University Of Washington

NIH Research Projects · FY 2026 · 2022-01

Project summary Each year over 100,000 Americans begin dialysis when their kidneys lose the capacity to remove toxins from the blood and half of them die within five years. These high mortality rates are in part due to the ineffectiveness of dialysis at removing protein-bound uremic toxins (PBUTs) such as indoxyl sulfate and p-cresol sulfate, which would otherwise be removed by healthy kidneys. Herein, we propose a non-invasive ingestible bubble tea to reduce the blood concentration of PBUTs and improve the outcomes of patients with end stage kidney disease (ESKD). In contrast to conventional treatments, which attempt to remove toxins from the blood by dialysis, this research offers a radically different approach, which prevents the formation of toxins at their origin. PBUTs are byproducts of gut microbial metabolism; indole and p-cresol originate in the gut from fermentation of amino acids and both solutes are sulfonated in the liver to form the PBUTs indoxyl sulfate and p-cresol sulfate. We propose a novel microbe enriched bubble tea which will expand the capacity of the gut to degrade the uremic toxin (UT) precursors (indole and p-cresol) and thereby prevent their sulfonation to toxic forms in the liver. The bubble components will be engineered to remain tight in the stomach, hence protecting the hydrogel entrapped microbes from low pH, and then swell at neutral pH in the small intestine and colon, allowing UTs to diffuse into the hydrogel where the UT degrading microorganisms will deactivate them. Finally, the bubble tea will be excreted intact without altering the natural gut microbiome. This concept builds on the successful applications of microbe enriched hydrogels in bioengineering and environmental engineering for drug delivery and wastewater treatment. We will develop the bubble tea by first enriching novel indole and p-cresol degrading microbes from the environment, second engineering specialized hydrogel carriers for the microbes, and finally replicating their trip through the gut in well controlled bioreactor units. The substrate profiles, pH gradients, and removal rates will be experimentally measured and mathematically simulated to visualize spatial localization of bacteria and to determine UT degradation over time. Delivering these hydrogels in a formulated bubble tea drink could improve the lives of chronic kidney disease and ESKD patients by reducing the burden from uremic symptoms and complications.

NIH Research Projects · FY 2026 · 2021-12

ABSTRACT There is a significant need for clinically trained practitioners with structured research training in environmental health to translate research findings to clinical and policy realms. The overarching goal of the University of Washington Pediatric and Reproductive Environmental Health Scholars (UW PREHS) career development program is to bridge clinical training to research independence, through a mentored research experience in pediatric and reproductive environmental health. The program integrates the outstanding research and training opportunities at the University of Washington, forming a novel partnership among the Departments of Pediatrics, Family Medicine, Obstetrics/Gynecology, Environmental & Occupational Health Sciences, and Epidemiology. Nationally recognized PREH physicians-scientists and Pediatric Environmental Health Specialty Unit (PEHSU) leaders, Dr. Catherine Karr and Dr. Sheela Sathyanarayana, are uniquely suited to lead the program to prepare future leaders in PREH research and research to policy and practice translation. Our primary aims are to: 1. Recruit and train scholars in PREH science, rigorous research methodology and research skills. 2. Support scholars in completion of an impactful PREH research project. 3. Involve scholars in research translation practice activities through the NW Pediatric Environmental Health Specialty Unit. 4. Establish and maintain a collaborative network of mentors for scholars to support their success in the UW PREHS program and in transition to independence upon completion of the program. Scholars will participate in a tailored career development program anchored in a set of core competencies spanning four primary domains: 1) PREH content expertise, 2) Research methods development, 3) Practice based research translation (PEHSU), and 4) Career development toward independence. Central structural elements include mentor advised individualized learning plans, a novel PREH seminar series, involvement in PEHSU consultation and outreach, and development and implementation of at least one research project. Ongoing evaluation plans for scholars, mentors, and the program provides continual improvement for this new program. A senior Advisory Committee will provide oversight of the program evaluation based on identified benchmarks. The program is poised to recruit and train up to 6 scholars for 2-3 year periods each. Scholars will be prepared to serve not only as scientists, but clinician-advocates and trusted and impactful voices on policy and practices to improve environmental health conditions of children and families.

NIH Research Projects · FY 2026 · 2021-12

ABSTRACT Management of patients with metastatic non-small cell lung cancer (NSCLC) requires navigation of an increasingly diverse therapeutic landscape. Although immune checkpoint inhibitors (ICI) of anti-programmed cell death 1 (PD1) and its ligand PDL1, in combination with chemotherapy (chemoICI), are standard of care for metastatic NSCLC and have improved survival in some patients, the majority are subject to treatment-related toxicity at significant financial burden with little clinical benefit. Radiation therapy can prolong survival in patients with limited sites of metastatic disease (oligometastatic), or limited sites of progressive disease (oligoprogression) on systemic therapy, but no consensus exists on which patients and lesions would benefit from irradiation. Patient selection and treatment adaptation through early response assessment is an unmet need to increase the effective combination of chemotherapy, immunotherapy, and radiation therapy in metastatic NSCLC and improve outcomes. Biomarkers are critical to our understanding of complex response patterns to chemoICI and radiation. In patients with newly diagnosed metastatic NSCLC starting chemoICI per standard of care, we propose to assess and monitor treatment response by combining positron emission tomography (PET) imaging of macroscopic disease burden and circulating immunologic biomarkers of occult systemic disease burden in support of precision therapy through the following aims: (1) construct clinical PET imaging and circulating immunologic biomarker signatures of chemoICI response patterns to risk stratify patients into (a) early widespread progression, (b) oligoprogression, and (c) responsive disease; (2) construct clinical PET imaging and circulating immunologic biomarker signatures of oligoprogressive radiation therapy response patterns to identify patients and lesions that benefit from ablative radiation; and (3) correlate localized clinical PET imaging and global circulating immunologic biomarkers with survival outcomes. Fluorodeoxyglucose (FDG) PET scans and peripheral blood draws will be performed prior to chemoICI, 3 weeks into chemoICI, and 12 weeks into chemoICI. For patients who develop oligoprogressive disease, we will acquire FDG PET scans and peripheral blood prior to and 1-month post radiation therapy. We will develop combined quantitative PET imaging and circulating immunologic biomarker signatures of chemoICI and radiation response that stratify patients into the following groups: (i) high-risk patients predicted to develop rapid widespread progressive disease who require aggressive second-line systemic therapy, (ii) moderate-risk patients predicted to develop oligoprogressive disease who require consolidation radiation to high-risk lesion targets, (iii) low-risk patients predicted to have durable long-term response to first-line therapy. Successful completion of this project will support the launch of a clinical trial on biomarker response-adaptive chemoICI and radiation therapy in patients with metastatic non-small cell lung cancer, in order to improve cancer control and survival.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Women have a higher lifetime risk of developing Alzheimer's disease (AD) than men. This increased risk is not fully explained by differences in longevity, hormones or brain structure. Women who carry at least one copy of the APOEɛ4 allele, the strongest genetic risk factor for late onset AD (LOAD), have accelerated neuropathology. However, some studies suggest a faster decline in men, suggesting that sex bias may differ depending on the stage of the disease. Here, we will investigate how the sex chromosome complement and sex-linked genes influences sex differences in onset and progression of LOAD. Genome-wide association studies have identified genetic and epigenetic risk factors for LOAD, but the sex-chromosomes are often excluded in these studies meaning there is a lack of data on sex-linked genes. Males have unique Y-linked genes and females have higher expression of genes that escape X inactivation. Interestingly, many of the escape genes are related to immune function and neuroinflammation is a hallmark of AD, suggesting that these genes may directly contribute to disease progression. To address the impact of sex-linked genes combined with APOEɛ4 alleles on neuroinflammation in LOAD we will use unique cellular models and AD tissue for leveraging integrated omics and functional studies. We will evaluate the functional roles of sex chromosomes and sex-linked genes in brain cell types using human induced pluripotent stem cell (hiPSC) models. We have derived isogenic pairs of hiPSCs with a different number of sex chromosomes on the same genetic background (XXY/XY or XXX/X). These new hiPSC lines minimize variability between individuals, as well as environmental or hormonal confounders. We will generate isogenic pairs of these lines with ɛ3/3 or ɛ3/4 alleles by gene editing. After differentiation of hiPSC into neurons, microglia, and brain organoids we will employ a combination of `omic' analyses and functional assays focusing on neuroinflammation and neurodegeneration. This approach will identify sex-linked candidate genes, which will be tested for dosage effects by knockdown and overexpression. These in vitro studies will be validated in human tissue collected by the Precision Neuropathology Core from our Alzheimer's Disease Research Center brain bank. Using pathologically characterized AD brains we will employ myeloid-specific single-nucleus RNA sequencing to determine the effects of sex and APOEɛ4 genotypes on microglial subtypes and neuroimmune gene expression. Our new team combines expertise in hiPSC modeling, sex-linked genes, neuroinflammation, `omic analyses and neuropathology. This integrative study will help understand sex-specific genetic factors and how those factors interact with APOEɛ4 risk to modulate cellular dysfunction and pathology, thus providing novel insights into how to tailor a more effective treatment for AD.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Significant transitions at midlife can be positive for some, but many also experience emotional challenges, physical health declines, and high stress in adapting to new social roles. Yet, health and functioning at midlife have received less attention compared to older ages because disease and disability remain relatively rare. This combination of growing health challenges and low rates of disability points to opportunities for intervention to prevent and delay more serious illness and impairment. This study addresses important gaps in knowledge to better capitalize on these opportunities. Little is known about how midlife may be different for those now in their 40s and 50s—often labeled Generation X—who are much more diverse demographically and in their family formation and education/career paths than prior generations. Most existing midlife research has also focused on relatively narrow health outcomes with little consideration of the interplay of physical and mental health and functioning in work and civic life. Recent studies showing significant health declines in new midlife cohorts additionally underscore the need for a focus on disparities and sociodemographic differences in midlife health and mechanisms to explain them, particularly increasing alcohol misuse and its central role in midlife health risks. This study brings strengths to examining these issues by drawing on existing longitudinal data from the Seattle Social Development Project (SSDP) and collecting new data in early midlife. SSDP has followed a gender-balanced, multiethnic, and socioeconomically diverse panel of 808 participants across 15 waves from age 10 to 39 with high retention. Data include theory-guided assessments of the social environment throughout; unique longitudinal assessments of neighborhood environments that integrate self-report, Census, and GIS measures; and longitudinal measures of alcohol and other substance use and disorder, mental and physical health, and functioning in work, community, and civic life. New data augment and extend existing measures into midlife and enable the examination of midlife-specific experiences. The study aims are to examine (1) how sociodemographic statuses and transitions among those now entering midlife influence comorbid health outcomes in early midlife; (2) how alcohol use, misuse and related behaviors during midlife influence midlife health, as well as the influence of life course trajectories of alcohol use and misuse leading up to midlife; (3) how malleable social and neighborhood environmental factors influence alcohol use and misuse over the life course and, in turn, affect midlife health; and to (4a) examine the long-term malleability of social environmental factors from early adolescence to midlife by testing the mechanisms of a childhood intervention (embedded in the existing study); and (4b) identify promising adult intervention targets by conducting focus groups with participants to help design and tailor prevention programs. With guidance from empirical findings and participants themselves, the goal of the study is to provide researchers and health practitioners with specific recommendations for preventive interventions addressing public health priorities for midlife adults.

NIH Research Projects · FY 2025 · 2021-09

Project Summary/Abstract The last few decades have seen major inroads into detailing the physiological mechanisms supporting vision as well as therapies aimed at rescue and repair of neurons affected by retinal diseases. For the continued evolution of treatments and their rapid translation to the clinic, it is essential to find a non-invasive, all-optical biomarker to monitor the efficacy of disease and potential therapeutic agents. To this end, we propose to develop the optoretinogram, or ORG, the optical analog to the electroretinogram (ERG) which is the current gold standard for retinal function assessment in humans. The ORG is rooted in classical interferometry and enables a highly sensitive assay of how neurons interact with light. Using this technique, our group has demonstrated the ability to visualize light-driven neural activity across a range of spatiotemporal resolution – from single cells to a collection of neurons, and from µsecs to ms timescales. Here, we aim to expand the capabilities of the ORG and demonstrate its efficacy for basic science and clinical applications. The proposed technology is built upon a solid foundation of established approaches, and combines them in new and complementary ways to achieve an optimal combination of speed, resolution and sensitivity geared towards overcoming the key challenges faced with imaging cellular structure-function in humans. The core technologies are phase-resolved OCT, an eye-safe, interferometric method to measure nm-scale changes at ms time scales in vivo, adaptive optics (AO) to overcome ocular aberrations, increase the signal-to-noise and allow resolution down to single cells and real-time eye tracking to overcome eye motion and allow targeting, recording and averaging of responses from single and a collection of retinal neurons. These are implemented across three ORG platforms. At the University of Washington, we will refine the line-scan phase-resolved OCT with improvements in optical design and eye-tracking and use it to characterize the basic properties of phototransduction and inner retinal function in healthy human volunteers and patients with retinal degenerations. At Stanford University, we will develop a similar line-scan system for rodents, and together with transgenic models and pharmacology, determine the biophysical mechanisms that underlie the ORG and develop templates for human recordings. At UC Berkeley, we will push the envelope of speed and sensitivity by incorporating a real-time eye-tracking system to drive an AO-OCT interferometric probe, with the aim to measure the tiniest and briefest neuronal changes in the human retina. This bioengineering research partnership will benefit from complementary expertise, research direction and ORG implementation across the three sites, and the use of common approaches for image/data analysis, eye tracking and visual stimulation. Ultimately, the aggregate technology will facilitate a deeper mechanistic understanding of early visual processing and eye disease, and provide entirely new avenues for accelerating therapeutic interventions.

NIH Research Projects · FY 2024 · 2021-09

ABSTRACT Comprehensive identification and characterization of senescent cells in morphologically intact human tissues is important for understanding senescence in vivo and the targeted removal of these cells to improve healthspan and lifespan. This task has been challenging due to the lack of universal and unequivocal markers characterizing the senescence state, which reflects the complexity of the senescence phenotype and the existence of highly heterogeneous senescence programs. A preferred avenue for discovering senescence markers is to spatially map ‘omics’ states of cell types in different tissues and life stages at single cell resolution. The overall goal of this project is to (i) develop a spatial, single-cell-resolution, multimodal method that simultaneously analyze transcriptome, open chromatin, and proteome (or secretome), and (ii) optimize and scale it for mapping senescent cells in human tissues. The PI’s laboratory has recently developed a novel technique PIXEL-seq (polony-indexed library-sequencing) and applied it to spatially profile transcriptome with 1-µm resolution and high RNA capture efficiency. To realize its potential for studying in vivo senescence mechanism and production-scale data generation, three specific aims will be pursued: 1) In UG3 Year 1, demonstrate PIXEL-seq-based spatial transcriptome, proteome, and ATAC-seq assays with single-cell resolution; 2) In UH3 Year 2, optimize and combine these assays for human tissue mapping; and 3) In UH3 Years 3-4, scale up application to human heart, liver, and lung tissue mapping. Under the first aim, PIXEL-seq will be developed to achieve single-cell resolution by image-guided cell segmentation (Aim 1A) and expanded to spatial proteome (Aim 1B) and open chromatin accessibility assays (Aim 1C) by rendering DNA-tagged antibodies and Tn5-treated chromosomal DNAs, respectively, to capture by polony gels. For the second aim, the proteome assay will be optimized and scaled to 200-plex using polyclonal mini-binders, allowing the cross-validation of senescence markers and associated isoforms and post-translational modifications (Aim 2A). These assays will be integrated for multimodal data capture and validated using human tissues (Aim 2B). In the third aim, the application will be scaled up by increasing throughput of polony gel fabrication (Aim 3A) and to deliver to the CODCC for public release of high- quality data on several sites of multiple organs from several individual tissue donors (Aim 3B and 3C). The investigators will also participate in the Consortium common project and other collaborations yet to be formed. The proposed project is innovative in that this method will for the first time generate the spatial multimodal human tissue data at unprecedented depth and resolution. It is significant because the assays do not require specialized equipment and can be widely implemented in the SenNet and other single cell consortia.

NIH Research Projects · FY 2024 · 2021-09

Abstract The goal of this project is to develop a clinical real-time spectroscopic photoacoustic/ultrasound (PAUS) system for molecular guidance of interventional procedures through a partnership between UW and GE Research. Recently, we proposed a new, fast-sweep concept for PA imaging. To put this concept into practice, we first developed a unique, compact, diode-pumped, tunable (700 -900 nm) laser operating at very high (up to 1000 Hz) repetition rates and relatively low (~ 1 mJ) pulse energies, and a fiber-optic delivery system to sequentially couple laser pulses into the imaging probe. In addition to US B-mode, and all other US modes, the system simultaneously produces real-time (50 Hz) spectroscopic PA images, which were combined for the first time for real-time PAUS imaging. A unique feature is automatic on-line laser-fluence compensation and motion correction, enabling quantitative optical absorption spectroscopy at every image pixel. Spectroscopy can identify substances opaque to US based on their molecular constituents (drugs/contrast agents), and quantify tissue functional changes (e.g., blood oxygenation and its concentration) within the image; in addition, manipulation with a needle is better visualized with PA. UW will work with GE Research to integrate spectroscopic PAUS into a high-end US scanner to create a clinical-grade PAUS system, and test whether it can improve interventional procedure guidance in general and, particularly, in ethanol (EA) ablation therapies of recurrent thyroid tumors. The prognosis for most people with thyroid cancer after primary treatment is very good, but the recurrence rate or persistence can be up to 30%. If recurrent cancer is confirmed, image-guided nonsurgical procedures such as EA or radio frequency ablation (RFA) are commonly used alternatives to more invasive procedures. Although US helps position EA and RFA needles, on-line imaging of the ablative area and confirmation of ablation remain difficult for US. When the recurrent nodule (especially the capillary network in it) is not entirely treated, the cancer will return with possible metastasis. We hypothesize here that real-time spectroscopic PAUS will improve the efficacy of ablation procedures and dramatically reduce procedure repetitions. If successful in this initial stage, the project will move to a clinical trial to both guide and validate ablative therapies and explore real-time spectroscopic PAUS for other interventional procedures. SA1 will integrate our unique laser and scanning fiber-optic delivery system with a clinical GE US scanner for real-time spectroscopic PAUS. Then, SA2 will develop real-time signal processing tools for motion correction and fluence compensation and imaging protocols for spectroscopic PAUS. SA3 will focus on optimizing the PAUS system using phantom and ex vivo studies. Finally, in SA4 the developed PAUS system will be used to test the clinical applicability of PAUS guidance with three in vivo models, including small animal studies of thyroid cancer, an animal model approximating human anatomy, and pilot measurements on human subjects.

NIH Research Projects · FY 2024 · 2021-09

PROGRAM ABSTRACT - Data Science for Child Health Now in Ghana (DS-CHANGE) Training Program Our goal for the DS-CHANGE Training Program is to build data science capacity at Kwame Nkrumah University of Science and Technology (KNUST) and develop a cadre of qualified data scientists focused on child health in Ghana. We will develop this cadre of scientists with mentored training and experiential learning that cross-sect biomedical data science disciplines (applied mathematics, computer science, clinical informatics, biostatistics, epidemiology), health conditions (malaria, injury and congenital anomalies), and biomedical domains (e.g. pediatrics, parasitology). Faculty and graduate student trainees will tackle computationally complex child health problems in Ghana. Our program focuses on 3 of the top 10 causes of child death/disability in Ghana including malaria, injury, and congenital malformations (orofacial clefts). We aim to: Aim 1: Deliver a comprehensive mentored interdisciplinary training program that cross-sects data science methodologies, health conditions, and biomedical domains to a diverse group of Ghanaian graduate-level MS and PhD trainees. Aim 2: Increase KNUST faculty and institutional capacity in biomedical data science by (a) facilitating cross-training in data science methodologies; (b) developing deeper expertise in biomedical data science methods; and (c) building teaching and mentoring proficiency in biomedical data science. Aim 3: Develop proficiency of faculty and graduate trainees in effective methods of team science so that interdisciplinary teams with minimal overlapping expertise can function synergistically. Collaboration: This program builds on established collaborations between KNUST and the University of Washington (UW), Seattle Children's Hospital and Research Institute, and the non-profit Smile Train. Approach: KNUST graduate trainees will obtain a Master or PhD degree in data science from KNUST that will be bolstered by a 3 month externship in Seattle with a thesis on a real world Ghana child health problem. KNUST faculty trainees will participate in faculty exchanges with UW faculty. Select KNUST faculty will complete a UW Professional Certificate Program in a data science domain. All trainees will participate in a monthly Zoom seminar to enhance interdisciplinary effectiveness. Impact: We will train 13 graduate trainees and 16 faculty for a robust biomedical data science graduate program at KNUST. Trainees will compete successfully for research funding, will contribute to the evidence base in child health, and will take up positions as leaders in data science and child health at KNUST and other Ghanaian institutions.

NIH Research Projects · FY 2025 · 2021-09

HIV incidence remains unacceptably high for adolescent girls and young women (AGYW). Pre-exposure prophylaxis (PrEP) HIV prevention tools are promising, with tenofovir (TFV)-based daily oral PrEP and the dapivirine vaginal ring (DPV-VR) recommended by WHO for cisgender women at-risk for HIV. Kenya is a leader for PrEP delivery in Africa and efforts are ongoing to increase PrEP access with AGYW as a priority group. Our team pioneered integrated PrEP delivery in family planning (FP) clinics in Kenya; yet, 40% of Kenyan women access contraception without interfacing with facilities, including at retail pharmacies, and would be missed by facility-based PrEP platforms. Retail pharmacies can increase options for reaching at-risk individuals with PrEP and efforts are underway to define pathways for pharmacy-delivered PrEP in Kenya. We adapted our FP clinic- based PrEP model and piloted PrEP delivery facilitated by nurse-navigators for AGYW seeking contraception at pharmacies. AGYW offered daily oral PrEP frequently initiated, planned to continue use, and were willing to pay for PrEP at pharmacies. Pill burden was a common reason for declining oral PrEP and could be addressed by offering DPV-VR. Through close collaboration with the Kenya Ministry of Health both, national- and county-level, we propose a cluster RCT at 20 pharmacies in Kisumu, Kenya–a region with an HIV prevalence of up to 28% among women–to test the effectiveness of utilizing nurse-navigators at retail pharmacies to enhance AGYW PrEP use. We will expand on our successful pilot to offer both daily oral PrEP and the DPV-VR and prospectively ascertain PrEP outcomes (initiation, persistence, adherence) among AGYW. This effectiveness-implementation hybrid RCT is designed to expedite translation into practice by evaluating clinical effectiveness alongside implementation and cost outcomes. We hypothesize that combining nurse-navigators with pharmacy-based PrEP will provide a cost-effective strategy for delivering novel HIV prevention tools to AGYW in HIV high-burden settings. Aim 1 will determine the effect of nurse-navigators on PrEP initiation, persistence, and adherence among AGYW seeking contraception within a pharmacy-based PrEP delivery model through a 2-arm cluster RCT among HIV-negative AGYW at 20 retail pharmacies in Kisumu, Kenya. AGYW seeking contraception will be offered PrEP and self-select daily oral PrEP or the DVR-VR. Primary outcomes will be proportion of AGYW accessing contraception that initiate PrEP, persist with use at 10 mos., and adhere (quantified by TFV or DPV hair levels). Secondary outcomes will include PrEP selection (PrEP pills vs. DPV-VR), STI incidence, and adherence cofactors. Aim 2 will identify potential barriers and facilitators to acceptability, feasibility, and client satisfaction of nurse-navigators and DPV-VR delivery for AGYW accessing PrEP at retail pharmacies through a qualitative evaluation guided by Proctor et al. Aim 3 will estimate the cost-effectiveness of implementing pharmacy-based PrEP delivery with nurse-navigators. This will be the first study aiming to improve PrEP access for AGYW using a pharmacy-based model and will prime pharmacies to deliver novel PrEP agents in the pipeline.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Cervical cancer (CC) is almost entirely preventable with current technologies, yet, it remains the most common cancer and the most common cause of cancer death among women in Eastern Africa. Globally, CC is the 4th most common cause of cancer incidence and mortality among women, and the leading cause of cancer in 42 low- and middle- income countries (LMICs), where 90% of CC deaths occur. To achieve the 2018 World Health Organization (WHO)'s call to action towards global CC elimination, there is an urgent need to adapt, implement, and scale-up effective technologies in LMICs. The mainstay of CC prevention in LMICs has been the single-visit approach using screen-and-treat (SVA-SAT) method, using visual inspection with acetic acid (VIA) and ablative treatment with cryotherapy to manage precancerous lesions. It is a low-cost screening approach and it minimizes loss to follow-up compared to the traditional cytology. Despite well-established effectiveness of VIA on popula- tion-level reduction in CC burden, the estimated screening uptake among women aged 30-49 in Kenya is 16%, far from the WHO's target of 70% twice-lifetime screening of women ages 35-45 by 2030. Additionally, there is extremely low fidelity of SVA-SAT; up to 70% of screen-positive women do not receive treatment. The low treat- ment rate has been attributed to programmatic and logistical challenges of implementing cryotherapy in low- resource settings (e.g., supply chain difficulties of refrigerant gas, equipment failure, and treatment duration >10 min). Thermal ablation (TA), was recommended by the WHO in 2019 and is an effective and safe alternative to cryotherapy. The portable device can be charged with electricity, batteries or solar panels, which is ideal for rural settings. However, wide dissemination and adoption have been challenged by undefined drivers of successful implementation. Our objective is to develop and evaluate a locally contextualized dissemination and implemen- tation (D&I) strategy for SVA-SAT with TA (SVA-SAT+TA) to inform national scale-up. Our hypothesis is that TA will enhance the feasibility, adoption, and sustainability of CC prevention services via SVA-SAT, compared to cryotherapy. Our multidisciplinary team proposes five-year prospective, stepped-wedge, randomized trial to implement SVA-SAT+TA in ten reproductive health (RH) clinics in central Kenya. In Aim 1, we will collaborate with multi-level (clinic, county, national) stakeholders to develop a sustainable D&I strategy to introduce SVA- SAT+TA that effectively accounts for the heterogeneity of the client, provider and system inputs. In Aim 2, we will deliver and rigorously evaluate the SVA-SAT+TA intervention at scale in public RH clinics in Kenya, using the RE-AIM (Reach, Effectiveness, Adoption, Implementation, and Maintenance) framework. In Aim 3, we will compare the cost, cost-effectiveness and budget impact of SVA-SAT+TA to SVA-SAT using cryotherapy. Together, the results from this project will improve wider implementation and scale-up of an evidence-based intervention, SVA-SAT+TA, and provide the necessary evidence to guide policy and serve as a model for CC prevention in the LMICs context.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Posttraumatic stress disorder (PTSD) and hazardous drinking (HD: heavy alcohol use and negative alcohol- related consequences) commonly develop and co-occur following trauma exposure. The individual and public health burden of PTSD and co-occurring HD is substantial. Although there are evidence-based behavioral treatments for co-occurring PTSD and HD, they have limitations that result in many individuals not accessing these treatments and limited intervention reach – i.e., existing interventions tend to be complicated, burdensome, and require contact with an in-person therapist. Thus, there is a need to develop additional, novel interventions that use evidenced-based strategies and that have greater portability and reach. There is a nascent, scant literature evaluating such types of interventions for veterans, and we know of none aimed at the general public. The current application, therefore, seeks to refine and test a brief, self-directed, intervention for individuals from the general public with PTSD and co-occurring HD that can be delivered via text-messaging. The proposed intervention draws on evidence-based strategies from cognitive behavioral therapy (CBT) for treating PTSD and co-occurring HD, and it yielded promising findings in a recent pilot study conducted by the research team. This application seeks to refine the intervention further by testing whether theoretically-driven, evidence-based strategies from basic cognitive psychology (message framing) and social psychology (facilitating growth mindsets) result in better outcomes for PTSD symptoms and HD by addressing pilot participant feedback related to avoidance and motivation. We will identify the most efficacious intervention and test it in a fully-powered RCT. Thus, Study 1, which will recruit an adult community sample (N = 500), will test the efficacy of enhancing the CBT text message intervention with message framing (protect against loss vs. imagine future gains vs. no framing) and mindsets (provide growth mindset message for using skills vs. simple reminder to use skills) via a fully-crossed 3x2 factorial design. Study 2 will build on Study 1, by conducting a fully powered (N = 333) randomized controlled trial (RCT) that will test the comparative and long-term efficacy of the most efficacious CBT-based text message intervention from Study 1 versus an assessment only control in a community sample of adults with co-occurring HD and PTSD. This proposal is innovative in its approach to adding to the menu of treatment options for co-occurring PTSD and HD and with its evaluation of the utility of integrating basic social and cognitive psychology strategies with those from CBT. If successful, it will result in an efficacious, novel, theory-driven, low burden behavioral intervention co-occurring PTSD and HD that is readily scalable. Such an intervention could be implemented as a stand-alone intervention or be integrated into stepped-care approaches, including following mass trauma exposure (natural disasters, mass casualty events, pandemics) and thus, has the potential for a substantial impact on public health.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT The newly emerged SARS-CoV-2 coronavirus has demonstrated the deadly threat of pulmonary pathogens in an exposure-naïve world with no existing vaccines or therapeutics at the ready. The development of effective vaccines has provided key prophylactic products, but therapeutics remain important due to slow and incomplete world coverage, along with emergence of resistance variants. There is especially a need for polytherapy platforms that can be deployed in formats amenable to global settings, and need for platforms that can be rapidly developed against future pulmonary threats. This project aims to develop a versatile inhalable therapeutic platform against COVID-19 disease and future coronaviruses. It is designed for nebulizer and distributable inhalation devices to maximize drug activity in the lung. The polymeric prodrug platform has recently shown strong potentiating activity against highly lethal and antimicrobial-resistant bacterial lung infections. These “drugamer” therapeutics improve the activity of pulmonary drugs by targeting them to specific cell reservoirs in the lung with high and extended dosing profiles. The inhalable platform could be used by infected patients before hospitalization, to reduce administrations by patients in crowded hospitals, and contribute a key distributable therapeutic and prophylactic modality that is needed to protect caregivers and disadvantaged populations. The proposal is structured around 4 specific aims: (1) Develop remdesivir and baracitinib as first drugamer candidates that exploit the lung macrophage as a reservoir to achieve extended dosing, as well as targeted designs against lung epithelium viral reservoirs. Remdesivir and baracitinib prodrug monomers will be developed with corresponding drugamer designs with mannose and peptide targeting ligands for the alveolar macrophage and epithelial compartments, respectively; (2) Characterize and optimize the drugamer candidates by criteria of how they load drugs into the lung macrophage and epithelial cells with extended dosing times. This will lead to better understanding of how to optimize targeting strategies in the lung for future antiviral development. The mechanisms will be studied by using quantitative LC-MS pharmacokinetics characterization and safety characterization using lung inflammatory response assessments; (3) Assess and optimize drugamer activity against SARS-CoV-2 using the hACE2 mouse model. Viral load and survival studies will be used to characterize and develop optimized drugamer and drugamer combinations that could in the future be carried forward into preclinical development. Compared to current formulation approaches, the drugamers exhibit higher drug loading, the ability to co-formulate widely varying drugs for polytherapy, and individually tailorable drug PK profiles that minimize burst release. The modularity of the platform, together with scaled and rapid manufacturing response attributes, will allow diverse incorporation of other drugs as combinations. These favorable platform attributes motivate this project to develop a new repertoire of current and future coronavirus therapeutic products.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Data from our mouse model of Botulinum Toxin A (BTxA) induced muscle paralysis has revealed that neuromuscular function, outside the axis of mechanical loading deficits, is a critical modulator of bone homeostasis. Consistent with this thesis, we have observed that transient muscle paralysis triggers acute inflammatory signaling within bone marrow that precedes the onset of focal RANKL-mediated osteoclastogenesis, which is responsible for the profound cortical and trabecular bone resorption observed in the model. However, the intercellular signaling that initiates acute bone marrow inflammation and subsequent bone resorption has not been elucidated and therefore presents a barrier to identifying translational strategies that would decouple neuromuscular dysfunction from bone loss. One potential initiator of this rapid response is neurogenic inflammation, which is triggered by neuropeptide release from sensory nerves and is amplified by mast cell mediated histamine release. We therefore pursued a series of preliminary studies to assess activation of this pathway following muscle paralysis and found that: 1) Substance P, a classic initiator of neurogenic inflammation, was upregulated in tibia bone marrow within 1 d of calf paralysis, 2) genes associated with connective tissue mast cell activation were acutely elevated following muscle paralysis, and 3) muscle paralysis induced bone resorption was significantly diminished in mast cell deficient KitW-sh/W-sh mice. We therefore hypothesize that: Bone resorption following muscle paralysis is initiated by neuropeptide signaling and is amplified by mast cell dependent histamine release. We will pursue this thesis via four complementary Specific Aims (SA), each with a corresponding sub-hypothesis. First, we anticipate that neuropeptides within bone marrow will be elevated by BTxA induced muscle paralysis prior to evidence of mast cell activation or bone resorption (SA#1). SA#2 will then demonstrate that successful antagonism of these neuropeptides will be required to inhibit mast cell activation and bone resorption induced by muscle paralysis. In SA#3, we will leverage a cKit independent, connective tissue mast cell deficient mouse to demonstrate that mast cell mediated histamine signaling is responsible for the profound osteoclastogenesis induced by muscle paralysis. SA#4 will then provide proof of concept that treatment with histamine receptor antagonists will significantly attenuate bone resorption caused by muscle paralysis. Each aspect of the proposed signaling pathway (neurogenic inflammation, neuropeptide signaling, mast cell activation, paralysis induced bone resorption) has been explored in other contexts but has not been integrated into a cellular signaling cascade that integrates muscle, nerve, and bone physiology. Importantly, if our thesis is supported, the broad clinical experience with histamine antagonists will enable repurposing of approved drugs toward the goal of ameliorating acute bone resorption precipitated by paralysis or other neuromuscular impairments.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Monoclonal antibody (mAbs) therapies have proven to be effective treatments for a multitude of clinical applications. Several of these treatments require repeated administration through intravenous infusions leading to high costs and poor patient adherence. A new field has emerged that seeks to generate mAb-secreting plasma cell therapeutics that could be administered once to provide lifelong therapeutic mAb levels as a therapeutic alternative. Plasma cells have high secretory capacities, can last a lifetime and are thought to be relatively quiescent. Advances in gene editing and ex vivo B cell differentiation have enabled the ability to generate plasma cells that secrete an exogenous mAb. However, current gene editing strategies do not fully account for endogenous antibody expression or control for what isotype the exogenous mAb is expressed as. Additionally, the field of gene-edited human plasma cell lacks both large and small animal models to test the engraftment and functionality of these mAb-secreting plasma cells. Aim 1 of this proposal will use a new gene engineering strategy to generate a human plasma cell therapeutic that secretes an engineered IgG mAb against Epstein Barr Virus (EBV) as a proof of concept for future mAb- secreting cell therapies. I will characterize the ability of gene-edited primary plasma cells to produce large quantities of defined mAbs and compare to previously published methods. Human plasma cells are difficult to engraft in small animal models due to the lack of extrinsic survival factors from human stromal and myeloid cells. Aim 2 of this proposal will use a humanized (NSG-huCD34) mouse model to better predict the potential engraftment potential of gene-edited plasma cells therapies. This humanized mouse model also serves as a model for human-tropic EBV infection. I propose to validate the functionality of anti-EBV secreting plasma cells by showing that these cells protect humanized mice against high dose intravenous EBV challenge. This would be the first time that gene-edited primary human plasma cells were shown to protect against a human-tropic disease in a humanized mouse model and would serve as a proof of concept for use of gene-edited plasma cells to provide mAbs. This project is ideal for my training as a young physician scientist due to my strong interest in translational immunology, the complementation of my previous training in chimeric antigen receptor- modified T cell research, the extensive background of Dr. David Rawlings and Dr. Richard James in B cell biology and mentorship, and the quality research and medical education at the University of Washington. The activities detailed in this proposal will provide a strong background for a future career as a physician scientist pursuing translational immunology research.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY/ABSTRACT This project aims to develop a culturally responsive technology tool for collaborative health research results dissemination with diverse American Indian and Alaska Native (AIAN) communities. New methods and technologies for respectful and culturally responsive research results dissemination are urgently needed both to help researchers engage diverse participants and to serve those communities better. For many researchers, the primary methods of disseminating research results are to submit manuscripts to peer-reviewed journals or abstracts for conference presentations which essentially limits their audience to academic scholars. Limiting dialogue between community stakeholders and researchers, many of whom may be unacquainted with community perceptions of health or community systems of sharing information, decreases the likelihood that study results will be implemented at a community level which can have a negative impact on community health outcomes. Reliable health research information supports wellness but if that information is not presented using methods that are understandable and contextualized, they have less benefit. AIAN people have experienced a history of distrustful health research practice such as being exposed to procedures without informed consent and having research results disseminated without community input or approval. Oversights like these increase the need for transparency and accountability in research at all stages from project conceptualization to results dissemination. Moreover, AIAN have been under-represented in large national health research initiatives—such as NIH’s All of Us research program—or have not been consulted in meaningful ways about how best to implement health research or disseminate actionable findings relevant to local community context. Though there has been progress with community engagement in health research, only limited research has explored collaboration specific to the results dissemination process. This study involves the co-design, co-development, and evaluation of a stakeholder collaboration tool for health research results dissemination with AIAN communities. For Aim 1, we will use value-sensitive design and user-centered design approaches to establish value informed criteria to support activities for Aim 2A. In Aim 2A, we will integrate digital storytelling techniques with participatory design methods to co-design low fidelity prototypes for collaborative results dissemination. Aim 2B will involve pilot implementation of a prototype designed with data obtained from Aim 2A which will be evaluated for feasibility and acceptability in AIAN communities. Success in this research will result in improved research communication and enhanced trust in health research which will increase the impact of health research overall by ensuring that research outcomes are effectively disseminated to AIAN communities. This will support the community in understanding and possibly incorporating research recommendations.

NIH Research Projects · FY 2025 · 2021-09

Schizophrenia is a common and devastating neurodevelopmental disorder that has defied diagnostic and therapeutic advances due to its complex pathogenesis and genetic heterogeneity. Experimental approaches that integrate genetics and biology are necessary to further our understanding of this complex disorder. Both common genetic variants of small effect and rare genetic mutations of severe effect have been implicated in schizophrenia, with the latter more likely to produce phenotypes that can be measured in vitro. This proposal describes a 5-year career development program through which I will develop the conceptual framework and gain the skills necessary to characterize rare, severe mutations in persons with schizophrenia. My project will be to evaluate, using CRISPR-interference (CRISPRi) in iPSCs, genes that share 3 criteria: 1) each gene harbors a damaging mutation in one or more patients from our studies; 2) each gene is significantly associated with schizophrenia per criteria of the SCHEMA consortium; and 3) each gene plays a role in chromatin remodeling or transcriptional regulation. In Aim 1, I will use CRISPRi to knock down expression of each of these genes in iPSC-derived neurons, then compare transcriptional signatures before and after loss of gene expression, and to other iPSC models of schizophrenia. I will also assay for changes in cellular oxidative stress, a cellular phenotype of iPSC models of schizophrenia. In Aim 2, I will generate a patient-derived iPSC line for one of the genes from Aim 1, then create an isogenic iPSC line with the reversion to wild-type of the mutation using CRISPR-mediated homology directed repair. (Lymphoblast lines of all patients from Aim 1 are available for this purpose.) I will compare patient and revertant iPSCs with the same assays as in Aim 1, and also for their ability to form mature neurons, their neurite number, cellular migration defects, transcriptional profiles, and epigenetic effects. In Aim 3, from my clinic, I will enroll additional patient-parent-parent trios; collect blood for DNA, RNA, and the establishment of cell lines; organize the collection of longitudinal clinical data; and carry out exome sequencing and identify damaging variants for my future studies. I have obtained approval of this aim from the UW IRB. This K08 award will provide me with the mentorship and conceptual and experimental training necessary for each of these approaches. My overall goal is to establish my independent laboratory studying the genes responsible for the pathophysiology of schizophrenia.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Chronic pain is a prevalent, disabling problem affecting as many as 50% of men and 75% of women Veterans. The psychosocial pain treatment with the most empirical support is cognitive behavior therapy (CBT). However, not everyone benefits from CBT. Prior research has shown that two complementary and integrative health (CIH) interventions, Mindfulness-Based Cognitive Therapy (MBCT) and Hypnotic Cognitive Therapy (HYP-CT), are beneficial and offer alternatives to CBT and have similar efficacy. As with CBT, individual response to these treatments also varies considerably. Our preliminary data suggests the variability in treatment outcome may be due to important baseline patient differences. This suggests the possibility that patients can be effectively “matched” to the treatments they are most likely to benefit from, based on information that can be obtained before treatment starts. Research is needed to develop patient-treatment matching algorithms that could match a given individual to the evidence-based treatment most likely to be beneficial given their unique individual profile. The overall aim of the proposed study is to address this need by identifying patient factors that determine who benefits most from two CIH treatments and the current gold standard non-pharmacological treatment (CBT), relative to usual care. This aim will be addressed using a clinical trial in which participants will be randomized to one of the four treatment conditions. We will test hypotheses – developed from both theory and prior pilot research – that pre-treatment brain activity patterns, hypnotizability, catastrophizing, and mindfulness moderate pre- to post-treatment improvements in pain intensity associated with group-delivered CBT, MBCT, and HYP-CT, relative to usual care, in Veterans with chronic pain. The findings will provide the foundation for building an algorithm to support personalized care, whereby Veterans can be offered the treatment that will be most beneficial for them. This study will also help develop knowledge about potentially powerful alternatives to pharmacological analgesic approaches. Finally, the study findings will be useful to health care providers and their patients to facilitate shared decision-making.

NIH Research Projects · FY 2025 · 2021-09

Project Abstract The specific goal of this proposal is to develop and optimize methods for quantitative pre-clinical PET imaging for immune checkpoint inhibitor (ICI) therapies in non-small cell lung cancer (NSCLC). We will leverage an existing co-clinical trial using our genetically-engineered mouse model (GEMM) of lung adenocarcinoma to develop, test and implement the methods and populate a web-accessible research resource. This web- accessible research resource will in turn leverage recent developments in quantitative cancer imaging informatics using the industry-standard DICOM format. In response to PAR-18-841 there are four components to our proposal: (1) appropriate models, (2) a co-clinical trial including a therapeutic goal, (3) quantitative preclinical and clinical PET imaging, and (4) an innovative quantitative cancer imaging informatics platform. Our longer term goal is to improve outcomes for NSCLC patients, as lung cancer is still the leading cause of cancer deaths worldwide. Although ICI therapy has been a tremendous clinical benefit for some NSCLC patients, only ~20% of NSCLC patients respond to anti-PD1/PDL1 therapy. Using PET co-clinical imaging to improve ICI therapies for NSCLC is challenged by a lack of suitable informatics methods to capture and track necessary meta-information, appropriate response criteria for preclinical imaging, which in turn is a consequence in part of the lack of quantitative preclinical PET imaging methods that are linked to quantitative clinical PET imaging methods. We will address these challenges with three Specific Aims: 1, Develop and optimize quantitative preclinical quantitative imaging methods and protocols. These methods will involve novel long-lived phantoms that can cross-calibrate multiple preclinical and clinical PET scanners. This will be tested with partner members of the Co-Clinical Imaging Research Resources Program. 2. Implement the optimized methods in our co-clinical trial of ICI treatment of NSCLC with a GEMM lung adenocarcinoma model and a GEMM model lung squamous cell carcinoma. From this we will evaluate how the information gleaned from the pre-clinical and clinical studies can be used to inform future pre-clinical studies in terms of optimized mouse imaging protocols and response criteria. 3. Share data and resources on co-clinical trials using quantitative PET imaging using a web-accessible open science approach (open source + open data) by extending the DICOM standard for pre-clinical small animal imaging with DICOM-compliant structures that provide necessary quantitative meta-data. These methods and resources developed during this project will be distributed to accelerate the development of needed effective cancer therapies by improving the utility of early-phase oncology trials using co-clinical studies with PET imaging. In addition, we will determine, and potentially improve, the utility of PET imaging as a biomarker for early assessment of response in co-clinical immunotherapy studies by using an appropriate mouse model.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Benzalkonium chlorides (BACs) are widely used antimicrobials in disinfecting products, medical products, consumer products, and food processing industries, suggesting humans may be exposed chronically and sys- temically to BACs through a variety of routes. Our preliminary study found that close to 50 of 100 random hu- man plasma samples contain detectable levels of BACs, suggesting BACs are indeed absorbed. The ongoing COVID-19 pandemic has led to greatly increased use of BAC-containing disinfectants, which is concerning given accumulating evidence in respiratory, developmental, reproductive, and neurological toxicities inflicted by BACs and BAC-induced disruption of cholesterol and lipid homeostasis in rodents. However, there is a lack of knowledge on the metabolism, transport, and biological consequences of BACs in humans. Our goal is to characterize the pathways of metabolism and transport of BACs and their impact on nephrotoxicity of BACs. The potential for nephrotoxicity is supported by previous studies in rats showing that BACs accumulate to the highest level in the kidney after oral intake and our preliminary studies showing that BACs exert potent cytotox- icity in a 3D “kidney-on-a-chip” microphysiological system (MPS). Recently, we reported that BACs are metab- olized by human cytochrome P450 (CYP) isoforms CYP2D6 and CYP4s in vitro. Furthermore, we found that BACs are actively transported by human organic cation transporters (hOCTs). Because CYP2D6, CYP4s, and hOCTs are highly polymorphic with greatly varying protein activities, we hypothesize that toxicities of BACs in kidney are dependent on the activities of BAC-metabolizing and transporting proteins in both liver and kidney. In Aim 1, we will characterize pathways of metabolism and transport of BACs in vitro, including secondary me- tabolism by β-oxidation and glucuronidation and transport by hOCTs, human multidrug and toxin extrusion pro- teins, and P-glycoprotein. In Aim 2, we will evaluate nephrotoxicity induced by BACs in human proximal tubule epithelial cells in 3D integrated liver-kidney “organs-on-chips” MPS. An integrated sterolomics, lipidomics, and transcriptomics approach will be used to systemically assess the toxicity and biological activities of BACs. In Aim 3, we will assess BAC exposure levels and their correlation with lipid and kidney injury biomarkers in hu- mans, as well as the impact of genetic variations on BAC metabolism and disposition. The significance of this project lies in that it will, for the first time, address the knowledge gap in metabolism, transport, and toxicity of BACs in humans. Elucidation of the contribution of reduced activities in CYPs and/or transporters to BAC tox- icity would enable us to identify high-risk human population with genetic variations in these proteins. The gained knowledge could also inform federal agencies on setting more appropriate exposure limitations. The innovation of this project lies in a) a novel example of gene-environment interaction through xenobiotic- processing proteins, b) the use of an integrated liver-kidney MPS to assess the toxicological consequences of xenobiotics, and c) integrated omics for rigorous systems toxicology studies.

NIH Research Projects · FY 2024 · 2021-09

Project Summary This proposal delineates a 4-year research career development program focused on brain mechanisms by which sex steroids protect from atherosclerosis. The candidate is currently a Research Assistant Professor in the Department of Medicine, Division of Metabolism, Endocrinology and Nutrition at University of Washington (UW). The proposed experiments and didactic work will provide the candidate with a unique set of multi- disciplinary skills for him to become an independent investigator in the cardiovascular field. The candidate has completed a PhD degree in Pharmacology at the University of Chile and two postdoctoral fellowships at OHSU and UW. The applicant’s combination of expertise in sex steroids biology and CNS regulation of metabolism uniquely qualify him to conduct these studies. In this proposal, he will address key questions in the field of cardiovascular disease, the answers to which will increase our understanding of the relationship between androgen deficiency and elevated cardiometabolic risk. Dr. Dorfman and his mentors (Drs. Karin E. Bornfeldt and Joshua P. Thaler) have created a specific training plan that takes advantage of the intellectual resources at the UW to guide him through the proposed educational activities and research. The goal of the current work is to work in mouse models to identify CNS mechanisms by which hypogonadism increases the incidence of cardiovascular events. Specifically, he will investigate the role of glial cells (microglia and astrocytes) in cardiometabolic changes that accelerate the progression of atherosclerosis. The proposal focuses on human evidence that low levels of circulating androgens promotes cardiometabolic dysregulation, and that hypothalamic gliosis is associated with lower endogenous testosterone and plasma LDL in men. Furthermore, the preliminary data identify a dramatic synergistic increase of hypothalamic gliosis when hypogonadism induced by castration and Western-type diet are combined; in this animal model, early markers of cardiometabolic risk are also evident including increased liver inflammation and triglyceride content, a shift toward very low density lipoprotein species, and circulating leukocytosis. These findings support studies proposed in Specific Aim 1 to use animal models with ablation of microglial or astrocyte inflammation to determine whether hypothalamic gliosis is required for Western-type diet and androgen deficiency to synergistically predispose to atherosclerosis. Additionally, the candidate has developed and validated a mouse model in which microglia and astrocytes can be pharmacologically activated through the use of Designer Receptors Exclusively Activated by Designer Drugs (DREADD) technology. These models of glial activation provide a tool to determine whether gliosis is sufficient to increase the risk of CVD as proposed in Aim 2.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY / ABSTRACT Tuberculosis (TB) is the leading infectious killer worldwide. Upon exposure to Mycobacterium tuberculosis (Mtb), most people develop asymptomatic latent TB infection (LTBI phenotype). However, Dr. Anterasian's collaborators identified about 7% of individuals who, despite household exposure to pulmonary TB, do not convert their tuberculin skin test (TST) or interferon-γ release assay (IGRA), and thus can be classified as clinically resistant to traditionally defined LTBI (“RSTR” phenotype). Uncovering mechanisms of natural resistance to Mtb infection may provide unique insights that can inform the development of host-directed therapeutics (HDTs). Dr. Anterasian has analyzed the first global proteomic dataset of Mtb-infected primary human macrophages and discovered 46 differentially abundant proteins (DAPs) that define the RSTR vs LTBI macrophage proteomic response to Mtb. By integrating her data with those of her collaborators, Dr. Anterasian has identified DAPs that may undergo Mtb-induced post-translational modifications (PTMs) and/or interact with Mtb bacterial proteins. She has also used bioinformatic network analyses as well as genetic and transcriptomic data from the same patient cohort to ultimately curate a list of 19 proteins for further mechanistic studies. In particular, the Rab family of GTPases interact with Mtb bacterial proteins, undergo Mtb-induced ubiquitination, and are key regulators of membrane trafficking and autophagy, which are pathways Mtb exploits during infection. The objective of this proposal is to define macrophage pathways and proteins that characterize the protective RSTR response. Dr. Anterasian hypothesizes that RSTR individuals promote macrophage Mtb clearance through DAP-mediated pathways modulated by DAP-Mtb protein interactions and differential DAP ubiquitination. In Aim 1, Dr. Anterasian will investigate Rab-dependent mechanisms of protection against Mtb in macrophages and how Mtb proteins subvert Rab function. In Aim 2, Dr. Anterasian will identify candidate proteins associated with control of Mtb infection, their PTMs, and ubiquitin-dependent mechanisms of resistance. By combining proteomic, bioinformatic, and cellular immunology approaches, Dr. Anterasian is well poised to identify key pathways in human macrophage resistance to Mtb that can be targeted with HDTs. Dr. Anterasian is a Pediatric Infectious Disease Fellow in the Division of Infectious Diseases at Seattle Children's Hospital and the University of Washington. She additionally proposes a comprehensive career development program that includes: 1) mentored training in proteomics and cellular immunology; 2) formal didactics in large data set analyses (i.e. proteomics, statistical genetics, systems biology); 3) mentorship in the design of proteomic studies, human subjects research, and scientific writing; 4) structured opportunities to present her work to local and international scientific audiences; and 5) Scientific Advisory Committee meetings that ensure scientific and career development progress. By the conclusion of this award, Dr. Anterasian will transition to an independently-funded expert in the Mtb host response who will direct her own laboratory.

NIH Research Projects · FY 2024 · 2021-09

Project Summary / Abstract The blood protein von Willebrand factor (VWF) is a large multimeric protein that, when activated, binds to blood platelets tethering them to the site of vascular injury initiating blood coagulation. This process is critical for normal haemostasis, but especially under inflammatory conditions it is thought to be a major player in patho- logical thrombus formation. For this reason, VWF has been the target for the development of anti-thrombotic therapeutics. However, a particular challenge is how to prevent pathological thrombus formation while still allow- ing normal physiological blood coagulation. In fact, currently available anti-thrombotic therapeutics are known to cause intracranial bleeding as side effect. The work presented here proposes that by identifying the confor- mational changes that VWF undergoes in inflammation it will be possible to design molecules that target VWF selectively only in an inflammatory pro-thrombotic environment. Experiments performed in vitro have indicated that oxidizing agents released during inflammation increase the platelet-binding activity of VWF, and this has been linked to the oxidation of methionine residues within VWF. This study aims to characterize what inhibitory mechanisms are removed in VWF through methionine oxidation and identifies sites that can be targeted optimally under oxidizing conditions. We hypothesize that oxidation activates the A1 domain (the domain in VWF that contains the binding site to platelets) by removing the masking function of its neighboring domains. Through a combination of a dynamic flow assay, equilibrium unfolding experiments and a binding assay, we will identify which auto-inhibitory mechanisms of VWF are turned off by oxidation and which methionine residues are key for activation under oxidizing conditions. This knowledge will then be used in computational docking studies to create a model of the complex between the A1 domain and its neighboring domains and study the structural effects of methionine oxidation at atomic level of detail. An assay using samples from patients with inflammatory conditions will be used to test the link between inflammatory conditions and higher activity of VWF in vivo. The structural insights gained here will be invaluable to structure-based drug design in order to develop therapeutics that target VWF only when it is in its oxidized state in order to prevent thrombosis while allowing haemostasis.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Though trauma is often considered an isolated disease process, advancements in acute care have shifted the burden of injury from the acute physiologic insult to the chronic post-injury sequalae. Over 95% of hospitalized trauma patients now survive to discharge. However, many go on to suffer long-term consequences of injury in the form of worsened functional status, mental health, and health-related quality of life. These trauma-initiated chronic conditions make it difficult for many working-age adults to return to work and economic productivity. The combination of poor clinical outcomes and work loss may be further compounded by unexpected medical debt as many working-age adults carry high-deductible insurance plans. Unfortunately, there is a paucity of information regarding long-term clinical and economic outcomes after major injury in the US. As a result, the policy interventions needed to optimize long-term recovery after major injury remain unknown. The purpose of this proposal is to close these key knowledge gaps and map a policy response moving forward. The specific research aims of this proposal are to evaluate (i) long-term clinical outcomes and (ii) long-term economic outcomes among working-age survivors of traumatic injury; and to evaluate (iii) the impact of alternative insurance design strategies on long-term outcomes after traumatic injury. The proposed study leverages a unique data opportunity at our institution and will be the first of its kind to combine a state-wide trauma registry (containing both inpatient data and long-term patient-reported outcomes) with 24 months of pre-injury and 24 months of post-injury consumer credit reports. We will use the evidence from these novel analyses to simulate changes in insurance benefit design in order to optimize return to work and mitigate financial strain after injury. This project will also facilitate the career development of the candidate. The highly experienced and multidisciplinary mentorship team, the proposed career development and research plan, and the unparalleled research environment are ideally suited to address the career goals and educational needs of the candidate PI, John W. Scott, MD, MPH. Although he has substantial prior experience in surgical health services research, he lacks experience with (i) longitudinal data analysis, (ii) evaluating personal financial data, and (iii) policy simulation methods. In addition to the innovative research plan, this proposal also includes additional training which will be essential both for the successful completion of this research and for Dr. Scott’s career development, including graduate level courses and experiential learning opportunities. This career development award will lay the groundwork for Dr. Scott to mature as an independent investigator and national leader in improving long-term clinical and economic outcomes after acute illness and injury through policy evaluation and reform.

NIH Research Projects · FY 2024 · 2021-09

Alzheimer’s disease and other dementias represent an increasing burden on society, with an estimated 5.8 million Americans suffering from Alzheimer’s disease. A major barrier to progress in Alzheimer’s disease research is a lack of suitable animal models that (1) show pathological hallmarks and clinical features similar to those seen in patients with Alzheimer’s disease, and (2) have similar genetic and environmental heterogeneity to people. The privately-owned companion dog uniquely captures both of these features. The Dog Aging Project (DAP) is a consortium of investigators with the shared goal of understanding the biological aging process, including age-related cognitive changes and dementia, in companion dogs through large-scale longitudinal study and clinical evaluation of putative healthspan-promoting interventions. More than 50,000 companion dogs will ultimately be enrolled in the DAP Pack, for which detailed owner survey information is collected annually, including the gold standard cognitive assessment questionnaire for diagnosis of canine cognitive dysfunction (CCD). A high-resolution “Precision Group” of 1,000 dogs are being studied in much greater depth, including full genome sequencing, veterinarian-reviewed electronic medical records (VEMRs), and annual assessments including physical exam, clinical chemistry, blood epigenome, serum metabolome, and fecal microbiome. We propose to synergistically leverage the infrastructure of the DAP to create an unparalleled and one-of-a-kind resource for studying Alzheimer’s-like disease in the companion dog. To accomplish this goal, we will (1) recruit 200 dogs with CCD into a “CCD Precision Group” which will be studied at high resolution in parallel with the cognitively normal DAP Precision Group, including assessments of serum abundance of AD markers such as Ab42, Tau, and hyperphosphorylated Tau, (2) quantitatively assess proteomic and neuropathological hallmarks of Alzheimer’s-like disease in brains from 100 companion dogs who reach the end of their natural lives, and (3) create a large canine data and biospecimen repository to support future studies of Alzheimer’s-like disease in companion dogs. We anticipate that successful completion of this project will not only create a rich dataset on AD-like disease in companion dogs, but will also spur numerous follow-on studies by other investigators. It is our hope and expectation that these resources will have a major impact on Alzheimer’s disease research far into the future.