University Of Washington

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT An estimated 13% of pregnant individuals in the US experience depression perinatally (in pregnancy and/or up to 1 year postpartum), making depression one of the largest contributors to maternal morbidity. Perinatal depression is associated with adverse effects for both the perinatal person and their child. Risk of perinatal depression is inequitably distributed, disproportionately impacting low-income, racial minority individuals and those who have had adverse life experiences. The US Preventive Services Task Force (USPSTF) recently endorsed counseling interventions as moderately effective at preventing perinatal depression and recommended referral of all at-risk individuals to cognitive behavioral therapy (CBT) or interpersonal therapy. However, there remain significant barriers to accessing facility-based interventions for at-risk individuals, including lack of financial resources, time, transportation, or social support to attend in-person services. These barriers may be especially heightened in those individuals at highest risk of perinatal depression. Some access barriers may be overcome by using mobile technology to deliver evidence-based interventions remotely. We previously developed and pilot-tested a digital group adaptation of the evidence-based CBT intervention, the Mothers and Babies course. This digital intervention, named IMAGINE, was developed based on formative input from perinatal individuals and their healthcare providers. IMAGINE is a manualized 10- to 12-week intervention delivered to groups of 6-10 individuals, using asynchronous, interactive, multimedia messaging and an optional weekly synchronous group video call. IMAGINE’s strengths include its fidelity to an evidence-based intervention and inclusion of facilitator guidance and peer support to maximize efficacy. In this pilot effectiveness trial application, we propose to engage a broad range of stakeholders including perinatal individuals at risk of perinatal depression as well as service providers, administrators and payers to complete the following specific aims. In Aim 1 we will use a human-centered design approach to refine IMAGINE and prepare for its delivery as part of routine perinatal mental health services. We will then conduct a pilot randomized trial to (Aim 2) obtain preliminary data on IMAGINE’s engagement of CBT mechanistic targets and effectiveness in preventing perinatal depression, and (Aim 3) obtain preliminary data on implementation outcomes (acceptability, appropriateness, usability, feasibility) and their drivers. This proposal addresses an urgent need to improve access to evidence-based interventions to prevent perinatal depression among those at highest risk. Completion of these aims will generate preliminary data, study tools, and a participatory group of stakeholder advisers to enable pragmatic evaluation of IMAGINE’s impact on perinatal mental health in a future R01.

NIH Research Projects · FY 2026 · 2024-02

Project Summary/Abstract Asthma is an inflammatory disease of the airways characterized by acute, intermittent and recurrent episodes of inflammation that can be induced by a specific allergen. CD4+ T cells contribute to this process by producing the Type 2 cytokines IL-4, IL-5, and IL-13 and inducing B cell production of IgE in response to T cell receptor (TCR) recognition of allergen peptides bound to MHCII molecules on host cells. In both murine models of disease and asthmatic patients, quiescent allergen peptide:MHCII (pMHCII)-specific CD4+ memory T cells can persist in lungs and lymphoid organs long after resolution of inflammation. Upon subsequent exposure to allergen, CD4+ memory T cells rapidly drive asthma-induced immunopathology making these cells attractive targets for allergen- specific immune modulation. Little is known, however, about the function and maintenance of Th2 memory cells that orchestrate the asthmatic response due to the challenge of tracking small populations of CD4+ T cells that express allergen pMHCII: specific TCRs. To address this lack of knowledge, we produced an MHCII tetramer containing a peptide from the Der p1 protein of the house dust mite (HDM), Dermatophagoides pteronyssinus, the most common cause of atopic asthma. Using this tetramer and a novel magnetic bead-based cell enrichment method of our design, we have found that IL-2 dependent, functionally heterogeneous populations of Th2 tissue resident memory cells reside in the lung for long periods of time. The central hypothesis of this application is that these distinct yet synergistic populations of Th2 “effector Trm” and B cell helping “Tfh Trm” contribute to asthma pathogenesis in unique ways. The goals of this proposal are to identify the molecular and cellular mechanisms that lead to Th2 Trm cell heterogeneity and determine how these cells persist in different regions of the lung. This innovative approach could provide the means for targeting specific pathologic functions of memory Th2 cells by immunotherapy or eliminating them altogether.

NIH Research Projects · FY 2026 · 2024-02

SUMMARY Tuberculosis (TB) is caused by the bacteria Mycobacterium tuberculosis (M.tb) and was responsible for 1.6 million deaths in 2021. The only licensed TB vaccine, the Bacillus Calmette-Guérin (BCG) vaccine, protects infants and children from disseminated disease but provides only partial protection in adult pulmonary disease. T cells are known to be critical in the host response to M.tb infection, but the mechanisms of this protection are not fully understood and further research is needed to develop new, effective vaccines. The first category of T cells are classical T cells that recognize peptide antigens presented by major histocompatibility (MHC) molecules and include two subsets, helper T cells that express the CD4 coreceptor (CD4 T cells) and cytotoxic T cells that express a CD8 coreceptor (CD8 T cells). The second category are nonclassical T cells that recognize non- peptide antigens via MHC independent mechanisms and consist of several distinct subsets. Two subsets of nonclassical T cells are CD1a/b/c restricted T cells and T cells that express a CD8 coreceptor (CD8 T cells). CD1a/b/c T cells recognize mycobacterial lipids presented by CD1a, CD1b, and CD1c molecules and have exhibited antimicrobial effector functions in response to M.tb infection. In contrast, the role of CD8 T cells in M.tb immunity is unknown. Antigen independent activation of T cells, termed bystander activation, occurs via proinflammatory cytokines and has been observed to have a protective effect in cancer and infection. Specifically in M.tb infection, bystander activation of noncognate memory T cells has shown a host protective impact by decreasing bacterial burden. CD8 T cells have exhibited higher levels of bystander activation compared to their CD8 classical counterparts. In contrast, the ability of CD1a/b/c restricted T cells to undergo bystander activation is unknown. Thus, CD1a/b/c restricted T cells are implicated in M.tb immunity, but their susceptibility to bystander activation has not been explored. Conversely, CD8 T cells are known to undergo bystander activation but their role in M.tb infection has not been established. In Aim 1, we will determine the ability of CD1a/b/c restricted T cells to undergo bystander activation, explore the underlying signaling mechanisms of this activation, and evaluate the ability of bystander activation to induce anti-bacterial effector programs during M.tb infection. In Aim 2, we will define the role of bystander activated CD8 T cells in M.tb infection, defining the antigen-independent functions of the subset on granuloma formation and bacterial control. Our proposal aims to fill the corresponding gaps in knowledge of the role of bystander activated CD1a/b/c restricted T cells and CD8 T cells during M.tb infection. In addition to potentially illuminating a novel pathway to target for M.tb vaccines, these studies will more comprehensively analyze T cell activation in M.tb immunity.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT This project develops new experimental and computational methods for DEER (double electron- electron resonance) spectroscopy. DEER is a biostructural technique for the quantification of protein conformational landscapes and protein motions on the nanometer scale. Protein motions are crucial for many key molecular processes at the basis of human life and disease. Therefore, DEER provides important insights that contribute to the knowledge base necessary for drug development. In combination with X-ray crystallography, cryo-EM, NMR, FRET and others, DEER is part of a complementary set of integrative experimental biostructural tools. It is especially important for the study of membrane proteins. Several major barriers exist in the field: the lack of integrated analysis and modeling tools for biomedical researchers, and the lack of experimental approaches for studying proteins in their native cellular environment. This project directly addresses these issues as it aims to (a) develop methods and tools based on Bayesian statistics and deep learning for the rigorous and reproducible analysis of experimental DEER data; (b) create advanced computational approaches that utilize DEER data for modeling proteins; (c) develop methodology based on noncanonical amino acids for labeling proteins directly in their cellular environment; (d) advance a rapid freeze quench approach to measure conformational dynamics down to the sub-millisecond time scale. Overall, the goal of the project is to significantly expand the scope of DEER by providing innovative approaches to data analysis, modeling, and in-cell and time-resolved measurements. This will enable the study of the structure and dynamics of larger and more complex proteins and protein assemblies in the cellular environment. This is of increasing importance in biomedical research.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY Research demonstrates that substance-use disorder, including opiate use disorder, affects one's decision making and reward processing resulting in impaired goal-directed behaviors. However, there remains a lack of understanding of the neurobiology that underlies persistent reward-seeking despite value changes that would normally alter goal-directed behavior. The neuropeptide S (NPS) system, made up of the peptide and its receptor (NPSR1), drives reward-seeking behaviors, however the underlying mechanism is not understood. We generated both NPS-Cre and NPSR1-Cre driver mouse lines for accessing and examining the key circuit components of NPS/NPSR1-mediated behaviors. I isolated a population of NPS-containing cells adjacent to the locus coeruleus (LC) and found that these neurons project to the orbitofrontal cortex (OFC), a region with dense expression of NPSR1. I found that these neurons respond to delivery of reward-predictive cues as well as consumption, but to a lesser extent. I then preliminarily tested the effects of oral fentanyl self-administration on this population and uncovered a bidirectional response to delivery of the conditioned stimulus (enhanced activity), and fentanyl reward (quiescence). In this proposal, I first aim to determine the dynamics of OFCNPSR1 neuronal ensembles during natural- and drug-seeking behaviors (Aim 1) using two-photon microscopy. This aim seeks to further investigate OFCNPSR1 activity during self-administration of fentanyl reward as well as how devaluation of either a natural or drug reward is modulated by OFCNPSR1 activity. My second aim is to examine how hindbrain NPS neuron activity and neuropeptide release impact OFC encoding during reward-seeking behaviors (Aim 2). This aim serves to close this gap in knowledge by using multi-site fiber photometry to simultaneously image hindbrain NPS, and OFC neuron activity during reward-seeking behaviors. Finally, in the R00 portion of this grant, I will expand the scope of this study by determining the sufficiency of the OFCNPSR1-Basolateral Amygdala (BLA) projection in driving drug-seeking behaviors (Aim 3). I found that OFCNPSR1 neurons are connected with the BLA. In addition, the BLA expresses NPSR1, however, the source of endogenous NPS to the BLA is unknown as well as any modulation the OFCNPSR1 neurons may provide to this region. This aim serves to uncover the source of BLA NPS as well as investigate the role of BLANPSR1 neurons in reward-seeking behavior and how this activity may be affected by OFCNPSR1 projections. I will utilize a series of complimentary cutting-edge neuroscience techniques to dissect the role of the OFCNPSR1 neurons in drug- seeking behavior. The central goal is to aid in future investigation of whether NPS signaling represents an important avenue for altering drug-seeking behavior. This proposal provides substantial new training in SLM, in vivo calcium imaging of network and single-cell activity, neuropharmacological and molecular viral manipulations, and career development skills for transition as an independent academic investigator.

NIH Research Projects · FY 2025 · 2024-02

Substance use disorders (SUD) are characterized by pathological behavioral engagement with rewarding stimuli despite negative outcomes. There has been a concerted effort to identify the neural correlates of reward receipt, as well as the systems-level processes that govern chronic cannabis use disorders, a growing problem in the US. There remains a lack of mechanistic understanding of the intrinsic neurobiology which underlies behavioral engagement, a fundamental process in drug seeking behavior. As such, the primary goal of this pathway to independence proposal will be to receive in depth neuroscience training, facilitating new skills in a series of cutting-edge approaches which will provide a deeper understanding of the underpinnings of neuromodulatory regulation of engagement in reward-seeking behaviors, and how these mechanisms are altered with chronic cannabinoid use. The Nucleus Accumbens(NAc) represents an important reward network hub known to regulate reward engagement and the reinforcing properties of drugs of abuse. The NAc receives dense innervation from the Paraventricular Thalamus(PVT), which receives visceromotor and interoceptive inputs from the hypothalamus and hindbrain, and projects widely across the forebrain to regulate a variety of motivated behaviors. The PVT is a highly heterogenous structure, and recent studies examining the PVT-NAc circuit have shown conflicting results, partially driven by a lack of genetic and anatomical specificity within the PVT. The neuromodulatory peptide neurotensin(NTS) is selectively expressed in the anterior PVT and our preliminary data demonstrates that these neurons send excitatory projections to the NAc, which are enriched with the cannabinoid 1 receptor(CB1R) and tightly regulated via endogenous cannabinoid(eCB) signaling. In this proposal, we first aim to understand how eCB modulation of aPVT projections to the NAc regulates behavioral engagement. Our preliminary experiments demonstrate that aPVT projections to the NAc are inhibited during behavioral engagement and excited upon disengagement, in both rewarding and aversive contexts. In tandem with our electrophysiology data demonstrating retrograde eCB inhibition of PVT terminals, we will determine when and how eCBs are released in vivo to modulate aPVT input to ultimately affect behavioral engagement. Our second aim will be to determine how the activity of projection defined NAc neurons is causally linked to eCB production and reward seeking behavior. In this aim, we will use 1-photon imaging to record the activity of specific aPVT-NAc projection ensembles. We will multiplex this high-resolution neural data with novel machine learning behavioral tracking algorithms to directly correlate neural activity with discrete aspects of reward seeking behaviors. Finally, in the R00 portion of this grant, I will expand the scope of this study to other aPVT output regions to determine how the aPVT regulates cannabinoid seeking behaviors through divergent efferent projections across the forebrain. The training received under this proposal will facilitate my transition to heading my own independent academic research laboratory focused on decoding neuromodulation in SUD.

NIH Research Projects · FY 2026 · 2024-01

Project Summary/Abstract The potential for devastating consequences of cryptosporidiosis or giardiasis diarrhea in immunocompromised patients and malnourished children emphasizes the need for a single effective therapy that could be used syndromically where diagnosis may be delayed or uncertain. This will also be applicable for asymptomatic presentations of both diseases. The etiologic parasites colonize and reproduce in the small intestines of mammalian hosts where they are associated with epithelial cells microvilli. Hence, potential for finding dual therapeutic is high based on common anatomical site of infection. Protein kinase inhibitors have attracted considerable attention as potential therapeutics since a number of them have been released as drugs in recent years and many are in various phases of clinical trials. In a semi-High throughput screening of MMV Pathogen box and Celgene Global Health programs library, we identified dual hitting kinase inhibitors from 2 chemical scaffolds. The scaffolds offer new medicinal chemistry opportunities for development of effective multi-parasite therapeutics against cryptosporidiosis and giardiasis. This research proposal capitalizes on these preliminary findings to hypothesize that effective single agent therapy that could be used syndromically for treatment of either cryptosporidiosis or giardiasis is possible. Analogues of the 2 scaffolds already in hand and newly synthesized in Aims 1a will be screened in Aims 2a to determine in vitro efficacy in blocking parasites growth using direct phenotypic screening to develop structural activity relationship. We will also be collecting other standard in vitro ADMET and physicochemical parameters such as solubility, stability in simulated gastric/intestinal fluids, and mouse/human microsome stabilities, in-house hERG assay etc as Go/NoGo filters. Criteria for moving forward to in vivo studies will include dual parasites killing efficiency, selectivity, static vs. cidal, stability in both gastric fluids and gut Phase 1 metabolism, and ease of medicinal chemistry development. Since necessary factors for clinical relevance have not been established in these scaffolds, we will use early leads from Aims 2b to define the PK/PD properties necessary for optimum animal in vivo efficacy in Aim 2b. We will perform further in vitro safety profiling for early leads focused on further improving selectivity including quantitative proteomic analysis of offtarget safety kinases, cytochrome P450 isoenzyme activities and metabolite identification in Aims 1c. Using medicinal chemistry based iterative reasoning to synthesize analogs, we will do lead optimization for efficacy, selectivity, PK/ADMET properties, in vivo efficacy and resistance under Aim 1b. In Aim 3; optimized leads will be evaluated for clinical usefulness with experiments to define any liabilities related to CYP activities, mammalian kinome profile, AMES test, micronucleus assay and a safety panel of human receptors and ion channels. Other experiments will include final toxicology studies, additional resistance studies and metronidazole combination studies. The product of this research study would be useful as parasitic diarrheal therapeutic with the potential for broad spectrum activity. It will have significant public health benefits in many developing regions of the world.

NIH Research Projects · FY 2026 · 2024-01

Men who have sex with men (MSM) are at high risk for gonorrhea and chlamydia in Kenya, where nucleic acid amplification testing (NAAT) is not feasible and most infections therefore go undiagnosed. While development of low-cost point-of-care diagnostics is an urgent priority, low-cost and feasible interventions to control sexually transmitted infections (STI) among MSM are needed now, as we await technology advance. In 2011, the WHO recommended periodic presumptive treatment (PPT) of Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) infections for MSM at high risk for HIV acquisition due to condomless anal intercourse with multiple sex partners or a recent STI exposure. More recently, trials in well-resourced settings have demonstrated the efficacy of doxycycline post-exposure prophylaxis (doxyPEP) at reducing NG, CT, and syphilis infections among high-risk MSM. In this R01 application, we propose a rigorous study to evaluate the impact and cost-effectiveness of WHO-recommended PPT versus doxyPEP, compared to standard syndromic treatment, among Kenyan MSM. Our highly productive research team, including collaborators from the University of Washington, Aurum Institute, Nyanza Reproductive Health Society, and Partners for Health and Development in Africa, has expertise in clinical STI care, epidemiology, research with MSM populations, and intervention trials (MPI Graham and Sanders, co-Is Otieno and Kimani), NG and CT diagnosis and testing for antimicrobial resistance (AMR) (co-I Soge), infectious disease modeling (co-I Hamilton), and costing of interventions (co-I Sharma). This study aims to (1) evaluate the effectiveness and impact on AMR in NG of two interventions: WHO-recommended PPT given every 3 months and doxy-PEP taken 24-72 hours after condomless sex, compared to standard syndromic treatment, for reducing STI burden among Kenyan MSM; (2) assess the acceptability, feasibility, and safety of implementing WHO-recommended PPT and doxy-PEP compared to standard care among providers and patients; and (3) model the health and economic impact of scaling up WHO-recommended STI PPT and doxyPEP compared to standard of care on STI control among MSM and their partners in Kenya. We will conduct an open-label randomized clinical trial with 2900 participants to evaluate these two interventions versus the standard of care assigned in a 2:2:1 ratio, with 18 months of follow-up and rigorous culture-based and molecular analysis of AMR in NG at three MSM-friendly research clinics in Kenya. We will use multidisciplinary science to measure the acceptability, feasibility, and safety of these two interventions, using a conceptual model based on Proctor’s Implementation Science Framework. Finally, Aim 1 and 2 results will inform parameters to update a stochastic model of STI transmission and cost- effectiveness analysis to project the impact of scaled-up STI PPT and doxyPEP in Kenya. This work will provide the critical data needed to inform guidelines and improve STI control among these key populations in sub-Saharan Africa and other resource-limited settings.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY/ABSTRACT This F31 project aims to close gaps in knowledge that could inform vaccine policy and facilitate implementation of maternal influenza vaccines in low- and middle-income countries (LMICs). Influenza causes 3-5 million cases of severe illnesses and 290,000-650,000 deaths globally each year. Pregnant people are at increased risk of influenza morbidity and mortality, making vaccination a high priority in this population to protect themselves, their fetuses, as well as their infants via maternal antibodies. Although influenza mortality rates are estimated to be higher in LMICs, empiric data on the influenza disease burden are limited. In addition, the impact of maternal influenza vaccination on infant growth has not been evaluated while accounting for bias induced by undefined infant growth outcomes due to fetal or early infant death. Best practices for implementing maternal immunization programs have also not been established including the optimal timing of vaccination. Moreover, potential costs must be considered since LMIC health budgets often need to prioritize which interventions to implement due to resource constraints. The proposed F31 research project leverages data collected between 2011-2014 as part of a maternal influenza vaccine RCT in Sarlahi District, Nepal (NCT01034254) among pregnant people (18-40 years) and their infants who were followed through 6-months postpartum. The parent study included enrollment visits, weekly household-based active surveillance for respiratory pathogens, illness surveys on symptoms and care seeking behaviors, maternal/infant birth assessments, monthly maternal comorbidity assessments, and infant anthropometry visits at birth and 6 months to measure weight (g), length (cm), and head circumference (cm). Using log-binomial regression methods, Aim 1 will assess the impact of influenza vaccination during pregnancy on maternal influenza disease severity among pregnant participants (n=3693). Aim 2 will characterize the effect of maternal influenza vaccination on infant z-scores at birth and 6 months of age accounting for outcomes truncated by death among all singleton births using multinomial logistic regression (n=3520). Aim 3 will evaluate the optimal timing and cost-effectiveness of administering seasonal influenza vaccine during pregnancy using an existing decision tree model to project the potential impact of maternal influenza vaccine timing strategies (second versus third trimester compared to no vaccination) and calculate incremental cost- effectiveness ratios. Data generated from this project will expand the current maternal influenza vaccine evidence base in LMICs, which may motivate changes in vaccine policy, implementation, and investment decisions. As a long-term goal, the F31 study team seeks to contribute to reducing disparities in global influenza vaccine access so that all communities can benefit from this critical public health intervention. This research plan will provide the F31 candidate rigorous predoctoral training including (1) performing advanced analyses in infectious disease epidemiology; (2) developing content-area expertise in maternal immunization; (3) acquiring methods to evaluate vaccine cost-effectiveness; and (4) gaining further experience in research dissemination.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT This F31 proposal details a 3-year research and training plan that will launch the applicant’s career as an independent clinical investigator focused on mind-body and peer support intervention development to promote whole person health in patients with critical illnesses and their family members. The overarching scientific goal is to lay the foundation for the development of a mind-body peer support intervention for caregivers to patients with severe acute brain injury (SABI) by conducting preliminary quantitative and qualitative work and adapting existing interventions with stakeholder-engaged methods. Family caregivers to patients with SABI are at risk for chronic emotional distress, yet no tailored, efficacious interventions exist to promote their mental and physical wellbeing. Prior research suggests that mind-body skills (e.g., mindfulness, coping) can prevent chronic distress and promote wellbeing in caregivers in line with a biopsychosocial model of health. A peer-to-peer support model is an ideal approach to deliver mind-body skills to SABI caregivers for 3 key reasons: 1) peer support interventions capitalize on the value of social support to buffer against stress in caregivers, consistent with the Stress Buffering Hypothesis; 2) peer support interventions can powerfully spur health behavior change as explained by Dynamic Social Impact Theory; and 3) peer support interventions are more cost-effective, scalable, and appropriate for lower-resource settings than clinician-delivered interventions. An understanding of the support needs and perceptions of peer support among SABI caregivers is needed to inform the adaptation of a tailored mind-body peer support intervention. Informed by the intervention mapping framework, this study aims to 1) Investigate the association between availability of social support (Aim 1a) and satisfaction with social support (Aim 1b) and emotional distress over time in SABI caregivers using existing co-sponsor datasets (quantitative analyses; Aim 1); 2) Explore SABI caregiver perspectives on a mind-body peer support intervention through individual interviews (qualitative interviews; Aim 2); and 3) Engage SABI caregivers in facilitated workshop to co-develop the mind-body peer support intervention manual (stakeholder-engaged intervention design; Aim 3). Through these aims, the applicant will receive training in mind-body and peer support interventions, quantitative methods, advanced qualitative methods, and stakeholder engagement methods. The applicant’s training goals are supported by dedicated sponsors and collaborators with complementary expertise. This F31 will equip the applicant with the necessary training and preliminary data for future work towards the goal of supporting SABI caregivers, including a K23 to refine the proposed intervention and evaluate feasibility in an open pilot with exit interviews and randomized controlled trial. This F31 aligns with NCCIH’s strategic plan, including focus on the whole person and the integration of complementary and conventional care in real-world settings, and research on health promotion, resilience, and disease prevention. If successful, the proposed mind-body peer support intervention could be adapted for other populations of family members in critical care settings, with wide-reaching potential to be implemented in critical care settings and improve the long-term wellbeing of family members after critical illness.

NIH Research Projects · FY 2026 · 2024-01

Project Summary Inappropriate accumulation of calcium phosphate salts in cardiac valve and vasculature, also known as vascular calcification (VC), is increased with age, specific genetic disorders, coronary artery disease, valve disease, diabetes and chronic kidney disease. Calcification of these normally compliant organs has severe impacts on hemodynamics and cardiovascular function due to increased stiffness and/or tendency to rupture. There are currently no pharmacological therapies approved to directly prevent or treat VC despite the recognized deleterious effects of calcification in blood vessels and valves. Over the last 20 years paradigm-shifting studies have revealed some of the key genetic pathways, cell types and pathobiological processes that control the initiation and progression of VC These studies indicate that rather than a degenerative, untreatable inevitability, VC is a regulated and potentially modifiable process. The challenge is to understand which pathobiological processes predominate under specific disease conditions, and the critical upstream and downstream signaling pathways regulating them in order to identify biomarkers and target for therapeutic development. The overall goal of my research is to generate new scientific knowledge regarding the pathobiological mechanisms at play in diabetes and chronic kidney disease, highly prevalent diseases with extremely high risk for VC and its complications. We will continue to use a human first approach to identify key risk factors that may play a causative role in VC, and mechanistic studies in animal models, organoids, and cell culture experiments to identify key signaling pathways involved. The proposed studies will facilitate identification of biomarkers, therapeutic targets and therapies to treat this silent killer and improve human health.

NIH Research Projects · FY 2026 · 2024-01

Project Summary I am interested in the molecular biology and genetics of microbial social interactions. We use the saprophyte and opportunistic human pathogen Pseudomonas aeruginosa as a model to study social behaviors. P. aeruginosa uses a cell-cell signaling system called quorum sensing (QS) to coordinate gene expression. Because gene expression is coordinated at the population level, QS in P. aeruginosa, and other bacteria, is a means by which these single-celled organisms can cooperate and engage in specific group activities. In our prior NIGMS funded work, we established an experimental evolution system in which we demonstrated that P. aeruginosa QS can circuits diversify in response to changing environments. We also developed microbial co-culture systems, which allow us to ask about interspecies competition and how QS might impact interspecies interactions. The thrusts in this application ask 1) in cooperating populations of bacteria, what is the genetic basis for the diversification of the QS circuity, and does this diversification result in a division of labor where different individuals contribute separate functions to the group? and 2) how do the interactions between P. aeruginosa and other microbes influence P. aeruginosa behaviors, and how does QS contribute to these interactions? Bacteria, because of their rapid generation time and large population sizes, allow us to ask questions about social behavior that are not possible in organisms with smaller populations or longer generation times. The questions asked in this proposal will advance our knowledge of P. aeruginosa QS, cooperative behaviors in populations and interspecies interactions, and will inform efforts to target P. aeruginosa QS as a therapeutic approach.

NIH Research Projects · FY 2026 · 2024-01

ABSTRACT My lab has focused on developing methods to understand the causes and consequences of biological heterogeneity. Examples of heterogeneity include differences between the traits of individuals in a population, between cells in an organism or between proteins in a cell. Heterogeneity defines the phenotypic landscape at the organismal, cellular and molecular levels, shaping how biological systems respond to short-term perturbations and how species and tumors evolve. Heterogeneity can be encoded by genetic variants or it can arise from environmental perturbations or stochastically. My lab’s past successes include a plethora of new approaches to understand phenotypic heterogeneity arising from genetic variation. In particular, I developed deep mutational scanning, which can measure the consequences of tens or hundreds of thousands of genetic variants on different aspects of protein or cell function. Using deep mutational scanning, my lab has gained fundamental insights into protein structure and function, enhanced computational variant effect prediction and reshaped how human genetic variants are interpreted. Moreover, hundreds of labs around the world have taken up my methods and, collectively, the methods have been used to measure the effects of millions of genetic variants in hundreds of genes. Now, I am investigating the causes and consequences of non-genetic biological heterogeneity. For example, I am fascinated by questions like “how do isogenic cells in a population, each with a different transcriptomic, proteomic and metabolomic state, respond differently to a perturbation? How does the initial state of a cell determine its eventual fate? What, if anything, does cellular morphologic heterogeneity mean either in terms of cell state or fate?” However, answering these questions requires developing new methods. Thus, in this MIRA application, I am seeking funding to extend my work probing the basic biology of non-genetic cellular heterogeneity by developing a slate of new methods and using them to answer fundamental questions like those highlighted above.

NIH Research Projects · FY 2026 · 2024-01

ABSTRACT This implementation science R01 study aims to test a scale-up package to expand the delivery of pre-exposure prophylaxis (PrEP) for pregnant and postpartum women in Kenya. The pregnancy and postpartum periods are high risk for HIV acquisition. PrEP is an effective, woman-controlled, evidence-based intervention that is recommended during pregnancy in high risk regions in both World Health Organization and Kenyan guidelines. Delivery of PrEP integrated into maternal and child health (MCH) clinics is feasible, acceptable, and preferable to pregnant and postpartum women. Our team has led 3 implementation science (PIs: John-Stewart, Baeten, Kinuthia, Wagner) studies with >55 facilities focused on delivering PrEP integrated into MCH, testing implementation strategies to optimize delivery in western Kenyan counties. These represented the first and largest projects to deliver PrEP integrated into MCH. Integrated PrEP delivery is now ready for scale up more broadly, which will require new implementation strategies to target scale up. The proposed R01 will leverage the experiences of health care workers (HCW), expertise from research team members, and heterogeneity between sites and counties, to develop and test a package of scale-up strategies for improving PrEP delivery. This project’s aims are to: 1) develop and determine the impact of a Community of Practice (CoP) on stakeholder and health care worker engagement and knowledge, 2) determine the impact of a 3 component scale up package – CoP, quality improvement, and training & delivery toolkit – on implementation outcomes and PrEP clinical outcomes, and 3) identify combinations of facility- and regional-level attributes and processes that explain patterns of implementation. These research aims follow directly from Dr. Wagner’s K01 on which Dr. Kinuthia is a mentorship member. The joint leadership between the University of Washington and Kenyatta National Hospital provide a superb research environment for implementation science projects, with a strong >35 year history of collaboration between the institutions. The established investigative team includes expertise in implementation science, PrEP, HIV prevention during pregnancy and postpartum, engagement with Ministry of Health, mixed-methods, configurational analyses, and strong expertise in delivery of PrEP in MCH.

NIH Research Projects · FY 2026 · 2024-01

Investigating the Impact of Epigenetic Heterogeneity on Hematopoiesis Hematopoietic stem cells (HSCs) generate all blood and immune cell types of the body, and proper control of their self-renewal and differentiation is essential for human health.1,2 Since the work of Till and McCulloch, heterogeneity has been recognized to be a cardinal feature of HSC biology. Clonally related cells can differ in their lineage decisions and showed preference towards certain cell fates, even in identical environments.1–6 Researchers termed these skewed decisions as “lineage bias”. Epigenetic mechanisms, such as post- translational histone modifications (`histone marks'), are postulated to be a possible factor contributing to individual HSCs heterogeneity and its lineage bias. For example, recent studies suggest that levels of repressive histone marks at silent gene loci can vary considerably from cell-to-cell and repressive histone marks such as H3K27me3 can be stably inherited through generations. Since lineage-specifying genes involved in hematopoiesis are typically held in a repressive state until differentiation, I hypothesize that individual HSCs show substantial epigenetic state heterogeneity at lineage-specifying gene loci and that this epigenetic heterogeneity creates stable, heritable lineage biases in these cells. To test this hypothesis, I will determine the extent of epigenetic heterogeneity present in HSCs, as well as whether it impacts lineage potential for HSCs. Both of these questions remain unclear due to the lack of methods that can measure epigenetic states consisting of an array of histone marks and their dynamics within single cells. Recently, the Vaughan and Kueh group developed a methodology termed Single-Cell Evaluation of Post Translational Epigenetic encoding (SCEPTRE), which uses super resolution microscopy to resolve and quantify multiple histone marks at specific gene loci of interest simultaneously. I will use SCEPTRE to investigate the extent of epigenetic heterogeneity present in HSCs and its descendants (Aim1) first. Then, I will interrogate the heritability and impact of repressive epigenetic states by coupling SCEPTRE with live-cell tracking (Aim2). If successful, the project will provide an exciting first look into epigenetic regulation in HSCs at the single-cell level, paving the way for downstream studies to reveal fundamental aspects of their control and action.

NIH Research Projects · FY 2026 · 2024-01

Project Summary. Direct exposure to sunlight subjects cells to potentially damaging solar radiation. For superficial tissues such as the cornea, retina, and skin this is unavoidable. Biological circadian rhythms have evolved as a way to anticipate sunlight: by synchronizing cellular and behavioral rhythms to the 24-hour solar cycle, or “photoentrainment”. Most tissues in mammals contain an autonomous molecular circadian clock, and many of these are synchronized by the brain's central clock, the suprachiasmatic nucleus (SCN). However, we have found that some tissues located near the body's surface, such as skin and retina, have an ability to directly synchronize their local circadian clocks to short-wavelength light using the opsin, Opn5. In the case of retina and skin clocks, rhythmic environmental light dominates systemic circadian synchronization cues. The central hypothesis of this work is that opsins in cells typically exposed to sunlight respond to short-wavelength light, causing the release of diffusible signals which synchronize adjacent circadian clocks. This allows tissues to maintain a phase relationship with environmental light regardless of the phase of the animal's behavior. The specific aims of this proposal are to identify the intracellular molecular mechanism Opn5 uses to signal the presence of light using skin and retina as model circadian systems (Aim 1), identify the nature of diffusible signaling by which Opn5-cells influence adjacent cells and tissues (Aim 2), and determine the physiological significance of Opn5-mediated circadian clock synchronization in vivo (Aim 3). Our approach will be to focus on skin as a model circadian tissue due to its physiologic significance, its natural exposure to light, and the relative ease of distinguishing Opn5- specific photoactivation. However, studies will be repeated in retina for analyses on the similarity of signaling mechanisms in another photoentrainable tissue. The studies will employ a mixture of in vivo, ex vivo (organotypic tissue explants), and cell culture methods. The innovation of this project lies in the measurement of direct responses of clocks within mammalian tissues to environmental light and the mechanisms opsins employ outside of the visual system. The improvements of this proposal over past work are the 1) assessment of Opn5 signaling cascades for which there is still a gap in published literature, 2) assessment of tissue-level signaling dynamics and 3) a focus on the physiologic impacts of lighting environment on tissue health.

NIH Research Projects · FY 2026 · 2023-12

Project Summary The exact nature of how positive and negative affective stimuli reciprocally interact within the brain to influence learning and memory is not well resolved. Understanding how positive affective states impact discriminatory threat learning, memory retention, and extinction has broad implications for the effectiveness of cognitive behavioral therapies. Based on its connectivity to brain regions involved in threat responding, threat generalization, the negative affect of pain, and salience detection, we hypothesize that the central nucleus of the amygdala (CeA) is a key site for the integration positive and negative affective state information that is important for the regulation of threat learning and the influences of affective state. Our preliminary results demonstrate that co-presentation of a conditioned stimulus associated with positive valence with a conditioned stimulus associated with a negative valence can prevent, reverse, or facilitate the extinction of a generalized threat memory. We further show that this effect is dependent on the activation of neurons in the ventral tegmental area and the modification of fear encoding neurons of the CeA by dopamine release. In this proposal, we seek to resolve how positive stimuli influence fear encoding and threat discrimination at the level of activation of specific neurotransmitter receptors in the CeA to facilitate discriminatory threat memories. We further seek to resolve how positive and negative affective stimuli are differentially encoded in the CeA and how these cell types can influence each other’s activity during positive and negative valence learning. Resolving the basic brain mechanisms that allow positive affective stimuli to influence threat memories has important implications for improving cognitive behavioral therapies in the treatment of generalized anxiety disorders.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY Microglia, the primary mediators of innate immune activation in the brain, are increasingly recognized as key modulators of neuronal activity and excitability. There is growing evidence in many neurological diseases, including traumatic brain injury (TBI), that prolonged activation of the innate immune system can impede repair and promote disease, and it is not understood if or how microglia's impact on neuronal excitability might contribute or protect. One interesting microglial subtype that may be critical in the monitoring and feedback of neuronal excitability is the perineuronal satellite microglia. These microglia are juxtaposed to neurons with their soma and processes entwined around the neuronal cell body. Our published and preliminary data in TBI show a dramatic increase in the percentage of neurons with satellite microglia at both one week15, and several months after TBI that is associated with network hyperexcitability and behavioral dysfunction with deficits in reversal learning. However, our preliminary data indicate that satellite microglia suppress neuronal excitability, in control mice, but lose this ability in chronic TBI with an associated decrease in expression of P2Y12 receptors. With this proposal, we will investigate the role of perineuronal satellite microglia and P2Y12 receptors in neuronal, network and cognitive dysfunction after TBI. In aim 1, we will utilize transgenic mice that allow identification of microglia and associated neuronal subtypes to establish cell type-specific interactions of satellite microglia with neurons and the associated effect on neuronal function. In aim 2, we will test the mechanistic role of P2Y12 signaling in satellite microglial-associated neuronal interaction and function, network hyperexcitability and cognitive deficits after TBI. In aim 3, we will validate satellite microglial-neuronal interactions, P2Y12R expression, function and microglial expression profiles in human tissue in the context of TBI compared to our murine model. Results from these studies will yield an in-depth understanding of how microglial-neuronal interactions contribute to changes in neuronal excitability after injury and will give insight into therapeutic targets for TBI-induced circuit and cognitive dysfunction.

NIH Research Projects · FY 2026 · 2023-12

Cardiovascular disease (CVD) deaths in the United States approached 1 million in 2019, accounting for 33% of annual mortality. Americans consume 3,400 mg of sodium per day, nearly 50% higher than the limit recommended by the U.S. FDA and WHO. Population sodium reduction is a strategy to reduce CVD deaths. The WHO recommends four ‘best buys’ for sodium reduction: product reformulation, front-of-package labeling, institutional procurement, and public educational campaigns. The FDA set a goal in 2016 to reduce sodium consumption to 2,300 mg per day within ten years. Unfortunately, a lag exists in sodium policy implementation in the U.S. An urgent need exists to apply implementation science to reduce excessive sodium consumption to ultimately reduce CVD deaths. My research responds to this need by applying the RE-AIM framework to policy implementation in systematically assessing sodium policy reach, effectiveness, adoption, implementation, and maintenance. My preliminary studies in global CVD modeling and U.S. multisectoral policy implementation serve as a critical foundation in formulating the overall objective for the proposed research and in establishing study feasibility of carrying out this research. The rationale for this research is to provide policy makers with the information and tools they need to improve cardiovascular health. This study will analyze historical state-level data to estimate what ‘has been’ adopted, model to learn what ‘might be’ the health gain and its distribution in nationwide implementation, and conduct qualitative study about what ‘could be’ implemented. I analyze retrospective data on sodium policy adoption, maintenance, and reach in the United States from 2000 to 2022 (Aim 1). I model how many lives might be saved by implementing institutional procurement policy in all states in ten years (Aim 2). I assess capacity and constraints in multilevel implementation system and stakeholder’s conventional and active roles in qualitative study (Aim 3). Outcomes of this study include: (a) scientific evidence about current sodium policy implementation RE-AIM measures; (b) an interactive tool, Policy RE-AIM, to present the findings from the three Aims in lay language and through visualization; and (c) preliminary data to apply for future funding (e.g., an R01 award) to advance CVD policy modeling methods and to improve sodium policy implementation at the county level. The proposed research illustrates a novel application of RE-AIM to policy implementation science. It is innovative in developing an interactive tool that can be used in the future as part of a strategy to improve policy implementation to ultimately eliminate preventable cardiovascular health disparities. The intended impact of my study is to contribute to the application of policy IS to CVD prevention and accelerate the translation of cardiovascular science into real-world improvements in health equity. I have multidisciplinary mentorship with expertise in implementation science (Dr. Bryan Weiner), policy implementation (Dr. Jodi Sandfort), modeling (Dr. Yan Li), cardiovascular health (Dr. Chris Longenecker), as well as sodium and health (Dr. David Watkins).

NIH Research Projects · FY 2026 · 2023-12

In 2017, decompensated cirrhosis caused more than 1.32 million deaths globally and 62,943 deaths in the USA. A transjugular intrahepatic portosystemic shunt (TIPS), an intraparenchymal artificial shunt created between the hepatic vein (HV) and the portal vein (PV), is the most effective life-saving procedure to stop acute variceal bleeding (AVB) that cannot be controlled endoscopically. TIPS is also effective to decrease the risk of recurrence of tense ascites and improve renal dysfunction in patients with chronic liver disease. However, Intrahepatic puncture for TIPS creation remains one of the most challenging and radiation-intensive procedure, carrying high risks of various complications in up to 20% of cases even in the hands of experienced Interventional Radiologists. Intraperitoneal hemorrhage caused by liver and extrahepatic portal vein rupture is the most catastrophic major complication of TIPS creation. In children, TIPS has been employed to alleviate symptoms of refractory ascites and variceal bleeding, but only adult TIPS puncture sets are available for kids with portal hypertension. The intrahepatic puncture devices, complications and technical failure rate have not changed since TIPS was invented by interventional radiologists (IR). To address the critical issues described above, we are developing a novel in-plane IVUS/fiber optic imaging-integrated micro puncture balloon catheter system for TIPS creation. Tests of the fine needle/balloon catheter system in pig cadavers for TIPS creation resulted in 100% success of placing a 5Fr. catheter in the portal vein from the right hepatic vein. This novel fine needle(22G)/balloon catheter system is expected to eliminate the risk and significantly lower the difficulties of TIPS techniques, which will ultimately increase the availability of TIPS treatment. The in-plane IVUS and/or fiber optic imaging guidance will further facilitate TIPS creation. Therefore, we need to implement a series of experiments to bridge the gap between our laboratory innovation of a novel system for TIPS creation and clinical translation for treating patients with portal hypertension. At first, we will optimize the key components and parameters of the needle/balloon catheter systems using human excised cirrhotic liver specimens. Secondly, we will develop the technique of using fiber optic imaging and/or in-plane IVUS- integrated needle/balloon catheter system for TIPS creation in normal adult pigs, optimize the technical parameters, and further validate the technique of using the system for TIPS in pigs with liver cirrhosis and portal hypertension. At last, we will develop the technique of using this novel system for TIPS creation in baby and young pigs, optimize the technical parameters, and further validate the technique of using the system for TIPS in young pigs with portal hypertension. We are confident that the success of this project will lead to significant breakthroughs in the procedure of TIPS creation and benefit millions of patients with decompensated cirrhosis.

NIH Research Projects · FY 2025 · 2023-12

Project Summary This project aims to reveal the molecular mechanisms underpinning initiation of innate immune activation and the innate immune response during flavivirus infection. Flaviviruses comprise a number of emerging and re- emerging pathogens with a global disease burden of over 400 million infections each year. West Nile virus (WNV) is an emerging neurotropic flavivirus that serves as a model flavivirus and is itself the number one arbovirus infection in the United States. Infection with WNV elicits a primary innate immune response through activation of the RIG-I-like receptor pattern recognition receptor signaling pathway when RIG-I detects viral pathogen associated molecular patterns (PAMP). However, the identity and features of viral RNA (vRNA) that drive this response as well as how sequestered vRNA is sensed by cytosolic RIG-I remain unknown. The proposed research aims to identify the species of and molecular motifs within WNV vRNA that drive a productive innate immune activation and response (Aim 1) and to pinpoint the intracellular sites of vRNA-RLR interaction in the infected cell to define the relationship between vRNA localization within/outside of viral replication compartments and innate immune activation (Aim 2). Our preliminary data has uncovered a role for single-stranded viral replication intermediates (negative-sense vRNA) in driving early innate immune activation. In Aim 1, cellular subpopulations will be sorted based on innate immune activation status then subjected to RNA sequencing to further identify specific species of vRNA associated with innate immune activation. Further analysis of vRNA bound to RIG-I during infection will be performed with immunoprecipitation followed by RNA sequencing and specific immunostimulatory nucleotides will be mapped via individual nucleotide resolution cross-linked immunoprecipitation (iCLIP). In Aim 2, novel vRNA-RLR proximity ligation assays alongside immunofluorescence will be used to pinpoint cellular locations of innate immune activation. Innovative cryo- electron tomography (cryo-ET) coupled with RNA fluorescent in situ hybridization via correlative light and electron microscopy (CLEM) will be used to define the relationship between immunostimulatory RNAs and viral replication compartments at the ultrastructural level. In addition to revealing the identify and location of immunostimulatory vRNAs during WNV infection, the work in this proposal will facilitate training in multiple cutting-edge techniques including iCLIP, RNA sequencing and bioinformatics, and high-resolution imaging including cryo-ET and CLEM, with the support of co-sponsors who are experts in flavivirology and innate immunity and viral ultrastructure and imaging techniques. Technical and conceptual training provided will support the transition of the trainee to an independent investigator in RNA virology and innate immune activation, while research findings will provide conceptual advances in our understanding of RLR-signaling during flavivirus infection.

NIH Research Projects · FY 2026 · 2023-11

Vaccination is one of the most effective tools to control the spread of infections and reduce child mortality, however the WHO estimates 1.5 million children under 5 years old die each year from vaccine preventable illnesses. Vaccine efficacy may be reduced by host immune modulating factors that impair optimal immune responses, including exposure to HIV, infection with cytomegalovirus (CMV), and other factors related to maternal or infant infections and inflammation. Regions with high prevalence of HIV, poverty, and malnutrition also often have sub-optimal vaccine coverage. Together, these factors put large populations of infants and young children at risk for preventable deaths. Several studies have reported deficits in antibody responses to routine childhood vaccines in HIV-exposed uninfected (HEU) compared to HIV-unexposed (HUU) children, but few studies have looked at cellular vaccine responses in HEU. There is also evidence that infection with CMV, which elicits profound immune activation and inflammation following infection, impacts vaccine responses, but results are not conclusive. Thus, substantial gaps remain in our understanding how exposure to maternal HIV infection and infant infection with CMV may impact humoral and cellular vaccine immune responses. Our overarching hypothesis is that early-life exposure to immune modulating infections, specifically HIV and CMV, may affect immune responses to vaccines given during infancy as well as to vaccines first administered in middle childhood. We will extend follow-up of an existing birth cohort of HEU and HUU Kenyan children previously followed from pregnancy to 4 years postpartum with longitudinal collection of specimens and clinical data, including vaccine records. We will determine the effect of HIV exposure and/or early CMV acquisition on humoral and cellular responses to infant vaccines from birth through 4 years of age (Aim 1), and on humoral and cellular responses to influenza vaccination in middle childhood at 6 years (Aim 2). Data from our existing project (virome/microbiome, enteric enteropathy, growth, child and maternal in utero inflammation, infections) will be assessed as modulators of vaccine-induced immunity in HEU and HUU children from infancy through toddlerhood, early and middle childhood (Aim 3). The proposed studies offer an unprecedented opportunity to understand how HIV, CMV, malnutrition and inflammation alter early life and long-term immune reactivity in African children. Results from this study have great potential to inform novel vaccination strategies and interventions for children in areas of high HIV prevalence, CMV and malnutrition.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Over a third of American adults engage in unhealthy substance use (USU), which includes a spectrum ranging from any illicit drug use or alcohol consumption above guideline-recommended levels through more severe substance use disorder. USU results in increased mortality through overdose and substantial morbidity including reduced functioning, poorer chronic disease outcomes and increased health care utilization. A broad range of biological, psychological and social factors contribute to USU. However, while primary care clinicians frequently encounter patients with USU, they rarely screen for, diagnose, or treat USU using a comprehensive approach that would account for all these factors. Furthermore, existing strategies and models to improve USU care frequently fail to consider practice and community context, and thus have not been widely disseminated. In our study, Supporting Unhealthy Substance use care Through a whole person Approach and user centered INtegration into primary care (SUSTAIN), we propose to engage primary care practices and individuals with lived USU experience in codesigning a whole person USU intervention and collaboratively test the implementation of the SUSTAIN intervention in primary care practices. In phase 1 of our study, we will convene a series of stakeholder groups with practice representatives and individuals with USU who will codesign a change package that includes a menu of interventions that considers the psychological, social, and biological factors that contribute to USU. In phase 2, we will conduct a cluster randomized controlled trial with waitlist control in 24 practices with 50 patients each from 2 practice-based research networks, the WWAMI (Washington, Wyoming, Alaska, Montana and Idaho) region Practice and Research Network and the Virginia Ambulatory Care Outcomes Research Network. Participating practices will join a learning collaborative where they will be supported in implementing their customized approach to identifying and treating USU by selecting from the menu of interventions developed in Phase 1. The primary outcome will be patient functioning (measured by the PROMIS-29-v2) and secondary outcomes will include unhealthy substance use (TAPS-1), recovery (RAS), health care utilization and process outcomes. Guided by the Consolidated Framework for Implementation Research (CFIR), we will evaluate implementation outcomes and conduct qualitative interviews with a subset of patients and clinicians. Our proposed intervention will enhance equity since the intervention is adaptable to the needs and preferences of each practice and individuals served by the practice. It is innovative in that it transcends the current focus on implementing a specific model and recognizes and adapts to the diversity of primary care practices in communities served, organizational structures and payment models. If successful, the SUSTAIN model can transform how primary care practices care for individuals with USU in a sustainable and whole person approach, improving patient outcomes and enhancing communities.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Opioid abuse is devastating communities across the United States and is responsible for untold suffering. The synthetic opioid fentanyl, whether due to prescription or illicit use, is involved in nearly half of reported opioid- related deaths. Unlike heroin and morphine, fentanyl is commonly administered through non-intravenous routes, but we have a limited mechanistic understanding of fentanyl use and abuse vulnerability. A leading hypothesis for the transition from use to abuse, in vulnerable individuals, is the recruitment of drug-induced gene expression changes in certain brain circuits and cells following repeated drug exposure. However, identifying such circuit- and cell-type specific populations, in combination with their underlying genetic and functional adaptations, is often limited by the resolution, throughput, and data registration of current assays. The overall goal of this project is to (i) generate machine--guided behavioral characterization of oral and intravenous fentanyl self-administration and reinstatement in male and female mice that captures individual risk vulnerability, and (ii) perform brain-wide mapping by integrating new enabling technological platforms including single-cell spatial transcriptomics (Pixel- seq), whole brain cell type specific circuit connectivity, and functional distributed brain-wide neural activity recordings (Neuropixel 2.0), recently developed by PIs in our team. To realize the potential for the molecular and functional brain mapping and data registration, we have three Specific Aims: 1) perform single-cell spatial transcriptomic profiling, with circuit-specificity, in mice with varying degrees of fentanyl-seeking behavior; 2) perform single-cell whole-brain activity mapping, with cell-type and circuit-specificity, in mice with varying degrees of fentanyl-seeking behavior; 3) perform brain-wide distributed large-scale electrophysiological recordings during oral fentanyl-seeking and integrate spatially resolved transcriptomic data with recording data. In the first Aim, Pixel-seq will be used to generate cell atlases, analyze drug- and behavior-associated spatially conserved gene expression, and map neuronal connectivity by coupling with retrograde viral tracing. In the second Aim, we will perform immediate early gene-based whole-brain activity mapping, contextualized by cell type-specific afferent connectivity, after fentanyl self-administration and reinstatement. In the third Aim, we will first perform Neuropixel2.0 electrophysiological recording of the brain-wide distributed regions of interest, and then integrate and align Neuropixel2.0 and Pixel-seq data in a new assay called NeuroPixel-seq (NP-seq). The proposed project is innovative as our integrative approaches will, for the first time, generate spatial multimodal data of unprecedented depth and resolution within the context of opioid relapse risk. It is significant because our data will provide a much-needed accessible oral fentanyl self-administration model for the neuroscience community, paired with the first fentanyl cellular-resolution atlas, allowing non-specialized labs an accessible beachhead for participating in the identification of the mechanistic basis for fentanyl use and relapse in mice.

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract The proposed project seeks to expand the existing UW multidisciplinary long COVID clinic (the Post COVID Rehabilitation and Recovery clinic) to improve access and coordination of care for patients with long COVID. This project will allow the team to improve care delivery for all patients, but will emphasize improving access to multidisciplinary care for underserved patients including patients in rural communities, Alaska, and in Latinx, Native American, and SE Asian communities. With additional funding, we will make an enormous impact on a large region throughout WWAMI, focusing initially on Washington and Alaska, and Idaho, Montana, Wyoming in the later years. The aims of this proposed project include Aim 1: Improve clinical care delivery and expand access to the UW PCRRC. The goal of this aim is to improve access and care delivery for patients from Washington and Alaska that are referred to the Post-COVID Rehabilitation and Recovery clinic. The key areas that we will be expanding in this project include: 1) coordination of care; 2) behavioral health support; 3) vocational services; and 4) electronic consults. Aim 2: Engage underserved communities to enhance long COVID education and connection to local resources. The overall goal of our community engagement activities will be to foster authentic longitudinal relationships, promote ongoing collaborations and address the needs of community members. We will accomplish this by: 1) forming a community advisory board to guide our engagement activities; 2) producing publicly accessible educational webinars (community conversations) addressing COVID and long COVID; 3) producing culturally responsive patient education materials on long COVID in English, Spanish, and SE Asian languages; and 4) participating in community events including town hall meetings, community health fairs, pop-up vaccination events to share information about the UW long COVID clinic and long COVID. Aim 3: Improve patient access in the WWAMI region by training a network of clinicians specializing in long COVID care. In this aim, we will provide ongoing education and support to community PCPs, psychologists, and physical therapists with a focus on providers in under-resourced areas across the WWAMI region, including rural areas. We will accomplish this by: 1) In partnership with the Washington State DOH, we will create an advanced training certificate program for long COVID. 2) We will conduct monthly interactive case-based webinars. 3) We will also identify local physician and physical therapy leads who can be trained to be local experts in the care of patients with long COVID and are a part of our extended UW PCRRC network. 4) We will hold weekly virtual office hours that are pre-specified with a scheduling system to allow community providers to discuss cases and ask questions that will help them with their own management of patients with long COVID.