University Of California Los Angeles

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY/ABSTRACT Immunologically, the enigma of the granuloma is best reflected in that sterile, bacillus-controlling, and progressive granulomas can coexist in the same lung, with the progressive form ultimately killing the host. This observation is consistent with the modern concept of “concomitant immunity: the paradoxical immune status in which resistance to reinfection coincides with the persistence of the original infection”. Here, we will test the hypothesis that the development of concomitant immunity regulates macrophage differentiation, influencing granuloma formation and ultimately the outcome of the battle between the immune response and the pathogen Mycobacterium tuberculosis. To do so, we will use single cell RNA sequencing and spatial sequencing to map the coordinates of cell populations and antimicrobial mediators in human TB granulomas. We propose the following specific aims: 1) elucidate the cellular and molecular architecture of human pulmonary TB granulomas, 2) investigate the role of macrophage subpopulations that contribute to the antimicrobial response vs. pathogenesis of TB granulomas; and 3) investigate the role of T cell subpopulations in contributing to concomitant immunity in TB granulomas. We will identify specific cell subpopulations that contribute to host defense by comparing individual TB granulomas with varying bacterial loads and those with pathogenesis by examining the dynamic change with the progression of primary lesions, to early lesions with bronchial obstruction, to post-primary granulomas. We will determine the role of macrophage subpopulations in host defense and pathogenesis, in particular the foamy macrophages that we discovered express TREM2. We will investigate which T cell populations are predictors of individuals that respond to chemotherapy versus those that are resistant and therefore serve as biomarkers. These studies will bring together collaborators at UCLA, the Ragon Institute and the Institut Pasteur de Tunis with expertise in clinical tuberculosis, immunology and molecular biology to gain new insight into the mechanisms by which concomitant immunity influences granuloma structure to optimize host defense against TB.

NIH Research Projects · FY 2025 · 2022-02

SUMMARY Pulmonary arterial hypertension (PAH) is a chronic lung disease characterized by a progressive increase in pulmonary arterial pressure leading to right ventricular (RV) heart failure and death. Over the last five years, our group and others have demonstrated the critical role of oxidized fatty acids in the pathogenesis of PAH. Levels of oxidized fatty acids of the lipoxygenase (LOX) pathway; hydroxyeicosatetraenoic acids (HETEs) are upregulated in the lungs of patients with PAH as well as in rats with pulmonary hypertension (PH). We were the first to establish that dietary supplementation of a single oxidized fatty acid (15HETE) of the LOX pathway is sufficient to cause PH in wild type mice in the absence of any other PH stimulus. In our very recent publication in Hypertension, we reported T cell-dependent endothelial cell apoptosis as one of the mechanisms underlying 15HETE induced PH. However, the exact molecular mechanisms leading to PAEC apoptosis and onset of PH are not known. To unravel the molecular mechanisms, we performed RNA-Seq on the lungs and intestine of mice on 15HETE diet and integrated our RNA-Seq data with online microarrays of human PAH lungs and identified IFI44 (IFN inducible protein 44) as the only novel common gene that was significantly upregulated. IFI44 is an interferon inducible protein and our preliminary data shows that IFNα4 is specifically increased in the small intestine of mice on 15HETE diet. Our preliminary time course experiments revealed increased expression of IFI44 in the small intestine, which precedes its upregulation in the lung, suggesting that 15HETE may act on the small intestine initially. In addition, our pilot data shows that IFI44 is expressed in the immune cells in the lungs of human PAH patients and in T cells in mouse lungs on 15HETE diet. Our bioinformatic analysis also revealed that expression of IFI44 in immune cells in the lungs correlates with CXCL10 (a proinflammatory cytokine) and TRAIL (Tumor Necrosis Factor Related Apoptosis Inducing Ligand). TRAIL and CXCL10 are both known to induce endothelial cell apoptosis. Our preliminary data also shows i) Knockdown of IFI44 in the lungs of mice on 15HETE diet prevented development of PH, and ii) blocking the pathologic action of Cxcl10 rescues PH development. Taken together, our overall hypotheses are i) dietary 15HETE acts on intestinal epithelial cells to produce IFN4, which induces IFI44 in specific immune cells of the Lamina Propria. IFI44 positive immune cells migrate to the lungs and in coordination with CXCL10/TRAIL trigger PAEC death, causing PH; and ii) IFI44 and CXCL10 can serve as novel therapeutic targets in the lungs to prevent or even rescue development of PH in mice on 15HETE diet. Aim 1 will determine the mechanisms by which 15HETE activates the intestinal epithelium resulting in the development of PH; Aim 2 will examine how activation of IFI44 in immune cells in the small intestine induces endothelial cell apoptosis in the lungs through CXC10 and/or TRAIL dependent mechanisms causing pulmonary hypertension; and Aim 3 will examine whether IFI44 and CXCL10 can serve as novel therapeutic targets in the lungs to prevent or rescue development of PH in mice on 15HETE diet.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY / ABSTRACT Cells of the innate immune system such as monocytes and macrophages can be reprogrammed by their environmental context. Stimuli in the environment, such as cytokines and pathogen-associated molecules, can dramatically alter cellular phenotype. This phenomenon, termed “trained immunity,” is driven by epigenetic changes to the enhancer repertoire. Upon stimulation, transcription factors such as NFκB can bind to closed chromatin regions, open the chromatin, and facilitate histone modification, activating previously silent regulatory DNA elements called latent enhancers. These epigenetically reprogrammed innate immune cells respond differently to subsequent stimulation and alter host responses to infectious diseases. Interestingly, the effects of trained immunity are variable. In some cases, trained immunity primes the host to produce an increased inflammatory response upon secondary stimulation, but in other cases it diminishes the inflammatory response. Whether reprogrammed monocytes produce increased or diminished inflammatory responses depends on what stimulus forms the memory. That is, the effects of trained immunity are stimulus- specific. However, the mechanisms that account for this stimulus-specificity are unclear, and understanding these mechanisms will be critical for harnessing the therapeutic potential of trained immunity. The central hypothesis of this grant is that different dynamic features of NFκB activity contribute to the stimulus- specificity of trained immunity. NFκB is a ubiquitous transcription factor that is considered the master regulator of inflammatory gene expression. Although nearly all pathogens and cytokines activate NFκB, they do so with varying dynamics. In particular, some stimuli induce an oscillatory pattern of activity, while others induce a non- oscillatory pattern of activity. We have shown that non-oscillatory NFκB results in more de novo enhancers than oscillatory NFκB using a mouse model in which NFκB oscillations are perturbed. Aim 1 of this proposal characterizes the stimulus-specificity of training in vivo and tests the hypothesis that trained immunity produced by different NFκB dynamics differentially alters host response to secondary infection with Candida albicans. Aim 2 investigates the mechanisms by which NFκB dynamics-dependent enhancers regulate gene expression. This mentored career development award will support the training of a promising junior faculty member at UCLA. The specific training objectives are to acquire expertise in 1) bioinformatics, 2) mouse models of infection, 3) immune profiling of tissue samples, and 4) the field of innate immune memory. Training will occur through mentorship by senior faculty members, coursework, seminars, and international conferences. The goal of this career development award is to establish the principal investigator as an independent physician-scientist, ultimately producing diagnostic and therapeutic tools for immune-mediated complications of infectious diseases.

NIH Research Projects · FY 2025 · 2022-02

DESCRIPTION (provided by applicant): Most neurological and neurodevelopmental disorders are sex-biased in incidence or severity. To understand why one sex may be vulnerable to a disease, it is important to understand brain development in both sexes. Mood, cognition, and other processes are regulated by estrogen receptor (ER) α neurons. Female have increased number of ERα neurons in the hypothalamus, compared to males. This is an example of a sex difference in neurochemical phenotype, the most common type of sex difference in the brain. Despite this, little is known on the mechanisms controlling their development. Our research suggests that epigenetic mechanisms underlie sex differences in neurochemical phenotype that may contribute to sex biases in disease. Inhibiting DNA methylation in the brains of newborn mice reduces sex differences in ERα in the preoptic area (POA) and the ventrolateral area of the ventromedial hypothalamus (VMHvl) at weaning. We recently reported that DNA methyltransferases (Dnmts; which add methyl marks) and ten eleven translocases (Tets; which remove methyl marks), peak shortly after birth in both sexes in the hypothalamus. Additionally, females have higher expression of Dnmts, while males have higher expression of Tets during this period. This suggests that DNA methylation and hydroxymethylation are dynamic and sex- biased during neonatal brain development. Interestingly, both sexes have an equally high number of ERα cells in the VMHvl at birth, but only in males, it decreases 50% by weaning. This proposal will test the hypothesis that ERα cells in males, but not females, accumulate DNA methylation marks during postnatal development which establishes the sex difference. The F99 phase will test 1) whether subpopulations of ERα cells with functional roles are sensitive to neonatal inhibition of DNA methylation using single-molecule fluorescent in situ hybridization and 2) whether specific sub-populations downregulate ERα expression across development (Aim 2a). Lastly, using methylated and hydroxymethylated DNA immunoprecipitation sequencing, it will test the hypotheses that there are global sex differences and developmental changes in the epigenome, and specifically, that the ERα promoter in males has increased levels of DNA methylation compared to females (Aim 2b). The proposed study will help the candidate, Laura Cortes, achieve her goal of becoming a tenure-track professor at an R1 institution. This proposal will provide training in cutting-edge techniques, such as sm-FISH and epigenomic sequencing, to investigate how DNA methylation regulates neurochemical phenotype in both sexes. The Neuroscience Institute at Georgia State University is an ideal environment given the 1) access to the expertise of reputed neuroendocrinologists and state-of-the-art tools, 2) collaborative intra-departmental and inter-institutional atmosphere, and 3) the plethora of career development opportunities. Completion of the training plan described in this proposal will ensure the development of a well-rounded and successful scientist capable of transitioning into an independent investigator.

NIH Research Projects · FY 2025 · 2022-02

Summary/Abstract Pancreatitis is a potentially fatal disease of exocrine pancreas, with obscure pathogenesis and no specific or efficient treatment available. Excessive/unresolving inflammation is a major determinant of pancreatitis severity but its mechanisms are poorly understood. Recent findings in experimental and genetic models have uncovered key pathogenic roles of impaired autophagy and mitochondrial dysfunction in acinar cell damage driving pancreatitis; disordering of these pathways is also prominent in human disease. Blockade/impairment of pancreatic autophagy initiates inflammation in genetic models, such as pancreas-specific knockout of the key autophagy mediator ATG5. Conversely, enhancing autophagic efficiency (or normalizing mitochondrial function) ameliorates the inflammatory response in experimental pancreatitis. These findings strongly indicate that normal autophagy restricts inflammation in pancreatitis; however, the underlying mechanisms are unknown. We hypothesize that a key mechanism linking impaired autophagy to inflammation in pancreatitis is acinar cell mitochondrial dysfunction; and further, that this mechanism is mediated by the increase in acinar cell mitochondrial (mt)ROS and mtDNA release. We posit that these effectors activate major proinflammatory pathways: NF-κB, triggering the expression of cytokines/chemokines in acinar cells and thus initiating the inflammatory response; the DNA-sensing cGAS-STING-TBK1 pathway in macrophages, resulting in production of type I IFNs; and inflammasome activation in macrophages, resulting in massive production of IL-1β and IL- 18. The proposed studies will use genetic and pharmacologic approaches to elucidate the roles of these pathways. The hypothesis will be tested in 3 Specific Aims: 1). Determine the effects of impaired autophagy on mitochondrial dysfunction and pancreatic inflammation in genetic and experimental models of pancreatitis. 2). Investigate macrophage cGAS-STING-TBK1 pathway activation caused by impaired autophagy in genetic and experimental models of pancreatitis. 3). Investigate the role of (canonical) inflammasome pathway in macrophage activation caused by impaired autophagy and mitochondrial dysfunction. Our proposed studies will determine the mechanisms linking impaired autophagy and mitochondrial dysfunction to macrophage-driven inflammatory response of pancreatitis. The studies will identify molecular targets in these pathways that could be amenable for pharmacologic intervention to alleviate the inflammatory response of pancreatitis.

NIH Research Projects · FY 2026 · 2022-02

Project Summary Aim 1 addresses the dearth of drugs for dementia, by structure-based drug design. This approach, so fruitful for treating cancer and HIV-AIDS, is opening for Alzheimer’s Disease (AD) because of advances in diffraction and cryoEM. Aggregation of protein Tau is strongly correlated with the onset of dementia. From the recent near-atomic structure of Tau fibrils extracted from the autopsied brain of an Alzheimer’s patient, the drug-binding site (or pharmacophore) has been determined for a fibril-disaggregatng compound. By screening compounds that fit the pharmacophore, new Tau disaggregants have been discovered. These disaggregants dissolve AD-brain Tau fibrils, but do not produce toxic products. From further cycles of structure determination of complexes of Tau fibrils with the new disaggregants, followed by compound screening, safe and effective compounds will be sought to reverse the toxic aggregation of Tau in the brain. Synthetic chemist co-Investigator UCLA Prof. Patrick Harran will collaborate to apply a similar approach to discover complexes of disaggregants with brain-penetrant nanoparticles. Aim 2 proposes to fill the vacuum of knowledge of the structures of small aggregates of Tau and beta- amyloid, known as oligomers. Numerous studies of others provide evidence that oligomers are more cytotoxic than fibrils of the same protein. Oligomers of different fibril-forming proteins share structural similarities in that particular antibodies (A11 & M204) recognize them, but not their corresponding fibrils. The transient nature of oligomers has defeated previous attempts to learn their atomic structures, but our lab has recently discovered a monoclonal Fab that extracts fairly homogeneous oligomers of Tau from AD brains and stabilizes them long enough to make grids suitable for cryoEM structure determination. Preliminary micrographs suggest that antibody ligands permit alignment of beta-amyloid oligomers for cryoEM determination of structure, which may serve subsequently for design of inhibitors and disaggregants. Aim 3 proposes tests of AD drugs in “mini-brains” which are grown in the lab of our co-Investigator UCLA Prof. Novitch. These organoids are the size of a BB yet display structure and electrical properties of actual human brains. They are made from human cells and display the cell types and electrical messaging of human brains. Preliminary work shows these mini-brains can be infected with Tau pathology, and now the ability of our various drug candidates to interfere with the spreading and damage of aggregated Tau can be tested in them. If successful, this approach can provide a new avenue for testing Alzheimer’s drugs prior to human trials. For comparison, our inhibitors and disaggregants will also be assessed in a mouse model of tauopathy.

NIH Research Projects · FY 2026 · 2022-01

There is a broad heterogeneity of clinical outcomes after infection with Coccidioides (Cocci) ranging from asymptomatic infection to mild pulmonary disease (“Valley fever”) to a life-threatening, invasive disease called disseminated coccidioidomycosis (DCM). Everyone in the endemic areas is susceptible to this infection, but we have almost no ability to predict who will develop disseminated disease and lack an understanding of why they do so. With nearly 10K reported cases of Valley fever and 200 cases of DCM yearly in California, our state alone spends ~$1B yearly on coccidioidomycosis. Thus, there is an urgent need to better understand DCM to enable better prevention, diagnostics, prognostics, and treatments. Our team's long-term goal is to study the intersection between the virulence programs of Coccidioides spp that maliciously exploit defective immunity, and the dysregulation of genetic and immunological programs of innate and adaptive immunity that allow for severe disease to take hold. Our program will bring together a cohesive and multi-disciplinary team of immunologists, geneticists, computational biologists, fungal microbiologists, and clinicians. Combining deep expertise with synergistic goals will enable breakthroughs. Our consortium includes four Projects and three supporting Cores: Project 1 addresses the innate immune responses to cocci infection that go awry in the first stages of cocci infection; Project 2 addresses the adaptive immune responses to cocci infection that go awry in protecting the host from disseminated disease; Project 3 addresses the genomic basis of cocci disease, from common variants that underlie susceptibility due to ancestry to rare variants that disable host defenses; and Project 4 addresses the contributions of fungal virulence factors in enabling the organisms to evade host immune defenses in some individuals. Our program includes an Administrative Core (Core A) that facilitates communications between investigators, organizes meetings and finances, and runs the Developmental Research Program. We also propose a unified Clinical Samples Core (Core B) comprising two of the largest coccidioidomycosis clinics in California, and a Model Organisms Core (Core C) that will carry out all experiments requiring BSL3 safety measures. These Cores together empower the proposed projects by providing common reagents, human samples, tools, and expertise. Our proposed investigations have the potential to transform our understanding of invasive fungal infections and will restore hope for patients through new approaches to prevent, diagnose, and treat DCM.

NIH Research Projects · FY 2025 · 2022-01

PROJECT SUMMARY Atrial fibrillation (AF) is the most common cardiac arrhythmia occurring in 9% of population older than 65 and is associated with increased morbidity and mortality, notably from embolic stroke, sudden death and heart failure. Although oxidative stress has been implicated in the pathogenesis of AF, detailed mechanistic insights into oxidase activation and its downstream effectors have remained elusive. We have previously identified a correlation between NADPH oxidase isoform 4 (NOX4) and AF in cardiac transplant patients, and a direct causal role of NOX4 in AF development using RNA based acute induction of NOX4 in zebrafish. In preliminary studies, we have shown that AF develops in a novel in-house generated, cardiac-specific NOX4 transgenic zebrafish line, which will be used in Aim 1 to delineate a causal role of cardiac-specific activation of NOX4 in AF pathogenesis together with a novel murine model of AF established in-house (Aim 1). Notably, these mice exhibit spontaneous AF episodes (absent P valves and irregularly irregular RR intervals), as characterized by real time telemetry ECG analyses. Global and cardiac specific knockout mice will be employed to examine a specific role of cardiac NOX4 in AF development (Aim 1). In Aim 2, we will examine whether NOX4-dependent mitochondrial dysfunction and autophagy mediate AF development in both the zebrafish and mouse models, based on preliminary observations of substantial mitochondrial reactive oxygen species (ROS) production in NOX4 overexpressed zebrafish, and significant upregulation of autophagy marker LC3II in the murine model of AF, which was completely abrogated in NOX4 knockout mice. We will employ autophagy inhibitors and mitochondrial ROS scavengers to examine their effects in preventing AF (Aims 2 & 3), via attenuation of mitochondrial dysfunction-autophagy coupling (Aim 2). Changes in autophagy markers of LC3II, Atg7 and Beclin-1 under MitoTempo treatment will be examined (Aim 2). We have innovatively shown that nitric oxide (NO) attenuates NOX4 activation in ischemia/reperfusion. Indeed, in preliminary experiments NO donor treatment was robustly effective in preventing AF in NOX4 overexpressed zebrafish, and the cardiac specific NOX4 transgenic zebrafish. In Aim 2 we will also examine reversal effects of NO donors on AF, and novel molecular mechanisms underlying NO inhibition of NOX4. In Aim 3 we will use patch clamp, live confocal imaging, and dual voltage/calcium optical mapping to examine the electrophysiological and intracellular calcium (Ca) handling targets of NOX4 expression in aged mice, including the intermediate roles of ROS and autophagy. Our preliminary data indicate that these animals exhibit increased phosphorylation of RyR2, which we expect to drive increased sarcoplasmic reticulum (SR) Ca leak, spontaneous SR Ca release and afterdepolarizations. When one considers that these cellular changes occur in the environment of slowed conduction, which we identified using optical mapping, these changes are highly proarrhythmic. Taken together, accomplishments of these studies employing powerful approaches of innovative model organisms and novel genetic strains will no doubt prompt development of innovative therapeutics for AF and postoperative AF.

NIH Research Projects · FY 2026 · 2022-01

SUMMARY/ABSTRACT Despite a tremendous effort in basic science, clinical trials, drug development, and technical advances in surgery and radiation oncology, glioblastoma remains incurable and improvements in overall survival have been marginal. While radiotherapy is still one of the most effective treatment options for glioblastoma, it cannot control the disease over time. This suggests that novel combination therapies are desperately needed to improve radiation treatment outcome for patients suffering from this disease. The studies outlined in this proposal are based on a hypothesis that is backed by our extensive preliminary data and rigorous published data in the literature. The overall hypothesis is that biomarker-based drug selection predicts synergistic lethality of combination therapies in GICs and glioblastoma bulk tumor cell populations, prevents radiation-induced GBM phenotype conversion and allows for individualized optimization of radiotherapy. The three aims of this study will address this aspect of glioma biology using an innovative tool to track GICs and their progeny, while leveraging the unique resources and expertise available at UCLA and the NIH/NCI CTEP portfolio of drugs. Aim 1 will identify compounds in the NCI CTEP portfolio that interfere with radiation-induced phenotype conversion in glioblastoma and develop biomarker profiles predictive of synergistic lethality in combination with radiation. Studies in Aim 2 will optimize combination therapies in vivo. Finally, Aim 3, will use patient avatar studies to validate biomarker-based drug selection in PDX models of glioblastoma.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY Actin-based protrusions endow cells with a vast variety of forms. Cells on the surface of mucosal epithelial tissues, such as the cornea and mouth, often project elongated, wrinkle-like protrusions called microridges, which are arranged on apical surfaces in maze-like patterns. Microridges help these cells retain mucus, thus protecting vulnerable epithelial tissues from infection, abrasion and drying out. Until recently, almost nothing was known about microridge morphogenesis. Since microridge morphologies are distinct from those of better- studied protrusions, studying them could reveal fundamental new principles of protrusion morphogenesis. In recent years, the Sagasti lab established larval zebrafish skin cells as a model for studying microridge morphogenesis. This model enables powerful molecular manipulations and live imaging of morphogenesis. The Sagasti lab’s initial studies revealed several new discoveries, including: 1) The dissection of microridge morphogenesis into four molecularly separable steps, 2) the discovery that myosin contraction in the apical cortex regulates surface tension to permit microridge formation, 3) the identification of Plakin cytolinkers as master regulators of microridge length, 4) the discovery that keratin filaments are integral components of microridges required for their stability and elongation (the first example of intermediate filaments directly contributing to protrusion morphogenesis), and 5) the discovery that cortical myosin minifilaments orchestrate an unusual microridge fission/fusion rearrangement process that facilitates the formation of regular, periodic microridge arrangements on cell surfaces. Collectively, these discoveries have broad implications for how cortical myosin, cytolinkers, and intermediate filaments contribute to protrusion morphogenesis. Building on these achievements and new preliminary data, the Sagasti lab will pursue two future lines of research. First, since cortical contractility creates the biomechanical conditions enabling microridge morphogenesis, they will investigate how cross-linking proteins and cell junctions temporally and spatially tune cortical contractility to promote microridge formation and patterning. Since cortical contractility controls many cellular processes, these experiments will reveal regulatory mechanisms with wide relevance in cell biology. Second, since Plakin cytolinkers are central regulators of microridge initiation and elongation, they will investigate how the structure of Plakins relate to their functional roles in microridge morphogenesis, and use Plakin proteins as handles for identifying new regulatory components of the cytoskeletal scaffold that creates microridge protrusions.

NIH Research Projects · FY 2025 · 2022-01

PROJECT SUMMARY: Vascular calcification (VC) frequently complicates cardiovascular disease. It increases the morbidity and mortality and constitutes a significant obstacle in interventions and surgeries. The vascular endothelium plays an important role in VC. The intimal (luminal) endothelial cells (ECs) contribute to VC by providing osteoprogenitor cells through endothelial-mesenchymal transitions (EndMTs). The adventitial ECs are known to contribute to neo-angiogenesis in diseased vascular wall, but it is unknown whether such ECs support VC and what defines them. A subset of ECs with high expression of the EC marker CD31 and the glycoprotein Endomucin (Emcn) has been found in bone to support bone formation. It is possible that adventitial or other peripheral ECs are recruited to diseased areas to promote calcification. Preliminary experiments, using the Matrix Gla Protein null (Mgp-/-) mouse as a VC model, showed extensive EC involvement in the calcified aorta. We identified two subtypes of ECs in the adventitial vs. the intimal endothelium (referred to as a-ECs and i- ECs). The a-ECs were CD31+Emcm+ and correlated with the severity of the VC, whereas the i-ECs were CD31+Emcn-. The two ECs had distinct transcriptional profiles with stem cell and osteogenic markers in the i- ECs vs. enhanced Notch expression in the a-ECs. Endothelial deletion of Notch1 reduced the a-ECs and limited VC while promoting cartilage formation and survival in the Mgp-/- mice. The bone transcription factor Osterix was expressed in both types of ECs. We hypothesize that a-ECs are distinct from i-ECs, recruited to nascent VC, and susceptible to Notch disruption. We also hypothesize that Osterix is protective of EC lineage. In Aim 1, we will characterize the a-ECs (CD31+Emcn+) and compare to the i-ECs (CD31+Emcn-) in the Mgp-/- model, and correlate with severity of VC and marker expression. We will identify unique markers for the respective ECs, with comparison to bone, using transcriptional profiles from single cell RNA sequencing (scRNAseq). We will test the concept that angiogenesis is required for VC using angiogenic inhibitors. In Aim 2, we will determine the effect of loss or gain of endothelial Notch signaling on the EC subtypes and VC. We will examine the distribution of Notch components in relation to VC, and generate Mgp-/- mice with endothelial- specific loss of Notch1 or the Notch receptor inactivator Fbxw7. We will use the mice to determine the effect on the appearance of the endothelial subtypes, calcification and transcriptional profiles by scRNAseq. We will also apply loss and gain of Notch to ECs in vitro and identify novel Notch targets and networks. In Aim 3, we will determine if Osterix helps maintain EC lineage or promotes calcification in vitro and in vivo using human aortic ECs and inducible endothelial-specific Osterix gene deletion in Mgp-/- mice. We will compare the transcriptional profiles of ECs with and without Osterix by scRNAseq in order to clarify the effect on EC lineage, a-ECs vc. I- ECs, and involved signaling networks. Our results may have a significant impact on the field of VC, in particular on the understanding of the endothelial pathology and involvement in VC.

NIH Research Projects · FY 2025 · 2021-12

PROJECT SUMMARY Trauma affects the vast majority of people (50-89%) during their lifetime, and it can have lasting impacts on not only psychiatric but also cardiovascular health. Both trauma exposure and posttraumatic stress disorder (PTSD)—the quintessential trauma-related psychiatric disorder—have been linked prospectively to increased risk of developing a range of cardiovascular outcomes. However, as described in a recent NHLBI Working Group report, critical knowledge gaps must be addressed before trauma or its psychiatric sequelae might be novel targets for reducing cardiovascular disease (CVD) risk. Indeed, our current understanding is limited by a disproportionate focus on PTSD when examining subsequent CVD risk in trauma-exposed individuals. Posttraumatic psychopathology manifests in heterogeneous ways (e.g., depression, anxiety, substance abuse) that have been linked to elevated CVD risk in non-trauma-exposed samples. Further, despite differences in the prevalence and manifestations of posttraumatic psychopathology and CVD in men and women, few studies have directly examined sex differences in these associations. The field's ability to investigate these questions has been hampered by a lack of rigorous measures of trauma exposure and posttraumatic psychopathology in most existing electronic medical record databases, which have extensive data on CVD outcomes. This study will address these knowledge gaps by harnessing a unique prospective, population-based trauma cohort in order to characterize “cardiotoxic” manifestations of posttraumatic psychopathology in men and women. This trauma cohort was created using Danish electronic health registry (EHR) data as part of R01MH110453 (PI: Gradus), and it identified over 1.4 million individuals exposed to a trauma between 1994 and 2016. This established data source includes rich, highly valid, and complete registry-based data with up to 25 years of follow-up on psychiatric and cardiovascular diagnoses following trauma. We will use this existing resource, restricted to persons age 18 years and older with no prior CVD events (n = 1,068,100), to comprehensively examine posttraumatic psychopathology as a predictor of incident CVD. In Aim 1, we will harness cutting edge, novel data science techniques (machine learning) to identify particularly “cardiotoxic” manifestations of psychopathology after trauma. Given sex differences in psychiatric disorders and CVD, sex-specific psychopathology profiles associated with incident CVD risk will be examined in stratified analyses. In Aim 2, we will use traditional analyses to quantify discovered psychiatric predictors and unanticipated/novel psychiatric comorbidity profiles associated with CVD risk in the machine learning analyses, as well as a priori literature-based combinations of psychiatric disorders that increase CVD risk. This EHR-based trauma cohort provides a unique opportunity to consider comprehensively the constellation of posttraumatic psychopathology that may predict CVD, and the results of this study will be used to ultimately inform the development of targeted CVD prevention efforts in trauma-exposed populations.

NIH Research Projects · FY 2026 · 2021-12

Summary Glioblastoma (GBM), the most common primary brain tumor is virtually always fatal. The primary modes of therapy—surgery, radiation and chemotherapy with temozolomide—have led to only marginal improvements in survival. A hallmark of GBM is their high vascularity. Blood vessels within GBM, consisting of mostly endothelial cells and pericytes, not only play the important role of providing nutrients and oxygen to the tumor, but also provide direct trophic support to the tumor cells and serve as conduits for migration out of the tumor. However, anti-angiogenic therapies directed against tumor vasculature have not been successful. A number of studies have revealed that tumor pericytes and endothelial cells can be derived directly from tumor cells, although tumor-derived endothelial cells are relatively rare occurrences in untreated tumors. The number of tumor-derived endothelial cells is greatly increased in recurrent tumors, suggesting that glioma therapy, such as radiation, could influence this process. Our preliminary studies show that radiation can induce the production of endothelial-like and pericyte-like cells in vitro and in animal models in vivo. These reprogrammed cells are important for the growth of the tumor following radiation in vivo and we have begun to define what factors the reprogrammed vascular cells produce to support the growth of the remaining tumor cells following radiation. Our preliminary data indicate that radiation induces altered chromatin states that allow for reprogramming to occur; a process that is potentially therapeutically targetable through the inhibition of the histone acetyltransferase (HAT), P300. The goals of the current studies are to understand the process of vascular reprogramming (RIR) and to determine how it influences brain tumor biology. Our hypothesis is that vascular reprogrammed cells provide critical trophic support to the remaining tumor cells under the harsh conditions that occur following radiation. First, in Aim 1 we will determine whether therapeutically relevant doses of radiation promote vascular RIR. We will then use cell ablation strategies to validate our preliminary data indicating that radiation-induced reprogramming is important for the subsequent growth of the tumor following radiation treatment using both xenotransplantation and immunocompetent syngeneic mouse models. Next, we will explore the mechanisms by which radiation reprogrammed endothelial-like and pericyte-like cells promote the growth of the remaining tumor, determining what specific factors they elaborate, and whether these factors are responsible for tumor survival and growth following radiation. We will then test the hypothesis that radiation induces the formation of vascular-like cells through modification of chromatin accessibility via augmentation of histone acetylation through the P300 histone acetyltransferase, allowing for access of vascular-specifying transcription factors. Finally, we will use pharmacologic agents to therapeutically target the process of RIR through inhibition of the P300 HAT. These experiments can lead to a new understanding of mechanisms underlying resistance to radiation therapy and open the door to new treatments.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ABSTRACT The University of California, Los Angeles (UCLA) and the Charles R. Drew University of Medicine and Science (CDU) in partnership with the Los Angeles Community College District propose the Cultivating Interest in Research Careers (CIRC) Program. Lack of participation from communities underrepresented in medicine and science (UIMS) in the scientific research workforce is a critical issue affecting the future of research, and ultimately our nation’s health. Access to a high-quality science education required for a research career remains largely determined by socioeconomic class, race/ethnicity, gender and/or national origin. There is a critical need to motivate and equip lower-income UIMS community college students with the academic skill set to critically understand, explore and engage in scientific research, as well as the confidence and communication skill set required for advancement in scientific careers. Early interventions in the undergraduate education timeline are needed to build a diverse and inclusive scientific workforce. The Program’s main components include: 1) A Summer Research Experience, where students attend a one-week boot camp to familiarize themselves with research fundamentals and participate in hands-on research under the supervision of a research mentor with expertise in one of NHLBI’s research focus areas, and 2) A Pathway to Research Careers Continuum, where students attend workshops which will enhance their research and academic skills. Since California Community Colleges are the single largest system of higher education in the country, and with data showing that 80% of students enrolled in Los Angeles Community College district schools are from UIMS communities, the proposed CIRC Program will provide important opportunities for UIMS community college students to receive research training as well as skills necessary to transfer to four-year colleges/universities. The Program’s overarching goal is to increase the number of UIMS students prepared to pursue careers in the biomedical and clinical health sciences. To this end, the Specific Aims are as follows: Specific Aim 1: To engage 14 UIMS community college students each year in a mentored, summer scientific research experience in basic, clinical, or translational research related to CVD within the NHLBI mission. Specific Aim 2: To provide 14 UIMS community college students each year with a year-long faculty and “near- peer” advising program to continue building academic skills as well as provide support in transferring to a four year college/university to study math or science. Specific Aim 3: To evaluate the effectiveness of the CIRC program for UIMS community college students in promoting academic advancement, scientific productivity with presentations and/or manuscripts, completion of an undergraduate degree, and successful application to a graduate program in a science-related field.

NIH Research Projects · FY 2026 · 2021-12

Project Summary Sleep has been identified as a state of heightened neural plasticity, but the rules that govern reorganization during sleep remain unknown. To better identify the conditions under which synapses are pruned during sleep, we have begun to examine responses to neural injury in the fruit fly. After antennal transection, flies acutely increase their sleep for up to one day before returning back to baseline levels. Our preliminary data also show that pre-synaptic active zones are removed more rapidly than plasma membrane after antennal injury, and that sleep deprivation after injury prevents the clearance of pre-synapses from injured olfactory receptor neurons. In this project, we will examine: (1) signals that are generated after injuries to promote sleep; (2) contributions of glial cell types to sleep regulation and synapse removal after injury; and (3) molecular mechanisms that promote synapse removal during sleep. We anticipate that these experiments will provide insight into the role for sleep in response to neural trauma, and begin to investigate the consequences of disrupted sleep on recovery from axotomy.

NIH Research Projects · FY 2025 · 2021-09

Abstract The current opioid epidemic calls for global attention. Opioid use disorder is a chronical condition that requires long-term comprehensive health services, from assessment, treatment, continuous monitoring, to extended care. Medication-assisted therapy, although available as an effective strategy to treat opioid addiction, has been significantly underutilized due to the hard-to-reach nature of people who use opioids (PWUO) and the shortage of addiction specialist. An impetus for expanding and enhancing addiction treatment is to mobilize community-based healthcare agencies and family members of PWUO. The study will take advantage of the existing community health care infrastructure and family support systems in Vietnam to develop and test an intervention to strengthen a continuum of addiction services. The intervention, entitled “Community Care Consortium (CCC),” features community health workers’ joint effort with family members to provide patient-centered, individualized addiction care and support. The intervention will be developed and tested through three phases in three regions of Vietnam (Ninh Binh, Da Nang, and Can Tho). In Phase 1, we will conduct formative studies with community health workers, community representatives, PWUO, and their family members to identify barriers to addiction service utilization and discuss potential strategies to establish a continuum of addiction services. Based on the formative study findings, the CCC intervention and its implementation plans will be developed through workgroup meetings with researchers, community members, and target users. In Phase 2, the CCC Intervention will be piloted in three communes and revised based on acceptability/feasibility data, process evaluation, and feedback from field staff and participants. In Phase 3, a randomized controlled trial of the CCC Intervention will be conducted in 60 communes, which will be randomized to either an intervention condition or a control condition (30 in each condition). A total of 720 PWUO, 720 of their family members, and 180 commune health workers (CHW) will participate in the study. The intervention outcomes on PWUO, CHW, and family members will be assessed with the data collected at baseline, 3-, 6-, 9- and 12-month follow-ups.

NIH Research Projects · FY 2025 · 2021-09

Alzheimer’s disease (AD) is a complex age-dependent disorder. It requires multiple approaches to comprehensively understand at a molecular level in order to develop novel diagnostics and disease modifying treatments. Astrocytes and neurons coexist in the brain and both major cell types are known to contribute to AD. The cellular phase of AD is proposed to comprise feedback and feedforward signaling between diverse brain cells as a link between the initial emergence of molecular pathology (abnormal tau and Aβ) and subsequent disease manifestations. Known glial cell proteins that contribute to this cellular phase are APOE and TREM2, and are associated with significantly increased risk of AD. Moreover, known astrocyte mechanisms include reactivity, which is a complex, non-binary phenomenon with sequelae that depends on context. In the past, most disease related studies have evaluated astrocytes or neurons using assessments of physiology, markers, or with gene expression evaluations. Astrocytes and neurons have not been studied in detail together or with cell-type specific proteomic methods, as proposed here and as requested by the FOA. As a result, despite advances, we have little precise information about the proteomes of astrocytes and neurons during aging in brain areas relevant to AD or in brain regions relevant to specific and defined abnormalities such as seizure activity in AD. Our overarching hypothesis is that astrocytes and neurons display protein dynamics during normal ageing and in mouse models of AD and that these changes reflect signaling between these dominant brain cells during the cellular phase of AD pathogenesis and during aberrant seizure activity and its associated cognitive decline in AD. Aim 1 will characterize cell, brain region, and compartment (plasma membrane versus cytosol) specific proteomic methods for astrocytes and neurons. Aim 2 will determine astrocyte and neuron proteomic dynamics during normal aging in mice. Aim 3 will determine astrocyte and neuron proteomic dynamics during aberrant network activity in AD model mice. Understanding the identities and the extent of cell, brain region, and compartment-specific protein changes for the major brain cell types (astrocytes and neurons) using data-driven unbiased approaches could be foundational and catalytic with regards to new opportunities for translational and mechanistic work.

NIH Research Projects · FY 2024 · 2021-09

Abstract The COVID-19 pandemic has posed an unprecedented threat to the health and well-being of Americans, especially low-income and minority Americans and those in poor health or who lost jobs in the economic downturn. The proposed project will examine the protective effect of access to affordable health insurance for poor and low-income Americans in the setting of major health, economic, and social disruptions to their lives. Access to affordable health insurance differs across states because only 35 states and the District of Columbia have adopted the Medicaid expansion to all working-age adults with incomes below 138% of poverty permitted under the Affordable Care Act. The proposed project has three Specific Aims: Aim 1. To assess the protective effects of the Medicaid expansion on insurance coverage for adults and children following the onset of the COVID-19 pandemic. Aim 2. To assess the protective effects of the Medicaid expansion on access to and use of health care, health care expenditures, and financial stress following the onset of the COVID-19 pandemic. Aim 3. To assess the protective effects of the Medicaid expansion on health outcomes, including physical and mental health, for adults and children following the onset of the COVID-19 pandemic. We will also assess how the protective effects differ for disadvantaged persons including poor and low- income persons, African Americans and Hispanics, persons in poor health, and persons who lose their jobs. We will employ two complementary national surveys—the National Health Interview Survey (NHIS) and the Medical Expenditure Panel Survey (MEPS)—in a phased analysis of data straddling the onset of the pandemic. NHIS covers numerous outcomes, with especially detailed measures of financial stress, children's mental health, and some health behaviors. MEPS has more detailed measures of adults' mental health and health care use and expenditures, and its panel design enables us to use analytic methods that more comprehensively control for subjects' characteristics. However, MEPS data are not available until later. We will use regression analysis to model the study outcomes as functions of individual characteristics; baseline state characteristics; time-varying local area characteristics including pandemic severity, depth of the economic downturn, state policies intended to slow spread of the pandemic, and adherence to social distancing; and interactions between states' Medicaid expansion status and local area characteristics. Our approach will enable us to quantify the protective effects of the Medicaid expansion on the study outcomes and assess the main pathways through which these effects occur. The proposed project will assess the value of access to affordable health insurance for poor and low-income persons during the COVID-19 pandemic, but its lessons will be generalizable to other recessions and public health crises, natural and man-made disasters, and people who experience health or economic setbacks in normal times.

NIH Research Projects · FY 2025 · 2021-09

Project Summary/Abstract P50 UC END-DISPARITIES UC END-DISPARITIES will address the inequitable multilevel factors that promote cardiometabolic disease, including hypertension, diabetes, and complications such as atherosclerotic vascular disease and chronic kidney disease. These conditions are highly influenced by structural disadvantage and disproportionately affect low-income, minoritized, and other marginalized groups in the highly diverse and contiguous region of Los Angeles County (LAC) and Orange County (OC) (combined population over 13M; > 46 states). The Center will fill a gap in the current efforts to advance health equity via a multilevel community-academic partnered approach to improve cardiometabolic related health outcomes in targeted LAC and OC Latino, Black, Asian, Pacific Islander, and American Indian communities, which have some of the highest rates of cardiometabolic risk factors and premature cardiovascular morbidity and mortality in the nation. UC END-DISPARITIES will function through three highly integrated cores and three inaugural center research projects across southern California, all supporting Pilot Awards and Community Catalyst Awards, while leveraging synergies with key community-academic networks, community, public health, and health system stakeholders, regional networks of minority-serving institutions, centers at UCLA and UCI for clinical and translational science, and methodologic leaders with expertise in health equity research, biostatistics, implementation science, and health information technology. The Administrative Core, Investigator Development Core, and Community Engagement Core will implement the following Center objectives: 1) Provide administrative and operational support for all activities and collaborate with the NIMHD Chronic Disease Disparities Coordinating Center; 2) Develop the requisite community-academic research infrastructure to improve the health of the diverse multiethnic communities of LAC and OC through multilevel interventions; 3) Broaden/enhance existing and new partnerships with communities to expand the pool of diverse participants in research and recipients of findings for UC END DISPARITIES and related funded entities; 4) Promote successful training and academic advancement of underrepresented post-doctoral and early career investigators through mentorship, opportunities to participate in community engaged health interventions, and support of pilot projects and subsequent independent cardiometabolic disparities research; 5) Conduct rigorous analyses and partner with community stakeholders to identify community, health system, family, and individual-level correlates of health disparities to inform research methods, health policy, and the design of community and health system partnered interventions to mitigate cardiometabolic disparities; and 6) Contribute to the development, evaluation, and dissemination of valid, reliable, and generalizable tools that can measure and track health outcomes and the social, behavioral, and economic predictors relevant to multiethnic communities at risk for cardiometabolic disease.

NIH Research Projects · FY 2025 · 2021-09

The ability to predict when external events will occur, such as anticipating the actions of a predator or the availability of food, is critical for survival. Converging computational and experimental work suggests that dynamically changing patterns of neural activity, including neural sequences, underlie temporal prediction and temporal processing. It is increasingly clear that timing and temporal prediction are highly distributed computations, however, there has been little effort to systematically contrast and understand the computational tradeoffs between how time is encoded in different brain areas. Furthermore, while converging evidence suggests neural sequences in the striatum play a central role in timing, the mechanisms underlying the generation of neural sequences remains elusive. Critically, it is not known whether neural sequences are actively generated within the striatum or are “driven” by neural sequences present in corticostriatal inputs. We propose to address these major gaps in understanding with a combination of innovative experimental and computational approaches. Our key hypotheses are that: 1) neural sequences in the striatum provide a flexible dynamical regime that allows for temporal scaling, i.e., speeding-up or slowing-down of motor responses, 2) cortical input shapes neural sequence formation in the striatum, 3) local inhibitory circuits serve to refine the quality of these sequences in the striatum, and 4) neural dynamics encoding time are widely distributed throughout the brain but are more accurate in certain areas such as the striatum. Our project is anchored in a two-interval timing task in which mice learn to associate two cues with different reward delays, and has three major aims. Guided by large-scale neural recordings in multiple brain areas we will first develop cortical and striatal recurrent neural network models with the goal of understanding which circuit motifs are best suited to generate neural sequences, and determining which models best capture the experimentally observed activity patterns. Second, we will integrate neural recordings and optogenetic perturbations, together with computational approaches, to determine whether neural sequences in the striatum are driven by cortical input and refined by local inhibition, or in contrast actively generated within the striatum. Third, we will carry out a high-throughput electrophysiological survey of neural activity in multiple brain areas, to identify which areas contain the most accurate temporal codes as well as the potential computational tradeoffs between different codes. RELEVANCE (See instructions): By integrating advanced computational and experimental approaches, this collaborative project will provide fundamentally new insights about how the mammalian brain is able to predict when external events will occur, enabling animals to produce appropriately timed movements that are critical in daily life. This work will reveal which brain circuits are most strongly implicated in timing, which is often impaired in neurological disorders such as Parkinson’s and Huntington’s disease.

NIH Research Projects · FY 2024 · 2021-09

Research Summary/Abstract The sheer genetic heterogeneity and complexity of cancer has been laid bare by next-generation sequencing studies of cancer genomes. A major challenge in the post-genomic era of cancer biology is to move beyond descriptive analyses of genomic data and derive functional insights into the complex multigenic interactions that produce complicated phenotypes associated with cancer evolution. Available experimental systems are poorly-suited to address this problem in a systematic and expeditious manner as they generally lack scale, throughput, economy, or biological relevance. We therefore propose the development of a next-generation functional cancer genomics assay that overcomes these limitations and will be employed here to define complicated genotype-to-phenotype relationships in contexts related to prostate cancer initiation and progression. The assay combines the efficient delivery of random, compound genetic perturbations from barcoded lentiviral libraries encoding gain-of-function and/or loss-of-function events; biological selection for a cancer phenotype; and single-cell sequencing analysis to enumerate lentiviral barcodes for the massively parallel association of genotype with phenotype. The major goal of this project is to apply and advance this pioneering strategy to broadly advance our understanding of prostate cancer biology by characterizing compound genetic interactions that (1) transform benign prostate epithelial cells to cancer and establish distinct cancer subtypes, (2) induce neuroendocrine transdifferentiation of prostate cancer, and (3) promote metastatic dissemination. If successful, these studies will characterize and prioritize the functional contributions of multigenic networks to clinically relevant prostate cancer phenotypes. We will also establish multiple new, genetically-defined mouse models of prostate cancer that will better recapitulate the genetic complexity of the human disease. Finally, we anticipate that the next-generation functional cancer genomics strategy will be a flexible and widely applicable experimental tool to rapidly interrogate complex genetic interactions and their impacts on phenotype in many other cancers.

NIH Research Projects · FY 2024 · 2021-09

Abstract Protein interactions, such as protein-protein, protein-nucleic acid, protein-metabolite, and protein-xenobiotic, play a critical role in defining the uniqueness and complexity of biological organisms. Understanding where and when interactions occur is an essential step to functionally characterize the interactome. However, and despite remarkable advances in computational and proteomic technologies, it remains surprisingly difficult to precisely pinpoint contact sites cell-wide in a high-throughput manner. Here we present a new approach, termed Fast Photochemical Oxidation and Capture by Suzuki (FPICS), to map protein interaction sites at high resolution. The key innovation of our method, which represents an unprecedented technical advance, is the use of a single halogen atom as both a photoactivatable molecular 'calling card,' to indicate where interactions occur, and a capture handle, for mass spectrometry-based proteomic detection of each interaction site. With FPICS, halogen substituents are first transferred, using excimer laser irradiation, from halogenated small molecules (e.g. drugs or natural products) or halogenated biomolecules (e.g. proteins, lipids, glycans, oligonucleotides, or metabolites) to interacting proteins. Labeled sites are then captured and identified using bioorthogonal Suzuki–Miyaura cross- coupling chemoproteomic methodology pioneered by our group. FPICS is groundbreaking because it eliminates challenges associated with deconvolving the spectra of crosslinked peptides and the frequent and unwanted fragmentation of large biomolecules. Showcasing the method's wide-ranging applications, here we will apply FPICS map the protein interaction sites for small molecules, lipids, and nucleic acids, aiming to identify new functional and therapeutically relevant binding sites proteome-wide. Taken together, this study will yield a systems-level portrait of the protein interactome, which will lay the foundation for an improved global understanding of the functional significance of the millions of interactions occurring within every cell. The impact of our methods will be wide ranging, spanning the fields of chemical biology, analytical chemistry, and systems biology.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Antimicrobial resistance (AMR) is a global health emergency. Neisseria gonorrhoeae is a high-priority pathogen for AMR, as there are 87 million infections per year worldwide and the bacteria has demonstrated resistance to every class of antibiotics recommended for its treatment. Treatment failures have been reported, typically involve oropharyngeal infections, and are dire warnings that the era of untreatable gonorrhea is near. In the Western Pacific Region, which includes Vietnam, resistance to ceftriaxone and azithromycin, the most commonly recommended antibiotic therapy for gonorrhea, are increasing. Men who have sex with men (MSM) are disproportionately affected by AMR in N. gonorrhoeae infections, but remain under-represented in data from low- and middle-income countries (LMICs). The proposed study seeks to better understand AMR in N. gonorrhoeae by embedding new research into an existing HIV pre-exposure prophylaxis (PrEP) program for MSM in Vietnam. The study aims are 1) to determine the prevalence of and risk factors for AMR in N. gonorrhoeae among MSM in a PrEP program in Vietnam; 2) to measure the frequency of pharyngeal N. gonorrhoeae treatment failures and the association between pharyngeal infections and AMR; and 3) to use whole-genome sequencing (WGS) to conduct a genomic epidemiology study of N. gonorrhoeae nested within our study population of MSM in a PrEP program in Vietnam. The study will leverage regular testing for N. gonorrhoeae and follow-up that occur through the PrEP program to shed new light on the issue of AMR in N. gonorrhoeae within this key population. This Fogarty International Research Scientist Development Award (K01) is to support the career development of Dr. Paul Adamson, an infectious diseases physician whose goal is to become an independent global health investigator at the intersection of AMR and sexually transmitted infections (STIs). The K01 will support Dr. Adamson to develop expertise in 1) clinical microbiology and AMR testing, 2) WGS, bioinformatics, and genomic epidemiology, and 3) advanced training in clinical trials research with a focus in LMICs. To achieve the proposed research and training aims, Dr. Adamson has assembled a mentorship team with proven experience mentoring early-career investigators and with expertise in STIs, epidemiology, AMR, biostatistics, and genomics. Drs. Pamina Gorbach (UCLA) and Le Minh Giang (Hanoi Medical University, HMU) will serve as Primary Mentors and have extensive experience in conducting large-scale prevention research studies and clinical trials on STIs and HIV among MSM. In addition, his Co-Mentorship team includes Dr. Jeffrey Klausner, a US-based global health researcher and an international expert on STIs and AMR in N. gonorrhoeae, and Dr. Nguyen Vu Trung, a Vietnam-based researcher with expertise in clinical microbiology and antibiotic susceptibility testing in N. gonorrhoeae. The proposed research and training aims leverage the strong research collaborations and the robust research infrastructure that exist between UCLA and HMU. The IRSDA will advance Dr. Adamson's career as an expert in STIs and AMR and establish him as an independent global health investigator.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Cell reprogramming represents a major advancement in biology, and has wide applications in regenerative medicine, disease modeling and drug screening. During cell reprogramming, cells experience epigenetic changes that result in a cell phenotype switch. However, whether and how biophysical factors regulate cell reprogramming through epigenetic modifications are not well understood. We have found that biophysical factors, specifically extracellular matrix (ECM) stiffness, has profound effects on epigenetic state and the conversion of fibroblasts into induced neuronal (iN) cells, with the highest efficiency at an intermediate ECM stiffness, which is regulated by focal adhesions and the cytoskeleton. In addition, we have discovered that actin assembly and transport into nucleus plays an important role in epigenetic modulation. Based on our preliminary data, we hypothesize that (1) biophysical cues such as ECM stiffness regulates FAs, actin assembly/disassembly, nuclear transport of actin, and thus, HAT activity to modulate the epigenetic state and cell reprogramming process and (2) an intermediate level of stiffness is optimal for epigenetic remodeling and cell reprogramming. To test our hypothesis, we propose three Specific Aims: (1) Investigate how matrix stiffness regulates iN reprogramming through FAs and actin cytoskeleton, (2) Elucidate how matrix stiffness modulates HAT and the epigenetic state to turn on neuronal genes during iN reprogramming, and (3) Determine the role of actin nuclear transport in matrix stiffness- modulation of HAT and epigenetic state during iN reprogramming. We have assembled a multidisciplinary team with expertise on mechanobiology, cell engineering, high throughput genomic and epigenomic analysis, and live cell imaging to work together and investigate the underlying biophysical and biological mechanisms. Our proposed studies will be one of the first to elucidate how ECM stiffness regulates transcriptomic and epigenetic changes for cell reprogramming, and how ECM stiffness modulates focal adhesions and the cytoskeleton for cell reprogramming. Findings from this project will unravel new mechanisms of cell fate determination, which will have wide applications in cell and tissue engineering, disease modeling and drug discovery, and provide a rational basis for the optimization and development of novel biomaterials for somatic cell reprogramming.

NIH Research Projects · FY 2026 · 2021-09

PROJECT SUMMARY: HIV incidence doubles during pregnancy and postpartum period compared with non- pregnant women, underscoring the urgent need for prevention interventions tailored to high-risk pregnant and breastfeeding women. Incident maternal HIV infections lead to an estimated one-third of all infant HIV infections. South Africa expects over 76,000 infant HIV cases in the next decade; one-third of those can be prevented by eliminating maternal HIV acquisition. Pre-exposure prophylaxis (PrEP) in pregnancy and breastfeeding is safe and effective at preventing HIV. However, PrEP use remains low in pregnancy, and drops precipitously in the postpartum period. Our team is made up of experts in epidemiology, behavioral science, health economics and HIV prevention in pregnant women from University of California Los Angeles and the University of Cape Town. Together we have implemented one of the first studies to integrate PrEP into antenatal care in South Africa (“PrEP-PP”; R01MH116771). We propose to test a novel strategy to optimize PrEP in pregnant and postpartum women in South Africa. Our randomized control trial (RCT) is designed to address key barriers to maternal PrEP use and evaluate cost-effectiveness to inform national policy. This trial builds on our earlier work demonstrating the acceptability, feasibility, safety and potential efficacy of a package of interventions including PrEP, HIV self-testing (HIVST) for the participant and her partner(s), and enhanced adherence counseling (with bio-feedback on adherence) combined with home PrEP delivery for women who want to take PrEP but have difficulties adhering. We developed and piloted a novel intervention entitled, Stepped Care to Optimize PrEP Effectiveness in Pregnant and Postpartum women (SCOPE-PP) that addresses barriers to taking daily PrEP by reducing clinic visit frequency and empowering women to adhere to PrEP. We will evaluate SCOPE-PP in a pragmatic RCT of pregnant and postpartum women at risk of HIV acquisition. We will enroll 500 pregnant women in antenatal care and follow them through 12m postpartum. Women in the intervention will be offered: HIVST for participants & partners, rapid PrEP collection with enhanced adherence counseling. Women with poor PrEP adherence will be offered a differentiated care model of home PrEP delivery to de-link PrEP from clinical visits. The primary outcome is PrEP continuation and adherence in postpartum women, measured through drug levels of tenofovir diphosphate. We aim to: 1) Evaluate the efficacy of the SCOPE-PP intervention on PrEP adherence in pregnant and postpartum women in a RCT; 2) Assess the acceptability and feasibility of integrating SCOPE-PP into ante- and postnatal care using a consolidated framework for implementation research; and 3) Estimate the incremental cost effectiveness of SCOPE-PP vs. standard of care per HIV infection and disability-adjusted life-year averted. This research is critical to inform maternal PrEP interventions to eliminate HIV acquisition and transmission in South Africa and beyond.