University Of Colorado Denver

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT The way antibiotics are delivered to hospitalized children matters. When intravenous (IV) and oral antibiotics are equally effective, the use of oral antibiotics may avoid IV-related harms, improve patient experience, and decrease resource utilization, hospital length of stay, and cost. Pneumonia is the ideal infection to study the overuse of IV antibiotics as it is one of the most common reasons for hospitalization and there is emerging evidence that initial oral antibiotics are likely as effective as IV antibiotics. Despite existing evidence, most children hospitalized with pneumonia still receive IV therapy. This gap in translating evidence into practice highlights the critical need for strategies to increase oral (and reduce IV) antibiotic use. The overall hypothesis is that clinician-focused strategies that target initial antibiotic route in the Emergency Department are feasible, increase oral antibiotic use, and improve patient outcomes. The PI will evaluate these strategies in a pilot hybrid trial that will evaluate both effectiveness and implementation outcomes. Through this, the PI will simultaneously gather data on the effectiveness of IV vs. oral therapy and investigate ways to translate evidence into practice. Specifically, the PI, Jillian Cotter, MD, MSCS, in collaboration with her mentoring team will pursue the following aims: (1) Identify barriers and facilitators to increasing initial oral antibiotics for children hospitalized with pneumonia; (2) Develop a set of implementation strategies to increase initial oral antibiotics; and (3) Conduct a pilot hybrid trial to determine the feasibility and acceptability of the implementation strategies. This will prepare her to submit an R01 grant for a multisite hybrid trial to definitively evaluate implementation and effectiveness outcomes. This proposal focuses on children, an AHRQ priority population, and aligns with AHRQ’s goals to implement studies to accelerate the spread of evidence-based practices. As a pediatric hospital medicine physician and clinical researcher with growing expertise in pneumonia and antibiotic stewardship, Dr. Cotter is uniquely positioned to accomplish the proposed research and training aims. Her career goal is to become an independently funded investigator and national leader in the implementation of antibiotic stewardship practices. Dr. Cotter has developed a detailed career development plan that builds on her research skillset and fills key knowledge gaps that are critical to advancing her career. Through coursework, experiential learning, and guided mentorship, she will gain experience in the: (1) design and conduct of qualitative research, (2) development of evidence-based interventions using implementation science, and (3) conduct and evaluation of pragmatic trials. Dr. Cotter has assembled a strong multidisciplinary mentorship team with expertise in pneumonia, qualitative methods, implementation science, and pragmatic trials, which will ensure her success in achieving her stated aims and training goals. This line of inquiry will prepare Dr. Cotter for future R01 funding and her transition to independence.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT This proposal responds to the National Eye Institute’s (NOT-EY-22-004) call for a better understanding of the connection between social determinants of health (SDOH) and eye health. SDOH refers to “conditions in the environments where people are born, live, learn, work, play, worship, and age that affect a wide range of health, functioning, and quality-of-life outcomes and risks”. Through a systematic review, we identified research gaps regarding SDOH and their relationship with dry eye. Dry eye, a multifactorial disease of the tear film and ocular surface, is a global health problem with considerable impact on quality of life. Dry eye is one of the most common reasons for patients to seek eye care and the societal cost of managing dry eye is substantial. Leveraging multi-center electronic health records (EHRs), we propose to identify indicators of SDOH that are associated with disparities in the diagnosis, management, and prognosis of dry eye. In Aim 1, utilizing a single-center EHR dataset from the Sue Anschutz-Rogers Eye Center, we will explore factors contributing to differential access to care and prognosis among dry eye patients. These potential indicators (or their upstream factors) for SDOH include race and ethnicity, preferred language spoken, and other individual characteristics that are routinely documented in the EHR. We aim to build multivariable prognostic models for generating personalized risk assessment on dry eye progression over a 2-year period, considering important individual and structural barriers to accessing care. In Aim 2, leveraging the multi-center EHR datasets curated by the Sight Outcomes Research Collaborative (SOURCE) Consortium, we will quantify geographical variations in the impact of SDOH on dry eye prognosis. We will validate our initial findings and the prediction models developed in Aim 1 while accounting for additional center-specific factors such as density of eye care professionals. With data from more than 4 million unique patients, the multi-center SOURCE datasets offer substantial sample sizes suitable for external validation. We anticipate providing up-to-date evidence on the association between SDOH and the diagnosis, management, and prognosis of dry eye. By incorporating these routinely collected information as predictors in the rigorously constructed and externally validated prognostic models, we aim to provide physicians with updated evidence for disease management and enhance long-term outcomes and quality of life for dry eye patients. Our findings will inform the development of intervention programs addressing specific factors contributing to disparities in access to dry eye care.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY The levels of fructose are higher in the brains of patients with Alzheimer’s disease, and diets rich in high- fructose corn syrup are associated with an increased risk of developing Alzheimer’s disease. However, the mechanism by which fructose metabolism may accelerate Alzheimer’s disease onset and progression is unknown. Microglia—the innate immune cells of the brain—protect against or promote the development of AD, depending on their function. Notably, microglial function is regulated by metabolism. When activated, microglia shift towards glucose utilization and away from oxidative phosphorylation. Chronic activation eventually leads to ‘metabolic reprogramming’ and microglial dysfunction, which drives AD pathology. Although strategies targeting microglia hold enormous potential for treating AD, molecular targets that can help reverse the metabolic programming of microglia have not yet been identified. The fructose transporter GLUT5 (also known as solute carrier family 2-member 5 [Slc2A5]) is predominantly expressed by microglia of the mammalian brain and has recently been identified as one of several genes within a ‘microglia-specific’ signature linked to late- onset-AD pathology. Our compelling preliminary data has also shown that GLUT5 is increased in the brains of aged and 5xFAD mice and that down-regulating fructose metabolism improves age-associated changes in microglial morphology. Therefore, we hypothesize that GLUT5-mediated fructose metabolism may drive the metabolic reprogramming of microglia associated with AD pathology. However, due to the lack of available genetic tools, the role of fructose metabolism and microglial GLUT5 in AD pathology has not been empirically determined. This R21 proposal brings together a scientific team uniquely positioned to interrogate this critical question: Dr. Bruce (microglia, metabolism, neurodegenerative disease); Dr. Lanaspa (fructose metabolism, cardiometabolic disease); Dr. D’Alessandro (metabolomics): and Dr. Frietze (bioinformatics, single-cell omics). In Aim 1, we will characterize the first conditional GLUT5flox/flox mice and generate the first microglia-specific GLUT5 KO mouse to empirically determine the role of GLUT5 in microglial metabolism and function. In Aim 2, we will utilize AD susceptible transgenic mice and mice lacking GLUT5 and Ketohexokinase (KKH, rate-limiting enzyme in future kinase) to determine whether limiting fructose metabolism prevents metabolic programming and microglial function in AD. In Aim 3, we will use human microglia-like cells derived from peripheral blood mononucleocytes to test whether pharmacologically inhibiting GLUT5 can restore age- associated dysfunction in human microglia. The impact of this study is threefold: 1. To develop novel genetic tools of great interest to AD researchers and beyond; 2. To understand the mechanisms driving fructose- mediated metabolic reprogramming; 3. To test interventions that limit fructose metabolism, validating GLUT5 as a target to improve microglia function in AD.

NIH Research Projects · FY 2025 · 2024-09

There has been a monumental shift in the breast/chestfeeding (B/CF) recommendations for mothers and other parents with HIV (MoPWH) in the United States (US) from complete avoidance to a shared decision-making approach. For decades following the start of the HIV epidemic, US guidelines recommended against B/CF for MoPWH to avoid any risk of HIV transmission to infants. However, current US guidelines now recommend an evidence-based, shared decision-making approach to infant feeding counseling between providers and MoPWH who desire to B/CF. Yet, there is very limited evidence on infant feeding decision-making, practices, and outcomes among MoPWH in the US to inform best practices of this shared decision-making approach. Data are unavailable to identify which demographic, psychosocial, and clinical factors influence infant feeding intention, practices, and outcomes. Further, there is a need to better understand current institutional practices regarding infant feeding and the perspectives of the multidisciplinary teams who care for MoPWH and their infants. To best support shared decision-making while minimizing HIV transmission, there is an urgent need to collect representative epidemiologic data from B/CF MoPWH in the US as well as comprehensive insight from MoPWH and other key stakeholders on their perspectives and experiences. The proposed UPLIFT Study (Understanding Parental Lactation and Infant Feeding decisions Tailored to people with HIV) will address the knowledge gaps outlined above in response to the Centers for Disease Control and Prevention Request for Applications RFA-PS-24-040: Understanding Infant Feeding Preferences, Practices, and Outcomes for Mothers and other Parents with HIV in the United States. In collaboration with the International Maternal Pediatric Adolescent AIDS Clinical Trials (IMPAACT) Network and by leveraging IMPAACT’s US sites, we will use a convergent mixed methods research design to accomplish three overarching core activities. In Core Activity 1, we will explore the multifaceted process of infant feeding decision-making via in-depth interviews with key stakeholders, including a diverse sample of MoPWH, their partners and other influential individuals, and care providers. In Core Activity 2, we will establish an observational longitudinal cohort of N=450 MoPWH at 10-15 representative sites across the US to examine current infant feeding practices and outcomes. We will characterize current standards of practice for infant feeding in the US using a landscape analysis; assess factors associated with infant feeding decisions and outcomes; carry out human milk HIV viral laboratory assays; conduct exit interviews and mixed methods integration; and perform costing analysis. Finally, in Core Activity 3, we will develop and pilot a national voluntary registry to capture data from the diverse population of MoPWH who B/CF in the US. Findings from the three proposed core activities in the UPLIFT study will inform best practices for infant feeding counseling to promote equitable patient-centered care for MoPWH in the US.

NIH Research Projects · FY 2024 · 2024-09

Abstract The aging of the eyes is a phenomenon that many of us take as a given as we age, vision loss and the subsequent impacts on daily life - less physical activity, higher risk of depression, higher risk of falls, more rapid cognitive function decline - considered immutable consequences of the aging process. Age-related vision loss results from myriad pathways but systemic inflammation and oxidative stress may be a common underlying factor. Not everyone will develop eye disease, though most experience vision changes with age. The vital question is: are there ways to distinguish between individuals along the continuum from normal age-related changes to faster deterioration of vision function to overt pathology? Having an indicator of underlying pathological processes that contribute to faster aging of the eyes would have a large impact on screening and vision care as well as identify high risk groups for clinical trials. Mitochondria, which are central to the production of adenosine triphosphate (ATP), to the regulation of bioenergetic processes, and to various signaling pathways, are susceptible to oxidative stress, resulting in loss of efficiency in oxidative phosphorylation. Mitochondrial damage is a major contributor to age-related diseases through cell apoptosis, dysregulated energy production, and inflammation. Mitochondrial function is increasingly thought to be an important indicator of systemic aging, and mitochondrial diseases frequently involve a range of ophthalmic manifestations. We hypothesize that mitochondrial function and genetics may provide a marker of the biological aging of the eyes as well as provide information on susceptibility to pathologic damage leading to disease. Through this project, we will identify the role of mitochondrial function and genetics in the etiology of aging-related eye diseases. We will build a platform and foster existing collaboration to further investigate the role of mitochondrial function in age-related eye diseases. Lastly, this project will generate knowledge for the development of both mitochondrial function-based clinical indicators of age-related vision loss and provide an evidentiary base for potential interventions on modifiable aspects of mitochondrial function that could slow the aging of the eyes to improve vision health.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Despite an increased investment in primary care screenings to identify the psychosocial needs of caregivers and children (e.g., exposure to violence, racism, and insecure housing), these efforts don’t always result in family-centered service referrals and follow-up to ensure all needs are getting met (i.e., families able to access services such as housing and behavioral health support). When psychosocial needs aren’t identified or addressed during childhood, it can cause or worsen children’s health conditions, interrupt their development, and, at the societal level, perpetuate disparities in overall health. Our preliminary data identified key strengths and weaknesses within families’ journeys through psychosocial screenings, service referral and linkage processes in primary care – which we term “service pathways”. The goal of the current study is to optimize the post-screening pathway to increase family referral and linkage follow-up, so that all screened families receive consistent services. We hypothesize that clinics using our proposed adaptive intervention package will increase family referral and linkage follow-up compared to clinics using current screening practices. This study will scale up preliminary work in a large safety net healthcare system to optimize referral-to- linkage service pathways. Process Service Mapping (PSM) is a novel and generalizable methodology that actively engages partners to better understand how processes actually work, and ways to optimize service pathways. PSM maps patients’ journeys using iterative cycles to identify inequities, challenges, and action points. This study will scale up previous work by testing the CARELOOP Intervention (Clinics cAtch needs, REfer, Link to services, and close the lOOp using an equitable family-centered Process). CARELOOP is a system-level intervention designed to enhance psychosocial screenings through PSM methodology and Implementation Science, and grounded in the Clinical-Community Relationships Evaluation framework. Denver Health is a large safety net system serving minoritized families and 11 pediatric clinics already screen for psychosocial needs using the Health-Related Social Needs and Survey of Well-being of Young Children. Our main hypothesis is that intervention clinics implementing CARELOOP will have higher Service Referrals and Linkages compared to standard care control clinics. After optimizing service pathways (Aim 1), we will conduct a cluster-randomized trial to test the impact of CARELOOP on effectiveness (Aim 2) and implementation outcomes (Aim 3). The study goals are: Aim 1: Engage clinics and communities to refine CARELOOP by mapping service pathways to include equity and family-centered elements and tailored strategies; Aim 2: Use a parallel-arm cluster randomized trial to test the effectiveness of the CARELOOP intervention; and Aim 3: Conduct an implementation evaluation. This R01 study leverages current psychosocial screenings to rigorously test an intervention designed to increase service referrals and linkages. We bring quality improvement, implementation science, and novel co-creation engagement to accelerate family-centered care.

NIH Research Projects · FY 2024 · 2024-09

Growing evidence suggests the contribution of altered brain microcirculation to cognitive impairment and dementia observed in Alzheimer's disease (AD) and AD-related dementia (ADRD). Yet, the lack of approaches to image and investigate the function of the small cerebrovasculature has hampered our progress in understanding the pathological sequence of vascular cognitive impairment and dementia (VCID). The earliest signs of AD and VCID in patients and mouse models typically involve deficits in cerebral blood flow. Specifically, neurons lack energy reserves and thus rely on a “just-in-time” neurovascular coupling (NVC) strategy in which active regions signal to the microvasculature to locally dilate and increase local blood flow. Patients and mouse models of AD or CADASIL, a monogenic archetypal form of VCID, show an early deterioration in NVC. Our previous studies have identified a molecular defect at play in capillary endothelial cells: depletion of phospholipid PIP2 prevents Kir2.1 channels to act as sensors of increases in external K+ — a product of neuronal activity — and transduce this into a vasodilator electrical signal that rapidly propagates to upstream arterioles, driving vasodilation and local hyperemia. We further linked PIP2 depletion to a lower ATP/ADP ratio (i.e., phosphorylation potential) in CADASIL capillary endothelium. Our multidisciplinary team, with complementary expertise in cutting-edge imaging of brain microcirculation and bioenergetic approaches, will test the hypothesis that alterations in the extracellular matrix inhibit autocrine activation of the epidermal growth factor receptor (EGFR) leading to mitochondrial dysfunction and lower ATP production in CADASIL but also in presence on Aβ oligomers. We further propose to investigate this pathomechanism in humans using freshly isolated brain microvessels paired with spatial and single cell transcriptomics in human autopsy brain tissue. To attain this goal, we will engage a wide variety of novel, state-of-the-art experimental approaches using intact animals, native tissue, and freshly isolated cells, complemented by sophisticated multi-omics analysis. Aim 1 will investigate Aβ- induced metabolic impairment in capillary endothelial cells in cerebral amyloid angiopathy (CAA) and Alzheimer's disease (AD) over time. Using extracellular matrix disruptions characteristic of CADASIL as a framework, Aim 2 will elucidate the mechanism by which EGFR inhibition impairs mitochondrial function in capillary endothelium. Finally, Aim3 will create an integrated view of the pathological role of EGFR inhibition and endothelial energetic impairment in in AD/ADRD by investigating this pathway in CAA and AD. The proposed work has the potential to provide a paradigm-shifting view on how capillary endothelial cell energetics control neurovascular coupling, and as such, should provide the foundation for understanding VCID development that is necessary to identify novel efficacious therapeutic strategies.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Proteins are the molecules that carry out the majority of biological function. Although mRNA levels can be measured at scale and have been transformative for understanding gene expression in large cohorts, mRNA levels correlate only partially with protein levels in a system. As a result, some differentially expressed genes from transcriptomics experiments may not be informative for the abundance of their proteins, leaving their functional significance difficult to interpret and leading to a loss of information that impedes the translation of `omics' experiments to biological knowledge. The recent availability of large matching transcriptomics and proteomics data has created new avenues to predict protein level changes from mRNA profiles using machine learning methods. Results from these efforts have highlighted the prevalence of post-transcriptional regulation of the proteome, where the abundance of a protein species in a sample is often determined not only by its own coding mRNA, but the abundance of other mRNAs in the transcriptomes, including many of those coding for its protein-protein interaction partners. Accordingly, this project aims to explore new strategies that capture protein-protein relationships to enhance our current capability to infer protein-level changes from mRNA abundance measurements. Specifically, Aim 1 will explore the use of conceptual embeddings of proteins to create low-dimension vectors that capture relevant protein information on: (1) the topology of protein-protein interaction network measured in large mass spectrometry experiments, and (2) protein sequence, domain, and structure information; and then evaluate their utility for capturing the relevant protein neighborhoods that aid in the prediction of proteomic changes from mRNA abundance. In parallel, Aim 2 will aim to disseminate technological advances by building enabling software tools and web apps that will take the pre-trained models to analyze new user input mRNA sequencing results, which are designed to assist in the prioritization and interpretation of gene lists from sequencing experiments. The models will be validated by mass spectrometry and immunoblot experiments. If successful, the proposed work will lead to broadly applicable software tools that can enhance the utility and interpretation of transcriptomics and proteomics experiments. It may also yield new insights into the biological factors that contribute to non-correlation between mRNA and proteins.

NIH Research Projects · FY 2025 · 2024-09

Project Summary: Vascular dementia is the second most common cause of dementia. Therapies aim to control risk factors (e.g. hypertension, hyperlipidemia, and diabetes mellitus) to prevent development or worsening of disease. Recent studies show that viral infections are potentially modifiable risk factors for dementia. Amongst all pathogens, varicella zoster virus (VZV) is the most likely contributor to vascular dementia. VZV is a neurotropic DNA virus that is latent in >95% of Americans and reactivates to produce herpes zoster (shingles) in 50% by 85 years of age. Despite the availability of zoster vaccines, there are still >1 mil cases of zoster annually. Recent studies reveal zoster increases dementia risk and treatment (vaccines, antivirals) reduces risk. Importantly, a recent FinnGen and U.K. biobank study showed VZV was specifically associated with an increased risk of vascular dementia. VZV likely contributes to vascular dementia risk through its well-established ability to produce vascular pathology leading to ischemic or hemorrhagic stroke (VZV vasculopathy). VZV directly infects cerebral arteries and causes a vasculitis, as well as induces a prothrombotic state triggering cerebral thrombosis. A notable feature is the involvement of soluble factors that promote vasculitis or thrombosis distal from the site of rash. We recently showed that plasma exosomes isolated from acute zoster patients contained significantly elevated prothrombotic and proinflammatory proteins including thrombospondin-1, caveolae-associated protein 2, coagulation factors V and XIIIA1, calmodulin 1, and transthyretin compared to matched controls. These exosomes were non-infectious but induced IL-6 and IL-8 secretion when applied to naïve primary human brain vascular adventitial fibroblasts, activated donor platelets, and induced platelet-leukocyte aggregations supporting a proinflammatory and prothrombotic state. Importantly, these exosomes persist for at least 3 months post-zoster, well after rash clearance and detectable viremia. Overall, we hypothesize that circulating exosomes during zoster and in subsequent months will promote vasculitis and thrombosis, thus providing a mechanistic basis for the increased vascular dysfunction risk preceding vascular dementia. To test this hypothesis, we will isolate plasma exosomes from individuals during acute zoster and at 7 days, 1 month, 3 months, 6 months, and 12 months post-zoster, as well as isolate exosomes from matched control individuals during a single clinic visit. Aim 1 will analyze exosome content for known vascular damaging proteins/miRNAs from matched control and zoster individuals over a 12-month period. Aim 2 will determine the mechanism(s) in which non-infectious zoster exosomes promote vascular dysfunction. Understanding how long-lasting pathogenic exosomes promote cerebrovascular disease during and after zoster, thereby accelerating vascular dementia risk, is significant because it will potentially change clinical practice with regards to duration of antiviral therapy and use of anti-platelet agents for zoster, as well as increase support for vaccination to prevent rash and stroke, as well as to decrease vascular dementia risk.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY. People with Down syndrome (DS), the genetic condition caused by trisomy 21 (T21), display a unique clinical profile across the lifespan. Early on, children with DS present stunted growth and neurodevelopmental delays. Later in life, adults with DS experience an atypical form of accelerated ageing. Furthermore, they also experience a differential clinical risk profile, marked by high risk of congenital heart defects, leukemias, autism spectrum disorders, seizure disorders, autoimmune conditions, and Alzheimer’s disease, among others. Therefore, research efforts investigating the mechanisms by which T21 causes the hallmarks of DS, as well as the factors that modify these effects, will accelerate discoveries to improve health outcomes in this population. Within this context, in 2016 we launched the Human Trisome Project (HTP), a natural history of study of DS, including deep annotation of clinical metadata, cognitive phenotyping, a multidimensional biobank, and generation of -omics datasets. The HTP has enrolled >1150 participants, >750 with T21, from all around the USA. The HTP was one of the first studies that released data in INCLUDE Data Hub, including unpublished multi-omics datasets and its biospecimen catalog. Despite these important efforts, the potential of the HTP remains unfulfilled, as it does not capture the massive diversity of the global community with T21. Therefore, we propose to develop an international component for the INCLUDE Project through the following Specific Aims: 1. To develop a network of research sites across Latin America. Led by a polyglottal and multicultural team at the Linda Crnic Institute for Down Syndrome, we recently launched the HTP – Latin America Network to support research activities in México, Colombia, Brazil, Argentina, and Chile. Each Latin America site provides massive expertise through decades of managing the largest DS clinics in their respective countries, while also contributing populations of unparalleled racial, ethnic, and cultural diversity. 2. To define the clinical and neurodevelopmental profile of Latinos with Down syndrome. Although Latinos are the fastest growing segment of the population with DS in the USA, little is known about potential differences in their developmental and clinical profiles. Therefore, we propose to complete a comprehensive annotation of demographic information, social determinants of health, co-occurring conditions, and lifestyle factors. 3. To identify biosignatures associated with differential features of Latinos with Down syndrome. We will complete a multi-omics investigation of the variable pathophysiology of DS in the Latino population, including analyses of the genome, transcriptome, proteome, metabolome, immune system, and microbiome to define associations between differential developmental and clinical features and underlying biological processes. Altogether, our efforts will provide unmatched diversity to the INCLUDE Project in terms of the population studied, investigational approaches employed, and diversity of the researchers involved.

NIH Research Projects · FY 2025 · 2024-09

Up to 33% of people report experiencing intimate partner sexual violence (IPSV). Yet, little research has examined proximal and temporal predictors of IPSV among couples. The impetus for this project is to address four critical gaps in the current scientific evidence base on IPSV: (1) the role of alcohol in IPSV, both as a predictor and an outcome, has been largely overlooked in research despite alcohol being a contributing cause and consequence of non-sexual partner violence, (2) most research on IPSV is cross-sectional, which prevents identification of causal predictors, (3) current research on IPSV fails to take into account dyadic associations (e.g., effects of each partner's alcohol use, how each partner communicates consent and refusal), and (4) there are no empirically supported theoretical models of IPSV. Using rigorous experimental and intensive longitudinal designs—the goal standard for establishing proximal and temporal relationships—the overall objective of this proposal is to determine the effect of alcohol and communication (i.e., consent and refusal) on IPSV, which will inform an etiological model for IPSV. The aims of proposal are to (1) examine the effects of one's own and one's partner's acute alcohol use and partner communication refusal (indirect verbal vs. direct verbal) on laboratory IPSV perpetration (Aim 1) and (2) examine the effects of event-level alcohol use and communication (indirect vs direct, verbal vs non-verbal) on IPSV perpetration over time (Aim 2). Across two studies, 440 couples with a history of partner violence and heavy drinking will be recruited. In Study 1, using a couple- level 4x2 design with 240 couples (n = 480 participants), couples will be separately block- randomized into eight experimental conditions corresponding to: (1) one of four couple-level beverage conditions (alcohol/alcohol, alcohol/no-alcohol, no-alcohol/alcohol, no-alcohol/no-alcohol for Actor/Partner) and (2) one of two couple-level partner sexual communication conditions (direct/indirect refusal), and then complete validated laboratory paradigm for IPSV perpetration ostensibly against their partner. In Study 2, 200 couples (n = 400 participants) will complete 26 weekly assessments measuring alcohol use, communication (indirect/direct, verbal/nonverbal) and IPSV for each day of the prior week. This proposal will inform etiological models of the association between individual and contextual factors for intimate partner violence perpetration over time. In doing so, this project will offer valuable insights that can inform future interventions aimed at reducing IPSV.

NIH Research Projects · FY 2025 · 2024-09

O’Brien R50 Research Specialist (Clinician Scientist) Project Summary/Abstract While many children with acute lymphoblastic leukemia (ALL) are cured with frontline therapy, for those who experience relapse, outcomes remain suboptimal. I am a pediatric oncologist specializing in the care of children, adolescents, and young adults with high-risk and relapsed leukemias with a research focus on clinical trials of novel therapies in both the relapsed and frontline settings. My clinical research efforts are conducted primarily through collaborative studies that are made possible by the National Clinical Trials Network (NCTN), specifically the Children’s Oncology Group (COG), where I am currently spearheading the pediatric development of inotuzumab ozogamicin as the Study Chair or Co-Chair of two large COG ALL trials, am a member of the COG Relapsed ALL Steering Committee, and am the Vice-Chair of the APAL2020SC screening trial for molecular characterization of relapsed leukemias with allocation to targeted clinical trials. Beginning in October 2023, I will serve as the COG ALL Committee Vice-Chair for Precision Medicine/Relapse. My research interests include improving our understanding of the toxicities of novel agents, identifying patients at highest risk for side effects and devising optimal management approaches for diverse patient populations based on clinical features, leukemia biology, and pharmacogenomic factors. In this era of multiple novel agents with activity in ALL including bidirectional antibodies, antibody-drug conjugates, chimeric antigen receptor T-cell therapy, and targeted agents including kinase inhibitors and modulators of apoptosis, the development of well-designed and efficient clinical trials to determine how to incorporate these agents for different populations will be critical to moving the field of ALL therapy forward over the next decade. Locally, I serve as the Medical Director of the Leukemia/Lymphoma Program at Cincinnati Children’s Hospital Medical Center with a focus on pilot studies of novel agents that, if promising, can be further developed in COG. The long-term objective of this application is to support the time needed over the next five years to complete the active COG inotuzumab ozogamicin trials (NCT03959085, NCT02981626) and to lead development of the COG portfolio of trials for both newly diagnosed and relapsed ALL.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY The main treatment of cancer is surgery, which has shown the best overall survival rate compared to radiotherapy and chemotherapy; but after the surgery, detection of lymph nodes metastases strongly influences the patient prognosis. Moreover, the most important factor for predicting long-term cancer survival is completeness of the surgical resection; but this may be difficult to achieve with microscopic disease. Unfortunately, current imaging modalities to localize cancer, such as PET, CT, and MRI, which are used pre- and post-operatively, are limited by resolution - larger than 0.5–1cm to be detectable. Consequently, current treatment failure in cancer is partly driven by lack of molecular imaging modalities to assess occult metastasis, response to therapy, or improved surgical resection techniques. Thus, detection of micrometastatic disease, achieving good resection margins at the time of surgery, and proper identification of nodal spread remain hurdles to improving surgical outcomes, treatment morbidity, and overall survival outcomes for our patients. The proposed research aims to address the limitations of current imaging modalities in cancer detection and surgical resection. By developing near-infrared chemiluminescent and fluorescent (NIR CFL) chitosan-based nanoparticles (CNPs), we seek to improve the detection of micrometastatic disease and guide image-guided surgery. These CNPs can be activated by body heat and stored at low temperatures, providing convenience and safety during use. Through the conjugation of synthetic targeting ligands, we aim to enhance tumor targeting and assess surgical margins and metastatic lymph nodes using chemiluminescence and fluorescence optical imaging. Our approach combines in vivo and ex vivo imaging modes, enabling the identification of deep anatomical locations and microscopic targets with high contrast and excellent signal-to-noise ratio. Preliminary results obtained in mice demonstrate the feasibility and practicality of our approach, showcasing the ability of chemiluminescence imaging to detect target sites over 4cm below the body surface. The anticipated outcomes of this research include advancing the combined use of optical imaging in vivo, enabling improved detection of micrometastatic disease, reducing surgical morbidity by preserving normal tissue, and providing valuable guidance for management decisions in cancer patients. By addressing the challenges associated with cancer treatment failure, incomplete resection, and nodal spread identification, this project has the potential to significantly enhance surgical outcomes, reduce treatment-related complications, and improve overall survival rates for patients.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY Head and neck cancer (HNC) remains a lethal disease despite concerted efforts to improve its diagnosis and treatment. Although smoking, drinking, and HPV infection are closely linked to HNC, emerging evidence also suggests that changes in the human microbiome are associated with HNC. However, little is known about how altered microbiota affect HNC pathogenesis or its treatment. The oral cavity, which is directly relevant to the local microenvironment in HNC, harbors one of the most complex and diverse microbiomes of all human anatomical sub-sites. However, how the oral microbiota shapes the local and systemic environment in HNC pathogenesis is completely unknown. Our long-term goal is to determine the functional role(s) and mechanisms of action of the microbiome in HNC and to translate these findings into novel preventative and therapeutic strategies. In the proposed study, we will extend and deepen previous studies of human microbiota in HNC, focusing on a prevalent form of HNC with high mortality and morbidity, oral squamous cell carcinoma (OSCC). Our Specific Aims are as follows 1) To define OSCC-associated metagenomic and metabolomic signatures in treatment naïve OSCC cases and non- OSCC controls and 2) To determine whether microbiota transfer from human OSCC cases to mouse models promotes tumorigenesis and characterize the underlying molecular mechanisms. Our approach is innovative because it represents a departure from the status quo by utilizing unique experimental mouse models and state- of-art technologies to move beyond observational human studies to delineate the molecular, cellular, and immunological mechanisms induced by the human microbiome in OSCC pathogenesis. The proposed research is significant because it is expected to advance and expand understanding of how the microbiome as a whole, as well as specific microbial taxa, genes, and pathways, impacts OSCC. Ultimately, such knowledge has the potential to be developed into effective therapies for OSCC patients, a pressing need given the significant incidence and poor prognosis of this disease.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Inhibitory synapses are crucial for maintaining correct neuronal excitability, which is important for efficient circuitry and proper brain function. Shifts in neuronal excitability have been implicated in a variety of neurological disorders, including ischemia. Based on cell-type specific vulnerabilities, the oxygen and glucose deprivation (OGD) observed in various brain regions including the hippocampus leads to differential effects that may alter neuronal long-term function. The hippocampus is particularly a vulnerable brain region that experiences either delayed cell-death in the CA1 region or resistance to delayed-cell death and shifts in long-term excitability in the CA3 region. Cell-type specific synaptic alterations to neuronal populations may contribute to these ischemic-induced changes. Even though alterations at excitatory synapses are well defined, our knowledge of alterations at the inhibitory synapse remain elusive. Inhibitory GABAA receptors (GABAARs) mediate the majority of fast synaptic inhibition in the brain. The number of postsynaptic GABAARs influences inhibitory strength; therefore, GABAAR trafficking to and from synaptic sites or the neuronal surface is an important regulator of overall inhibitory synaptic strength. During OGD, GABAARs are downregulated from the neuronal surface in hippocampal neurons. Moreover, GABAAR phosphorylation status influences the synaptic declustering or removal of receptors from the neuronal surface during OGD in hippocampal neurons. However, mechanisms that regulate these differences in either synaptic clustering or surface GABAAR expression following an ischemic insult in brain regions with varying susceptibilities remain undefined. In this project, I propose that GABAAR declustering and endocytosis mechanisms are differentially regulated in distinct neuronal populations during OGD to influence GABAAR downregulation based on region-specific vulnerability. Moreover, I hypothesize that these cell-type specific mechanisms drive increased neuronal excitability during OGD due to increase synaptic declustering and decreased surface expression of GABAARs in vulnerable neuronal populations. Based on this, I plan to investigate (i) mechanisms of synaptic GABAAR declustering and surface downregulation in hippocampal pyramidal neurons following OGD (ii) probe the temporal regulation to determine the sequential flow of events promoting GABAAR loss and (iii) use an in vivo model of cerebral ischemia to compare cell-type specific mechanisms in CA1 and CA3 hippocampus that may be differentially regulated based on neuronal susceptibility to OGD. Specifically, I plan to investigate the role of phosphatases in regulating GABAAR phosphorylation state to promote GABAAR declustering and endocytosis during OGD in both vulnerable neuronal populations. The results of this project will establish mechanisms that are specific to GABAAR downregulation in vulnerable populations during OGD, providing novel targets for future therapeutic intervention.

NIH Research Projects · FY 2025 · 2024-09

Alcohol misuse is one of the leading preventable health risks globally, contributing to over 60 different diseases and injuries. The associated morbidity and mortality impose a significant health and economic burden on society. Alcohol-induced end organ diseases, including alcohol-associated liver disease, cancer, pancreatitis, and others, contribute significantly to alcohol-related morbidity and mortality. These diseases often synergize with other epidemic diseases such as obesity, diabetes, and drug abuse. Despite advances in the field, the underlying disease etiology of alcohol-induced end organ diseases remains poorly understood. There is an urgent need to gain new knowledge of the molecular underpinnings to accelerate diagnosis, prevention, and treatment. This R13 proposal seeks support for the Gordon Research Conferences (GRC) on Alcohol-Induced End Organ Diseases in 2025 and 2027. The overarching goal of the GRC Alcohol-induced End Organ Diseases meetings is foster open and inclusive discussion of research developments, establish new scientific collaborations, encourage participation of broad representation of individuals, and propel the next generation of scientific advances. The GRC was initiated in 2017 and has held successful meetings in 2021 (delayed to 2022) and 2023. The open format of the GRC model allows for sharing significant unpublished work and provides ample opportunities for in-depth discussions with peers, new colleagues, and the future generation of alcohol researchers. The GRC on Alcohol-Induced End Organ Diseases plays a crucial role in advancing the field by providing a platform for the dissemination of key research findings, fostering collaboration, and promoting scientific progress. The unique format of the conference ensures ample time for in-depth discussions, enabling the formulation of new approaches and the initiation of collaborations. By providing a safe and respectful forum for open discussion, the conference encourages the exchange of cutting-edge research on alcohol-induced tissue injury. The introduction of breakthrough techniques facilitates a deeper understanding of the mechanisms underlying alcohol-induced tissue injuries, paving the way for innovative approaches to diagnosis, prevention, and treatment. Moreover, the conference promotes interaction between junior and senior investigators, fostering mentorship opportunities and facilitating knowledge transfer across generations. By engaging researchers across career stages and scientific disciplines, the conference fosters a broad range of perspectives and promotes active participation in the research community. This R13 grant will support travel awards to meritorious trainees and junior faculty attending the meeting. Travel award recipients will be selected based on scientific merit, with emphasis on the impact and quality of their submitted work. We envision this meeting to be welcoming to researchers of all perspectives and to significantly advance our understanding of the underlying mechanisms of end organ injuries and drive the development of effective therapeutic strategies for the diagnosis, prevention, and treatment of alcohol use disorders, ultimately improving health outcomes worldwide.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT This project will test an innovative program of automated screening using natural language processing (NLP) to improve early recognition of child physical abuse in urgent and emergency care settings. Physical abuse affects approximately 125,000 US children each year, with infants at highest risk. More than 30% of children who suffer serious abusive injuries have had prior minor injuries that might have raised the concern for abuse. Improving the recognition of these including bruises, fractures, and others, is the best opportunity to prevent recurrent abuse, escalating injury, and death. Missed abuse is especially common in emergency departments and urgent care settings, and remains a major public health problem despite efforts at education and awareness-building, routine screening by clinicians, mnemonics, clinical decision rules, and guidelines by professional societies. These interventions are resource-intensive and depend on human vigilance to recognize a condition that is rare in each setting, but which has enormous collective impact. Further, these interventions are vulnerable to bias and may exacerbate racial and ethnic disparities. Automated computer screening occurs in the background of clinical care and can overcome limitations of methods that depend on human effort. We developed and internally validated one automated screener that uses NLP to analyze unstructured narrative data within the electronic health record. External validation is needed to determine accuracy in other clinical settings and test the association of high-risk injuries with subsequent abuse. Further, testing is needed to determine the effect of the screener on racial disparities and avoidable Child Protective Services reports. If externally validated and shown to be equitable, this NLP screener could be rapidly adopted in diverse settings. We propose to externally validate this tool and accomplish these other objectives by determining statewide CPS outcomes (referrals, substantiations, services provided) for >100,000 urgent care and ED visits for infants in two large healthcare systems. Aim 1: Externally validate an automated NLP screener to identify infants with high-risk injuries in urgent and emergency care settings. We will use manual chart review as the criterion standard for visits identified by the NLP screener and other high-risk encounters. Aim 2: Determine rates of subsequent abuse for infants with high-risk injuries identified by automated NLP screening but not by usual care. We will link statewide clinical and CPS outcomes to identify abuse within 12 months for >1100 encounters with high-risk injuries. Aim 3. Determine the potential impact of NLP on racial disparities. The expected outcome of the project is the external validation of an automated abuse recognition tool to improve recognition and tertiary prevention of abuse. Our innovative methods could also be used to test other healthcare interventions to improve the recognition of abuse and other conditions.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Given the persistent disparities in perinatal health outcomes in the U.S. and the increasing utilization of fertility treatment to achieve pregnancy, the overall goal of this study is to investigate the experiences and outcomes of women from various demographic backgrounds who receive fertility treatment and achieve a live birth. Our specific aims are: 1) For fertile, subfertile, and assisted reproductive technology (ART) groups, we will quantify disparities among women in (a) maternal morbidity (based upon Centers for Disease Control and Prevention definition) and mortality during birth hospitalization through the first year postpartum and (b) infant health outcomes at birth and through the first year of life (e.g. mortality, re-hospitalizations, and emergency room use) through analysis of the population based Massachusetts Outcome Study of Assisted Reproductive Technology (MOSART) data system from 2004-2020, which includes data from the Society for Assisted Reproductive Technology Clinic Outcome Reporting System (SART CORS) and the MA Pregnancy to Early Life Longitudinal (PELL) data system (an ongoing longitudinally-linked population-based system that includes birth certificates, death records, and hospital utilization data for MA women and their infants). 2) Among women who receive ART or other fertility treatments and their infants, we will investigate the impact of individual-, healthcare-, and community-level factors associated with adverse perinatal health outcomes, through linkages of the MA Pregnancy Risk Assessment Monitoring System (PRAMS) with the MA PELL data system. PRAMS collects state-specific, population-based data on maternal attitudes and experiences before, during, and shortly after pregnancy with specific questions pertaining to receipt of fertility treatment. 3) Through qualitative interviews with women who have received fertility treatment and a live birth, we will identify facilitators and barriers to successful navigation of fertility treatment and subsequent maternal and infant care that may lead to excess risk for adverse maternal and infant health outcomes. This proposal will provide a comprehensive investigation using quantitative and qualitative methods that will inform the development of future policies and programs aimed at providing high quality perinatal healthcare to those accessing and receiving ART and other fertility treatments.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY In the United States (U.S.) an estimated 300,000 youth, ages 13-17 years, identify as transgender and, over the past decade, increasing numbers of youth have sought gender affirming care. Leading medical organizations— including the American Academy of Pediatrics, the North American Society for Pediatric and Adolescent Gynecology, the Association of American Medical Colleges, and the American College of Obstetricians and Gynecologists—consider gender affirming care in adolescence medically necessary standard of care. Youth who receive gender affirming medical care, for instance, report improved mental health outcomes sustained into adulthood. Despite these findings and medical consensus, increasing numbers of state-level policies restrict or ban medical and surgical care for transgender youth. The outcomes of these policies on transgender and gender diverse youths' psychosocial, mental health, and physical health are unknown. The goal of the proposed study is to determine the feasibility of a longitudinal, mixed methods study that assesses the outcomes of these state- level gender affirming care policies on transgender and gender diverse individuals. We will conduct a mixed- methods feasibility study and pilot with three specific aims: (1) To establish the acceptability of study instruments in the study population and the sustainability of data collection and management procedures over six-months; (2) To determine the feasibility of remote recruitment of transgender youth seeking gender affirming healthcare in U.S. with differing policy contexts, including states with criminalized bans, and those with or without restrictive policies; and (3) To assess initial, descriptive differences in our primary outcomes of interest, including receipt of desired gender affirming care, and to describe qualitative themes. In researching the impact of policy changes on the health of sexual and gender minorities, this project is significant, innovative, and responsive to NICHD priorities (NOT-HD-22-038). This is the first step in developing a timely study to address two critical knowledge gaps: (1) The quantitative and qualitative experiences of transgender individuals under the age of 18, which are underreported in the literature; and (2) The influence of state-level policies on transgender youths' psychosocial and health outcomes. The results of this feasibility study will inform recruitment strategies, data collection and instruments, and sample size estimates for a future R01.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Chronic liver diseases are a collection of disorders that result in significant morbidity and mortality worldwide. The overwhelming burden of these diseases results in the liver being the second most transplanted organ. As a result of a dramatic rise in the incidence of obesity and alcohol consumption, the rate of liver transplantation continues to climb world-wide. Despite the significant impact of chronic liver disease on society, there are no effective therapies to ameliorate disease outside of lifestyle modification and liver transplantation. Thus, a more rigorous understanding of the mechanisms that drive the progression of chronic liver disease is required to identify novel targets for therapeutic intervention. Our previous publication demonstrated a significant accumulation of T lymphocytes in the liver of individuals with Non-Alcoholic Steatohepatitis (NASH)-induced cirrhosis. However, the precise role of T cells in the progression of NASH has yet to be firmly established. The preliminary studies presented in this application have identified a yet to be appreciated role for T cell activation and clonal expansion during NASH. In this application we demonstrate that T cell clonal expansion is a common event in both humans with NASH-induced cirrhosis and mice with NASH. These are the first studies to identify T cell clonal expansions in the liver of humans and/or mice with NASH and link T cell activation and function with NASH pathology. However, the exact antigenic cause of T cell activation or if these T cells are inducing or suppressing disease progression is unknown. The studies in this application aim to answer these two fundamental questions by defining (1) the timing of T cell clonal expansion in the liver during the progression of NASH, (2) the antigens recognized by clonally expanded T cells, (3) the antigen presenting cell (APC) that drives T cell clonal expansion, and (4) if eliminating T cell clonal expansion through depletion of clonally expanded T cells or the APCs presenting antigens to these T cells results in prevention and/or resolution of disease. Completion of these studies will provide a paradigm shift in our understanding of the pathogenesis of NASH and will generate the tools necessary to define the role of T cells in the progression of other chronic liver diseases.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY On July 6th, 2023, the FDA approved Lecanemab-IRMB, the first drug with evidence of slowing the rate of cognitive decline through a reduction in amyloid plaques, the protein that accumulates in the brains of people with Alzheimer’s disease. Like all major medical treatments, there are trade-offs and alternatives that should be considered. First, the improvements in clinical symptoms are minimal. For example, Lecanemab-IRMB only demonstrated a 0.45 improvement on an 18 point scale at 18 months despite drastic improvements on imaging evidence of Amyloid. Clinically, this therapy is going to be challenging to discuss with patients. If this is like other high-cost therapies with a common indication, patients and families will likely see direct-to-consumer information and will approach clinicians hoping to receive this therapy. Situations such as this are ripe for standardized information such as a patient decision aid (DA) to help clinicians communicate effectively and accurately about the benefits and burdens and help patients and families make a well-informed decision. Shared decision making supported by patient DAs is an evidence-based strategy to improve decision making. A DA regarding dementia care could facilitate higher quality discussions to assure that patients wanting this therapy are fully informed regarding the potential benefits and harms. Using an iterative, user-centered approach with multilevel partner input, we will conduct a needs assessment among patients and clinicians while simultaneously developing and piloting a DA to support SDM for pharmacologic drugs for dementia. The project aims include: Aim 1 (year 1): Among a sample of eligible patients, caregivers, and clinicians, we will:  Aim 1a: Conduct a qualitative decision needs assessment of pharmacologic therapies for dementia.  Aim 1b: Simultaneously develop a DA based on international standards and user-centered design. Aim 2 (year 2): Using a pilot trial of 20 patients and their clinicians considering pharmacologic therapy for dementia, we will:  Aim 2a: Determine the feasibility and acceptability of a dementia DA among a diverse population of older adults and their clinicians using a mixed-methods approach.  Aim 2b: Determine the preliminary efficacy of the dementia DA on knowledge, decision conflict, satisfaction with decision, shared decision making, and care concordant with patient preferences. We aim to make sure that patients’ and families’ expectations are aligned with the expected benefits and that they are fully aware of the potential costs and harms of the new therapies for Dementia

NIH Research Projects · FY 2024 · 2024-09

Project Summary/Abstract T-helper (Th)-17 lymphocytes are central mediators of adaptive type 17 immunity, which is characterized by the expression of IL-17 isoforms including IL-17A. Decreased type-17 signaling increases severity of infections in humans and mice, but aberrant type 17 inflammation is implicated in autoimmune diseases. Effective host defense thus requires type-17 immune calibration, but mechanisms are poorly understood. A gut-lung axis has been proposed to coordinate homeostatic protection and acute host defense against infection. However, a better mechanistic understanding of this concept is needed to better understand links between the gut-lung axis and host defense. Factors that alter the gut microbiome are heterogeneous, but common settings include diet, digestive disease states such as acute infections or colitis, and transient changes due to medical treatments such as antibiotics. These changes have been studied most extensively in the context of chronic diseases such as rheumatologic disease, asthma, cystic fibrosis, COPD, and HIV. However, in homeostatic and acute infection settings, effects of the microbiome on immune defense are poorly understood. There is evidence in the literature suggesting that changes in the gut microbiome following antibiotic administration alter immune system responses to subsequent infections, including pneumonia. Prior studies have also shown that antibiotics reduce expression of IL-17-mediated host defense signals across distant organs. Since gut microbial homeostasis influences Th17 cell numbers both locally in the intestine and remotely in other tissues, I will explore how increases in sensitivity to infection are driven by the gut-lung axis. I postulate that a gut-lung axis promotes host defense through mucus barriers, the gut microbiome, and regulation of Th17 immunity. Here I propose to examine the function of the gut-lung axis mechanistically by focusing on how the gut microbiome affects Th17 lymphocyte populations in the lungs. I hypothesize that the gut microbiome promotes respiratory host defense through epithelium-derived signals that establish type 17 immune tone. I will test this in two Specific Aims that evaluate homeostatic defense (Aim 1) and acute responses to inflammation and bacterial pneumonia (Aim 2). My studies will reveal novel links between epithelial function and the establishment of Th17 immunity. There are strong correlations between the gut microbiome and immune responses in the lungs in settings of chronic diseases. However, gut-lung axis regulation of anti- bacterial defense in acute infections is poorly understood.

NIH Research Projects · FY 2025 · 2024-09

PROJECT ABSTRACT The Cole Lab’s overarching vision is to harness unique characteristics of the human genome to gain deeper insights into the impact of environmental risk factors on human disease. Our research focuses on two key areas. First, we aim to develop methods that address gaps in the derivation, analysis, and interpretation of phenotypes and their associations with markers of human health. Second, we utilize these and other approaches to identify risk factors and mechanisms related to disease, consequently driving the evolution of novel methods. These two pillars of my lab are synergistic, continually reinforce one another, and will support my lab’s sustainable growth. Over the next five years, our primary goals are to develop methods and expand resources for the broader scientific community that improve the derivation and utilization of environmental phenotypes for human disease research. One, we will improve the curation and optimization of phenotype pre-processing decisions using genetic heritability as an unbiased metric for phenotype precision. Two, we will use genetic correlation as a proxy for phenotypic correlation in non-overlapping individuals to evaluate phenotype similarity and harmonize traits across diverse groups of individuals. Third, to improve the accurate identification and interpretation of the role environmental risk factors play in human disease, we will extend traditional genetic causal inference methods (i.e. Mendelian randomization) to better handle excess cross-trait correlation and genetic instrument mis- specification commonly seen with environmental risk factors. Furthermore, we will use comprehensive sets of phenotypes in diverse, large-scale biobanks (UK Biobank and AllofUs) to develop and optimize these methods, and will make the methods and results publicly available for a broad range of scientists to utilize. Both in the short-term and long-term, we aim to expand public datasets with more comprehensive, meticulously curated phenotypes. In the next five years, we will link clinical trials with high-quality longitudinal phenotypes at the University of Colorado (CU) with data available through the Colorado Biobank, including genetics, health records, surveys, and geocoded data. This will enhance this resource for CU investigators, and specifically enable our group to investigate the effects of genetic risk on intervention response and clinical-grade phenotypes. Our efforts in the next five years will support our research focus on applied statistical genetics to identify risk factors and mechanisms related to disease, with an emphasis on metabolic disease as a model system for which our group has expertise. In the long-term, this work allows our group to identify novel research gaps for new method development. Furthermore, our applied research positions our team to best identify the challenges of uniting scientists across disciplines and skillsets and the gaps in current resources. Together this research and my long-term vision will empower our group and others to advance scientific discovery by pinpointing causal environmental risk factors for human diseases, anchoring our research in purpose.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY This proposal investigates why burns cause more severe liver damage in older individuals and devises an innovative treatment strategy to counter this damage alongside a novel diagnostic approach for earlier detection. This endeavor is crucial given the significantly higher mortality rate in older burn victims compared to younger patients, necessitating age-specific interventions. Burn injuries have been found to affect multiple organs, including the liver, which is highly susceptible to damage due to its unique structure and blood supply. This project focuses on burn-induced liver damage, a well-documented yet untreatable issue particularly prevalent in elderly populations. Our proposed studies investigate the role of MCJ (Methylation-controlled J protein), a protein that negatively regulates mitochondrial metabolism in hepatocytes (liver cells) following burn injuries in aged mice. We particularly focus on age-related factors such as increased production of pro- inflammatory cytokines, which can modulate mitochondrial respiration and potentially cause liver injury. This innovative approach will employ state-of-the-art methods for diagnosing mitochondrial respiratory dysfunction and oxidative damage, as well as assessing liver function. Our preliminary data show increased MCJ expression in the livers of aged mice postburn in settings of heightened age-related hepatic inflammation. Furthermore, in vivo MCJ silencing in hepatocytes of aged mice reduced postburn mortality. From these observations, we hypothesize that the inflammatory response after burn injury in aged subjects upregulates MCJ expression in hepatocytes, leading to reduced mitochondrial respiration and subsequent liver damage and dysfunction. We also predict that therapeutic silencing MCJ in hepatocytes can be a novel strategy to attenuate the aberrant hepatic response to burn in older subjects. To test this hypothesis in Aim 1, we will investigate how MCJ alters mitochondrial respiration in hepatocytes of aged mice postburn, potentially leading to liver damage. In Aim 2, we will examine the role of pro-inflammatory cytokines, specifically tumor necrosis factor-α (TNF-α) and interleukin-1beta (IL-1β), on postburn mitochondrial respiratory dysfunction and MCJ upregulation. Lastly, in Aim 3, we will explore a novel liver-focused treatment using small interfering RNA conjugated with N-acetylgalactosamine (GalNAC-siMCJ) for reducing postburn liver damage in aged mice by silencing MCJ specifically in hepatocytes. In parallel, we aim to evaluate the potential of MCJ (DnajC15) mRNA serum levels as a biomarker of liver damage in burn patients. This proposal holds transformative potential in human health, particularly for aging burn victims. By innovating therapeutic approaches, we aim to significantly reduce mortality rates and improve prognoses. Additionally, considering the heightened vulnerability of the elderly population to liver diseases, our research could drastically affect a broad spectrum of liver pathologies. Essentially, our work seeks to unravel the complex interplay between age, burns, and liver damage, laying a solid foundation for pivotal future research and clinical applications.

NIH Research Projects · FY 2025 · 2024-09

Project Summary A role for B-cells in multiple sclerosis (MS) pathogenesis has been implicated by clinical, histopathologic and immunologic studies. The precise role they play in pathology is debated and impact could be mediated through antibody (Ab) production, secretion of pro-inflammatory cytokines, or antigen presentation to T cells. To study Abs produced by MS CSF B cell clones, we developed protocols to produce recombinant antibodies that duplicate the fidelity of CD138+ plasmablast clones expanded in the CSF compartment of early active MS patients. We identified a subset of antibodies that bound to myelin and induced complement-mediated oligodendrocyte cell death and demyelination in vivo. These lesions spontanelusly repair and have guided independent studies to investigate the role of microlgia in remyelination. Myelin-specific MS rAbs bind to membrane lipid complexes anchored by the myelin protelopid 1 protein (PLP1) that is enhanced by sulfatide and cholesterol. We have further identified a new pathogenic MS rAb that binds myelin complexes independent of PLP1, but still causes extensive oligodendrocyte loss and CNS demyelination. A semi-quantitiative CSF binding assay employing PLP1 protein, myelin glcyolipids and choleserol have identified antibodies to PLP1 complexes in approximately 60% of MS patients but not in inflammatory and neurologic disease control CSF. Hence, we hypothesize that pathogenic myelin-specific MS rAbs encompass an array of epitope specificities targeting PLP1 protein, myelin glycolipids and PLP1 binding partners. We further suppose that antibodies to this complex contribute to type II MS lesions and represent a measurable and specific disease relevant response that can be quantified in patient CSF and serum. To test our hypothesis, we propose three complementary specific aims. In Aim 1, we will employ our quantitative PLP1 Ab binding assay to screen biobanked serum and CSF samples collected from clinically isolated syndrome (CIS) patients, relapsing MS patients, progressive MS patients, and healthy and inflammatory controls to determine the prevalence of PLP1 Abs across the MS disease spectrum and to correlate positive binding and titer to clinical correlates of disease activity. Aim 2 will evaluate the distribution of PLP1 myelin complex pathology in human MS lesions and determine association between identification of PLP1 complex antibodies from plaques and types of MS lesion histopathology. Lastly in Aim 3 we will identify and confirm the epitopic targets of MS rAbs through a combinantion of immunochemistry and studies in gene-targeted transgenic knockout mice. We will utilize our established models to independently evaluate the contributions of PLP1 and glycolipids to CNS demyelination and to develop improved immunoassays reflecting the complexity of antigen recognition. Together, our results will establish the role of complex PLP1 antibodies in disease pathogensis and should lead to improved bioassays for disease identification and reveal optimum therapeutic approaches to treat disease.