people; and 4) Instill trainees with the team science and leadership skills to work effectively in interdisciplinary teams. We will address these objectives by providing more flexible curricular options in the areas of community outreach, bioinformatics, science communication, and entrepreneurship; by enhancing experiential learning opportunities through biotechnology externships in partnership with UW’s Forward BIO Innovators in Training program; and by promoting inter-hub collaboration and team science initiatives by instituting an annual Midwest TL1 Research Summit. We are requesting support for 11 predoctoral and four postdoctoral scholars in the upcoming period. All scholars will receive didactic training in core translational research skills, the responsible conduct of research, rigor and reproducibility, mentor/mentee strategies, and team science skills. We will evaluate the success of our program guided by the ICTR Innovation Scorecard and measure impact using the Translational Sciences Benefits Model. We will use mixed qualitative and quantitative methods, including monitoring scholar progress along individual development plans and self-efficacy in key National Center for Advancing Translational Sciences CTR competencies; tracking scholar productivity and career milestones; surveying scholar satisfaction with training opportunities; and continually assessing our scholar cohorts and project portfolios to advance a broad representative workforce. This innovative TL1 programming, including mentorship, team science, a focus on health for all, and a positive environment, will foster and launch a cadre of independently funded leaders of CTR who will impact the health of our patients, families, and communities.

NIH Research Projects · FY 2025 · 2017-09

The University of Wisconsin (UW) Institute for Clinical and Translational Research (ICTR) KL2 Program, launched in 2007, has recruited and trained 82 scholars varied in their discipline, area of study, and position along the translational spectrum. We appoint three scholars per year for four years using a combination of internal and external funds. Graduated scholars remain committed to research (90%) in academics or industry, have been promoted or are eligible for promotion (87%), have successfully competed for funding as a principal investigator/multiple principal investigator ($225 million) and as a co-investigator ($288 million), and have published prolifically as first/senior author (513) and as a co-author (1,456). To accelerate these and other successes, ICTR has completed an institute-wide strategic plan (Innovation Scorecard) that serves as a roadmap to use continuous improvement to address ongoing challenges in conducting efficient, rigorous, and engaged CTR at our Hub—with specific attention to CTR capacity building. In concert with these continuous program improvement efforts, we engaged a comprehensive KL2 needs assessment, which recognized key challenges to early-career scientists, including: 1) maintaining work-life integration, 2) transitioning to independence, 3) promoting broadly representative research teams and study participants, 4) designing research for dissemination, and 5) disseminating and implementing research to improve health outcomes. In the next grant period, we propose to refine our KL2 Program to address these challenges by achieving the following Specific Aims: 1) Enhance scholar’s vitality by building their capacity to adjust to life and career stressors, 2) Foster scholars’ engagement of community members in research conduct and dissemination, 3) Establish longitudinal programming to help scholars incorporate dissemination and implementation concepts into their research programs, and 4) Disseminate the KL2 Program’s mentorship and coaching innovations. We have developed a comprehensive plan to increase the recruitment and retention of broadly representative scholars.. We have also designed a rigorous program evaluation plan to assess influence at all levels of the translational research ecosystem. Program success will be measured at each level: 1) individual: secure grants and publish work in high-impact journals; 2) proximal: develop and maintain relationships with mentors, mentees, and research teams; 3) institutional and inter-institutional: form inter-disciplinary scientific teams and transform clinical care and UW culture; and 4) societal: revolutionize clinical guidelines and improve health for all.

NIH Research Projects · FY 2025 · 2017-09

The University of Wisconsin-Madison (UW) Institute for Clinical and Translational Research (ICTR) collaborates with partnering schools and the Marshfield Clinic Research Institute to expand support and innovation to accelerate clinical and translational research (CTR) across our Hub. Over the past funding period, our KL2 scholars have successfully competed for $131M in extramural funding. Our evidence-based CTR mentorship trained 501 master facilitators in 51 CTSAs. We engaged regional networks in 90% of the counties in Wisconsin. Recently, ICTR led multiple impactful mid-course corrections enabling our students, staff, and scientists to effectively address health challenges revealed by the COVID-19 pandemic. Building on user needs assessments, SWOT analyses, and institute-wide strategic planning we developed an Innovation Scorecard that serves as a roadmap to use continuous improvement to address ongoing challenges in conducting efficient, rigorous, and engaged CTR. Our application addresses 5 impactful Aims aligned with objectives in our Innovation Scorecard, each testing explicit hypotheses. These Aims are: 1) Expand skills, efficacy, and resiliency of the CTR workforce to engage effectively in multidisciplinary approaches to CTR. We test an eco-systems approach to promote trainee resiliency, and propose to enhance academic recognition of clinical research, mentorship, and team science. 2) Use bidirectional population engagement and recruitment approaches to increase representative CTR leading to improved health for all in Wisconsin. We will test whether incorporating innovative stakeholder engagement, dissemination and implementation, and team science approaches improve the fidelity and sustainability of interventions in health systems and communities. 3) Employ intentional, systems-level change strategies to advance a positive environment for all at UW. We test evidence-based interventions to engage a broad representative workforce and support translational teams to engage broad populations in CTR. 4) Support a sustainable informatics ecosystem to transform data to knowledge, enhance integration of research into health care delivery, and promote rigor and reproducibility in CTR. We will provide and support secure data sharing, interoperable systems, disease-focused registries and well-trained informaticians to advance data-driven CTR. 5) Integrate a “concept-to-closure” support model for clinical research engaging UW Health regional and national networks. We integrate support for CTR within UW Health. ICTR’s broad reach, experienced leadership, high-functioning infrastructure, and partnerships will enable us to advance translational science as a rigorous discipline, providing disruptive innovations in CTR.

NIH Research Projects · FY 2025 · 2017-08

Abstract The essential human enzyme O-GlcNAc transferase (OGT) catalyzes a unique type of intracellular protein glycosylation called O-GlcNAcylation. In response to nutrient levels and stress, OGT dynamically regulates a variety of physiological and pathological processes including the “Warburg effect” in cancer cells and insulin resistance in diabetes. Previous studies on the OGT active site have made fundamental discoveries on its catalytic mechanism and substrate interactions. However, how OGT regulates protein- and site-specific O- GlcNAcylation remains unclear. This is due to a number of challenges including: 1) OGT glycosylates thousands of proteins without a conserved sequence motif near the O-GlcNAc modification site, 2) a majority of O- GlcNAcylation sites are found on intrinsically disordered regions (IDRs), 3) OGT typically binds proteins with low/moderate affinity, and 4) a lack of OGT-protein complex structures. In our last funding period, we have made strides in these areas through development of a suite of novel chemical probes that allow us to interrogate OGT specific interactions with low/moderate affinity for structural, proteomic, and biochemical characterizations. This proposal aims to make further conceptual and technical breakthroughs toward addressing these longstanding challenges. It is expected that a better understanding of how OGT interacts with other proteins, particularly through the regions beyond the OGT catalytic site, will be essential for understanding OGT’s functional regulation at protein- and site-specific levels, filling major knowledge gaps between decades of biological observations of OGT’s nutrient sensing and other regulatory roles, and will support the need to specifically modulate OGT functions for biomedical applications.

NIH Research Projects · FY 2025 · 2017-08

PROJECT SUMMARY/ABSTRACT Human Papillomaviruses (HPV) cause 5% of human cancers. While there are effective prophylactic HPV vaccines, their poor uptake in the US, their inaccessibility in many parts of the world where these cancers are most frequent, and the fact they do not eliminate pre-existing persistent infections that can lead to cancer, and they do not treat the resulting cancers, requires that we continue advancing studies on these important human tumor viruses. During the current, highly productive funding period for this R35 (58 publications) we made many, important, new insights that shed light on how papillomaviruses (PV) i) evade host immunity to establish persistent infections that lead to cancer, ii) how this same mechanism contributes to resistance to immunotherapy in mice and humans, iii) how estrogen alters systemic and host immunity to drive persistent PV infections and PV-induced disease, iv) that PV-induced cancers arise from epithelial progenitor cells, v) that PVs alter the microbiome, and vi) the role of host genes in PV-associated cancers. Many of these studies arose through our broad study of a recently discovered mouse papillomavirus (MmuPV1) that we and others have demonstrated induces the same range of cancers caused by HPVs in humans, is sexually transmitted as with HPVs that cause cancer, and shows a similar propensity and mechanism of integration into the host genome, as seen with HPVs, while other studies made use of our first and new, second-generation HPV transgenic mouse models. We also made important new gains to our knowledge of Merkel cell polyomavirus and its role in human cancer, and we contributed to numerous collaborative studies with other leading labs in the field of tumor virology. The competitive renewal application of this R35 builds extensively on these important gains in knowledge. Broadly, our proposed studies are focused on three general directions of study: 1) understanding the role of host immunity in papillomaviral pathogenesis; 2) characterizing MmuPV1-induced cancers and their relevance to HPV-related cancer; and 3) defining the interplay between the microbiome and papillomavirus-induced disease. The R35 mechanism has allowed my research program to pursue many new avenues of research that have yielded significant, field-altering discoveries. We will apply and build upon these discoveries in our future research directions of this competitive renewal. We will leverage our expertise in animal model development and innovative, state-of-the-art approaches to pursue experiments aimed at answering many of the new questions raised by our studies over the current funding period. In the process, I will continue to train the next generation of scientists, as demonstrated by the fact that three trainees during the current funding period took faculty positions at highly ranked universities across the US. I am excited to be nominated by the Dean of the University of Wisconsin School of Medicine and Public Health to submit this competitive renewal application.

NIH Research Projects · FY 2025 · 2016-09

Asthma affects approximately 10% of US children and is a leading cause of respiratory morbidity and hospitalization. The large population of the Environmental Influences on Child Health Outcomes (ECHO) study that includes children from all over the USA is ideal for identifying early-life causes for childhood asthma. The Wisconsin Infant Study Cohort (WISC) is a birth cohort of rural families and children, including those living in small towns and on farms. This population would bring unique data to ECHO related to home and neighborhood exposures (e.g., animals, microbiome, green space), and health outcomes (e.g., reduced rates of respiratory diseases). Our scientific goals focus on how environmental factors and hormonal influences in adolescence regulate molecular responses of nasal airway cells (NAC). We will analyze DNA methylation (DNAm) and gene expression from NAC samples in mid-childhood and early adolescence and combine these data to identify “molecular phenotypes.” We hypothesize that these phenotypes relate to specific environmental exposures in early life and asthma-related outcomes at ages 6-10 and during a three-year follow-up period. We therefore propose the following specific aims: Aim 1. To leverage ECHO Protocol 3.0 core data, we will analyze nasal cell gene expression and DNAm in children ages 6-10 years to identify airway cell molecular phenotypes and then test for associations with prenatal and early postnatal environmental exposures, personal factors, and clinical outcomes (asthma, rhinitis, lung function). Aim 2. We will reassess asthma outcomes and NAC three years later (ages 9-13 years) to determine how asthma disease activity and changes in severity relate to: a) the molecular phenotypes at 6-10 years, b) potential asthma modifying factors such as puberty, insulin resistance, and allergy, and c) changes in DNAm and gene expression. Aim 3. We propose updating and adapting existing WISC protocols, adopting new ECHO systems, and implementing the ECHO Cohort Protocol with high fidelity to maximize retention of existing participants, contribute to rural and farming exposures and lifestyles, and enhance the ECHO Cohort Protocol. These proposed studies will link modifiable environmental exposures to molecular regulation of airway cells and allergy and asthma clinical outcomes. The results will yield a treasure trove of information that could inform new strategies to prevent asthma and, in affected children, enable innovative approaches to promote disease control and remission.

NIH Research Projects · FY 2024 · 2016-09

PROJECT SUMMARY Asthma affects approximately 10% of US children and is a leading cause of respiratory morbidity and hospitalization. Asthma disproportionally affects parent-identified Black and Caribbean Hispanic children, and the large and diverse population of the Environmental Influences on Child Health Outcomes (ECHO) study is ideal for identifying early-life causes for asthma disparities. The Wisconsin Infant Study Cohort (WISC) is a birth cohort of underserved rural families and children. This population would bring unique data to ECHO related to exposures (animals, microbiome), neighborhood factors (low population density, clean air), and health outcomes (reduced rates of respiratory diseases). Our scientific goals focus on how environmental factors and hormonal influences in adolescence regulate molecular responses of nasal airway cells (NAC). We will analyze DNA methylation (DNAm) and gene expression from NAC samples in mid-childhood and early adolescence and combine these data to identify “molecular phenotypes.” We hypothesize that these phenotypes relate to specific environmental exposures in early life and asthma-related outcomes at ages 6-10 and during a three year follow- up period. We therefore propose the following specific aims: Aim 1. To leverage ECHO Protocol 3.0 core data, we will analyze NAC gene expression and DNAm in children ages 6-10 years to identify airway cell molecular phenotypes and then test for associations with prenatal and early postnatal environmental exposures, personal factors (sex, parent-identified race/ethnicity, age, polymorphisms of candidate genes), and clinical outcomes (asthma, rhinitis, lung function). Aim 2. We will reassess asthma outcomes and nasal airway cells three years later (ages 9-13 years) to determine how asthma disease activity and changes in severity relate to: a) the molecular phenotypes at 6-10 years, b) potential asthma modifying factors such as sex hormones, insulin resistance, and allergy, and c) changes in DNAm and gene expression. Aim 3. We propose to update and adapt existing WISC protocols and adopt new ECHO systems to maximize retention of existing participants, contribute diversity related to rural and farming exposures and lifestyles, and implement the ECHO Cohort Protocol with high fidelity. These proposed studies will link modifiable environmental exposures to molecular regulation of airway cells and allergy and asthma clinical outcomes. The results will yield a treasure trove of information that could inform new strategies to prevent asthma and, in affected children, enable innovative approaches to promote disease control and remission.

NIH Research Projects · FY 2025 · 2016-08

PROJECT SUMMARY/ ABSTRACT. There continues to be a fundamental gap in understanding how CRISPR- based genome editors produce gene modifications in different human cells. A lack of understanding of why various editors fail and why some succeed in creating desired gene edits - while retaining full cell and tissue functionality - limits the use of genome editing tools. By observing genome editing in real-time within patient- derived cells in vitro, I seek to understand the bottlenecks in performing genome editing on human cells with precisely-controlled genome editor particles. Particles will be systematically assembled with various DNA, RNA, and polymeric components and delivered to patient-derived cells and microtissues. In situ high content imaging and analysis within customized cell substrates will monitor genome editing at multiple scales. The central hypothesis is that new assemblies of CRISPR-Cas9 particles can probe different biological processes of trafficking, DNA-double strand break formation, and DNA repair involved in the genome editing of human cells and tissues, as well as downstream effects on biological processes involving cell cycle arrest and morphogenesis. This hypothesis will be tested within patient-derived stem cells and tissues for both gene disruption and correction. An overarching rationale for the proposed research is that an improved understanding of fundamental biological processes involved with genome editing could enable the development of novel cell therapies and gene therapies for future genomic and precision medicine. Guided by strong productivity in the current early stage R35 award, I will pursue three research programs: 1) Assemble Cas9 particles to identify chromatin structures within human cells that promote gene correction; 2) Assemble Cas9 particles to identify delivery and DNA repair processes that promote gene correction within stem cells; and, 3) Assemble Cas9 particles to identify cell proliferative and tissue morphogenesis processes that promote gene correction of diseased mutations in patient-derived microtissues. Under the first research program, editing will occur at target genes that have variable chromatin structures within induced pluripotent stem cells (iPSCs), differentiated progeny, and with small-molecule treatment. Under the second and third research programs, genome editors will be applied to gene-correct diseased mutations in iPSCs, and microtissues matured from them. The approach is innovative, in the applicant’s opinion, because it departs from the status quo by systematically changing multiple components at a time using novel methods in patient-derived cells. The proposed research is significant because it is expected to advance and expand our understanding of how genome editing tools can be applied for the generation of advanced therapeutics, ranging from targeted small molecules to cell/tissue therapies. Ultimately, such knowledge would solidify the foundation for new translational projects involving genome editing.

NIH Research Projects · FY 2025 · 2016-08

PROJECT SUMMARY A longitudinal multi-omics examination of beta amyloid deposition (A), pathologic tau (T), neurodegeneration (N), and cognitive decline in the years prior to Alzheimer’s disease (AD) diagnosis is critical to better understand, predict, prevent, diagnose, and treat the disease. Gaps in knowledge include the timing, trajectory, and etiology of metabolite changes in the disease process. This renewal application augments two existing longitudinal cohort studies of preclinical and clinical AD, the Wisconsin Registry for Alzheimer’s Prevention and the Wisconsin Alz- heimer’s Disease Research Center, with rich phenotypic data from blood, cerebrospinal fluid (CSF), imaging, lifestyle questionnaires, and neuropsychological testing. The overall objective is to measure plasma and CSF metabolomics in additional longitudinal samples and use sophisticated data analysis approaches to establish the timing, trajectory, and etiology of metabolite changes in the disease process. The central hypothesis is that changes in metabolites are influenced by genetics and lifestyle and occur at distinct stages of AD pathology. The rationale for the proposed research is that a better understanding of the timing, trajectory, and etiology of AD- related metabolomic changes is critical to prevent (e.g., lifestyle interventions), diagnose (metabolomic bi- omarkers), and treat (new therapeutic targets) the disease. The central hypothesis will be tested by executing the following specific aims: 1) determine the timing and trajectory of plasma and CSF metabolites throughout the AD process using sophisticated longitudinal modeling approaches, 2) integrate genomics and metabolomics to determine which AD-associated metabolites are in the causal pathway to AD using Mendelian randomization analyses, and 3) determine which AD-associated plasma and CSF metabolites mediate the relationships be- tween AD-associated lifestyle factors and AD-related outcomes. At the conclusion of this project, expected out- comes include: 1) identification of metabolites/pathways that are precursors to AD pathologic changes and may be therapeutic targets versus those that change in the early stages and can be used as early biomarkers versus diagnostic/prognostic metabolites that are markers of more advanced disease, 2) identification of metabolites that are in the causal pathway and may inform therapeutic targets, 3) a better understanding of mechanisms linking metabolic and vascular disease processes with AD, and 3) identification of metabolites linking lifestyle factors to AD risk that can inform future intervention trials and clinical practice by identifying more specific be- havior changes and providing biomarkers of biologic change to monitor the effectiveness of interventions with a shorter period of follow up.

NIH Research Projects · FY 2025 · 2016-08

PROJECT SUMMARY The Wisconsin Head and Neck SPORE is designed to promote translational laboratory and clinical research to improve overall outcome for patients with Head and Neck Cancer (HNC). This highly collaborative research links basic scientists with HNC clinicians to advance novel treatment strategies for this complex cancer population. These patients bear a disproportionate burden from their cancers based on the critical anatomic location of the disease for which treatment can compromise speech, swallow, and breathing function, in addition to creating significant alterations in physical appearance and capacity for social interaction. Efforts to improve cure rates must be carefully balanced with efforts to reduce treatment toxicity to enable enhanced overall quality of life for patients. The broad objectives of this SPORE are to: 1) Promote multidisciplinary translational research in HNC, 2) Improve overall survival and quality of life for patients with HNC, 3) Incorporate new predictive models to test novel HNC treatment strategies, 4) Improve understanding of how immune modulation can augment conventional and experimental treatment responses in HNC, 5) Translate promising new molecules developed at the University of Wisconsin and from Industry through preclinical testing and into HNC clinical trials. The Wisconsin HN SPORE has designed three primary research projects. Project 1 will combine targeted radionuclide therapies (TRT) with immune checkpoint inhibition (ICI) to stimulate enhanced HNC response profiles culminating in a Phase I clinical trial. Project 2 builds a powerful patient-specific bioengineered HNC model system from patient cells that incorporates components of the tumor microenvironment to more accurately predict HNC patient treatment response. The feasibility of using treatment response data from the model to inform postoperative radiation therapy will be tested in a clinical pilot study. Project 3 examines dual targeting of critical receptor tyrosine kinases Axl and MerTK to mediate changes in the immune microenvironment and thereby augment tumor response in HNC patients. The Wisconsin SPORE will support this research with three Cores (Administrative, Pathology and Biostatistics), a Career Enhancement Program and a Developmental Research Program.

NIH Research Projects · FY 2026 · 2016-08

1 PROJECT SUMMARY 2 The overarching goals of the African Americans Fighting Alzheimer’s in Midlife (AA-FAIM) renewal are to 3 promote timely and valid detection of Alzheimer’s disease (AD) and related dementias (ADRD) in Black 4 populations and to increase inclusion of Black adults in ADRD research. Being racialized as Black in the US is 5 associated with twice the risk of cognitive decline and ADRD relative to non-Hispanic whites. Yet, because Black 6 Americans are substantially under-recruited and under-represented in ADRD research, gaps remain in our 7 understanding of the generalizability of prevailing theories of preclinical AD pathophysiology (e.g. the amyloid 8 hypothesis) to Black patients. To address these gaps, we must obtain essential biomarker data from Black 9 participants and improve inclusion of Black participants, examining recruitment/retention with scientific rigor. In 10 Aim 1, we will explore the relevance of preclinical amyloidosis in predicting cognitive decline in a Black cohort, 11 comparing models considering roles of (a) preclinical AD pathology, (b) vascular risk factors, (c) psychosocial 12 factors, and (d) cognitive markers in predicting cognitive decline in a cohort of 400 Black adults. Aim 2 will 13 examine the association between plasma Aß42/40 and Amyloid PET positivity (PET A+) and test the 14 chronicity/EAOA (estimated age of amyloid onset) model of preclinical AD in Black participants. We will 15 investigate associations between plasma Aß42/40 and concurrent PET A+, and assess whether longer amyloid 16 chronicity and/or earlier EAOA are associated with accelerated cognitive decline. Aim 3 examines factors 17 associated with successful enrollment and retention. We will assess patterns of perceptions and beliefs relative 18 to an emerging model of participant engagement, and explore model constructs’ efficacy in predicting 19 prospective participation decisions. With renewed investment in the AA-FAIM cohort, we will continue 20 contributing to an emerging understanding of preclinical ADRD in a minoritized population. Moreover, we can 21 further leverage our participants’ contributions by partnering with teams seeking access to these unique data.

NIH Research Projects · FY 2025 · 2016-07

ABSTRACT – MIDUS OVERALL PLAN The Midlife in the U.S. (MIDUS) national longitudinal study has been ongoing since 1995. MIDUS is the only national study focused on midlife with a wide age expanse at baseline. MIDUS also has unusual depth in its psychosocial, biomarker, genomic, and neuroscience assessments, thus permitting a focus on neurobiological mechanisms and pathways through which sociodemographic and psychosocial factors influence morbidity and mortality. This application aims to conduct a 2nd wave of data collection on the MIDUS Refresher sample (MR2) as well as a 4th wave of data collection on the Core sample (M4), including all projects in both. Four projects (Survey, Daily Diary, Biomarkers, Genomics) are part of this U19 competing continuation application, which is linked with an Ancillary U01 application on Alzheimer’s Disease (AD) and Related Dementias (ADRD) that will examine midlife precursurs to cognitive decline, emotion regulation, brain aging, and their interplay. Much of that prior work is also longitudinal, although it now includes new ADRD neuroscience and biomarkers. Overall, the proposed activities involve over 5,200 U.S. adults that will be supported by an Administrative Core responsible for orchestrating cross-project data collection and delivering high-quality, well-documented data; a BioCore that ensures quality control in biomarker data collection and offers guidance on use of biomarkers; and a Statistics Core that provides workshops on multiple topics (modeling longitudinal change, using genomic data, linking MIDUS to other datasets). Recurring scientific themes in the proposed science are health inequalities and racial disparities examined with the rich biopsychosocial data available in MIDUS, including wide-ranging assessments of stress exposures across multiple waves, thereby providing indicators of cumulative adversity. MIDUS is also known for its comprehensive assessments of psychosocial and behavioral protective factors, thereby advancing research on resilience in the face of challenge. Aging on a changing historical stage is another key theme in MIDUS exemplified by a past focus on hardships of the Great Recession, and going forward, a new parallel focus on hardships of the COVID-19 pandemic. In terms of scientific engagement, MIDUS is the most frequently downloaded study at the National Archive of Computerized Data on Aging (NACDA). Widespread usage from the scientific community (26,000+ public users) has culminated in 1,617 publications covering 38 substantive domains. Underscoring the momentum behind MIDUS, more than half of these products are journal articles published during the current funding cycle (2016-present).