University Of Wisconsin-Madison

NIH Research Projects · FY 2024 · 2012-08

Abstract Spatial RNA-sequencing has emerged as a revolutionary tool that allows us to address scientific questions that were elusive just a few years ago. Specifically, the spatial RNA-sequencing technology has the potential to revolutionize studies of tissue structure and function in health and disease. However, much of the potential has yet to be realized as statistical methods to analyze spatial RNA-seq data are lacking. For many types of analyses, the methods currently in use obscure and, in some cases, distort biological signals. A number of statistical and computational challenges must be addressed to prevent inaccurate conclusions, and to optimize novel discovery. This proposal addresses those challenges. In particular, while the technology is powerful, it is not without error; and considerable contamination exists in spatial RNA-seq data. We propose methods to remove this contamination and thereby ensure robust and accurate downstream inference. We also propose statistical methods to adjust for technical variability induced by differences in sequencing depth. By reducing technical variability, these methods will improve the power with which signals of interest can be studied. Finally, we propose methods for characterizing changes in the dependence structure of sets of genes. These types of methods are required to improve our understanding of how coordinated changes in genes affect tissue structure and function in health and disease. Taken together, successful completion of this project will help to ensure that maximal information is obtained from powerful spatial RNA-seq experiments.

NIH Research Projects · FY 2025 · 2012-08

Dysregulation of lipid metabolism is strongly associated with age-dependent retinal diseases including age-related macular degeneration (AMD) based on genetic and epidemiological studies. However, the roles of dysregulated lipid metabolism in the development and progression of age-dependent retinal diseases remain largely unknown. Mouse models showing impairment of lipid metabolism and cellular homeostasis in the retina provide excellent tools to study how dysregulated lipid metabolism impacts retinal health and lead to age- dependent retinal diseases. We identified one such mouse strain harboring a mutation in transmembrane protein 135 (Tmem135) that displays signs of accelerated aging in the retina as well as pathologies observed in AMD including retinal pigment epithelium (RPE) cell abnormalities, inflammation and photoreceptor cell degeneration. We found that mitochondrial dynamics are dysregulated in Tmem135 mutant mice. In the previous funding period, we further identified the role of TMEM135 in lipid metabolism in the retina. Our lipidomics data showed reduced DHA levels in Tmem135 mice and indicated that TMEM135 has a role in DHA export from peroxisomes. We also found that TMEM135 is critical for the regulation of peroxisomal number and lipid metabolism in the retina. The number of peroxisomes is correlated with mitochondrial morphology and function in Tmem135 mutant mice and mice overexpressing Tmem135 (Tmgm135 TG), suggesting close interaction between these organelles. Moreover, the Tmem135 mutation changes expression of genes associated with lipid metabolism in mouse eyecups, which are similar to genes differentially expressed in RPE/choroid of AMD patients. Based on these findings, we hypothesize that “Regulation of peroxisomal functions through TMEM135 is essential to maintaining the normal function and integrity of RPE and photoreceptor cells, dysregulation of which leads to age-dependent retinal disease phenotypes.” In this renewal proposal, we will investigate how TMEM135 regulates lipid metabolism, mitochondrial function and retinal function through its role in peroxisomes and DHA synthesis. Specifically, we will 1) test the hypothesis that decreased DHA levels in Tmem135 mutant mice are responsible for the retinal abnormalities, 2) determine the contributions of fatty acid synthesis pathways on Tmem135 mutant retinal pathologies, and 3) test the role of peroxisomes in mitochondrial homeostasis in the RPE. Successful completion of this project will reveal the previously unknown role of Tmem135 in age-dependent changes of retinal cells, and identify factors involved in those processes, which may lead to novel supplementation or treatment options for aging and age-related diseases in the retina.

NIH Research Projects · FY 2026 · 2011-09

PROJECT SUMMARY/ABSTRACT Chronic myelomonocytic leukemia (CMML) is a devastating cancer with an urgent and unmet need for effective therapies (median survival: ~28 months). Approximately 30% of CMML cases evolve to acute myeloid leukemia (AML) soon after their initial diagnosis, contributing to the poor prognosis of CMML patients. During the prior funding period, we collaborated with the International MDS/MPN Consortium and demonstrated that concurrent NRAS and ASXL1 mutations define a population of CMML patients with shorter leukemia-free survival than those with ASXL1 mutations only. Based on our human data, we characterized NrasG12D/+; Asxl1-/- (NA) mice that model CMML patients with concurrent NRAS and ASXL1 mutations. NA mice developed CMML with accelerated progression and in ~50% of these mice CMML transformed to AML (secondary AML, sAML). NA leukemia cells exhibited hyperactivation of MEK/ERK signaling and increased global level of H3K27Ac, a histone mark bound by bromodomain and extra-terminal domain (BET) proteins for gene transcriptional activation. Upregulation of AP-1 transcription factors (TFs) mediated the overexpression of PD-L1 and CD86, two inhibitory immune checkpoint ligands, and helped establish a suppressive immune microenvironment in NA-sAML recipients. Combined inhibition of MEK and pan-BET proteins led to downregulation of AP-1 TF expression, partial mitigation of the suppressive immune microenvironment, enhancement of CD8 T cell cytotoxicity, and prolonged survival in NA-sAML mice. Based on our preliminary results, we hypothesize that Asxl1-/- and oncogenic Nras cooperate to accelerate CMML and promote its transformation to AML via reprogramming the immune microenvironment, which includes but may not limit to T cells. Moreover, immunomodulatory agents may further improve the therapeutic benefits of combine MEK and BET inhibition through establishing durable anti-leukemia activities in immune cells. In this renewal application, we propose the following two aims to test our hypothesis: 1) To identify the molecular and cellular mechanisms underlying the dysregulation of immune microenvironment in in NA mice; and 2) determine whether immunomodulatory agents further improve the therapeutic effects of combined MEK and BET inhibition in NA mice. Together, our proposed studies will not only provide fundamental perspectives on disease mechanisms but also test hypotheses that could potentially lead directly to potent and efficacious novel therapies for treating CMML and transformed AML.

NIH Research Projects · FY 2026 · 2011-08

Abstract (Summary) The primary objective of the Molecular and Environmental Toxicology Summer Research Opportunities Program (MET-SROP) is to accelerate the educational pipeline of underrepresented students and train the next generation of researchers in the environmental health sciences. The overarching idea of the MET-SROP is that by providing an opportunity to experience a significant environmental toxicology laboratory research project with strong mentorship and employ modern technologies to understand cellular and molecular mechanisms of toxicity, we can bring down the barriers that prevent many young scientists from choosing such a career path and simultaneously enhance their likelihood of success. Building on over ten years of success, the program aims to support seven undergraduate trainees from underrepresented backgrounds each summer and provide research experiences that reflect modern approaches to understand cellular and molecular mechanisms of toxicity. Unique program elements include: i) rigorous hands-on training in basic laboratory research coupled with ii) introduction to a didactic foundation of environmental health sciences, and iii) diverse exposures to future career opportunities and tools for successful advancement beyond undergraduate training. Housed at the University of Wisconsin-Madison Campus, trainers are experts in foundations of toxicology, molecular approaches to chemical toxicity, genetics, data sciences, and epidemiology. Trainees receive extensive laboratory experience working alongside graduate students and fellows. This laboratory training is combined with a ten-week program including didactic training, career development and mentoring. The parallel “Didactic” track program will begin with an interactive overview of toxicology and its relevance to other environmental health sciences including environmental regulation, risk assessment, epidemiology, community health and health equity. This foundation is essential for supporting student success in lab work and the ability to see the impact of research beyond the laboratory. The didactic session also includes a three-week minicourse in data science including training in R-programming for analyzing toxicology and human health data. The third SROP component, referred to as “Science Life,” includes several minisymposia throughout the ten weeks that include laboratory tours, and talks by leading experts from UW Madison, and former trainees known in the environmental health sciences workforce, from both government and industry. Finally, SROP trainees participate in a variety of campus events to interact with and build connections with other summer program trainees from diverse backgrounds. This networking is also complemented by numerous opportunities to engage with other graduate students and alumni through the didactic and science life tracks. Since its inception, the MET-SROP aims to welcome and support students from communities traditionally underrepresented in STEM and provides them with a rewarding summer experience from which to advance their environmental health sciences career.

NIH Research Projects · FY 2025 · 2011-07

Project Summary/Abstract The discovery of new potential drugs and biological tools is necessarily limited to the conveniently-available chemical space. Historically, this space has been dominated by biaryls because their synthesis is reliable and amenable to the needs of medicinal chemistry, where a core structure is diversified and built outwards by iterative rounds of synthesis. Recently, cross-electrophile approaches show promise for providing convenient access to more C(sp3)-rich molecules, a feature predicted to improve the odds of success in drug development. Despite recent advances, cross-electrophile coupling of aryl electrophiles with alkyl electrophiles has yet to realize its considerable potential. This is because many of the most abundant pools of substrates have very different reactivity. We propose to develop new strategies in cross-electrophile coupling that address these challenges and are adapted to modern medicinal chemistry approaches. This program's long-term goals are the development of methods for the selective cross-coupling of every major class of electrophile and the discovery of the fundamental properties that control selectivity and reactivity. In the proposed grant, a team of graduate students and a postdoc will build upon the advances of the previous grant period to develop protocols to cross- couple starting materials sourced from the largest substrate pools (organic chlorides, alcohols, amines, and carboxylic acids), access more hindered C(sp2)–C(sp3) bonds, and shed light on how the nature of the coupling partners and ancillary ligands govern success. Our guiding hypothesis is that these challenges can be addressed by a combination of mechanistic studies, mechanism-guided tuning of catalysts and activating agents, and an optimization approach that focuses on a collection of substrates rather than a single substrate pair. The specific aims of this proposal are to: (1) develop protocols for C(sp2)–C(sp3) cross-electrophile coupling between the largest pools of aryl, vinyl, and alkyl substrate pools; (2) address the challenge of forming C(sp2)–C(sp3) bonds by developing new catalysts based upon nickel and cobalt; (3) study how ancillary ligands influence the stability and reactivity of arylnickel(II) complexes to both improve catalytic reactions and to enable new types of stoichiometric reactions for medicinal chemistry applications, such as DNA-encoded libraries. The approach is innovative because cross-electrophile coupling is less studied than other cross-coupling methods and the proposed mechanistic studies will shed light on these little-understood processes. The proposed research is significant because the chemistry is increasingly important to industrial and academic chemical synthesis and the development of nickel chemistry has outpaced our understanding.

NIH Research Projects · FY 2025 · 2011-06

Project Summary The purpose of this program is to provide a 12-week mentored research experience for veterinary students at the University of Wisconsin-Madison. The traditional veterinary curriculum lacks formal training in performance of research: therefore, this summer research exposure experience is an essential way to expose DVM students to the research process within a tier-one research institution and encourage them to consider research-oriented careers. This program has been integral to our ability to recruit prospective veterinary medical students with research interests to our DVM professional program at the University of Wisconsin- Madison. It has also been fundamental to our success in increasing the number of minority veterinary students pursuing research training, informing veterinary medical students of careers in research, and exposing veterinary students to important discussions related to research ethics, responsible conduct of research, and the skills and tools required for successful careers in research. The over-arching objective of this program is to introduce veterinary medical students to research. They gain a working knowledge of how to identify a problem of significance to the health and welfare of animals and/or humans, learn how to develop a hypothesis to test focused questions, build a plan to identify and validate appropriate techniques to pursue, analyze and report data. The students undergo workshops to promote excellent science communication in both oral and written formats. Trainees participate in regularly scheduled laboratory meetings and weekly seminars designed specifically for these goals within the 12-week summer timeline. Trainees are expected to present the results of their research at the summer veterinary research scholars symposium (typically the NIH-Boehringer Ingelheim Veterinary Scholars Program Symposium held annually in August) or at seminars or symposiums held at the University of Wisconsin School of Veterinary Medicine. The University of Wisconsin in general, and the School of Veterinary Medicine in particular, provides an ideal environment to support short-term research training for veterinary students. The proposed program is designed to integrate with additional graduate research training programs within the School of Veterinary Medicine to provide a continuum of support for veterinary students that are inspired to pursue advanced research training.

NIH Research Projects · FY 2025 · 2010-06

HMPV is a major cause of lower respiratory infection (LRI) in children worldwide, second only to respiratory syncytial virus (RSV). Although nearly all people are infected with HMPV during childhood, immunity to HMPV is incomplete and re-infections occur throughout life. Hospitalization with HMPV is more likely in persons with underlying conditions such as asthma, chronic obstructive pulmonary disease, HIV, or prematurity. There are no approved antiviral therapies or vaccines. Host or viral determinants of virulence are not known for HMPV. One limitation of many prior studies is that most HMPV research uses one of a few lab-adapted strains that cause minimal disease in mice. We have identified clinical isolates of HMPV that cause severe and fatal disease in mice, providing tools to elucidate mechanisms of pathogenesis. Our preliminary data show that virulent HMPV potently induces types I and III interferon (IFN), and that these cytokines exhibit discordant roles. IFN is critical to initiate and shape adaptive immune responses but can contribute to disease; we aim to define the contribution of types I and III IFN in HMPV. HMPV inhibits type I IFN responses including STAT1 and STAT2 phosphorylation by an unknown mechanism, and HMPV lacks the paramyxovirus V protein or NS1/NS2 of RSV. Several HMPV proteins have been implicated, including G, M2-1, P, and SH. Published data from our group and others suggest SH mediates immune inhibition but a mechanism or specific SH-host protein interaction is unknown. The contribution of other HMPV proteins to virulence has not been defined. We propose to use in vitro and in vivo approaches to address these knowledge gaps. In Aim 1, we will define the contributions of type III IFN (IFN-λ) to HMPV immunity and pathogenesis using global and conditional knockout mice. Aim 2 proposes to identify the cellular target(s) of SH, define interacting domains, and discover the molecular mechanisms of innate immune inhibition by HMPV. We will use transient transfection of tagged mutant SH proteins and viruses with SH mutations in cell and mouse experiments. In Aim 3, will use reverse genetics developed in the lab to generate chimeric viruses and identify viral protein determinants of virulence using established mouse models. Our preliminary data suggest an important role for IFN-λ in HMPV immunity, confirm innate immune inhibition by HMPV SH, and demonstrate the strength of the avirulent and virulent strains to identify viral determinants of virulence. The results of the proposed research will clarify mechanisms of HMPV pathogenesis and provide a blueprint for potential therapeutic avenues and strategies for viral attenuation for vaccines. The findings are likely to be relevant to other respiratory viruses.

NIH Research Projects · FY 2025 · 2009-07

PROJECT SUMMARY/ABSTRACT Since 2009, the long-term goal of the UW Voice Research Training Program has been to foster the development of translational research skills in future leaders in the field of voice science. We provide promising predoctoral and postdoctoral fellows with highly interactive and multi-disciplinary training experiences that are unfettered by conventional departmental barriers, and actively facilitate their development as independent scientists. Fellows partake in comprehensive laboratory, translational, and clinical research experiences, as well as exposure to curricula in clinical trials, hypothesis-based research, management, ethics, and data analysis. Further, the training program offers medical students a short-term summer research opportunity to encourage future clinicians to become physician-scientists. The program capitalizes on the abundant university resources, including the UW CTSA Institute for Clinical and Translational Research, thereby creating an outstanding voice training program that is not matched elsewhere. Our pool of experienced, extramurally funded trainers, from a variety of disciplines including otolaryngology-head and neck surgery, medicine, human oncology, biomedical and chemical engineering, bacteriology, and communication sciences and disorders incorporates effective assessment processes, a plan to promote diversity by recruiting and retaining both women and minorities, and a solid plan for training in the responsible and ethical conduct of research. This renewal application, based on our immense success over the last 14 years, requests an additional five years of funding for 5 predoctoral, 3 postdoctoral, and 2 short-term fellows each year. An important metric of success of our past performance to date is that 31 predoctoral and postdoctoral fellows have completed the program (6 fellows are currently appointed) and have progressed onto competitive postdoctoral positions and academic, industry, or government research careers. We have had 24 short-term medical students with 2 enrolled for summer 2023. Other indicators of success of the program include 17 NIH F31 and F32 fellowship awards to our trainees, other NIH funding (Loan Repayment, K23, K99, R21 and R01s) received to past trainees, in addition to countless other awards, grant funding, and scholarships. For each fellow, the UW Voice Research Training Program has a proven track record of creating a dynamic research experience with an outstanding collaborative research team. Renewal of this training program will ensure the continued excellence and engagement of the next generation of voice scientists.

NIH Research Projects · FY 2025 · 2008-09

Structural mechanism of diabetic amyloid formation Abstract 11.6% of the U.S. population is afflicted by type 2 diabetes. It starts as insulin resistance, but ultimately the pancreatic -cells that make insulin fail, resulting in overt diabetes. -cell failure correlates with aggregation of a hormone called the human islet amyloid polypeptide (hIAPP or amylin) into amyloid plaques in the islets of the -cells. Surprisingly, the amyloid fibers themselves are not cytotoxic. Many researchers believe that the toxic species are oligomers of hIAPP that interfere with receptor mediated processes, cause inflammation, or permeabilize the membrane. The oligomer hypothesis is strengthened by recently reported antibodies that bind specifically to oligomers and stall type 2 diabetes in animal models. As a result, there is much interest in elucidating the structures of oligomers and understanding the mechanism by which they form. However, characterizing kinetically evolving proteins is challenging and so little structural information exists about these transient oligomers. We invented a technology, rapid-scan 2D IR spectroscopy, that enables us to monitor the structure of hIAPP as it aggregates. We discovered an oligomeric intermediate with a parallel -sheet in the FGAIL region and an -helix at the N-terminus. We observed this “FGAIL oligomer” in 4 different mammalian species known to contract type 2 diabetes, strengthening our hypothesis that this intermediate is a key player in the disease. Most importantly, we realized that we could trap the oligomer with a few benignly placed mutations. Our trapped oligomers are nearly as toxic as wild-type hIAPP, but persist in vitro for days rather than hours. Because they are stable for so long, we could perform 2D/3D NMR and generate the first structural model of an hIAPP oligomer. Simultaneously, we created a transgenic mouse model that expresses our oligomer, thereby linking our in vitro oligomer to pathophysiology. In this proposal, we capitalize on our new strategy for studying the structures and mechanisms of hIAPP oligomers. Aim 1 refines our structural model, Aim 2 extends our approach to a hereditary missense mutation that causes early onset-diabetes, and Aim 3 tests our in vitro hypotheses with matching transgenic mouse models. We seek to understand hIAPP aggregation from a fundamental perspective and link our in vitro structural mechanisms to in vivo physiology. We believe that our approach is unique to the structural biology community, providing much desired structural and mechanistic information about hIAPP aggregation and its relationship to type 2 diabetes.

NIH Research Projects · FY 2025 · 2008-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The goal of the Childhood Diabetes and Adiposity Research Training program (C-DART) is to prepare post-doctoral physician and PhD scientists for productive careers in clinical, translational, or basic research in prevention or treatment of type 1 and 2 diabetes and other metabolic consequences of obesity. The training program has evolved during the current funding period by augmenting research training of MD physician scientists with successful recruitment of an outstanding PhD trainee. C-DART has also applied Team Science Learning Points to formally, critically evaluate and modify training program components. Insulin resistance, strongly associated with ectopic fat deposition, is a major health problem in children, and rates of both type 1 and type 2 diabetes continue to rise in the pediatric population. Accordingly, C-DART trainees begin a research career ranging from molecular/cellular study of pancreatic islet biology and insulin resistance to whole body pathophysiology of childhood adiposity to prevention/treatment of childhood obesity in ethnically diverse youth to, finally, optimizing use of rapidly advancing technological and pharmacological therapeutics for childhood obesity, Type 1, and Type 2 diabetes. Training objectives include 1) establishing quality and productive research projects, 2) developing a record of peer-reviewed publications, and 3) transitioning to successful research- focused academic careers for 80% of graduates, with most of those submitting proposals for extramural funding during T32 tenure such as NIH K series (or similar) to continue their research careers. To achieve these objectives, a research Capstone Certificate curriculum developed in collaboration with the NIH-funded UW Institute for Clinical and Translational Research (UW-ICTR) provides training in research techniques, statistics and study design, responsible conduct of research, and scientific writing and presentation skills. The rising burden of diabetes and other obesity-related metabolic disorders, the strong 15-year record of accomplishment of our T32-supported trainees, and a scarcity of young basic and clinical science researchers with childhood obesity and diabetes training prompt us to seek continued funding for the C-DART program. The importance of sustaining successful pediatric endocrinology research training programs like C-DART is magnified by a serious shortage of young physician scientists committing their careers to care of children with obesity and diabetes. Since the inception of thisT32, 12 of 14 fellows obtained, upon graduation or soon after, faculty appointment opportunities that include research activities and innovative program development in pediatric diabetes, fitness, and obesity at academic medical schools. During the proposed renewal funding period, C-DART will continue utilizing existing resources and new strategies to recruit the most qualified trainee candidates. The program will continue to build collaborations with new potential mentors engaged in innovative and cutting-edge science to provide state-of-the-art mentored training for productive research careers in childhood diabetes, insulin resistance and obesity.

NIH Research Projects · FY 2025 · 2008-06

Project Summary Candida commonly adhere to medical devices, flourishing as a biofilm. A clinical hallmark of biofilm infection is profound antifungal resistance. As effective therapies are not available for these life-threatening infections, understanding how the cells survive existing drug therapies is desperately needed. The extracellular matrix, a distinguishing feature of biofilms, has been linked to drug resistance. This proposed investigation builds logically upon progress in the last funding period, including; the discoveries that 1] key matrix components are delivered via biofilm distinct extracellular vesicle (EVs) as cargo components, 2] exogenous EV add-back assays allow delineation of vesicle cargo constituent roles, 3] ESCRT (endosomal sorting complexes required for transport) pathway components partially impact biofilm EV production and cargo, and 4] a novel antifungal, turbinmicin, inhibits biofilm EV production and matrix production, rendering the cellular community susceptible to antifungals. 5] We further identified a few genetic components necessary for production of these EV and biofilm roles for select vesicle cargo components. However, the functions of the majority of vesicle cargo constituents and the regulatory components of these biofilm pathways remain undefined. Our recent progress helps to solve this mystery. We recently discovered TF mutants with three distinct EV and biofilm resistance and matrix phenotypes. We hypothesize that investigation of these loss of function TFs mutants and their effectors will 1] identify the regulatory network for biofilm EV production and cargo packaging of EVs and 2] uncover roles for EV cargo in biofilm biology, including drug-resistance. Our hypothesis is based upon five main findings. First, we identified a group of TF mutants exhibiting both antifungal susceptibility and extracellular matrix defects, consistent with a matrix protection resistance mechanism. Second, we discovered a subset of drug-susceptible mutants displaying a profound reduction in biofilm EV production. Importantly, the matrix resistance phenotype of these TF mutants is restored via administration of WT EVs. Third, another group of drug-susceptible TF mutants exhibit changes in vesicle cargo but preserved vesicle production. The drug-susceptible phenotype is also reversed for these mutants by addition of WT EVs. Fourth, we find a group of TF mutants that produced elevated quantities of biofilm extracellular vesicles. And fifth¸ exposure of biofilms to turbinmicin reduces not only EV quantity but EV cargo constituents. Our immediate objectives are 1] to define the Candida regulatory pathways that govern vesicle delivery and maturation of the matrix-resistance mechanism and 2] to discern the genetic effectors responsible for production and delivery of these virulence constituents. We seek to test our EV biofilm hypotheses utilizing these three distinct functional TF mutant groups, turbinmicin as a novel pharmacologic tool, and our EV and biofilm assays.

NIH Research Projects · FY 2025 · 2008-06

Project Summary Children with cerebral palsy (CP) have a range of communication ability profiles. The majority have the speech motor disorder, dysarthria, which almost always results in reduced speech intelligibility. Heterogeneity among children with CP poses challenges for the study of speech and language development, necessitating the use of prospective longitudinal methods wherein each child can serve as their own control. We began such work 15 years ago, following 90 children with CP, with the long-term goal of generating theoretically driven, empirically validated, longitudinal models of speech and language development that can be used to predict outcomes, guide treatment decisions, and test interventions for children with CP. To date, we have developed and validated prospective, data-driven models of speech and language development, and created a clinical speech-language profile group paradigm for children with CP. Our research has resulted in the first published growth curves for speech intelligibility, and language comprehension, leading to the ability to predict later speech and language outcomes with a high level of accuracy based on performance at only 3 years of age. Our data have revealed that children with CP have a protracted developmental timeframe for the acquisition of speech, with many still making intelligibility improvements through the age of 15 years. Despite major gains in our understanding of speech and language development and in the application of this knowledge to clinical decision making, progress is limited by two key barriers, addressed in this application: a.) The upper limits of speech and language development after 15 years of age in CP have not been studied; therefore, we are unable to generate comprehensive growth models that encompass all of development and include young adult outcomes. b.) Treatment options for improving speech intelligibility in children with CP are limited and have primarily focused on remediating speech subsystem deficits. Interventions focused on using augmentative and alternative communication (AAC) strategies to supplement speech have not been examined in children, but findings from adult CP studies are promising. In this renewal, we will collect new longitudinal data on our existing cohort of youth with CP through the age of 20 years. We will also study 100 new children with CP in a Phase I speech supplementation intervention study. Aims are: 1.)To quantify longitudinal change in speech and language development between the ages of 2 and 20 years in children with CP. 2.)To develop and test basic elements of a speech supplementation intervention to improve intelligibility in children with CP (Phase I behavioral clinical trial). This work will complete the development and validation of longitudinal models of speech and language growth in children with CP to 20 years of age. Results will enable the prediction of outcomes for children with CP between 2 and 20 years, which will have direct implications for intervention decision-making. Our Phase I intervention trial using speech supplementation strategies to improve intelligibility in children with CP will lay the foundation for future Phase II and III clinical trials.

NIH Research Projects · FY 2025 · 2007-07

Summary: Veterinarians are at the forefront of the ‘ONE HEALTH’ strategic framework, which facilitates collaborations among human medicine, veterinary medicine and ecosystem health that are of worldwide benefit. The vital importance of veterinarians at the human-animal interface cannot be overstated, as graphically illustrated by the tragic repercussions of the ongoing COVID-19 pandemic, and that 6 of 10 human infectious diseases are zoonotic in nature. Reports from the National Academy of Sciences and the strategic plan of the NIH Office of Infrastructure Programs (ORIP) emphasized the national need for veterinarians trained in biomedical research to effectively deal with effects of climate change on animals and public health, thwart the emergence of zoonotic diseases, and promote human and animal health. We will address the national need for veterinary biomedical scientists through these specific aims: Aim 1: Develop the next generation of independent veterinary medical researchers equipped with skill sets in innovative and critical thinking, problem solving, robust experimental methodologies and rigorous data interpretation. Aim 2: Continually innovate professional development opportunities for veterinary medical scientists in grantsmanship, science communication, team science, evidence-based mentor and mentee practices, and forward-thinking leadership. The School of Veterinary Medicine (SVM) and the University of Wisconsin-Madison offer an exceptional research training environment and are ideally positioned to provide outstanding training to graduate veterinarians in cutting-edge biomedical research. The excellent research training offered by this program is highlighted by the (1) broad multidisciplinary portfolio of an outstanding group of trainers with a history of sustained funding, productivity and commitment to graduate training; (2) nationally ranked multidisciplinary research training program (Comparative Biomedical Sciences PhD program) with major strengths in infectious disease and immunology, physiologic basis of health and disease, and developmental biology/regenerative medicine; (3) proven ability to recruit and train a diverse group of outstanding veterinarians in biomedical research; (4) strong research ethics and professional skills course and access to an array of career/professional development programs; (5) commitment to minority student recruitment; and (6) strong institutional and administrative support from the SVM and campus. We request funds to support four postdoctoral DVM trainees per year to pursue PhD training in a 5-year period. The overall goal is to train veterinary biomedical scientists in hypothesis-driven science who can assume leadership roles in biomedical research, academic instruction, industry innovations, and government service in the 21st century.

NIH Research Projects · FY 2026 · 2007-05

Abstract PROJECT SUMMARY/ABSTRACT: Population health challenges in the United States are large and historically rooted, begin early in life, and often persist throughout the life course. The Health and Development Research Scholars Program (HDRS) is an interdisciplinary postdoctoral training program located in the Department of Population Health Sciences at the University of Wisconsin School of Medicine and Public Health. It is designed to support the career development of population health and development researchers and has two primary goals: 1) to develop a cadre of researchers who will work to advance knowledge of the causes, consequences, and effective mitigation of health challenges experienced by many populations early in and throughout the life course; and 2) to attract, retain, and support the development of a highly qualified set of population health investigators as they embark on research careers dedicated to improving health. To achieve these goals, we will actively recruit a cadre of scholars who are committed to careers in health research and provide them with two or three years of training and support. We will recruit from doctorate programs scholars who are ready to build on the methods of their graduate disciplines, cross boundaries, and draw from content, theory, and methods of other fields. We have assembled an exceptional group of core faculty with expertise across medical, population health, social/behavioral, and basic sciences. There are five training components to the training program: i. a multidisciplinary mentor team; ii. a weekly HDRS seminar and the opportunity to attend other seminars on campus; iii. coursework, workshops, and optional degree programs; iv. mentored research; and v. professional development activities. Through this interdisciplinary training program, we will build key competencies and provide professional mentoring to facilitate the success of our scholars as independent population health researchers. Scholar activities will be tracked through quarterly progress reports, biannual mentor team meetings, and an annual formal evaluation of each scholar. These assessments will enable the Program Director and mentor team to provide timely guidance and identify strategies to ensure productivity and achievement of career goals.

NIH Research Projects · FY 2025 · 2006-09

Project Summary The decline in synaptic plasticity with age is thought to impose severe constraints on the recovery from amblyopia in adults. Although this developmental loss was previously thought to be irreversible, our previous work established that robust plasticity can be reactivated in the adult visual cortex by dark exposure (DE) followed by light reintroduction (LRx). We outline a series of experiments to delineate the cellular and molecular mechanisms that couple DE/LRx to the rejuvenation of synaptic plasticity in the adult visual system. We propose that DE induces homeostatic changes in the composition of synaptic NMDARs in the primary visual cortex, which lowers the threshold for Hebbian plasticity. However, the response to DE over- compensates for the loss of visual input and results in neuronal hyper-excitability. The threshold for activation of perisynaptic proteolysis by LRx at thalamocortical synapses is also lowered by DE. The response to LRx reduces feed-forward excitation to the cortex and is mediated by rapid plasticity of presynaptic function.

NIH Research Projects · FY 2026 · 2006-05

PROJECT SUMMARY/ABSTRACT Feeding behavior is critical for animal survival, and is also a fundamental aspect of energy homeostasis. This process is regulated by highly complex neurochemical pathways involving a multitude of neuropeptides and biogenic amines. Despite decades of work on individual neurochemical systems, the general organizational principles underlying neuromodulation are still poorly understood. This is mainly due to the fact that modulation of neural circuit has so far been studied primarily one modulator at a time without the knowledge about co-modulation of networks. The latter information would require the development of sensitive and selective analytical tools to precisely identify these low abundance endogenous signaling molecules and accurately measure their behaviorally-relevant concentrations in a complex microenvironment. Our proposed research aims to address this critical knowledge and technological gap by developing new bioanalytical methods to elucidate the complex identities and functional roles of neuropeptides in food intake via combined mass spectrometric and physiological approaches. We employ the crustacean stomatogastric nervous system, cardiovascular system, and its associated neuroendocrine organs as a test-bed for technology development and validation due to the unique advantages and biological significance of this model system. In parallel, we aim to translate our technology development for neuropeptide discovery and analysis to the mammalian central nervous system. To this end, we propose to focus on key brain regions in a rat model at progressively more complex levels of feeding-related information processing. The specific aims include: (1) Developing and applying mass defect-based, amine reactive chemical tags coupled with data-independent acquisition (DIA) mass spectrometry (MS)-based strategy for multiplexed quantitation of neuropeptide changes under different feeding conditions; (2) Developing a nanosecond photochemical reaction (nsPCR)-assisted MALDI-based mass spectral imaging (MSI) technique for mapping co-localization patterns of individual isoforms of extended peptide families and amine neurotransmitters in identified neurons and the feeding circuits, with enhanced sensitivity and chemical information; and (3) Assessing functional roles of neuropeptides in feeding and cardiac regulation using a multi-pronged approach integrating in vivo microdialysis and ex vivo physiological and behavioral measurements. Novel neuropeptides will be evaluated for functional roles in feeding regulation at the neuronal network and system levels. The outcome of the proposed research will be a suite of new analytical tools enabling quantitative assessment of the interplay of neuropeptides and biogenic amines with high spatial, chemical and temporal information. The parallel application of these new methods to both crustacean and mammalian nervous systems in feeding will accelerate our pace towards the development of new therapeutics for feeding disorders.

NIH Research Projects · FY 2026 · 2006-02

ABSTRACT Our program to forge principles for how GATA factors act through chromatin to control development, function and dysfunction of the hematopoietic system discovered conserved Gata2 enhancers (+9.5 and -77) that ensure normal hematopoiesis. There were no reports of enhancers essential for stem cell generation or progenitor fate decisions, and few examples of those vital for life. Human +9.5 variants resemble pathogenic GATA2 coding variants in causing GATA2-deficiency syndrome involving immunodeficiency, myelodysplastic syndrome (MDS), and acute myeloid leukemia (AML). -77 disruption also causes MDS/AML. Clinical centers analyze +9.5 and -77 genetic variation. Outcomes from prior research included: (i) innovated a mouse model mimicking epigenetic silencing in GATA2 deficiency syndrome. We demonstrated that GATA2 +9.5 variation is conditionally pathogenic, requiring secondary alterations for pathogenesis; (ii) GATA2 deficiency reduces bone marrow hematopoietic stem/progenitor cell (HSPC) responsiveness to extracellular signals and clinical stem cell expansion regimens; (iii) GATA2-deficient progenitors upregulate innate immune machinery, disrupting cell fate, and GATA2 replacement normalizes the ectopic response; (iv) We demonstrated that GATA2 coding variants can be neomorphic, representing a paradigm-shift; (v) innovated rescue systems with multiomics to quantify GATA2 activity and dissect a novel GATA2 deficiency syndrome variant, which yielded principles for GATA factor function through chromatin; (vi) advised physicians nationally/internationally to interpret GATA2 variation. Pathogenic variants, innovative systems, and multiomics will enable testing of how GATA2-regulated networks govern HSPCs and suppress the onset of blood diseases in which therapies are critically needed. 1. Test models to explain how hematopoietic progenitor genome function is established and maintained. GATA2 activates a Cebpe enhancer to induce C/EBPε expression, promote granulopoiesis, and suppress monocytic differentiation. We will test the hypothesis that GATA2 and C/EBPε operate collectively and independently to establish progenitor genome activity that drives granulopoiesis. 2. Elucidate how a pathogenic GATA2 germline genetic variant retains activity at a subset of GATA2-regulated loci. Our analysis of the ramifications of the GATA2 pathogenic germline variant T354M revealed it retains a subset of its activities. We will test the hypothesis that N-finger activity to counteract the impaired DNA binding of the corrupted C-finger underlies the disproportionate importance of the N-finger for T354M. 3. Generate a human GATA2 genetic curation system to unveil mechanisms and advance clinical genetics. Our assays discriminate activities of human GATA2 variants from wild type GATA2. Although novel GATA2 variants continue to emerge clinically, discriminating between pathogenic, conditionally pathogenic, and benign variants is often inconclusive. We will test the hypothesis that activity metrics will generate a signature for a variant to advance clinical genetic curation, and signatures will also unveil mechanisms.

NIH Research Projects · FY 2026 · 2005-07

SUMMARY The overall goal of our Vision Research Core is to provide expertise, facilities, and equipment to foster and expand vision research at the University of Wisconsin-Madison. This will be accomplished by three Cores. The Ocular Omics and Quantitative Molecular Biology Core will facilitate the use of qPCR, Laser Capture Microdissection, RNASeq, microarrays, NextGen Sequencing, Flow Cytometry and other molecular methods by core users. The Pathology and Imaging Core will facilitate the use of confocal microscopy, fluorescent and light microscopy, histopathology, specialized tissue staining methods, and image analysis by core users. The Animal Models and Eye Organ Culture Core will facilitate the use of non-human primate, feline, rodent and other animal models of ocular disease, the use of OCT, ERG, VEP to assess visual function, and the use of other visual structure and function outcome measures.

NIH Research Projects · FY 2026 · 2004-08

PROJECT SUMMARY / ABSTRACT This project conducts longitudinal Alzheimer’s Disease (AD) biomarker imaging on participants in the Wisconsin Registry for Alzheimer’s Prevention, a midlife observational cohort that is risk-enriched for AD due to parental history of AD dementia. We have been conducting longitudinal amyloid positron emission tomography (PET) with [11C]PiB since 2009 and tau PET with [18F]MK-6240 since 2017 together with advanced MRI, CSF and now longitudinal plasma biomarkers. In the prior funding period we demonstrated that non-demented persons who are Amyloid positive (A+), and tau+ (T+) have been exhibiting cognitive decline since midlife; we developed computational methods to construe amyloid from a temporal standpoint, including individual estimates of A+ onset age and its duration; we found that A+ chronicity predicts cognitive decline and time to dementia—which is ~23 years, giving empirical support to an oft-cited heuristic; we were an early adopter of [18F]MK-6240 for tau PET imaging and delineated its imaging characteristics; we developed and applied cutting edge measures of vascular compromise in the brain with dynamic 4D flow imaging including novel ways for assessing vessel stiffness that is associated with amyloid. The overarching hypothesis of this renewal is that recontextualizing PET amyloid burden along its temporal dimension and anchoring our analyses to A+ onset age and/or its duration will explain heterogeneity in the expression of other biomarkers and cognitive decline. A secondary goal is to determine what can be learned from serial plasma markers of A and T and N (relative to A+ chronology). In Aim 1, anchored to amyloid onset age we will examine factors related to progression of other biomarkers including tau, and examine cognitive decline over a 15 yr period of observation. We will determine the characteristics associated with T+ onset and rate of change (using MK6240 PET) relative to A+ chronicity; we will identify those who are exhibiting disease resistance (i.e., T- in the presence of more chronic A+) and identify the modifiable and genetic factors associated with this biomarker pattern; we will also examine plasma and CSF markers of AD and related disorders. In Aim 2: We will examine conditional processes regarding the added influence of vascular MRI measures on cognitive decline in the presence of progressing AD proteinopathy and patterns of neurodegeneration. Finally, Aim 3 employs computational algorithms to create a joint spatiotemporal profile of amyloid tau and neurodegeneration enabling a joint estimate of disease burden and progression. Significance: This work will fill in the biomarker timeline from late-middle age, providing clarity on the temporal span of preclinical AD proteinopathy anchored to amyloid onset. This project has the potential to greatly improve individualized prognostic precision in view of overall known disease burden, to shed insight on characteristics of disease resistance, and inform prevention strategies.

NIH Research Projects · FY 2025 · 2004-07

Overall Component Project Summary/Abstract This application requests five years of support for the Center for Demography and Ecology (CDE) at the University of Wisconsin–Madison. CDE is a highly-productive population research center, with 70 affiliates in 21 departments across four colleges conducting work that directly addresses the three components of the Population Dynamics Branch scientific mission. CDE has held NICHD center grant funding continuously since 1972, and this application requests continuation of that support under NICHD's Population Dynamics Research Infrastructure Program (P2C). Support is requested for an Administrative Core, a Development Core, and a Scientific and Technical Core. The center grant would support an integrated and interdisciplinary collection of scholars whose research spans the field of population science. During the past five years, CDE has recruited a large number of excellent young scientists and established scholars and strengthened its ties across campus with departments, research centers and institutes in fields related to CDE's research areas. Our research portfolio is now more diverse, more interdisciplinary in character, and covers a greater portion of the life course than in the past. CDE affiliates conduct research in five primary research areas: (1) Fertility, Families and Households; (2) the Demography of Inequality; (3) Health and the Life Course; (4) Biodemography; and (5) Spatial and Environmental Demography. In addition to innovative research in each of these areas, CDE researchers continue to collect and produce high-quality data for use by the population research community, including a growing body of genetic and biomarker data. Continued infrastructure support from NICHD will allow CDE to leverage substantial commitments from the University, a large portfolio of individual research grants, and outstanding human and organizational resources to promote innovative interdisciplinary research in population science.

NIH Research Projects · FY 2024 · 2003-08

ABSTRACT Our community of investigators seeks renewed support for years 15 to 20 of a pre- and post-doctoral training program that addresses the role of microbes in health and disease at the University of Wisconsin-Madison. Microbiology is fundamentally important to human health due to the prevalence and consequence of infectious diseases. Its significance has been elevated by bioterrorism, discovery of the unforseen roles for microbes in causing certain human maladies, and in promoting normal human physiology and health. The proposed Microbes in Health and Disease (MHD) training program represents the natural and synergistic synthesis of the broad disciplines of microbial pathogenesis, beneficial microbiology, and host responses. MHD will have its physical and intellectual home in a state-of-the-art Microbial Sciences Building where basic and clinical scientists interact and collaborate, providing a strong sense of place, cohesion and identity to the Training Program. Our pre-doctoral trainees are drawn chiefly from the Microbiology Doctoral Training Program (MDTP), a top ranked graduate program. Our post-doctoral fellows are drawn from a strong pool of PhD and Infectious Disease MD fellows, the latter from Pediatrics and Internal Medicine programs with a strong history of placing fellows into academic medicine. MHD holds bi-weekly meetings, hosts invited speakers, and has a website and listserve. Medical Microbiology and Immunology, Bacteriology, Medicine and Pediatrics are core departments of MHD and MDTP activities, and offer required didactic, journal club and seminar courses to our trainees. Instruction is provided in host-microbe interactions, microbial pathogenesis, immunology, infectious disease, translational medicine, and responsible conduct of research. Our 35 trainers span 9 departments in 4 colleges and collaborate with each other in research and teaching. All trainers are productive scientists with proven NIH or equivalent funding records and strong records of graduate training. Most are tenured (23 full, 7 associate professors) and 5 promising junior faculty trainers will be mentored by senior training faculty. The training program faculty are dedicated to recruiting outstanding students and fellows, including focused efforts for minority candidates, and are committed to pre- and post-doctoral mentoring and didactic and research training. To support these efforts, and the NIH-stated need to train scientists in the area of microbes in health and disease, support is requested for 8 trainees annually: 5 predoctoral trainees and 3 postdoctoral trainees, including two MD and one PhD fellows. Each trainee is mentored by a committee consisting of a thesis advisor or mentor and 4 other faculty, and all trainees are also co-mentored by virtue of joint trainer service on these committees. Our program and trainers are highly regarded in the scientific community, and fill a unique niche on campus and a critical national need. The success of the program in the prior 5-year cycle is evidenced by 66 publications among the 14 trainees (mean, 4.7 per trainee) that have completed training to date, and their progress into competitive postdoctoral positions or academic, industry or government research careers.

NIH Research Projects · FY 2025 · 2003-07

Abstract The Genomic Sciences Training Program (GSTP) at the University of Wisconsin-Madison is building an inclusive community of leading genomic scientists with strengths spanning across multiple disciplines. The training opportunities and environment we propose will enable our trainees to create and rigorously apply new tools derived from technological advances that are informed by cutting-edge statistical and computational approaches that functionalize diverse and large datasets. The new genomic approaches to biological and medical investigation demand scientists who are knowledgeable and skilled across several fields in effective ways that potentiate new insights or inventions. Accordingly, the emergence of new tools allowing for the creation and interpretation of large-scale experimental efforts is what GSTP has focused on by the didactical interweaving of investigative approaches drawn from multiple fields (biology, genetics, physical sciences, engineering, computer science, and statistics) that were individually contoured for complementing a trainee’s core disciplinary focus, yet built upon achievement and knowledge within the genomic sciences. Given the incredibly rich scientific and engineering breadth of the University of Wisconsin, GSTP has been able to recruit outstanding trainees who greatly advanced mass spectrometry, “omic”-integration, computation, genome engineering, and bio-devices, while exploring new applications leveraging these advantages for cutting-edge investigation into network genomics, proteomics, spatial genomics, metabolomics, and genome biology. These achievements and contributions have nucleated and grown a significant genomics community. This genomics community has become a gateway and central hub for groundbreaking collaborations reaching across departments, centers, schools and other training programs. We propose for the upcoming project period that we continue this focus, with continued emphasis on innovation/invention and fostering of clinical applications, which will advance translational genomics and “turn discoveries into health.” We request funding for 10 predoctoral and 4 postdoctoral traineeships per year. In the next funding period, we will continue to recruit and train trainees who have recently completed their undergraduate or graduate degrees.

NIH Research Projects · FY 2025 · 2002-07

Project Summary/Abstract The University of Wisconsin’s (UW) Computation and Informatics in Biology and Medicine (CIBM) training program is proposing to continue training the next generation of scientists with deep and broad expertise in biomedical informatics and data science. We will continue our collaboration with the Marshfield Clinical Research Institute (MCRI) as a partner in the training grant, and we will enable our trainees to develop their expertise and establish the foundations of their careers within a vibrant ecosystem of biomedical and data science research at UW and MCRI. We will continue our focus on providing trainees with (1) a strong algorithmic and quantitative foundation from computer science and statistics, (2) a broad understanding of the key biomedical informatics and data science methods and challenges, and (3) a solid understanding of the biomedical contexts, spanning the spectrum from molecules to populations of patients, in which methods from informatics can be applied to gain insight and advance human health. Key components of our program include (1) a core set of courses in biomedical informatics and data science, (2) a broad set of supporting electives, (3) a weekly seminar series, (4) an annual retreat, (5) rigorous training in ethics and the responsible conduct of research, (6) rigorous training in methods for ensuring reproducibility, (7) an emphasis on recruiting a diverse pool of trainees, (8) trans-disciplinary co-mentorship, and (9) annual progress meetings with trainees. We have demonstrated strong success in recruiting and training graduate students. This is evidenced by the number of new faculty and other successful researchers we have produced, the development of new externally funded multi-disciplinary research projects, and our track record in underrepresented minority recruitment and placement. We are asking for 10 predoctoral positions for our standard tracks, 2 additional NIAID-supported predoctoral positions for research in biomedical informatics and data science addressing HIV infection, and 4 short-term trainee positions. The CIBM program is well positioned to serve the country with highly trained researchers who have significant expertise and practical experience in biomedical informatics and data science, the foundational disciplines of computer science and statistics, and the biomedical contexts in which these methods can be applied to advance biology and improve human health.

NIH Research Projects · FY 2026 · 2002-03

Project Summary/Abstract In this competitive renewal, we continue our studies of synaptotagmin (syt) isoforms that regulate the release of neurotransmitters and hormones. The founding member of this gene family, syt1, was discovered in 1981 and has been shown to function as the major Ca2+ sensor for synchronous synaptic vesicle (SV) exocytosis in neurons. Surprisingly, the most fundamental property of syt1—it's ability to bind and become activated by Ca2+—remains one of its least understood properties. Syt1 senses Ca2+ via tandem C2 domains, C2A and C2B, and five acidic Ca2+ binding residues have been proposed to coordinate 2-3 Ca2+ ions per domain. However, substitution of these residues in C2A has led to considerable confusion, with conflicting results as to whether these mutations are loss- or gain-of-function or have no effect at all; C2B has been studied in even less detail. In Aim 1 we propose quantitative experiments to compare the roles of each of these putative ligands in binding Ca2+, via isothermal titration calorimetry, and in release, via Ca2+ dose-response measurements of exocytosis using iGluSnFR. In Aim 2, we address the function of syt9, which is perhaps the most misunderstood isoform. Syt9 is in the same clade as syt1 and was proposed to trigger SV release in striatal neurons. Our preliminary data revealed that this is not the case; rather, syt9 is mainly targeted to dense core vesicles in striatal neurons where it regulates the release of substance P, to indirectly control spontaneous SV fusion rates. We will delve into this model by conducting syt9 structure-function studies using both reconstituted fusion assays and syt9 KO neurons, and by conducting pharmacological experiments. We will also expand this work to striatal slices, where substance P has been shown to strongly modulate dopamine release. Importantly, dopaminergic transmission in the striatum plays a key role in addiction and schizophrenia. So, we will conduct experiments to determine whether syt9-regulated substance P release modulates dopamine release in this brain region. Finally, in Aim 3 we return to syt1, to compare its role in activity- dependent SV docking with another isoform, syt7. SV docking dynamics have recently emerged as a key step in numerous aspects of release and short-term plasticity. Using our newly acquired zap-and-freeze instrument, combined with cryo-EM tomography, we will test the idea that syt1 mediates activity-dependent SV docking on short time scales to impact synchronous release, whereas syt7 mediates activity-dependent SV docking on longer time scales to impact asynchronous release, paired-pulse facilitation, and synaptic depression. Collectively, the three Aims proposed here will significantly advance our understanding of the functions of three distinct syt isoforms. In short, we will: 1) address the most elementary questions concerning how the founding member, syt1, senses Ca2+, 2) conduct a rigorous analysis of syt9 function in striatal neurons and, 3) use state-of-the-art approaches to time-resolve the roles of syt1 and syt7 in the newly discovered phenomenon of activity-dependent docking, to—in turn—control elementary aspects of release and plasticity.

NIH Research Projects · FY 2025 · 2001-09

PROJECT SUMMARY/ABSTRACT This application seeks renewal for Years 21-25 of the Training Program in Translational Cardiovascular Science (TPTCS) within the School of Medicine and Public Health Cardiovascular Research Center (CVRC) at the University of Wisconsin-Madison. The CVRC is a campus-wide interdisciplinary research center with faculty and research members from 31 departments in seven schools and colleges, providing a rich environment targeting cardiovascular disease through basic research, clinical investigation, diagnosis and therapy, and public education. The TPTCS training program uniquely creates basic research training for postdoctoral MDs and PhDs (four positions) as well as predoctoral trainees (five positions) with the goal of training scientists who will be capable of thinking and working “from molecule to bedside.” The practical immediate objectives are to attract and train clinicians in basic research, and to attract and train graduate and postdoctoral (PhD) students in clinically motivated basic science. The TPTCS aims to appoint physicians as part of their resident and fellow training to half the postdoctoral positions. One of the long-standing strengths of the program emerging from annual program evaluation is its integration of training among predoctoral students, basic science postdoctoral trainees, and clinical fellows who work side-by-side in journal clubs, in seminar series, and at the bench in laboratories. This blending and sharing of different trainee perspectives is foundational to the goals of the program and creates esprit de corps. The 27 trainers of the TPTCS have been selected from members of the CVRC for the clinical relevance of their science in the focus areas (heart failure/regenerative medicine, arrhythmias/ion channel research, and atherosclerosis/vascular biology), their training records, and the overall robustness and activity of their peer-reviewed research programs. Training takes place at the University of Wisconsin-Madison and exploits the strong institutional support and environment for such training, as well as the established working relationships between clinical and basic science departments and faculty both within the School of Medicine and Public Health and the University at large. Trainers and trainees all participate in core activities that define the program as a cohesive entity. Several innovations are introduced in this renewal, including enhanced clinical integration for the basic science trainees and greater professional development opportunities to prepare all trainees for the next stages in their careers. Appointees to the program to date are extremely strong and program outcomes have been excellent. The scientists and physician-scientists trained in this translational cardiovascular science program are in unique positions to apply basic scientific knowledge to benefit patients with cardiovascular disease, the foremost cause of mortality among Americans.