University Of Washington

NIH Research Projects · FY 2026 · 2022-05

Project Summary It is estimated that around 40% of the adult population of the United States is obese and thus at increased risk for several chronic illnesses. Current weight loss strategies for obese people are often ineffective and come with serious neurological side effects. This proposal aims to determine whether new strategies to treat and prevent obesity can be developed from current insights into biological aging. The transcription factor C/EBP-β regulates the expression of genes involved in fat catabolism and fat stores mobilization. Preliminary observations suggest that pro-longevity interventions, such as mTORC1 inhibition, prevent diet-induced obesity in mice and activate C/EBP-β. The nucleoside-analogue reverse-transcriptase inhibitor (NRTI) adefovir dipivoxil (ADV) also prevents diet-induced obesity and activates C/EBP-β, though independently of mTORC1 inhibition. Building on these premises, Aim 1 tests the hypothesis that activation of hepatic C/EBP-β leads to increased energy expenditure and fat catabolism, with net negative effects on weight and fat stores. Using transgenics and pharmacological approaches, we will determine the role of hepatic C/EBP-β in lipid metabolism and homeostasis in the face of obesogenic challenges and morbid obesity. State-of-the-art techniques will be applied to measure the impact of different isoforms of C/EBP-β on energy balance, glucose homeostasis, and endocrine regulation of glucose and lipid metabolism. Aim 2 sets out to determine whether mitohormetic stresses can increase lipid metabolism through activation of C/EBP-β. Using ADV and other mitochondrial stressors, we will measure activation of mitohormetic pathways and their connection with increased hepatic oxidation of fatty acids and energy expenditure. This proposal will be carried out in an institution with strong research programs in both aging and obesity/diabetes biology. The candidate will receive state of the art training in techniques and analytical tools necessary to the completion of both aims, including indirect calorimetry, insulin and glucose tolerance testing, and big data analysis. The candidate will also acquire an in-depth background in energy and nutrient homeostasis. Altogether, the experiments and training proposed will allow the candidate to build an independent and successful research program applying insights from geroscience to understand and investigate nutrient homeostasis, energy balance, and related metabolic disorders.

NIH Research Projects · FY 2025 · 2022-05

Experimental and Computational Studies in Genetic Cardiomyopathies PI: Farid Moussavi-Harami Abstract Cardiomyopathies, including hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM), are an ideal venue for implementing precision medicine strategies. This is due to more routine use of genetic testing and the vast amount of knowledge regarding underlying biophysical mechanisms of sarcomeric variants, which contribute to both DCM and HCM. While the mechanisms of how sarcomeric variants cause cardiomyopathies is an active area of investigation, it is clear that they disrupt the finely tuned force-generation properties of cardiomyocytes. Many investigators have used a variety of biophysical and biochemical assays to study mechanism of sarcomeric variants and then scale these studies up to cells, tissues and animal models. These approaches are informative, but incremental and unable to asses many variants at once. Success in this area requires robust high-throughput assays with the ability for analysis of thousands of divergent variants at once. Our proposal will directly overcome limitations in the field by applying data analytics to biophysical simulations and experimental cardiac twitches. The fundamental hypothesis is that the principal features of cardiac twitches summarize the complex intra and inter-filament interactions of sarcomeric variants. Moreover, we can utilize these features for variant classification, predicting therapeutic response and identification of new therapeutics targets. Testing these hypotheses requires 1) large datasets of variants, 2) models that account for variant location and abundance in sarcomeres and 3) development and validation of data analytic methods. Biophysical simulations of sarcomeric variants can provide such datasets, but require validation in experimental systems. We will use a spatially explicit computational model of the sarcomere that can simulate how perturbations in sarcomere mechanochemistry change myocyte force generation. Simulated twitches will be generated, validated and used for predicting targeted therapeutics.

NIH Research Projects · FY 2026 · 2022-05

PROJECT SUMMARY Chronic pain affects up to one third of the United States population, and the reliance on opioids to treat chronic pain has contributed substantially to the opioid epidemic. Developing alternative pain therapies is critical to reducing the use of opioids, and the phyto-cannabinoid cannabidiol (CBD) is a promising candidate. We have shown that CBD has the ability to reduce chronic neuropathic pain-like responses in mice over 3 weeks. Terpenoids (eg beta-caryophyllene), also found in the cannabis plant, are a potential second class of pharmacologically active compounds in cannabis with possible analgesic benefits, although their pharmacodynamic properties in vitro and in vivo are poorly understood. Likewise, there is an incomplete understanding of how CBD and terpenoids produce anti-nociception, by themselves or combined (“entourage”), and their supraspinal neuropharmacological mechanisms of “pain control” remain unknown. The proposed experiments will first test whether specific combinations of CBD and terpenoids can produce short-term inflammatory and long-term neuropathic antinociception without tolerance. We will then establish the action of CBD and terpenoids in a critical brain nucleus for pain, the basolateral amygdala. Finally, we will determine the pharmacological and biochemical signaling profiles of CBD and terpenoids in vitro and in vivo. In Aim 1, we will use mixtures of CBD and terpenoids, investigator administered as well as in our gelatin self-administration model, and measure both consumption and pain scores after partial sciatic nerve ligation to determine if CBD and/or terpenoids provide analgesic benefit over protracted periods of pain. We will also verify that these mixtures are not inherently rewarding, which is critical for substance abuse liability. In Aim 2, we will test whether the amygdala is a critical brain circuit site for CBD/terpenoid analgesic action using single cell calcium imaging of amygdalar neurons during pain states, in parallel with local injections of CBD and terpenoids into the amygdala to produce analgesia. Further, we will knock out cannabinoid receptors and other putative sites of CBD/terpenoid action within the BLA to establish necessity of each in producing behavioral and physiological responses. In Aim 3, we will use in vitro and in vivo systems to determine the important biochemical features of CBD and terpenoid action at their putative receptor targets. We will measure cellular signaling activity via dynamic mass redistribution (DMR), MAP Kinase signaling, and the generation of reactive oxygen species. These aims will inform public health about the benefits and risks of long-term cannabidiol/terpenoid usage, as well as providing crucial mechanistic insight that will help develop and understand whether tailored medicinal cannabis approaches for chronic pain can be harnessed for therapeutic benefit.

NIH Research Projects · FY 2026 · 2022-04

Project Summary/Abstract The overall goal of our research is to elucidate the proximal molecular drivers and mechanisms of systemic lupus erythematosus (SLE) to enable the development of new and better therapies that target these processes but spare the broader immune system. This proposal will explore the role of the Long Interspersed Nuclear Element-1 (LINE-1, or L1) in type I interferon (IFN)-positive SLE. We have discovered that most SLE patients have high-titer autoantibodies against the first protein encoded by L1, ORF1p and that the titers of these autoantibodies correlate with disease activity as measured by SLEDAI score, complement consumption, presence of nephritis, anti- dsDNA and other autoantibodies, as well as with type I interferon gene induction. Our working hypothesis is that the biology of L1 is intimately connected with the pathogenesis of SLE by two mechanisms: i) driving autoimmunity to the virus-like L1 protein complexes that contain RNA and DNA; and ii) generating aberrant DNA by reverse-transcribing RNAs (including its own) and triggering DNA-sensor signaling to induce type I IFNs. AIM 1. To characterize the expression of L1 in SLE. The cell types that express the highest levels of L1 in SLE are the neutrophil and the low-density granulocyte. The L1-expressing neutrophils show signs of activation, and biomarkers of neutrophil NETosis are elevated in patient serum. By RNA-Seq we have identified one dominant (chr4q22.1) and six additional distinct human-specific L1 elements as the origins of the elevated transcripts and ORF1p protein. We will solidify these observations, detect and analyze their dependence on patient gender and disease parameters. We will examine factors that influence the transcription of these loci, and we will ask if stress granules containing ORF1p are released from neutrophils undergoing NETosis or other forms of cell death. These experiments will be complemented by asking if other proteins present in stress granules are also autoantigens in SLE. AIM 2. To elucidate the role of L1 ORF2p in type I IFN induction in SLE. Several recent papers in top journals have demonstrated a connection between L1 expression and the induction of IFN in cellular senescence, Crohn's disease, and in cancer cells. To determine how these findings apply to SLE, we will analyze L1-expressing neutrophils for activated DNA sensors, the signaling pathways from these sensors, and the type I IFN themselves. We will use an ultrasensitive method to quantitate ORF2p and then use inhibitors of the its RT activity to determine if it drives type I IFN production. Lastly, we will ask if silencing the DNA sensor DAI/ZBP1 blocks this IFN production.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY / ABSTRACT The goals of this K23 application are to support shared decision-making for older adults considering kidney transplant evaluation and to facilitate Dr. Catherine Butler's transition to independence as a physician scientist whose research focuses on making the transplant evaluation process more person-centered and equitable. Kidney transplant can be a valuable treatment option for the growing number of older adults with kidney failure, but because of their limited life expectancy and high burden of comorbidity, most will not receive a kidney. Further, the transplant evaluation itself can be time-consuming, burdensome, and risky, especially for older adults who are particularly vulnerable to the complications of aggressive screening and treatments. Our preliminary work suggests that patients often have little understanding of what to expect from the transplant evaluation and limited opportunity to shape their involvement. A process of shared decision-making could help older adults to navigate the complex trade-offs and uncertainties involved in the kidney transplant evaluation. We propose an integrated research agenda intended to strengthen shared decision-making around referral for kidney transplant among older adults. First, we will conduct an interview-based qualitative study to elicit the experiences and perspectives of older adults with kidney failure related to the transplant evaluation process along with their family members and clinicians (Aim 1). Second, we will describe the duration and likelihood of completing discrete steps in the transplant process for a large national cohort of patients with kidney failure (Aim 2). Finally, informed by these first two aims, we will adapt and pilot-test a communication tool to support shared decision-making around referral for transplant among older adults with kidney failure (Aim 3). During the award period, Dr. Butler will build upon her background in bioethics and early experience in qualitative and epidemiologic research to acquire new expertise in the use of applied qualitative methods, design and conduct of cohort studies, and development and testing of communication tools. This career development plan will be supported by the exceptional research environment at the University of Washington, including the Kidney Research Institute and the Seattle-Denver VA Center of Innovation. Dr. Butler's primary mentor, Dr. Ann O'Hare, is an internationally recognized physician scientist whose research focuses on complex care processes for older adults with kidney disease. Dr. Butler's co-mentors have complementary expertise in transplant epidemiology (Dr. Peter Reese) and communication tool development and testing (Dr. Margaret “Gretchen” Schwarze). The work proposed here will lay the foundation for a future trial to test the communication tool developed during the award period and, more broadly, will identify multiple avenues to improve upon the transplant evaluation process at both the individual- and systems-levels. Collectively, this work will position Dr. Butler to become a successful independent physician scientist whose research focuses on fostering a more person-centered and equitable approach to the kidney transplant evaluation process.

NIH Research Projects · FY 2026 · 2022-04

Opioid use disorder (OUD) is a prevalent and often life-threatening condition that can be effectively managed with medication-based treatment (MOUD). First-line therapies such as methadone and buprenorphine are known to reduce the risk of overdose and support long-term recovery. However, access to buprenorphine has not been consistent across all patient groups, raising questions about treatment delivery and system-level variability. Buprenorphine may offer advantages for many individuals, including ease of access and reduced treatment burden compared to methadone. During the 2020–2022 national public health emergency, changes in clinical policy allowed for greater flexibility in MOUD delivery, including remote initiation of buprenorphine. These changes created new opportunities to examine whether broader treatment access affected patterns of care for different segments of the population. If such policies yielded uneven benefits, they may have unintentionally reinforced existing gaps in treatment receipt. This project will use mixed methods to assess how variations in MOUD access and retention shifted before and after the implementation of emergency-era policy changes within the Veterans Health Administration (VHA), the largest provider of substance use care in the U.S. The study will focus on patient populations that have previously experienced limited access to buprenorphine, using national electronic health record (EHR) data to examine changes over time and across communities. Additionally, qualitative interviews will explore patient experiences with OUD care and perspectives on how temporary policy changes were implemented in clinical practice. Study aims include: (1) evaluating how medication receipt and retention varied following policy shifts for patients from different demographic groups; (2) examining the influence of community-level factors on treatment trends; and (3) capturing patient lived experiences with care during this transitional period. Sampling for interviews will ensure representation across key characteristics, including medication history and demographic background. Study findings will inform future MOUD policy and clinical implementation efforts aimed at improving treatment consistency and care quality across systems.

NIH Research Projects · FY 2026 · 2022-04

Alzheimer’s disease (AD) is one of the most common neurodegenerative diseases and has a high global health burden. While several AD genetic risk loci have been identified, the causal variant at these genome- wide association study (GWAS) sites is still unknown despite intensive sequencing efforts. Importantly, however, most genome-wide sequencing methods fail to accurately resolve tandem repeats, over 40 of which are key drivers of neurological disorders. We hypothesize that a substantial fraction of genetic contributions to AD come from variable number tandem repeats (VNTRs) that often contain internal repeat units 20 nucleotides or more and that until now have not been systematically assessed genome-wide. We have surmounted critical sequencing barriers by establishing state-of-the-art long-read sequencing techniques to amplify and characterize the exact nucleotide sequence across tandem repeats. We recently reported the identification of a tandem repeat in WDR7 that is associated with Amyotrophic Lateral Sclerosis (ALS). Using the pipeline that we established for tandem repeat length estimation and validation, we propose to characterize human-specific tandem repeat expansions in AD. Our exciting preliminary analysis reveals many examples of repeats with increased or decreased length in AD. Several of these significantly differentially expanded repeats are in GWAS loci previously implicated in AD. Remarkably, the lead SNP at these loci is associated with stepwise changes in repeat length in individuals heterozygous or homozygous for the risk allele, suggesting that VNTRs may even play a causal role at these loci. We also identify rare instances of massive expansions, and/or internal sequence differences that depend on disease state. Our goal is to assess tandem repeats prone to expansion to estimate VNTR length genome- wide in AD to identify the contribution of novel repeat expansions in disease. We leverage large cohorts of whole genome sequence data from the Alzheimer’s Disease Sequencing Project. The thousands of genomes available means we can detect VNTR expansions in Hispanic, African American and Non- Hispanic White individuals as well as sex-specific effects of VNTR expansions. We will then perform targeted long-read sequencing of AD DNA samples to resolve the complete sequence of tandem repeats and define the consequence of internal sequence variation to AD. Finally, we will work to establish mechanisms of pathogenesis of novel tandem repeat expansions in AD. Our proposed studies will establish a novel paradigm to interrogate the mechanism of repeat expansion and will reveal insight into novel genetic factors that cause or modulate risk for AD.

NIH Research Projects · FY 2026 · 2022-04

East and Southern Africa is home to 6.2% of the world’s population but includes 54% of all people living with HIV (PLWH). In this region, three out of five PLWH are women, and there is a particularly high burden of HIV amongst adolescent girls and young women (AGYW). Over half of African women use family planning (FP) services. Integration of HIV prevention and treatment with FP services holds promise for supporting progress toward the UNAIDS 95-95-95 targets for testing, treatment, and prevention. Nonetheless, integration of even basic HIV prevention and treatment services into FP clinics remains low and how best to integrate these services is still unknown. In a previous trial, the Systems Analysis and Improvement Approach (SAIA), was an effective implementation strategy for improving HIV counseling and testing in a small selection of FP clinics in Mombasa County, Kenya when delivered by research staff. SAIA incorporates a cascade analysis tool, sequential process flow mapping, and cycles of micro-intervention development, implementation, and assessment to improve a care cascade. More data is needed to understand if SAIA is effective for also improving linkage to HIV care and screening and linkage to pre-exposure prophylaxis (PrEP) in FP clinics when SAIA is delivered at scale by Kenyan public health workforce. The first objective of this study is to conduct a cluster-randomized trial evaluating the effectiveness of SAIA versus control (usual procedures with no specific intervention) for increasing HIV counseling, testing, linkage to HIV care, and screening and linkage to PrEP in new FP clients and new and returning AGYW clients. There will be a particular focus on the HIV prevention and treatment of AGYW in this study and any AGYW presenting for FP care will be prioritized. Quantitative and qualitative data will be analyzed using the RE-AIM framework to evaluate the program’s Reach, Effectiveness, Adoption, Implementation, and Maintenance. To understand how SAIA could be integrated into national Ministry of Health policies and programs, activity-based costing will be conducted to estimate the budget and program impacts of SAIA, scaled to a County level, from a Ministry of Health perspective. It is hypothesized that compared to control, SAIA will be effective at increasing HIV counseling, HIV testing, linkage to HIV care, and screening and linkage to PrEP for new FP clients and all new and returning AGYW FP clients when delivered at scale by Kenyan public health staff. The implementation evaluation, costing, and budget impact analysis will establish a road map for national-level implementation, positioning Kenya as a global leader in integrating FP/HIV services.

NIH Research Projects · FY 2025 · 2022-04

Abstract The GTPase RAS functions as a molecular “on/off” switch, existing both in GDP-bound (inactive) and GTP- bound forms (active). Despite functioning as a simple binary switch, RAS is capable of directing complex and diverse cellular processes, including proliferation, migration, survival, and T-cell development. Recent work suggests that the ability of RAS to play complex, often conflicting roles in diverse processes results from differences in cellular context and and/or subcellular localization of its signaling. We have developed a novel chemical genetic tool–called Chemically-Inducible Activator of RAS (CIAR)–to study the dynamics of the signaling networks that are mediated by RAS activity. CIAR allows rapid and dose-dependent activation of RAS signaling with a cell permeable small molecule. With CIAR, we propose to use targeted, quantitative phosphoproteomics and transcriptomics to study the fundamental dynamic behavior of RAS-driven signaling. Subcellularly-localized versions of CIAR will also be used to determine the effects of localized RAS activation on the dynamics of RAS-mediated signaling. Finally, we will explore positive and negative feedback within RAS-driven signaling pathways by selectively enhancing or inhibiting downstream signaling components.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT Ion channels are integral membrane proteins with gated transmembrane pores that conduct ions down their electrochemical gradients to transduce chemical, mechanical, and optical signals into electrical signals. In this proposal, we leverage a host of innovative tools to decipher allosteric gating mechanisms in two subfamilies of TRP ion channels, TRPV1-2 and TRPM2. TRP channels are famous for their multimodal gating whereby stimulus modalities as diverse as heat, cold, ions, lipids, nutrients, other proteins, and a variety of natural products (e.g., capsaicin, menthol) are allosterically integrated to determine the activity of a central ion- conducting pore. Each stimulus modality regulates the conformational energetics of a sensing module. The sensing modules in turn regulate the conformational energetics and conductance of the pore. The sensing modules may be coupled to the pore, to each other, or both. TRP channels provide an ideal system in which to decipher how allosteric conformational energetics produce protein function because they are regulated by many stimulus modalities, and we have significant understanding of the correspondence between their structural domains and sensing modules. The goal of this proposal is to measure, for the first time, the conformational energetics of TRP channel sensing domains and their coupling to the pore and to each other to solve pressing questions in TRP channel biology. Our long-term vision is to understand the general themes that underlie allosteric conformational transitions in ion channels. Our recent technical advances combining fluorescence lifetime imaging microscopy (FLIM) and patch- clamp electrophysiology to measure conformational energetics in the pore and a sensing module simultaneously promise rapid progress toward this goal.

NIH Research Projects · FY 2025 · 2022-04

Project Summary/Abstract Cardiac cell death by apoptosis and/or necrosis is a hallmark of cardiac ischemic injury, pathological remodeling, and end-stage heart failure. In contrast to apoptosis, the role of necrosis in the pathogenesis of heart disease has been largely understudied. Emerging evidence has identified several forms of “programmed necrosis”, such as death receptor-mediated necrosis (termed “necroptosis”), mitochondria-mediated necrosis, and oxidative stress-induced necrosis. How programmed necrosis is regulated in the heart remains largely unknown, and preventing necrosis is still an important challenge. Moreover, currently no molecular strategies are available to simultaneously target multiple cell death processes in heart disease. Here, our preliminary studies identified an unexpected role for CYLD (cylindromatosis), a lysine 63 (K63)-specific deubiquitinase, as a key regulator of multiple cell death pathways in cardiomyocytes, including apoptosis, necroptosis, and oxidative stress-induced necrosis. Intriguingly, CYLD expression was markedly upregulated in the heart following ischemic injury. Using Cyld knockout and transgenic mouse models, our preliminary data further show that ablation of CYLD attenuated, whereas overexpression of CYLD exacerbated, cardiac ischemic injury. Importantly, ablation of CYLD inhibited apoptosis, necroptosis, and necrosis in cardiomyocytes, whereas overexpression of CYLD showed the opposite effect. Mechanistically, our data reveal a K63-linked polyubiquitination (K63-Ub) dependent cell death signaling mechanism whereby CYLD controls the ubiquitination status and activity of three cell death regulators: TRAF2, TAK1 and AKT/PKB. Therefore, we hypothesize the deubiquitinase CYLD is a key regulator of multiple cell death pathways and a promising therapeutic target for cardiac ischemic injury and remodeling. Using genetic loss- and gain-of-function strategies, we will address two specific aims: Aim 1) To investigate the novel role of CYLD as a key regulator of myocardial cell death, ischemic injury, and remodeling using Cyld knockout and transgenic models and AAV9-shCYLD vectors. Aim 2) To define a CYLD-mediated, K63-Ub dependent cell death signaling network regulating apoptosis, necroptosis, and necrosis in cardiomyocytes. This project investigates a novel CYLD-mediated cell death signaling network in the heart and its functional relevance in cardiac ischemic injury and remodeling. Moreover, the proposed studies will define a K63-Ub dependent mechanism regulating apoptosis, necroptosis, and necrosis, which constitutes a new paradigm of cell death regulation. These studies also have important translational implications by providing new anti-cell death strategies, given our preliminary results revealing CYLD as a molecular target for multiple cell death processes.

NIH Research Projects · FY 2024 · 2022-04

Limited recovery of function after stroke remains a major problem for millions. Disability persists in many, especially when hand function is limited. Existing therapies are limited and many have difficulties with activities of daily living, even after rehabilitation. Electrical stimulation of the brain has been proposed and used in early studies to try and aid recovery. In animals, stimulation delivered to the brain at precise times may improve the effect of stimulation. By stimulating at the beginning of movements, or just after another part of the brain is active, recovery may be enhanced. We will use recent advances in implanted brain stimulator technology to place wires on the brain surface in patients who are recovering from stroke. We will stimulate the brain during therapy sessions. Healthier brain regions near the stroke area will be stimulated at the time of activity of brain regions right over the stroke. This will allow the two areas to strengthen their connection and improve the effect of therapy. We will measure motor function before and after these different stimulation patterns. This will establish the ability to perform these experiments safely, and show that patients can tolerate well these painless stimulation sessions. We will look at the strength of connections between brain regions and study the brain networks related to motor function. We believe these connections and networks will become stronger with brain stimulation. If successful, this preliminary study would offer a new form of treatment for stroke recovery.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY (OVERALL) The Vision of the Seattle Tuberculosis (TB) Research Advancement Center (SEA-TRAC) is to facilitate a decrease in the global burden of TB through training and multi-disciplinary research. SEA-TRAC will accomplish this by catalyzing new avenues of research and training a cadre of new investigators that will make a meaningful impact on the global epidemic. SEA-TRAC will operate from three world class institutions in Seattle: The University of Washington (UW), Seattle Children’s Research Institute, and Fred Hutchinson Cancer Research Center. We will leverage close partnerships with local non-academic entities, such as the Institute for Systems Biology, Public Health Seattle King County, and PATH as well as international partners, such as the Kenya Medical Research Institute. The Administrative Core will lead SEA-TRAC and assume responsibility for strategic planning and operational management as well as promoting an environment to foster dialogue that will lead to new collaborations and funding applications. The heart of SEA-TRAC will be the Development Core, which will oversee several new educational, training, and grant programs designed to foster career development of junior or senior investigators that are new to TB research. The Clinical and Translational Science Core will lead training and consulting in clinical research methodology and foster collaborative research with our international partners. The Basic Science Core will provide training for scientists new to working in the Biosafety Level 3 (BSL3) environment and lead training and consulting in advanced microbiology and immunology methods. Finally, the Data Sciences Core will leverage our local strengths in Biostatistics, Computational Biology, and Modeling to host community building events, such as TB ‘hackweeks,’ provide training to scientists that are new to data science, and offer consulting services for advanced research questions. Our Leadership Team is composed of faculty with a demonstrated commitment to training and mentorship that represent the diversity of scientific interests, tenure stage, gender, and ethnicity of our research community. We have secured institutional commitments totaling $861,420 or 29% of total direct costs that will allow us to support a diverse suite of programs immediately upon funding. SEA-TRAC will build on the strong foundation that has been established over the last five years by the UW Tuberculosis Research & Training Center (TRTC) to nucleate, strengthen, and expand the impact of our research. By the end of the funding period, we expect that SEA-TRAC will have achieved several quantifiable objectives and established itself as a flagship TRAC program, leading new initiatives through interactions with other TRACs and the NIH.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT Surgical site infection (SSI) is a major public health problem with devastating perioperative outcomes, affecting as many as 1 in 20 patients undergoing instrumented spine surgery. Procedures such as spinal fusion comprise the largest overall category of US spending on surgical care and are frequently performed for patients with pain or disability arising from a wide range of musculoskeletal conditions, such as congenital or degenerative disease, trauma, and neoplastic disorders. However, SSI rates in instrumented spine surgery are among the highest of any procedure involving a clean skin incision and they have not substantially decreased in decades. The development of more effective strategies for SSI prevention in spine surgery is significantly limited by a lack of fundamental understanding about the origins of causative bacteria, the basic pathogenesis of spinal wound infection, the microbiome of the back, and the role of antimicrobial resistance to surgical prophylaxis. We recently demonstrated that the microbiologic causes of spine SSI may vary by operative level and patient sex, and that most infections are resistant to the surgical antibiotic prophylaxis administered. The objectives of this K23 proposal are to build upon this foundation through training in translational microbiome sciences and by prospectively characterizing the role of the patient microbiome in spine SSI. The central hypothesis is that most spine SSIs arise from strains colonizing the patient prior to surgery (rather than acquired in the hospital environment) and that clinically actionable features of the preoperative patient microbiome strongly influence individual, modifiable risk. The long-term objective of this work is to use novel bacterial genomic techniques and large clinical datasets to identify the fundamental mechanisms by which spinal wound infection occurs, enabling the development of more effective prevention strategies. The specific aims of this proposal are to: 1) define preoperative bacterial genetic features of Staphylococcus aureus associated with spine SSI, allowing development of improved screening and decolonization measures, 2) identify sources of endogenous gram-negative spine SSI and associated resistance to surgical antibiotic prophylaxis, enabling prevention strategies for this important class of infection that may disproportionately affect specific groups (e.g., women undergoing lumbosacral procedures), and 3) determine clinical risk factors for resistance to surgical antibiotic prophylaxis in spine surgery to inform tailored approaches to selection of antibiotic prophylaxis in spine surgery for diverse patient populations. These research activities are closely aligned with my career development plans. Through the team of expert mentors assembled from anesthesiology, surgery, and molecular microbiology, I will receive training in cutting-edge translational microbiome research and career development toward independence. This award will prepare me for my first R01 submission on tailored SSI prevention tools/strategies for clinical use in spine surgery and for an independent translational research career in perioperative infection prevention.

NIH Research Projects · FY 2026 · 2022-03

Project Summary Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a recently emergent, currently pandemic virus and etiological agent of Coronavirus Induced Disease-19 (COVID-19). Despite a flood of scientific investigation, critical gaps remain in our understanding of the basic cellular processes that facilitate replication of coronaviruses, including SARS-CoV-2, and contribute to the pathogenesis of severe disease. Our preliminary data demonstrate that IRE1α, a component of the cellular response to endoplasmic reticulum (ER) stress, is required for SARS-CoV-2 replication and inflammatory cytokine responses. However, the stage(s) of the viral life cycle and downstream cellular pathways that mediate these effects remain completely unknown. ER stress and IRE1α activation are well-associated with conditions such as obesity, diabetes, hypertension, and aging, all of which are risk factors for severe manifestations of COVID-19. We hypothesize that comorbidity-associated ER stress primes both exuberant viral replication and pathogenic inflammatory cytokine production via IRE1α. This project leverages our unique ability to test this hypothesis using cell culture infection models, as well as specimens from patients with COVID-19. IRE1α inhibitors are under evaluation for treatment of non-infectious human diseases, and we propose that this project will provide preclinical evidence for the novel application of these drugs to treat infections with SARS-CoV-2 and potentially other human coronaviruses. The experiments outlined in this proposal will determine the molecular mechanism(s) by which IRE1α supports SARS-CoV-2 infection. IRE1α is a nuclease which initiates nonconventional splicing of XBP1 mRNA, which encodes a pleiotropic transcription factor. IRE1α also targets other specific RNAs leading to their degradation. We will determine whether the requirement for IRE1α is XBP1-dependent or -independent and dissect the downstream cellular processes that facilitate SARS-CoV-2 replication and inflammatory cytokine responses (Aim 1). We will systematically identify stage(s) of the SARS-CoV-2 life cycle that require IRE1α (Aim 2). We predict that IRE1α most likely supports biogenesis of ER-derived viral replication platforms, and will focus experiments on this hypothesis. We predict that IRE1α represents a target for dual anti-viral and anti- inflammatory therapy and will test this in mouse models of SARS-CoV-2 infection (Aim 3). Finally, we will determine whether IRE1α activation occurs during human infection and ER stress is a prognostic marker for severe COVID-19. Together, the results of this project will reveal basic cellular processes occurring during coronavirus infection and host factors critical for the pathogenesis of COVID-19. .

NIH Research Projects · FY 2026 · 2022-03

Project Summary Parasitic worms (helminths) currently infect 1-2 billion humans and are the cause of widespread morbidity. Helminth infections of livestock result in large economic losses. Meanwhile, the incidence of allergies is increasing globally, but particularly in regions where helminths are absent. Although seemingly unrelated, helminths and allergens are immunologically linked as they both induce a type 2 immune response. The activation and regulation of type 2 immunity remain incompletely understood, especially in the small intestine (SI). We recently discovered that rare chemosensory epithelial tuft cells are required for anti-helminth immunity in the SI. Tuft cells activate group 2 innate lymphoid cells (ILC2s), which in turn secrete canonical type 2 cytokines to coordinate hallmarks of type 2 immunity, such as eosinophilia and tissue remodeling. Specifically, IL-13 signaling in epithelial progenitors biases their lineage commitment towards tuft cells, leading to tuft cell hyperplasia during helminth infection and in mice colonized with Tritrichomonas protists. This tuft-ILC2 circuit is activated when tuft cells sense succinate secreted by Tritrichomonas, but the ligand and receptor mediating helminth sensing remain unknown. Additionally, while the cells and intercellular signals of the tuft-ILC2 circuit are emerging, the intracellular signaling pathways that regulate the circuit remain completely unknown, especially within the epithelium. In this proposal, we examine how POU2F3 is regulated to induce tuft cell hyperplasia (Aim 1), test the function of KIT signaling in tuft cells (Aim 2), and characterize novel tuft cell ligands and the intracellular signals they activate (Aim 3). In addition to helminth infection, the tuft-ILC2 circuit has recently been implicated in enteric virus infection and ileitis induced by bacterial dysbiosis. Therefore, the regulatory mechanisms uncovered by this project promise to identify therapeutic targets for the treatment of numerous SI pathologies.

NIH Research Projects · FY 2026 · 2022-02

Project summary Detection and diagnosis of smaller and earlier-stage cancers significantly improves a patient's chances of survival. Positron emission tomography (PET) imaging using fluorine 18–fluorodeoxyglucose (FDG-PET) provides a functional or metabolic assessment of normal versus cancerous tissues, and since 2000 has been widely used clinically for the detection and diagnosis of many cancers. Studies over a decade ago by our group and others had shown that it was feasible to both measure and improve the detection ability of FDG-PET imaging for cancer by adjusting acquisition and image reconstruction parameters. This could be done systematically by evaluating the effect on observer models that replicated human performance (i.e. radiologists or nuclear medicine physicians). At the time, however, it is challenging to understand how this varied across systems with different resolutions, sensitivities, and reconstruction algorithms, or if they were operated differently across imaging sites. Over the last decade there have been dramatic improvements in scanner resolution, sensitivity, and reconstruction algorithms, as well as the routine adoption of time-of-flight PET imaging. In parallel there has been an improved understanding and adoption of model observers, as well as pathways for adoption or harmonization of methods across multiple PET manufacturers and imaging sites. Most recently there has been the development of machine intelligence algorithms, such deep neural networks, for both image reconstruction and image analysis, which have the potential to improve performance. We are proposing to take advantage of these developments to characterize, optimize, and harmonize cancer detection with PET imaging. The three specific aims are: (1) Develop methods for characterization (i.e. measurement) of detection performance for FDG PET imaging. (2) Using a model system calibrated to a modern physical system we will then determine how to optimize cancer detection as a function of acquisition and image reconstruction parameters. (3) Finally we will develop a platform-independent (vendor agnostic) standard that can be applied across systems and imaging sites. This will lead to a roadmap for multi-site and multi-vendor implementation approaches that optimizing cancer detectability and thus improved patient outcomes.

NIH Research Projects · FY 2025 · 2022-02

SUMMARY: The purpose of this K01 proposal is twofold: 1) to identify early protein biomarkers in Alzheimer’s disease (AD) and cognitive decline; and 2) to provide Alison Fohner, PhD with the mentorship and resources to pursue an independent research career using multi-omic and longitudinal data to improve prevention and treatment of AD. Every year, 500,000 people are diagnosed with AD in the US. AD has no cure; and between 2002 and 2012, nearly every clinical trial for new therapeutics failed. Plasma proteins are easily measured in routine care, and protein levels may reflect underlying pathology. New technology that rapidly assays thousands of proteins in large samples promises to improve protein biomarker discovery, and could lead to new strategies for early risk stratification and for novel therapeutics. This proposal leverages the extensive existing data from the Cardiovascular Health Study (CHS), an NHLBI-funded prospective population-based cohort study of 5888 White and African American men and women recruited in the early 1990s. Available CHS data include plasma protein level data on 1300 proteins, whole genome sequencing data, yearly cognitive assessments covering different functional domains, diagnostic information on AD, and stored biospecimens. This proposal aims 1) to identify plasma proteins associated with time-to-incident AD and with rate of cognitive decline; 2) to assess genetic evidence for and against causal roles of high-signal proteins in AD and cognitive decline; and 3) to estimate the association of plasma p-tau181 concentrations, a biomarker of AD pathology, with subsequent clinical diagnosis of AD and cognitive decline. This proposed research will not only advance our understanding of AD pathology, but may also identify important clinical biomarkers and therapeutic targets for AD. Dr. Fohner is an Assistant Professor in the Department of Epidemiology at the University of Washington. With her background in genomics and high-dimensional data analysis, Dr. Fohner is well-positioned to pursue the Aims of this proposal. She has composed an experienced and collaborative mentorship team, and has developed an innovative training plan that will help her achieve research independence. The research and training plans in this proposal will prepare Dr. Fohner to successfully compete for future R01 funding by enabling her to build research skills and domain expertise, to learn new analytical techniques, to forge productive collaborations, and to generate preliminary data. In summary, with the support of this K01 award, Dr. Fohner can launch a successful career developing strategies to predict, prevent, and treat AD.

NIH Research Projects · FY 2026 · 2022-02

ABSTRACT Mammographic density, which describes the proportions of epithelial and stromal vs. adipose tissue in the breast, is a strong risk factor for breast cancer. Many of known breast cancer risk factors are also associated with mammographic density, suggesting that mammographic density is an important intermediate phenotype to study. Indeed, as a continuous and more precise outcome associated with all breast cancer subtypes, mammographic density has proven an important surrogate marker for breast cancer, and it has been argued that mammographic density is a more meaningful biological outcome to study. Breast cancer is the most common cancer and the leading cancer-related cause of death among Hispanic women living in the US. A recent study estimated that among Hispanic women, breast density accounted for 35% of premenopausal and 13% of postmenopausal breast cancers. Yet, an overwhelming majority of mammographic density research to date has been conducted among non-Hispanic Whites, leaving a substantial gap in our understanding of mammographic density and its determinants across racial/ethnic groups. We propose to conduct the largest epidemiological study of mammographic density among Hispanic women living in the US to date. We will first establish a repository of 3,200 mammograms within the multi-site US Hispanic Community Health Study/Study of Latinos (HCHS/SOL) cohort and obtain estimates of dense area, non-dense area and percent density phenotypes (Aim 1). We will then assess if sociocultural, reproductive and adiposity-related factors are associated with mammographic density phenotypes in a total sample size of 6,347 US Hispanic women (Aim 2), leveraging rich lifestyle and genetic data in HCHS/SOL together with already existing mammographic density, lifestyle and genetic data from the Mountain Park Health Center (MPHC) Mammography and Latinas LEarning About Density (LLEAD) studies, which include 3,147 US Hispanic women. Finally, we will conduct genetic association studies of mammographic density phenotypes, including genome- wide association studies (GWAS) in 3,300 women for whom we have GWAS and mammographic density data. Novel loci will be replicated in an independent population of 2,000 Hispanic women, and assessed for their association with breast cancer risk in a Hispanic GWAS of 4,500 cases and 9,000 controls (Aim 3). HCHS/SOL represents multiple Hispanic background groups (Dominican, Central American, Cuban, Mexican, Puerto Rican, South American), reflecting the diversity of Hispanics living in the US. Our mammogram repository will constitute the largest, most comprehensive collection of mammograms in a Hispanic population with already collected rich lifestyle and genetic data. Numerous future projects would build on this one-of-a-kind resource including collection of longitudinal mammograms to study change in density with time. With the growing number of Hispanics living in the US, we need to expand our epidemiological research into risk factors of common diseases, such as breast cancer, to achieve equal care across racial/ethnic groups.

NIH Research Projects · FY 2026 · 2022-02

Project Summary The International Registry of Werner Syndrome recruits cases of Werner syndrome (WS) and a range of other segmental progeroid syndromes from around the world with the goal of elucidating underlying mechanisms of accelerated aging. Detailed clinical information, the results of genetic analyses, and biological specimens are made available to a wide range of qualified geroscientists. In this application, we propose to extend our previous studies to include systematic genome-wide searches for the genetic variants responsible for the 78 progeroid cases that we have so far been unable to genetically characterize. An additional important extension of our research agenda is to initiate translational research that can lead to potential therapeutic agents. We will employ a combination of next generation sequencings, array CGH, and Sanger sequencing, followed by confirmatory Western analysis and quantitative PCR. These approaches have successfully identified novel pathogenic variants of WRN (a DNA helicase), LMNA (a component of nuclear structure), POLD1 (DNA polymerase delta), SPRTN (recruiter of DNA polymerase), ERCC4 (nucleotide excision repair), CTC1 (telomere replication), MDM2 (an inhibitor of P53) and SAMHD1 (regulation of dNTP pools). These loci highlight major roles for genome instability, now widely accepted as one of the hallmarks of aging. We also made progress in the identification of disease mutations that suggest other mechanisms of accelerated aging, such as BSCL2 (lipid droplet formation) and SMAD4 (intracellular signaling of a component of SASP, TGFβ). As indicated above, we will pursue translational research with the potential for the development of ameliorative therapies for our progeroid patients. Based on our previous studies, our collaborator, Dr. Yokote Koutaro at the Japanese Werner Consortium, is evaluating the efficacy of an NAD intermediate and an mTOR inhibitor, metformin, in WS patients. We shall begin independent studies of the effects of suppressors of chronic inflammation, namely Janus kinase (JAK) inhibitors, in cultures from WS patients and controls. This effort was motivated by our findings that SMAD4 mutant fibroblasts exhibited increased accumulation of DNA damage and that WRN mutant fibroblasts showed elevated SMAD4 expressions and dramatically higher levels of SASP compared to control cells, suggesting that a synergy of persistent DNA damage and inflammation may be one of the common key mechanisms leading to the accelerated aging. Concordant experiments will explore the effects of these novel therapeutic targets with high throughput screening of cell cultures from patients with other progeroid syndromes. An initial small scale experiment involving siRNA screening has revealed that siRNAs and drugs that alter the intranuclear dNTP concentration are able to modulate the cellular disease phenotypes of POLD1 mutants. Larger scale siRNA screening will be employed to identify additional novel relevant target pathways as well as previously unknown functional interactions of progeroid genetic loci.

NIH Research Projects · FY 2026 · 2022-02

ABSTRACT: Lewy body dementia is an Alzheimer’s disease–related dementia that affects more than one million Americans. The strongest risk factor for this devastating neurodegenerative disease is mutation of the GBA gene, which encodes the lysosomal enzyme glucocerebrosidase (GCase). GBA mutations can impair the ability of GCase to break down its substrate, the glycosphingolipid glucosylceramide (GlcCer), but it is not clear how defects in GlcCer breakdown increase the risk of Alzheimer’s disease–related dementias. To study this problem, we created a fly lacking the Drosophila ortholog of GBA, Gba1b, as a model of GCase deficiency. Our Gba1b mutant shows accumulation of GlcCer and recapitulates features of Alzheimer’s disease–related dementias including neurodegeneration, brain protein aggregates, and age-related cognitive decline. We now propose to use the Gba1b mutant to test a novel model of neurodegeneration associated with GCase deficiency, in which GlcCer accumulation leads to neuroinflammation via changes in extracellular vesicles (EVs). In earlier work, we found that Gba1b mutants had increased abundance and turnover of EV proteins, and that genetically suppressing neuronal EV production ameliorated mutant phenotypes. More recently, RNA-Seq experiments revealed that Gba1b mutants had marked innate immune activation. A followup RNAi screen of major innate immune pathways found that neuronal knockdown of two p38 MAPK pathway components, the transcription factor Atf-2 and its upstream kinase licorne, also suppressed Gba1b mutant phenotypes. Based on these and other findings, we hypothesize that immune-mediated neurodegeneration in Gba1b mutants is a process driven by excess GlcCer in two separate roles. Specifically, we hypothesize that excess GlcCer in neurons triggers ligand-independent receptor tyrosine kinase activity and p38 MAPK signaling, stimulating EV release; GlcCer in the released EVs then causes glia to secrete immune effector proteins, leading to protein aggregation and neurodegeneration. We propose three aims to address these hypotheses. The first will delineate the intraneuronal signaling pathway that leads to abnormal EV release; the second will investigate the nature of the EV alterations; the third will determine how those EVs trigger immune effector secretion from glia. Given the abundant evidence for neuroinflammation in Alzheimer’s disease and related dementias, we anticipate that our work will have broad medical significance.

NIH Research Projects · FY 2026 · 2022-02

Patient cost burden (PCB) for healthcare, as influenced by the rapidly changing health policy landscape, is a common patient-reported outcome for uninsured individuals and enrollees in high deductible health plans (HDHPs, 30% of employer-sponsored plans). In these contexts, PCB has important effects on adherence to medications that help manage cardiovascular and metabolic health (CMH). The management of CMH, including type 2 diabetes mellitus (T2DM), is an important part of healthy aging, and affects risk of Alzheimer's disease and related dementias (ADRD). In order to understand the long-term consequences of today's PCB on the healthy aging and ADRD risk of tomorrow, it is critical to develop methods and models that can simulate the complicated interplay between PCB, medication adherence, and ADRD. The proposed work in this application builds upon my experience investigating effects of insurance design and pharmacoepidemiology of ADRD, and enhances it by adding training in areas that are critical to achieving my long-term career goal: to become a leading independent investigator of the relationships between health policies and the burden of chronic diseases. The training of this award includes development and application of simulation models of healthcare interventions, policy and stakeholder engagement, T2DM and ADRD, geriatric research and care, and leadership. The newly acquired skills and knowledge obtained are necessary to conduct the proposed research, which aims to examine the effects of PCB on management of T2DM, and simulating how health policies that influence PCB affect the burden of ADRD. With mentorship from established leaders in simulation modeling, geriatrics, health economics, health policy, ADRD, and T2DM, I will accomplish the following specific aims: 1) Simulate long-term effects of PCB on T2DM management and progression using the Real-World Progression in Diabetes Model (RAPIDS). PCB will be examined in the context of uninsurance and HDHPs, and we will simulate how effects of PCB on T2DM management translate into long-term CMH outcomes in RAPIDS, a validated model of T2DM progression. 2) Build a new simulation model using a near-elderly/elderly lifetime perspective to connect PCB, T2DM progression, and ADRD incidence and burden. The new Healthcare Access Today and Healthy Aging Tomorrow (HATHAT) Model will connect the RAPIDS Model to the validated Future Elderly Model (FEM), in order to comprehensively relate PCB, T2DM treatments, micro- and macro- vascular events, and biomarkers to the future burden of ADRD. 3) Evaluate long-term consequences of highly relevant state and national policies on healthy aging in the HATHAT Model. Policy experts will advise on the most relevant and impactful policies to evaluate, and simulations will delineate effects along the PCB-->Treatment Use-->CMH-->ADRD nexus. Success of this project can lead to future models that examine health policies yet to be conceived, in a range of disease areas. This will leave me well-positioned to lead an independent research program that influences future policy by identifying how PCB and access to care relate to long-term healthy aging and ADRD.

NIH Research Projects · FY 2026 · 2022-01

Project Summary/Abstract Checkpoint blockade has revolutionized the field of cancer immunotherapy treatment, but many tumors remain unresponsive due to lack of effector T cell infiltration and activation in the tumor microenvironment. Innate immune priming of these “cold” tumors has emerged as a therapeutic strategy for increasing the efficacy of checkpoint blockade through stimulated type I interferon (IFN) production and downstream adaptive response. Recent efforts and ongoing clinical trials have focused on activation of the STING-dependent antiviral pathway to promote IFN production in the tumor microenvironment. However, many tumors downregulate STING signaling, and the therapeutic effects of STING agonists are thought to be mediated by their effects on tumor- infiltrating host myeloid cells. Our lab recently discovered that the DNA damage sensor DNA-PK triggers a STING-independent DNA sensing pathway (SIDSP) in human cells that potently activates IFN production in response to foreign DNA. We have developed synthetic superagonists of DNA-PK-dependent antiviral immunity that trigger potent antiviral responses in human melanoma cells that are unresponsive to STING agonists. We hypothesize that activation of the SIDSP within tumor cells will provide a unique signal to enhance inflammation within tumors and will stimulate potent immune responses. The goal of this proposal is to assess DNA-PK-SIDSP activation as a therapeutic strategy in human cancer. We will determine how DNA-PK directs distinct outcomes to DNA damage versus foreign DNA, and we will evaluate the therapeutic potential of triggering the DNA-PK-SIDSP in human tumors, in vitro and in vivo using cutting edge humanized mouse models. Our studies will uncover fundamental new aspects of the biology of the SIDSP, together with the first pre-clinical evaluation of DNA-PK activation as a novel cancer immunotherapy.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT Our goal is to reduce diagnostic and treatment errors, improve survival, and increase the value of care for lung cancer patients by improving our ability to select patients who benefit from a pretreatment lymph node biopsy. Accurately determining whether cancer has spread to lymph nodes and the extent of spread (a process called nodal staging) is critical for appropriate treatment selection. Understaging can lead to omission of chemotherapy or unnecessary surgery. Overstaging can lead to unnecessary chemotherapy and omission of surgery. Diagnostic and treatment errors negatively impact survival. These errors commonly occur when using imaging alone for nodal staging. A biopsy can reduce the chances of error, but it can also result in rare, life- threatening adverse events. Each biopsy costs ~$5,000. Practice guidelines recommend selectively performing a biopsy when imaging findings suggest nodal disease. However, national biopsy rates are less than half of what they should be. Moreover, there is 25-fold facility-level variability not explained by access to care, case- mix, or clinician or facility characteristics. These findings, along with the low levels of evidence underlying guideline recommendations, suggest true clinical and scientific uncertainty over the indications for lymph node biopsy. We conducted a pilot study to better understand how well guideline recommendations select patients for biopsy and learned that guideline-concordant nodal staging selects all patients with true nodal disease for biopsy and two-thirds of patients without true nodal disease for biopsy. Additionally, we developed and validated an alternative risk-based nodal staging strategy that uses a prediction model to stratify and select patients for lymph node biopsy. Preliminary data show that it identifies nearly all patients with true nodal disease for biopsy but selects fewer patients without nodal true nodal disease for biopsy. However, the relationship between selection strategies for lymph node biopsy and patient outcomes remains unknown. We hypothesize that guideline-concordant nodal staging is associated with higher 5-year survival rates compared with guideline-discordant nodal staging (Aim I) and that risk-based nodal staging is equivalent to guideline- concordant nodal staging in terms of survival but superior in terms of lower biopsy-related adverse events and healthcare expenditures (Aim II). Testing these hypotheses will require ~4,000 patients; therefore, a trial is not feasible at this time. We will create a novel cohort of lung cancer patients using the Cancer Research Network infrastructure to conduct Aim I using an observational, comparative-effectiveness study design with advanced regression techniques and machine learning to minimize confounding. Additionally, we will use patient-level data from this cohort as model inputs in a comparative-effectiveness simulation model that we will develop to conduct Aim II. Findings from this study will lead to: 1) developing and testing implementation strategies designed to increase guideline-concordant nodal staging, 2) alternative guideline recommendations for nodal staging, and/or 3) justifying trials comparing outcomes between different nodal staging strategies.

NIH Research Projects · FY 2025 · 2022-01

We recently showed that serum levels of apolipoprotein C3 (APOC3) predicted incident cardiovascular disease (CVD) in CACTI, a prospective study of subjects with type 1 diabetes mellitus (T1DM). In complementary mechanistic studies, we found that reducing APOC3 levels with an antisense oligonucleotide (ASO) prevented lesion progression in a mouse model of T1DM and that apolipoprotein B (APOB)-containing lipoproteins were driving accelerated diabetic atherosclerosis. These observations are important because they strongly support the proposal that APOC3 promotes atherosclerosis in the setting of T1DM in both humans and mice. This is particularly important because ASOs to APOC3 are under investigation in humans, raising the possibility that it may be possible to reduce CVD risk in T1DM patients by lowering APOC3 levels. Our preliminary data strongly support the hypothesis that APOC3 accumulation in APOB100-containing lipoproteins, intermediate-density lipoprotein (IDL) and LDL, makes these particles atherogenic in T1DM. To test this hypothesis, and to lay the groundwork for a clinical trial of APOC3 ASO therapy in the prevention of CVD in T1DM patients, we propose two specific aims. First, we will determine whether levels of APOC3 in IDL and/or LDL predict incident CVD risk in the Pittsburgh Epidemiology of Diabetes Complications study, a large prospective study. Our proposed study is well powered with ~550 T1DM patients and >30% rate of incident CVD. These studies will take advantage of a state-of-the-art method we developedtermed calibrated ion mobility analysisthat quantifies molar concentrations of APOB100-containing lipoprotein particles in blood. Our primary analysis will be to determine if i) IDL-APOC3 and/or ii) LDL-APOC3 predict incident CVD. Second, we will perform detailed metabolic studies to determine how T1DM alters hepatic APOC3 production and VLDL turnover rates, and how this impacts the accumulation of APOC3 in LDL and IDL in humans with and without T1DM. Based on our mouse studies, we hypothesize that T1DM promotes increased levels of hepatic APOC3 production that impairs TG lipolysis, resulting in increased levels of IDL-APOC3 and/or LDL-APOC3. We will complement these analyses with comprehensive analyses of the metabolic factors (e.g., body fat composition, liver triglycerides, hepatic sinusoidal insulin concentration) that might regulate the concentration of APOC3 in LDL, IDL and VLDL. Identifying patients at increased risk for CVD should provide mechanistic insights into the pathogenesis of accelerated atherosclerosis in T1DM. Moreover, it would lay the groundwork for a clinical study of APOC3 lowering therapy in T1DM because it could target patients at high risk of CVD.