University Of Colorado Denver

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT For many years, sedation has been integral to managing patients receiving ventilatory support in the intensive care unit (ICU). However, patients who are deeply sedated in the ICU suffer from increased immobilization, delirium, post-traumatic stress disorder, and death. The 2018 Clinical Practice Guidelines for the Prevention and Management of Pain, Agitation/Sedation, Delirium, Immobility, and Sleep Disruption in Adult ICU Patients recommend against the routine use of deep sedation, and the Society of Critical Care Medicine has implemented nationwide campaigns to disseminate evidence-based sedation minimization strategy, the Spontaneous Awakening Trial. Yet, more than 50% of patients who receive mechanical ventilation are being deeply sedated worldwide, and only 20-40% of ICUs regularly practice SAT. Interprofessional teamwork is key to SAT implementation, but optimal strategies to enhance interprofessional team performance around sedation minimization remain unknown. The objectives of this project are 1) to understand how individual ICU staff characteristics and their shared assumptions, values, and beliefs shape their collective approaches to sedation and 2) to promote sedation minimization in the ICU by optimizing the team performance around SAT. This K23 proposal builds on Dr. Fuchita’s preliminary work at the University of Colorado, which reduced deep sedation by promoting a sedation minimization culture. Dr. Fuchita seeks to apply rigorous science to understand the mechanisms of change and develop tools, processes, and strategies that are effective, implementable, scalable, and sustainable across diverse ICU settings. Aim 1 will identify individual ICU staff characteristics associated with intentions to minimize sedation using the Sedation Culture survey. Aim 2 will explore how culture influences interprofessional team dynamics, communications, and collaboration around SAT using qualitative methods. Aim 3 will use Implementation Mapping to develop and pilot PROMISE-ICU (PROmoting MInimal SEdatIon CUlture), a program of multifaceted implementation strategies to improve the adoption and fidelity of SAT at a single ICU and evaluate its acceptability, appropriateness, and feasibility. Dr. Fuchita’s long-term career goal is to catalyze a fundamental culture change in ICUs worldwide to liberate patients receiving mechanical ventilation from unnecessary deep sedation. Dr. Fuchita has co-designed comprehensive career development plans with excellent mentors and advisors to provide him with advanced training in survey and qualitative research, the science of organizational behavior, implementation science, and hybrid effectiveness-implementation trials. The University of Colorado is nationally recognized for its excellence in interdisciplinary health services research, and Dr. Fuchita has unparalleled institutional support and resources to achieve the proposed research, training, and career development objectives. This K23 Award will support Dr. Fuchita’s path toward independence by providing him with the unique skills necessary to design, implement, and evaluate novel interventions to solve complex real-world problems in the ICU.

NIH Research Projects · FY 2025 · 2025-07

Abstract HIV-1 integrase (IN) plays two essential roles in the virus life cycle: i) during early steps of infection, IN catalyzes the covalent insertion of viral complementary DNA into target cell chromatin; ii) during late steps of viral replication, IN binds to the viral RNA genome (vRNA) to yield properly mature, infectious virions. The structural and mechanistic bases for IN catalytic activities have been characterized extensively, and the catalytic activity of IN has been exploited as a therapeutic target. By contrast, the non-catalytic function of IN remains largely underexplored. Initial mutational studies demonstrated that certain IN amino acid substitutions impair proper virion maturation by mislocalazing ribonucleoprotein complexes (RNPs) outside of the protective capsid. This phenotype has been termed class II IN substitutions to delineate them from class I IN mutations that specifically impair the catalytic activity of the viral protein. More recent studies have discovered that IN binds vRNA in virions, and that these interactions are essential for proper virion maturation. IN exhibits distinct preference for select vRNA elements including the trans-activation response (TAR) element. Biochemical assays revealed that IN tetramers rather than dimers or monomers effectively bind to cognate vRNA segments. Furthermore, mutational studies have identified IN residues that selectively impaired IN-vRNA binding without substantially affecting other functions of the protein. The non-catalytic function of IN during virion maturation is an important, yet clinically unexploited, therapeutic target. A new and promising class of HIV-1 inhibitors called allosteric IN inhibitors or ALLINIs potently impair IN-vRNA interactions in WT, but not escape mutant, virus. Consequently, ALLINI treatments yield eccentric non-infectious virions with mislocalized RNPs that phenocopy the class II IN mutations. Despite these remarkable advances, the critical gaps in knowledge remain. Specifically, structural and mechanistic foundations for how IN preferentially binds select vRNA elements and encapsulates the vRNA genome within the mature capsid are unknown. To begin addressing these important questions, we propose the following two specific aims. In Aim 1, we will develop a highly innovative and powerful biochemical model system for assembly of mature virus-like particles (mVLPs), which will examine the ability of recombinant IN to effectively encapsulate vRNA inside capsid-like particles (CLPs). Furthermore, we will test whether class II IN mutations and ALLINI treatments can prevent IN-mediated encapsulation of vRNA within CLPs. In Aim 2, we will determine cryo-EM structures of IN tetramers bound to TAR RNA. Follow up biochemical and virology experiments will validate our structural findings. Collectively, the proposed research will substantially advance our understanding of the non-catalytic function of IN during virion morphogenesis and benefit the ongoing efforts to develop improved ALLINIs for their clinical applications.

NIH Research Projects · FY 2025 · 2025-07

Glycans are a common target of antibody responses. Antibodies against foreign glycans, known as xenoglycans, can protect against highly glycosylated pathogens but can contribute to allergic and inflammatory diseases, such as alpha-gal syndrome, a tickborne disease, and Crohn’s Disease. Therefore, B cells targeting xenoglycans can be harnessed for vaccination and immunotherapy to promote or limit their production of antibodies. However, the B cell subsets and molecular mechanisms underpinning xenoglycan reactivity remain poorly defined. We previously found that humans commonly generate an antibody response against an egg-derived glycan (4S-LacNAc) present in seasonal influenza vaccines. We identified that 4S-LacNAc specific monoclonal antibodies (mAbs) demonstrate some features of natural antibodies produced by B1 cells in mice. However, key distinctions between 4S-LacNAc mAbs and natural antibodies in mice suggest xenoglycan-reactive B cells are a distinct population in humans. Moreover, 4S-LacNAc specific B cell clones could be recalled from year to year, suggesting the establishment of immune memory. To study xenoglycan-reactive B cells, we have generated a blood group antigen A (BGA) specific tetramer. From non-type A human peripheral blood mononuclear cells, we found BGA-specific B cells fell within antigen-experienced B cell subsets. Whether xenoglycan-specific B cells are a distinct B cell subset or are similar to protein-reactive antibodies remains unknown. We hypothesize that xenoglycan-reactive B cells exist within functionally distinct B cell subsets that are primed for antibody secretion relative to protein-reactive B cells and that xenoglycan-specific antibodies utilize restricted repertories, are affinity-matured, and highly specific to a single glycan. First, we will determine if xenoglycan-reactive B cells reside within a distinct memory B cell pool relative to protein-reactive to B cells (Aim 1). Using tetramers for blood group antigens A and B, alpha-gal, tetanus-toxoid, and hemagglutinin, we will perform spectral flow cytometry to determine if there are phenotypic differences between antigen-specificities. Using this information, we will sort these discrete B cell populations to determine if they are enriched for other glycan specificities. We will next determine if these cell populations have increased receptor signaling relative to B cell subsets enriched for protein reactivity. Next, we will define the repertoire and cellular features of B cells targeting common xenoglycans (Aim 2). Using the tetramers described above, we will determine if xenoglycan-reactive and protein-reactive B cells have unique repertoire features and molecular signatures. Next, we will generate mAbs to test if these B cells are undergoing affinity maturation or displaypolyreactivity. Together, the results of this project will provide critical new insight into the nature of glycan-reactive B cells and will inform how these specificities can be targeted by vaccination and immunotherapy, including totreat the tickborne disease alpha-gal syndrome.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT. Despite knowing for more than 60 years that three copies of chromosome 21 (chr21, T21) is the genetic cause of Down syndrome (DS), we still know relatively little about how T21 drives the vast majority of DS pathophysiology. Over the last several decades, alterations in the response to immune signaling have been identified in T21 cells, mouse models of DS and individuals with DS. Indeed, we have shown that the interferon (IFN) response is constitutively active in T21 cells, leading to an interferonopathy-like state of global immune dysregulation, likely due to the presence of four IFN receptors (IFNRs) in a single locus on chr21. Recently, we identified the IFNR locus as a target of directed transient site-specific gene amplification (TSSG) in response to immune activation. Importantly, we found that while the IFNR cluster is subject to TSSG in healthy cells, this process is impaired in T21 cells. Our work to understand rereplication of the IFNR locus has uncovered that innate immune signaling drives rereplication of a network of immune response genes that may be dysregulated in DS. This project will test the hypothesis that innate immune activation drives a heterogenous program of inducible DNA rereplication of immune response loci which is dysregulated by T21. This project addresses Component 1 of the INCLUDE project, targeted high risk – high reward basic science studies in areas highly relevant to DS and aligns closely with the NIAID interest in examination of immune system dysregulation in Down syndrome and its molecular basis. Dysregulation of the immune system and interferon signaling in T21 is a key contributor to many comorbidities associated with DS and our project will delineate further avenues of dysregulation in the immune response in T21 cells. This work will employ a novel single cell amplicon sequencing approach which we leverage to identify transient changes in copy number for the first time. This work will determine how many immune response genes are amplified during immune activation and how this is dysregulated in T21. Furthermore, we will determine if the innate immune driven rereplication program is conserved in primary mouse T cells and if it is dysregulated in a mouse model of DS. Given the novelty of our observation and hypothesis, this proposal seeks to answer a number of questions regarding the innate immune program in T21 including: 1) How do T21 cells suppress TSSG of the immune response? 2) How many immune response loci are amplified in the same cell at the same time and is this heterogeneity different in T21? 3.) Does T21 initiate a different immune response gene amplification program? Using human cell culture models and a novel amplicon sequencing approach we will comprehensively determine the extent of dysregulation of DNA rereplication of the immune response program in T21.

NIH Research Projects · FY 2025 · 2025-07

The extensive impact of firearm injury and death in the United States, involving profound human consequences and economic costs, underscores the need for a comprehensive, evidence-based approach including research and implementation of effective solutions. Historical limitations on funding for firearm-related research contributed to a shortage of qualified investigators, including those from nursing backgrounds, and investment in the next generation of firearm injury scientists is critical. Nurse scientists, with their unique perspective rooted in patient care and a variety of practice settings, play a crucial role in injury and violence research, firearm injury prevention, and overall health outcomes. The University of Colorado is uniquely poised to train these firearm injury prevention scientists from nursing and aligned fields: it boasts resources including a world-class College of Nursing, Firearm Injury Prevention Initiative, and Injury & Violence Prevention Center, as well as a national reputation for expertise and a broad network of partners. Our overarching goal is to address the critical need for a skilled and interdisciplinary workforce capable of researching, implementing, and disseminating evidence-based, innovative strategies to mitigate the impact of firearm-related harms. The Firearm Injury Prevention Research in Nursing (FIPRN) Scholars Program will recruit 8 Scholars on an annual basis for a 12-week program consisting of in-person components, virtual learning, and longitudinal research and career mentorship. Didactic components will provide exposure to key content in firearm injury epidemiology, the evidence base for prevention and intervention approaches, emerging topics, qualitative and quantitative data collection and analysis methods, issues of ethics, innovative methodological approaches, and designing for dissemination. The FIPRN program, grounded in theory but designed for real-world impact, seeks: (1) to provide training in state-of-the-art concepts in firearm injury prevention across settings, populations, and prevention approaches; (2) to provide training in rigorous research methodologies from varied disciplines, attuned to translation into practical solutions; (3) to advance nursing science by developing a workforce of prevention scientists in nursing research who are trained in community-engaged research and bring a range of perspectives, innovative approaches, and non-academic partnerships; and (4) to promote collaborative team-science through active development of interprofessional networks for peer- and senior-mentoring, design and conduct of research, and dissemination of findings. In doing this, we will nurture a group of scientists ready to tackle the complex challenges presented by firearm injury and death prevention and intervention.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Transgenesis to generate reporters or genetic modifiers is a key tool for biomedical research organisms. Zebrafish are a critical model to study early vertebrate development, regeneration, and congenital disease, with transgenic zebrafish as uniquely powerful tools to visualize and perturb biological processes. However, routine generation of transgenic zebrafish using random genomic integration of linearized DNA vectors or Tol2 transposons remains a major challenge in the field: beyond highly variable transgene activity due to chromatin position effects, the required number of animals, resources, and time to select high-quality, single-integration transgenes remain prohibitive to numerous labs. Site-directed transgenesis into so-called safe-harbor landing sites, genomic locations with faithful transgene activity, is critically missing in zebrafish. In Drosophila and mouse, phiC31 integrase-mediated transgenesis of DNA vectors into engineered, carefully selected attP landing sites has revolutionized the field. Access to several attP landing sites enable predictable, reproducible, and economical transgene generation for numerous biomedical research applications, including quantitative assessment of disease variant genes, predictable expression of genetic modifiers, and routine generation of fluorescent reporters. Building on our pioneering work on phiC31 integrase transgenesis, we will establish and expand community-accessible resources for site-directed transgenesis in zebrafish. Replacing two previously established Tol2-based transgenes with attP sites through CRISPR-Cas9, we generated the first possibly universal landing site strains for zebrafish as phiC31 Integrase Genomic Loci Engineered for Transgenesis (pIGLET). We documented reproducible and exceedingly high transgenesis efficiencies upon injection and germline transmission for diverse transgenes. Guided by community feedback to expand on our first two landing sites, in Aim 1 we will generate next-generation landing sites for reporter and enhancer testing applications in our validated loci and will establish previously challenging new Cre/lox tools. However, our current landing sites on chromosomes 14 and 24 limit combinatorial transgene experiments and strain health through inbreeding; additional landing sites with different properties (expression strength, maternal contribution, etc.) on other chromosomes are critically needed akin to Drosophila and mice. In Aim 2, we will streamline mapping of established, faithful transgenes using innovative DNA capture and long-read sequencing to identify suitable additional safe-harbor sites and to generate additional landing site strains and quasi-enhancer traps for diverse biomedical research applications. Our reagents and protocols will be openly accessible through the Zebrafish International Resource Center (ZIRC), Addgene, and direct distribution. Together, our work aims to advance zebrafish transgenesis applicable across NIH institute interests, greatly reducing the required resources, animals, and time to establish transgenic zebrafish strains.

NIH Research Projects · FY 2026 · 2025-07

PROJECT SUMMARY/ABSTRACT Neurodevelopmental (ND) impairment is the most common morbidity for individuals born with critical congenital heart disease (CHD) such as ductal-dependent pulmonary blood flow (DDPBF). This results in high individual, familial, and societal burden. Clinical and sociodemographic factors have been shown to impact ND outcome, and the interventional approach for cardiac repair may also play an important role. For neonates with DDPBF, recent preliminary studies have shown that using transcatheter ductal arterial stenting (DAS) in lieu of surgical systemic-to-pulmonary artery shunting (SPS) may result in improved survival and shorter length of stay but these studies have not examined differences in ND outcomes. These studies led to the COmparison of Methods of Pulmonary blood flow Augmentation in neonates: Shunt versus Stent (COMPASS) Trial, a multicenter randomized controlled trial currently being conducted through the Pediatric Heart Network, that will compare cardiac morbidities and mortality within the first 12 months of life in 300 neonates with DDPBF randomized to SPS or DAS intervention but does not include ND outcomes. The enclosed proposal is an ancillary prospective longitudinal ND study of COMPASS-eligible patients that will compare ND outcomes at 36-months of age between the SPS and DAS groups using a multivariable model to account for pre-procedural medical and sociodemographic risk factors for ND impairment. Candidate biomarkers of neural injury will be measured peri- operatively to evaluate for differences by intervention approach and assess their predictive utility for ND outcomes, while targeted biomarkers of neural development will be measured peri-operatively and at 36-months to determine their relationship to ND outcome. Broader multi-omics analyses will assess for interactions between the genome, proteome, and exposome (including medical exposures and social determinants of health) for predicting 36-month ND outcomes. We will test the following hypotheses: 1) After controlling for known pre- procedural risk factors for ND impairment (medical, sociodemographic, and genetic factors), children who underwent DAS will have better ND outcomes at 36-months of age compared to those who underwent SPS, 2) Perioperative candidate biomarkers of neural injury and longitudinal biomarkers of neural development will improve prediction of ND outcomes in both neonatal intervention groups, and 3). Proteomic, genomic, and exposomic risk factors will interact to further predict 36-month ND outcomes. Results from this proposal will contribute important ND outcome data for the COMPASS Trial and, more broadly, improve our understanding of the ND burden in this patient population through developing the most comprehensive predictive model to date for ND outcomes in children with critical CHD. Our ultimate goal is to identify interventional targets to improve ND outcomes in DDPBF and to inform precision medicine strategies for selecting the neonatal intervention approach based on patient-specific factors to optimize ND outcomes. Finally, this proposal will establish a robust framework for leveraging existing clinical registries to incorporate ND outcomes in future clinical trials in CHD.

NIH Research Projects · FY 2025 · 2025-07

Project Summary This proposal seeks to engage high-school aged youth in establishing schools as clean air environments during and beyond wildfire season. This training and research plan will build upon Dr. D’Evelyn’s expertise in air pollution toxicology and community based participatory research (CBPR) and prepare her for a career in environmental health science and implementation science at the nexus of climate, health, and community action. During the K99 mentored phase, Dr. D’Evelyn will utilize the expertise from her thoughtfully assembled mentorship team and course-based learning to gain training in: 1) best practices in CBPR and Youth Participatory Action Research (YPAR); 2) qualitative methodology, implementation science and evaluation; and 3) exposure science to address health equity questions in a community setting. This training will provide Dr. D’Evelyn with interdisciplinary skills as she moves toward an academic career in independent research. Over the course of the K99 phase, Dr. D’Evelyn will expand air quality knowledge in schools through implementation of an air quality curriculum and a youth-led air quality monitoring network. Aim 1 will establish a permanent monitoring network in the school community which will enable students to collect, analyze and disseminate information about their school’s air quality with the goal of increasing education and awareness of the health impacts of exposure to wildfire smoke. This work will build upon Dr. D’Evelyn’s previous experience working with high school students and will enable her to translate her expertise in air pollution toxicology into an understandable, place-based curriculum to improve awareness and student self-efficacy in smoke-impacted communities. Educational and behavioral outcomes from this aim will be measured through observations and a pre/post survey given to students and teachers to evaluate the impact of their involvement in the project. Upon completion of the K99 phase of this award, Dr. D’Evelyn will move into the R00 phase in which she will complete Aims 2 & 3 that will transition her to an independent researcher. The R00 phase will focus on air quality interventions, implementation science, and evaluation. In Aim 2, Dr. D’Evelyn will continue to foster youth engagement through co-development of an intervention plan that will address the specific exposure concerns of each school community. She will then work with students and school administrators to support implementation of this plan, utilizing the new implementation science skills and knowledge she gained during the training phase. Aim 3 will conclude this independent phase with a full evaluation of the effectiveness of a YPAR project to: 1) reduce exposure to wildfire smoke and other airborne pollutants; and 2) improve community health and health equity in schools impacted by wildfire smoke. This project will describe the impact of a YPAR-based environmental health intervention and implementation program, and work toward a program that could be implemented in any school, regardless of the location or resources available.

NIH Research Projects · FY 2025 · 2025-07

The University of Colorado Anschutz Medical Campus Pharmacology and Molecular Medicine Training Program (PharMM) is currently in its 45th year of NIGMS funding and requests annual support for 8 predoctoral students during the next five years. PharMM distinguishes itself by providing a highly interactive environment in which students obtain a broadly based integrative perspective on science and training in the fundamental knowledge defining pharmacology. The Principal Investigators for the Training Grant are Dr. James Costello, Associate Professor of Pharmacology, and Dr. Mark Dell’Acqua, Vice Chair of the Department of Pharmacology. The Training Program Directors are Dr. Costello and Dr. Jason Aoto, Associate Professor of Pharmacology. Dr. Dell’Acqua is the Chair of the Graduate Training Committee (GTC), which provides the day-to-day oversight for PharMM. The 48 Training Program faculty are drawn from numerous departments across CU Anschutz and have been recruited to provide multidisciplinary training opportunities in neuropharmacology, cell signaling, pharmacogenetics, cancer biology, biomolecular structure, and computational modeling. PharMM faculty are accomplished, committed researchers, and mentors with nearly $900,000 in funding per faculty member. Graduate students are directly admitted to PharMM and recruited from the ‘umbrella’ programs (Biomedical Sciences and Medical Scientist Training Programs). PharMM curriculum includes a comprehensive didactic component, three laboratory rotations, a strong emphasis on student presentations in seminar settings, and a diverse choice of thesis research labs. Career development is well supported by the Graduate School across a range of academic and non-academic career paths. In the last 5 years (2019-2024), PharMM received 148 training grant-eligible applications, with the umbrella programs receiving 1,003 (BSP) and 2,486 (MSTP) applicants. In the past 5 years, 24 training grant-eligible students earned their Ph.D. in PharMM. PharMM has developed and implemented a comprehensive review process to evaluate graduate student applicants, including undergraduate/graduate school grades, research experience and interests, career and research goals, and recommendations from previous mentors and course instructors. Over the past 5 years, the 17 PhD recipients trained by current PharMM faculty have been highly successful – they averaged 2.4 first-author manuscripts and 5.6 total papers, the average time to degree was 5.7 years, ~50% earned individual fellowships, and 100% have a career in science. PharMM currently has 27 TGE (29 total) Ph.D. students. The T32 Training Grant will be critical for PharMM to continue to thrive and meet the national demands for individuals, trained as pharmacologists, who are astute researchers, can be multidisciplinary research team members, and also have the breadth of knowledge to plan and communicate effectively across a spectrum of technologies.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract Type 1 diabetes (T1D) is an autoimmune disorder characterized by destruction of the pancreatic beta cells, leading to decreased production of insulin and hyperglycemia. Although T cells are the primary effectors of beta cell destruction in T1D, autoreactive B cells are also essential to the pathogenesis of T1D. Recent evidence suggests a more important role for B cells in individuals who develop T1D at a younger age and demonstrate rapid progression. However, little is known regarding the specificity, phenotype, and function of B cells in young-onset T1D. Previously we performed a cross-sectional analysis comparing insulin-reactive to tetanus-reactive B cells in the blood of newly-diagnosed T1D and controls using mass cytometry. Unsupervised clustering revealed the existence of a highly activated B cell subset, we term BND2, that falls within the previously defined anergic BND subset. We found a specific increase in the frequency of insulin-reactive BND2 cells in the blood of young-onset T1D donors, which was further enriched in the pancreatic lymph nodes of T1D donors. The frequency of insulin-binding BND2 cells correlated with anti-insulin autoantibody level and phenotypically BND2 cells appear to be capable of potent presentation of autoantigen to T cells. These findings suggest that activation of insulin-reactive anergic B cells likely plays a role in the rapid progression of young-onset T1D. In this proposal we seek to elucidate the mechanism by which these autoreactive B cells become activated and break anergy (aim 1), determine their fate and function, i.e. whether they are, indeed, capable of acting as potent antigen- presenting cells to T cells (aim 2), and determine whether they serve as a biomarker for development of T1D (aim 3). The potential impact of these studies lies in understanding the role of insulin-binding BND2 cells in the rapid progression of disease, which will inform our understanding of the aggressiveness of early-onset T1D and increase the precision of future age appropriate therapeutics.

NIH Research Projects · FY 2025 · 2025-07

Abstract Brain metastasis (BM) confers the worst survival outcomes for patients with breast cancer (BC). While most breast cancer brain metastasis (BCBM) research focuses on triple-negative and HER2+ subtypes, which have higher rates of BM, estrogen receptor positive (ER+) breast cancer, the most frequently diagnosed subtype, accounts for a large fraction of BCBM. Aside from being severely understudied, ER+ BCBM occurs in the unique clinical setting of age and low estrogen (E2). Preliminary data from our lab demonstrates that while models of ER+ BCBM in young mice require supplementation of E2, as is the standard across the field, BCBM were able to outgrow independent of E2 supplementation in older mice, suggesting that the aged brain provides a distinct niche that supports E2-independent outgrowth of ER+ BCBM. Prior studies have identified FGFR1 aberrations as drivers of endocrine resistance and predictors of increased risk for BCBM. Published and preliminary studies show that FGFR1-amplification alone is not sufficient to drive tumor progression in post-menopausal mouse models; rather, FGFR1-kinase signaling activation through the microenvironment promotes tumor progression under low E2. Our preliminary studies show that ER+ patient-derived xenografts (PDX) with FGFR1-amplification have greater ability to colonize the brain, and FGFR1 downregulation decreased the ability of FGFR1+ ER+ BC cells to grow in organotypic brain slices and in the brain in vivo, suggesting that brain-induced activation of FGFR1 may play a role in the promotion of ER+ BCBM. Recent studies have shown that growth of BCBM cells depends on their ability to form pseudo-synapses with glutamatergic neurons, and that neuronal activity can drive BM outgrowth of multiple cancers. Binding of FGFR1 to NCAM (neural cell adhesion molecule) promotes synaptic plasticity and activates FGFR1 downstream signaling in neurons. Our preliminary studies indicate that treatment of ER+ BC with exogenous NCAM induces rapid kinase and downstream oncogenic pathway activation, and that FGFR1 downregulation reduces synapse density in ER+ BC cells co-cultured with brain slices. Thus, the hypothesis of this proposal is that that FGFR1 contributes to ER+ BCBM in the aged and E2- depleted brain microenvironment by i) binding to neuronal NCAM and activating downstream oncogenic signaling and ii) promoting neuronal activity-mediated signaling in ER+ BM. Aim1 will define how interactions between FGFR1 in ER+ BC and NCAM in neurons promote cancer growth downstream of FGFR1. Aim 2 will determine the impact of FGFR1 on the functionality of synapses between ER+ BC and neurons. The long-term goal of this proposal is to study novel interactions between ER+ BC and neurons through FGFR1. This study will identify how FGFR1 in ER+ BC promotes BM outgrowth through neuronal-specific interactions, providing insight into drivers of ER+ BCBM that can inform future therapies and clinical biomarkers.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT Type 1 Diabetes (T1D) is an autoimmune disease where the body’s immune system mistakenly destroys insulin-producing beta cells in the pancreas. While we know that immune cells called CD4 T cells mediate this destruction, we do not fully understand what triggers them to attack. Our research focuses on unique antigens called Hybrid Insulin Peptides (HIPs) that form in insulin-producing cells and may be critical targets of these attacking immune cells. We recently made an important discovery about how these HIPs form. Inside insulin- producing cells are small compartments that normally maintain a specific acid level. We found that in humans, these compartments need to become more acidic than usual for HIPs to form. This happens naturally in mice, which is why scientists can easily study HIPs in mouse diabetes. However, in humans, the compartments may only become acidic enough to form HIPs during times of stress or infection. This discovery led us to propose a new understanding of how T1D develops. Instead of a single triggering event causing continuous destruction of insulin-producing cells, we think the disease progresses in cycles. During infections or other stressful events, the compartments become more acidic, leading to HIP formation. The immune system then attacks cells containing these HIPs. When the infection clears, HIP formation stops, and the destruction temporarily halts until the next triggering event. This explains why some patients develop diabetes quickly while others take years or decades, and why some insulin-producing cells survive even in long-term diabetic patients. Our research will investigate how these compartments become more acidic and how this leads to HIP formation. We will also test whether we can prevent HIP formation by not only controlling the acid levels in these compartments but also by blocking the enzymes responsible for HIP-formation. This could lead to new treatments for T1D that work by preventing HIPs from forming during infections or stress, rather than trying to suppress the entire immune system. Such treatments could help people at risk for developing T1D and protect transplanted insulin-producing cells derived from stem cells from being destroyed by the immune system. This research could transform our understanding of how T1D develops and lead to innovative treatments that prevent the disease by targeting its underlying cause. By focusing on the formation of HIPs rather than broadly suppressing the immune system, we hope to develop more precise and effective therapies with fewer side effects.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Alcohol misuse is a leading cause of liver-related mortality. Incidence and mortality rates of alcohol-associated liver disease (ALD) are increasing, yet there are no effective therapeutics to prevent disease progression and identification of new targets for intervention is essential. Alcohol misuse causes leaky gut. Translocation of gut- derived microbial byproducts [lipopolysaccharide (LPS)] to the liver are important contributors to chronic inflammation and associated liver damage during ALD. Alcohol misuse impacts innate immunity, including complement, and chronic exposure can dysregulate homeostatic immune processes leading to pronounced hepatic inflammation. The resident macrophage of the liver, Kupffer Cells (KCs) are recognized as initiators of ethanol-induced inflammation and key drivers of ALD progression yet the mechanisms underlying KC dysfunction in the liver are unclear. Indeed, ethanol can directly impact KC function by enhancing LPS-induced pro-inflammatory responses (i.e. cytokine IL-1β), leading to hyperactivated phenotypes that perpetuate non- resolving hepatic inflammation. Our preliminary data suggests that ethanol impacts intracellular complement activation (complosome) and increases the localization of a GPCR, complement component 5a receptor 1 (C5aR1) to the mitochondria in KCs. Further, we show that mitochondrial C5aR1 (mtC5aR1) contributes to LPS- and ATP-induced NLRP3 inflammasome activation and IL-1β secretion in vitro. Despite these findings, there remain gaps in our understanding related to the specific role of the complosome as a critical driver of chronic inflammation in the liver. The goal of this proposal is to define novel mechanisms of alcohol-induced metabolic reprogramming of KCs to identify specific, targetable factors amplifying pathogen sensing in ALD. While the complosome has been implicated in cellular metabolism in immune cells, there are no current studies exploring complosome activation in the liver disease of any etiology. It is our working hypothesis that ethanol-induced mtC5aR1 localization is an important mechanism of ethanol priming and hyperactivation in KCs. Specifically, we hypothesize that the C5 complosome mediates ethanol-induced mitochondrial reactive species (mtROS) production and suppression of oxidative phosphorylation, leading to activation of the NLRP3 inflammasome and enhanced IL-1β secretion. Making use of CRISPR-generated C5aR1-/- and C5-/- immortalized mouse KCs (ImKCs) and several C5aR1 antagonists, in Aim 1 we will define if the C5 complosome is required for ethanol-induced mtROS production and immunometabolic reprogramming using in vitro and ex vivo exposure paradigms. In Aim 2, we will determine if KC-mediated liver injury and hepatic NLRP3 inflammasome activation is C5aR1-dependent using a mouse model of ALD and cell-specific C5aR1 deficient mice. In vitro we will elucidate if mtC5aR1 activation is specifically required for KC secretion of IL-1β. Taken together, these studies will reveal whether the complosome coordinates alcohol-mediated chronic inflammation and associated hepatic damage and potentially identify new therapeutic targets to prevent/or treat liver disease.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Bioluminescence imaging (BLI) is widely used for cancer detection and treatment monitoring, but its effectiveness is significantly reduced in hypoxic tumor environments due to the oxygen dependency of traditional luciferase-luciferin reactions. This leads to weak signals and inaccurate imaging, making it difficult to detect and monitor tumors in oxygen-deficient areas. This project aims to overcome these limitations by developing oxygen-independent chitosan-coated gold nanorods (Au-CNRs) that enhance chemiluminescence (CL) imaging precision in hypoxic conditions. These Au- CNRs are designed to trigger a photothermal effect upon near-infrared (NIR) light exposure, releasing singlet oxygen (¹O₂) and producing CL without relying on oxygen. This innovative approach will enable deeper tissue penetration, higher signal-to-noise ratios (SNR), and improved imaging accuracy in both normoxic and hypoxic environments. Key innovations of this project include eliminating the dependency on oxygen and reactive oxygen species (ROS), enabling precise, on-demand ¹O₂ release through NIR irradiation. This results in strong NIR CL at the tumor site, reducing background noise and increasing image specificity and contrast. The Au-CNRs are also biocompatible, stable at room temperature, and minimize toxicity risks compared to conventional inorganic imaging agents. The project will involve synthesizing pyridone-based ¹O₂ release agents with enhanced thermal stability, which will be conjugated to Au-CNRs. These Au-CNRs will be characterized using techniques such as UV-Vis spectroscopy, FTIR, TEM, DLS, and zeta potential measurements. Photothermal studies will evaluate CL intensities, on/off control cycles, and temperature regulation under NIR light to ensure efficient, tumor-specific activation and high-contrast imaging. The expected outcomes include the successful development of thermally stable ¹O₂ release agents and Au- CNRs capable of controlled, NIR-triggered CL in hypoxic tumor environments. This technology has the potential to revolutionize luminescence imaging by enhancing cancer detection and monitoring, particularly in challenging hypoxic conditions, ultimately leading to improved treatment planning and outcomes for patients.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Schizophrenia is a mental health disorder characterized by delusions, hallucinations, disorganized speech, and social dysfunction. It is caused by dysregulated dopamine signaling onto D2-type medium spiny neurons (MSNs). Specifically, dopamine can be released after burst firing of the neuron, which induces phasic release. Phasic dopamine release is elicited during reward prediction as well as in response to salient stimuli, and in schizophrenia, phasic release is aberrantly elicited. This results in the encoding of internal stimuli as salient, leading to the positive symptoms of schizophrenia. Nearly all antipsychotics antagonize the D2 receptor. However, despite their wide use since the 1950s, drug discovery has been halted for the past 70 years. This is because (1) we have limited understanding of how neurons encode phasic dopamine signaling, thereby limiting the pathway specificity and efficacy of newly created drugs, and (2) little is known about how our current antipsychotics change that signaling. Preliminary data in the lab has shown that upon dopamine release and D2 receptor activation, G proteins can engage in membrane-delimited signaling or intracellular signaling with different potencies. However, it is unknown whether these pathways are important for encoding phasic release, and whether clinically effective and effective antipsychotics antagonize either pathway with different potencies. This proposal addresses these two knowledge gaps by defining which postsynaptic pathways are activated by dopamine physiologically, and how clinically effective and ineffective antipsychotics change postsynaptic signaling in a biased manner. In Aim 1, whole cell patch-clamp electrophysiology and 2-photon imaging will be used to define the sensitivity of membrane-delimited and intracellular signaling to dopamine in D2 MSNs. In Aim 2 builds off Aim 1 by examining whether clinically effective and ineffective antipsychotics selectively target these different signaling pathways. Altogether, results from the proposed experiments will allow us to define the pathways necessary for physiologic functioning, how they change in schizophrenia, and as a result which pathways must be targeted for effective treatment. Furthermore, completion of these aims will allow me to gain neuropharmacology expertise, improve my technical skills in electrophysiology and 2-photon imaging, and learn to test rigorous hypotheses applicable to my clinical interests. The training provided and skills enhanced by this proposal will be invaluable for my becoming a psychiatrist with a focus on translating basic neuroscience research into novel therapies for mental health disorders.

NIH Research Projects · FY 2026 · 2025-07

Project Summary The human body hosts a diverse microbial community, interacting through the production, detection, and utilization of a structurally diverse set of specialized metabolites. These metabolites influence community structure by mediating commensal, competitive, and cooperative intra- and inter-species interactions. However, it the system is perturbed, the community can shift from a benign commensal community to a pathogenic one. My group’s long-term goal is to define metabolite-mediated relationships between members of the human microbiota and elucidate their role in community dynamics. We hypothesize that the metabolite-driven interactions of pathogenic communities influence susceptibility of pathogens to antimicrobial treatment. The overall objective of this proposal is integrate cheminformatics analysis of untargeted metabolomics data with physiologically relevant in vitro model systems to elucidate novel chemistry mediating community recalcitrance to antimicrobial therapies. The proposed research is significant because it will provide a fundamental understanding of how specialized metabolites impact the structure and function of the microbiome.

NIH Research Projects · FY 2025 · 2025-07

Project Summary / Abstract Head and neck squamous cell carcinoma (HNSCC) is an aggressive cancer affecting the mucosal linings of the nose and mouth and is associated with smoking, alcohol consumption and HPV infection. Surgery and radiation targeted to the face of cancer patients comes with significant morbidities such as difficulty swallowing, dry mouth, loss of musculature and deformities. The rate of HNSCC is 2.7-times higher in men than women, but is insufficiently attributed to tobacco usage, alcohol consumption or rates of HPV infection suggesting that biological differences between the two sexes contribute to HNSCC. Thus, determining the sex-specific drivers of HNSCC including sex-specific drivers of the immune microenvironment is a critical component to advance precision medicine, matching patients with rational therapies to alleviate patient suffering. Because human patient data demonstrate a critical difference in HNSCC rates in men versus women, the use of animal models of HNSCC will allow us to directly test sex as a biological variable independent of unavoidable, human-related variables. Our preliminary data demonstrate that identical tumors transplanted in male and female immune competent mice grow more quickly in male recipients, tumors in female mice have increased cytotoxic CD8 T cells and monocytes, and tumors in male mice have elevated neutrophils. In our recent publication we demonstrated that neutrophils from tumor-bearing mice possess immunosuppressive activity of myeloid-derived suppressor cells (MDSCs) that inhibit CD8+ T cell activity and promote tumor survival. These data lead us to the hypothesis that sex-specific HNSCC aggressiveness is due, in part, to sex-specific myeloid cell differences in male vs female patients. We will test this hypothesis in the following specific aims: Aim 1: Determine sex-specific myeloid cell differences and activities in transplant models of HNSCC in male and female immune competent mice. Aim 2: Determine sex-specific consequences that deleting PMNs has upon oral tumor progression and the tumor immune microenvironment. Aim 3: Determine the sex-specific myeloid cell microenvironment of human oral cancer specimens. Completing these aims will determine how the immune microenvironment differs in male vs female HNSCCs and the relative contribution of PMNs to tumorigenesis and CD8 T cell activity in males vs females. Establishing these models and collecting these preliminary data will allow us to propose future, mechanistically focused aims in future R01 applications. These studies are critical to our long-term goal to identify cellular and molecular mechanisms that mediate oral malignancy that can then serve as biomarkers and/or targets for personalized therapy to prevent malignancy or more precisely treat patients to improve quality of life.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT: Lymphatic endothelial cells (LECs) act as the first point of contact for antigen and immune cells entering the lymph node (LN) via lymphatic vessels. LECs comprise the subcapsular sinus as well as medullary/cortical structures of the LN. These cells, however, play more than a structural role. Antigen acquired by LECs following protein subunit immunization or viral infection is retained six weeks or more and is actively transferred to dendritic cells for presentation to memory CD8+ T cells. This process of antigen archiving and exchange enhances CD8+ T cell function and local protective immunity. While this process of “antigen archiving” is well supported, little is known regarding the molecular pathways activated at the time of vaccination that facilitate archiving in LECs. Furthermore, the capacity of these cells to retain material from mRNA vaccines has not been evaluated A core requirement for antigen archiving is the presence of an innate stimulus. Adjuvants that induce antigen archiving rely in part or entirely on stimulation of virus-sensing toll-like receptors (TLRs) which lead to rapid type 1 interferon production in the LN. Furthermore, the degree of antigen persistence is dose-dependent on quantity of TLR agonist. Based on our preliminary findings, we hypothesize that protein and RNA delivered by protein subunit or mRNA vaccination persists for longer periods in LECs that have been stimulated by type 1 IFN. Our aims are as follows: (1) Determine if type 1 IFN response in LECs promotes antigen archiving and enhances protective immunity and (2) Determine how type 1 IFN signaling within LECs regulates vaccine RNA stability and downstream protective immunity. These aims are significant because they will lead to an understanding of stimuli that can prolong mRNA vaccine and protein subunit vaccine half-life in the LN and enhance T cell mediated immunity, which is relevant to vaccine adjuvant design.

NIH Research Projects · FY 2025 · 2025-07

Project Summary Concussion is defined as a mild traumatic brain injury induced by biomechanical forces accompanied by an alteration in neurological status. Concussion recovery is complex, and many factors influence short- and long- term recovery outcomes. Among the deleterious effects observed after concussion, sleep-related problems are commonly reported and associated with increased risk of developing persisting post-concussion symptoms. Insufficient sleep is also commonly reported among uninjured adolescents, independent of concussion. In combination, adolescents with concussion are uniquely vulnerable to immediate and persisting sleep deficits. Currently prescribed sleep health recommendations for adolescents with a recent concussion remain vague, with limited evidence supporting intervention guidance. Given the documented relationship between post- concussion sleep problems and poor recovery outcomes, a multidimensional and prescriptive sleep health intervention initiated within the first month of a concussion may lead to improved patient outcomes. Using both patient-reported and objective (actigraphy) methods, our measurement approach will allow us to determine the efficacy of an innovative intervention to improve overall sleep health, concussion recovery, mental health, and academic engagement. The multidimensional sleep health intervention provides targeted and prescriptive recommendations for the patient and is developed from our existing pilot work. Specifically, the intervention addresses five areas with evidence to support post-concussion sleep deficits, including: 1) reduced time asleep, 2) inconsistent sleep/wake time, 3) more bed use for non-night-sleep purposes (e.g., napping), 4) excessive screen time before bed, and 5) night-time anxiety. Participants will be randomized to a multidimensional, prescriptive sleep health intervention or standard-of-care at Visit 1 (pre-intervention), undergo actigraphy monitoring and complete daily surveys for two weeks, return for re-assessment two weeks after Visit 1 (Visit 2, post-intervention), and 8 weeks after Visit 1 (Visit 3, persisting effects evaluation). Therefore, our objective is to discover if a multidimensional sleep health intervention among adolescents with a concussion (1) leads to faster symptom resolution time and better sleep quality and/or duration; and (2) affects persistent sleep habits, mental health, and academic engagement. We will use a two-group randomized clinical trial design (intervention, standard-of-care) with 3 assessments over an 8-week monitoring period. Our multidisciplinary investigative team, with expertise in concussion management, RCTs among adolescents with concussion, clinical trial design, and adolescent sleep behavior and psychology provides necessary experience to successfully complete this study. By challenging current sleep recommendations provided for adolescent patients with concussion, our project seeks to advance rehabilitation strategies for improved concussion management and overall improved health.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Life expectancy for people with cerebral palsy (CP) is increasing – with adults with CP facing a dramatic shift in healthcare access and services upon transitioning to adulthood. Adults with CP have unique healthcare needs and risks beyond physical disability, including higher prevalence of many chronic conditions at younger ages compared to the general population. Many of these chronic conditions are amenable to early detection and treatment through primary care and preventive screening. However, individuals with disabilities including CP face healthcare-related disparities, including lower rates of many preventive services and providers that do not feel comfortable treating patients with disabilities. An online health tool for individuals with Down syndrome improved adherence to healthcare guidelines with high levels of patient and provider satisfaction. Such a tool would require significant adaptation for CP, but would be an ideal tool to adapt given its effectiveness in another common lifespan condition. Dr. Cristina Sarmiento proposes to leverage implementation science frameworks and human-centered design to adapt this online health tool for CP and develop Cerebral Palsy Clinic to You (CPC2U) through the following aims: 1) qualitatively explore design and implementation considerations for CPC2U among primary care providers; 2) adapt an online health tool (CPC2U) to promote appropriate preventive care for adults with CP through human-centered design; and 3) pilot CPC2U to assess acceptability and feasibility. This mentored career development award will support the transition of Cristina Sarmiento, MD, an Assistant Professor of Physical Medicine & Rehabilitation at the University of Colorado, into an independent physician scientist and her long-term career goal of becoming a national leader in lifespan research for individuals with childhood-onset disabilities. To this end, Dr. Sarmiento will complete additional training in these key areas: 1) qualitative methods for intervention design, implementation, and dissemination; 2) human-centered design to develop interventions with community involvement; and 3) designing and conducting pragmatic clinical trials. The University of Colorado Anschutz Medical Campus provides a unique research environment to conduct this research, supported by collaborative research partnerships between the Adult and Child Center for Outcomes Research and Delivery Science (ACCORDS), University of Colorado Hospital, and Children's Hospital Colorado. Dr. Sarmiento's nationally recognized team of mentors, Dr. Megan Morris (disability equity research and qualitative methodology), Dr. James Feinstein (complex primary care research and pragmatic clinical trials), Dr. Edward Hurvitz (lifespan disability and rehabilitation research), Dr. Daniel Matlock (human-centered intervention design and implementation), and Dr. Kathryn Colborn (biostatistics), will guide completion of the proposed training and research plan. This innovative proposal could serve as a model for community involvement in intervention design for other lifespan disabilities.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY The concurrent rise in the prevalence of asthma and obesity poses a significant burden on health care system. Obesity accompanied by loss of insulin sensitivity and compensatory hyperinsulinemia drives severe inflammation and is a defining feature of poorly controlled asthma endotype. While we recognize that obesity exacerbates and promotes asthmatic inflammation, the precise immunological and metabolic mechanisms governing interactions between adipose tissue and the airway remain poorly understood. This knowledge gap presents a significant roadblock in the development of effective therapeutic strategies for obese asthma patients. Our recent novel findings show that in lean mice, adipose tissue recruits eosinophils and undergoes adipocyte beiging and lipolysis in response to airway allergic inflammation in models of asthma. This discovery is of paramount importance as it demonstrates, rather surprisingly, that adipose tissue and its resident eosinophils play essential roles in the systemic response to lung allergic reactions. Moreover, we found that in obese mice, adipose eosinophils prevent obesity-induced airway hyperresponsiveness (AHR) by reducing circulating insulin. Collectively, our innovative findings underscore the vital role of communication between adipose tissue and eosinophils in regulating airway function and inflammation in lean and obese mice. However, the phenotype and regulatory nature of eosinophils in fat tissue, and their role in allergic inflammation with and without obesity, remain unresolved. Based on preliminary evidence, we propose the following central hypothesis: adipose eosinophils mobilize in response to airway inflammation and regulate airway function by influencing adipocyte function and systemic energy metabolism. We will test this hypothesis with the following three Specific Aims: (1) To determine adipose tissue and systemic metabolic response to lung inflammation in lean and obese mice; (2) To determine regulatory nature of adipose eosinophil subset and impact of eosinophils on adipocyte responses during airway inflammation in lean and obese mice; (3) To determine the impact of regulatory eosinophils within adipose tissue on obesity-related lung inflammation and airway hyperresponsiveness (AHR). Completion of these aims will elucidate mechanisms underlying bidirectional communication between lung and adipose tissue in obese and non-obese asthma. Furthermore, this research stands as a pivotal milestone in the development of efficacious therapeutic interventions for managing these interrelated comorbid conditions.

NIH Research Projects · FY 2025 · 2025-06

ABSTRACT The gut microbiome plays a critical role in the pathogenesis of human diseases through their metabolites, acting as a dynamic interface between our environment and health. Recent paired microbiome-metabolome studies enable the simultaneous analysis of hundreds of microbes and metabolites, uncovering their associa- tions with various conditions in unprecedented detail. Integrative analyses of paired microbiome-metabolome data sets require multivariate models that can account for complex correlation structures between microbes and metabolites. However, these multivariate models face the challenge of low statistical power for detecting microbiome-metabolome interactions due to small sample sizes and weak biological signals. Therefore, the overarching goal of the proposed research is to develop novel statistical methods for the powerful detection of weak microbiome-metabolome interactions with limited data. In Aim 1, we will develop a powerful transfer- learning framework for high-dimensional regression models with composition covariates. Unlike most existing transfer-learning methods that borrow information from external data sets, our framework leverages the simi- larities across metabolites within a single cohort. This will help minimize the potential risk of negative transfer, given the different sequencing platforms and bioinformatic pipelines used in different studies. In Aim 2, we apply our transfer-learning method to the human microbiome project (HMP), the integrative human microbiome project (iHMP), and an external paired microbiome-metabolome data set. Our analyses will reveal biologically relevant microbiome-metabolome interactions for both healthy people and patients diagonosed with inflammatory bowel disease (IBD), advancing our understanding of the microbiome-IBD link. Furthermore, we will analyze biopsy- specific microbiome-metabolome interactions for IBD patients, revealing potential spatial heterogeneity for IBD- associated microbiome and metabolites. This proposed research is distinguished by its originality in introducing novel, powerful, and flexible transfer-learning frameworks and novel applications of microbiome-metabolome interactions in both healthy and IBD populations.

NIH Research Projects · FY 2025 · 2025-06

Project summary/abstract We are seeking partial support through this proposal to cover the expenses for the 10th International Meeting of the Middle Ear Mechanics in Research and Otology (MEMRO) 2025 conference, scheduled for June 2025 at the KU Leuven, in Belgium. In alignment with the objectives established at the inaugural MEMRO meeting in 1996, the purpose of this triennial event is to bring together experts in middle ear science and engineering with clinical otologists to facilitate an exchange of knowledge and ideas between these typically independent groups. Previous MEMRO meetings in Shanghai (2018) and Denmark (2015) each attracted approximately 200 participants from over 20 countries. The 2022 meeting had slightly fewer participants since scientists from many countries, including China, Japan, Australia, and others, that would normally attend the MEMRO meetings were still precluded from travel due to COVID restrictions. We expect the 10th edition to have a higher attendance. The 2025 meeting aims to continue promoting the free exchange of ideas between basic and clinical scientists and to establish a framework for innovative collaborative efforts to enhance our understanding of middle ear function, dysfunction, and repair. A primary goal of this meeting is to increase the diversity of attendees, with a particular focus on engaging otology residents and faculty by hosting a hands-on workshop on methods and technologies the day prior to the formal conference. Also many efforts have been made to increase the presence of underrepresented countries. We have promoted the conference on multiple occasions on other world conferences. We have representatives from all continents in the scientific committee. The 2025 MEMRO meeting is being organized by KU Leuven, Belgium's largest and highest-ranked university and is an excellent center for education, research and Innovation. Leuven is a vibrant city only 15 minutes by train from Brussels Airport allowing easy travel, and is rich in art, history, and architecture, offering attendees ample opportunities to experience this dynamic international city.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY/ABSTRACT An important feature of the brain is its ability to combine information about the external environment with internal state in order to generate precisely timed actions. These processes occur on varying timescales and involve different parts of the brain, but the cerebellum is central to shaping sub-second behaviors. Damage to the cerebellum can affect the timing of a conditioned eyeblink and can cause diminished movement endpoint accuracy and oscillatory movements around a target. While considerable work has focused on how cerebellar output can fine-tune movements, it is not known how the cerebellum generates precise temporal representations. This proposal leverages the well-defined structure of the cerebellum and novel optical tools to examine how the synaptic properties of the cerebellar circuit might underlie precisely timed behavioral outputs. Cerebellar granule cells integrate mossy fiber inputs that carry information from different cortical and subcortical streams and that have diverse short-term dynamics. To investigate whether this synaptic diversity generates sub-second temporal representations, genetically encoded calcium, glutamate, and voltage indicators will be used to measure mossy fiber and granule cell activity while mice are performing a skilled reaching behavior that requires temporal precision. Mossy fiber and granule cell activity will be compared to mossy fiber glutamate release to test the hypothesis that mossy fiber activity is transformed at the synaptic level to generate a set of sparse, reliable, and distinct granule cell activity patterns, i.e. a temporal basis set. To test the hypothesis that short-term plasticity dynamics at mossy fiber-granule cell synapses underlie temporal patterns of granule cell activity, a light-activated G-protein-coupled receptor will be used to lower mossy fiber release probabilities. The synaptic consequence of this will be measured using ex vivo whole-cell recordings of granule cells. Further, the effect of altering mossy fiber release probabilities on in vivo granule cell temporal dynamics and on reach endpoint accuracy will be assessed. These findings will elucidate how synaptic diversity can subserve temporal computations in the cerebellum and may provide a more generalizable principle for how synaptic diversity can drive patterns of activity.

NIH Research Projects · FY 2026 · 2025-06

Project Oligodendrocytes nervous propagation. involves proliferation development. developing chemogenetically preliminary precursor development of also maturation. early Here elucidate glutamate Summary (OLs) produce myelin, a lipid rich membrane that wraps neuronal axons in the central system to provide them with metabolic and trophic support and allow for faster action potential Developmental myelination requires precise spatial and temporal regulation that likely communication between OLs and neurons. In the mature brain, neuronal activity promotes OL and differentiation, but less i s known about how oligodendrocytes mature in early brain To test how glutamatergic neuronal activity modulates OL differentiation in the early brain, we use designer receptors exclusively activated by designer drugs (DREADDs) to inhibit or activate activity in cortica pyramidal neurons in mouse pups. Our data show that r educing neuronal activity n early development unexpectedly causes OL cells (OPCs) to differentiate prematurely, whereas increasing neuronal activity in early inhibits OPC differentiation. Here we propose to investigate the long-term consequences early developmental dysregulation of oligodendrocyte differentiation on later development. We will investigate the impact of deprivation of sensory-evoked neuronal activity on oligodendrocyte Additionally, we identified oligodendroglial glutamate receptors as potential regulators of OPC maturation in response to neuronal glutamatergic activity by single cell RNA sequencing. we urther investigate he role of glutamatergic activity on oligodendrocyte differentiation and the mechanisms of glutamatergic signaling to oligodendroglial ionotropic and metabotropic receptors ex vivo. l i f t