University Of California, San Francisco

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT The delicate lung alveolus relies on endogenous progenitor cells to maintain its function and regenerate after repeated insults. The alveolar epithelium contains alveolar type 2 cells (AT2s), which act as stem cells capable of self-renewal and differentiation into alveolar type 1 cells (AT1s), which cover the alveolus to facilitate gas exchange. AT2 to AT1 differentiation is a highly regulated process dictated by internal and external cues. Single- cell RNA sequencing has identified a “transitional AT2” on this differentiation trajectory, marked by Keratin 8 and senescence proteins like p16. In models of resolving lung injury, transitional AT2s gradually disappear as they differentiate into AT1s. However, in Idiopathic Pulmonary Fibrosis (IPF) or severe bleomycin (BLM)-induced pulmonary fibrosis, transitional AT2s persist and are thought to drive the disease. IPF is a devastating progressive age-related lung disease with no cure, and anti-fibrotic medications do not reverse fibrosis, suggesting another mechanism underlies IPF pathogenesis. Using a highly sensitive validated mouse p16 reporter developed in our laboratory, preliminary data show that p16 is rarely expressed in AT2s at steady state but increases in a subset of AT2s and their derivatives after BLM-induced lung injury. I will use a multi-transgenic mouse model of pulmonary fibrosis to determine the fate of p16-expressing AT2s in vivo and in vitro. Using a conditional knock-out of p16 in AT2s, I will test the functional role of p16 in AT2s during injury and repair. My hypothesis is that p16 has no effect on AT2s during homeostasis, but p16 is required for AT2s to enter a profibrotic state and during injury-triggered differentiation. Furthermore, I predict that AT2-specific loss of p16 will ameliorate the fibrotic phenotype in a BLM-induced mouse model. This research will demonstrate how manipulating senescence pathways affects AT2 stem cell function and could lead to new therapeutic targets to reverse pulmonary fibrosis. This proposal is part of a comprehensive training plan built with my mentors to develop the skills and knowledge needed to become a successful independent investigator in lung regeneration with an emphasis on chronic and age-related lung diseases. I have assembled a Career Development Committee composed of physician-scientists and basic scientists from pulmonology and aging who will serve as both scientific and career advisors. Upon completing this training plan, with support from my team and UCSF's strong research environment, I will be well-positioned to launch my career in regenerative medicine and contribute to understanding chronic lung diseases.

NIH Research Projects · FY 2025 · 2025-09

Unhealthy alcohol use plays an important role in perpetuating HIV transmission and exacerbating HIV-associated comorbidity. Prior research has often been hampered by the reliance on self-reported alcohol use, which can be biased and inconsistent. Alcohol biomarkers can provide objective and comparable alcohol measurements for behavioral and translational alcohol and HIV research. However, research is needed to determine how to use alcohol biomarkers as intervention components to provide objective feedback to motivate behavior change, and to determine alcohol biomarker cutoffs that predict worse health outcomes for people with HIV. Engaging persons most affected by alcohol use and HIV is key to understanding the potential impact of new research innovations on the lives of those with and at risk for HIV. There is also a need to train new researchers. We propose to establish the Center for Alcohol/HIV Innovations and Biomarker Research (CALIBER), a comprehensive program of research, training, and dissemination in partnership with those affected to (1) reduce the harmful effects of alcohol use on HIV prevention and HIV-associated comorbidities, (2) advance innovations in alcohol measurement, and (3) train the next generation of alcohol/HIV investigators. CALIBER will leverage expertise in alcohol/HIV biomarker, behavioral, and translational research with the extensive data, specimens, and infrastructure of the Multicenter AIDS Cohort Study (MACS) / Women's Interagency HIV Study (WIHS) Combined Cohort Study (MWCCS), which began testing for alcohol biomarkers in 2021. CALIBER will support three projects to address gaps in alcohol-associated HIV prevention and health comorbidity. Project 1 will develop and pilot test a motivational interviewing–based intervention that incorporates biomarker feedback to reduce alcohol use and increase uptake of HIV pre-exposure prophylaxis among couples at high risk for HIV. Project 2 will adapt and test a combined brief alcohol and expressive writing intervention to reduce traumatic stress and alcohol use among women living with HIV who are recruited from the MWCCS. Both projects will be conducted remotely (phone and online) to maximize scalability. Project 3 will examine the contribution of HIV and unhealthy alcohol use to steatotic liver disease in adults with and without HIV and the role of host genomics and gut microbiome, leveraging alcohol biomarker, historical self- report, and noninvasive liver imaging for ~3000 MWCCS participants. In addition, the CALIBER Pilot Core will train, mentor, and provide pilot funding for early-stage investigators, to conduct impactful alcohol/HIV research. The Dissemination and Community Engagement Core will translate research findings into sustainable solutions for individuals affected by HIV and unhealthy alcohol use. An Administrative Core will provide oversight and administrative, budgetary, resource, and scientific enrichment support to the other cores and projects and enable CALIBER to meet its goals.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract The long-term goal of this proposal is to modernize clinical methods for assessing fluid excess for removal during in-center hemodialysis treatment using newer assessment techniques. The experience of trial-and-error assessment of a “dry weight,” a weight at which a patient can tolerate fluid removal without the development of symptoms of hypovolemia or hypervolemia, a common practice for over 50 years, that may have a lasting effect on the ability to function for nearly 500,000 Americans who depend on hemodialysis for survival. Newer techniques for quantifying excess extracellular volume for removal during hemodialysis do exist. Small studies have suggested the potential utility of multi-frequency bioelectrical impedance spectroscopy (BIS) as a means to improve on clinical dry weight estimates to guide intradialytic ultrafiltration. Moreover, the rate at which fluid is removed during hemodialysis has been limited to not exceed 13ml//kg/hr based on observational studies on mortality. However, this association may differ relative to an individual's body size and may be more of a reflection of persistent volume overload rather than the just the rate of removal itself. The crude imprecise practice of dry weight assessment together with limitations in ultrafiltration rate may contribute to adverse patient experiences including persistent volume overload with peripheral edema and dyspnea on one hand, and intradialytic hypotension, fatigue, and muscle cramping on the other. Dialysis treatment day BIS estimation of excess extracellular fluid to direct ultrafiltration has the potential to improve patient-centered outcomes, the overarching premise of this proposal. We will gather evidence on the potential utility of BIS to improve the patient experience immediately following and during the post dialysis period. Our findings will inform future studies testing whether use of BIS to guide fluid removal during dialysis can improve how patients feel, function, and survive. We propose to compare dry weight estimation by BIS to usual care including estimate discordance on patient centered outcomes (Aim 1). We will also quantify the relative impact of high ultrafiltration rate and achievement (or non-achievement) of BIS-estimated target weight on patient physical function, sleep and quality of life (Aim 2) and examine clinical decision-making and patient perspectives on how UF management influences patients' dialysis experience using qualitative methods (Aim 3) among adults receiving hemodialysis in two metropolitan areas. The National Kidney Foundation's Kidney Disease Outcomes Quality Initiative (KDOQI) and the Kidney Disease Improving Global Outcomes (KDIGO) organizations are actively examining patient centered ways to improve care quality, the proposed work aligns with these efforts.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Brain arteriovenous malformations (bAVMs) are dysplastic vascular tangles that pose a significant risk for intracerebral hemorrhage (ICH). Current clinical and genetic prognostic factors remain imprecise in assessing hemorrhagic risk, underscoring the need for molecular classification. While bAVMs originate in endothelial cells, increasing evidence suggests that vascular stabilizing mural cells, such as smooth muscle cells and pericytes, are critical to bAVM stability and serve as cellular predictors of hemorrhage risk. Preliminary bulk RNA- sequencing findings from my laboratory show that mural cell depletion in adult bAVMs is associated with an aberrant immune response, whereas pediatric bAVMs exhibit unique angiogenic-like features, including Notch activation and TGFβ downregulation, suggesting an alternative mechanism of mural cell loss. This project aims to develop a multi-modal atlas of the normal and bAVM vasculature to identify non-immune, angiogenesis-centered disease mechanisms that contribute to mural cell depletion and ICH risk, providing a foundation for precision medicine strategies. The proposed research consists of two specific aims. First, an integrated cell and spatial transcriptomic atlas of the non-malformed human cortical vasculature will be created by harmonizing existing single-cell datasets and performing spatial transcriptomics on surgically resected tissues. This will provide a comprehensive reference for cerebrovascular studies and enable the identification of spatially distinct mural cell subtypes along the arteriovenous axis. Second, a multi-modal study of bAVMs across the lifespan will be conducted to elucidate age-related differences in disease mechanisms. Single-nuclei RNA/ATAC sequencing will be performed on a large cohort of pediatric and adult bAVMs to characterize gene regulatory networks (GRNs) associated with ICH risk. Additionally, an in vitro bAVM organoid model will be developed using induced pluripotent stem cells (iPSCs) carrying KRAS mutations to functionally validate the roles of Notch, TGFβ, and other identified GRNs in mural cell depletion. Targeted perturbation experiments using CRISPR activation/interference will assess the effects of these pathways on endothelial proliferation, mural cell recruitment, and vessel stability. The anticipated outcomes include a refined molecular classification of bAVMs, identification of novel biomarkers for hemorrhage risk stratification, and mechanistic insights into mural cell depletion that inform future therapeutic interventions. By leveraging integrative genomics and advanced organoid modeling, this research has the potential to transform the clinical management of bAVMs, advancing precision medicine approaches for cerebrovascular diseases. This work aligns with the broader goal of improving cerebrovascular health and reducing the global burden of stroke and ICH-related morbidity and mortality and informs precision medicine approaches to mitigate the risks of bAVM-associated hemorrhage.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Up to 25% of individuals in the United States have low serum vitamin B12 levels, leading to hematologic impairment (megaloblastic anemia) and/or neurologic deficits (loss of coordination, memory loss, psychosis). Comorbid B12 deficiency is associated with worse neurologic outcomes in patients with Alzheimer’s disease (AD) and Parkinson’s disease. Therefore, improvements in the diagnosis and treatment of B12 deficiency, either as a primary disease or a comorbid condition, have the potential to substantially improve brain health at a population level. I recently discovered an autoimmune cause of B12 deficiency restricted to the central nervous system (CNS), termed autoimmune B12 central deficiency (ABCD). Using unbiased antigen discovery technology, I identified autoantibodies targeting the transcobalamin receptor (CD320) which inhibit cellular uptake of B12, an essential cofactor for hematopoiesis and myelination. Anti-CD320 is highly predictive (96% specificity) of low B12 in the CSF despite normal B12 in the blood and remarkably prevalent (~10%) in patients with dementia. However, the mechanism by which anti-CD320 impairs B12 transport across the blood-brain barrier (BBB), its effect on neurologic function, and its comorbid contribution to neurodegeneration remain unknown. In Aim 1 of this proposal, I will elucidate the mechanism of action of anti-CD320 at atomic resolution. In Aim 2, I will determine the effects of anti-CD320 on myelination and neurologic function. Finally, in Aim 3, I will measure the prevalence and penetrance of anti-CD320 in AD. A mechanistic understanding of ABCD may lead to substantial improvements in brain health, ameliorate the burden of unexplained neurologic disease, and identify a modifiable contributor to cognitive dysfunction in patients with comorbid neurodegenerative disorders like AD. With the support of my mentors and advisors at the University of California San Francisco, this K08 proposal will allow me to fill training gaps and acquire new technical, communication, and management skills necessary for my transition to an independent investigator role at the interface of autoimmune neurology and neurodegeneration.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Pregnancy and the early postpartum period are times when women, especially women with HIV, are at highest risk for tuberculosis (TB). To reduce TB burden among all people with HIV, WHO recommends 1) intensified case finding (“ICF,” systematic TB screening followed by confirmatory TB testing for all who screen-positive) at every clinic visit and 2) immediate TB preventive therapy (TPT) for eligible individuals who screen-negative, in- cluding pregnant women with HIV (PWWH). Both ICF and TPT are particularly important during pregnancy to reduce risk of poor maternal and fetal outcomes. However, there are two main problems with the current approach to ICF and TPT for PWWH. First, symptom screening – the most widely used screening tool for TB – has unacceptably poor sensitivity among PWWH, missing up to 70% of all PWWH with TB, and performs worse with repeated rounds of screening. Second, the optimal timing of TPT is unclear because studies of PWWH receiving TPT have reported conflicting results, leading to inconsistent international guidelines on the use of TPT in pregnant women. New options for TB screening (CRP, C-reactive protein) and TPT (rifapentine-based short-course regimens) are now recommended by WHO and data among a general population of people with HIV suggest better accuracy and tolerability, respectively, relative to traditional tools. Definitive evidence is now urgently needed to determine whether CRP and/or short-course TPT can improve pregnancy outcomes at antenatal clinics in Africa, where TB remains a leading cause of non-obstetric maternal death. As such, we propose a highly efficient and innovative study using a sequential, multiple assignment, randomized trial (SMART) design to conduct a two-stage randomized trial to determine the optimal TB screening strategy and optimal timing of TPT initiation for PWWH. For the TB screening trial (Aim 1), we will randomize 1,500 PWWH presenting for routine antenatal care in Uganda to either CRP or symptom-based TB screening for the duration of their pregnancy and up to 6 months postpartum. The primary outcome for Aim 1 will be the proportion of all TB cases detected. For the TPT delivery trial (Aim 2), we will further randomize screen-negative PWWH enrolled in Aim 1 to either immediate (antenatal) TPT or deferred TPT. We will compare composite safety outcomes (primary outcome of spontaneous abortion, stillbirth, preterm birth, low birthweight, neonatal death) and composite effectiveness outcomes (co-primary outcome of maternal 2-year TB incidence and all-cause mortality) of immediate and deferred TPT. For Aim 3, we will use data collected from Aim 2 and IGRA results to construct a third TPT arm (targeted TPT) where immediate TPT is targeted to IGRA+ (high TB risk) PWWH and deferred for IGRA- (low TB risk) PWWH. We will compare safety and effectiveness outcomes of targeted TPT to the two untargeted (immediate and deferred) TPT strategies. This work will generate important data that will be critical to the future development of pregnancy-specific TB control activities, thus changing standard practice to improve the health of 1.3 million PWWH who give birth each year.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Our main goal is to examine the implementation, effectiveness, and cost-effectiveness of two implementation strategies for integrating a community health worker (CHW)-facilitated hypertension telehealth intervention into routine primary health centers operated by Kenya’s Ministry of Health (MoH). Hypertension, a major contributor to global morbidity and mortality and a key driver of cardiovascular diseases, remains inadequately treated in Kenya due to multi-level barriers at the health system, clinic, clinician, and patient levels. We previously showed that CHW-led telehealth interventions, combining CHW home visits and clinician assessment over the phone, can significantly improve control of hypertension by mitigating patient-level barriers like transportation costs to clinics. Despite the evidence supporting CHW telehealth, gaps in knowledge about effective implementation strategies at multiple system levels limit its broader application. This study will assess two approaches aimed at enhancing uptake, effectiveness, sustainability, and cost-effectiveness of CHW hypertension telehealth in MoH primary health centers. Guided by the Consolidated Framework for Implementation Research, we have identified two preliminary sets of implementation strategies: a) core (training, protocol development, MoH leadership engagement for clinic supervision, and medication supply chain improvements) and b) enhanced (core + plus implementation champions among clinicians and CHWs, and practice facilitation). The rationale for these approaches is based on the premise that while core strategies may suffice to address knowledge deficits and ensure effective implementation initially, additional external facilitation and empowering local champions might be necessary to reinforce training, increase motivation, iteratively adapt workflows, troubleshoot challenges, identify ongoing opportunities for improvement, and sustain implementation over time. In Aim 1, we will use Implementation Mapping with stakeholders from community to health system levels to adapt and refine these strategies for broader testing. We will attend to adaptation differences between HIV clinic (existing chronic care infrastructure and electronic medical record (EMR)) and general primary care clinic (current care is generally episodic, no EMR) settings. In Aim 2, we will conduct a type 2 hybrid implementation-effectiveness cluster randomized trial across 16 MoH primary health centers and surrounding communities to compare the impact of core and enhanced strategies on the implementation penetration of CHW telehealth and population-level hypertension control at 18 months. We will also evaluate sustainment during a 12-month maintenance phase following cessation of external support for implementation strategies. We will stratify all analyses by HIV status and sex to understand the role of these factors in implementation and effectiveness. Lastly, Aim 3 will measure cost and determine the cost-effectiveness of these strategies using an established individual-based hypertension model. This study will generate robust evidence for the most effective and cost-effective strategies for implementing CHW-based hypertension treatment, improving BP control and reducing CVD at a population level.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Atopic dermatitis often presents in early childhood and is characterized by a heterogeneous disease course: while some children improve by adolescence, many others continue to have persistent disease and develop comorbid allergic, neuropsychiatric, and metabolic conditions. Prognostic information is needed for patient counseling and to improve the efficiency of clinical trials evaluating whether new treatments could impact long- term outcomes. We propose a series of aims designed to predict which children with early-onset disease are more likely to develop persistent disease and comorbid conditions. First, using machine learning methods, we will develop prognostic models for persistent atopic dermatitis in late childhood using a standardized set of easy-to-measure early life predictors in diverse birth cohorts from Uganda, Ecuador, and the United Kingdom. We will compare model performance across cohorts and evaluate whether a single, generalized model for persistent atopic dermatitis could be useful in diverse settings. Second, we will create another set of models for each cohort adding more detailed predictors such as genetic risk scores, environmental variables and bio samples that vary across cohorts and may be more costly to measure. We will compare the performance of the models with an expanded set of predictors to the models that use a more limited set of standardized predictors within each cohort to better understand the nature of data needed for atopic dermatitis risk prediction across diverse settings. Finally, we will expand the prognostic models to additionally capture atopic dermatitis comorbidities, including other allergic diseases such as asthma and allergies; neuropsychiatric conditions such as anxiety, depression, and attention-deficit hyperactivity disorder; and metabolic disorders such as diabetes, dyslipidemia, obesity and hypertension. The results will offer useful prognostic information for patients, address a critical need for atopic dermatitis research among diverse populations, and enable targeted enrollment to improve the efficiency of clinical trials. This is particularly timely because the FDA has endorsed the use of tools designed to enroll patients most likely to demonstrate disease persistence, and there are many new systemic treatments under development for atopic dermatitis that are being tested for children as young as 6 months old.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Despite tremendous success in the reliable identification of more than 100 genes carrying large risk for autism, an actionable understanding of the underlying biology has been elusive. Gene ontology analyses have repeatedly emphasized an enrichment of genes encoding proteins involved in gene expression regulation and neurotransmission. These areas have thus become a central focus for the field. However, ontology-based analyses rely on incomplete knowledge of gene function and pleiotropy, and therefore, have the potential to miss key aspects of the underlying biology. Indeed, our recent functional in vivo work has begun to elucidate additional roles for proteins annotated as chromatin regulators in directly remodeling microtubules and the cytoskeleton. In fact, there exists a “tubulin code” of post-translational modifications analogous to the “histone code” and ample examples of canonical chromatin modifiers that directly modify tubulin to regulate microtubule dynamics. We have generated preliminary data for five such autism-associated chromatin regulators suggesting that they have dual functions, localizing to and functioning at microtubule-rich structures such as mitotic spindles, neuronal growth cones, and cilia in a wide variety of cell types, including human neurons. Thus, we have developed the bold hypothesis that autism-associated chromatin regulators have dual functions regulating both histones and tubulins. To test this hypothesis, we will deploy innovative experiments leveraging unique advantages of the Xenopus experimental toolkit, complemented by state-of-the-art proteomic profiling of tubulin post-translational modifications in human cells. In doing so, this work will illuminate how high-confidence ASD-associated chromatin regulators function at microtubules and the underlying post-translational modifications of tubulin that mediate these effects. This will set the stage for future work exploring how these fundamental effects on tubulin cascade to influence other neuronal processes like neuronal migration, axon outgrowth, dendrite formation, synaptogenesis, and synaptic transmission. This work has the potential to completely reframe the potentially relevant functions of these genes to autism biology, potentially bringing together the seemingly disparate enriched annotations since microtubule dysfunction impacts neurotransmission. I am uniquely positioned to lead this project, leveraging my experience in ASD genetics, proteomics, and innovative use of Xenopus to model ASD gene variants and identify core underlying biology.

NIH Research Projects · FY 2025 · 2025-09

OVERALL COMPONENT Project Summary The main objectives of this NEI Core Grant for Vision Research is to support the study of the visual system's structure, development, and function in both healthy and diseased states. The ultimate goal is to prevent, alleviate, or cure vision-related diseases, as well as to restore lost vision, by utilizing advanced techniques. This will be achieved through three resource and service modules: I. Ocular Structure and Function (OSF) module: This module will provide NEI R01 investigators with the tools necessary for ocular phenotyping, including morphological analysis, light and electron microscopy, ocular imaging, and functional vision testing. Additionally, it will offer prototyping services. II. Advanced Imaging and Analysis (AIA) module: The goal is to visualize cellular dynamics of ocular tissues with high spatiotemporal resolution. This module will utilize state-of-the-art imaging modalities, such as spinning disk, multi-photon, and confocal microscopes, along with resources for quantitative image analysis and creation of visual content. III. Vision Bioinformatics & Computation (VBC) module: This module focuses on advancing vision research through bioinformatics and programming for customized research needs. VBC will support the analysis of functional genomics datasets, including Next-generation sequence (NGS) and "omics" data, and will establish custom-built software resources for advanced data processing. VBC will provide computational resources and user-friendly software packages for researchers of all levels of experience.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY In high burden countries, many patients with tuberculosis (TB) are never diagnosed or treated with effective drug regimens, leading to ongoing transmission and increased mortality. A primary reason is that current TB diagnostics are inadequate with key issues including inadequate sensitivity, high costs, inability to be used at lower levels of the health system and/or failure to identify drug resistance. No single test is likely to address all these limitations and therefore the World Health Organization (WHO) has described optimal test characteristics for different use-cases in the form of target product profiles (TPPs). To advance novel solutions for high priority TPPs, identifying promising technologies and linking their developers to experienced clinical study sites to facilitate evaluation and performance feedback is essential. The overall goal of the Rapid Research in Diagnostics Development for TB Network: Episode 2 (R2D2 TB Network II) is to address the critical unmet need for better TB diagnostics in order to close the “diagnostic gap” and thereby improve patient and public health outcomes. To achieve this goal, the R2D2 TB Network II will solicit, review and prioritize the most relevant novel TB diagnostics across different phases of development and across different use cases for evaluation during the award period (Objective 1). We will leverage our experienced Technology Team, our partnership with a biotech incubator to bring in technology innovators not already working in the TB field and our partnership with key stakeholders with connections to TB diagnostics developers. Analytical as well as clinical studies to assess accuracy, usability, acceptability and feasibility of earlier phase tests will allow for iterative test optimization (Objective 2). These studies will be nested, where possible, within large-scale, multi-center assessments of design-locked diagnostics to facilitate WHO policy review (Objective 3). The clinical studies will rigorously follow WHO guidance for specific use-cases as well as general guidelines for high-quality diagnostic evaluations. For design-locked diagnostics, we will complement the clinical studies with assessments of incremental value (including when combined with other tests) through health economic and transmission modeling studies (Objective 4) to further support WHO- and country-level policy reviews. To accomplish these objectives, the R2D2 TB Network II brings together investigators with a broad range of relevant expertise related to TB diagnostic research and 10 experienced clinical study sites in 9 high-burden countries that provide access to relevant populations for evaluating TB diagnostics. The PIs overseeing the network have deep expertise in coordination of multi-center studies, a demonstrated history of working with a broad range of product developers, experience in the review and endorsement practices of the WHO and regulatory authorities, and more than a decade of collaborative leadership experience. By fostering and supporting a strong, collaborative network of investigators, product developers and stakeholders across diverse geographic sites, the R2D2 TB Network II will advance the next generation of TB diagnostics.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT In December 2023, the FDA approved the first genome editing therapy, which involves isolation of hematopoietic stem and progenitor cells (HSPCs) from a patient with sickle cell disease, ex vivo CRISPR editing, and re- transplantation into the bone marrow. This represented a landmark achievement for the fields of stem cell transplantation, genome editing, and precision medicine. However, the advent of CRISPR in the clinic has revealed new bottlenecks that limit translation of these potentially curative therapies. Current ex vivo HSC editing relies on chemotherapy- or radiation-based myeloablation to make space for genome-modified HSPCs in the bone marrow, which can lead to prolonged immunosuppression, negative impacts on fertility, and possible malignancy caused by genotoxicity. This represents a major safety concern for patients, however without myeloablation it would not be possible to achieve sufficient engraftment of genetically engineered cells to have a beneficial clinical effect. In addition, cost alone ($2.2M for the FDA-approved therapy) may prevent access for most patients, the vast majority of whom reside in developing nations. Direct in vivo editing of HSPCs emerges as an alternative to time- and cost-intensive ex vivo editing and would avoid myeloablation altogether. However, therapeutic in vivo editing is most effective in the liver, and achieving sufficient in vivo editing in HSPCs remains elusive. To address these challenges, we propose a novel approach: use genome engineering to enhance production of the desired cell type and effectively lower the therapeutic editing threshold. This solution is inspired by a rare condition called benign erythrocytosis, where otherwise healthy individuals have elevated levels of red blood cells and hemoglobin without increased cancer risk or reduced longevity. The proposed strategy aims to recapitulate the effect of naturally occurring mutations in the erythropoietin receptor (EPOR) using genome editing. Here, we will develop multiple strategies to pair the expression of a hypermorphic EPOR with sickle cell disease correction strategies in patient-derived HSPCs. These strategies include base editor-mediated multiplexing editing as well as Cas9/AAV-mediated gene insertion, which will be benchmarked against the FDA- approved therapy. We then will investigate the potential for this technology to mediate reduced or eliminated myeloablation in mouse models. Finally, we will adapt our most effective strategy for direct in vivo editing in HSPCs using a novel reporter mouse model. This approach could allow fewer edited HSPCs in the bone marrow to achieve a therapeutic effect, which could enable ex vivo editing to be curative using reduced-intensity myeloablation or enable direct in vivo editing of HSPCs, bypassing the need for cell isolation and re-transplantation. By drawing inspiration from nature and leveraging cutting-edge genome editing technology, this approach may improve the efficacy and safety of potentially curative treatments while paving the way for more accessible therapeutic options in the future.

NSF Awards · FY 2025 · 2025-09

This I-Corps project investigates the commercial potential of brain monitoring software for novice-level stroke recognition in anesthetized patients. Approximately 35,000 anesthetized patients each year in the United States suffer a stroke during a medical procedure. Because such patients are unable to manifest symptoms of stroke, the physician is not alerted, and the stroke is subsequently left untreated. Untreated stroke in this setting results in unexpected and premature death, physical and cognitive disability, and great hardship to these patients and their families. Additionally, the total cost of these strokes is about $2.8 billion per year. Brain electrical activity monitoring during anesthesia would enable real-time stroke detection, but it requires continuous high-level-expert interpretation. The need for expert interpretation prevents the large-scale use of brain electrical monitoring for stroke. Software to enable novice-level brain monitoring during anesthesia removes the need for an expert interpreter and expands the use of brain wave monitoring. With its use, the physician is alerted to the presence of stroke the moment it occurs. By having the stroke recognized immediately, the patient can receive emergency treatment to relieve the cause of stroke. By quickly reversing the stroke, the patients will be spared the severe consequences of stroke including potential emotional hardship, disability, and death. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of the technology. This solution is based on the development of the Correlate Of Injury to the Nervous System index calculated from electroencephalographic brain recordings. The index is a method of summarizing brain electrical activity from multiple sensors on the scalp into a single number that is calculated every 4 seconds. The number – which ranges from 0 to 100 – indicates whether there is an area of brain that has impaired electrical activity resulting from stroke. Like a pulse-oximeter (which is used to measure blood-oxygen content), index values between 90-100 are interpreted as normal, indicating a healthy brain without stroke. As the index decreases into values below 90, the clinician recognizes the potential presence of stroke. As values progressively decrease, the likelihood of stroke increases, such that a value below 70 almost certainly indicates the presence of life-threatening stroke. Additionally, the index generates a visualization which displays the area of brain affected by stroke. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Dementia prevalence and disease course differs by sex. Yet, despite being the leading cause of young onset dementia, very little is known about sex differences in frontotemporal dementia (FTD). Our accumulating data suggest females with FTD show patterns of “cognitive resilience” consistent with delayed symptom manifestation, despite higher neurodegenerative burden. We hypothesize that female-specific biology drives the unique clinical course of women suffering from FTD. Our overarching goal is to deeply characterize understudied female biology to unlock new insights into FTD pathophysiology, precision treatments, and novel dementia risk and resilience targets for all brains. To do so, our proposal will leverage two deeply phenotyped, longitudinally-followed local and international cohorts of genetic and sporadic forms of FTD and healthy controls (n=1095). We will be the first comprehensive evaluation of biological sex differences in FTD. Our Aims will: 1) Prospectively assay longitudinal sex hormone levels in plasma (e.g.,17𝛽-estradiol, testosterone, progesterone), 2) Identify sex-specific proteomic signatures of FTD from cerebrospinal fluid and plasma, 3) Collect a new female reproductive health history measure (e.g., age of menopause, pregnancies), and 4) Develop disease progression models that incorporate sex-specific factors to precisely prognosticate FTD trajectories in males and females. This proposal contributes to a high priority, underserved scientific area. For instance, among neuroimaging papers published since the 1990s, <0.5% consider factors specific to the female body. This knowledge gap creates treatment inequities as evidenced by the currently available therapy for Alzheimer’s disease (Lecanemab), which shows less than half the efficacy in females compared to males. Our proposal will therefore address a major health disparity, advance precision approaches to FTD before treatment inequities arise, and leverage overlooked female biology to discover new insights into dementia prevention targets relevant for all brains.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY This is a K23 award application for Dr. David Bayne, an Assistant Professor at UCSF. His career goal is to become a clinician-researcher focused on studying and reducing disparities in treatment outcomes for urinary stone disease (USD). This award will provide him with training and research experience to: (1) leverage electronic health record (EHR) data to streamline the identification of patients at risk for being lost to follow up (LTFU) after diagnosis of USD in the emergency department (ED); (2) elucidate stakeholder perspectives on root causes of, and potential solutions for, LTFU; and (3) evaluate acceptability and feasibility of early risk identification paired with patient navigator support as a pilot intervention to reduce LTFU for USD. Dr. Bayne has assembled an ideal team composed of co-primary mentors, Dr. Marshall Stoller and Dr. Charles Scales, experts in clinical trial design and implementation for USD; and co-mentors Dr. John Neuhaus, an expert in biostatistics and prediction model development, Dr. Sara Ackerman, an expert in application of qualitative research methods, and Dr. Lilia Cervantes, an expert in clinical trial design and implementation for patient navigation interventions. Although disparities along lines of socioeconomic status (SES) have been consistently described in the diagnosis and treatment of USD, interventions to mitigate these disparities are lacking. Dr. Bayne will build on findings from his prior work showing a consistent, independent association between delays in urologic care for USD and multiple EHR-derived predictors that correspond to low SES (e.g. insurance, demographic information, community level data). Predictive statistical models will be employed to streamline the identification of patients at risk for LTFU for USD upon placement of an outpatient referral to urological care in the ED by leveraging EHR-derived clinical and social data covariates (Aim 1). In addition, semi-structured interviews with patients and their providers will be conducted to elucidate themes contributing to LTFU for USD (Aim 2). A paired LTFU risk identification and patient navigation intervention will be developed and piloted to assess for acceptability and feasibility (Aim 3). This work will be the foundation of an R01 proposal for a randomized control trial to assess LTFU risk identification and tailored patient navigation as an intervention to reduce disparities in care delays in the treatment of USD. Through a focused program of mentored training and coursework, the candidate will gain new and necessary skills to deliver these research aims and advance in his career and professional development. These skills include: (1) prediction modeling with advanced biostatistics, (2) qualitative and mixed methods research, and (3) intervention development for clinical trials. These skills will enable Dr. Bayne’s transition to independence by uniquely positioning him to identify, study, and intervene upon factors contributing to disparities in USD care outcomes in low SES patients.

NIH Research Projects · FY 2025 · 2025-09

Abstract Hirschsprung Disease (HSCR) is a severe congenital anomaly within the gastrointestinal (GI) tract, resulting from the absence of intrinsic nerves in the distal bowel, a condition medically termed as aganglionosis. HSCR arises from mutations in numerous genes that lead to defects in the enteric nervous system (ENS)—an intricate network of neurons and glial cells that governs the GI motility and secretory functions. While surgical resection remains the primary treatment, it often fails to rectify persistent GI dysfunctions, underscoring the need for a comprehensive understanding of the disease pathogenesis beyond the current insights provided by animal models. Building upon established protocols for differentiating human pluripotent stem cells (hPSCs) into enteric neurons and glial cells, this research aims to systematically explore the impacts of known HSCR mutations on ENS development, from cell fate specification to migration and neuronal maturation. Employing single-cell transcriptomics, functional assays and transplantation experiments in mouse models, the study seeks to unravel mutation-specific effects on ENS function and identify therapeutic targets capable of ameliorating the cellular phenotypes associated with HSCR. The anticipated outcomes promise to drive the inception of broad-spectrum therapeutic strategies, enabling the development of personalized treatment modalities based on the unique genotypic and phenotypic presentations of HSCR, potentially improving quality of life and outcomes for affected individuals.

NIH Research Projects · FY 2025 · 2025-09

Abstract Silicosis is a chronic and potentially fatal fibrotic disease caused by inhalation of crystalline silica. Recent worldwide outbreaks of silicosis among workers who cut engineered stones for countertops underscore the necessity of understanding the cellular and molecular mechanisms underlying silicosis, for which there is no effective therapy. In silicosis lungs, activated fibroblasts generate granulomas called silicotic nodules and persistently cause fibrosis, which deteriorates lung function over time. However, the mechanisms underlying fibroblast activation and progressive fibrosis in silicosis have not been well studied. In previous work, we used lineage tracing and single-cell RNA-sequencing (scRNA-seq) to show that alveolar fibroblasts are the major origin of fibroblasts induced by bleomycin injury and in silicotic nodules in a murine model of silicosis. In preliminary work that serves as the basis for this proposal, we revealed that, in response to silica, alveolar fibroblasts differentiate into two unique populations that are not present in bleomycin-induced fibrosis or idiopathic pulmonary fibrosis. One population is localized inside silicotic nodules, expresses high levels of ECM genes, and is characterized by the up-regulation of ectopic bone regulatory pathways, which have recently been shown to be important in driving silicosis. The other population is restricted to the periphery of silicotic nodules and is characterized by hedgehog activation. This study takes advantage of our original mouse tools that allow us to target each unique fibroblast subset specifically and aims to elucidate the lineage trajectories of these fibroblast subsets and how they contribute to persistent fibrosis in silicosis. We hypothesize that pathologic fibroblasts originating from alveolar fibroblasts in silicotic nodules utilize ectopic bone regulatory pathways to cause a malicious feedback loop resulting in persistent fibrosis, while hedgehog-activated fibroblasts restrict the expansion of silicotic nodules. In Aim 1, we will use our original mouse tools to lineage- trace pathologic fibroblast subsets to elucidate the lineage trajectories and how pathologic fibroblasts in silicotic nodules are maintained for chronic fibrogenesis. In Aim 2, we will conditionally delete bone regulatory genes and the principal hedgehog receptor in alveolar fibroblasts to determine the roles of the pathways and fibroblast subsets we found to be unique to silicosis. In Aim 3, we will build a fibroblast-centric cell atlas of human silicosis with our specialized scRNA-seq strategy and high-throughput in situ hybridization to further assess the translational potential of our murine studies. These studies should provide insights into the cellular and molecular mechanisms driving silicosis, which could lead to novel therapeutic strategies for silicosis.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY The mammalian visual cortex is most often thought of, and studied, in the context of its image-forming ability, enabling detection and identiflcation of objects in the environment. Much less is known, however, about the cortical contribution to non-image forming visual pathways. A critical function of these pathways is to use visual feedback to guide motor actions. The pathology of non-image forming pathways is especially severe, as even the basic capability to maintain heading during locomotion depends on vision. The goal of this project is to elucidate the cortical neural mechanisms that stabilize locomotion using visual feedback. Locomotion and balance become even more dependent on vision during aging, as the vestibular system declines. As a result, locomotion becomes progressively restricted in the elderly population, leading to poor physical and mental health. Therefore, the signiflcance of understanding the cortical visual pathways that stabilize locomotion and their plastic capacity to compensate for sensory loss, is paramount. In the mentored phase of the award, I will determine the neural circuitry for using visual feedback to stabilize locomotion in mice. I have developed a novel paradigm to evoke course-corrective turns in freely locomoting mice using closed-loop manipulations of visual feedback. Using this paradigm I have found that the mouse primary visual cortex plays a key role in course-correcting locomotion, and I will test the hypothesis that it does so by acting on the superior colliculus through subthalamic feed-forward inhibition. Supported by my excellent mentor and collaborators, I will combine intersectional viral approaches, optogenetics and large-scale electrophysiology and imaging techniques to achieve this goal. In the independent phase of the award, I will address the capacity for the visual cortex to modulate the subthalamic circuits that stabilize locomotion to compensate for sensory loss. I will flrst test the hypothesis that real time changes in subthalamic activity, guided by visual cortex, can determine the effect of visual feedback on locomotion. Finally, I will perform vestibular lesions to determine how the visual cortex compensates by changing subthalamic dynamics. I am confldent that this work will shed new light on the ability of the visual cortex to provide Ʋexible control over innate behavior. The technical and scientiflc expertise that I will acquire during the training period of the award will be crucial for setting the basis of my independent research program. In addition to this intense career development training, the guidance from my mentoring team, as well as the collaboration and the rich intellectual interaction in the UCSF neuroscience community will ensure my successful transition into an independent investigator, focusing on the contribution of the mammalian cortex to innate and learned behavior.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY RNA modifications are altered in Alzheimer's Disease (AD) and related dementias. In particular, the post- transcriptional N6-methyladenosine modification of RNA (m6A-RNA) is dramatically dysregulated in human AD brains and in mouse and cell models of AD, and correlates with tau pathology. Tau physically interacts with m6A- RNA via the m6A-RNA binding protein hnRNPA2B1, and this interaction is increased in AD. However, we lack an understanding of the specific RNA sites that are differentially m6A-modified in AD and tauopathies, and of the downstream functional consequences and underlying mechanisms. In preliminary results, the Kampmann lab has uncovered new causal connections between tau, m6A-RNA, and hnRNPA2B1. In a genome-wide screen in human iPSC-derived neurons, knockdown of the m6A writers METTL3/METTL14 were among the strongest hits lowering tau aggregation. iPSC-derived neurons with the tauopathy-causing MAPT V337M mutation display increased m6A-RNA and altered levels and phosphorylation of many RBPs, including hnRNPA2B1. Intriguingly, we also observed mRNA mislocalization and misprocessing in MAPT V337M iPSC-derived neurons. Given that m6A and hnRNPA2B1 have been implicated in RNA trafficking and processing in neurons, we hypothesize that changes in m6A disrupt mRNA processing, trafficking and translation in AD/tauopathies, compromising neuronal function and survival. The Yeo lab has recently developed several innovative technologies that will enable us, for the first time, to simultaneously characterize RNA modifications and their downstream consequences in post- mortem brain tissue and model systems at scale and with unprecedented sensitivity. We will leverage our innovative methodologies to uncover m6A-RNA changes and their functional consequences in post-mortem brain tissue from subjects with AD and other tauopathies with unprecedented resolution, and to dissect the underlying causal mechanisms in human iPSC-derived neurons and mouse models of tauopathies. Thereby, it will advance our understanding of disease mechanisms and pinpoint potential novel therapeutic strategies.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Household waste burning, especially of plastics, is a major environmental and health hazard in low resource countries that lack a safe, reliable waste management infrastructure to dispose of waste. In rural Guatemala, 95% of households use solid fuels, primarily wood, for cooking and 88% burn waste as the primary means of disposal. Plastic waste is burned in cookstoves, often used as kindling, and is burned in outdoor fires as a means of waste disposal. Ecolectivos is a village-level cluster randomized trial using implementation research that is implementing 12-week community-level working groups in eight intervention villages with the goal of finding alternative strategies {e.g., refusing, repur1X>5ing, recycling, communi1y clean-ui:s) that reduce household burning cl plastic waste. In the biomonitoring RCT aim of our main study, we are enrolling 400 women of reproductive age {n=200 in eight intervention villages and n=200 in eight control villages) and measuring 24-hour personal exposures to air pollution {particulate matter {PM2.5) and black carbon {BC)) and urinary biomarkers {PAHs, phthalates, bisphenols) at baseline, 4-5 and 12-13 months. In the present study, we will deploy silicone wristbands, passive samplers that measure cumulative exposures to environmental chemicals, among 100 women who are participating in the Ecolectivos trial in rural Guatemala. Silicone wristbands will be worn for 8 days by 50 women in the two control villages and 50 women in two intervention villages at baseline and again at 4-5 months {after the 12-week working groups conclude in the intervention villages). In Aim 1, we will characterize cumulative exposures to compounds {polycyclic aromatic hydrocarbons {PAHs), phthalates, bisphenols, polychlorinated biphenyls, organophosphates, brominated flame retardants and PFAS Pre-cursors) extracted from the wristbands. In Aim 2 we will compare distributions of chemical compounds on the wristbands to filter-based PAHs and urinary biomarkers {phthalates and bisphenols) between women in the intervention group and the control group. The project is relevant to NIEHS's priority goal to address emerging environmental health issues. Plastic contamination is ubiquitous, yet personal exposures to burning plastic are as yet unknown.

NIH Research Projects · FY 2025 · 2025-09

BACKGROUND: There is an increasingly recognized role of stress in elevating disease risk. Differences in health may be in part due to greater exposure social stressors like early childhood stressors, and unsafe neighborhoods, through accelerating biological aging at the cellular level, although this has rarely been examining longitudinally. Our aim is to conduct a novel examination of stress and its links to rate of change in markers of cellular aging among women and their offspring during important transition periods (e.g., menopause, puberty) to understand potential pathways to early chronic disease development. Females and Blacks tend to have greater exposure to stressors. We have a remarkable opportunity to re-recruit a longitudinal cohort of Black and White girls who are now midlife women in the perimenopausal period. METHOD: This renewal proposal (R01AG059677) is to conduct a 40 year follow up of the prospective NHLBI Growth and Health Study (NGHS), a cohort of children to examine the life course and intergenerational transmission of cellular aging. Black and White girls were followed prospectively from roughly 10 to 20 years old, and we re-engaged them at 40 years old, with assessments of stress, cell aging and metabolic syndrome. We propose an assessment 10 years later, to predict change in biological aging from 40 to 50 years old. We will repeat assessments of cell aging (telomere length, epigenetic clocks, inflammation) and secondarily, metabolic syndrome in 624 NGHS women and cellular aging and BMI in their children. For Aim 1, we will assess which aspects of lifecourse stress—e.g., stressful life events, perceptions of stress, and neighborhood stress, are associated with rate of accelerated cellular aging of the women at age 50, testing differential associations of childhood, midlife, and cumulative stress. Secondary analyses will use causal mediation modeling to test potential mediating roles of early menopause, and health behaviors (nutrition, sleep, exercise). For Aim 2, we will assess the extent to which maternal stress and offspring stress are associated with change in offspring cell aging and BMI, and mediating roles of early puberty and health behaviors. SIGNIFICANCE & INNOVATION: This will one of the first prospective multigenerational studies to test lifespan stress predictors of distinct indices of biological aging and to assess stress effects on offspring epigenome. Given the prospective nature and range of stress measures, it has the potential to dramatically advance our understanding of lifespan stress on aging biology, and the roles of early puberty and menopause. A more granular understanding of the types and timing of social stress that impact biological aging processes is necessary for furthering our understanding of the malleable drivers of aging, and ultimately informing programs that promote health for all groups.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ ABSTRACT Tenofovir (TFV)-lamivudine-dolutegravir (TLD) has been first line HIV therapy worldwide since 2018. TLD is also used as second line therapy more recently in those with a history of prior treatment experience. However, in 2024, there is evidence of increasing rates of dolutegravir (DTG) resistance globally (4-8%), with rates up to 20% among those with virologic failure and treatment experience. Integrase strand transferase inhibitor (INSTI) resistance screening worldwide is therefore an urgent global priority, although HIV drug resistance (HIVDR) is still severely rationed in most low-and-middle-income countries due to cost. HIVDR is higher among people living with HIV (PWH) with virologic failure and laboratory-confirmed exposure to ART, demonstrating that high viral loads combined with ART exposure are more likely to signal HIV drug resistance (HIVDR) due to selective drug pressure from the antiretroviral(s) on the virus to evolve. The 2023 South Africa treatment guidelines now state HIVDR testing should only be requested after 2 elevated viral loads on TLD are documented and adherence is established using one of five objective adherence metrics; 1) a urine assay to detect tenofovir (TFV); 2) DTG levels in plasma; 3) tenofovir-diphosphate (TFV-DP) levels in dried blood spots (DBS); 4) clinic attendance; 4) pharmacy refill data. Our laboratory group at UCSF recently developed a point-of-care urine TFV assay that can assess with high accuracy whether TFV was recently ingested. However, the urine assay has not been rigorously studied for the prediction of resistance in PWH failing TLD to date. We thereby propose to test the ability of the urine TFV assay to triage HIV drug resistance testing in South Africa, comparing it to other metrics and assessing cost. We will first compare the prevalence of DTG resistance among PWH with prior treatment experience failing TLD with a positive or negative urine TFV assay, with participants recruited from large HIV treatment clinics in Cape Town and Johannesburg (Aim 1). We will then compare the ability of the urine TFV assay to predict DTG resistance with other adherence measures (other drug level assays, pharmacy refills, clinic visits) listed in the South Africa treatment guidelines, hypothesizing that the harder-to-collect metrics (pharmacy refills/clinic visits) will be less predictive of DTG resistance than the three more precise drug level assays (Aim 2). We will then examine the feasibility and cost-effectiveness of the urine assay in triaging HIVDR testing, since a scalable, cost-effective tool to determine those at the highest risk of HIVDR is urgently needed. The urine TFV assay is an exciting point-of-care diagnostic test which has already shown to improve HIV prevention and treatment outcomes in high burden HIV settings. If the low-cost urine TFV assay (<$2/test) is found to be highly predictive of HIV drug resistance in our study, the assay should be added to provincial, national, and international guidelines as a cost-effective, scalable tool to triage HIV drug resistance testing in this era of increasing concern regarding DTG resistance globally.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT Psoriasis affects 3% of the US population and a third of patients develop Psoriatic Arthritis (PsA), usually with a delay of 5-7 years. Delayed therapy leads to poorer PsA control, yet diagnosing PsA early remains challenging. Among the most effective therapies for Psoriasis and PsA are those that target IL23, IL17, and TNF; yet, these therapeutics incompletely control arthritis, indicating the existence of other arthritogenic immune pathways. We hypothesize that cutaneous inflammatory processes play a key role in triggering psoriatic joint inflammation, which evolves following cutaneous priming to include additional immune drivers. Addressing these hypotheses may reveal early biomarkers and novel therapeutic targets for PsA. To investigate how skin inflammation drives PsA, we developed a novel mouse model of immune dysregulation initiated within the epidermis. This model, termed A20eKO, uses temporal and keratinocyte- specific loss of the gene A20 (Tnfaip3), polymorphisms of which reduce its expression and are strongly linked to both Psoriasis and PsA in humans. A20 is a key negative regulator of innate immune signaling, and its loss unmasks spontaneous pathogenic tissue immune processes. Remarkably, A20eKO mice not only develop psoriasiform skin disease but also synchronous and fully penetrant PsA-like joint disease. We show that skin and joint pathology in A20eKO mice requires IL23, IL17A, and T cells, demonstrating that epidermal signals alone can drive arthritogenic T cells. This offers a clear model to study how psoriatic skin disease drives PsA. In Aim 1 we focus on the pathogenic T cells triggered by keratinocyte A20 deficiency. We will use single cell transcriptomics and paired TCR sequencing to determine the phenotype and clonal diversity of memory T cells that expand in A20eKO skin prior to clinical onset of dactylitis. Using bioinformatic methods, we aim to identify phenotypically similar cells in human psoriatic plaques, potentially representing skin-educated arthritogenic T cells. In Aim 2 we dissect the pathogenic contributions of two distinct signaling pathways, IL1/TLR and cGAS/Sting, that we discover as potently restricted by A20 in keratinocytes. Using A20eKO mice interbred with loxP-bearing alleles of MyD88 and Sting, we will determine how unrestrained keratinocyte signaling from these two pathways distinctly impacts psoriatic inflammation and PsA-like pathology in A20eKO mice. In Aim 3, we dissect how the immune mechanisms driving PsA-like inflammation evolve after triggering by cutaneous immune dysregulation. We find that delayed anti-IL23 treatment no longer prevents PsA-like inflammation in A20eKO mice. Using Spatial Transcriptomics we will determine the immune mechanisms that drive IL23-independent arthritis following cutaneous priming. Combining the A20eKO model with a novel method to reversibly trigger keratinocyte innate immune signaling, we will dissect how PsA-like inflammation responds once cutaneous immune triggers are reversed.

NIH Research Projects · FY 2025 · 2025-09

Project Summary The balance between regulatory and immunogenic immune responses is critical for liver homeostasis during perinatal life because dysregulation of either can lead to chronic viral infection from sustained immune suppression (as in chronic neonatal hepatitis B) or, to neonatal cirrhosis from unmitigated inflammation (as in biliary atresia). It is therefore essential to understand the mechanisms that maintain the delicate balance between regulatory immune responses and immunity in the perinatal liver as devastating sequelae of either too much immune suppression, in the case of chronic neonatal hepatitis B, or too much immunity, in the case of biliary atresia, leads to significant morbidity. In this proposal, we will investigate the role of the scavenger receptor MARCO in maintaining this crucial balance between regulation and immunity in the perinatal liver. Scavenger receptors are cell surface receptors found on phagocytic immune cells, and are important mediators of immune homeostasis, inflammation, and tolerance. We have identified an important role for the class A scavenger receptor MARCO expressed on macrophages in tuning immunoregulatory responses and inflammation in the perinatal liver. Our preliminary data indicates that MARCO is highly expressed on mouse and human liver macrophages, restricts anti-inflammatory myeloid cells, and worsens outcomes in a mouse model of perinatal liver inflammation. Our data also support the idea that MARCO restricts anti-inflammatory functions of macrophages by limiting responsiveness to the potent anti-inflammatory cytokine interleukin-10 (IL-10). We therefore hypothesize that MARCO restricts regulatory/anti-inflammatory immune responses by inhibiting responsiveness to IL-10. To address this hypothesis, we will use novel tools that involve technically challenging portal vein injections of a model antigen to quantify endogenous antigen-specific immune responses in neonatal liver and a CRISPR-Cas9 based method to ablate MARCO in primary human macrophages. Along with our established model of perinatal liver inflammation, we will use these tools to address the following Specific Aims: Aim 1: Determine whether MARCO restricts immunoregulatory responses by directly inhibiting the IL-10 receptor; Aim 2: Determine whether MARCO inhibits antigen-specific regulatory responses in the liver; Aim 3: Test whether MARCO limits resolution of perinatal liver inflammation by inhibiting anti-inflammatory monocytes. Completion of the proposed Aims will define the role of MARCO in perinatal hepatic immune function and will yield important insights into how macrophage-mediated immune signals maintain the balance between immune regulation and immunity in the developing liver. Understanding these signals is critical to elucidating the pathogenesis of highly morbid liver diseases in pediatric patients and is the next important step for being able to design potential therapies.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Hypertension (HTN) is among the strongest risk factors for cardiovascular disease and myocardial infarction, but only one-third of people with HIV (PWH) who receive care at a safety net clinic achieve hypertension control. People with severe financial challenges, particularly those with unmet subsistence needs like unstable housing and food insecurity, have higher rates of HTN and cardiovascular events in this setting. However, we still cannot pinpoint exactly how unmet subsistence needs influence where or why patients drop off the HTN care cascade. Here we propose a study among safety net HIV clinic patients. It uses an explanatory sequential design that includes quantitative survey data and qualitative contextual information. Our goal is to understand each stage of the HTN care cascade (diagnosis, treatment, and control) and overall cardiovascular disease risk among PWH who use a safety net clinic, emphasizing the unique HTN care needs of people with severe financial challenges. Results will be immediately usable by HIV safety net clinics; they will inform HIV/HTN integrated health care delivery models and the development of future HTN interventions.