University Of California Los Angeles

NIH Research Projects · FY 2025 · 2022-08

Abstract: Single-photon (1P) epifluorescence miniaturized microscopy coupled with genetically encoded calcium sensors has allowed investigators to record the activity of large populations of identified neurons over days to weeks in freely behaving animals, answering fundamental questions in neuroscience. Our group's efforts with the UCLA Miniscope Project have allowed over 600 labs to build and use over 2500 open-source miniaturized microscopes with expanded capabilities at a small fraction of the cost of those offered by commercial versions, thus democratizing access. Yet, 1P miniscopes lack the lateral and axial resolution to image activity in fine structures such as dendrites and axons. In addition, 1P imaging is limited to superficial structures or requires removal of overlying tissue for imaging of deeper neurons. Two-photon (2P) microscopy has exquisite lateral and axial resolution and bypasses all of these obstacles. Recent advances in technology have made the construction of two-photon miniaturized microscopes for mice possible. However, the field of view (FOV) is still limited, and these microscopes require custom-built optics and cost several hundred thousand dollars to acquire commercially. We have designed and built a two-photon miniaturized microscope for mice, including a custom- made objective lens, that allows 2P imaging of an 800 micrometer FOV nearly quadrupling the FOV from the latest published 2P miniaturized microscope (Mini2P-V1). In this proposal, we will optimize this microscope and test it in freely behaving mice for axonal, dendritic and deep somatic imaging. This microscope will be tested in three labs. The Golshani Lab will test the scope with calcium imaging of thalamic axons in anterior cingulate cortex during social interaction. The Silva Lab will test the scope by performing dendritic calcium and glutamate imaging in retrosplenial cortex during memory linking. The Shtrahman Lab will test deep imaging capability by imaging dentate granule neurons through an intact CA1. We will also build a larger miniaturized microscope suitable for rats and non-human primates with expanded capabilities, including a higher numerical aperture (NA), large FOV and temporal multiplexing capability to allow volumetric imaging at high frame rates (MiniMux2P). This microscope will be tested by the Blair Lab to dissect the role of superficial and deep CA1 neurons of rats in navigation. It will also be tested in the Churchland Lab to image rat posterior parietal cortical neurons during decision-making tasks. Finally, we will disseminate the technology using our open-source wiki that has already disseminated miniscope technology to thousands of users. We will provide parts-lists, optical designs and methods for obtaining custom lens elements. As we have done before, we will educate users through online videos and hands-on workshops where imaging basics, surgical techniques and analysis tools are demonstrated. We hope these cutting edge, novel and open-source tools will allow investigators to extend their research beyond that of what is possible with currently available technology.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT A major challenge in neuroscience is to uncover how defined neural circuits in the brain encode, store, modify, and retrieve information. Adding to this challenge is the fact that neural function does not operate in isolation but rather within living, behaving animals. Great technological advances over the past decades have allowed researchers to begin to optically measure and modulate neural activity but these approaches are often limited to head-fix animals when studying neural function at spatial and temporal scales relevant to internal neural circuit dynamics. While a great deal of scientific and technological progress has been made, there is still much to learn concerning complex neural function, especially within the context of natural behavior. This knowledge gap, at least in part, is due to a lack of accessible tools for simultaneously modulating and observing large-scale neural circuits with single-cell precision in freely behaving animals. This project will fill this gap by developing open-source, head-mounted miniature microscopes with spatiotemporal illumination capabilities for both patterned photo-stimulation and improved neural imaging in freely behaving animals. We will develop a modular control and acquisition platform for native integration of neural and behavioral equipment to facilitate neural-behavioral experiments. Finally, this platform will be driven by a novel, automated, closed-loop processing framework for detecting, decoding, and manipulating neural and behavioral dynamics in real-time. The goal of this platform is to 1) significantly extend and improve upon current freely behaving neural imaging and modulation techniques and 2) provide an integrated framework for observing, controlling, and manipulating neural dynamics within the context of behavior. This approach has the potential to simultaneously “read” from and “write” into, potentially, any area of the brain, enabling fine-grained investigation of the causal role between neural activity and behavior. Our development will be guided by concurrent benchtop and in vivo testing at every stage of the development and optimization process. To maximize the impact of our efforts, all tools and technologies developed for this proposal will be open-source and shared widely with the scientific community through online resources and technical workshops.

NIH Research Projects · FY 2024 · 2022-08

Project Summary/Abstract Depression is a leading cause of psychosocial impairment, poor academic performance, increased risk of school dropout, and a greater likelihood of self-harm, suicide, and medical illness over the lifespan. Moreover, a striking 30% of college students experience clinical levels of depression, making this is a critical period to identify intervention targets to mitigate risk for this burdensome disorder. Although research has evaluated psychological and immunological mechanisms of depression, these factors are typically studied in isolation. Social Safety Theory is a compelling, integrated framework that hypothesizes that social stressors are uniquely predictive of increases in depression because social stress has a greater biological impact compared to other stressors. Further, this pathway can be amplified by negative social safety schemas characterized by interpretating social situations as conflictual, unreliable, and dangerous. This proposal seeks to test this integrated, multi-level model of depression etiology using the transition from high school to college as a quasi-experimental social stressor and an intensive longitudinal design. Briefly, self-report data collected daily and inflammatory data collected every three days over a 24-day period will be used to evaluate how trajectories of perceived stress, inflammatory proteins, and depression symptom change as a function of transitioning to college (Aim 1), test if negative social schemas predict individual differences in trajectories of perceived stress, inflammatory proteins, and depression symptoms (Aim 2), and investigate the extent to which changes in inflammatory proteins mediate the association between changes in stress and depression symptoms during this period and whether this indirect relation is moderated by negative social safety schemas (Aim 3). To test if these interrelations between stress, social safety schemas, and inflammation predict long-term depression outcomes, Aims 2 and 3 will be re-tested predicting depression diagnoses across the entire freshman year (Exploratory Aim). Participants will be 112 healthy, incoming UCLA freshmen will be recruited via e-mails from the Registrar’s Office. Starting seven days before moving onto campus, participants will complete daily self-report measures (stress and depression symptoms, n = 2,688) and blood draws every three days (n = 896). Trait social safety schemas will be measured on the first day and a diagnostic interview will be completed before the study and again at the end of the year. Consistent with the NIMH Strategic Objectives (SO), this multi-method study will define biological mechanisms of mental illness (SO 1); provide insight into how mental illness trajectories change during developmental transitions (SO 2); highlight three, multi-domain targets for prevention and intervention (SO 3); and contribute to the public health impact of NIMH by reducing rates of depression on college campuses (SO 4). A training plan has been designed that includes a breadth of experiences aimed to advance the applicant’s conceptual and technical expertise in social stress and related cognitive vulnerabilities (Training Goal 1), high frequency immunological data collection (Training Goal 2), and statistics suitable for intensive longitudinal data (Training Goal 3).

NIH Research Projects · FY 2026 · 2022-08

Abstract Congenital and acquired craniofacial defects are not uncommon. Demineralized bone matrix (DBM) has been widely used for the orthopedic repair. However, more extensive use of DBM is limited due to its particulate nature after demineralization and rapid particle dispersion following irrigation, resulting in unpredictable osteoinductivity. Viscous excipients are often employed to produce stable suspension of DBM particles, but such carriers are rapidly dissolved in a body and the localized effect of osteogenic components present in DBM such as bone morphogenetic proteins (BMPs) may not expect at the defect site. Although exogenous BMPs can be combined to enhance DBM capacity, its clinical application requires supraphysiological doses and has revealed significant adverse effects. Thus, there is a need to develop alternative strategies that can enhance the osteogenic potency of DBM. This study seeks to enhance bone regeneration capacity by incorporating DBM into a self-healing dynamic polymer network that combines physiological stability and pro-osteogenic properties. Upon BMP stimulation, BMP efficacy is greatly reduced due to the enhanced expression of natural BMP antagonists such as noggin. Thus, this study will further enhance the potency of BMPs present in DBM by abrogation of BMP antagonism through RNA interference for noggin. Cell-derived exosome mimetics (EM) will be applied as a bio- vector to deliver RNA interference molecules in a localized and efficient manner. The overall objective of this proposal is to devise a robust bone graft composite that can effectively repair bone defects by integrating DBM and noggin-silencing EM into polymeric carrier systems. To achieve this goal, we propose three aims. In Aim 1, we will develop a malleable and self-healing hydrogel based on the self-assembly of phytochemical-grafted chitosan with silica-rich nanoclays, where the decorated phytochemical drives dynamic intermolecular interactions for gelation and nanoclay works as physical crosslinker with osteoinductive property. By varying the ratio of phytochemical to nanoclay and the content of DBM particles, hydrogel/DBM composites will be designed and prepared by evaluating gelation kinetics, injectability and self-healing characteristics. The osteoinductive activity of the developed composite will be determined in vitro and in a rat calvarial defect. Next in Aim 2, we will harvest EM from MSCs transfected with noggin-directed siRNA and evaluate the synergistic effect of EM on DBM-induced bone formation. We will also conjugate EM to hydrogels via a click crosslinking reaction for more localized and prolonged noggin silencing effects. Finally in Aim 3, we will integrate DBM and EM loaded with noggin siRNA into self-healing hydrogels of phytochemical and nanoclay developed from Aim 1 and evaluate the ability of the bone graft composite to promote bone regeneration in more challenging environments using a mandibular defect model. Successful bone formation will be evaluated compared with commercial DBM products and recombinant BMP-2. Successful completion of these studies will identify a new strategy to improve clinical efficacy of current bone grafting by maximizing activity of BMP signaling in DBM-mediated bone regeneration.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Persons living with HIV infection (PLWH) have a 2-fold higher risk of myocardial infarction and are twice as likely to develop cardiovascular disease accounting for a significant global burden of disease. While the mechanism underlying this excess risk remains poorly understood, studies demonstrate that atherosclerosis in the setting of HIV is distinct and characterized by heightened arterial inflammation as assessed by FDG- PET/CT. HIV and antiretroviral medication can worsen cardiometabolic parameters. Thus a therapeutic strategy that can lower lipids, inflammation, and improve glycemic parameters may be even more advantageous in HIV. Bempedoic acid (BA, an inhibitor of ATP citrate lyase), is safely tolerated, significantly lowers LDL-C and inflammatory markers (on top of statin therapy), and is FDA approved for individuals with heterozygous familial hypercholesterolemia or with established ASCVD who require additional LDL-C lowering. Additionally, BA has a protective effect on glycemic parameters and may reduce adiposity. Given the key role of lipids and inflammation in atherosclerosis in HIV, the purpose of this proof-of-concept mechanistic trial is to evaluate the impact of BA on the biology of HIV-associated atherosclerosis. We will perform a randomized placebo controlled study of effectively treated PLWH aged 40 years and older with either known CVD or 1 CVD risk factor to study the effect of BA on arterial inflammation (assessed by FDG-PET/CT), lipid levels, biomarkers of inflammatory/immune activation, cardiometabolic indices, and non-calcified plaque in the coronary arteries (assessed by CCTA). This multicenter trial will include PLWH enrolled at UCSF, UCLA, and Univ. of Utah. Long term collaborators at MGH will serve as the core facility for the imaging end-points. We have 3 specific aims for the: Cholesterol and inflammation Lowering via BEmpedoic Acid, an ACL-inhibiting Regimen in HIV Trial (CLEAR HIV Trial): Aim 1: To determine whether BA can safely reduce arterial inflammation including carotid plaque as assessed by FDG-PET/CT; Aim 2: To determine whether BA improves cardiometabolic measures (lipid, inflammatory, glycemic and adipose parameters) among PLWH. Exploratory objectives will be to assess BA’s effect (vs. placebo) on glycemic as well as adipose tissue measures (HbA1c, HOMA IR, and adipose tissue volumes); Aim 3: To evaluate the impact of BA on non- calcified coronary plaque volume as measured by coronary CT angiography (CCTA) and to determine whether changes in arterial inflammation are correlated with reduction in coronary plaques. This application combines (1) a successful multidisciplinary team with a strong record of collaboration and expertise in studying interventions in HIV, (2) the ability to rapidly recruit subjects from existing HIV-infected cohorts, (3) leveraging of resources including study drug/placebo. Identifying novel therapies to reduce CV risk are essential to improve mortality among PLWH, and results from this study will form the groundwork for a future trial to evaluate the impact of additional reduction of LDL-C and inflammation using BA on clinical events in HIV.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Autism spectrum disorder (ASD) is characterized by disabling social impairments and restrictive, repetitive behaviors that emerge in early childhood and persist throughout the lifespan, affecting 2.2% of adults in the United States. As they age, autistic adults face a range of adverse outcomes, including significantly higher rates of chronic disease, neurodegenerative conditions, and early mortality. My recent electroencephalography (EEG) findings further reveal altered trajectories of functional brain aging in ASD, in line with reports of excessive cognitive aging. However, the mechanisms underlying these age-related declines remain unknown, and by the time that age-related decline manifests behaviorally and cognitively crucial opportunities for risk prevention have already passed. New ‘epigenetic clock’ techniques index the progression of cellular aging processes based on DNA methylation (DNAm) patterns, providing proxy measures of biological age that predict later cognitive, health, functional declines, and mortality. I will explore if these sensitive measures of individual aging trajectories may help to identify autistic individuals at high risk of poor outcomes before patterns of brain activity or behavior begin to change, specifically asking: (1) Is biological risk for poor aging outcomes increased in autistic adults at midlife? (2) Are variations in epigenetic risk linked to brain aging markers? (3) Which clinical and environmental differences during childhood and early adulthood contribute to biological risk variations in this population? The proposed career development award will allow me to address these aims through new integrated methods. Facilitated by a multidisciplinary team of expert mentors (Dr. Lord, Dr. Carroll, Dr. Geschwind, and Dr. Senturk), I will build upon my existing expertise in developmental neuroscience and ASD to acquire new training in epigenetic and longitudinal lifespan methodologies. I will collect epigenetic and neural (EEG) aging measures from a unique and deeply phenotyped cohort of individuals with (N=118) and without ASD (N=39) who have been prospectively followed since age two and are currently 32-36 years old (The ‘Early Diagnosis Study; EDX). Biological age will be quantified from saliva-derived DNAm patterns using three different well-established epigenetic clock algorithms. Brain aging will be measured using EEG markers of peak frequency (7-13Hz), which captures characteristic age-related oscillatory slowing. Together, these studies will inform potential strategies to identify and address age-related risks in ASD from earlier in development. The proposed training goals will be the catalyst for a novel and innovative research program focused on lifespan changes in ASD across multiple levels of measurement and lay the foundation for a longitudinal R01 investigation of epigenetic, neural, and cognitive aging in the EDX cohort. This research program will address a crucial gap in our understanding of long-term lifespan influences in ASD and provide crucial opportunities to mitigate long-term personal and public health burdens in the rapidly growing population of older autistic adults.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Advancing the field of head and neck surgery through research and education has long been the defining mission of this department. This training program seeks to extend that mission to fill the urgent need for surgeon- scientists in all areas of otolaryngology-head and neck surgery. Two residents per year will embark on an 18 month period of research skills development designed to lead to long-term success as academic surgeon- scientists. The training period promotes innovation, collaboration, and strong research ethics, all guided by close mentoring and an individual development plan personalized for each trainee. Additionally, a medical student training program will enhance the pipeline of diverse medical students with research interests who are qualified to match in Otolaryngology-Head and Neck Surgery, via a nine-month intensive research and mentoring program. We expect that this effort will increase diversity within our specialty as a whole, as well as locally. Finally, a “Communication Disorders” seminar series will disseminate state-of-the-art knowledge to trainees and faculty affiliated with this program and related departments. Research themes of the program are grounded by 20 successful faculty researchers and include: 1) Laryngeal Physiology and Voice Perception, 2) Hearing and Vestibular Function, 3) Head and Neck Cancer, 4) Bioengineering and Bioinformatics. Mentorship teams, comprising one full-time researcher and one physician- scientist, will collaborate to enhance the trainee's development in all aspects needed for an academic career. Milestones including coursework, grant-writing, presentations and research publications will ensure development of those skills needed to become successful and versatile researchers.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Apoptosis happens continuously along with the active clearance of apoptotic cells (ACs) by phagocytes or efferocytes, termed “efferocytosis”, to maintain tissue homeostasis. When efferocytosis becomes defective, uncleared ACs undergo post-apoptotic necrosis and release immunogenic epitopes and pro-inflammatory mediators, which leads to chronic inflammatory diseases. Our recent studies revealed a novel role of efferocytosis in accelerating tissue repair as it promotes inflammation resolution by inducing the biosynthesis of specialized pro-resolving mediators (SPMs) that stop inflammatory responses. Therefore, understanding how efferocytosis is successfully carried out is of paramount importance. Much has been learned about the mechanisms of AC recognition and uptake, but how efferocytes degrade ACs and process the metabolic cargo, e.g., cholesterol released from AC digestion, is incompletely understood. Moreover, although efferocytosis and endocytosis share common features, such as involving cytoskeleton rearrangement and intracellular transport of vesicular membrane-bound cargoes, whether efferocytes hijack the endocytic machinery to process AC- derived cargo remains uncertain. In our unpublished results, we found that resolvin D1 (RvD1), a docosahexaenoic acid (DHA)–derived SPM, enhanced the acidification of the AC-containing compartments (efferosomes) and LC3-II lipidation, key features in LC3-associated phagocytosis (LAP)-mediated corpse degradation. As our recent study showed that the activation of MerTK, the efferocytosis receptor, was required for RvD1 biosynthesis, these results indicate a novel role of MerTK-RvD1 signaling in LAP-mediated AC degradation. To study whether the key endocytic regulators—the C-terminal Eps15 Homology Domain (EHD) proteins comprising EHD1, EHD2, EHD3, and EHD4—are involved in efferocytosis-related events, we analyzed a single-cell RNA-sequencing (scRNA-seq) dataset from atherosclerotic lesions where a lot of cells undergo apoptosis and found that EHD proteins had heterogeneous expression with high expression of EHD1 and EHD4 in macrophages, the professional efferocytes. We further found that EHD1 enhanced the cell surface levels of the cholesterol efflux transport protein ABCA1 in macrophages during efferocytosis, which indicates that EHD1- mediated endocytic trafficking of ABCA1 may play a role in removing the excess free cholesterol released from digested ACs. Here, we propose to combine approaches in cell biology, biochemistry, mouse genetics, and functional genomics to determine the function and mechanisms of MerTK-RvD1 signaling in LAP and EHD proteins in efferocytosis-related events including maintaining cellular cholesterol homeostasis and controlling endocytic trafficking of MerTK. We will also perform unbiased genome-wide CRISPR screening to identify novel regulators of MerTK levels on the cell surface where macrophages receive ACs. Taken together, understanding these aspects of efferocytosis will shed light on key physiological and pathophysiological processes and suggest novel therapeutic strategies for diseases driven by defective efferocytosis.

NIH Research Projects · FY 2026 · 2022-08

Project Summary/Abstract Each year, colorectal cancer (CRC) is diagnosed in 147,950 Americans and is responsible for over 53,200 deaths in the United States (U.S). The foundation of CRC screening is the detection and removal of precancerous colon and rectal polyps, which reduces CRC incidence and mortality. One million Americans are diagnosed with high-risk neoplasia (HRN) during screening colonoscopy every year, a specific subgroup of colorectal polyps that are associated with a 2- to 5-fold increased risk for subsequent HRN, CRC, and death from CRC. HRN removal prevents CRCs and saves lives. Consequently, professional medical societies recommend that individuals with HRN undergo surveillance with repeat colonoscopy 3 years after HRN diagnosis. While many research efforts focus on increasing CRC screening for average-risk Americans, few studies address low surveillance rates in this high-risk group. Lack of surveillance after a HRN diagnosis is due to multiple factors, including patient (e.g., no knowledge surveillance is due), provider (e.g. task overload, interpretation of colonoscopy and pathology findings), and healthcare system (e.g., no tracking or recall of HRN patients at 3 years) barriers. Therefore, in order to increase HRN surveillance rates, we propose to implement and evaluate a multilevel, technology- assisted intervention that automatically and reliably identifies patients with HRN, prompts patients and providers when surveillance is due, and facilitates colonoscopy referral and scheduling. The intervention will be implemented in UCLA Health, a large academic integrated health delivery network with over 15,100 screening colonoscopies performed and approximately 1,810 HRN diagnosed annually. It harnesses the strengths of a multidisciplinary research team representing clinical medicine, health services research, medical informatics, natural language processing (NLP), population health, economics and implementation science. The specific aims of the proposed R01 are: 1) to gain stakeholder perspectives on our proposed multilevel intervention and assess potential barriers and facilitators to receipt of surveillance colonoscopy; and 2) to conduct a hybrid type 1 effectiveness-implementation cluster-randomized trial to assess the effectiveness, implementation, and cost of a multilevel intervention aimed to improve colonoscopy surveillance rates for patients with HRN. The proposed study fills an important gap in CRC prevention and focuses on a high-risk group that has been largely neglected in CRC research. Furthermore, this approach has the potential to change clinical practice, is easily portable for addressing other types of polyps and surveillance intervals, and can be adapted for other health systems that face the similar challenge of identifying and recalling patients at elevated risk for CRC.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Morbidity and mortality in COVID-19 is the result of an exaggerated inflammatory response causing severe tissue and organ damage. However, the biological mechanism for why some individuals progress from initially stable to ultimately critical condition has not been fully elucidated. This work proposes that the immune response at the site of infection during the earliest stage of infection plays a deterministic role on subsequent pathology; namely that a delayed or suppressed type 1 interferon response in the respiratory mucosa within the first few days of infection permits rapid viral proliferation with minor symptoms, but eventually leads to the high viral loads and exaggerated inflammatory response seen later in disease. Further, the proposed project will test whether a delayed interferon response is the result of previously documented age-related dysfunction. It also seeks to determine whether direct SARS-CoV-2 mediated disruption of host splicing can also suppress interferon signaling in the early stage of infection. To do this, the study will leverage a unique set of longitudinal paired nasal swab and saliva samples from individuals who are initially negative for SARS-CoV-2 infection, but become infected while being prospectively sampled with high frequency (twice per day) as part of an IRB-approved (#20- 1026) COVID-19 household transmission study that the applicant co-designed and co-leads (since August 2020) at the California Institute of Technology. The transcriptome present in these samples will be analyzed to measure gene and isoform expression from which leukocyte recruitment and activation, including through interferon signaling can be inferred, through the elusive early stage of infection and the full course of the illness. Longitudinal differential expression and differential splicing paths in patients as young as age 6, and of advanced age will be assessed to identify whether age-related differences in immune response (in particular, interferon signaling) lead to more rapid increases in viral load, and subsequent symptom severity. In addition, from an RNA sequencing library enriched for nascent pre-mRNAs, splicing defects such as intron retention will be measured, to identify whether virus-mediated disruption of splicing also leads to a suppressed or delayed early interferon response permissive of rapid viral proliferation. COVID-19 is a public health threat, and in line with the mission of the NIAID, the results of this study can provide mechanistic insights into the pathophysiology of SARS-CoV-2 infection, to potentially identify novel or more efficient targets for the prevention or treatment of severe disease. In addition, the proposed project offers an excellent training opportunity for the applicant to gain knowledge and skills in immunology, virology/coronavirus biology, mechanisms of human RNA splicing, and generation analysis and interpretation of RNA sequencing data, with mentorship from Dr. Rustem Ismagilov, Dr. Akiko Iwasaki, and Dr. Mitch Guttman, who are experts in the aforementioned fields.

NIH Research Projects · FY 2026 · 2022-08

Electroconvulsive therapy (ECT) is one of the most effective antidepressant non-invasive brain stimulation therapies for adults with major depression. However, a number of patients fail to respond despite adequate trials, and while clinically beneficial, ECT can produce adverse cognitive effects including amnesia, executive dysfunction, and verbal dysfluency. Previous single- and multi-site ECT-imaging investigations have been limited by insufficient sample size and/or non-standardization of methodology. Therefore, in answer to NIMH Strategic Objective 3.2 “Develop strategies for tailoring existing interventions to optimize outcomes,” our investigative teams have conducted clinical studies to develop standardized methods for acute ECT course administration, antidepressant and cognitive measures for phenotyping, optimal neuroimaging protocols and E-field modeling, and sophisticated analytic models to integrate and interpret the antidepressant-response and cognitive- impairment biomarkers. In this prospective study we propose the first investigation integrating multiple units of analysis including clinical and cognitive phenotyping, whole-brain neuroimaging, EEG, and E-field modeling to establish the mechanisms underlying ECT-induced antidepressant response (response biomarkers) and cognitive adverse effects (safety biomarkers), as well as to find the “sweet spot” of ECT dosing for optimal antidepressant benefit and cognitive safety. Adult patients with major depressive disorder (n = 230) will receive a standardized acute ECT course, complete clinical and cognitive measures and undergo structural and functional MRI at three time points (baseline, after ECT #6, and following treatment completion) and one-month naturalistic follow-up. All MRI data will be processed and harmonized identically at a central imaging core to ensure uniformity. We have three primary aims: 1) Determine the relationships between E-field strength, ictal power, and biomarkers; 2) Determine the relationships between E-field strength, biomarkers, and antidepressant outcomes; and 3) Determine the relationships between E-field strength, biomarkers, and cognitive outcomes. An exploratory aim will contrast antidepressant-response and cognitive-impairment biomarkers identified in the current proposal with magnetic seizure therapy and healthy comparison subjects. The overarching hypothesis of this investigation is that the E-field variability will explain antidepressant and cognitive outcomes. Public Health Significance: Successful completion of this project will verify the optimal ECT dose (the “sweet spot”) of 112 V/m within the right hippocampus which can then inform precision and individualization of ECT amplitude with “E-field informed ECT”. The standardized algorithms for E-field modeling can be generalized and widely disseminated. This proposal will result in a paradigm shift from “trial and error” approaches of ECT parameter selection to individualized, precision dosing to improve patient outcomes.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY/ABSTRACT Glioblastoma is one of the most lethal of human cancers, with very few long-term survivors and no definitive cures for this disease. Immunotherapy is an appealing strategy because of the potential ability for immune cells to traffic to and destroy infiltrating tumor cells. Although immunotherapies, such as checkpoint blockade, have revolutionized the treatment of several cancers, its benefit in GBM has been limited to small randomized trials in the neoadjuvant setting, and limited benefit in Phase III trials in the adjuvant setting. Recently, we published a surgical trial of neoadjuvant PD-1 antibody blockade, to address the immunologic effects of this agent in recurrent glioblastoma. While it was a small, randomized clinical trial, the median overall survival of the neoadjuvant treatment cohort was 417 d (13.7 months) from registration date, while that of the adjuvant treatment cohort was 228 d (7.5 months; hazard ratio 0.39). Neoadjuvant PD-1 blockade was associated with upregulation of T cell– and interferon-γ-related gene expression, but downregulation of cell-cycle-related gene expression within the tumor, which was not seen in patients that received adjuvant therapy alone. These findings suggest that the neoadjuvant administration of PD-1 blockade enhances both the local and systemic antitumor immune response and may represent a more efficacious approach to the treatment of this uniformly lethal brain tumor than traditional treatment in the adjuvant setting. While provocative, the benefit was restricted to patients whose T cell infiltration and IFN-γ signature was elevated. Our hypothesis is that the combination of CTLA-4 and PD-1 blockade in the neoadjuvant setting will significantly increase the tumor-specific T cell infiltration, allowing for PD-1 blockade to be more effective. To test this hypothesis, we will utilize our unique access to samples and patient data from an ongoing investigator-initiated clinical trial with PD-1 +/- CTLA-4 antibody blockade in the neoadjuvant setting for patients with recurrent GBM. The Specific Aims of this project are: • Aim #1: To evaluate the immunogenicity, toxicity, and clinical benefit for the combination of neoadjuvant CTLA-4 + PD-1 antibody blockade in recurrent GBM patients. • Aim #2: To evaluate how PD-1 and CTLA-4 mAb blockade independently modify the tumor microenvironment and the expansion of tumor-specific T cells following neoadjuvant checkpoint blockade in recurrent GBM patients. • Aim #3: To develop non-invasive imaging biomarkers of response or adaptive resistance in recurrent GBM patients treated with combinations of neoadjuvant CTLA-4 +/- PD-1 antibody blockade. This project could potentially be transformative, as a better understanding of how neoadjuvant immunotherapy alters immune responses within the tumor could teach us important lessons about the critical requirements for productive anti-tumor responses in glioblastoma and how adaptive resistance occurs in this disease.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the U.S. and ranges from simple fatty liver (or non-alcoholic fatty liver, NAFL) to the progressive form, non-alcoholic steatohepatitis (NASH). About 20-30% of subjects with NAFL develop NASH, which is caused by hepatocyte injury, hepatic inflammation, and resultant hepatic fibrosis. NASH can lead to life-threatening conditions, but is difficult to diagnose at early stages. Liver biopsy is the current standard to diagnose NAFL/NASH, but biopsy is invasive, has associated morbidity, and is limited by sampling errors and inter-observer variability. Many patients present with later stage NASH, adversely impacting outcomes and healthcare costs, which are estimated at $32 billion annually in the U.S. Magnetic resonance imaging (MRI), including elastography (MRE), is a technology that can non-invasively quantify hepatic fat (MRI proton-density fat fraction), iron overload (MRI R2*), and fibrosis (MRE stiffness). However, current liver MRI is challenged by motion artifacts and incomplete signal models, which can compromise the accuracy and reproducibility of the quantitative parameters derived from them. In addition, early tissue changes associated with NASH are not adequately characterized using conventional MRI. The common requirements of breath-holding and long protocols also severely limit the adoption of liver MRI in the clinic. Furthermore, the present clinical interpretation of MRI has limited ability to distinguish NASH from NAFL. The research teams at the University of California Los Angeles, University of Arizona, and Siemens have been leading the development of motion-robust radial MRI to quantify hepatic PDFF and R2*, T2 and T1, perfusion, and stiffness. The Siemens team has also developed deep learning methods for medical image processing and disease detection and classification. In this bioengineering research partnership project, the multi-disciplinary research team will investigate four aims: (1) Develop a robust motion compensation framework for free-breathing multi-parametric quantitative radial liver MRI; (2) Accelerate quantitative liver MRI scans through combined acquisition and joint modeling of multiple parameters, data undersampling, and deep learning-based reconstruction and quantification; (3) Develop deep learning models to accurately classify NAFL versus NASH and measure the degree of fibrosis based on quantitative MRI; (4) Prospectively assess the new quantitative MRI and deep learning technologies for classifying NAFL versus NASH and measuring fibrosis in patients, with respect to liver biopsy. The new free- breathing quantitative MRI and deep learning technologies developed in this project will accurately classify NAFL versus NASH and measure fibrosis using data from the entire liver and thus help to avoid liver biopsy, allow monitoring of treatment responses, and accelerate the development and implementation of new therapies.

NIH Research Projects · FY 2026 · 2022-08

Academic tracking is a widely used practice that groups students into classes according to prior academic performance. In addition to potential long-term impacts on education attainment, academic tracking may directly affect adolescent social networks and substance use behaviors. By grouping students together with similarly performing peers, tracking may reinforce school disengagement and risky health behaviors like substance use, violence, and delinquency among lower-performing students. However, no known studies examine the health implications of academic tracking nor tested whether interventions to modify tracking positively impact health. Advancement via Individual Determination (AVID) is a successful college preparatory program that works in part by “de-tracking” students. AVID targets students in the academic middle (typically B- and C-average students) who are not typically placed in high-achieving academic tracks and places them in rigorous college-preparatory courses, while providing them with academic support to ensure their success. In our pilot study, a handful of students within 5 public high schools were randomized to AVID. We found the program led to connections with more pro-social peers and lower odds of substance use and delinquency. When applied school-wide, AVID trains schools to ensure all students have access to rigorous college-preparatory courses. However, there are no studies testing the health effects of AVID’s school-wide program. We propose a longitudinal study of adolescents attending schools with widespread AVID implementation and matched comparison (low or no AVID implementation) schools (matched on location and student characteristics) from communities in Southern California. Participants will be followed for 4 years to test whether exposure to AVID leads to a) lower rates of 30-day substance use (primary outcome--defined as any alcohol, tobacco, vaping, cannabis, prescription, or illicit drug use in the prior 30 days), and other substance use behaviors, violence and delinquency; b) increased enrollment in college-preparatory course taking and healthier social networks (measured by fewer peers engaged in substance use, more peers engaged in school, and more school-related adults); and c) whether associations between AVID and substance use are explained by increased college-preparatory course taking and healthier social networks. We will follow 9th-12th grade students at intervention and control schools for 4 years, collecting administrative education data and health behavior and social network survey data as they progress through high school and transition to college and/or the work force. This study will yield critical knowledge that can inform education and health policy regarding academic tracking and the use of de-tracking interventions like AVID. This topic is in keeping with NIDA’s mission to develop and disseminate research that significantly improves drug abuse and addiction prevention.

NIH Research Projects · FY 2026 · 2022-07

The high levels of saturated fat in Western diets contribute to the risk for obesity, diabetes and cardiovascular disease. Consumption of a fat-rich mean triggers a surge in circulating triglyceride TG levels as well as an inflammatory response that last several hours. As a result, we may spend a majority of our waking hours in a postprandial state. Non-fasting/postprandial TG levels are an independent predictor for cardiovascular disease. Studies in humans of multiple ethnic groups, as well as in the mouse, indicate that biological sex is a key determinant of postprandial hyperlipidemia, with males experiencing higher postprandial TG levels and inflammatory response. Knowledge gaps remain in our understanding of the physiological and molecular processes that differ between males and females to influence postprandial lipid handling and inflammation. Furthermore, the components of biological sex (which include ovarian and testicular hormones as well as genetic sex determinants, the XX and XY sex chromosomes) have not been systematically investigated with respect to postprandial metabolism. Our preliminary studies indicate that a lipid meal leads to substantially higher and more persistent circulating TG levels in males compared to females regardless of time of meal administration (i.e., during a typical fasting or feeding period) or presence of gut microbiota (the sex difference persists in gnotobiotic mice). Male mice also experience an enhanced postprandial inflammatory response, characterized by increased circulating monocyte number and inflammatory gene expression in bone marrow cells. Preliminary mechanistic studies indicate that rates of postprandial lipoprotein appearance in the circulation are similar in males and females, but that lipid composition and lipolysis differ. Our studies in the Four Core Genotypes mouse model reveal that the sex difference in postprandial hypertriglyceridemia is associated with gonadal sex (levels correlate with presence of ovaries vs. testes), whereas postprandial LPS levels are associated with sex chromosomes (presence of XX vs. XY). We propose to identify physiological and sex-related mechanisms that drive differences in postprandial lipid metabolism between males and females. Aim 1: Identify the metabolic processes and bioactive lipid components that promote male-biased postprandial hypertriglyceridemia and inflammation. We will perform in vivo and ex vivo studies in male and female mice to identify the mechanisms that lead to sex-biases in postprandial lipid composition, lipolysis, and inflammatory activation. Aim 2: Elucidate the control of postprandial lipid metabolism and inflammation by gonadal sex and chromosomal sex. Our studies in Four Core Genotypes mice revealed that distinct sex components influence postprandial lipid levels and inflammation. We will test the hypotheses that estradiol and/or testosterone regulate postprandial TG levels (using gonadectomized mice and hormone replacement), whereas the sex chromosome complement is a determinant of postprandial endotoxemia and inflammatory responses (using the XY* mouse model to compare animals with XX, XY, XO and XXY chromosome complements).

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY The medial prefrontal cortex (mPFC) makes essential contributions to learning, mood, and decision making, including evaluating and responding to threats. Compared to other circuits, mPFC circuits undergo prolonged maturation that is likely necessary in order to support complex behaviors. Yet how the development of prefrontal circuits enables the maturation of adaptive behaviors is poorly understood. Moreover, its extended development renders mPFC circuits highly vulnerable to disruption by environmental insults such as early life adversity (ELA). Even after adverse environmental conditions improve, ELA can produce lasting alterations in cognitive and emotional processing that put individuals at risk for many psychiatric disorders including anxiety disorders and depression. Although mPFC circuits are clear therapeutic targets for psychiatric disorders, substantial gaps in our understanding of typical mPFC circuit development and how this maturation enables emergence of complex cognitive and emotional behaviors prevent us from designing effective interventions. Our research will fill this knowledge gap by combining threat avoidance behavioral assays with projection-specific optogenetic manipulations, activity monitoring, and a model of ELA to test the hypothesis that precisely timed maturation of PL-BLA and PL-NAc circuits determines age-specific avoidance behaviors, and by altering the trajectory of circuit development, ELA leads to age-specific alterations in learned avoidance. In Aim 1, we will use optogenetics to test the hypothesis that the behavioral contribtions of specific mPFC circuits change over development. In Aim 2, we will use fiber photometry to expose circuit and region-specific activity patterns during learned avoidance across development. Finally, in Aim 3, we will apply the aforementioned approaches to a model of early life adversity to understand how chronic early stress impacts the trajectory of threat avoidance circuit development. This proposal directly addresses a pressing need to understand the complex relationships between early adversity, mPFC circuit development, and adaptive threat resposnes. Our research can inform individualized mental health treatments that exploit a variety of developmental opportunities, depending on the stage at which symptoms emerge.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT Temporal lobe epilepsy (TLE) is the most common form of focal epilepsy and is often refractory to medical management. In addition to seizures, patients often suffer from cognitive comorbidities, with memory impairment being the characteristic deficit seen in TLE. Despite the severe impact on patients’ quality of life, the mechanisms underlying poor memory are not understood and therapies that directly address it are limited. Previous studies in the lab have identified impaired place cell stability in the pilocarpine mouse model of TLE, which could underlie the spatial memory dysfunction in these animals. The place cell deficits do not emerge until 6-weeks after the epileptogenic insult (i.e. pilocarpine-induced status epilepticus, SE), suggesting that there are pathological processes triggered initially that take weeks to degrade hippocampal coding. This delay presents an opportunity for therapeutic intervention to prevent the degradation of hippocampal spatial representation, and is an important timepoint to examine for a comprehensive understanding of the cellular and molecular basis of memory dysfunction in TLE. The main hypothesis of this proposal is that circuit reorganization precedes the changes in network properties that cause place cell dysfunction and memory impairment, and that modulating circuit reorganization can improve place cell functioning and memory. Given the great cell type heterogeneity in the hippocampus, we performed single nuclei RNA sequencing on hippocampal tissue to determine cell type specific transcriptomic changes. We identified differentially expressed genes (DEGs) a different time points after pilocarpine-induced SE and focus follow-up studies on astrocytes given their emerging role in cognitive processing. To enrich for changes relevant to memory impairment, we compared the astrocytic DEGs at 3-weeks to a previously characterized Alzheimer’s disease (AD) model. AD, like TLE, is a disorder afflicted by memory impairment and has prominent hippocampal involvement. Within the overlapping genes, there is a functionally linked subset with roles in proteolysis: Cathepsins B, D, and L (Ctsb, Ctsd, Ctsl) and cystatin C (Cst3). These candidate genes have each been implicated in AD pathology in humans, although whether their role is protective or pathogenic is not fully determined. Additionally, while they were identified through analysis of astrocytic DEGs, they are broadly expressed across many cell types. In this proposal, we will use a CRISPR-mediated gene deletion strategy to selectively knockout each of these candidates from astrocytes, microglia, and interneurons. The effects of the knockouts on performance on a memory task will be determined in Aim 1. In Aim 2, the different knockouts will be evaluated in terms of their effects on hippocampal electrophysiology (sharp wave ripple occurrence, theta phase locking) and their effects on place cell function. In Aim 3, histological studies will be performed to evaluate the effects of the knockouts on a cellular level, looking at mossy fiber sprouting and synapse quantity. These experiments will elucidate the role of the candidate genes on memory processes and determine what the relative contributions are from each cell type.

NIH Research Projects · FY 2026 · 2022-07

UCLA LIFT-UP (Leveraging Institutional support For Talented, Upcoming Physicians and/or Scientists), proposes to develop a research and mentoring program that will contribute to strengthening the scientific workforce pursuing research careers aligned with the NIDDK mission. Our leadership team has a strong record of accomplishment in supporting post-doctoral fellows and pre-tenured faculty in obtaining external research funding and progressing to research independence. The proposed multi-PIs (Drs. Duru and Mangione) co-lead a successful P30 Research Centers from the National Institute of Aging, which has recruited, engaged, and supported 61 early-career researchers. We will leverage our expertise in research, clinical practices, policy, and program evaluation in diabetes care and prevention, along with our prior center experience, to identify and/or develop talented early researchers throughout Southern California, Hawaii, and Guam. UCLA LIFT-UP builds on the scientific strengths in the UCLA CTSI, UCLA Department of Medicine, and the UCLA Fielding School of Public Health. We propose three LIFT-UP Program Cores - an Administrative Core (AC), Research Mentorship Core (RMC), and Research Support Core (RSC). The RMC and the RSC will each be co-led by a basic scientist and a clinical outcomes/health services researcher, to enhance coordination of research mentorship and research support for investigators across the translational research spectrum. To accomplish these goals, the UCLA LIFT-UP overarching Specific Aims are to: 1. Provide the comprehensive guidance required to promote successful academic advancement of 14-17 early career researchers through mentorship and pilot research awards in the areas of obesity and diabetes across the translational research spectrum; 2. Engage early career researchers in cutting-edge laboratory-based science that advances the mechanistic understanding of molecular and physiologic processes related to obesity, insulin resistance, diabetes, and related conditions; 3. Enhance research infrastructure aimed at improving care and outcomes for individuals with obesity and/or diabetes through the design, implementation, and evaluation of collaborative interventions and policies with health system partners. The goal of LIFT-UP will be to mentor and develop highly skilled early career scientists whose passion and careers are devoted to preventing and treating obesity and diabetes, and working in disease areas aligned with the mission of the NIDDK. We will achieve this by actively identifying and recruiting the most talented candidates from Southern California, Hawaii, and Guam, and integrating them into the strongest, most comprehensive faculty development program available.

NIH Research Projects · FY 2025 · 2022-07

) Novel computational and statistical techniques are essential for enabling the translation of recent scientific dis- coveries across the widening spectrum of observations from both conventional information sources (e.g., elec- tronic health records) and emergent methods (e.g., genomics, mHealth, etc.). Indeed, biomedical informatics and data science are now foundational to both clinical care as well as in the advancement of scientific knowledge related to human health and disease. These methods are empowering more individually-tailored insights that can better guide healthcare delivery – the mainstay of precision health. Still, there remains an effective transla- tional gap that must be overcome, as today’s biomedical informaticians and data scientists need to better under- stand how to develop algorithms and tools for equitable precision medicine across the wide diversity spectrum of individuals. The UCLA Biomedical Data Science Training Program for Precision Health Equity is aimed directly at this pur- pose, fostering a new type of scientist trained at the intersection of contemporary computational approaches, biomedical informatics, public health, and precision medicine. Our trainees will be equipped with a technical depth that embraces these areas alongside an ability to translate such approaches to affect the transform of healthcare policy and practice with a goal for equitable medicine for all patients. To that end, this T15 brings together leading scientists and clinicians from across our institution and key areas to provide training in a com- prehensive, interdisciplinary manner that offers students a core curriculum in topics in clinical informatics, trans- lational bioinformatics, clinical research informatics, and public health informatics. It will afford trainees opportunities to see the pragmatic issues surrounding precision health and to learn how to address these barriers through innovative research and engagement. Didactic coursework and hands-on research experiences are shaped to reinforce technical and communication skills, team science, and a deep appreciation for the socio- technological concerns increasingly intertwined with precision health. As biomedical informatics and data science evolves, this T15 sees to an important area of growth that must be tackled to better serve the larger populous. Our program also makes a further commitment to diversity and equity through our broad inclusion efforts – a fundamental consideration if precision health is to ultimately be representative of everyone. Our trainees will be instilled with the critical ability to be forward-thinking, independent scientists who effectively contribute to trans- formation, working to improve every individual’s well-being through improve computational methods. Building on our faculty’s extensive experience in mentorship and establishment of groundbreaking scientific directions, this T15 is set to establish new scientific leaders who will drive needed change to enable precision health paradigms. Modified

NIH Research Projects · FY 2025 · 2022-07

The mission of the UCLA Cell and Molecular Biology (CMB) Training Program is to provide essential training for the next generation of PhDs in the developing fields of genomics, proteomics, systems biology, quantitative and structural biology, stem cell biology, and bioinformatics so that they are poised for successful careers in the biosciences. In addition, the CMB provides students with operational and professional skills training in a collaborative and supportive environment. The CMB Training Program brings together students and faculty from the life and physical sciences who share a common interest in cell and molecular biology but whose interactions might otherwise be limited. The 37 CMB Training Program faculty are drawn from departments that span the UCLA School of Medicine and the UCLA College. These faculty are passionate about using cutting-edge multidisciplinary approaches to tackle complex biological questions at the cellular and molecular level. The proximity of research buildings and university-supported state-of-the-art shared resources, along with the collaborative spirit that defines biomedical research at UCLA, facilitate cross-discipline interactions and foster interdisciplinary research studies. CMB trainees are drawn from four PhD graduate programs: Molecular Biology Interdepartmental Program; Neuroscience; Biochemistry, Molecular & Structural Biology; and Chemistry. Each year, the CMB Training Program will recruit 12 new students to a two-year program. Trainees must take four foundational courses that were designed specifically for training in modern cellular and molecular biology research. The objectives of these courses are to train students to develop technical, operational, and professional skills that will prepare them for biomedical careers including: a broad understanding of the cell and molecular biology field and the pressing questions and modern approaches used to address them; rigorous experimental design skills; statistical skills for data analysis; analytical and critical thinking skills; oral and written communication skills; and an understanding of how to conduct safe research studies. Through a CMB career development course, trainees are exposed to broad biomedical careers and acquire knowledge on how to prepare and apply for these professions. The CMB Training Program also provides mentoring and structured cohort-building and training activities, that engage and support students and promote collaborative studies among trainees. Together, the CMB Training Program’s student-centered interdisciplinary research training, active learning curriculum, career-development activities, mentoring, and community-building activities contribute to the successful training of students for a broad array of biomedical careers.

NIH Research Projects · FY 2025 · 2022-07

Project Summary: Atrial fibrillation (AF) is the most frequent cardiac arrhythmia, and it is a major risk factor for ischemic stroke and provokes morbidity and mortality along with a significant economic burden. Although AF has been studied in various animals, the embryonic zebrafish has been the genetically tractable and optically transparent model to investigate electromechanical coupling during cardiac development. Like in humans, the action potential and the consequent myocardial contraction are also key indicators of cardiac function in the zebrafish. By virtual of its transparency, optical mapping has been a primary means to investigate the interplay between cardiac action potential and myocardial contraction to study the mechanisms of AF. Dysregulation of electrical and mechanical coupling is a significant factor underlying the pathogenesis and perpetuation of AF. Optical mapping of electromechanical decoupling in zebrafish is nontrivial because it requires simultaneous recording of fast propagating voltage waves and myocardial contraction. Particularly in a beating heart, the rapid myocardial contraction can easily blur the image—the motion artifacts superimpose the wave patterns appearing in the optical maps and can prohibit further analysis of the imaging data. Pharmacological uncoupling has been widely used to suppress heart motion. However, this makes studying electromechanical coupling impossible. Alternatively, post-acquisition synchronization approach records a z-stack of movies, each covering at least one cardiac cycle. After the recording is completed, one 3D cardiac cycle can be reconstructed by synchronizing the movies in time. Nonetheless, this method is inapplicable to nonperiodic movements, such as irregular heartbeats with AF. Therefore, there is an unmet need to develop innovative optical techniques for high-speed 3D mapping of electromechanical coupling in a rapidly and irregularly beating AF heart. To solve this problem, we propose to develop a light-sheet light-field tomography (light-sheet LIFT) technique for kilohertz 3D imaging of electromechanical coupling in zebrafish hearts undergoing AF. Our method has only recently become possible due to two emerging technologies, light-field tomography (LIFT) and light-sheet microscopy, both of which we have extensive experience with. We will integrate LIFT with light-sheet microscopy and enable high-resolution 3D imaging with an unprecedented volumetric frame rate. The resultant system, light- sheet LIFT, will provide enough spatiotemporal resolution to fully depict the interplay between voltage waves, myocardial contraction, and intracardiac blood flow in a pitx2c zebrafish arrhythmia model. We expect our method will advance the understanding of AF's fundamental mechanism from the electrical activities at a single- cell level.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Non-alcoholic fatty liver disease (NAFLD) affects 25% of the US adult population and is associated with obesity, insulin resistance and cardiovascular disease. Furthermore, approximately 20% of NAFLD patients (13-16 million people) develop liver fibrosis, which is associated with a higher risk of hepatocellular carcinoma, cirrhosis and liver failure. Unfortunately, there are few effective treatments for NAFLD, aside from lifestyle modification and liver transplant. Excess neutral lipids are stored in lipid droplets (LDs)–dynamic organelles that quickly expand and shrink depending on the metabolic needs of the cell. Importantly, LD morphology and abundance are determined by the fusion or lipolysis of existing LDs. Recent studies have led to the discovery of a novel protein named BASIC, an endoplasmic reticulum-lipid droplet protein that promotes a multilocular phenotype and increases lipid utilization in brown adipocytes. BASIC is expressed in white and brown adipose tissue, and liver. In adipocytes, BASIC inhibits LD fusion proteins CIDEA and CIDEC, preventing expansion of LD size. CIDEB, the primary CIDE protein expressed in liver, has been shown to promote VLDL secretion, decrease triglyceride and cholesterol synthesis, and inhibit b-oxidation in hepatocytes. However, the physiologic or pathologic contexts in which the liver requires small, BASIC+ LDs for optimal function remain to be determined. Interestingly, global, but not adipose-specific deletion of BASIC decreases fat mass in chow- fed mice, pointing to adipose-independent effects on systemic energy balance. Preliminary data indicate that BASIC is a PPARa target gene whose expression is highly induced by both fasting and western diet feeding. Acute overexpression of BASIC decreases plasma lipids, suggesting this protein regulates hepatic lipid metabolism. The proposed research plan will elucidate the hepatic mechanism of action by characterizing how BASIC regulates LD biology and define molecular interaction partners in vitro (Aim 1a). Furthermore, the pathway(s) affected by hepatic BASIC expression (beta-oxidation, lipogenesis, lipoprotein secretion) will be determined (Aim 1b). In vivo studies using gain- and loss-of-function approaches in mice will characterize the role of BASIC in fasting and in response to high-fat diet feeding (Aim 2). Completion of the proposed aims will provide insight for the function of a novel PPARa target gene, which will contribute to the growing understanding of hepatic lipid droplet biology and may reveal novel opportunities for therapeutics in dyslipidemia and hepatic steatosis.

NIH Research Projects · FY 2025 · 2022-07

Abstract (30 line): The overarching goal of this study is to identify effective metabolic based diagnostic and therapeutic strategies to improve the overall survival of patients with Non-small cell lung cancer (NSCLC). We propose to investigate the positron emission tomography (PET) tracer, 18F-BnTP as a novel metabolic diagnostic and to develop metabolic based therapeutic strategies targeting oxidative mitochondrial metabolism in therapy resistant KRAS/LKB1 and EGFR mutant lung tumors. NSCLC will claim the lives of ~130,000 in the US in 2021. Lung tumors frequently possess a high mutational burden, often rendering single agent therapies targeting oncogenic driver mutations unsuccessful. Furthermore, metabolically active subsets of lung adenocarcinomas (LUADs) bearing mutations in KRAS and LKB1 or EGFR are frequently resistant to immunotherapy approaches. However, regardless of the initials benefits from checkpoint inhibitors or targeted therapies, the majority of patients will eventually develop resistance to therapy. We rationalize a different approach to overcoming therapy resistance in NSCLC – namely the classification of tumors by their metabolic signature. Here, tumors are grouped and targeted by their metabolic dependencies rather than solely by their genetic alterations. NSCLC is a metabolically heterogeneous disease and tumors utilize both glycolytic and oxidative mitochondrial metabolism to grow. The mitochondria are the site of cellular bioenergetics and oxidative phosphorylation (OXPHOS) and are essential for lung tumor initiation and maintenance. Due to a lack of in vivo imaging probes there is a gap in our knowledge at a physiological and mechanistic level of how mitochondrial bioenergetics are regulated in NSCLC. To address this gap, we functionally imaged mitochondrial activity in lung tumors utilizing the PET imaging tracer 18F-BnTP and demonstrate that it functions an in vivo biomarker of mitochondrial membrane potential (ΔΨ) and oxidative phosphorylation (OXPHOS) in lung tumors3. Importantly, by using 18F-BnTP PET imaging we are able to distinguish between OXPHOS dependent and independent lung tumors. Therapeutically, we have demonstrated that 18F-BnTP positive, OXPHOS-dependent LUADs are sensitive to mitochondrial complex I inhibitors. We hypothesize that 18F-BnTP PET imaging can be utilized to functionally profile mitochondrial bioenergetics and adaptive oxidative metabolism in therapy-resistant lung tumors to guide treatment with OXPHOS inhibitors. In aim 1 we will perform an in vivo dissection of mitochondrial bioenergetics in therapy-resistant LUADs. In aim 2 we will perform a structural and functional in vivo analysis of adaptive oxidative metabolism in therapy-resistant KRAS/LKB1 and EGFR mutant LUADs. In Aim 3 we will longitudinally profile oxidative metabolism in LUAD patients with advanced disease. The proposed work has relevance to human health in which we propose that 18F-BnTP PET imaging guided targeting and oxidative metabolism represents a new therapeutic strategy to overcome therapy resistance in patients with KRAS/LKB1 and EGFR mutant tumors.

NIH Research Projects · FY 2025 · 2022-07

Project Summary This project aims to better understand the regulation and function of RNA editing in cancer through analysis of existing omics data sets. RNA editing is a prevalent type of RNA modification where the RNA sequences are altered through insertion, deletion or substitution of nucleotides. In mammals, the most common type of RNA editing is adenosine to inosine (A-to-I) editing. Catalyzed by the ADAR enzymes, A-to-I editing is the most prevalent type of RNA editing in human, occurring in the majority of human transcripts. In the past few decades, great progress was made to understand the critical function of a small number of A-to-I editing sites in cancer-related genes, most of which alter protein-coding sequences. Owing to the recent advances in RNA-sequencing (RNA-seq) technologies and bioinformatic methodologies, an unprecedented number of A-to-I editing sites have been cataloged for various organisms. Importantly, widespread aberrant RNA editing has been reported in a number of cancer types. In addition, increasing evidence supports that ADAR and RNA editing levels are associated with patient survival or response to therapy. However, many questions remain, the most significant ones including the unclear mechanisms through which ADAR and RNA editing contribute to cancer-related pathways and the unknown regulatory mechanisms underlying aberrant RNA editing in cancer. In this project, we propose to extend our recent successes at developing and applying bioinformatic approaches in RNA editing studies to address the above challenges. We will capitalize on the large collection of RNA-seq data sets derived from different types of cancer samples. We will develop and apply novel methodologies to make full use of these data sets, complemented by further bioinformatic prediction and experimental validations, to predict and validate the molecular function of RNA editing and related regulatory mechanisms. This work will allow a previously unattained level of understanding of the molecular basis of RNA editing and provide new insights to the involvement of RNA editing in human cancer.

NIH Research Projects · FY 2025 · 2022-07

OVERALL - ABSTRACT The NIOSH Southern California Education and Research Center (SCERC) advances the field of Occupational Safety and Health (OSH) through a multi-campus, multi-disciplinary collaboration of eight programs across two University of California Campuses, UC Irvine (UCI) and UC Los Angeles (UCLA). The goal of the SCERC is to improve worker safety and health, productivity and workforce equity within Region IX and the nation. The Occupational Medicine Residency (OMR), Occupational and Environmental Health Nursing (OEHN), Industrial Hygiene (IH), and a proposed Occupational Epidemiology (OE) program offer graduate level academic and research training. The Targeted Research Training program provides specialized pre- and postdoctoral research training. A Pilot Project Research Training program engages and trains researchers new to the field from across Region IX. The Continuing Education program offers courses for practicing occupational health professionals and the Outreach program offers consultation and information on best practices and skills to the wider community of OSH professionals. The SCERC operates in concert with the UCLA and UCI Centers for Occupational and Environmental Health (COEH) which are state supported centers for research and teaching in occupational safety and health.