Oregon Health & Science University

NIH Research Projects · FY 2025 · 2024-08

Project Summary The purpose of this application is secure funding for a 5-year period to support travel awards to enable US-based registrants from diverse backgrounds that are selected in an equitable manner to attend the Oregon Health & Science University (OHSU) annual Developmental Origins of Health and Disease (DOHaD) Summer Course, which is held the first week of August each year at OHSU. The target demographic is graduate students, post-doctoral fellows, and early career investigators who have a primary interest in the fundamentals and the latest advances in the DOHaD field, but who may not have the resources to attend the week-long course. Attendance will further provide the opportunity for them to interact with leaders in the DOHaD field and each other, thus fostering connections among the next generation of scientists and leaders. The long-term objective is to continue to expand DOHaD and placental-related research nationally, develop interactive collaborative networks in this field, and support the development of early career scientists. The meeting will feature lectures from OHSU faculty members who work in the DOHaD area, that cover the fundamental principles and cutting edge work in thematic areas (e.g. cardiovascular, neurodevelopment) relevant to DOHaD. On day one, a “connections corner-like” event will allow attendees to interact with each other and faculty. Attendees will be assigned a mentor for the meeting and will take part in an experimental design challenge where they will design specific aims and experiments to address a research question for presentation at the end of the conference. In addition, all registrants are required to submit an abstract describing their own work for poster presentation on day two of the meeting. The R13 funded travel awards will be selected by a competitive review of these abstracts by the organizing committee and ranked according to clarity and content. During award selection, the committee will give special regard to representation of minorities and claims for financial support. In addition, this grant will sponsor one Keynote lecture by an accomplished scientist outside of our institution to provide attendees with state-of-the-art perspective on a leading issue in DOHaD. This will be an individual who is not necessarily a DOHaD scientist, but is clearly a leader in an area that has relevance to DOHaD (e.g. angiogenesis, apoptosis, cancer biology). This Keynote Speaker will serve as a focal point for a plenary session and workshop in that area. This individual will be designated the NIH-Lecturer at each meeting.

NIH Research Projects · FY 2026 · 2024-08

Cryo-electron microscopy (cryo-EM) has transformed the biomedical field by enabling researchers to examine molecular biology components with unparalleled detail. In 2018, the National Institutes of Health (NIH) established a national network of cryo-EM centers, including the Pacific Northwest Center for Cryo-EM (PNCC), to meet the growing demand for access and training in this groundbreaking technology. Over the past five years, PNCC, along with partner centers in New York (NCCAT) and Stanford (S2C2), have become an indispensable resource for the structural biology community. The field of cryo-EM continues to advance at a tremendous pace, driven by technological breakthroughs in instrument design, computational algorithms, and specimen preparation methods. It is expected that developments in machine learning and artificial intelligence will also play a significant role in enhancing cryo-EM workflows and automating various aspects of the process. These advancements in the field of cryo-EM are enabling researchers to study increasingly complex biological systems, capture low-abundance targets, analyze time-resolved reaction pathways, and obtain precise details of biomolecular interactions. The continued growth in cryo-EM promises to facilitate discoveries in basic sciences, biomedical research, and the pharmaceutical industry for years to come. With the rapid technological progress comes an increasing demand for access to high-end cryo-EM instrumentation and training opportunities. It is therefore essential to ensure access to resources and training to investigators entering the field. Additionally, support is needed for established researchers in applying cutting-edge technologies to challenging high-value structural biology targets. The PNCC is committed to meeting the evolving needs of the cryo-EM community through effective and innovative approaches in service, training, and outreach. PNCC benefits from the partnership between Oregon Health & Science University (OHSU) and Pacific Northwest National Laboratories (PNNL), both of which provide unique expertise, resources, and infrastructure. The highly experienced operational team at PNCC is adept at managing center resources, adapting to evolving developments, and providing high-quality training in cryo-EM methods to both new and experienced cryo-EM practitioners. Working with its partners, PNCC will position the center at the forefront of the field, develop and implement new and efficient workflows and training programs, and engage with the community to ensure that the benefits of all advances are widely disseminated. To expand the reach of cryo-EM, PNCC will continue to pursue innovative, sustainable approaches to overcome barriers of access and utilization. The future holds exciting opportunities and challenges for PNCC as it continues to pursue its missions and contribute to the advancement of cryo-EM.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Influenza causes an estimated 1 billion cases yearly, with severe cases leading to fatal bronchopneumonia, particularly in vulnerable populations. Influenza virions express two major surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA), the latter of which is of interest due to its role in viral egress and cleavage from mucus. Currently, the only FDA-approved antivirals used to treat influenza are neuraminidase inhibitors, which are ineffective against some strains. Monoclonal antibodies (mAbs) directed at NA provide an alternative therapeutic candidate. Delivering NA mAbs to the respiratory mucosal surface, where the virus replicates, is a novel delivery method that could improve drug efficacy and patient compliance. This five-year research career development award will provide training and development of the skills necessary for the candidate to establish an independent research laboratory focused on understanding how human antibodies against influenza can be harnessed to treat and prevent disease and transmission in animal models. Currently, the candidate is a veterinarian and a graduate trainee at the Vanderbilt Vaccine Center who will transition to an assistant professor position on the physician-scientist track in July 2024. Her training to date has focused on the isolation and characterization of human monoclonal antibodies (mAbs) from subjects with prior influenza history and mouse models of therapy. She will supplement this experience with further training in developing in vivo ferret and ex vivo human tissue models of influenza infection with which to study the natural pathogenesis of this virus and how human mAbs bind influenza to reduce pathogenesis. The short- term goals of the proposed studies are to test the central hypothesis that neuraminidase-directed antibodies play a significant role in the protective immune response and can be harnessed with antibody therapeutics. This training will be supported by mentoring from national experts in the study of human antibodies (James Crowe, Jr., Ivelin Georgiev), human airway epithelial cultures (Timothy Blackwell), influenza vaccinology (Spyros Kalams), and small animal models of virus infection and pathology (Katherine Gibson-Corley). The applicant aims to 1) elucidate the mechanisms of influenza type B (IBV) inhibition by mAbs specific to the neuraminidase (NA) glycoprotein to test the hypotheses that NA antibodies exert distinct mechanisms of protection, 2) use a human airway model of IBV infection to determine how NA mAbs in the presence of mucus behave, 3) and test these mAbs in a ferret model of intranasal delivery to test the hypothesis that NA mAbs delivered topically can reduce viral load and shedding. Overall, these studies will help define the role of IBV NA antibodies and the mechanism of delivering mAbs to the respiratory mucosal surface. Industry groups are developing protective mAbs made by the candidate as human therapeutic agents. Thus, these studies directly translate the importance of human health and contribute to a more fundamental understanding of the disease.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY / ABSTRACT Current antiretroviral therapy (ART) regimens have rendered HIV a manageable chronic condition rather than a fatal disease, with people who initiate ART soon after infection achieving nearly normal life expectancy. As a consequence of increased survival, more women living with HIV (WLWH) will now undergo menopause and be subject to a disease burden that reflects age along with adverse skeletal consequences of postmenopausal estrogen deficiency. In addition to known bone loss and increased fracture risk in postmenopausal women, ART also causes negative impacts on bone. Therefore ART-suppressed WLWH may have a compounded detrimental effect on bone due to long-term ART treatment combined with menopause. We hypothesize that treatment with estrogen replacement will attenuate the increased rate of bone loss during menopause and improve bone health even with congruent administration of ART. We will address this hypothesis using a previously NIH-funded project that employs a novel non-human primate model that replicates the sequence of acute infection, ART initiation, chronic suppressed infection, and subsequent menopause now characteristic of a growing proportion of aging WLWH. We intend to measure the impact of ART-suppression and menopause, with and without hormone replacement therapy, on bone architecture and quality. Furthermore, we will determine whether bone marrow, a known HIV reservoir, is deficient in necessary bone cell precursors. In conclusion, these studies will examine bone health in an aging population of WLWH and potentially inform future care options utilizing hormone replacement therapy to protect these women from compounded risk of bone fractures and osteoporosis.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The progression from a pluripotent state into the myriad cell types that arise during mammalian development involves a host of epigenetic properties that dictate gene expression patterns and enforce the commitment of a cell to a specific lineage. The advent of single-cell genomics has enabled the study of cell lineage dynamics at unprecedented resolution; however, such studies have been largely limited to RNA transcription. The dynamic modulation of the epigenome that governs these processes involves multiple layers of regulation, including a major role by DNA methylation – a canonically repressive property that can act as a gatekeeper of genomic permissiveness. Aberrant DNA methylation is also associated with numerous developmental disorders and diseases; however, it is also one of the least studied properties, particularly at the single-cell level, primarily due to technological limitations. We have recently addressed this shortcoming with the development of a high- throughput single-cell DNA methylation assay capable of producing 100’s of thousands of cell profiles in a single experiment, paving the way for studies of this property with a power comparable to that of transcription. Here we propose the production of at atlas of DNA methylation during mammalian embryonic development in the mouse at 24-hour increments from embryonic day 8.5 to 17.5 in order to produce a granular map of DNA methylation dynamics during key stages of lineage priming and commitment. We will further assess the intestinal epithelium using novel multiomic technologies in order to gain mechanistic insight into how layers of epigenetic control drive and enforce one another. These data will enable the assessment of standing hypotheses regarding the role of DNA methylation in genomic permissiveness and cell fate decisions. Finally, the datasets, which will represent a powerful resource for developmental biology and epigenomic studies, will be made openly available through interactive portals to enable broad access by the greater research community.

NIH Research Projects · FY 2025 · 2024-08

SUMMARY The overall goals of this U01 application are two-fold. First, to investigate two potential KSHV targets, one structural and one non-structural, to determine if these two targets are capable of protecting vaccinated animals from infection. And, the second, to compare two vaccine strategies to stimulate immune responses to the KSHV antigens. The first approach utilizes mRNA-lipid nanoparticles (mRNA-LNPs) vaccines, the widely used vaccine strategy that has proven capable of inducing protective immunity against SARS-CoV2. The second approach employs a novel vaccine strategy that targets CD180, a toll-like receptor (TLR) expressed on antigen presenting cells (APCs) that can stimulate robust immune responses to conjugated antigens, even in the context of immune suppression. To accomplish these goals, we will utilize the rhesus macaque rhadinovirus (RRV)/rhesus macaque (RM) model of KSHV-like infection to evaluate the KSHV targets and compare the immune responses induced by the mRNA-LNPs and targeted CD180 vaccine platform. The innovation and strengths of this application are several and include the collaboration formed amongst the principal investigator, and co-investigators from the biotechnology sector (Abacus Bioscience), whose company's focus is on a novel targeted CD180 vaccine platform (anti-CD180) capable of inducing robust immune responses to conjugated antigens in rodents and nonhuman primates (NHPs), and academic co-investigator, whose expertise is on the characterization of primate antibody-mediated function. Importantly, data presented in the preliminary studies section supports this application as 1) CD180 is expressed on multiple B cell lineages, dendritic cells and monocytes, similar to that shown in humans. 2) fusion of an antigen to agonist anti-CD180 antibody (aCD180) provides the simultaneous antigen delivery and activation of the APC, resulting in a potent antigen-specific IgG production and expansion of activated antigen-specific T cells in immunocompetent and immune suppressed animals. 3) we have successfully expressed two recombinant aCD180-KSHV antigen complexes (aCD180-K8.1 ectodomain and aCD180-KSHV vIL6) to evaluate in vitro and in vivo. 4) we have established an oral challenge model for RRV infection, that results in virus infection. 5) we have created the necessary chimeric KSHV/RRV recombinants pseudotyped with KSHV glycoprotein K8.1 or a recombinant RRV encoding KSHV vIL-6 in place of RRV vIL-6. These chimeric RRV recombinants can infect and establish latent infections in RM, identical to wild-type RRV. Thus, the overall goals are to compare state-of-the-art mRNA-LNPs to the novel anti-CD180 platform to stimulate an anti-K8.1 and anti-vIL6 immune responses in immune competent and SIV-infected RM prior to oral RRV challenge/infection. The overall hypothesis is immunization targeting KSHV proteins will develop cell and humoral immune responses against viral antigens. This direct head-to-head comparison serves as a necessary prerequisite to evaluating either vaccine platform in RM prior to clinical evaluation in humans.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY / ABSTRACT Cervical cancer is the fourth most common cancer affecting people with a cervix worldwide, after breast, colorectal, and lung cancers, with 569,847 new cases every year. In the United States, cervical cancer incidence and mortality are 7.7 and 2.3 per 100,000 women, respectively, with substantial disparities in both cervical cancer incidence and mortality across racial and ethnic groups. Latinas, in particular, are disproportionately burdened with cervical cancer compared to non-Hispanic Whites (NHW), with incidence and mortality rates 50% higher than NHWs. Among women diagnosed with invasive cervical cancer, Latinas are more likely than NHWs to be diagnosed at an advanced stage, when there are fewer treatment options. Despite the average age of diagnosis of all cervical cancers occurring around age 50, the majority of screening interventions focus on younger and/or reproductive aged people with a cervix. Thus, there is growing urgency for improving cervical cancer prevention among middle to older age women nationally and internationally. In the U.S., middle to older age immigrant Latinas are disproportionately likely to be served at settings like Community Health Centers (CHCs). In 2017, 56% of poor and foreign-born Latina/os lived within two miles of a CHC. CHCs provide critical infrastructure and offer healthcare to historically underserved populations (e.g., those without insurance). Evidence also suggests that there are fewer racial/ethnic healthcare disparities among patients served by CHCs, underscoring the importance of studying and understanding the care delivered in these settings. Therefore, the goals of the proposed research, as part of a career development award, are to (1) describe cervical cancer screening rates in a 23-state network of CHCs from 2012 to 2023 among middle to older age Mexican immigrants; (2) examine individual, organizational, and structural level facilitators and barriers to cervical cancer screening among middle to older age Mexican immigrants in three CHCs located in the Pacific northwest; and (3) co-develop a multi-level intervention to improve cervical cancer screening among middle to older age Mexican immigrants in CHCs. This career development award will establish Dr. Vasquez Guzman as a multi-disciplinary investigator focused on implementing and disseminating novel primary care multi-level interventions that target complex, underserved patient populations to reduce disparities in ongoing and regular timely and high-quality cervical cancer prevention and screening services in safety-net settings. The K01 award provides mentored training for Dr. Vasquez Guzman to develop expertise in 4 areas: (1) team science and the integration of social science and clinical research approaches; (2) skills and knowledge in both intervention and implementation science (3) cervical cancer screening and cancer communication skills; and (4) expand community engaged expertise. To achieve these goals, Dr. Vasquez Guzman has assembled a well-funded and established mentoring team with expertise in health services research, primary care, intervention design and implementation science, team science, and CBPR, all of whom have success in mentoring faculty to research independence.

NIH Research Projects · FY 2026 · 2024-08

PROJECT SUMMARY The United States faces an unprecedented mental health crisis among children and adolescents. In 2021, 29% of high school students reported experiencing poor mental health. Almost 60% of female students experienced persistent feelings of sadness during the past year, and nearly 25% made a suicide plan – an increase of 60% since 2011. Emergency department (ED) visits for mental health conditions among children and adolescents have also risen sharply. Individuals aged 14-18 have rates of ED visits for suicidal ideation that are higher than any other group and more than twice the national average. In 2021, the American Academy of Pediatrics, American Academy of Child and Adolescent Psychiatry, and Children's Hospital Association jointly declared a national emergency in child and adolescent mental health. The Medicaid program can play a critical role in addressing this mental health crisis. Medicaid is the largest national payer for mental health services and, in 2022, covered 50% of all children and adolescents in the U.S. Most of these children and adolescents receive care through Medicaid managed care organizations (MCOs), making MCOs a powerful mechanism for addressing the pediatric mental health crisis. However, little is known about how MCOs and state programs influence mental health care and outcomes for children and adolescents. The proposed mixed-methods study addresses this gap using the Transformed Medicaid Statistical Information System Analytic Files (TAF), a national Medicaid claims data set that includes comprehensive enrollment and service utilization claims and detailed information on MCO and providers. These quantitative analyses will be integrated with qualitative interviews with MCOs, Medicaid administrators, and other stakeholders. Our aims to seek (1) to characterize MCO structures using latent class analysis; (2) to assess associations between MCO structures and mental health access and utilization for adolescents with anxiety, depression, and suicide- related behaviors; (3) to test the implications of MCO exit on mental health access and utilization for adolescents; and (4) to test the impacts of acute adolescent mental health events on adult caregivers. The study will also be informed by close coordination with a deployment-focused advisory committee, which will assist in identifying feasible and scalable policies and processes that can improve access and outcomes related to child and adolescent mental health. In response to PAR-23-095, our proposed study will identify policies and factors that affect access, utilization, and outcomes, using large Medicaid claims datasets and leveraging an existing data management approach to advance our research.

NIH Research Projects · FY 2026 · 2024-07

Project Summary/Abstract We identified an association between lipoic acid and neural epidermal growth-factor like 1 (NELL1) membranous nephropathy (MN), an autoimmune kidney disease that causes urine protein loss (proteinuria), diffuse body swelling, and blood clots. If the condition is not detected early, NELL1 MN can lead to kidney failure. Lipoic acid is a widely available dietary supplement used for general wellness and a range of medical conditions including neuropathy and chronic pain. We have observed that 1/3 of NELL1 patients, mostly women (76%) with underlying autoimmunity (36%), report lipoic acid use. The long-term goal of our research is to investigate how lipoic acid promotes pathogenic autoimmune dysregulation so that we can detect early- stage disease in susceptible lipoic acid users, and deploy targeted treatment in those with NELL1 MN. Research suggests that approximately 70% of NELL1 MN patients have detectable anti-NELL1 antibodies in the blood, however we have an incomplete understanding about whether NELL1 antibodies can herald future disease. Furthermore, it is unclear how lipoic acid induces NELL1 antibody and the role of T cells in disease induction. Emerging data on T cell responses suggests a critical role for dysregulated CD4+ T regulatory cells and TH17 cells in another form of membranous nephropathy, and whether this dysregulated T cell signature is present in lipoic acid associated NELL1 MN is unknown. To begin to address these knowledge gaps, we first completed a study that demonstrates the feasibility of recruiting lipoic acid users, detecting circulating NELL1 antibody through Western blot, and examining peripheral T cell phenotype and function through flow cytometry. The feasibility study informs the design of this K23 career development award that will test the central hypothesis that circulating NELL1 antibodies and proinflammatory T cells predict proteinuria in susceptible lipoic acid users. We will test this hypothesis through two aims, 1) to determine whether NELL1 antibodies associate with proteinuria in lipoic acid users, and 2) to determine the phenotype and function of T cells in lipoic acid users who develop proteinuria. We will do this through an observational and longitudinal study where we will recruit 150 lipoic acid users at risk for NELL1 MN and measure circulating NELL1 antibodies through Western Blot, and collect peripheral blood mononuclear cells to examine T cell phenotype and function through flow cytometry, at pre-specified intervals over 2 years. This research has the potential to establish NELL1 antibody as a useful disease biomarker and demonstrate the role of T cells in disease induction, thereby unmasking possible treatment targets. The mentorship team of clinical and translational researchers with expertise in autoimmunity, lipoic acid, MN, and basic mechanisms of disease will support my training in autoimmunity, rare disease research, and leadership in science. This, combined with the ample resources at my institution for K awardees, will support my transition to research independence.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Currently, cardiovascular disease ranks as the leading cause of death in the United States. It is projected that by 2035, over 130 million adults in the U.S. will have some form of cardiovascular disease. Vascular graft technologies with patency rates equivalent to the clinical success of autologous grafting are lacking. Our goal is to utilize novel biomaterial surface modifications and develop nucleic acid delivery technology to improve host- biomaterial interactions with particular focus on the failure mechanism of neointimal hyperplasia resulting from smooth muscle cell proliferation and migration. Previous work by our team has optimized surface characteristics (physical topographies and biochemical modifications) to promote endothelialization without increasing thrombosis. However, the addition of controlled therapeutic nucleic acid delivery with this proposal will address intimal hyperplasia by directly altering smooth muscle cell phenotype. This work will develop the material surface delivery techniques to alter host-biomaterial responses, which occur after graft implantation. Our use of poly(vinyl alcohol) (PVA) biomaterials is integral to these studies because it is a highly modifiable material; however, these results can be readily translated to other modifiable off-the-shelf materials. Our aims are to: (1) develop nonviral delivery of miR145 with surface- and sustained-release via nanoparticles complexed to our novel PVA-CDI immobilized aminated fucoidan (PCAF) biomaterials, (2) characterize the effects of PCAF delivered miR145 on vascular cell responses, including endothelial cells, neutrophils, macrophages, and fibroblasts and (3) characterize the in vivo effects of PCAF-delivered miR145 on neointimal formation under healthy and diseased states. Using PCAF with previously established topographies and surface chemistries known to support endothelialization while maintaining hemocompatibility, we will utilize innovative gene delivery technologies in conjunction with our clinically relevant animal models. These strict models have the potential to improve translation by testing the material modifications in a whole, non-anticoagulated, flowing blood model of thrombosis and a clinically relevant end-to-side model of vascular grafting implantation. The deliverables for this project will include (1) efficient and sustainable delivery of miRNA-145 from our previously optimized PCAF to smooth muscle cells (SMCs), (2) induction of the SMC contractile phenotype without causing endothelial dysfunction, inflammation, or thrombosis, and (3) confirming a reduction of neointima formation in vivo without harmful “off-target” effects or harmful accumulation in organs. The potential significance of this translational project is the improved design of novel, functional biomaterial surfaces for cardiovascular applications. While we are focused on vascular graft applications in this proposal, the principles of surface controlled responses can be applied to multiple blood contacting devices. Our research will enable development and optimization of vascular biomaterials for use in humans.

NIH Research Projects · FY 2026 · 2024-07

Immune related adverse events (irAEs) secondary to checkpoint blockade inhibition (CBI) or other immune modulatory therapy for cancer, including head and neck cancers, are a significant problem given the increasing use of CBI and other immunomodulatory agents. It is therefore of critical importance to improve understanding and optimize treatment regimens for irAEs. Oral mucosal irAE toxicity is relatively understudied despite the potentially significant morbidity to patients. Understanding oral mucosal irAEs is an immediately accessible window into the autoimmune pathology caused by ICB. Lessons from oral mucosal irAEs thus can inform approaches to other irAEs. Our objective is to define T cell-based mechanisms of oral mucosal irAEs due to anti- PD-1 therapy. This work will also further our goal to identify predictive markers of oral mucosal irAEs through improved knowledge of their pathogenesis. Our central hypotheses are that regulatory T cell (Treg) plasticity is a major driver of oral mucosal irAEs, with IL-17A being a key cytokine driver of mucosal toxicity. These hypotheses build on prior work to define molecular signatures of T cells causative of muco-cutaneous eruptions. We will test these hypotheses using an innovative combination of approaches including analysis of human samples, mouse models that can generate similarly behaving irAEs, and generation and analysis of patient- derived organoids (PDOs). Our team of experts in immunology, immunotherapy, head and neck cancer, and bioinformatics will test this central hypothesis and achieve our objective via the following specific aims: 1) Determine functional significance of Treg and Th17 balance in oral mucosal irAEs. 2) Investigate mechanistically focused strategies to mitigate mucosal irAEs in murine models. 3) Validate therapeutic strategies to mitigate irAEs in patient-derived irAE organoid models. We will utilize our expertise in single cell RNA sequencing and PDOs to accomplish these aims. At the end of the project, the expected outcome is to better understand the pathophysiological mechanisms of oral mucosal irAEs and have identified strategies that can treat or modulate these, for future clinical trials. We will also be able to generate the mechanistic basis for future strategies to rapidly mitigate or even prevent these irAEs, which will ultimately benefit HNSCC and other cancer patients.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY This NIH Stage 1B study (9) will advance development of an intervention, STELLA-FTD (Support via Technology: Living and Learning with Advancing FTD), that is designed to address the needs of the approximately 50,000 family members in the US caring for someone with frontotemporal dementia (FTD). The STELLA-FTD intervention is framed by foundational research and informed by rehabilitative and nursing science. This study is important for two reasons. First, it addresses an important gap in care partner behavioral interventions. FTD is the most common dementia in persons under age 65, and one of the hallmark symptoms of FTD is distressing behaviors; yet, few behavior-focused interventions exist for FTD care partners. This study tests the preliminary efficacy of STELLA-FTD in reducing care partner burden related to the behavioral symptoms that come with FTD (e.g., disinhibition, apathy, agitation). Second this study tests the mechanism of action of the well-known ABC behavioral change technique. The ABC analytic approach involves identifying a behavior, then the behavior's activators and consequences. This approach is well-known in the caregiving world and research has demonstrated positive results in reducing care partner burden, but none has isolated the essential components of the ABC analytic approach. This limits future research and, importantly, scaling to community-based use. Upon completion of this study, should the hypotheses be proven, STELLA-FTD efficacy will be documented and the mechanism of action will be explained. This information will be the foundation for future NIH Stage 2 testing of STELLA-FTD. This 36-week randomized controlled (RCT), two-group, repeated measures trial with FTD family care partners will evaluate whether training in the use of the ABC analytic approach (test) reduces burden when compared to a dose-matched psychoeducational curriculum (control). This design allows for examination of the mechanism of action of STELLA-FTD by isolating the active component to determine its relationship with self-efficacy and care partner burden. The specific aims include testing of a) the intervention to assess preliminary efficacy, b) the mechanism of action and c) the mediating effect of self-efficacy on burden. In addition to testing STELLA-FTD, we will standardize materials and fidelity protocols to prepare the intervention for future NIH Stage 2 testing. We will test STELLA-FTD with 300 care partners. Both groups (n=150/group) will receive education about FTD and peer support. The test group will receive specific instruction in the ABC analytic approach, the control group will not. Efficacy will be assessed measuring burden between and within groups using Teri's Revised Memory and Behavior Problems Checklist (18). The key components of the mechanism of action will be isolated using the randomized design, with pre- and post-assessments of burden and self-efficacy of all participants. The data analysis will examine burden between and within groups, controlling for diagnosis type and demographic data.

NIH Research Projects · FY 2024 · 2024-07

Project Summary This proposal requests funds to be used to support travel and registration fees for postdoctoral fellows and graduate students to attend the 48th International Herpesvirus Workshop (IHW) in Portland, Oregon. The IHW is the major annual meeting for scientists studying all the herpesviruses and is very well established in its 48th year. This meeting has been very successful, in part, because of the enthusiastic participation of the world's leading herpesvirus researchers. The strength of the Workshop rests on the cross-fertilization that results from comparison of different herpesviruses, different approaches to key questions and on the support and participation of leading researchers in the field, most significantly including promising young investigators and students in training. Moreover, the forum is truly international, with broad-based world-wide attendance. The medical importance of this meeting is clearly indicated from the wide variety of diseases caused by the now- recognized eight human herpesviruses. These include skin and eye ulcerations (HSV-1), genital lesions (HSV- 2), meningitis and encephalitis (HSV-1 and HSV-2), infectious mononucleosis (EBV), chicken pox and shingles (VZV). CMV is a major cause of birth defects including mental retardation, blindness and deafness due to congenital transmission but also a significant opportunistic pathogen in AIDS patients and organ transplant recipients. In addition, CMV has been implicated as a pathogenic contributor in the development of atherosclerosis. Cancer has also been associated with herpesvirus infections. EBV is associated with Burkitt's lymphoma, other B cell neoplasias and nasopharyngeal carcinoma. The most recent human herpesvirus discovered (HHV-8 or KSHV) is associated with Kaposi's sarcoma in AIDS patients and other immunosuppressed persons in other groups. All of the herpesviruses persist for life and therefore pose significant problems in the treatment of immune-compromised individuals. Diseases caused by reactivation of most human herpesviruses are a significant cause of morbidity and mortality in various immune patient populations. Workshop sessions will take an interdisciplinary approach to the following topics: virus structure, mechanism of virus entry and cell-cell spread, membrane proteins, pathogenesis and latency, DNA replication, vaccination and the immune response, transcriptional control, regulation of gene expression, chemotherapeutic targets, and virus gene therapy.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Early initiation of antiretroviral therapy (ART) in perinatally infected infants diagnosed with HIV helps to reduce morbidity and mortality, but it is not sufficient to eradicate the virus. Children have a unique immune system that does not always respond to infection the same way as adults, and an increased understanding of the HIV-specific immune responses that persist in children after early ART is needed in order to inform development of immunotherapies that promote ART-free HIV viral control in children. In preliminary studies, we have shown that early initiation of ART before six months of age blunted the HIV- specific adaptive immune responses in children enrolled in the Thai HIV-NAT209 cohort. We were able to detect HIV-specific CD8+ T cells that persisted in these children as late as four years after ART initiation, but the frequency of CD8+ T cells that produced IFN in response to stimulation with HIV peptides was significantly lower in children compared to adults living with HIV on ART. As neonatal CD8+ T cells have impaired production of IFN, these standard assays may underestimate the frequency of HIV-specific memory CD8+ T cells differentiated during infancy. We have the unique opportunity to follow-up with children of the HIV-NAT209 cohort to determine if these HIV- specific immune responses persist through 6-10 years of ART, and further characterize their phenotype. We hypothesize that HIV-specific CD8+ T cells that persist after long-term antiretroviral therapy in children with early treated perinatal HIV will be less differentiated than those in early treated adults on long-term ART. To address this hypothesis, we will: (1) determine the magnitude, phenotype, and functionality of HIV-specific CD8+ T cells that persist in early treated children after long-term ART and (2) determine the T cell receptor repertoire and transcriptomic differences between HIV-specific CD8+ T cells in early treated children compared to adults after long-term ART. We will use unique functional cell-based assays and single-cell RNA-sequencing to analyze HIV- specific responses in these children and compare them to HIV-specific responses in early treated adults. Data from these studies will provide important information about the persistence and functionality of HIV- specific CD8+ T cell responses in early treated children, and will help to inform the development of pediatric targeted therapeutic interventions to promote ART-free viral control in children.

NIH Research Projects · FY 2026 · 2024-07

Project Abstract Every day, adult humans create and destroy tens of billions of cells. We know that imbalances in tissue homeostasis can lead to too many new cells, such as in tumorigenesis, and disruptions in dying cell clearance can cause inflammation, tissue damage, and autoimmune conditions. In species with the ability to regenerate from acute injury, waves of cell death are observed and are thought trigger appropriate levels of proliferation and inflammation. While decades of work have elucidated the pathways of cell death and corpse clearance during animal development, together called efferocytosis, relatively little is known about the dynamics of cell corpses in adult tissues or how efferocytosis machinery may function in the regeneration process. Major outstanding questions include: are cell corpses digested in place, or are they physically removed from tissues?; what cells do the engulfing in a given context?; and what happens to tissue turnover and regeneration when engulfment genes are disrupted? To uncover mechanisms of cell corpse clearance in adult regenerative animals, we utilize the complementary advantages of two model systems: planarians and zebrafish. The freshwater planarian, a flatworm from the phylum Platyhelminthes, has significant advantages to studying gene function in adult stem cell biology and regeneration in vivo. On the other hand, zebrafish are one of the best genetic models of regeneration and provide direct vertebrate relevance and the ability to watch cellular processes using transgenic lines. When combined, planarians and zebrafish can be used for gene discovery and genetic mechanisms of regeneration. In the current proposal, we developed a way to selectively kill off multiple cell types in planarians and can observe those cells transit from the periphery of the animal and be excreted through the gut. In our supporting data, we show that the gene engulfment and cell motility (ELMO) is required to clear cell corpses, but we do not understand how or through what cell types. Further, we developed a model of spinal cord regeneration using zebrafish, and show that multiple elmo genes are expressed in phagocytic immune cells as well as cell types in the spinal cord. Building on these data, in AIM 1, we will test how different cell types in planarians use the ELMO pathway to clear cell corpses in several contexts. In AIM 2, we will identify feedback signals from dying cells to stem cells and test how ELMO functions during regeneration. In AIM 3, we will use our zebrafish model of spinal cord regeneration to test how the ELMO pathway functions in cell corpse clearance and regenerative outcomes. Given that the ELMO pathway is highly conserved, our work will illuminate fundamental aspects of how defects in cell clearance influence regeneration outcomes and stem cell proliferation control, which will have high relevance to multiple human diseases.

NIH Research Projects · FY 2026 · 2024-07

Project Summary Ribosome biogenesis, a complex and coordinated cellular process for making the ribosome in the nucleolus, is essential for cell growth and proliferation. Defects in ribosome biogenesis are associated with a group of diseases called ribosomopathies. The goal of our research program is to understand how ribosome biogenesis is regulated during normal cell homeostasis and how it is deregulated in human diseases. Emerging evidence, including our own work, suggests that SUMOylation plays a critical role in ribosome biogenesis. We identified the ubiquitin-specific protease USP36 as a novel SUMO ligase that promotes the SUMOylation of snoRNP components Nop58, Nhp2, NOP56 and DKC1, the RNA exosome component EXOSC10, and the endonuclease Las1L, and plays a key role in multiple steps of rRNA processing. Meanwhile, deSUMOylation of the key nucleolar proteins by SENP3 is also essential for ribosome biogenesis. Thus, the proper nucleolar protein SUMOylation dynamics controlled by the coordinated action of USP36-mediated SUMOylation and SENP3-mediated deSUMOylation might be essential for ribosome biogenesis. However, there are several key gaps in understanding the SUMOylation dynamics in ribosome biogenesis, including the unclear mechanistic insights into the role of USP36 and SENP3, the limited scope of SUMO regulation of ribosome biogenesis identified, and the lack of dissection of the nucleolar SUMOylation dynamics. Our laboratory is well equipped in investigating ribosome biogenesis and protein posttranslational modifications. The goals of our research program for the next five years are to address these key gaps. We will investigate the mechanisms underlying the roles of USP36 and SENP3 using molecular imaging, structural biology, and molecular biology approaches. We will identify novel SUMOylation/deSUMOylation substrates of USP36 and SENP3 and dissect their roles in specific steps of ribosome biogenesis. We will then investigate the SUMO dynamics and its role in ribosome biogenesis and cell growth as well as the role of USP36 and SENP3 in cells in response to stress. Achieving these goals will provide critical insights into how USP36 and SENP3 properly coordinate to regulate nucleolar SUMOylation and ribosome biogenesis. The long-term vision of our research program is to understand how ribosome biogenesis is deregulated in various human diseases.

NIH Research Projects · FY 2024 · 2024-07

Project Summary Senescence and the senescence associated secretory phenotype (SASP) have long been implicated in driving tumorigenesis. However, the pro-tumorigenic mechanism(s) of senescence are still not fully understood, pointing to a need for better model systems to study the role of senescence in tumorigenesis. We recognized that a breast cancer model that used irradiation of cleared mammary fat pads followed by implantation of syngeneic Comma- D cells could be an ideal system to study senescence effects on tumorigenesis. Implantation of Comma-D alone does not result in outgrowth of tumors, but requires pre-irradiation of the gland for tumorigenesis. Similarly, treatment of the mammary fat pad with chemicals prior to implantation can also drive tumorigenesis. Both these types of treatments are also known to induce senescence, and indeed we found that irradiation induces massive senescence in the tissue. Thus, we hypothesize that tumorigenesis in this model system relies on induction of senescence to drive tumor formation, and that targeting the senescent cells or specific SASP factors will impair tumorigenesis. We propose to study this model system further to show that tumorigenesis requires induction of senescence in two specific aims. In the first aim, we will treat irradiated animals with senolytic drugs to remove senescent cells prior to implantation of SCp2 and other mammary cells, and compare tumor incidence in this group to a control arm that are treated with vehicle alone. Our preliminary data indicates that targeted removal of senescent cells with navitoclax blocks tumor formation. Extension of our studies will establish that senescence can drive early tumorigenesis in multiple models. In the second aim, we will examine how senescence drives pre-neoplastic cell growth, survival, and differentiation in vivo using multiplex immunofluorescence. We will perform RNAseq to identify SASP factors that are upregulated in response to irradiation. We will test these factors for their ability to enhance growth of SCp2 and human mammary epithelial cells (HMECs) in vitro. SASP factors that enhance growth and survival of these cells will be selected for targeting to determine their role in tumor formation. We will use small molecule inhibitors of receptors or interfering antibodies for SASP factors to determine how blocking these putative growth promoting factors impacts tumor formation. Successful completion of these studies will establish whether tumorigenesis in a luminal mammary cell implantation system is driven by senescence, will identify mechanisms by which senescence drives tumorigenesis, and will demonstrate the ability of senolytics or small molecule inhibitors targeting SASP factors to impair tumorigenesis. These represent the first steps in gaining mechanistic understanding of the role of senescence in early mammary tumorigenesis, with a long term goal of developing anti-senescence approaches for the prevention or treatment of human breast cancer.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Retinoblastoma (RB) is the most common primary intraocular malignancy of childhood. Although retinoblastoma is an aggressive cancer, early diagnosis and treatment may prevent visual impairment, and even improve mortality rates. This is particularly relevant for children at risk for retinoblastoma due to family history of the disease, which can be inherited in an autosomal dominant fashion. These children benefit fromea rly and frequent screening evaluations. Currently, RB diagnosis and management remains largely dependent upon ophthalmoscopic evaluation by expert clinicians. Ophthalmoscopy offers panretinal imaging, but suffers from a poor sensitivity to sub-millimeter (subclinical) tumors and a limited ability to monitor treatment response. Handheld optical coherence tomography (HH-OCT) offers the promise to address both of these needs, but technical deficiencies in imaging speed and axial range severely limit is clinica l utility. Recent innovations in circular-ranging methods, stretched-pulse mode-locked frequency comb lasers, and ultrawide handheld microscopes open new avenues for clinically meaningful deployments of HH-OCT in the management of Rb. In Aim 1, we will integrate these innovative technologies into a prototype instrument providing panretinal high- resolution three-dimensional imaging. In Aim 2, we will perform pilot imaging studies in sedated children undergoing RB screening. Study data will be used to assess the feasibility of panretinal imaging by HH-OCT in sedated children, to quantify RPE and ILM visibility as an image quality metric predictive of the instrument’s ability to detect subclinical tumors, and to confirm that the volume of all imaged tumors, regardless of size, can be calculated.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Gap junction channels are critical to vision by maintaining homeostasis in the lens and propagating electrical signals in the retina. Age-related stress and genetic mutations in connexins 46 and 50 (Cx46/50) in the lens and connexin 36 (Cx36) in the retina have been linked to cataracts, glaucoma, and retinopathies. Despite their vital importance to vision biology, we still lack effective pharmacological tools to elucidate the physiological and pathophysiological roles of gap junctions and their potential as therapeutic targets. Current small molecule modulators of gap junctions lack potency and isoform specificity, often interacting with and inhibiting other ion channels, eliciting unwanted off-target effects. Drug development has been slow in this field due to the lack of high-resolution structures of gap junctions in complex with a drug-like molecule. Recent advances in single particle Cryo-EM have enabled us to solve the structures of ion channels to near-atomic resolution and capture inhibitor-bound states. The Aims of this proposal will leverage these recent advances, in combination with fragment antigen-binding (Fab) technologies to deconvolute the mechanism of inhibitory action against native heteromeric gap junctions. To glean insight into the selectivity potential of gap junction inhibitors, this proposal will target the structural effects of mefloquine (MFQ) against the major gap junctions in the visual system. MFQ is one of the few drugs that exhibits gap junction selectivity against Cx50 and Cx36 isoforms. Remarkably, MFQ does not show appreciable binding to the highly related isoform, Cx46. In Aim 1, I will resolve the atomic structures of homomeric Cx36 and Cx50 in complex with MFQ. Comparative analysis to structures of Cx46 and Cx26 (no binding) and Cx43 (semi-inhibitory) will be used to identify interactions that are critical for MFQ selectivity and drive structure-guided mutation studies to validate the selectivity mechanism in vitro. Remarkably, MFQ inhibits native heteromeric gap junction channels composed of Cx46/50. In Aim 2, I will utilize high-affinity Fabs to identify co-assembly patterns of native mammalian lens gap junctions in the presence of MFQ to understand how this inhibition is achieved, and potentially elucidate the proposed cooperativity. Mechanistic insights garnered from these works will aid in designing the next generation of gap junction pharmacological probes to better understand the precise roles of gap junctions in vision health and disease.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Primary open angle glaucoma (POAG) is a leading cause of irreversible blindness, and elevated intraocular pressure (IOP) is the primary risk factor. By 2040, it is predicted that 112 million people worldwide will have glaucoma. The IOP is maintained through a dynamic balance between the production and drainage of the aqueous humor primarily via the trabecular meshwork (TM), juxtacanalicular connective tissue (JCT), and Schlemm's canal (SC) endothelium of the conventional outflow pathway. Outflow tissues are hyperviscoelastic (large-deformation and pressure-dependent) and dynamically interact with aqueous humor through an active, two-way, fluid-structure interaction coupling. While glaucoma affects the morphology and mechanical properties of the outflow tissues, the biomechanical and hydrodynamics states in glaucoma eyes are still largely unknown. Assessment of the dynamic biomechanical properties of the TM, JCT, and inner wall endothelial cells of SC with their basement membrane tissues in normal and disease will improve our understanding of the mechanisms of IOP regulation and the dynamic outflow resistance and could be a target for new diagnostic and therapeutic biomarkers in glaucoma. The goal of this project is to assess the biomechanical and hydrodynamic effects of different drugs and treatments on outflow facility modulation through an inverse finite element method coupled with an optimization algorithm and advanced OCT imaging data. In this project, the low-flow (LF) and high-flow (HF) regions in human donor eyes will be determined by confocal imaging of the FluoSpheres in the outflow pathway. Two-photon excitation microscopy will be used to measure the velocity of microbeads in the TM while the SC is negatively pressurized. The TM/JCT/SC complex in the LF and HF regions will be dissected and subject to tensile loading until failure. The elastic moduli of the normal and glaucomatous human donor eyes in the LF and HF regions will be calculated. A quadrant of the anterior segment from normal and glaucomatous human donor eyes in LF and HF regions will be positively and negatively pressurized and imaged using a visible-light green OCT. The TM, JCT, SC inner wall boundaries in a course of dynamic motion will be segmented using a deep neural network algorithm to reconstruct a finite element mesh of the TM, with the adjacent JCT and the SC inner wall. A mesh-free, beam-in-solid material-modeling algorithm will be used to distribute the collagen fibrils into the ECM of the TM and JCT. An inverse finite element method will be coupled with an optimization algorithm to characterize the ECM/collagen fibrils hyperviscoelastic mechanical properties in the LF and HF regions such that the TM, JCT, SC inner wall dynamic motion and OCT imaging data best match over time. 3D finite element model of the TM/JCT/SC complex will be reconstructed using the OCT images and will be subject to aqueous humor pressure elevation using fluid-structure interaction method. The biomechanical and hydrodynamics of the outflow pathway will be modeled and validated versus the digital volume correlation data (OCT images) and two-photon excitation microscopy/3D particle image velocimetry data.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Bipolar cells (BCs) are the first interneurons in the retina and are responsible for sampling photoreceptor output and distributing it to ON and OFF streams. Each BC subtype collects and transforms different salient components of the visual scene (luminance, frequency, contrast, etc.), yet the specific mechanisms different subtypes use to process this information into functional channels remain elusive. This proposal focuses on the CBC2 (Type 2 OFF cone bipolar cell). In addition to its conventional role processing photoreceptor output for downstream retinal ganglion cells, the CBC2 is the primary recipient of inhibitory crossover inhibition from AII amacrine cells and provides the majority of glutamatergic feedback onto AIIs, positioning it as an important mediator of bidirectional signaling between ON and OFF steams across the retina. This project will characterize distinctive and novel physiological features of CBC2s to determine their unique contribution to retinal function. My preliminary experiments have identified three particularly interesting features of CBC2s that have not been seen in ON-BCs. I hypothesize that CBC2s, and perhaps OFF-BCs in general, possess unique mechanisms that are likely fundamental to their role in the retina. This proposal will examine each feature through three primary aims. Aim 1 will use electrophysiology, pharmacology, and super-resolution immunofluorescence microscopy to identify and characterize a postsynaptic glutamate receptor in CBC2 dendrites that may augment conventional excitatory input from photoreceptors. Aim 2 will combine patch-clamp electrophysiology with time-resolved membrane capacitance measurements to examine the consequences of microdomain control of vesicle exocytosis on short-term plasticity and the voltage-dependence of exocytosis. Finally, Aim 3 will investigate reciprocal glycinergic feedback between CBC2s and AII amacrine cells and use Ca2+ imaging and computational modeling to create a biophysical model of glutamate release from CBC2s. Combined, these experiments represent the first comprehensive examination of the biophysical properties of the mammalian CBC2, a vital step in determining the role it plays in retinal function. While the von Gersdorff lab has a proven and published record in the realm of retinal biophysics, my proposal will require learning diverse and novel techniques, such as super- resolution microscopy and computational modeling, for which I have sought specialized training from outside my lab. I have carefully assembled a mentorship team to provide me with the technical expertise and dedicated guidance required to achieve my planned scientific aims and further my professional goal of a career in academic research.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY The over 6 million Americans with Alzheimer’s Disease and Related Dementias (ADRD) are particularly vulnerable to burdensome, low-quality, and costly care at the end of life. Research has highlighted the need to increase the use of palliative care in patients with ADRD as they have needs (e.g., symptom burden) equal to those of advanced cancer patients but are much less likely to receive palliative care. Hospice is a component of palliative care usually delivered at the very end of life. An innovative model of hospice delivery exists, termed concurrent care, where hospice is delivered alongside usual medical care capitalizing on the benefits of an integrated approach. Concurrent care seems effective at enhancing outcomes in patients with advanced cancer, however, a significant knowledge gap exists regarding the impact of concurrent care among persons with ADRD. Understanding the impact of concurrent care on patient and family outcomes can provide opportunities to improve the quality of end-of-life (EOL) care and to inform future health policy. The central hypothesis of this proposal is that modifiable organizational and clinician characteristics and care processes are associated with increased use of concurrent care and that concurrent care is associated with high-quality end-of-life care for persons with ADRD. We will address this hypothesis using a mixed-methods, concurrent triangulation study design using both quantitative and qualitative methods in the Veterans Affairs (VA) Healthcare System. Aim 1 will determine temporal trends in concurrent care and the patient, facility, and market characteristics (including regional patterns in care intensity and hospice resources) associated with increased use of concurrent care, Aim 2 will determine the association of concurrent care with patient and family-centered outcomes using a cross- temporal propensity score matching difference-in-difference approach, and Aim 3 will focus on family members of ADRD patients and their clinicians’ EOL care experiences using qualitative methods. The results of this proposal will be impactful because we will identify modifiable factors as targets for clinical and policy interventions to improve the quality of EOL care in ADRD. Our mixed-methods approach will allow us to better understand contextual factors that influence EOL care quality and the use of concurrent care to inform future palliative care and decision-support interventions. This study will provide important policy-relevant information on concurrent care outcomes relevant to the mission of the National Institutes of Aging to support research concerned with the quality and continuing care of patients with ADRD and their families.

NIH Research Projects · FY 2025 · 2024-07

SUMMARY It is now possible to generate nonhuman primates (NHPs) with precise germline genomic alterations, enabling researchers to generate highly refined models of human disease. Significant investments by our team at the Oregon National Primate Research Center (ONPRC) have greatly advanced NHP transgenic technology, resulting in the births of five healthy, genetically altered rhesus macaques (RM) since 2019. While these advancements are extremely promising, existing methods to create transgenic NHPs are too slow and expensive for most research applications. Innovative technologies are needed to accelerate creation of transgenic NHPs. Here we propose to adapt site-directed recombination, a proven technology used in rodent transgenics, to fundamentally advance NHP transgenic generation. We will generate founder animals with a small “docking site” integrated into the AAVS1 safe-harbor locus, which is a well-characterized genomic site that provides consistent expression across tissues. Gametes collected and banked from these animals will provide an “off- the shelf" system for subsequent gene editing and transgenic model creation. Embryos or cells from animals encoding this acceptor site can be injected/transfected with a donor vector that contains the desired genetic cargo. Site-directed recombination mediates integration of the cargo into AAVS1, with high on-target efficiency and few limitations in cargo size. This technology has the potential to revolutionize the non-human primate transgenic field.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Despite the potential for patients to make a full recovery after experiencing an out-of-hospital cardiac arrest (OHCA) due to ventricular fibrillation (VF) or pulseless ventricular tachycardia (VT), mortality remains near 75%. This higher mortality is partly because upwards of 50% of patients do not respond to initial defibrillation attempts. My career goal is to become an independent, federally funded physician-scientist leading a research program focused on improving outcomes for patients experiencing OHCA. My K23’s overall objectives are to evaluate if earlier antiarrhythmic administration is associated with improved survival after VF/VT OHCA, how the route of administration interacts with this association, and if changing the sequence of antiarrhythmics for VF/VT OHCA can reduce the time to antiarrhythmic therapy in a pilot feasibility trial. My central hypothesis is that early amiodarone administration in VF/VT OHCA can improve survival significantly compared to current guidelines. The rationale is that current cardiac arrest care guidelines for delaying antiarrhythmic therapy until after three shocks may reduce the efficacy of these agents, increase overall arrest duration, and reduce survival compared to an early antiarrhythmic approach. I will complete my overall objectives through three specific aims: (1) to assess the association between the timing and route of amiodarone administration in VF/VT OHCA and patient survival prospectively; (2) to measure the serum amiodarone concentration of VF/VT OHCA patients to compare the bioavailability of intraosseous routes to intravenous while accounting for time since drug administration; and (3) to perform a pilot, stepped-wedge cluster randomized trial evaluating a modified algorithm for VF/VT treatment to reduce the interval from arrest to amiodarone administration. These three aims will be accomplished over five years through seamless continuation of a pilot cohort, using the region’s highest volume cardiac arrest receiving hospital to collect serum drug concentrations, and using existing infrastructure to conduct a pilot cluster trial. Complementing these aims are five career development aims to facilitate training in (1) exception from informed consent randomized controlled trials; (2) population pharmacokinetic analytic techniques; (3) causal inference and multilevel modeling; (4) adaptive clinical trial methodology; and (5) the K-to-R transition. This proposal is significant as it focuses on improving care in the high-mortality condition of cardiac arrest, aligning with the NHLBI strategic objectives of levering data science and optimizing therapeutic strategies for improving health. The research is innovative because it uses a cohort that captures novel data elements, combining this with the use of serum drug concentrations to evaluate the efficacy of the intraosseous route to ultimately inform, in combination with pilot trial results, future studies investigating the optimal timing, route, and dose of antiarrhythmics in OHCA with downstream results that could change cardiac arrest care guidelines used by nearly every health care professional in the world.

NIH Research Projects · FY 2024 · 2024-07

SUMMARY Astrocytes are one of the major glial subtypes of the central nervous system (CNS), evolutionarily conserved from Drosophila to humans with regard to their morphology, molecular composition, and function. Although their morphological complexity is thought to be essential for astrocytes to perform their myriad roles in the CNS by physically contacting and interacting with diverse neural structures, including synapses and the vasculature, we only know of a handful of pathways that critically regulate astrocyte morphology. From an unbiased, forward genetic screen in Drosophila, we surprisingly identified dSarm and dWnk/Fray, both of which have been implicated in axon degeneration, as potential new regulators of astrocyte morphology. Preliminary findings in Drosophila show that dSarm/Sarm1 and dWnk/WNK work synergistically to regulate not only axon degeneration but also astrocyte morphology. Here, we propose to leverage the larval zebrafish system to ask whether the role of Sarm1 and WNK in regulating astrocyte development is evolutionarily conserved. In the axon degeneration pathway, NAD+ depletion is thought to be upstream of dSarm/Sarm1 and dWnk/WNK activation, and Axundead (Axed) to be downstream. Whereas the importance of NAD+ depletion upstream of dSarm/Sarm1 and dWnk/WNK has been shown across species, the functional ortholog of Axed is unknown in vertebrates. We therefore propose to determine if and how these known axon degeneration pathways up- and downstream of dSarm/Sarm1 and dWnk/WNK also function in vertebrate astrocytes. By harnessing the pharmacological and genetic toolkits available in larval zebrafish, we will interrogate the involvement of the NAD+ biosynthetic pathway in controlling process dynamics and morphology of astrocytes in vivo. We will additionally perform a targeted reverse genetic screen to identify the functional vertebrate ortholog of Axed in mediating Sarm1/WNK effects on astrocyte morphology. Together, this study will define new roles for key molecules in axon degeneration in the regulation of astrocyte morphogenesis and further our understanding of the Sarm1 signaling pathway in vivo.