Oregon Health & Science University

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT The Primary Care Rural and Frontier Clinical Trials Innovation Center (PRaCTICe) Network Research Hub (NRH) is a partnership between the Oregon Rural Practice-based Research Network (ORPRN), the WWAMI (Washington, Wyoming, Alaska, Montana, and Idaho) region Practice and Research Network (WPRN) and their institutions’ Clinical and Translational Science Award (CTSA) programs. The PRaCTICe team has over two decades of experience partnering with 308 clinics and communities in rural settings on research, education, and technical assistance projects – including nearly 100 clinical research studies since 2019 (65 completed, 33 active). While 29% (932/3160) of the census tracks across the six Western states are in rural designated locations (RUCA 4-10), 76% of the priority year 1 clinical sites (N=29 clinics) we partner with in PRaCTICe are rural. PRaCTICe will be co-led by the Directors of ORPRN and WPRN, strengthen established partnerships with clinical sites in rural settings, and consists of four cores: Community Engagement, Recruitment & Trials, Data Coordination, and Communications & Dissemination. Together these cores will work to successfully accrue participants for existing primary care-based NIH studies and codesign future studies relevant to primary care clinics and communities in rural settings. Notably, our approach builds on established processes to support study codesign and infrastructure that extends beyond individual projects. PRaCTICe is anticipated to have a substantial positive impact in adopting and implementing research advances into clinical care, growing primary care engagement, and advancing health for all communities.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY The use of antiretroviral therapy (ART) in pregnant women living with HIV is an effective way to prevent mother-to-child transmission. However, HIV-exposed, but uninfected (HEU) children experience higher mortality from common infections and severe respiratory disease. However, the independent and combinatorial effects of HIV and ART on fetal immune cell development remain unknown. We propose to utilize the well-established rhesus macaque model of ART-treated simian immunodeficiency virus (SIV) infection to test our hypothesis that maternal SIV and ART exert differential effects on the development and maturation of the fetal immune system, resulting in dysregulated functional responses. To address the differential effects of suppressed SIV infection and ART, we will study three experimental groups of females. In the SIV/ART group, females will be infected with SIV and then suppressed with ART before undergoing timed-mated breeding. We will have two control groups: an ART-only group and an untreated/uninfected healthy pregnancy group. We acknowledge that the ART-only group is not a clinically relevant scenario, but necessary in the context of the proposed studies to distinguish the contribution of ART per se to the fetal immune phenotype exhibited by the offspring of SIV/ART dams. We do not have an untreated SIV-infected group, as this is not relevant to the vast majority of pregnant WLWH. GD130-135 fetuses will be obtained via Caesarean section and studied as outlined in the following Specific Aims. Aim 1. Determine the effects of maternal SIV/ART or ART on fetal immune system function. HEU children exhibit altered lymphocyte function and blunted responses of monocytes to pathogens. However, little is known about the effects of HIV/ART and ART exposure on lymphoid and myeloid cell activation during fetal development. Therefore, we will study the effects of SIV/ART or ART on both the innate and adaptive arms of the fetal immune system in circulation and in fetal tissues using a combination of flow cytometry, functional assays, and single-cell transcriptomics. Aim 2. Determine the effects of maternal SIV/ART or ART on fetal hematopoiesis. SIV/ART or ART may impact fetal immune system development by altering fetal hematopoietic stem and progenitor cells (HSPCs). Therefore, in this aim, we will isolate HSPCs from the fetal liver and fetal bones and determine the transcriptional landscape and functional reconstitution abilities of HSPCs from SIV/ART, ART alone, and control fetuses. Given the increasingly prevalent use of ART in women of reproductive age, this study will further our understanding of the effect of ART on fetal immune development. The ultimate goal of the proposed studies is to identify fetal immune system developmental pathways that can be targeted by therapeutics and interventions to improve immune outcomes for this vulnerable population of infants.

NIH Research Projects · FY 2025 · 2024-09

Project Summary More than 250,000 children survive pediatric intensive care unit (PICU) hospitalization each year in the United States, yet we have an incomplete understanding of recovery trajectory and modifiable factors to optimize survivorship. PICU survivors have multisystem diseases and suffer multisystem morbidities many years after hospital discharge in physical, cognitive, and psychosocial health domains. Our preliminary data indicate presence of clinically significant pain among 1 in 3 children and sleep disturbances in over 50% of children months to years after PICU hospitalization, placing them at risk for poorer long-term health outcomes. However, key knowledge gaps remain in understanding risk factors and mechanisms for persistent pain and sleep disturbances in PICU survivors, limiting potential interventions that could have broad implications for long-term recovery. For example, PICU survivors are known to suffer substantial cognitive impairments, particularly within the executive function construct, crucial to long-term academic achievement, quality of life, and psychosocial well-being. Chronic pain and sleep disturbances are potentially modifiable, and linked to worse executive function in other pediatric populations. However, we have an incomplete understanding of the impact of pain and sleep on executive function outcomes in PICU survivors. The central hypothesis of this proposal is that pain and sleep disturbances are important post-PICU morbidities that are key mediators between acute illness factors, psychosocial vulnerabilities and executive function outcomes in children after critical illness. We will conduct a longitudinal outcomes study of PICU survivors aged 8-18 years with the following objectives: 1) Identify pain trajectories and biopsychosocial risk factors for chronic pain in PICU survivors; 2) Elucidate longitudinal sleep disturbances in PICU survivors and sleep-pain associations over 12 months; 3) Test the impact of sleep and pain on EF outcomes in PICU survivors over 12-months. The study will utilize objective measures of acute illness severity, novel metrics of psychosocial vulnerabilities, and a mix of objective assessment and subjective report of pain, sleep, and executive function outcomes to comprehensively evaluate temporal and mediating relationships 3, 6, and 12-months after PICU hospitalization. This proposal is significant because it addresses the common and debilitating morbidities of chronic pain and sleep disturbances that affect thousands of PICU survivors annually. Our data will identify risk factors across a biopsychosocial framework to improve identification and treatment of pain and sleep disturbances, and aligns with the National Institutes of Health Sleep Disorders Research Plan and Federal Pain Research Strategy. This research will provide a greater understanding of sleep, pain, and executive function in PICU survivors key for designing and implementing interventions aimed at optimizing recovery.

NIH Research Projects · FY 2025 · 2024-09

Over the last decade, significant progress has been made in wearable sensors that detect biomarkers in a continuous and non-invasive manner from biofluids such as sweat. Parkinson's disease (PD) is the second-most common neurodegenerative disorder in the United States, and early diagnosis and management of the disease course of PD remains an urgent task. Among all biomarker types, the sweat metabolites, peptides, and anions can reflect both the phenotypic states of cells or organs and their dynamic responses to external stimuli, such as drug treatment. However, significant technical challenges exist in developing sweat metabolic biomarkers and wearable sweat sensors in PD monitoring: 1) lack of known biomarkers and accurate whole metabolomic and peptide profiles in PD patients' sweat; 2) the selected biomarkers need to be optimized for PD system and sensor technology; 3) current wearable sweat sensors fall short in sensitivity. This project aims to fill these gaps by proposing a novel framework for predicting and optimizing the PD metabolic biomarkers using large-scale multi-omics data, to innovate non-invasive wearable sweat sensor with high sensitivity and specificity. Specifically, the biomarker panels and sweat sensor will be developed by bridging state-of-the-art AI and nanotechnologies to assist the diagnosis and monitoring of PD, and will be validated clinically. The proposed research will provide an integrative infrastructure for developing novel PD biomarkers and non-invasive personal wearable devices. We will develop computational methods to solve a set of unaddressed questions in biomarker discovery, namely identification of PD-specific functional variations, prediction of metabolites and peptides in body fluids, and optimization of biomarker selection for personal wearable sensor (Aim 1). We will gain fundamental knowledge and technical capabilities in designing, processing, and optimizing of 2D materials-based wearable sensors for versatile applications. A physics-based, data-driven protocol will be developed to reveal the fundamental process-structure-property-performance relation for wearable sensors (Aim 2). We will develop new biomarker sets and wearable sweat sensors and test the performance in assisting diagnosis and monitoring of PD, which could be extended to other biological/disease systems (Aim 3). This proposed study will fill the gaps of disease-specific biomarker prediction and 2D material optimization in developing a clinically applicable non-invasive wearable device.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Over the past decade, there has been a growing epidemic of physician burnout, driven most notably, by Electronic Health Records (EHR), and the increasing amount of time providers spend on them. A number of solutions have been developed to untether the provider from the EHR. Prior to the COVID-19 pandemic, the two most widely adopted solutions were Speech Recognition (SR) and Medical Scribes (MS). The pandemic has worsened many of these preexisting issues with EHR, and with the broad adoption of telemedicine, creating new sociotechnical concerns. Fueled in part by this, 2 additional solutions, Virtual Scribes (VS) and Digital Scribes (DS) have begun to be employed for both in-person and telemedicine visits. We have previously performed a mixed methods evaluation of MS documenting significant barriers to successful adoption with significant variance in practice and outcomes. In spite of this rapid expansion of options, there is little information available directly to compare and contrast the ways they are co-opting provider work in the EHR, or their impact on provider activity in the EHR, medical record completion/accuracy, and safety issues. At OHSU, providers who adopt MS have baseline longer times to chart completion and significant greater amount of after-hours record completion, and use of a MS or SR had no impact on these metrics, with many, higher performing providers, worsening with MS adoption. These trends persisted and even widened with the use of telemedicine. The goal of this project is to expand upon these initial findings and use a mixed methods approach to determine the impact of EHR documentation assistance solutions on EHR use and provider workflow in ambulatory care. In Aim #1, we will adapt the Rapid Assessment Protocol (RAP) for in-person and virtual evaluation to expand on our prior work with MS to evaluate the impact of MS, VS, SR, and DS on provider workflow in the context of in-person and telemedicine visits. We will then convene a multidisciplinary stakeholder group to review these findings to come up with a series of metrics, both quantitative and simulation based, to allow for assessment of the 4 modalities. In Aim #2, we will use EHR audit logs to determine the impact of adoption of each of these 4 modalities, including data from OHSU and Medstar Health to improve generalizability, control for various EHR systems and allow for anchoring back to the qualitative data in Aim #1. In Aim #3, we will use the data from the first 2 aims, to create a high-fidelity simulation to all for direct head-to-head comparison of the 4 modalities on measures not reliably extracted from EHR audit logs including note accuracy, time-on-task, EHR navigation and patient-provider communication. In Aim #4, we will define and create a success matrix for selection of DA modalities to guide decision making on the use of appropriate DA resources for a given specialty type and location. We will also create a Documentation Assistance Dashboard (DAD) to allow for real-time, longitudinal monitoring of the various modalities across specialties.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY There is evidence from human and animal studies that support that imbalance in the renin-angiotensin system (RAS), specifically reduced ACE2 protein levels and/or activity, leads to adverse maternal and fetal outcomes. Mouse dams and concepti with loss-of-function alleles of ACE2 exhibit increased embryonic loss, reduced fetal weight, placental hypoxia, and preeclampsia-like phenotypes in dams. While the RAS has been extensively studied in the kidney and circulation, much less is known about the local uterine and placental RAS early in pregnancy and if their disruption contributes to adverse maternal and fetal outcomes. Our proposed collaborative project will address this need, drawing on my expertise in embryo transfer techniques. The main objective of this innovative application is to test the hypothesis that ACE2 plays a local role at the maternal-fetal interface modulating normal placental development and fetal growth independent of the maternal circulating and kidney RAS. To test this hypothesis, we will conduct in-depth morphological phenotyping, novel RAS protein analyses, and blood pressure monitoring by radiotelemetry to determine the cell-specific contributions of ACE2 in the placenta (Aim 1) and decidua (Aim 2). These experiments will elucidate the function of the RAS that are relevant to idiopathic pregnancy loss and pregnancy complications. Given the known differences between mouse and human placentas, we will also determine RAS gene expression, protein abundance and localization in rhesus macaque placentas, which are more structurally similar to human (Aim 3). The Aims outlined in this application will provide extensive information for the field by improving understanding of the basic RAS function in utero- placental development during pregnancy. Determining the causes of pregnancy loss, and pregnancy complications would greatly improve human health, reducing the associated physiological, psychological, and economic burdens and help us shape advances in maternal-fetal medicine.

NIH Research Projects · FY 2025 · 2024-09

Abstract: Occupational injuries and workplace accidents still occur on a tremendous scale, despite researchers’ development of successful occupational safety and health interventions for workplace outcomes. There is a particularly pressing need to 1) better investigate workplace safety in low- and middle-income countries (LMICs), and 2) bolster methods for substantially reducing accidents. Improving safety culture (SC), a robust indicator of occupational safety across various industries and countries, is an effective strategy for positively impacting workplace safety outcomes and accident reduction. Positive SC perceptions motivate employees to act safely, which leads to safer behaviors, fewer accidents/injuries, more engagement, and less turnover. However, there is a lack of empirical research in LMICs that examines the effectiveness of SC surveys and interventions, in part because the established methods in high-income countries (HICs) often use in-person training that is impractical in LMICs. For Malaysia, we will develop a mobile SC intervention program designed to improve the safety and health of workers. The practice of medicine and healthcare using mobile devices (e.g., cell phones, tablets), called “mHealth,” has been used in LMICs to effectively and inexpensively impact people’s health. Using mHealth as our foundation, we will create and test an evidence-based SC intervention program in Malaysia using instructional methods and training strategies that have been developed and practiced in the U.S. by our team. This project will address the significant knowledge gap regarding the impact of SC interventions in LMICs. Through our collaborative efforts, this project will also equip our colleagues in Malaysia with effective and practical tools, new experiences, and greater knowledge related to SC, thereby enhancing their capacity to further investigate and improve SC for workers in Malaysian organizations across high-risk industries. Our long-term goal is to improve workers’ health and safety by providing them and their organizations with effective strategies for building superior safety cultures, which they can implement without outside support. Through this collaboration, an additional goal of this project is to establish strong partnerships and build the occupational safety and health research capacity in Malaysia. R21 phase aims include: Aim 1. Translate and adapt our validated safety culture assessment/scale, other important outcome scales, and a safety audit tool for Malaysian workforces. Aim 2. Develop an mHealth-based SC intervention program for Malaysia's workforces to improve their workplace cultures. Aim 3. Pilot test and revise the mHealth SC intervention program in Malaysia. R33 phase aims include: Aim 4. Implement the revised mHealth SC intervention program in three high-risk industries to cross-validate the intervention program. Aim 5. Establish an Initial SC Benchmark for the Malaysia workforce.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Heavy menstrual bleeding (HMB) is common among adolescents and young adults with a heritable bleeding disorder (BD) and negatively impacts health-related quality of life and physical and mental health status. Effective treatment options are available that decrease bleeding and improve quality of life, but were not studied in those with BD, so we do not know if there is similar effectiveness in this condition. We also do not know the impact of treatment on diagnostic testing for BD or on improvement of iron deficiency. Given these gaps in knowledge, the primary objective of the Heavy Menstrual Bleeding Progestin Treatment in Bleeding Disorders Study is to conduct a multicenter prospective pragmatic trial, enrolling adolescents and young adults with HMB cared for at collaborating multi-disciplinary menstrual clinic sites nationally who are choosing to initiate hormonal menstrual suppression with a progestin treatment option (N=300). In Aim 1, we will compare outcomes related to (a) bleeding management, (b) quality of life, and (c) repletion of iron storage levels after six months of treatment with either 52 mg levonorgestrel-releasing intrauterine system (LNG-IUD) or oral norethindrone acetate 5 mg daily (NETA) in those with a diagnosed heritable BD. The primary outcome will be treatment success with a Pictorial Blood Loss Assessment Chart (PBAC) score <50 points by six months. Secondary objectives to assess bleeding management will include change in PBAC from baseline to 6 months, and self-reported treatment success. Quality of life outcomes will include change in validated scales (aMBQ, PROMIS-25, PROMIS-29) to assess health-related quality of life which are specific to adolescents and young adults in the setting of HMB. We will compare the change in ferritin levels at six months from baseline to determine the relative amount of benefit obtained from either treatment option. In Aim 2, we will compare outcomes after LNG-IUD in those with and without a BD, assessing rates of expulsion and method continuation between the two groups in addition to the outcomes assessed for Aim 1. In Aim 3, we will improve our understanding of the impact of HMB and iron deficiency on von Willebrand disease (VWD) diagnostic studies by comparing change in von Willebrand factor levels before and after successful treatment in those with HMB without a previous diagnosis of VWD, aiding our understanding of diagnostic parameters. Despite significant safety records, concerns about the use of hormonal medications in youth grow among U.S. families and providers. We must measure the therapeutic impact of treatment – not just on bleeding, but also on quality of life and health. This study will demonstrate the relative benefits of menstrual suppression treatment options for adolescents and young adults with HMB and heritable BDs, will provide population-specific estimates for expected benefit that can be used in treatment counseling, and will highlight the impact of treatment status on diagnostic test results.

NIH Research Projects · FY 2026 · 2024-09

The transition from adolescence to adulthood is characterized by increased rates of prescription opioid use and related problems. These rates may be even higher among emerging adults who use cannabis. National surveys show cannabis use is rising in young adults and is related to increased opioid use and opioid use disorder. However, there is a substantial gap in our knowledge of what individual risk factors might contribute to the development of opioid and cannabis co-use and related problems, and we know little about co-use in the context of pain. Identifying malleable factors that convey risk for problematic opioid use in the context of cannabis co-use would inform the development of preventive interventions that could be targeted to modify substance use risk for emerging adults in medical settings. Emerging adults receive the lowest level of preventative medical care when compared to all other developmental stages. Thus, a single medical encounter in which opioid medications are prescribed to manage acute pain offers a unique window into substance use patterns, and a unique opportunity to develop and deliver future interventions. The proposed study will utilize a developmental model of the impact of opioid exposure by legitimate prescription during emerging adulthood, with consideration for existing substance use, as well as pain and psychosocial experiences of the individual. This prospective cohort study will enroll emerging adults (ages 18-25) who either have no lifetime history of cannabis (5% of sample) or have used cannabis in the past month (95% of sample) and are prescribed an opioid for acute pain management in the course of routine outpatient healthcare. These participants will be followed closely during the 2-weeks post-prescription, and then prospectively for 2 years, with multimethod assessments to capture short- and long-term trajectories of opioid and cannabis co-use in the context of pain experiences. The central hypothesis is that pain experiences as well as patterns of opioid and cannabis use in the acute pain period will influence opioid use, cannabis use, and related problems over time. We will also examine the contributory roles of psychosocial and pain-related risk and protective factors, including the use of non-pharmacologic pain management strategies and prescription characteristics. Objective data on dispensed opioid medications will be obtained from medical records and prescription drug monitoring databases, and bioassay data will verify self-report of substance use. Daily associations between pain experiences, pain catastrophizing, negative and positive affect, use of non-pharmacologic pain management strategies, cannabis use, and opioid use will also be examined. Importantly, many of these risk and protective factors can be impacted with biobehavioral treatments. Determining which malleable risk and protective factors are most important during this developmental period will provide critical information for the design of interventions aimed at reducing substance use disorders and chronic pain in emerging adults.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Obesity in reproductive-age women has sharply increased over the past few decades and is expected to continue to rise for the foreseeable future. Besides a greater likelihood of pregnancy complications, such as preeclampsia and preterm birth, obese women are also more likely to experience severe pregnancy outcomes, including maternal death. In obese women undergoing infertility treatment, reports of poor/delayed ovarian stimulation, a higher frequency of in vitro fertilization (IVF) failure, and defects in placentation suggest that obesity negatively affects multiple reproductive processes, but the mechanisms remain largely unknown. Studying these events in humans is difficult due to ethical and technical limitations, as well as the challenge of controlling for confounding environmental factors. Through a National Centers for Translational Research in Reproduction and Infertility (NCTRI) grant, a cohort of rhesus macaque females receiving either a low-fat control diet or a high-fat Western- Style Diet (WSD) was established to study the adverse effects of obesity and diet on reproductive function. Fertility trials conducted with these females demonstrated that WSD consumption delayed the time to pregnancy and was associated with multiple placental abnormalities. Each female then underwent an IVF cycle to determine the impact of maternal diet on preimplantation embryogenesis. Blastocyst formation was significantly reduced in the WSD group and preliminary RNA-seq data from these blastocysts revealed altered expression of genes involved in critical peri-implantation processes. Considering the important role of epigenetic modifications in regulating gene expression changes during embryonic and fetal development, here we propose to investigate how maternal WSD-induced metabolic dysfunction impairs developmental programming at the molecular level and whether it can be reversed after resuming a low-fat diet. First, we will assess if there are alterations in DNA methylation, gene expression, and/or chromosome fidelity in preimplantation embryos from female macaques with increased adiposity and insulin resistance induced by WSD exposure. By examining changes that persist after a short versus long-term diet reversal, we will also determine whether it is a dietary intervention or overt weight loss that can improve IVF outcomes (Aim 1). Secondly, we will utilize the fetal thymus, spleen, liver, and kidney collected from low-fat diet and WSD-fed mothers during the NCTRI fertility trial to determine how maternal diet and metabolism influences DNA methylation and gene expression across fetal tissues. Comparing these results to the placenta, as well as maternal and cord blood, from the same subjects will help identify epigenetic signatures that are trans-generationally shared versus fetus-specific (Aim 2). Further comparison of DNA methylation and gene expression between embryos and fetuses will reveal genomic regions that are not only important for development, but also susceptible to adverse metabolic conditions. Findings from this study will provide insight into developmental reprogramming of the embryo, fetus, and placenta, as well as the reproductive consequences of stable DNA methylation changes induced by a maternal WSD.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The proposed training plan prepares Dr. Croff to develop expertise in Reminiscence Therapy, measures of cognition and social engagement, and dissemination and implementation science. These training objectives are supported by a team of expert mentors (Drs. Jeffrey Kaye, David Johnson, Kevin Duff, and Basia Belza) and a consultant (Dr. Cynthia Morris) with an established history of collaboration with Dr. Croff and with the SHARP study, the focus of the research portion of this proposal. The Sharing History through Active Reminiscence and Photo-imagery (SHARP) study engages older Black adults in reminiscence-based walking and social engagement to maintain or improve brain health and wellbeing. In groups of threes and using the SHARP walking application to access routes, GPS-linked historical images of local Black life and culture prompt conversational reminiscence as they walk within gentrifying Black neighborhoods. Dr. Croff builds upon prior SHARP testing in Portland and Seattle and upon preparatory work in Oakland to build a Learning Collaborative (aim 1) composed of research institutions and community organizations in Seattle, Portland, and Oakland. The SHARP Learning Collaborative seeks to understand needed environmental adaptations, cultural preferences, and recruitment and retention strategies to better suit the SHARP approach to each community. This will be accomplished by each site hosting a Community Engagement Studio with 12 community members. These consultative group sessions also provide information about salient points of interest to guide the development of 24 new routes and reminiscence content for Portland and Seattle, and 14 new routes for Oakland. To assess feasibility and acceptability of the SHARP approach in each city (aim 2), 12 healthy and mildly cognitively impaired older Black adults will test the SHARP approach, including adaptations, routes, and reminiscence content and their mode of delivery - the SHARP walking application. Testing will occur over a continuous period of 12 weeks in Portland and Seattle, and 8 weeks in Oakland. Through surveys and focus groups, participants will provide feedback on components of the SHARP approach and on the overall walking experience, as well as on the pre/post health measures administered (related to cognition, depression, and anxiety). Finally, we will assess fidelity to the intervention across the three cities (aims 3) via video and audio recording of intervention components and using a participant fidelity survey. Training in reminiscence therapy, cognitive and social measures, and dissemination and implementation science, and coupled with on-the-ground lessons learned from the SHARP Learning Collaborative as they test SHARP in three cities will culminate in the development of a comprehensive SHARP protocol and fidelity checklist. These two K01 products will facilitate the scalability of SHARP to these cities and beyond, and will contribute significantly to the conceptualization and development of a subsequent multisite R01 application to test the efficacy of SHARP on improving cognition and wellbeing.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Pancreatic adenocarcinoma (PDAC) is a highly aggressive and deadly form of pancreatic cancer with a poor prognosis and low survival rate. Despite advances in treatment and diagnosis, PDAC is still the third leading cause of cancer-related deaths, often diagnosed at late stages, and resistant to chemotherapy and radiation therapy due to its inherent tumor heterogeneity. The exact cause of PDAC is unknown, but several environmental and genetic risk factors have been identified. Moreover, mutations in genes such as KRAS, TP53, SMAD4, and CDKN2A have been reported as genetic drivers of PDAC progression. In recent studies, epigenetic changes have been shown to play a significant role in the development and progression of PDAC, but the underlying mediators that contribute to these alterations are largely unknown. Previous studies have established a connection between mitochondrial proteins and epigenetic modifications, contributing to the progression of tumorigenesis. Our research team has used mouse orthotopic tumor models to recently report that the mitochondrial protein, GOT2 (glutamic-oxaloacetic transaminase), supports the development of PDAC by exerting an immunosuppressive effect within the tumor microenvironment. Currently, it remains unknown whether GOT2 contributes to the metastasis of PDAC, however, preliminary data seems to suggest that GOT2 may play a role in modifying the epigenetic landscape in mouse PDAC cell line models. To investigate this further, we have meticulously designed our methods to successfully carry out the aims outlined in this proposal, utilizing in vivo mouse tumor models, as well as ChIP- and ATAC-seq experiments in conjunction with bioinformatics approaches. The overall goal of our proposed aims is to gain a better understanding of the mechanisms contributing the epigenetic dysregulation in pancreatic cancer and metastasis. We hypothesize that GOT2 is pro-metastatic and plays a role in reprogramming the epigenetic landscape, which leads to transcriptional profiles that facilitate the metastasis of PDAC. By revealing this new epigenetic role for GOT2, we hope to gain a deeper understanding of the transcriptional and epigenetic programs that enhance the plasticity of tumor cells and drive the progression of PDAC. Ultimately, this will provide new opportunities for the development of more effective treatments to address the increasing medical and societal burdens of pancreatic cancer.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Telomeres, the repetitive DNA at the ends of chromosomes, shorten during every cell division except in very exceptional circumstances; in particular, rapid telomere elongation in the embryo is essential for ‘resetting’ telomere length. Long telomeres at birth protect genome stability and are associated with good health and increased longevity, while shorter telomeres are associated with age-related diseases and early mortality. We have discovered that embryos of obese females exhibit fundamental deficiencies during preimplantation development that result in offspring having shorter telomeres, providing an explanation for the increased risk of poor health and early mortality in adult children of obese women. Importantly, we have also determined that enhancement of mitochondrial bioenergetics in the oocytes of obese mothers restored the capacity for telomere lengthening during blastocyst formation. This indicates an important but previously unappreciated molecular link between mitochondria and telomeres and is the first evidence of a regulatory mechanism by which maternal physiology determines offspring aging and lifespan. The proposed work will define the molecular mechanisms by which telomeric DNA is extended during preimplantation embryogenesis, including in a clinically relevant nonhuman primate (NHP) model, establishing completely new concepts around the developmental programming of healthy aging. We will use cutting-edge micro-manipulation and molecular assessments of genome activation to determine how oocyte mitochondrial membrane potential is linked to embryo transcriptional reprogramming. Further, we will identify actionable therapeutic strategies for ensuring the integrity of these developmental processes in physiological contexts where they are defective. As maternal obesity rates continue to rise, understanding the impact of diet and obesity on embryo telomere reprogramming has important clinical ramifications because telomere length at birth is a well-understood determining factor for future disease risk. Identifying targets and therapeutic approaches to positively manipulate this biology will provide opportunities to protect essential molecular reprogramming events at conception and during early embryogenesis that ultimately improve lifetime health.

NIH Research Projects · FY 2024 · 2024-09

Project Summary / Abstract The Medicines for Malaria Venture (MMV) recently published a “roadmap” for the types of medicines that are needed to support the long-term goal of malaria elimination and eradication. The roadmap consists of a wish list of target candidate profiles (TCP) and medicines (target product profiles, i.e., TPP). With the most recent revision to the anti-malarial target candidates and product profiles the MMV highlighted the need for identifying new rapid acting medicines for active case management while other drugs are needed for chemo-protection and chemo- prevention with long-acting molecules, and/or parenteral formulations (i.e., TCP-2) (Burrows, JN et al., 2017, Malaria Journal, 16:26). According to their updated roadmap new drugs are needed to protect populations entering areas of high endemicity during the final stages of malaria elimination. And drugs with causal liver- stage activity are needed for chemoprevention to prevent infection or outbreak of resistance during malarial seasons. This TCP has been modeled on the combination drug atovaquone + proguanil. As a potent and selective inhibitor of the parasite’s cytochrome bc1 complex ELQ-300 selectively targets Plasmodium falciparum in the blood and liver stages and even kills parasites developing in the midgut of the mosquito vector. Unlike atovaquone, ELQ-300 is a selective inhibitor of the Qi site of the targeted enzyme complex. With support from the NIH and US DOD we created a prodrug, ELQ-331, that is more effective in vivo due to improved oral bioavailability. This drug has been accepted by the MMV as a Preclinical Candidate for once-weekly dosing for disease prevention. We have now identified ELQ-596 with significantly improved intrinsic anti-plasmodial activity in vitro, enhanced efficacy in vivo in a mouse model of the disease and a more extended bloodstream half-life relative to its progenitor. This application seeks support for optimizing the structural features of ELQ-596 to provide a Next Generation of ELQs for once-monthly oral prophylaxis in humans. This would simplify the dosing regimen, improve compliance, decrease the dose and associated costs, and improve outcomes. Superior molecules will advance through a down-selection test cascade for assessment of selective potency and lack of mammalian cytotoxicity, metabolic stability, solubility in simulated intestinal fluids, resistance propensity and mode of action as well as efficacy against blood and liver stage malaria in mice. Prodrugs of superior molecules will be explored to assess for enhanced oral bioavailability and antimalarial performance over parent molecules. Scientists with expertise in the following areas make up the collaborative investigational team: medicinal chemistry, malaria, molecular parasitology, biochemistry, structural biology, and pharmacology.

NSF Awards · FY 2024 · 2024-09

Features are used to describe the characteristics of entities. For example, "age", "smoking or not", and "years of smoking" are features of a patient, which can be used to describe the patient's physical condition, and furthermore, to predict if she or he is likely to get lung cancer. A combination of features could be more helpful to the prediction, e.g., "age" minus "years of smoking" can be a new feature to indicate how early the patient starts smoking. This kind of feature combination is called feature generation. In the big data era, there exist enormous numbers of features, and it is not realistic to generate features manually by human experts. This project will build new technologies to automatically generate new features based on existing features, to better describe the objects and entities, and to gain better prediction performance. Additionally, this project aims to substantially improve the traceability, affordability, and explainability during the generation process. The developed algorithms and tools are expected to be generalized and applicable to a broad range of scientific and engineering problems, not just in feature generation, but also in other domains such as data pre-processing, social analysis, intelligent transportation systems, healthcare, and the internet of things. This project identifies three research tasks: (i) A Reinforcement Learning (RL) based approach to realize traceability. Two RL agents are used to select appropriate features, and one RL agent is used to select the appropriate operation. The policy network will be decomposed into two sub-networks, i.e., representation network and value network. Different agents will share the value network to improve training convergence. (ii) A heuristic approach to realize affordability. Information theory-based utility scores will be designed to evaluate features and feature sets, and the heuristic selection strategy will be designed in the generation process. (iii) A Large Language Model (LLM) based approach to realize explainability. The tabular data will be serialized into natural language strings, and comprehensive prompts will be designed incorporating feature generation expertise and domain expertise. The LLM can generate features with explanations by fine-tuning it with prompts. Two strategies will be proposed to compress the prompt. The proposed research will provide novel perspectives and methodologies as to how to generate new features by advancing the understanding and designing new generation strategies. They go beyond conventional generation methodologies that are highly dependent on domain knowledge This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Neurodevelopmental impairment remains the most vexing issue facing the clinical management of premature infants. Intrauterine infection and inflammation are significant causes of preterm birth potentially resulting in lifelong neurodevelopmental impairment, such as cerebral palsy, sensory and cognitive deficits and behavioral difficulties. Ureaplasma parvum is a bacterial species that is a common cause of preterm labor induced by intrauterine infection. Antibiotic treatment (Azithromycin) of intrauterine infection by Ureaplasma has been shown to delay preterm labor and improve fetal lung and fetal hemodynamic outcomes in non-human primates (NHP), which are the most clinically relevant animal model in which to study human preterm labor and fetal brain injury. However, mechanisms of neuroinflammation and perinatal brain injury, and the action of antibiotics on the fetal brain in relation to intrauterine infection remain poorly understood. Microglial polarization into pro- or anti- inflammatory phenotypes is a mechanism by which systemic inflammation may cause neuroinflammation in the developing brain. In the current proposal, we aim to investigate whether intrauterine infection by Ureaplasma parvum results in perinatal brain microglial polarization, oligodendrocyte maturation arrest and white matter injury. We will utilize our access to brain samples from a unique Fetal and Postnatal Non-Human Primate (NHP) Tissue Repository collected from preterm birth studies previously conducted in our cohort of pregnant rhesus macaques at ONPRC. Our NHP model of intrauterine UP infection represents a valuable opportunity and clinically relevant model system in which to investigate the mechanisms of perinatal neuroinflammation associated with infection-induced preterm labor across fetal and early postnatal development. In Aim 1 we will assess microglial polarization in response to intrauterine infection and inflammation and maternal antibiotic treatment in the fetal and postnatal preterm brain. Aim 2 will investigate mechanisms of oligodendrocyte progenitor cell maturation and myelination associated with intrauterine Ureaplasma infection. This proposal utilizes our team's extensive experience in studying pregnancy physiology and fetal neurodevelopment and uses novel spatial transcriptomic techniques in our unique NHP model of prematurity. Successful completion of this study, utilizing available stored NHP fetal and postnatal brain samples, will provide essential data on the mechanisms of fetal neuroinflammation that will contribute to future preclinical studies of the treatment of preterm labor and perinatal brain injury.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Pancreatic Ductal Adenocarcinoma (PDAC) is on track to become the second leading cause of cancer related deaths in the United States. This dismal prognosis is in large part due to >80% of newly diagnosed cases being advanced in stage. For the rare patients diagnosed early, stage I, the 5-year survival rate reaches over 80%. Thus, for PDAC the mortality rates can be greatly impacted by the development of an earlier detection strategy. Even though PDAC is a major health issue, there are still no effective screening tests approved for the general population and only patients with family history and/or the presence of a pathogenic germline variants (PGVs) in relevant high-risk genes, referred to as high-risk individuals (HRIs), have published guidelines recommending cancer surveillance. PDAC arises predominantly from microscopic Pancreatic Intraepithelial Neoplasms (PanIN) lesions categorized based on their morphological features as low-grade (LG), which are common and have a very low probability of malignant transformation, or high-grade (HG), which are rare and are considered to be of high risk for progression to invasive carcinoma, but the diverse routes of progression from LG to HG PanIN are poorly understood. We hypothesize that the development of a comprehensive, molecular and cellular 3D atlas that depicts the key events in the progression of PanIN to PDAC will be the first step in identifying much needed early detection and interception strategies. Importantly, our pre-cancer atlas will focus on a common subset of HRI patients in surveillance programs who carry PGVs in canonical DNA repair genes, BRCA1, BRCA2 or PALB2 (referred to as gBRCA). This genetic subtype creates a genomic instability state that supports PDAC development, yet the exact mechanisms are unclear. Through deep genetics and three-dimensional spatial multi- omics mapping with computational modeling, we propose to create a 4D model of the molecular and cellular trajectories that underly the evolution of gBRCA-driven LG to HG PanINs to malignant PDAC through three integrated aims: 1) to design a gBRCA progression atlas; 2) to determine mechanisms underlying progression; and 3) to forecast progression toward translational impact, supported by three functional units: i) Biospecimen Unit: to collect, process and annotate biospecimens, ii) Characterization Unit: to perform molecular, cellular and spatial characterization, and iii) Data Science Unit: to process, analyze, computationally model and visualize data. We have assembled an exceptional team of experts (the BRCA Forecasting PDAC, BForePC team), with complimentary, multi-disciplinary expertise in PDAC clinical pathology, computational biology and model development, BRCA deficiency and DNA repair, cancer genetics, and tumor microenvironment, along with cutting edge technology experts. We are uniting three universities, Oregon Health & Science University, Johns Hopkins University, and Indiana University, all with strong ties to the PDAC research community, the HTAN, and multiple NCI consortia. Together, we will leverage the atlas for translational detection and interception use cases poised for clinical evaluation in HRIs and to inform future studies extending to all PDAC.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Transcription factors and their coregulators are pivotal in directing gene regulation, cellular differentiation, and disease progression. Acting as molecular switches, these complexes shape cellular fate during embryonic development, maintain tissue homeostasis, and steer responses to environmental cues. However, disruptions in these interactions can lead to altered gene expression, paving the way for disorders ranging from metabolic imbalances to neurodegenerative conditions. Recognizing the need to study the regulatory dynamics of transcription factors, we developed a proteomics method called RIME. This technique immuno-precipitates epigenetic complexes using antibodies and employs mass spectrometry to pinpoint members of specific protein complexes. Nonetheless, RIME and other similar methods grapple with challenges like low reproducibility and constraints on low-input samples. Over the next five years, our ambition is to pioneer technologies that revolutionize the study of these complexes. A significant shift will be transitioning from mass spectrometry to DNA barcode methods, enhancing signal specificity from individual proteins. Our objective is to devise instruments that bypass sample limitations and inconsistencies introduced by mass spectrometry. Present methods only offer a mean signal for all interactors of a protein, even though it may participate in various distinct complexes. Through our proposal, we plan to craft DNA oligo-based techniques that amplify protein signals and discern which members coexist in the same complex. Beyond tool development, we will apply our methodologies to decipher dynamic shifts in transcription factor complexes, specifically at regulatory elements like enhancers. Our proteomic instruments will seamlessly complement state-of-the-art single-cell multi-omic methods that we are currently employing, enhancing our understanding of how chromatin accessibility, DNA methylation, and transcriptional patterns are influenced by epigenetic complexes.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The oxygen-induced retinopathy (OIR) model has been a valuable preclinical model for understanding the pathophysiology of retinopathy of prematurity (ROP) and ischemic retinopathies in general. Through ex vivo histologic evaluation of the nonperfusion area (NPA) and neovascularization area (NVA), OIR provides a quantitative model for evaluating therapeutic interventions that modulate tissue response to ischemic injury and neovascularization, both of which are common to multiple retinal vascular diseases. Advances in visible-light OCT (vis-OCT) have recently demonstrated the potential to provide not only in vivo quantitative OCT angiography (OCTA), but also measurement of total retinal blood flow (TRBF) and oxygen saturation (sO2) oximetry in preclinical rat models. In this proposal, we will develop a new wide-field (WF) vis-OCT device optimized for the rat eye to evaluate the relationship between the traditional OIR biomarkers (NPA and NVA) ex vivo and WF-vis-OCT measured TRBF, arterial and venous sO2, and NPA and NVA. We will accomplish this through three specific aims. 1) Develop WF-vis-OCT. We will build a new vis-OCT system that can provide 5- µm transverse resolution over a 130-degree field of view, by integrating a wide-field imaging probe that includes a custom contact objective lens. 2) Develop deep-learning-aided quantitative OCT angiography and oximetry. We will develop an end-to-end network for segmenting 2D and 3D capillary vasculature on vis-OCTA, which will enable the quantification of NPA and NVA in the rat OIR model. The retinal vascular sO2 and TRBF obtained from the new device will be further validated with an oxygen challenge study on normal rat retinas in vivo andamicrofluidicchipinvitro.3)EvaluationofratOIRmodelusingWF-vis-OCTbiomarkers.Inacross- sectional study design, we will first compare vis-OCT biomarkers (TRBF, sO2, NPA, and NVA) with traditional histological markers in the OIR model (NPA and NVA). We will then longitudinally evaluate two therapeutic interventions in vivo using the newly established vis-OCT rat OIR model. First, based on the hypothesis that hypoxia is the driving pathologic mechanism for NV development in phase II of the disease, we will evaluate the effect of maintaining the rat in a high oxygen environment on TRBF, sO2, NPA, and NVA. Second, based on the known therapeutic benefit of anti-vascular endothelial growth factor (anti-VEGF) in phase I OIR, we will evaluate the effect of intravitreal anti-VEGF delivery on vis-OCT biomarkers. If successful, this work will lead to improving our understanding not only of ROP pathophysiology, but will provide a novel scientific methodology for in vivo assessment of therapeutic interventions for ischemic retinopathies such as diabetic retinopathy, which often use the OIR model for preclinical investigation.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Uveitis is a common cause of blindness in the United States. The specific cause of uveitis is often unknown, making it difficult to implement targeted testing and treatment. By harnessing the power of artificial intelligence (AI) and ultrahigh-resolution optical coherence tomography (OCT), we will develop novel tools to help diagnose and classify uveitis by characterizing the distribution and type of inflammatory response within the eye. OCT will be used to image multiple areas in the aqueous and vitreous media of the eye and count the density of inflammatory cells. AI analysis will further classify cell types to aid in differential diagnosis. Protein levels in the media will be estimated based on the intensity of background scattering. Together, the cell count and protein level can be used to monitor the patient’s disease severity and response to treatment. The precision of OCT is ideal for clinical trials, where precise outcome measures can accelerate drug development and save costs. To further these goals, specific aims of the project are: (1) Develop ultrahigh-resolution OCT for imaging the anterior and vitreous media of the eye. (2) Quantify and classify inflammation in the aqueous and vitreous humor of uveitis patients. We will develop OCT-based biomarkers to aid in the differential diagnosis of uveitis and assessment of disease severity.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Many non-pharmacologic therapies (NPTs) such as massage, acupuncture, and chiropractic treatments are now covered for the treatment of back pain under Oregon Medicaid. However, racial and ethnic minorities access these treatments less often than white patients. The study includes two Specific Aims. In Aim 1, we will capture primary care NPT referral practices and other elements of the back pain visit through examination of 7 years of electronic health records (EHRs) from 8 primary care clinics, comparing NPT referral and other management practice differences across non-Hispanic white, non-Hispanic Black, and Hispanic Medicaid patients with back pain; we will also conduct brief interviews with medical directors to understand the context of care in each clinic. In Aim 2, we will explore barriers and facilitators of NPT through 1:1 in-depth interviews with Black and Hispanic patients with recent visits for back pain. Aim 2 will conclude with the integration of quantitative and qualitative data. This mixed methods study obtains information from the electronic health record (EHR) and directly from patient interviews across urban and rural primary care sites to explore factors that may influence uptake of NPT services differentially across racial and ethnic groups, including individual, clinic, and community factors.

NIH Research Projects · FY 2025 · 2024-08

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. We seek to define immunologic and transcriptomic signatures of Mycobacterium avium complex (MAC) pulmonary disease and microbiologic treatment response. Nontuberculous mycobacteria (NTM) are environmental pathogens that is present in the soil and water. MAC is the most common cause of NTM pulmonary disease, causing chronic, debilitating disease in predominantly older patients with underlying lung disease. Prior work has illustrated that incidence rates of MAC pulmonary disease are increasing substantially, particularly among women. Currently, the role of the host response in the development of disease is not well described. Treatment, when necessary, consists of 3-4 antibiotics for 18+ months. There is an urgent need to fill the critical gaps in knowledge of factors associated with disease burden and treatment response. Our lack of understanding poses significant diagnostic and therapeutic challenges. First, it is often difficult to discern colonization vs. disease caused by these organisms, and second it is difficult to predict who will benefit from therapy, or who will remain disease-free after therapy completion. There is a need for non-invasive biomarkers of disease burden and treatment response, to limit exposure to computed tomography scan radiation that is currently used to monitor disease along with acid-fast sputum culture and patient-reported symptoms. Based on M. tuberculosis literature coupled with observations of NTM disease in the setting of anti-tumor necrosis factor therapy, the Th1 response could explain NTM pulmonary disease pathogenesis in some patients. This prospective biobank enrolls patients in a sub-study of our ongoing MAC2v3 pragmatic clinical trial, as well as others who meet American Thoracic Society/Infectious Disease Society of America clinical, radiographic, and microbiologic disease criteria and are starting treatment for MAC pulmonary disease. This builds on our existing NTM Clinical Trials Network infrastructure and extensive experience developing and managing the Northwest NTM Biobank. The Biobank has supported collaborations between MPIs Drs. Winthrop and Lewinsohn to conduct preliminary feasibility studies of immune correlates of disease. MPIs Drs. Lewinsohn and McWeeney have a longstanding history of collaboration. During Years 1-4, we will collect blood and sputum samples longitudinally from patients initiating treatment for MAC pulmonary disease and conduct Aims 1A and 2A. During Years 3-5 we will complete Aims 1 and 2. In this setting we will conduct high throughput spectral flow cytometry analysis of peripheral blood mononuclear cells to identify an immune signature of disease and microbiologic treatment response. Similarly, we will conduct whole blood next generation RNA sequencing to identify an RNA transcriptome signature of disease and microbiologic treatment response. Ultimately, the development of a disease signature and biomarkers predictive of treatment response could help better guide therapeutic and diagnostic decision-making with regard to MAC pulmonary disease.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Allogeneic hematopoietic cell transplantation (HCT) is an essential therapeutic procedure used to enhance the outcome of patients with acute myeloid leukemia (AML). Recent advances in alternative donor transplantation and supportive care have allowed for a greater number of older adults and those with co-morbidities to pursue potentially curative transplant procedures. In addition, the emerging era of targeted therapy allows for the development of novel disease remission induction regimens as well as transplant conditioning regimens that can enhance outcomes. However, relapse after allogeneic HCT remains a major barrier to successful outcomes, with recent data suggesting that up to 50% of allogeneic HCT failure and mortality relate to persistence or relapse of underlying disease. Recent advances in the detection of minimal/measurable residual disease (MRD) have enhanced the capacity of identifying some patients at high risk of relapse after HCT. Using information obtained by cytogenetics, fluorescence in situ hybridization (FISH) and genomics, successful outcomes can still be achieved in a subset of myeloid malignancy patients transplanted in the setting of morphologically active or MRD-detectable disease at the time of transplant. Recent observations have demonstrated that more intensive remission induction chemotherapy followed directly by allogeneic transplantation can provide successful outcomes for patients with poorly responsive, unfavorable AML after primary induction treatment. This proposal will investigate the benefit of such aggressive intensive induction followed by immediate transplantation with myeloablative conditioning for patients with intermediate and high risk AML, when compared to current standard approaches. This effort will determine whether augmented remission induction therapy with early transplantation using sensitive MRD assessment before and after transplant for myeloid malignancies can be used to improve HCT outcomes with the most efficient health resource utilization. In so doing we hope to establish the importance of more intensive induction chemotherapy in creating opportunities for the best long-term transplant outcomes for the AML patient. Sensitivity of novel MRD assessments will be available for study that could determine the need for novel interventions for post-HCT patient management using a variety of emerging cellular and/or pharmacologic-targeted approaches.

NIH Research Projects · FY 2026 · 2024-08

Project Summary/Abstract The opioid epidemic is a persistent public health crisis in the United States and abroad, historically dominated by the misuse of prescription opioids and heroin; however, those illicit substances have been overtaken by a surge in the synthetic opioid fentanyl. Fentanyl’s unmatched potency and addictiveness makes it more difficult to treat with pharmaceuticals that act on opiate receptors directly; as a result, opioid overdose deaths in the U.S. skyrocketed to more than 80,000 in 2021. Novel therapeutic approaches to manage this crisis are urgently needed, making understanding basic opiate signaling mechanisms in the brain critically important. A potential target to mitigate opioid use disorder (OUD) are the cannabinoid receptors (CBRs), which are co-expressed alongside opiate receptors in the brain’s reward centers. Recent studies indicate that CBRs can modulate opioid-reward behaviors; however, our understanding is challenged by multiple CBR subtypes that have dynamic and overlapping expression in the brain. Antibodies for CBRs can be nonspecific and provide only a snapshot of receptor localization, and conventional approaches to manipulate CBRs have poor spatial and temporal resolution: Genetic knock-outs are prone to compensation, while microinjection or wash-on of hydrophobic cannabinoid ligands suffer from poor kinetics and don’t distinguish between CBRs on overlapping circuits. To delineate the role(s) of CBRs in opioid reward and as a potential therapeutic target for OUD, there is a critical need for new tools to probe their localization and manipulate their signaling with high resolution. The long-term goal of my lab is to build chemical tools for deconvoluting the molecular components of substance use disorder, and I have pioneered several chemical biology technologies to study CBRs with enhanced spatiotemporal precision. In collaboration with Erick Carreira’s lab, we developed fluorescent probes that label CB1 and CB2 cannabinoid receptors with a fluorophore. These overcome the poor selectivity of antibodies and can be applied to live-cell imaging experiments. We also developed chemical photoswitches allowing acute reversible optical manipulation of CB1 and CB2, which permits control of downstream signaling by a precise optical stimulus. The objective of this proposal is to expand our cannabinoid chemical biology toolkit towards the atypical cannabinoid receptor GPR55, and then apply these tools in ex vivo and in vivo models of opioid administration. We will explore the interactions between cannabinoid and opiate receptors in the ventral tegmental area (VTA), a critical hub in the brain’s reward system. This proposal has a strong scientific premise built on our published studies, preliminary data, and a careful review of the literature. It is innovative because it applies our state-of-the-art tools to illuminate basic signaling mechanisms relevant to opioid reward and could generate novel therapeutic avenues for OUD. This study holds significant implications not just for understanding the basic neurobiology of addiction, but also for the development of next-generation therapies for one of the most pressing public health crises of our time—the opioid epidemic.

NIH Research Projects · FY 2025 · 2024-08

Summary Due to advances in neonatal care, most preterm infants no longer develop cerebral palsy from hypoxic-ischemic white matter injury (WMI). However, up to 50% develop gray matter associated cognitive and learning disabilities by school age that persist throughout life. These developmental disabilities are commonly associated with a systemic hypoxia (Hx) episode arising from lung immaturity, acute respiratory failure or other common complications of prematurity. We have developed a preterm-equivalent mouse model which delivers a single mild clinically relevant episode of Hx at P2. Our preliminary findings support that Hx impairs learning and memory by disrupting hippocampal: (1) neuronal structural complexity; (2) gene expression; (3) synaptic plasticity/intrinsic excitability; and (4) fear-conditioning responses in juvenile mice exposed to Hx. We hypothesize that neonatal Hx is sufficient to persistently disrupt neuronal maturation without causing significant neuronal loss or white matter injury (WMI). In aim 1, we will determine if Hx disrupts hippocampal maturation independent of acute or delayed cerebral inflammation, neuronal degeneration or WMI. We will determine how Hx disrupts CA1 neuronal dendritic arbor maturation and spine density using innovative super-resolution light microscopy integrated with analysis of synaptic transmission, long-term potentiation (LTP) and intrinsic excitability. We will determine if these changes occur independently of hippocampal neuronal or glial cell death. Since WMI is common in preterm infants, we will determine if CA1 neuronal dysmaturation occurs independently of WMI. Aim 2 will build upon preliminary data showing that Hx causes persistent gene transcriptional changes in mouse hippocampus at P16 and P30. The hippocampal response to Hx involved regulators of excitatory and inhibitory synaptic transmission, synaptic plasticity and epigenetic regulators, all integral to learning and memory. To define novel molecular mechanisms of hippocampal dysmaturation, we will take an unbiased approach using single nucleus RNAseq to determine cell type-specific early and late gene expression changes arising from Hx at P2. In aim 3, we hypothesize that disturbances in synaptic transmission at CA3-CA1 synapses disrupt hippocampus-dependent memory mechanisms in young adult mice exposed to Hx as neonates at P2. We will determine the effects of Hx on excitatory and inhibitory synaptic activity and on the subunit composition of glutamate receptors that regulate learning and memory in neonates. We have identified that Hx disrupts the action of several key modulators of excitatory synaptic activity. We will focus on the synaptic potassium channel SK2, which acts as a negative feedback regulator to limit synaptic depolarization by NMDAR and AMPAR. To further define a mechanistic role for glutamatergic synaptic activity in disrupted LTP, we will determine if an allosteric AMPA receptor agonist (ampakine) delivered in vivo will strengthen synaptic transmission/LTP in vitro. Neurobehavioral testing using hippocampal memory paradigms will facilitate our long-term objective to develop rational therapies to prevent or reverse the chronic effects of Hx on maturation of hippocampal learning and memory mechanisms.