University Of Colorado Denver

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Fetal growth restriction (FGR) is the second-leading cause of perinatal morbidity and mortality. Severe cases, especially those with placental vascular changes reflected by absent or reversed umbilical artery Doppler velocimetry (FGRa/r) on ultrasound, are at high risk for perinatal death and exhibit substantially worse outcomes than FGR with preserved fetoplacental blood flow, suggesting that this flow is vital to fetal well-being. The only management option currently is delivery, with guidelines suggesting delivery no later than 34 weeks despite concomitant risks of prematurity. One consistent finding in FGRa/r is inadequate development of placental vessels through the latter half of pregnancy. This sparse vasculature compromises fetal oxygen and nutrient extraction from the maternal intervillous space, giving rise to hypoxia and acidosis, while simultaneously increasing fetal cardiac afterload. Based on a strong correlation between FGRa/r and maternal vascular malperfusion (MVM) – a histopathologic pattern representing placental injury due to defective deep placentation, inadequate uterine vascular remodeling, consequent aberrant intervillous blood flow, and hypoxia-reperfusion (H/R) injury – the conventional view is that FGRa/r is the product H/R injury. However, up to 50% of pregnancies with uncomplicated outcomes also exhibit MVM. Thus, how placental angiogenic deficiencies arise, whether they are solely a consequence of early insults resulting in H/R injury, and how fetal genetic susceptibility to in utero events impacts placental angiogenesis remain major gaps in knowledge. To address these concerns, this R21 application proposes to leverage induced pluripotent stem cell (iPSC) technology and its capacity to abrogate donor transcriptional, epigenetic, and functional signatures. Starting with control (C) and FGRa/r (F) cohorts that display MVM as an indicator of H/R exposure, we will isolate placental stromal fibroblasts (FBs), reprogram them into iPSCs, followed by differentiation into endothelial cells (iPSC- ECs). These iPSC-ECs represent placental ECs naïve to prior in utero exposures while retaining donor genome. By comparing methylomic, transcriptomic, and functional signatures between iPSC-ECs and their subject- matched, primarily isolated FBs and placental ECs, we will be able to rigorously characterize C- and F-iPSC- ECs (Aim #1) and examine the interplay between gene and environment in FGRa/r pathogenesis (Aim #2). Completion of proposed aims will provide critical foundation for (1) identifying targets that promote placental angiogenesis in spite of early in utero insults, thereby increasing surface area for maternal-fetal exchange and improving fetal cardiac function, and (2) generating patient-specific, clinically relevant models with which to test potential therapies.

NIH Research Projects · FY 2025 · 2025-08

ASCENT: OVERALL PROJECT SUMMARY/ABSTRACT Across the lifespan, people living with serious illness suffer physically, psychosocially and financially. The health system too often fails to deliver appropriate, effective care and persons with serious illness (PWSI) frequently experience mismatches between their goals for care and the treatments received. Palliative Care (PC) meets the needs of PWSI and their caregivers by focusing on achieving the best possible quality of life while receiving life-prolonging and curative treatments as is congruent with patient goals of care. PC is appropriate from the point of diagnosis throughout the trajectory of a serious illness for persons of any age, with diagnoses including cancer, Alzheimer's Disease and Alzheimer's Disease Related Dementias (AD/ADRD), advanced organ diseases (including heart, lung, liver, kidney), stroke, neuro-muscular degenerative diseases, genetic disorders and rare congenital and metabolic disorders, among others. While there have been significant gains in the past 2 decades, the evidence base to guide and improve PC – and thus to improve the quality of life for PWSI and their caregivers – remains inadequate. We thus urgently need to expand the PC evidence base and create a robust field capable of creating that evidence base. The Advancing the Science of Palliative Care Research Across the Lifespan (ASCENT) Consortium will be a national consortium of experts in PC that will advance innovative, high-quality research to improve care for people living with serious illness by providing resources and expertise in a coordinated manner. Specifically, the ASCENT Consortium will: 1) Develop the national scientific infrastructure and community needed to advance PC research. The ASCENT community will include scientists, healthcare systems, community-based organizations, clinicians, advocates, patients and caregivers to inform research priorities, study design, and dissemination strategies to maximize the relevance and impact of ASCENT. 2) Create new PC research knowledge and research methodologies by conducting developmental projects, creating a PC research methodology and career development curriculum, and supporting the work of PC scientists via funding and provision of resources and expert consultation. 3) Foster career development and impact of the PC field by supporting career development and pilot awards, providing access to methodologic consultations, and mentoring scientists. 4) Disseminate PC research findings and facilitate subsequent adoption and implementation, employing a multi-pronged approach to dissemination. Through these 4 specific aims, the ASCENT Consortium will transform the field of PC research, generating new knowledge and methods, developing and expanding the palliative care field, and facilitating implementation to enhance the experience of PWSI and their caregivers across the lifespan.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Perinatal hypoxic ischemic encephalopathy (HIE) is globally the second most common cause of death and disability in neonates. Although the brain immune response contributes to the pathophysiology of HIE, no current treatment targets the immune response. A major component of this immune response involves macrophages, which have notable sex differences. Microglia are the brain’s resident macrophages, and they are often grouped together with another type of macrophage found in the brain after injury: peripheral blood monocyte-derived macrophages (MDMs). Despite evidence that microglia may be protective after injury while MDMs may be harmful, many tools do not distinguish between these two types of macrophages. My goal in this grant is to leverage sex to gain a mechanistic understanding of the brain macrophage response after hypoxia-ischemia (HI) in the developing brain. I previously showed that pharmacologically depleting microglia worsens HI-induced lesion in females, but has the opposite effect in males. These data suggest that microglia are protective in female brains; in male brains, microglia may not be as protective, and MDMs may play a larger role after injury. First, I will determine if the primary macrophage contributor to lesion severity after HI is microglia or MDMs, and whether this differs by sex. I will do this using genetic approaches to specifically deplete microglia or MDMs. Second, I will use mice engineered to fluorescently label MDMs to characterize the relative contribution of microglia vs. MDMs after HI in male and female mice. These same mice will then be used for spatial transcriptomics to find the gene expression differences in male and female microglia and MDMs around the HI lesion. Third, I will look upstream of these cells to test if the root of the sex difference in HI lesion after microglial depletion is due to sex chromosomes or gonadal hormones. I will use the Four Core Genotype mice that include XX and XY male mice with testes and XX and XY female mice with ovaries. All together, this work will elucidate the macrophage response to HI in the developing brain. Using sex as a critical variable provides contrast, exposes varying paths to an outcome, and can reveal new therapeutic targets for HIE. My long-term goal is to be a child neurologist who leverages my research expertise in macrophages of the developing brain and clinical skills in neonatal brain injury to develop targeted therapies to improve outcomes for children after HIE. Through this K08 proposal, I will gain experience using transgenic mice, interpreting sex as a critical variable, understanding macrophage biology, and analyzing spatial transcriptomic data. Each member of my mentoring team provides specific expertise to address my training goals. The resulting skill set will leave me uniquely poised as a neonatal neurologist with expertise in the brain macrophage response to perinatal HI. These translational preclinical studies will provide the foundation for my independent investigation and future R01 proposals.

NIH Research Projects · FY 2025 · 2025-08

Perinatal nutritional deficiencies are associated with a higher risk of cognitive and psychiatric disorders, but the cellular and molecular mechanisms explaining this link remain unclear and understudied. Docosahexaenoic acid (DHA), an omega-3 fatty acid found in fatty fish, is an essential building block of brain cell membranes during embryonic development. Importantly, the fetus relies on DHA from the maternal diet to meet the demands of brain growth. Yet, most Americans consume Western-style diets containing very low DHA, raising concerns about the neurodevelopmental consequences for offspring. Low DHA status leads to grey and white matter defects and early-life cognitive impairments. At a cellular and molecular level, perinatal rodent models with DHA deficiency display elevated pro-inflammatory cytokines and activated microglia, the brain’s resident immune cells. Microglia play a key role in the brain’s response to injury and disease. However, during development, microglia also actively sculpt neural circuits through their interactions with neurons and oligodendrocytes. Does DHA deficiency disrupt neuroimmune signaling and critical microglial functions, leading to the observed neurodevelopmental abnormalities? To advance our understanding of how DHA status impacts the developing brain, we will need powerful tools and animal models. Zebrafish are an ideal model system for studying neurodevelopmental mechanisms, as embryos are transparent, enabling direct observation of dynamic cell-cell interactions in the brain at single- cell resolution. To leverage the strengths of the zebrafish model system, I generated DHA-deficient offspring via gene-editing and dietary strategies. With my new model, I am now poised to investigate whether DHA deficiency disrupts neuroimmune gene signatures and cell-cell communication (Aim 1) and the microglia-mediated processes that shape neural circuits (Aim 2). I will also test the extent to which the offspring behavioral phenotype is explained by microglial responses, neuroimmune pathways, and the maternal diet (Aim 3). This project will reveal unparalleled insights into DHA’s role in the brain, potentially informing nutritional strategies that enhance cognitive health. Through my proposed training, I will carve out a research niche at the crossroads of nutrition and developmental neurobiology and lay the foundations for my competitive independent research program.

NIH Research Projects · FY 2025 · 2025-08

The community immediately surrounding the University of Colorado Anschutz Medical Campus (AMC) and the University of Colorado Cancer Center (UCCC) includes underserved secondary schools with a large percentage of students from populations underrepresented in science and research. We aim to expand representation of local students in science careers through an innovative program that fills gaps in cancer research and science education in local schools. Our Students and Teachers Achieving Cancer Research Experiences Together (START) program will inspire 7-12th grade students from underrepresented groups (URs) and their families to see cancer research as an exciting and attainable profession. START will also provide 7-12th grade teachers with research and training experiences to facilitate delivery of novel cancer science curricula in community classrooms. The START program vision is to transform our community by enabling students from URs to envision themselves as the next generation of scientific leaders and pursue advanced biomedical careers while also increasing the local community’s understanding of cancer research and prevention. We will achieve this vision with three aims: Aim 1: Engage secondary students in cancer biology science experiences that expose them to new career pathways while building their engagement and identity in science and research, Aim 2: Transform local science education by engaging science teachers in mentored cancer biology research experiences, providing high quality professional development, and supporting development and delivery of biomedical science curricula, Aim 3: Provide community engagement and outreach opportunities for the neighboring community to learn about cancer research, cancer prevention, and cancer disparities through interactions with AMC scientists. All three aims include curriculum development and implementation components. Outcomes and products of the START program will include novel cancer research curricula, cancer-focused summer camps, college credit for research students, a bioscience lending library for teachers, a teacher network for sharing cancer research teaching ideas and best practices, a teacher certificate with 18 graduate credits, increased interest in research education and careers from local students and families, and community engagement by UCCC centered around cancer research and prevention. To foster students’ continued interest in cancer research, we will provide college preparation sessions and assist START alumni with applying to undergraduate research programs at UCCC. We have a strong pool of scientists to mentor and teach in START, including underrepresented faculty, students, and postdoctoral fellows. START will engage diverse cohorts of 7-12th grade students, teachers, and community members to share in hands-on cancer research and education designed to cultivate ongoing participant success. The proposed aims will address our overall goals by stimulating increased interest in research, generating community and AMC support for local students pursuing research careers, and building the earliest steps in a pathway to biomedical research careers at UCCC.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Single ventricle heart disease (SVD) is a group of potentially lethal congenital heart defects (CHDs) that affects up to 2 in every 10,000 live births in the United States [1]. Children with SVD typically undergo a series of three palliative surgeries, ultimately resulting in the Fontan circulation. Though this staged surgical strategy for treatment has increased short-term survival, experience has shown that the single ventricle is inadequate for long-term circulatory support and patients are hospitalized for many different complications [2, 3], requiring composite outcomes to adequately describe them [4]. We have recently shown that one critical component is the lack of complete understanding of the coupled systemic-pulmonary (Fontan) circulation [5, 6]. Indeed, SVD failure continues to be clinically evaluated predominantly through functional assessment of the single ventricle, with only minor additional consideration given to central venous pressure (CVP), pulmonary vascular resistance (PVR), and to many other comorbidities that often emerge as early as adolescence [7]. This evaluation paradigm also gives little to no consideration of overall vascular compliance, flow pulsatility, or to a host of other heterogeneous disease features, despite emerging recognition that such features have critical importance in overall survival. We hypothesize that parameters of vascular compliance as well as other heterogeneous features independently associate with end-organ damage and will improve prediction of composite outcome. To address this hypothesis, we will noninvasively quantify systemic and pulmonary vascular compliance through novel analysis of previously collected standard-of-care and unique research-based magnetic resonance imaging (MRI) data. With a group of new vascular compliance parameters, together with standard clinical measures, metabolomic, assay, and proteomic measures obtained from a large and highly phenotyped Fontan population, our secondary analysis goal is to develop new prognostic models for SVD patients, and in the long-term implement real time monitoring of physiologic CV parameters that may preemptively identify circulatory failure in children with SVD.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT This proposal presents a revised 2-year research plan focused on improving the understanding of pathologic pulmonary fibrogenesis following acute lung injury and ARDS. The candidate is currently an Assistant Professor of Medicine at the University of Colorado in the Division of Pulmonary Sciences and Critical Care Medicine. The outlined proposal builds on data generated during the candidate’s current NIH/NHLBI K08 award to explore new mechanisms in the pathogenesis and resolution of Fibroproliferative ARDS (FP-ARDS). Data generated from the proposed research strategy will position the candidate for submission of an independent R01 proposal. The acute respiratory distress syndrome (ARDS) is a major healthcare problem in the US. Many ARDS survivors experience more severe disease and impaired long-term outcomes due to the development of pathologic pulmonary fibroproliferation. This excessive fibroproliferation, termed Fibroproliferative ARDS, is characterized by early, over-exuberant fibroproliferative responses with accumulation of myofibroblasts and deposition of extracellular matrix components, due in part to increases in TGF-β. This is followed by a late phase typified by persistent fibrotic changes. There remains a critical need to understand the drivers of FP-ARDS, and to develop therapeutics that attenuate, or reverse, the fibrotic sequelae of ARDS. Oxidant injury and the production of toxic, reactive aldehyde compounds, is key to the pathogenesis of lung injury and fibrogenesis. ALDH2 is a mitochondrial NADP-dependent enzyme that is best known for its role in ethanol metabolism, but is also essential for the detoxification of highly reactive lipid aldehydes. This proposal will evaluate the role of ALDH2 in the pathogenesis of FP-ARDS and test the hypothesis that activation of ALDH2 will promote lung repair and reduce fibroproliferative sequelae of lung injury. The candidate will address two research Aims. Specific Aim 1 will focus on the mechanisms by which ALDH2 inhibits fibrogenesis in lung parenchymal cell (fibroblast and epithelial cell) cultures models. Using genetic (siRNA) inhibition of ALDH2, and specific enzyme activators, we will measure levels of oxidant injury, lipid peroxidation products, and enzyme activity. We will evaluate key steps in the pathogenesis of pulmonary fibrosis, including TGF-- dependent signaling and gene expression in fibroblasts, and senescence phenotypes in epithelial cells. Specific Aim 2 will utilize mice with a genetic deletion of Aldh2 (Aldh2-/-) to determine if absence of this enzyme results in enhanced severity or duration of fibroproliferation in a validated pre-clinical model of FP-ARDS (bleomycin). A pharmacologic activator of ALDH2 will be used in a human Precision Cut Lung Slice model to determine if enhanced enzyme activity can attenuate fibroproliferation, or accelerate resolution and repair. The proposed project will define a novel mechanism in the pathogenesis of fibroproliferative ARDS and explore potential therapeutic targets that could lead to improved pharmacologic interventions for patients at risk for developing FP-ARDS.

NIH Research Projects · FY 2025 · 2025-08

Project Summary: Sickle Cell Disease (SCD) is a genetic mutation that leads to hemoglobin (Hb) polymerization within the red blood cell (RBC), causing the characteristic sickle shape and subsequent hemolysis. Approximately 5-10% of the SCD population suffers from pulmonary hypertension compared to 1 in 3 million in the general population. The 5-year survival rate of SCD patients with PH is ~32%, higher than SCD patients without PH (~16%). A leading hypothesis for why the SCD patient population has such a greater prevalence of PH is that iron toxicity caused from hemoglobin or heme released during RBC hemolysis drives vascular lung injury and inflammation as observed by iron accumulation within the lungs of PH SCD patients. Iron accumulation stimulates an inflammatory cascaded leading to chronic inflammatory conditions within the lung microenvironment. Within chronic inflammatory diseases, such as PH and SCD, a disorganized area of adaptive and innate immune cells amasses in the lung as tertiary lymphoid structures. A critical cell type for adaptive immune cell (B and T cells) activation is the CD169+ macrophage found within the tertiary structure. Classically, these macrophages surround B and T cell areas within the spleen and lymph nodes, providing crucial support and cross talk for adaptive immune system activation. Thus, our overall hypothesize of this proposal is CD169+ macrophages are critical for PH disease progression in SCD. This hypothesis will be tested in three aims. Aim 1 will determine if targeting iron release from the ferroportin transporter modify CD169+ macrophage and adaptive immune cell quantities, phenotypes by multi-omics analysis, and oxidative status. Aim 2 will determine if ablation of CD169+ inhibits PH disease progression by altering the quantity and phenotype of adaptive immune cells within the lung, as determined by flow cytometry, multi-omics approach, and electron paramagnetic resonance. Aim 3 will determine if CD169+ macrophages are critical for adaptive immune cell recruitment to the lung microenvironment. Together, these aims will emphasize the importance of a specific cell type and allow for future projects and therapeutic targets with SCD PH. Receiving this award would drastically improve my career path towards independent research at the University of Colorado Anschutz Medical Campus.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Long before NIDA’s Racial Equity Initiative was launched in the summer of 2020, the Native Children’s Research Exchange (NCRE) was created to be a home for researchers focused on child and adolescent development within Indigenous communities. NCRE conferences provide opportunities for rapid exchange of information and foster relationships that are central to effective research in partnership with communities. Since the first conference in 2008, NCRE has focused on making space for researchers to engage in deep dialogue about their research with others also working to inform better outcomes for Indigenous children through rigorous research. Specifically, NCRE has become a focal meeting for researchers and community partners seeking to grow their understanding of research related to substance use prevention for Indigenous youth. Over the past 16 years, NCRE has fostered a culture of open dialogue and exchange of ideas that has resulted in fruitful cross-institutional and interdisciplinary collaboration. This platform has amplified the reach and impact of the scarce research resources dedicated to Indigenous health research. Many NCRE members are already directly engaged in developing, testing, and implementing prevention programming for Indigenous children and adolescents. Others are engaged in research that has implications for substance use prevention (e.g., research on the impacts of exposure to trauma on children’s developmental outcomes or on the protective role of culture). With a focused Substance Use Prevention Track within the upcoming 2025 NCRE conference, leading researchers and emerging investigators will gather to share their latest work, which can then be leveraged to find innovative strategies to support prevention of early and problematic substance use among Indigenous youth. We will use the NCRE conference platform to implement a Substance Use Prevention Track that will: (1) Build Indigenous leadership in youth substance use prevention research, and (2) Broadly disseminate culturally grounded solutions to Indigenous youth substance use prevention

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Staphylococcus aureus and its methicillin-resistant derivative (MRSA) are the top causative agent of skin and soft tissue infections globally. Though the pathogenic potential of MRSA is unparalleled, approximately 30% of all humans are asymptomatically colonized with S. aureus, mainly in the anterior nares. This colonization by S. aureus is directly correlated to an increased infection probability at multiple sites, making colonization an important step in the S. aureus pathogenic lifestyle. Though S. aureus colonizes a significant portion of the population, a comprehensive understanding of all the factors used by S. aureus to colonize the skin remain unknown. This proposal aims at characterizing the hypothetical protein SasF, and its role in skin MRSA skin colonization and pathogenesis. Previously, our group has performed an RNAseq experiment aimed at assessing MRSA transcriptomic changes that ensue following colonization of healthy murine skin. In this experiment, we observed that sasF was significantly upregulated at both 5- and 24-hours post-colonization. In preliminary studies, we have shown that MRSA SasF is involved in providing MRSA with protection against antimicrobial lipids that are readily found within the skin barrier. We have also shown that a MRSA mutant lacking the capacity to produce SasF is greatly hindered in virulence in a skin-abscess infection model. This proposal seeks to understand how SasF contributes to host skin colonization and infection, and how SasF may mechanistically protect MRSA against these host skin lipids. I hypothesize that SasF protects MRSA against host antimicrobial lipids through charge differences in the wall teichoic acids, and that SasF-mediated cell surface charge modulation provides MRSA with the crucial ability to colonize and infect the skin of a host. These hypotheses will be tested both biochemically and within the context of a host-environments by a variety of in vitro and in vivo biochemical and infection-model approaches in these specific aims: Aim 1: Determine the role of SasF in host colonization and infection and Aim 2: Determine how SasF protects MRSA from skin relevant antimicrobial lipids via the modification of the cell surface charge. In all, the data generated from this proposal will not only increase our collective knowledge of the factors required for S. aureus to colonize and infect skin but will also generate crucial data characterizing how bacteria overcome innate host-lipid defenses. Furthermore, this proposal may help to reveal SasF as a novel antimicrobial therapeutic target which could be used as a target for not only preventing S. aureus skin colonization, but also managing clinically relevant and difficult to treat MRSA infections.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract This proposal is a five-year research plan with an overarching goal of determining the dominant effector functions of granzyme K-expressing CD8 T cells in synovial tissue in rheumatoid arthritis. Inflamed synovial tissue from patients with rheumatoid arthritis contains a large number of CD8 T cells, in some cases representing up to 30% of live cells. We have found that these cells do not express high levels of genes and proteins typically expressed by cytotoxic T lymphocytes, such as GZMB (granzyme B), PRF1 (perforin), and GNLY (granulysin). Instead, these CD8 cells express large amounts of granzyme K, a protease with a cleavage specificity very different from granzyme B. Similar granzyme K-expressing cells have also been detected in tissues in several other autoimmune diseases as well as in many different cancers. Given the abundance of this cell type at the sites of disease in so many different human conditions, determining the effect of granzyme K-expressing CD8 T cells on surrounding cells is vital for understanding their role in disease pathogenesis and for determining how best to target these cells with new treatments. The specific aims proposed here will investigate the effector mechanisms of granzyme K-expressing CD8 T cells using a strategic combination of in vivo, in vitro, and in situ approaches. Aim 1 uses an antigen-induced mouse model of inflammatory arthritis to determine whether granzyme K-expressing cells in inflamed joints are antigen-specific and -activated or whether they are antigen-independent bystander cells, i.e., recruited and activated by the general inflammatory milieu of the synovium. Aim 2 systemically dissects the relative potency of known effector functions of granzyme K-expressing CD8 T cells on the recruitment, activation, and differentiation of myeloid cells, one of the most pro-inflammatory cell types in rheumatoid arthritis synovium and other inflamed tissues. Aim 3 uses multiplex imaging and spatial transcriptomics to identify and compare molecular footprints of granzyme K+ CD8 T cell-associated effector functions in human rheumatoid arthritis synovial tissue samples. This study integrates mouse models, mechanistic cellular immunology studies, and cutting-edge spatial technologies to a newly identified CD8 T cell subset that is highly enriched in rheumatoid arthritis synovium and many other human diseases. This work builds on a long-standing collaboration of the PI, a rheumatologist and translational cellular human and mouse immunologist, and co-I Dr. Fan Zhang, a computational biologist with expertise in synovial tissue -omics analysis and spatial transcriptomic methods development. The findings of this study will inform the design of new therapeutics for rheumatoid arthritis and other autoimmune diseases.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY AND ABSTRACT The University of Colorado Functional Genomics Shared Resource (FGSR) is applying for funds to purchase the Sartorius CellCelector Flex system to bring state-of-the-art live cell and organoid imaging and handling technology to empower cell line development and rare cell detection and analysis. The Major and Minor Users will be 8 investigators, with diverse research interests like oncology, neuro-oncology, immunology, microbiology, biochemistry, and Down Syndrome with 7 active NIH R awards and 5 institutional/foundational grants, and 4 Shared Resources from the University of Colorado Anschutz Medical Campus (CU-AMC). The estimated need for the CellCelector for NIH-funded projects in Year 1 is ~1352 hours (75% of total Accessible User Time or AUT). The requested system will become a part of the FGSR which is a prominent institutional core facility, located within the Department of Pharmacology on the Anschutz Medical Campus and supported by the University of Colorado Cancer Center (UCCC, P30CA046934) and the Linda Crnic Institute for Down Syndrome (Crnic). The FGSR team is composed of a highly qualified research scientist and PI (Dr. Molishree Joshi) and two professional research assistants (PRA). Dr. Joshi has a proven track record of >10 years of providing unrivaled support for functional genomics. The FGSR is the only comprehensive shared resource for functional genomics in Colorado. Their success is reflected by the fact that they have provided a myriad of RNAi, ORF and CRISPR-focused services to >360 research labs in Colorado, supported >280 peer-reviewed publications, and aided in bringing in millions of dollars in federal funds. FGSR was established in 2010 with the mission to support and promote scientific research in Colorado through functional genomics. Since then, FGSR has expanded from a plasmid- distribution core facility to a service center that creates turn-key solutions to support biomedical research. Today, they provide individual reagents for knocking down, over-expressing, and knocking out specific genes and access to carefully amplified pre-made genome-wide and pathway-specific pooled shRNA and CRISPR libraries for genetic screens. They also design and create customized CRISPR libraries and generate model isogenic cell lines with desired genetic modifications. As the core facility progresses towards establishing gene-editing protocols for challenging cells like induced pluripotent stem cells (iPSCs), integrating the CellCelector system for hands-free cell handling is vital. In addition to FGSR’s need for the CellCelector Flex system for the various cell line engineering projects, other users intend to use the platform for 1) cell line development, 2) single cell isolation for multi-omics analysis, 3) single cell cloning of edited cells, 4) picking organoids, spheroids and neurosphere, 5) for selecting rare cell populations from patient samples, 6) single-cell secretion screening and 7) spatial single-cell genomics. Overall, FGSR has a comprehensive technical, financial and administrative infrastructure, generated by strong leadership and institutional support in place, and has rallied a substantial scientific interest supported by NIH funds to support the new system and the FGSR.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Project Summary: This proposal describes a 5-year research training program that will develop Dr. Peter Moore into an independent basic and translational physician-scientist. His long-term career goal is to advance the fields of ARDS, extravascular coagulation, and epithelial repair through the elucidation of mechanisms underlying repair after acute lung injury (ALI). During this K08 Award, Dr. Moore will gain specific career development training and mentorship closely aligned with an innovative research plan. He proposes to study the role of macrophage-produced coagulation factor XIII-A in regulation of the extracellular matrix (ECM) and alveolar epithelial repair after ALI. Given its applicability to multiple pulmonary disorders and repair of injury in other organs, this work is directly relevant to the NHLBI. Candidate: Peter Moore MD, is a board-certified Pulmonary and Critical Care Medicine physician-scientist at the University of Colorado Anschutz Medical Campus. His record of academic excellence, basic science research and scientific publications demonstrates a firm commitment to a career as a clinician-scientist. Training: The proposed career development plan augments Dr. Moore's prior mentored research during his undergraduate, medical, residency, and fellowship training. He proposes to meet his short-term objectives through an integrated combination of intensive mentoring by internationally renowned experts in macrophage biology, ECM proteomics, and transglutaminase activity in tissue repair with didactic and hands-on experiences in (i) extravascular coagulation, (ii) proteomics, (iii) in vivo and ex vivo models of epithelial repair, (iv) scientific writing and presentation, and (v) laboratory leadership. Mentor / Environment: Dr. Moore has close working relationships with highly experienced mentors and collaborators who contribute expertise in macrophage and lung biology (Drs. Janssen, Schwartz, McCubbrey, Mould, Redente), ECM proteomics (Dr. Hansen), extravascular fibrin and transglutaminase activity during tissue injury/repair (Dr. Luyendyk), and innovative tissue imaging techniques (Dr. Schwartz, Redente). The proposed activities will be based at the University of Colorado and National Jewish Health, top-ranked research institutions. Research Project: The primary objective of this proposal is to identify the mechanism by which monocyte- derived macrophage (moMΦ) production of factor XIII-A (FXIII-A) enhances repair after ALI. Specifically, our studies will test the hypothesis that FXIII-A produced by moMΦ at sites of injury crosslinks components to the extracellular matrix that improves type 2 alveolar epithelial cell replication, migration, and differentiation to type 1 alveolar epithelial cells after ALI. This will be tested with well established murine models of acute lung injury and human biospecimens using specialized ECM proteomics, atomic force microscopy, and time-lapsed imaging of precision cut lung slices and decellularized lung scaffolds. In doing so, the specific contributions of moMΦ FXIII-A's modifications of the ECM to repair after acute injury will be elucidated.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Myelin, produced by oligodendrocytes (OLs), is a specialized lipid-rich membrane critical for rapid electrical signal transmission in the central nervous system. OLs are uniquely positioned to modulate circuit activity as they can myelinate up to 40 axons simultaneously, while also differentially distributing myelin on individual axons. Yet how oligodendrocytes select axons for myelination has only recently started to be explored. Our lab has shown that neuronal activity promotes and stabilizes myelin sheath formation and that synaptic vesicle release occurs along axons underlying nascent myelin sheaths. Together, this suggests an intricate mechanism of communication, where oligodendrocytes receive myelination promoting signals from axons. Interestingly, we have found that in OLs, postsynaptic scaffolding and adhesion proteins occupy nascent sheaths and regulate myelin sheath characteristics. Most recently, work from our lab has shown that OLs use specific postsynaptic proteins to myelinate axons based on their neurotransmitter identity. Specifically, we found that the canonical postsynaptic scaffolding protein Gephyrin is enriched in myelin of inhibitory GABAergic and glycinergic axons, functioning to select and regulate the distribution of myelin to these axon classes. If inhibitory postsynaptic proteins guide the myelination of inhibitory axons, do excitatory postsynaptic proteins guide the myelination of excitatory neurons? I hypothesize that the excitatory postsynaptic protein PSD-95 regulates the myelination of glutamatergic neurons by coordinating localization of adhesion molecules. In this proposal, I will utilize genetic manipulation and in vivo and fixed light microscopy in zebrafish to test this hypothesis via two aims. Aim 1 tests if the canonical excitatory scaffolding protein PSD-95 selects and coordinates myelin distribution on glutamatergic axons. Aim 2 tests if PSD-95 regulates myelination by coordinating the localization of excitatory postsynaptic cell adhesion molecules.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Dopamine system is a key neuromodulator that plays multiple roles in regulating locomotion, activity and motor learning within the striatum. As such dysfunctions in dopamine signaling arise in multiple neurological disorders such as Parkinson’s disease. Despite a wealth of literature at the circuit, system and behavioral level it still remains unknown how dopamine signals mediate transmission and how potentially different signals drive different aspects of motor function and learning. To address this the current proposal will use several novel approaches to examine if different dopamine signals exist and if they do what role they may play in gross motor output and motor learning. By altering different aspects of dopamine release and transmission this application will measure local receptor mediated signals as well as quantify bulk spillover dopamine to determine if both aspects are equally perturbed by these manipulations. These different animal models will also be used to see how these perturbations may alter different aspects of gross motor output and motor learning. These experiments will test the central hypothesis that striatal dopamine signals that underlie synaptic vs spillover can be independently regulated and that each regulate different aspects of dopamine’s behavioral functions. The significance of this work will be to determine how dopamine dynamics drive the activation of dopamine receptors to mediate transmission and to determine how those dynamics directly participate in regulating striatal circuits to drive motor activity and motor learning. The proposed studies are expected to be significant in that insights in to the specific mechanisms that regulate dopamine transmission, and are expected to directly lead to testable hypothesis regarding the dysregulations in this system that occur following the loss of dopamine in neurological disorders such as Parkinson’s disease.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY: Trauma remains the leading cause of death for individuals aged 1 to 54, with a significant proportion of these fatalities attributed to trauma-induced coagulopathy (TIC) and a dysregulated inflammatory response, termed thromboinflammation. Advancements in resuscitation have lacked a mechanistic focus, instead, focusing on mitigating the adverse effects of crystalloid fluids, optimizing the use of blood products, and non-selectively addressing clot formation and fibrinolysis. Consequently, improvements in outcomes among trauma patients have plateaued. Further, efforts have failed to address trauma-associated thromboinflammation, which is the primary driver of late-stage morbidity and mortality. To date, there are no therapeutics to target post- trauma thromboinflammation to our inadequate understanding of the mechanistic underpinnings. The protein C system is a key mechanistic driver of TIC and its associated morbidity and mortality following injury and due to its distinct dual anticoagulant and cytoprotective activities, APC both drives TIC development and mitigates thromboinflammation. Thus, therapeutics targeting the protein C (PC) pathway could be ideal for trauma patient management. Supported by our preliminary data, we propose that activation of PC after trauma is an acute protective evolutionary response to preserve cellular function following severe injury, with the unfortunate sequalae of this otherwise beneficial response being TIC. This evolutionary protective response to severe injury is considered maladaptive, providing an early `too much of a good thing' response (cytoprotective AND coagulopathic), which is followed by a late (`too little of a good thing') exhaustion of this response with resultant dysregulated thromboinflammation and organ failure. We propose a dual APC -targeted approach, such that pharmacologic APC-based cytoprotection will help to restore endothelial function and mitigate thromboinflammation of trauma and inhibition of anticoagulant effects will help to curb bleeding and TIC. We are now positioned to explore innovative pathways for mechanistic studies aimed at developing targeted therapies to selectively modulate the protein C system, thereby preventing bleeding and treating thromboinflammation. Here, new activity-selective nanobodies to APC and an engineered factor (F) V variant that bypasses APC anticoagulant activity will be studied to counter the coagulopathy of TIC. Further, engineered 3K3A-APC with minimal anticoagulant activity and full cytoprotective activity, and novel APC-mimetic cytoprotective PAR1 and PAR3 derived agonist peptides will address thromboinflammation. Mechanistic and therapeutic exploration of these novel compounds will address the clinical challenges of TIC and thromboinflammation thereby improving both short and long-term survival among critically injured patients.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Type 1 diabetes (T1D) is an autoimmune disease characterized by the destruction of insulin-producing beta cells in the pancreas, leading to lifelong dependence on insulin therapy. Despite decades of research, the exact triggers that initiate this autoimmune process remain elusive. This grant proposal aims to investigate a novel mechanism that may explain the development and progression of T1D, focusing on the formation and role of hybrid insulin peptides (HIPs) in beta cells. HIPs are unique autoantigens that form when pieces of insulin get linked to other beta cell proteins. These hybrid molecules are not encoded in the genome and may be recognized as foreign by the immune system, potentially triggering an autoimmune response against beta cells. Previous research has shown that HIPs are present in a major mouse model of T1D and human patients, and that immune cells reactive to HIPs can be found in the blood and pancreatic tissue of individuals with T1D. The researchers propose that the formation of HIPs is influenced by various stressors that affect beta cells, such as changes in blood glucose levels or exposure to inflammatory molecules. They hypothesize that these stressors alter the internal environment of beta cells, particularly the acidity levels within insulin-containing granules, leading to increased HIP production. This process may explain why some individuals develop T1D more rapidly than others and why the disease can progress in a variable manner. To test these hypotheses, the research team will conduct a series of experiments using human pancreatic islets and immune cells from T1D patients. They will investigate how different stressors affect HIP formation and examine the relationship between cellular stress, HIP production, and the recognition of beta cells by the immune system. Advanced techniques in mass spectrometry and immunology will be employed to identify new HIPs and characterize their potential role in triggering autoimmunity. Understanding the mechanisms of HIP formation and their role in T1D development could have significant implications for the prediction, prevention, and treatment of the disease. If successful, this research may lead to new strategies for identifying individuals at high risk of developing T1D before symptoms appear, as well as novel approaches to protect beta cells from immune attack. Furthermore, insights gained from this study could potentially inform the development of therapies aimed at preventing or slowing the progression of T1D, ultimately improving the lives of millions affected by this chronic condition. By elucidating the complex interplay between beta cell stress, HIP formation, and autoimmune activation, this research has the potential to fundamentally change our understanding of T1D pathogenesis and open new avenues for intervention in this challenging autoimmune disorder.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The overarching goal of this proposal is to determine the influence of prosthetic foot stiffness on multi- domain loading-related outcomes in individuals with transtibial (TT) bone-anchored limbs (BALs) (i.e., osseointegrated prostheses). BALs are a promising alternative to socket prostheses that have shown to improve overall mobility, physical function, and health-related quality of life. To facilitate positive outcomes, proper load sharing between the implant and bone is pivotal for the mechanobiology of osseointegration at the bone-implant interface, which is directly dependent on loads transferred from the ground to the residual limb. While originated for transfemoral (TF) amputation levels, such implants have recently emerged at the TT level, and one of the primary clinical concerns surrounding TT versus TF BALs is the change in force transmission. While prosthetic foot stiffness plays a direct role in transferring forces from the ground to the implant, there is a paucity of evidence pertaining to how stiffness parameters influence loading at the bone-implant interface and overall clinical outcomes in TT BAL users. As such, current prosthetic prescriptions are largely based on clinical practice guidelines based on evidence originating from socket prosthesis users, which are likely suboptimal. Specifically, due to the difference in force transmission between the two prosthesis types, we identify three significant knowledge gaps that currently influence potential success versus failure/rejection of TT osseointegrated BALs: 1) Standard of care in BAL users is primarily based on evidence from TF amputation levels or socket prosthesis users, 2) Loading is critical to pain and failure rate outcomes in BALs yet cannot be directly measured, 3) Unknown how prosthetic componentry influences force attenuation applied to the implant, sensory feedback, or clinical outcomes. We will address these gaps by implementing a Phase I Clinical Trial that will investigate four Specific Aims: Aim 1) Assess how prosthetic foot stiffness influences bone-implant interface loading in individuals with TT BALs; Aim 2) Determine how prosthetic foot stiffness influences function, pain, and multi-joint biomechanics during activities of daily living in both individuals with TT BALs and standard socket prosthesis users; Aim 3) Establish how prosthetic foot stiffness influences osseoperception and fall risk (balance and reactive control during trip recovery) in both individuals with TT BALs and standard socket prosthesis users; Exploratory Aim 4) Determine the optimal foot stiffness properties that maximize knee joint loading symmetry, minimize stress shielding, and minimize metabolic cost using design optimization coupled with dynamic optimal control methods. By quantifying these relationships, we will provide evidence to support best practices that are specific to TT BALs. In the long-term, these results will impact clinical care by helping develop and guide componentry selection guidelines that are specific to TT BAL users in an effort to facilitate positive outcomes by optimizing bone-implant interface loading in TT BAL users.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Pulmonary hypertension (PH) commonly affects patients with chronic obstructive pulmonary disease (COPD), and associated right ventricular (RV) dysfunction increases exercise limitation, healthcare costs, and mortality. There are no established treatments for PH or RV dysfunction in COPD. Exercise training is a low-risk, cost- effective therapy that improves symptoms and exercise capacity. Exercise training can improve RV dysfunction in some patients, but the lack of personalized, pathophysiology-targeted recommendations is a barrier to effectively using exercise training to improve RV dysfunction in COPD. The proposed K23 training program will address this need by training the candidate in mechanistic clinical trial design and implementation, applying and analyzing multimodal methods to investigate RV dysfunction, and designing and evaluating mechanism- targeted exercise interventions. To support the training plan, the proposed research will investigate patterns of RV response to aerobic exercise and establish the feasibility of an exercise intervention targeted to improve RV contractile response in patients with COPD. Aim 1 is to identify patterns of acute RV response to aerobic exercise. Patients with COPD (n=60) will complete transthoracic echocardiography (TTE) including assessment of RV global longitudinal strain at rest, moderate- and high-intensity exercise. To validate TTE metrics of contractility, a subset (n=16) undergoing clinical right heart catheterization will complete pressure- volume analysis. To identify non-invasive surrogates of RV contractility to apply in future clinical trials, cardiopulmonary exercise testing (CPET) and six-minute walk test (6MWT) metrics will be assessed. Based on the candidate's preliminary data, it is hypothesized that three patterns of RV contractile response will be observed, with one pattern being increase in RV contractility during moderate-intensity exercise but decrease (“contractile exhaustion”) during high-intensity exercise. Aim 2 will assess the feasibility of an exercise training intervention targeted to RV contractile response. Twelve participants from Aim 1 with RV contractility increase during only moderate-intensity exercise will complete a 12-week moderate-intensity interval training program to establish safety and feasibility (adherence). Repeat Aim 1 assessments will assess preliminary efficacy in improving RV contractility. This proposal will lead to multiple peer-reviewed manuscripts and a strong R01 proposal investigating the efficacy of an exercise intervention targeted to RV contractile response to improve RV dysfunction in patients with COPD. The goals of this K23 align with the NHLBI strategic goal of identifying factors that account for individual differences in pathobiology and responses to treatments. The results will tailor exercise training treatments to the individual patient to optimize outcomes. The structured training, expert mentorship, and integrated research aims will prepare the candidate to become an independent physician scientist optimizing exercise prescriptions to prevent and treat RV dysfunction in patients with lung disease.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Alterations in both dopamine (DA) and glutamate signaling in the Nucleus Accumbens (NAc) are thought to play a key role in substance use disorders and addiction. DA regulates NAc circuit activity by modulating glutamatergic transmission and receptor activity, however, the precise mechanisms remain unclear. Synaptic plasticity and regulation of intracellular excitability is primarily attributed to the functions of glutamatergic NMDA recept ors (NMDARs). How dopamine receptors regulate NMDAR activity and how this differs between synaptic connections in accumbal circuitry remain important questions. This proposal will utilize ex vivo slice electrophysiology in combination with optogenetics and iontophoresis in transgenic mouse lines to investigate the synapse specificity and intracellular signaling mechanisms underlying DA receptor regulation of NMDAR current in both D1 and D2 type dopamine receptors. Understanding the synergistic role of D1 and D2 receptor signaling in regulating glutamate transmission and receptor activity in the NAc is a critical step in the investigation of both motivation and the dysfunctions that arise in this system after exposure to drugs of abuse.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT CANDIDATE: Ester Oh, Ph.D., is an instructor, who recently completed her postdoctoral fellowship, in the Division of Renal Diseases and Hypertension at the University of Colorado Anschutz Medical Campus. She has an impressive record of productivity, including prior pre- and post-doctoral grant funding and a strong publication history. Dr. Oh’s ultimate career goal is to independently direct a laboratory focused on evaluating new, evidence- based, pharmacological interventions to improve vascular function and delineate the underlying sex-specific mechanisms in populations at high risk of cardiovascular disease (CVD), including chronic kidney disease (CKD). CAREER DEVELOPMENT PLAN: Dr. Oh’s proposed training plan will address key deficiencies in her training to date and allow her to accomplish her career goal of becoming a successful independent investigator. Dr. Oh’s training plan will consist of: 1) acquiring research skills to conduct fully independent clinical and animal studies; 2) new research skills associated with the proposed research that will distinguish Dr. Oh’s work from those of her mentors and develop her independent line of research; and 3) intellectual and professional skills development through coursework, attendance/presentations at journal clubs, seminars, and national scientific meetings, more grant writing and mentoring experience, and regular interactions with her advisory team. ENVIRONMENT: The environment for Dr. Oh’s training plan will be outstanding. The primary mentor Dr. Nowak (clinical vascular physiology in CKD) and co-mentor Dr. Brunt (translational cardiovascular physiology utilizing both clinical and pre-clinical approach) have been continuously funded by NIH and have strong records of successful mentoring in translational biomedical research. Dr. Oh’s mentorship team also includes two advisory team members Dr. Moreau (sex differences and effects of menopause on cardiovascular physiology) and Dr. Lindsey (G protein-coupled estrogen receptor [GPER] signaling mechanisms). RESEARCH: CVD is a major cause of death in patients with CKD, which is largely attributable to the development of vascular dysfunction primarily vascular endothelial dysfunction and large elastic artery stiffening. In women, there is a greater prevalence of CKD and a faster rate of decline in CKD-associated blood vessel function than in men. However, research on women with CKD is especially lacking. Despite its beneficial effect on the vasculature, long-term use of estrogen-based hormone therapy has raised concerns due to its potential adverse effects. Thus, it is necessary to identify alternative, safe compounds to hormone therapy that can improve vascular function in women with CKD at high CVD risk. One such alternative is equol, a gut-derived metabolite of soy isoflavone daidzein. Equol is a unique phytoestrogen that can activate GPER, which is prevalent in the vascular endothelium and a potential therapeutic target for improving vascular health. The proposed research will test the efficacy of oral equol supplementation for improving vascular function in postmenopausal women with CKD (Aim 1&2A) and examine the mechanistic role of GPER for equol-induced vascular improvements in animal models (Aim 2B).

NIH Research Projects · FY 2025 · 2025-08

Colorectal cancer (CRC) is the second leading cause of cancer death in the US, and differences among population groups are well-documented. A significant proportion of the difference in incidence and mortality between groups is directly attributable to screening rates. Community Health Centers (CHCs), which provide care for millions of patients across the US, have screening rates far below national averages. CHCs are struggling to re-engage patients, at least in part due to staffing shortages and the volume of screenings needed. Recent recommendations to start screening at age 45 has only added to the screening burden for CHCs. Text message reminders are a low cost, scalable way to deliver reminders, especially when integrated into population health management systems that can automatically deploy reminders. These reminders typically have small effect sizes, likely because few text message programs have been optimized to incorporate behavioral theories or communication strategies that can impact effectiveness. We will use our established partnership with the Mass League of Community Health Centers to optimize a mobile messaging intervention available for promotion of CRC screening by fecal immunochemical tests (FIT). With three CHCs from different regions of the US, we will use a two-phase process to first focus on reach, determining optimal messaging for maximizing patient engagement with a text campaign. We will then use a multi-level factorial randomized design to compare strategies for increasing screening completion. The text campaign will be offered with population management tools to track screening and abnormal follow-up completion. We will evaluate the sustainment of the intervention components over 12 months following completion of the intervention period. The proposed work uses a creative, community-engaged, systems-level approach to improve completion of FIT testing and abnormal follow-up among populations served by CHCs. Through the optimization of low burden strategies designed for community health centers, we can create an implementation ready solution to reach more CHC patients with this life- saving screening test. Results will likely be highly generalizable, as CHCs are increasingly turning to population management platforms to support their outreach systems, and thus optimizing their approach to implementing these tools will have widespread benefit.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Down syndrome (DS), caused by the triplication of chromosome 21, is associated with an increased prevalence of autoimmune diseases and neurological dysfunction, potentially driven by chronic inflammation resulting from the overexpression of four interferon receptors (IFNRs). Human chromosome 21 carries four of the six IFNR genes, and their triplication in DS results in significant immune dysregulation and chronic inflammation, characterized by elevated levels of various pro-inflammatory cytokines. This persistent systemic inflammation leads to an autoimmune-prone state, with over 60% of adults with DS experiencing one or more autoimmune diseases. Systemic inflammation and increased cytokine levels may also compromise the integrity of central nervous system (CNS) tight junctions (TJs) — essential structures that form seals between cells in the brain’s protective barriers, preventing harmful substances and immune cells from infiltrating the brain. When these TJs are disrupted, autoantibodies and immune cells can penetrate the CNS, contributing to neurological phenotypes. Despite substantial evidence of CNS TJ dysfunction in DS, including significant correlations between peripheral autoantibodies and neurological phenotypes, as well as elevated levels of a TJ-degrading protein, this critical aspect of neurological health remains underexplored in individuals with DS. This study seeks to uncover the mechanisms by which IFNR triplication induces CNS autoimmunity in DS through a cross-species investigation. By utilizing human samples and mouse models, this project will examine how IFNR triplication contributes to an autoimmune-prone state, CNS TJ dysfunction, and ultimately, CNS autoimmunity. In Aim 1, autoimmune processes affecting the CNS in DS will be identified. Aim 2 will complete a cross-species investigation of CNS TJ integrity in DS. Finally, in Aim 3, mechanisms of CNS autoimmunity will be investigated using a mouse model of DS. The overall hypothesis of this proposal is that systemic inflammation caused by IFNR triplication in DS leads to an autoimmune-prone state and CNS tight junction dysfunction, ultimately resulting in CNS autoimmunity. By elucidating these mechanisms, this study aims to guide therapeutic strategies that stabilize CNS barriers, mitigate immune cell infiltration, and improve neurological outcomes for individuals with DS. Additionally, this research has broader implications, as it sheds light on CNS TJ dysfunction and the effects of dysregulated inflammatory pathways, offering valuable insights into other inflammatory and autoimmune-driven neurological conditions beyond DS.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Obesity is a major risk factor for cardiovascular disease, stroke, and many other adverse health conditions. Weight loss has significant health benefits for individuals with obesity, but long-term weight loss is a considerable challenge for most people, and substantial weight regain is typical within a year of completing a structured weight-loss program. An improved understanding of the drivers of weight loss maintenance is critical to maximize the long-term health benefits. While sustained behavioral change is an important component of weight loss maintenance, the gut microbiome may counteract or exacerbate the effects of returning to pre- dieting lifestyle habits. For instance, gut microbiota characteristics may influence hunger cues and energy expenditure. The goal of this application is to lay the foundation for a comprehensive study of the mechanistic relationships among behaviors, omic profiles, and weight loss maintenance following behavioral interventions. In this project, we collaborate with an ongoing one-year dietary weight loss intervention (R01DK132372; PI: Thomas) comparing the standard-of-care dietary weight loss approach, daily caloric restriction (DCR), to two alternatives, early- or late-day time restricted eating (TRE). We propose to extend the trial by adding a data collection six months after the intervention ends. Our first Aim will compare post-intervention adherence and clinical outcomes (weight, body composition, and markers of cardiometabolic health) between the intervention arms. We hypothesize that TRE will have enhanced adherence that facilitates improved outcomes. Aim 2 will generate pilot gut microbiota data to identify microbial predictors of post-intervention outcomes. We hypothesize that individuals with increases in the abundance of microbial taxa and functions involved in regulating host energy expenditure will have enhanced post-intervention clinical measures, and that these increases counteract the effects of returning to pre-intervention behaviors. This work builds on my ongoing K01 research (K01HL157658), which examines gut microbiota, metabolomic, and genetic pathways during another dietary intervention trial of intermittent fasting versus DCR. My K01 data motivates the hypotheses for this proposed study and will be incorporated into the analyses to enhance the power. This project will provide preliminary insight on how to maximize the long-term health benefits of weight loss strategies and will motivate independent research award applications that combine data from numerous behavioral intervention trials. This work will further my long-term goal of designing more effective clinical interventions that enable both weight loss and weight loss maintenance through innovative, targeted approaches (e.g., offering additional support for individuals predicted to be less responsive, pre-treatment with a microbiome-targeted supplement, or incorporating personalized nutrition therapy). This innovative line of research will complement my ongoing work and advance my career goal to transform obesity prevention and treatment through evidence-based, personalized approaches.

NIH Research Projects · FY 2026 · 2025-07

PROJECT SUMMARY Milk is a complex fluid capable of providing total nutrition of the human infant for 6 months or longer, as well as enhanced protection against infection and common childhood illnesses during the critical early postnatal period. Successful initiation of lactation, called secretory activation, requires hormonal signaling that initiates synthesis and secretion of milk products in secretory mammary epithelial cells and stimulates contractile activity in surrounding myoepithelial cells. Disruptions from maternal obesity, premature birth, or delivery-related stresses are associated with impaired secretory activation and poor lactation performance. Since there are no available treatments for low-milk supply, it is critically important to continue to explore basic molecular mechanisms of mammary gland biology and milk secretion. Specific aims in this proposal explore (1) the role of milk fat secretion in secretory activation, (2) the crosstalk between the secretory cells and myoepithelial cells at secretory activation, and (3) how obesity disrupts milk fat secretion and secretory activation. The proposed studies use mouse models of obesity, quantitative intravital and super-resolution imaging, exploratory and hypothesis-driven omics studies and innovative physiological models to define the cellular mechanisms involved in secretory activation. Genetically modified mice and pharmacological agents will be used to modify specific signaling pathways which may stimulate or inhibit milk secretion. The resulting information will provide a new paradigm for understanding the causes of delayed initiation of lactation in humans, allowing the development of evidence- based strategies to address lactation insufficiency and increase breastfeeding success rates.