University Of California Los Angeles

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Dobbs v. Jackson’s Women’s Health Organization, the U.S. Supreme Court’s 2022 reversal of Roe v. Wade, is the most consequential policy change for maternal and child health in a generation. At the time of submitting this grant, abortion had been banned or severely restricted in sixteen states, and efforts to impose additional bans and restrictions are in process in other states. Existing data limit more detailed analysis of who has abortions and the potential consequences of Dobbs for maternal and child health. The overarching goal of this exploratory proposal is to use a unique and powerful data resource to characterize the underreporting of abortion, develop new methods to improve estimates of abortion incidence, and facilitate more research on the consequences of Dobbs in national survey data. Our specific aims are to (1) document the individual-level determinants of abortion underreporting in surveys by comparing abortions measured in medical records to survey responses for the same individuals using a unique sample of study participants; (2) examine how abortion reporting in surveys changed after the Dobbs decision; and (3) explore the potential to improve national and subgroup estimates regarding the incidence of abortion in national surveys using evidence from aims (1) and (2). Achieving these aims will allow researchers to refine national survey estimates of abortion and understand the broader context in which these decisions occur. The proposed research will also inform a broader and deeper understanding of the disproportionate incidence of abortion bans or restrictions in the aftermath of Dobbs. The data underlying this proposal have already been collected and cleaned, making the proposed research highly cost effective.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT As of 2022, HIV infection and HIV-related diseases remain among the leading causes of death for children under five, despite the availability of antiretroviral therapy (ART). Children living in Sub-Saharan Africa face the highest mortality rate, with approximately 1 in 13 children dying before reaching their fifth birthday. Moreover, HIV- exposed uninfected (HEU) children (exposed in utero or during breastfeeding) have a higher mortality rate, more frequent hospitalizations, and more severe manifestations of diseases compared to those who are HIV- unexposed uninfected (HUU). It is estimated that there are 15.4 million HEU infants worldwide, with 90% of them living in sub-Saharan Africa. Cross-sectional age-stratified analyses have shown that the bacterial community composition in the saliva of HEU children changes from being similar to HIV-infected children under three years old to being similar to HUU children at age four and older. Furthermore, the incidence of oral diseases increases remarkably with age, which might be related to the development of oral microbiota. We hypothesize that the oral microbiome of HIV-exposed uninfected (HEU) children changes significantly during primary dentition (age 3-5 years) compared to the other two groups, children who are HIV-positive and in care (HIC) and those who are HIV-unexposed uninfected (HUU) during one year of follow-up. In this R03 study, we will collect oral swab samples from 360 children at three visits (1,080 samples in total). Duplicate samples will be collected and stored for future use. This proposed study focuses on the one-year change in the oral microbiome during the primary dentition stage among three cohorts in Kisumu County. Kisumu is one of the poorest regions in Western Kenya and has the third-highest prevalence of HIV/AIDS (19.3%) among children (0- 14 years of age) in Kenya. The research will leverage an important effort (parent study, R21TW012033) aimed at studying the dynamics of HIV progression and oral diseases. The proposed R03 study will provide the foundation for potential precise microbiota-targeted interventions to improve oral health-related outcomes among HEU children. Meanwhile, this study will strengthen the capacity and infrastructure for integrating both medical and dental care among HIV and HEU children in Kisumu, serving as an example of care for resource-limited areas. This study will also provide valuable preliminary data for an investigator-initiated R01 research project that aims to implement microbiome-based care into the routine care of HEU children. We will analyze the samples using 16S ribosomal RNA (16S rRNA) at the V4 region. This way, we can compare our results with samples collected in other regions using the same method. Additionally, we use 16S rRNA because it is the most cost-effective approach for a large number of samples and is easily scalable to resource-limited regions.

NIH Research Projects · FY 2025 · 2024-09

The goal of the proposed research is to develop, validate, and apply measures of state-level social context for Latino and non-heterosexual populations. Non-heterosexual Latinos have poorer HIV prevention and treatment outcomes than other populations. Moreover, they experience multiple forms of social rejection, which research by our team and others has shown to impact HIV-related outcomes. However, most work has focused on the individual level, while measuring and mitigating these phenomena at the state level is a key strategy for Ending the HIV Epidemic in the United States. Studies of social contexts impacting Latino populations are hampered by a lack of validated measures. Research on social contexts of non-heterosexual populations has expanded in recent years but the field lacks consensus on which domains, constructs, and items to include in analyses and on methods to ensure rigor and reproducibility of the research. Over this 4-year project, the investigative team led by MPIs Scheim and del Rio-Gonzalez will work with Scientific and Community Advisory Boards to build consensus on the domains and constructs needed to assess state-level social contexts affecting Latino and non-heterosexual populations through systematic reviews and a modified Delphi study (Aim 1); draw on the consensus developed in Aim 1 to develop and validate multidimensional indices of state-level social context for Latino and non-heterosexual populations (Aim 2); and examine relationships between social context and HIV prevention and treatment outcomes among non-heterosexual Latinos (Aim 3). This study will advance measurement science by using community-engaged, rigorous, and reproducible approaches to develop measures for Latino and/or non-heterosexual health research across multiple domains of health. We will make the validated indices and their underlying datasets publicly available for the research community, alongside consensus-based guidelines for selection and application of social context measures. A multipronged dissemination and research translation strategy will ensure the utility of study products and results for community organizations and researchers working to improve the health of Latino and/or non-heterosexual populations.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Chronic inflammation disrupts bone metabolism and promotes bone loss. Periodontitis and peri-implantitis are the most common inflammatory bone diseases in the oral cavity. In such an inflammatory environment, bone formation and bone resorption uncouple, leading to inflammatory bone damage, tooth loss, and dental implant failure. In this study, we propose a novel mechanism by which F-BOX Protein11 (FBXO11) regulates bone development and inflammation. FBXO11 is a protein-coding gene associated with otitis media. Additionally, it functions in a broad range of biological processes including melanocyte apoptosis, cell cycle regulation, cell migration, B-cell differentiation, and epithelial cancer progression. However, the effect of FBXO11 on bone development and inflammation has not been determined. Our preliminary studies in murine osteoblasts and genomic animal model showed that FBXO11 is a critical signaling molecule governing osteogenic differentiation by inhibiting Snail1/ lysine-specific demethylase 1 (LSD1). Furthermore, we found that the FBXO11/Snail1/LSD1 signaling axis is an important mechanism underlying inflammatory bone loss in cases of chronic inflammation, such as periodontitis and peri- implantitis. The novel mechanisms identified in our proposed studies will be critical for developing molecular strategies to prevent bone loss and promote bone regeneration in periodontal and peri-implant infection. Three specific aims will address the overarching hypothesis that FBXO11 regulates osteogenic differentiation in bone development and inflammation. Specific Aim1 will determine if FBXO11 is essential for osteogenic differentiation and bone growth by using FBXO11- overexpressing and knockdown osteogenic cells cultures, bone transplant experiment, and a conditional FBXO11 knockout mouse model. Specific Aim 2 will determine if FBXO11/Snail1/LSD1 regulatory axis contributes to inflammatory bone remodeling in the polymicrobial periodontitis and peri-implantitis animals. Specific Aim 3 will determine if we can render this FBXO11 axis as novel targets to treat periodontitis and peri-implantitis that represent significant health and economic burden world widely. We propose two innovative approaches, gene therapy by genetically engineering FBXO11 transgene in osteoblasts and a local intervention with a specific LSD1 inhibitor trans-2-phenylcyclopropylamine (2-PCPA), an FDA-approved antidepressant medication to prevent bone loss and promote bone regeneration. If this proposed study validates our hypothesis of repurposing of 2-PCPA to treat inflammatory bone disease, we will consider advancing the use of 2-PCPA to treat chronic periodontitis and peri-implantitis into clinical studies.

NIH Research Projects · FY 2025 · 2024-09

The leading cause of death in women in the United States is cardiovascular disease (CVD). Progress in reducing CVD in women has lagged behind progress in men. Obesity is a prevalent risk factor for CVD, and becomes more prevalent in women following menopause. We have shown that XX chromosome complement promotes increased adiposity in female compared to male mice, and this effect is amplified when gonadal hormone levels are diminished, as occurs after menopause in humans. We have identified the Kdm5c gene on the X chromosome as a key contributor to XX effects on adiposity. Kdm5c is expressed at higher levels in female compared to male humans and mice due to escape from X chromosome inactivation. Kdm5c encodes a histone demethylase that modulates chromatin structure and gene expression. Reducing Kdm5c gene dosage in XX females to that normally present in XY males reduces adipose tissue mass and alters histone marks in preadipocytes to favor induction of beige adipocytes and enhanced whole body energy expenditure. We hypothesize that sex differences in histone modifications throughout the genome, which likely change in hypogonadal states such as menopause, influence sex differences in adipose tissue function. We will investigate key questions regarding the role of Kdm5c gene dosage, and more generally, histone modifications and gene regulation, as determinants of sex differences in adipose tissue. Aim 1: Is Kdm5c gene dosage a determinant of weight/fat gain in hypogonadal states? We hypothesize that Kdm5c gene dosage is partially responsible for the impact of X chromosome dosage on weight gain when gonadal hormone levels are diminished. We will test the impact of Kdm5c gene dosage on weight gain, adiposity, and energy balance in female and male mice with preadipocyte Kdm5c deficiency under normal and reduced hormone levels. Aim 2: What is the effect of sex, gonadal hormones, and Kdm5c gene dosage on adipocyte histone and gene expression profiles at the tissue and single-cell levels? We will map the histone landscape and corresponding gene expression profile in adipose tissue to identify effects of sex, gonadal hormones, and Kdm5c gene dosage (A) at the tissue level using CUT&RUN and RNA-seq, and (B) and the single-nucleus level using Paired-Tag to simultaneously profile histone marks and gene expression in individual nuclei. Aim 3: How do sex and gonadal hormone status impact the histone landscape in human adipocytes? There is a dearth of information regarding the effect of sex on histone modifications in human adipose tissue, and whether changes in histone profile underlie alterations in adipose tissue following menopause/andropause. We will map histone profiles in adipose tissue of women vs. men, and in pre- vs. post-menopausal women. Our studies will shed light on how sex influences the epigenetic regulation of adipose tissue gene expression, composition, and function through histone modifications. This knowledge may inform therapeutic strategies to prevent excess adiposity and CVD risk in both women and men, including adipose tissue accrual in response to diminished hormone levels in mid-life.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Despite the tremendous success of the combination antiretroviral therapy (cART), the persistence of HIV reservoirs remains as the major obstacle to achieve HIV cure in infants. Current strategies mainly focus on the peripheral reservoirs; however, their efficacy is limited in the central nerve system (CNS) due to its unique anatomy. Extensive studies indicate that replication-competent reservoirs persist in the CNS under cART, causing neurological disorders in 15-55% of HIV+ individuals on cART. This issue is particularly concerning for infants, whose brains still grow and develop, leading to numerous neurodevelopment consequences and neurologic diseases. Therefore, there is an urgent need for effective treatment to suppress CNS infection in infants. One promising strategy is the use of broadly neutralizing antibodies (bNAb) directed to HIV-1, which have demonstrated potential in both viral suppression and reservoir clearance in animal models and clinical trials. However, the use of bNAbs for the CNS reservoirs is confounded by poor penetration of the blood brain barrier (BBB). Typically, antibody concentrations in the CNS are extremely low, with cerebrospinal fluid (CSF) levels at ~0.1% of those in blood. Our team has developed a novel nanotechnology, which we term nanocapsules, to encapsulate bNAb molecules within polymer shells, of which the surface contains abundant choline analogues. Such nanocapsules can effectively deliver bNAbs into the CNS by binding to the choline transporter on the BBB upon systemic administration, showing promising results in suppressing the virus in the CNS of non-human primate (NHP) infants. Our understanding of CNS infections and associated neuroinflammation during early infection and early treatment is still limited. Recent advances in molecular biology, nucleotide sequencing, and bioinformatics have enabled our team to develop methods to track infected cell clones with viral integration sites, revealing the impact of therapy on clonally expanded virus-infected cells. We are also exploring the use of a library of barcoded viruses for population-level analysis of infection in latent reservoirs and following HIV rebound. In this proposed study, we aim to evaluate the efficacy of bNAbs in nanocapsules against CNS infections and reservoirs, both with and without cART. The success of this study will shed light on the mechanisms underlying CNS infections in infants and demonstrate the effectiveness of bNAbs with improved CNS delivery in suppressing CNS and deep tissue infections and reservoirs. The findings could lead to interventions that can be initiated within the first few weeks of infection, a critical period associated with reduced mortality and slower HIV progression in infants.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Dr. Elinor Lee is a Pulmonary and Critical Care physician who is greatly interested in the role of lipid metabolism in surfactant homeostasis and pulmonary diseases. Dr. Lee is focused on mechanistic studies exploring the paradigm-shifting hypothesis that lipid dysregulation contributes to the pathogenesis of pulmonary alveolar proteinosis (PAP). Her long-term goal is to establish an independent research program as a physician scientist in the field of lipid biology and PAP and eventually, other pulmonary diseases. She is supported by her primary mentor, Dr. Tarling, a leader in lipid biology and inflammation, as well as her co-mentors and advisors who will provide their multidisciplinary expertise in training Dr. Lee in the theoretical and technical aspects of lipid metabolism and immunology. Through UCLA’s Clinical and Translational Science Institute (CTSI), Dr. Lee will have access to numerous career development seminars that address such topics as grant writing, manuscript preparation, and ethical research. She will also take graduate courses to obtain further training in gene editing, immunology, lipid metabolism, and biostatistics. Dr. Lee has the full support of her institution to carry out her research. This proposal outlines a 5-year research and career development plan that will prepare Dr. Lee to become an independent physician-scientist engaged in cutting-edge scientific research. This project aims to elucidate the mechanisms by which loss of granulocyte-macrophage colony-stimulating factor (GM-CSF) signaling impairs clearance of surfactant in alveolar macrophages. Dr. Lee’s prior work suggests that reduced expression of GM- CSF-dependent ABCG1, an intracellular cholesterol transporter that requires ATP to function, may drive reductions in cholesterol export by alveolar macrophages. Recent evidence, however, also suggests that reduced ATP production caused by impaired GM-CSF-dependent fatty acid catabolism may contribute to reduced phospholipid catabolism and/or cholesterol clearance. In Specific Aim 1, Dr. Lee will define the temporal sequence and kinetics of lipid abnormalities caused by loss of GM-CSF stimulation in vivo with a novel, inducible mouse model to identify the primary, pathogenic lipid abnormality and the secondary changes. Further, she will evaluate whether reducing cholesterol content in alveolar macrophages will restore their function to demonstrate that cholesterol accumulation due to impaired cholesterol clearance is the primary pathologic defect driving alveolar macrophage dysfunction seen in PAP. Aim 2 will focus on determining the mechanism of the primary and secondary lipid abnormalities caused by impaired GM-CSF signaling. Improved understanding of the relationship between loss of GM-CSF signaling and lipid dysregulation leading to surfactant accumulation will be crucial to the development of effective, innovative therapeutic strategies for patients with PAP and enhancing the fundamental knowledge of alveolar macrophages.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Vaping of aerosolized tobacco and cannabis products using electronic nicotine delivery devices (ENDS) is increasingly popular among adolescents and young adults. The associated emerging health crisis has significant systemic and oral health implications as nicotine (N) consumption is highly correlated with caries, the most prevalent oral disease in adolescents (57%) and young adults (82%), with cannabis use seemingly having a similar effect. Studies have shown that pure N enhances the growth and biofilm formation of the major cariogenic species Streptococcus mutans (Sm) and Candida albicans (Ca), which entertain a synergistic interkingdom relationship that is considered an important etiological factor in caries development. Limited recent studies on tetrahydrocannabinol (THC), the cannabis product favored by younger age groups, indicated that THC- containing cannabis smoke condensates stimulate the growth and biofilm formation of Ca. Despite emerging evidence supporting the pathogenicity-enhancing consequences of tobacco and cannabis product use, most research has used pure N instead of the more relevant vape aerosol (VA) containing N (N- VA) produced by ENDS. In addition, studies examining the effects of THC or VA containing THC (THC-VA) on Sm and Ca are very limited. Our published research and preliminary data show that the responses of Ca and Sm to pure N are distinct from responses to N-VA at the same N content (as determined by high-performance liquid chromatography). Therefore, we hypothesize that the enhancing effects of VAs containing N and THC result in different synergistic Sm–Ca cariogenic activities than the same concentrations of pure N and THC. In Aim 1, we will investigate the impact of N-VA and THC-VA on the Sm–Ca relationship compared with pure N, pure THC, and control VA (lacking N or THC) using well-characterized lab strains to evaluate co-adhesion, biofilm formation, extracellular polymeric substance production, acid production, and transcriptional responses. In Aim 2, we will validate selected findings from Aim 1 in clinically relevant Sm and Ca strains isolated from adolescent and young adult N and THC vapers as well as non-users with dental caries. We will characterize biofilm formation, acid production, and the highly differentially expressed genes identified in Aim 1. We expect to find that N-VA and THC-VA have distinct effects on the Sm–Ca relationship compared to their respective pure compounds or control VA. Evaluation of transcriptional responses in conjunction with cariogenic behaviors may provide physiological explanations for some of these differences. The results obtained from clinical strains isolated from different vaping environments will (1) validate findings obtained with Sm and Ca lab strains and (2) elucidate whether strains isolated from individuals who vape exhibit distinct responses to N-VA and THC-VA compared to strains isolated from non-users. The knowledge generated by this study will elucidate the effect of ENDS use on cariogenic activities and Sm–Ca interactions using clinically relevant parameters, contributing to meaningful public health communications regarding the impact of ENDS use on dental health.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Temporomandibular Disorders (TMD) represent a spectrum of painful disorders that afflicts approximately 5 to 10% of the US population with approximate annual healthcare costs of $4 billion. Degenerative disorders, internal derangements including disc displacement and hypo/hyper-mobility, and arthralgia of the temporomandibular joint (TMJ), which together are referred to as TMJ disorders (TMJDs), are common in many subjects with TMD. No definitive etio-pathologic diagnostics are currently available, and treatments are non-specific and largely palliative. While a subset of TMJDs occur in isolation, a substantial proportion are associated with comorbidities such as headaches and/or fibromyalgia. These two distinct patient cohorts together with the peculiar predilection of TMJDs for women of reproductive age as opposed to the late onset of similar disorders in other joints, point to unique and complex interactions of systemic and local factors. As such, multi-omic signatures and cell networks at the local and systemic levels will facilitate specific diagnostics and clinically meaningful stratification of patients between and within each of these cohorts. Currently there is a substantial void in connecting the molecular multi-omic signatures and cellular networks / interactions with clinical disease subtypes, etiopathogenesis, progression and severity. Our long-term goal is to identify local and systemic single cell and biofluid multi-omic molecular profiles from a spectrum of TMJD subtypes and severities to delineate relationships between clinical phenotypes with deep omic signatures and cell networks that will nucleate new directions for rational and precision therapies, prognostics and prevention. We expect that such multi-omic analyses combined with case control and longitudinal clinical and imaging data will provide critical insights on novel molecular signatures and networks associated with specific disease subtypes / severity and the interactions between cell subpopulations in perpetuating or mitigating disease progression. These goals will be achieved through two milestones-based phases involving (1) a UH2 feasibility phase to develop and implement protocols and quality assurance; recruitment of subjects and clinical disease categorization; sample collection to pilot sample handling and omic assays; data standardization, management and access; and developing a robust statistical plan; and (2) a UH3 implementation and discovery phase to identify distinct cell networks and molecular signatures towards a new rational classification of TMJDs and comorbidities; and to validate salivary omics biomarkers of these endotypes. Given the assembled expertise and plans, we expect that these studies will provide strong foundations and depth of knowledge needed to more rationally stratify patients into TMJD and comorbidity subtypes and severity categories; provide the basis for new directions of translational science in diagnostics, prognostics and therapeutics based on multi-omics of individual disease subtypes and the implementation of precision medicine for these patients; and define homeostatic and disease signatures that are specific to the TMJ as opposed to appendicular joints.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT We propose to develop new methods for the global identification of protein and RNP interactions with nascent unspliced RNA in human cells. We will use these methods to produce comprehensive binding site maps for several important molecules in splicing. In previous work, we isolated proteins and RNP’s from the chromatin fraction of cells to identify a new complex of splicing regulatory proteins, Rbfox/LASR, as well as a new U2 snRNP complex containing novel regulatory proteins. We further found that sequencing the pre-mRNA fragments bound within both these complexes allows the comprehensive mapping of their transcriptome-wide interactions. This “IPseq” mapping of U2/pre-mRNA interactions provides sensitive detection of intron branchpoints across the expressed transcriptome. We now propose to apply IPseq to develop binding maps of several key splicing components. We will continue ongoing studies of Rbfox/LASR and the U1 snRNP, and we will apply the method to the early spliceosome component: U2AF2. In preliminary data, we find that IPseq generates precise maps of the binding of both Rbfox1 and the LASR subunits on pre-mRNAs. Analyses of these data will yield important understanding of how the large Rbfox/LASR complex interacts with RNA, and how individual bound elements cooperate to generate a splicing regulatory code. The U1 snRNP recognizes the 5’ splice during early spliceosome assembly, binds within splicing silencer elements, and also acts to suppress premature cleavage/polyadenylation events in nascent RNA transcripts. Genomewide information on U1 snRNP binding is very limited, and the global analysis of its many RNA interactions will provide new insights into its multitude of functions. The U2AF heterodimer binds the 3’ splice site during early spliceosome assembly and is essential for the formation of an exon definition complex. A combined map of U1 snRNP and U2AF binding sites will allow their assessment as parts of possible cryptic exons and allow better prediction of mutational activation of cryptic splicing. By building a comprehensive map of the key factors for exon definition, our goal is to enable better prediction of pathogenic mutations affecting splicing. Finally, we believe that chromatin RNA IPseq can be broadly applied to many other cellular components.

NIH Research Projects · FY 2024 · 2024-09

This project will define the molecular spectrum of white matter vascular lesions that drive vascular cognitive impairment and dementia (VCID). Cerebral small vessel disease (CSVD) is a spectrum that begins in the vessel but progresses to include brain-wide changes in inflammation, dysregulated cell-cell communication, and cortical disconnection, ultimately leading to VCID. The goal of this research proposal is to develop a VCID Center With Out Walls (CWOW) that will advance a mechanistic understanding of the molecular pathways driving VCID while testing the efficacy of multiple VCID therapeutics. Converging evidence indicates that white matter vascular lesions resulting from CSVD modulate the location, severity, and progression of neuroinflammation, tauopathy, and neurodegeneration in AD-ADRD conditions. However, a key knowledge gap is the precise identification of molecular pathways both within cerebral small vessels and in brain networks at-large that mediate the cognitive impairment and dementia associated with white matter vascular lesions. Recent work and preliminary data from our group points to distinct molecular pathways driving inflammatory signaling and disrupted cellular communication in white matter endothelial cells, that produce disordered signaling in the microenvironment of cells around white matter vessels (the neurovascular niche), which leads to white matter axonal damage and then cortical disconnection acting through molecular pathways linking CSVD and AD. This vessel inflammation-to-cognitive impairment progression will be characterized using sophisticated viral, genetic, BRAIN initiative imaging technology, and human-to-rodent data alignment. Further, we will test three candidate therapeutics, developed by the PI’s, that interdict this vessel inflammation to brain disconnection progression. We will test this hypothesis using an unparalleled combination of prodromal, isolated, recurrent, and mixed ADRD rodent models of CSVD. We will use mechanistic studies aimed at identifying and validating molecular systems regulating white matter inflammation (SA1) as well as cortical dysfunction (SA2), and by developing a unique human single-cell sequencing dataset and applying translational VCID imaging biomarkers to rodent models of VCID to exploit regional differences in brain resilience (SA3). End point metrics will include: i) inflammatory vessel signaling cascades shared across prodromal CSVD models (aging, obesity, hypertension, and AD backgrounds); ii) inter-regional and inter-hemispheric measures of cortical disconnection that vary across lesion type and neurodegenerative comorbidity, iii) cellular, molecular, and imaging correlates of regional brain resilience to white matter vascular lesions, and iv) therapeutic efficacy of three novel VCID therapeutics targeting these pathways. This project and its research team leverages recognized leaders in VCID with relevant technical expertise to advance a VCID CWOW framework that will accelerate the identification of new therapeutic targets and diagnostic strategies to VCID.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Adverse childhood experiences (ACEs), traumatic experiences encountered during early life stages, can profoundly impact health and well-being into adulthood. ACE prevalence is high, with one in three children having at least one ACE. Children belonging to racial or ethnic minority groups are disproportionately affected by ACEs. Several health issues in children are strongly associated with ACEs, including asthma, sleep disturbances, infection risk, obesity, and behavioral issues. Additionally, children with multiple ACEs are at high risk of dental caries and untreated oral health care needs, even those with access to preventive dental care. The objective of this study is to investigate the possible biological factors underlying ACE-associated dental caries, with the overall goal of mitigating caries disparity in children with early-life adversity. ACEs function as biological stressors that affect neuroendocrine–immune (NEI) system responses. Recently, the brain–gut–microbiome axis, describing the bidirectional, biochemical interactions of the gut microbiome with the NEI system, was identified as a potential root cause of ACE-associated health conditions. Dental caries is a diet- and host factor–dependent disease in which a dysbiotic oral microbiome generates an acidic environment, leading to enamel demineralization. Our published work identified 10 salivary immunological markers that co- occur with caries-associated bacterial species and are significantly elevated in children with caries compared with caries-free children. Some of these markers are also elevated in children with ACEs. We propose the existence of a brain–oral microbiome axis, supported by multiple reports describing the salivary induction of NEI stress response markers that may alter the oral microbiome. We hypothesize that children with ACEs present with oral microbiome dysbiosis, increased dental caries, and immune dysregulation, including alterations in salivary stress or immunological molecules. This study aims to examine the associations among ACEs, the oral microbiome, dental caries, and salivary stress and immunological biomarkers in children. The innovative study design includes (1) strategies for enrolling a clinically meaningful, sex-balanced study population with an ACE prevalence that resembles the national average; (2) a comprehensive investigation of salivary markers, the oral microbiome, actual caries experiences, and covariate factors; and (3) a high-resolution metagenomics oral microbiome investigation. Study findings will contribute to understanding ACE-associated dental caries by identifying potential causative mechanisms at the oral microbiome level. The knowledge generated by this study has the potential to directly impact the clinical management of children with ACEs, promoting medical–dental integrated and trauma-informed approaches to ACE identification and caries prevention. Future policies may benefit from considering the contributions of childhood adversity to the oral microbiome and immune responses, facilitating the targeting of limited resources and relevant interventions for children with the greatest needs.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract: Early detection of Sjögren’s Disease (SjD) and its precursor autoimmune-Sicca Syndrome (AI- Sicca), are unmet clinical needs. Serum anti-Ro/SSA autoantibodies and biopsy focus score are two key classification criteria of SjD by the 2016 ACR-EULAR 1. SjD is a complex disease. Clinicians provide symptomatic treatment to patients as the symptoms of SjD are similar to other physiological and pathological changes in the body including menopause, drug side effects and fibromyalgia amongst others. This leads to a 3-year delay in initial diagnosis from the time the patients first seek medical care. In addition, there is currently no test for the diagnosis of AI-Sicca. In order to address this unmet clinical need, we have developed a saliva-based electrochemical assay, electric field-induced release and measurement (EFIRM), that can screen, earlier detect and risk assess onset of SjD, alternative to the current ELISA blood-based serology assay. Preliminary data demonstrated: 1) EFIRM can directly detect and quantify salivary anti-SSA/Ro52 autoantibodies in SjD patients; 2) Salivary and serum anti-SSA/Ro52 are correlated in SjD and AI- Sicca patients; 3) Salivary anti-Ro52/SSA IgA1 correlate to salivary gland focus scores. This R21 application will test the hypothesis in a large cohort of SjD and AI-Sicca patients that EFIRM, can: 1) detect anti-SSA/Ro52 and anti-SSB/La in saliva of SjD and AI-Sicca patients, and 2) salivary IgA1 to SSA/Ro52 is associated with salivary gland tissue destruction as measured by focus scores. Furthermore, monomeric versus polymeric forms for IgA1 to salivary SSA/Ro52 can discriminate SjD from AI-Sicca patients. Should this hypothesis be validated, the EFIRM salivary anti-SSA/Ro52 and anti-SSB/La assays can be explored for early detection of SjD and AI-Sicca in prospective multi-site clinical study as well as basic science mechanistic pursuits. In addition, saliva monitoring of IgA1 to SSA/Ro52 can be hypothesis tested for monitoring AI-Sicca progression to SjD in a non- invasive sample type (saliva). Three specific aims are proposed for this project. Specific Aim1: Acquire and Characterize Clinical cohorts of SjD, AI-Sicca and matched control subjects. Specific Aim 2: EFIRM quantitative assessment of anti-SSA/Ro52 and anti-SSB/La in saliva of SjD, AI-Sicca and Controls. EFIRM SSA/Ro52 and SSB/La autoantibody immunoassays will be performed on 210 SjD, 210 AI-Sicca and 315 controls subjects (blinded and randomized). Hypothesis testing of saliva SSA/Ro52 and SSB/La autoantibodies to discriminate 1) SjD vs AI-Sicca; 2) SjD vs Controls; 3) AI-Sicca vs Controls and 4) (SjD + AI-Sicca) vs Controls. Specific Aim 3: Quantification of monomeric and polymeric anti-SSA/Ro52 IgA1 in saliva of SjD and AI-Sicca patients.

NIH Research Projects · FY 2025 · 2024-09

Project Summary High doses of intensive rehabilitation therapy improve functional outcomes after stroke, but most patients do not receive this, for reasons that include limited access, difficulty traveling, and low motivation. Telehealth can address these obstacles. A recent StrokeNet trial found that a 6-week course of intensive home-based daily arm motor telerehabilitation significantly improved arm function as well as global function in patients averaging 4 months post-stroke, with efficacy comparable to dose-matched therapy delivered in-clinic. A definitive trial that compares telerehabilitation with usual and customary care is now needed. This issue will be addressed in the “Telerehabilitation In The Home After Stroke: A Randomized, Controlled, Assessor-Blind Clinical Trial (The TR-2 Trial),” a controlled, assessor-blind, randomized, phase III superiority trial that will recruit 202 patients with substantial arm motor deficits 4 months after stroke onset and randomize them to [1] a 6-week course of intensive daily arm motor rehabilitation therapy or [2] usual care. Aim 1 of the TR-2 Trial hypothesizes that adding a 6-week course of intensive arm motor telerehabilitation to usual care results in superior functional outcomes compared to usual care alone. The primary outcome measures arm function (Action Research Arm Test); the secondary outcome measures global function (modified Rankin Scale). Aim 2 will examine the predictive power of an imaging biomarker. Selecting the right patients is challenging in stroke clinical practice and trials due to the enormous heterogeneity of this disease. Clinical measures incompletely predict therapy gains, but studies from many labs have found that the extent of injury to the corticospinal tract predicts arm motor gains after stroke. The biological model underlying intensive arm motor telerehabilitation is that therapy activates multiple brain motor circuits, with the corticospinal tract being the final efferent pathway by which treatment gains are expressed, and so an intact corticospinal tract is needed to benefit from therapy. The specific hypothesis is that any benefit of telerehabilitation over usual care is a function of the extent to which the corticospinal tract is preserved. Aim 3 will evaluate the health economic impacts of the two treatment groups, with a focus on patient health-related quality of life, as the effects of telerehabilitation therapy must be considered in the broader context of healthcare utilization. Stroke remains a major cause of disability, and motor deficits are a major contributor. Rehabilitation therapy after stroke is generally provided at a very low dose, can be hard to access, and is often not very motivating. Our telerehabilitation program overcomes these barriers, was efficacious in phase I and phase II multisite trials, and will now be examined in comparison to usual care. The TR-2 trial is expected to generate definitive evidence that rehab therapy helps post-stroke at a time when many medical systems stop providing rehab care and so stands to change clinical practice worldwide.

NIH Research Projects · FY 2025 · 2024-09

HIV care outcomes and patterns of substance use vary significantly by geographic location. In regions like Los Angeles County (LAC), local, state, and federal policies have influenced access to healthcare and created issues, particularly for people living with HIV (PLWH) who also experience substance use (SU). Ensuring accessible, comprehensive, and patient congruent healthcare remains critical to improving both HIV and SU outcomes. Preliminary research, including our own studies, suggests that key factors influencing care utilization include the geographically dispersed locations of HIV and SU services, transportation access within large urban areas such as LAC, and limited alignment between patients and HIV care staff. While research shows that healthcare access is important, there are mixed responses of where PLWH prefer to receive care. These findings underscore the need for healthcare experiences that address geographic factors and are client responsive. Understanding how people navigate the geography of LAC to receive SU and HIV care and how peer case managers who know to navigate the geography and other factors may enhance access to care will provide novel insights into how PWLH and who use substances do or do not access treatment. We propose research and adaptation of an evidence-based intervention LINK-LA built on a previous NIDA-funded cohort, The mSTUDY (U01DA036267) and preliminary findings of an R21 on spatial connections conducted by this multi-disciplinary research team (DA049643). Applying mixed methods, we will interrogate the following (1) how public policies and practices have influence on spatial patterns and how such structural factors are associated with distinct patterns of movement across social and geographic space among PLWH in LAC and how these patterns influence where they choose to receive care. (2) How peer case managers who facilitate access to services and support to participants in the neighborhoods of their choice in LAC enhances use of services, and their HIV and SU outcomes by adapting previously proven interventions. Our proposed study will extend ongoing collaborations between the University of California, Los Angeles (UCLA), the University of California, Santa Barbara (UCSB), and a local HIV clinic resulting in a research program design that is directly informed by and responsive to patient needs. This collaborative team of scientists includes MPIs across all three career levels and local practitioners that represent an intentionally interdisciplinary and comprehensive response to improve use of and retention in HIV and SU treatment. Findings will be used to establish enhanced healthcare delivery models that improve HIV and SU outcomes.

NIH Research Projects · FY 2024 · 2024-09

Project Summary High doses of intensive rehabilitation therapy improve functional outcomes after stroke, but most patients do not receive this, for reasons that include limited access, difficulty traveling, and low motivation. Telehealth can address these obstacles. A recent StrokeNet trial found that a 6-week course of intensive home-based daily arm motor telerehabilitation significantly improved arm function as well as global function in patients averaging 4 months post-stroke, with efficacy comparable to dose-matched therapy delivered in-clinic. A definitive trial that compares telerehabilitation with usual and customary care is now needed. This issue will be addressed in the “Telerehabilitation In The Home After Stroke: A Randomized, Controlled, Assessor-Blind Clinical Trial (The TR-2 Trial),” a controlled, assessor-blind, randomized, phase III superiority trial that will recruit 202 patients with substantial arm motor deficits 4 months after stroke onset and randomize them to [1] a 6-week course of intensive daily arm motor rehabilitation therapy or [2] usual care. Aim 1 of the TR-2 Trial hypothesizes that adding a 6-week course of intensive arm motor telerehabilitation to usual care results in superior functional outcomes compared to usual care alone. The primary outcome measures arm function (Action Research Arm Test); the secondary outcome measures global function (modified Rankin Scale). Aim 2 will examine the predictive power of an imaging biomarker. Selecting the right patients is challenging in stroke clinical practice and trials due to the enormous heterogeneity of this disease. Clinical measures incompletely predict therapy gains, but studies from many labs have found that the extent of injury to the corticospinal tract predicts arm motor gains after stroke. The biological model underlying intensive arm motor telerehabilitation is that therapy activates multiple brain motor circuits, with the corticospinal tract being the final efferent pathway by which treatment gains are expressed, and so an intact corticospinal tract is needed to benefit from therapy. The specific hypothesis is that any benefit of telerehabilitation over usual care is a function of the extent to which the corticospinal tract is preserved. Aim 3 will evaluate the health economic impacts of the two treatment groups, with a focus on patient health-related quality of life, as the effects of telerehabilitation therapy must be considered in the broader context of healthcare utilization. Stroke remains a major cause of disability, and motor deficits are a major contributor. Rehabilitation therapy after stroke is generally provided at a very low dose, can be hard to access, and is often not very motivating. Our telerehabilitation program overcomes these barriers, was efficacious in phase I and phase II multisite trials, and will now be examined in comparison to usual care. The TR-2 trial is expected to generate definitive evidence that rehab therapy helps post-stroke at a time when many medical systems stop providing rehab care and so stands to change clinical practice worldwide.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY When making a decision we can use our understanding of available action-outcome relationships to prospectively evaluate the consequences of our potential actions and choose the one that is currently most beneficial. This goal- directed strategy is, thus, quite flexible, allowing us to readily adapt when circumstances change. For example, when ordering dinner, you consider how nutritious and delicious each potential meal will be and will avoid options that you’ve just had for lunch or do not meet your current nutritional needs. But we don’t always think about the consequences of our behavior. Usually this is fine. Such habits are a way for our brain to efficiently execute routine behaviors. But disrupted goal-directed learning and overreliance on habit can cause inadequate consideration of consequences, inflexibility, a lower threshold for compulsivity, and disrupted decision making. This can contribute to aspects of numerous diseases, including substance use disorder (SUD), obsessive-compulsive disorder, obesity, schizophrenia, depression, anxiety, and autism. Chronic stress can attenuate goal-directed learning and tip the balance of behavioral control towards habit. This is one major avenue through which chronic stress can predispose one to SUD and other mental illnesses. But we know very little of how chronic stress promotes habit formation. The overarching goal of this proposal is, thus, to expose the neuronal circuits through which chronic stress promotes premature habit formation. This work will help us to understand how chronic stress can promote maladaptive and pathological habits, which will be critical for understanding and treating SUD and mental illness. Filling this gap is especially important because chronic stress contributes to mental health disparities in the US. Recent evidence from our team has suggested that central (CeA) and basolateral (BLA) amygdala projections to the dorsomedial striatum (DMS) may be critical conduits for stress-potentiated habit. Based on prior and our preliminary data, our novel, comparative hypothesis is that BLADMS and CeADMS promote and oppose, respectively, goal-directed learning and associated neuronal activity in the DMS and that chronic stress amplifies CeADMS and attenuates BLADMS pathway activity to promote premature habit formation. We will test this with a multifaceted approach including projection-specific optical neuronal activity monitoring and manipulation, cell-type specific, cellular resolution, microendoscopic calcium imaging with pathway-specific chemogenetic manipulations, a model of chronic stress, and theory-driven behavioral tools. We will expose how ensembles of DMS neurons organize their activity during goal-directed and habit learning and how this is impacted by chronic stress. We will reveal the functions of two understudied amygdala-striatal pathways in goal-directed and habit learning and how they are influenced by chronic stress. More broadly, we will uncover neuronal circuitry through which chronic stress promotes habit formation. This will provide a critical basic science foundation for our long- term goal of understanding of how chronic stress is a conduit to pathological habits.

NIH Research Projects · FY 2025 · 2024-09

Project Abstract Diagnosis of Alzheimer’s disease (AD) is crucial for individuals to pursue treatments and plan for the future. Unfortunately, AD is underdiagnosed in community settings compared to the estimated prevalence from longitudinal cohort studies. AD underdiagnosis is exacerbated in understudied populations, including Hispanic/Latino (HL) and non-Hispanic African American (NH-AfAm) groups. Mining patients’ electronic health records (EHR) using machine learning may help identify patients with undiagnosed AD. Prior studies have identified individual comorbidities associated with AD. However, patients accumulate disease conditions sequentially accumulate over time. These pathways of disease accumulation are known as disease trajectories. Few studies have investigated disease trajectories in AD and none have comprehensively evaluated them in understudied populations. Disease trajectories derived from the EHR have the potential to predict undiagnosed AD as the next diagnosis follows from the prior diagnoses. Current models that predict AD do not utilize disease trajectories or account for AD underdiagnosis. EHR data can be complemented by genetics to determine AD risk. Incorporating polygenic risk with EHR data to identify undiagnosed AD has not been evaluated. To address these challenges, our objectives are to identify undiagnosed AD in non-Hispanic white (NH- white), HL, and NH-AfAm groups utilizing disease trajectories and genetics followed by validation and replication. We leverage existing resources from millions of patients with EHR and hundreds of thousands with genetic data linked to the EHR. We propose the following aims. Aim 1 will identify disease trajectories in AD. Aim 2 will identify undiagnosed AD based on disease trajectory while mitigating the bias of underdiagnosis in HL and NH-AfAm groups. We will validate patients predicted to have undiagnosed AD via multiple outcome measures and replicate our results in another EHR cohort, All of Us. Aim 3 will integrate polygenic risk with disease trajectories to detect undiagnosed AD. We will validate these patients predicted to have undiagnosed AD via multiple outcome measures, and replicate our results in All of Us. This proposal aligns with our long-term goal to realize precision medicine in dementias for all racial and ethnic groups by leveraging EHR, genomics, and computational tools. Our contributions will enable the application of disease trajectories in precision medicine, the detection of undiagnosed AD at UCLA and other health systems, and future work to reduce the bias of underdiagnosis in understudied populations.

NIH Research Projects · FY 2025 · 2024-09

Abstract Early poverty and lower education attainment are associated with increased rates of Alzheimer's Disease (AD), and lower cognitive functioning in later adulthood. Upwardly mobile youth who “strive” to rise above poverty typically have better cognitive outcomes in early adulthood. However, our preliminary data suggest that striving during adolescence can be stressful, particularly for females. Such stress may result in altered brain-gut axis signaling and systemic inflammation, which could set the stage for AD pathophysiology to begin unfolding before its onset in older adulthood. Women are at greater risk for striving-related stress, stress-related health problems, and critically, when they possess other AD risk factors such as apolipoprotein-ε4 (ApoE-ε4) gene status, they are more likely to develop AD. We hypothesize that due to sex-specific vulnerabilities (such as stress) females, but not males, with greater mobility opportunity will exhibit more physical health risks (namely obesity and inflammation as well as altered gut microbiome profiles) in early adulthood, which will in turn be negatively associated with cognitive outcomes and brain structure/function. We will test this hypothesis in a cohort of young adults (now age 23) who have participated in the Reducing Inequities through Social and Educational Change Follow-up (RISE-Up) Study since 2003. RISE-Up is a natural experiment using the admissions lottery of several high-performing public charter high schools to identify comparable groups of adolescents “randomized” into high- (mobility opportunity) and lower-performing (static) schools. The RISE-Up study provides a unique chance to better understand how adolescent mobility opportunities differentially influence adult cognitive health and AD risk in females and males. To this end, we will pursue three aims: (1) Identify the impact of mobility opportunity on young adult general cognitive ability (which we propose will link with later life resilience to AD), and test sex as a moderator; (2) Establish links between mobility opportunity, inflammatory markers, and microbiome functional potential, and determine whether those links are moderated by sex. Additionally, we will test whether sex and ApoE-ε4 status interact to predict inflammatory markers and microbiome functional potential; (3) Delineate associations between health (inflammatory markers, microbiome functional potential) and brain structure/function in early adulthood, probe their associations with general cognitive ability, and examine sex is a moderator.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The syndemic of alcohol use and HIV risk in pregnant and lactating people (PLP) threatens the health of mother, fetus, children and families in South Africa. PLP living with HIV who use alcohol may access antiretroviral therapy (ART) late or disengage with ART care, increasing the risk of vertical HIV transmission. PLP not living with HIV who use alcohol are at increased risk of HIV and may require targeted interventions to receive pre-exposure prophylaxis (PrEP) delivery and adherence counselling. Alcohol use also increases risk of HIV acquisition and poor ART adherence. The “mentor mother” (MM) intervention model is an evidence- based intervention (EBI) with demonstrated success in improving HIV and antenatal care outcomes. The model utilizes a task-shifting approach; positive deviant peers (mothers) deliver interventions to PBFW within and outside of the antenatal clinic. In the present study, we will collaborate with community partners to identify facilitators of alcohol use among PLP in SA. We will then adapt the MM model to create MM+ for PLP who use alcohol to evaluate the feasibility of an alcohol reduction intervention that includes HIV prevention and treatment (serostatus neutral) in a high alcohol using and HIV burdened community. Specific aims include: · Aim 1: Mixed methods assessment of perceptions of alcohol use in pregnancy and facilitators of alcohol reduction in pregnancy among multilevel stakeholders of PLP who use alcohol to identify clinic and community level facilitators that could be used to reduce the use of alcohol during pregnancy through in-depth interviews (IDIs) and brief surveys with drinking peers, intimate partners and focus group discussions with healthcare providers and community leaders. · Aim 2: Adapt and refine enhanced MM intervention (MM+) with potential intervention beneficiaries, providers and community leaders. Use community based participatory approach research principles to iteratively adapt, test and refine the MM+ intervention to integrate alcohol reduction content in one collaborative workshop session. · Aim 3: Evaluate the feasibility and acceptability of MM+ on reduction of alcohol use (primary outcome) and PrEP use (in PLP without HIV) and ART adherence (in PLP living with HIV) (secondary outcomes) in a pilot randomized control trial (RCT) in n=100 pregnant women who currently use alcohol. Primary outcome: Reduced alcohol use following the intervention (at 6m via phosphatidylethanol [PEth] levels). Secondary outcomes: PrEP and ART continued use at 6m via urine tenofovir levels at 6m. Secondary implementation outcomes: feasibility (provider, organizational and participant feedback), acceptability and fidelity to the intervention SOPs. Our study seeks to effectively reduce alcohol use and improve HIV care outcomes through community engagement and a novel participant informed intervention adaptation and delivery process.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY The goal of this proposal is to identify genes that affect behaviors associated with substance use disorders (SUD) using a novel mechanism-to-gene strategy. Standard genome wide association strategies applied to behavior in humans or model organisms have not so far provided much mechanistic insight into the origins of psychiatric disease. In this project we address this problem by genetic mapping of circuit-driven behavioral output. We examine a circuit involved in anxiety-related behaviors that are comorbid with SUD. Optogenetic activation of this circuit, from the ventral hippocampus (vCA1) to lateral hypothalamus (LH), results in large quantitative differences in anxiety-related behaviors in different inbred mouse strains. These differences are highly heritable and amenable to genetic mapping. Since a circuit's function arises from a limited set of cellular and anatomic features, identifying the genes that are involved in circuit function will generate hypotheses on the mechanisms underlying the behavioral differences and hence of SUD. We will identify the genes and likely mechanisms through three aims: 1) determine heritability and behavioral variation of optogenetic activation of the vCA1-LH pathway in eight inbred strains 2) genetically map optogenetically-evoked behaviors of the vCA1-LH pathway using 800 diversity outbred mice derived from those eight strains, and 3) apply quantitative complementation tests to find which genes act through the vCA1-LH pathway to alter the behavioral function of this pathway. These experiments will allow us to identify where and how in the brain genetic variation gives rise to behavioral variation, and will demonstrate the importance of integrating systems neuroscience and genetic approaches.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT This proposal describes a five-year mentored physician-scientist training program to define the scope and functional consequences of mitochondrial genome mutations in skin tissue aging. The mitochondrial genome (mtDNA) is an extranuclear source of genetic variability that regulates critical cell functions. MtDNA is highly susceptible to mutagenesis, including the development of large deletion mutations. When a mutation is present, it can affect all copies, or only a fraction of the copies of mtDNA. This portion is referred to as heteroplasmy. Increased levels of mtDNA mutation heteroplasmy have been frequently reported in aging tissue. Despite this potential, the scope and consequences of age-related mtDNA mutations are not well understood due to difficulties in sequencing mtDNA and a paucity of in vitro models, making it challenging to use these mutations as a metric of aging or to develop targeted interventions. To address these limitations, I developed a long-read sequencing method to profile mtDNA and an induced pluripotent cell-based system to study the impact of high mtDNA mutation heteroplasmy in multiple cell types. Aged facial skin contains the highest frequency of mtDNA deletion mutations across tissues profiled and prior work suggests a link between mtDNA and function of skin keratinocyte progenitor cells. Skin thus offers an appealing system to begin understanding the breadth and consequences of age-associated mtDNA mutations. I hypothesize that mtDNA alterations are acquired with aging in a tissue specific pattern and directly contribute to age-associated functional changes. To test this, I will pursue three study aims 1) Map and determine the tissue specificity of mtDNA mutations in aged skin, 2) Quantify mtDNA genetic diversity in age- associated neoplasia, and 3) Determine whether high mtDNA mutation heteroplasmy alters the function of human iPSC-derived keratinocyte progenitor cells. This work will provide foundational knowledge for the study of mitochondrial genome mutations in aging. The K08 will support me to develop advanced skills in the identification acquired genomic variation and the use of pluripotent and progenitor cells in the study of aging in order to become a pioneering physician scientist studying the biology of aging. Dr. Michael Teitell, a world- renowned expert in the study of mitochondrial metabolism and highly experienced and successful mentor, will serve as my primary mentor. Dr. Jonathan Wanagat and Dr. Paul Boutros will offer guidance in the study of mitochondrial genomic variation, Dr. Thomas Rando will offer guidance in the study of progenitor cell aging and Dr. Phillip Scumpia will guide work studying cutaneous tissue, all as members of my advisory committee. A K08 award will enable my carving out a unique, critical niche of multidisciplinary research that blends mitochondrial genomics and regenerative biology at the launch of my career as an independent investigator.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Dental plaque represents one of the most complex microbial communities or biofilms known to affect humans. Oral biofilm-related diseases, e.g. dental carries, gingivitis, and periodontitis, impact a large population of all age groups and continue to impose a huge economic burden due to the lack of effective therapies. The development of dental plaque begins with the attachment of early colonizers to the tooth enamel, generating an adhesive matrix that attracts intermediate and late colonizers. Actinomyces spp. are key early colonizers that play a prominent role in biofilm development by their ability to interact directly with the tooth surface and both early and intermediate colonizers. Therefore, our studies have focused on dissecting the adhesive properties, i.e. pilus and non-pilus proteins, dictating these interactions and their assembly mechanism on the surface of Actinomyces oris, the most abundant Actinomyces species in the human oral cavity. During the past grant period, we identified a transmembrane protein named SafA in A. oris that is genetically linked to the housekeeping sortase SrtA (class E sortase), the conserved transpeptidase enzyme that is central to the morphogenesis and cell wall anchoring of pilus and non-pilus proteins. Specifically, safA deletion results in SrtA cleavage by signal peptidae LepB2 and extracellular excretion of SrtA, concomitant of secretion of non-pilus proteins and most pili, while the remaining cell-wall anchored pili are exceedingly long but unable to mediate bacterial coaggregation. Strikingly, the defects of ∆safA in SrtA membrane localization, cell wall anchoring of pili and surface proteins, and coaggregation can be rescued by ectopic expression of SafA homologs from Corynebacterium matruchotii and Corynebacterium diphtheriae, suggesting the conservation of SafA-associated mechanism in Actinobacteria. Remarkably, the ∆safA mutant is severely defective in coaggregation with Porphyromonas gingivalis (Pg) and in inhibition of Pg growth, in contrast to wild-type cells. Intriguingly, in Bifidobacterium the housekeeping sortase SrtE contains a C-terminal SafA domain, suggesting the co-evolutionary existence of class E sortases and SafA. Therefore, we propose that SafA is an evolutionarily conserved antagonist of signal peptidase that topologically modulates sortase function in polymicrobial interaction and biofilm formation. Using biochemical, genetic, and structural approaches, we aim to test this central hypothesis by elucidating the antagonistic mechanism of SafA-mediated topological modulation of sortase function in polymicrobial interaction and biofilm formation in A. oris, determining the conservation of this SafA-associated mechanism and the co-evolutionary existence of class E sortases and SafA in Actinobacteria, and examining how the SafA-associated mechanism modulates A. oris coaggregation with Pg and Pg virulence. The conservation of sortase-mediated surface assembly and SafA-associated mechanism in Actinobacteria thus magnify the significance of our studies on how SafA topologically modulates sortase function in A. oris that likely affects the virulence potential of other oral pathogens.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Myofascial Pelvic Pain (MPP) is a frequently overlooked musculoskeletal cause of chronic pelvic pain affecting 10-20% of all adult women. Pelvic floor muscle tightening in MPP results in acidosis and accumulation of neuroinflammatory mediators, leading to self-reinforcing pelvic pain and bowel, bladder, and sexual dysfunction. A challenge in the management of MPP has been a lack of quantitative measures of pelvic floor physiology capable of quantifying functional alterations in the pelvic floor musculature in 3- dimensional space. This deficiency has in turn hampered both a greater understanding of the mechanisms of MPP and the development of novel therapeutics. Our research group has developed an innovative approach to identify pelvic floor muscle dysfunction using a near infra-red spectroscopy (NIRS) optical, vaginal interface designed to image and quantify in real-time oxygenation and hemodynamics regionally throughout the pelvic floor in response to contraction and relaxation of the muscle. During the R61 phase, we plan to validate this technology as an effective, non-invasive, and scalable method to assess pelvic floor muscle fitness. We will seek to establish parameters to describe the severity and spatial patterns of myofascial dysfunction and how these parameters correlate with individual muscular assessment, the spectrum of pelvic pain manifestations, and bowel, bladder, and sexual complaints. We will also correlate these findings with current subjective methods of pelvic floor assessment in control subjects and MPP subjects, specifically focusing on the ability of NIRS to provide repeatable measurements independent of user expertise and training. In the R33 phase, we will utilize this technology to examine prospective changes in pelvic floor fitness in MPP patients following a range of myofascially-directed therapies, including pharmacologic muscle relaxation, onabotulinumA toxin pelvic floor injection, and myofascial release physical therapy. We will correlate NIRS outcomes and symptomatic measures with systemic inflammatory markers pre- and post-treatment to understand better the role of neuroinflammation in the clinical syndrome of MPP. We will also employ machine learning models to examine if this technology can be utilized to predict responses to individual treatments. Demonstration that objective measures of muscle fitness recover as symptoms improve following myofascially-directed therapies will provide the first direct evidence of the underlying nature of MPP, evidence needed to evolve current therapeutic approaches. Through interdisciplinary collaboration of specialists in urology, gynecology, integrative medicine, and computational medicine, this novel imaging technique providing both spatial and functional evaluation of the pelvic floor will assist in the diagnosis and management of chronic pelvic pain and promote additional research into novel approaches to address the poorly recognized epidemic of myofascial pelvic pain in women.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT (OVERALL) Autoimmune endocrinopathies include a number of conditions in which the immune system destroys healthy hormone-producing tissues. These conditions affect a large number of Americans, many of them female, but have not yet been the focus of ACE studies in the past. Moreover, unlike most other autoimmune conditions, autoimmune endocrinopathies are not usually treated with immunotherapies, despite clear evidence that their etiology is immune-mediated. Thus, there is an urgent need to better understand autoimmune pathogenesis to identify targets for immunotherapies in these conditions. This new ACE proposal seeks to delineate mechanisms of autoimmune chronicity as critical pathways in the establishment of autoimmune endocrinopathies. The Principal Project delineates the epigenetic and transcriptional mechanisms underlying chronicity in autoimmune endocrinopathies. These studies will determine whether an epigenetic regulator UTX may a role in conversion of stem-like progenitor CD8+ T cells to effectors in autoimmune endocrinopathies. The Collaborative Project addresses spatial and temporal determinants of immune chronicity in cancer immunotherapy-related immune related adverse events (IRAEs). Endocrine IRAEs of great interest given the large proportion of cancer patients who now receive immunotherapies and develop these unwanted side effects. Finally, many autoimmune endocrinopathies are sex-biased, and the Pilot Project addresses sexual dimorphism in immune chronicity pathways. In particular, UTX is an X-linked gene that is differentially expressed in male vs. female CD8+ T cells, and we will explore UTX’s role in mediating sex differences in progenitor to effector conversion. Our ACE includes an Administrative Core, which will ensure efficient day-to-day operational support; an ACE Funds Management Core, which will administer financial and consortium agreements; and an ACE Biorepository Core, which takes advantage of the well-developed biorepository infrastructure system in place at UCLA. Our program will build a cohesive and multi-disciplinary team of immunologists, clinicians, and computational biologists to contribute our deep expertise to the ACE collaborative enterprise.