University Of California Los Angeles

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY (See instructions): Autism comprises a class of developmental disorders characterized by significant social, communication, and behavioral challenges. Autism is thought to result from neural cell type imbalance during early development, partly from the discovery of chromatin regulators as genes linked with autism. Yet, the cellular, molecular, behavioral, and developmental mechanisms of these autism-linked genes are not well known. This is underscored by the wide variation in type and severity of symptoms. Systematic dissection of the roles of candidate histone modifier autism-linked genes is therefore fundamental to understanding how mutations in these genes leads to cell type imbalances and altered behaviors, affecting physiological well-being. We have taken advantage of the fast-developing vertebrate system zebrafish, in which histone modifier genes are highly conserved, to identify behavioral and developmental phenotypes in mutants of candidate autism-linked genes. By screening for behavioral phenotypes in zebrafish morphants, we have prioritized 7 zebrafish lysine methyltransferase genes (corresponding to 5 human genes) for further study: kmt2a, kmt2ca/b, kmt2e, setd1 a, and setd1 b. Despite their overlapping functions as H3K4 lysine methyltransferases, mutations in these genes led to different behavioral phenotypes in morphants. The full developmental and behavioral phenotypes, the cell types and circuits/pathways, and gene regulatory functions that are affected in these mutants, remain unknown. The proposed study will combine behavioral and developmental assays, pharmacological profiling, brain activity assays, and single-cell transcriptomic and chromatin accessibility profiling to directly test the hypothesis that these genes function to specify cell types required for cell type and circuit development and, ultimately, behavioral responses during development. Altogether, findings from this study will uncover unique functional roles and mechanisms for conserved candidate histone modifier autism-linked genes during early development.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Glioblastoma (GBM) is the most common primary malignant brain tumor with a dismal prognosis that lacks effective therapeutic strategies. While the advent of immunotherapy has significantly improved patient outcomes in a variety of extracranial cancers, these novel approaches have achieved limited success in GBM. The goal of this study is to investigate mechanisms underlying brain microenvironment-specific suppression of effective anti- tumor immunity mediated by aberrant epidermal growth factor receptor (EGFR) signaling, and furthermore whether targeting this oncogene creates novel vulnerabilities to enhance immune response in this deadly disease. Extensive preliminary data, which leverages a highly innovative EGFR activated immunocompetent murine glioma model (MADR-mEGFRvIII), indicate tumor-intrinsic EGFR signaling impairs intratumoral T cell infiltration and promotes the development of a tumor microenvironment (TME) enriched in microglia, a myeloid cell type unique to the brain. Moreover, preliminary data demonstrate an association between EGFR activation and an immunosuppressive wound healing signature linked with poor prognosis in the myeloid compartment of immune cells isolated from newly diagnosed GBM patient tumors. Specific Aim 1 utilizes a transgenic T cell system engineered to be antigen-specific in conjunction with multiple modalities of in vivo EGFR ablation to determine the precise impact of aberrant EGFR signaling and tumor-programmed myeloid cells on T cell cytotoxic function, clonal proliferation, and exhaustion. Through the incorporation of functional and phenotypic assays designed to isolate the contributions of each factor in the complex brain immune TME, this approach is expected to provide novel insight into the role of EGFR activation in dysregulating effector T cell responses. Specific Aim 2 endeavors to elucidate the axis by which aberrant EGFR signaling promotes a distinct immunosuppressive microglia phenotype characterized by VEGF and IL-1β in the brain TME with an emphasis on EGFR-mediated tumor-microglia crosstalk. Using in vitro co-culture approaches and in vivo knockdown strategies, the precise impact of EGFR activation in coordination with microglia IL-1β signaling on tumor- associated microglia phenotype and overall anti-tumor immune response will be assessed. Specific Aim 3 focuses on assessing rational combinatorial strategies targeting EGFR using ERAS-801, a novel clinical stage small molecule inhibitor, to drive enhanced T cell infiltration followed by immune checkpoint blockade to support prolonged T cell function within the TME. Overall, this project will provide the first comprehensive assessment of the mechanistic underpinnings and functional consequences of aberrant EGFR activation on the myeloid- dominant tumor immune microenvironment and the resultant impact on anti-tumor immune response in GBM. By identifying the dynamics of interplay between oncogenic EGFR signaling and immune evasion, this work will not only expand fundamental knowledge on the intersection of these two hallmarks of cancer, but also enable development of near-term therapeutic strategies for patients with this grave malignancy.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Systemic sclerosis (SSc) is a rare autoimmune fibrosing disease that affects 24.4/100,000 people in the United States. While rare, SSc has the highest mortality of all rheumatologic diseases. The complication most associated with this high mortality is interstitial lung disease (ILD). Despite this, there remain limited treatments for systemic sclerosis associated interstitial lung disease (SSc-ILD), and the treatments that do exist only slow but do not reverse ILD. To create new and better treatments for SSc-ILD, we need to better understand the mechanisms that drive fibrosis. A previous study showed that myofibroblasts (the cells that drive fibrosis through the production of extracellular matrix) are derived from lipofibroblasts in a mouse model of interstitial lung disease. Thus, the lipofibroblast to myofibroblast transition in systemic sclerosis may represent a novel target for treatment. To date, however, the mechanisms involved in this transition are not well understood. One known player in the lipofibroblast to myofibroblast transition is PPARg (peroxisome proliferator activated receptor gamma), a master regulator of lipid metabolism and adipogenesis. PPARg has been shown to be downregulated during the lipofibroblast to myofibroblast transition and PPARg agonists reduce fibrosis in mouse models of ILD. In this proposal, we aim to define the mechanisms through which PPARg is itself regulated allowing for the lipofibroblast to myofibroblast transition to occur. We will do this by using a multiomics approach to examine the expression and chromatin accessibility of two genes, PER3 (period circadian protein homolog 3) and CD36 (cluster of differentiation 36), that we believe function upstream and downstream of PPARg respectively. In Aim 1 of this proposal, we will identify the transitional cell stage during which lipofibroblasts are transitioning to myofibroblasts and in Aim 2, we will characterize the role of PER3 and CD36 in the lipofibroblast to myofibroblast transition. Based on previous studies that have defined their roles in adipogenesis (PER3) and fibrosis (CD36), we hypothesize that PER3 inhibits PPARg transcriptional function during the lipofibroblast to myofibroblast transition and that downregulation of PPARg during this transition leads to decreased CD36 expression. Importantly, the use of multiomics allows for an unbiased approach so that we are not limited to exploring changes in gene expression and chromatin accessibility during this transition in one candidate at a time. The proposed research builds upon my background in lipid metabolism research during my time on the R38. The experiments outlined in this proposal will take place at UCLA where I will have access to necessary equipment and core facilities such as the UCLA Tissue Pathology Core Laboratory and the UCLA Technology Center for Genomics and Bioinformatics to support the successful completion of my proposed research. In addition, through my career development and training plan, I will develop new skills in analyzing single-cell RNA-sequencing and ATAC-sequencing data. I will also be able to use the results of this study as a basis for my future K08 application, which I plan to submit in the fall of 2025.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY Better understanding is needed regarding the mechanism of HIV pathogenesis to the developing brain in individuals receiving longstanding antiretroviral therapy (ART). Despite the use of ART, brain abnormalities occur in people living with HIV, especially among individuals with perinatally-acquired HIV (pHIV) where abnormalities are frequently prevalent. Myelin water imaging (MWI) is generally regarded as the most rigorous approach for noninvasive, in-vivo measurement of myelin content. Noninvasive neuroimaging tools can assess the effectiveness of ART in preserving brain health and possibly lead to improvements in therapy. During the past three decades, it has been demonstrated that one-dimensional (1D) MR Spectroscopy (MRS) enables the study of only selected cerebral metabolites due to limited spectral dispersion even with 3 Tesla MRI scanners. Recently, our group, using the home-developed two-dimensional (2D) localized correlated spectroscopy (L- COSY) sequence combined with the prior-knowledge fitting (ProFit) algorithm showed that several cerebral metabolites can be quantified non-invasively in the prefrontal dorsolateral white matter region of perinatally HIV- infected youth and healthy children including novel metabolites such as glutathione (GSH), aspartate (Asp) and scyllo-inositol (sI). However, there were limitations due to the requirement of a 27 ml voxel and longer acquisition times. Hence, a novel five-dimensional (5D) echo-planar J-resolved spectroscopic imaging (EP-JRESI) sequence was recently implemented by our group where two spectroscopic dimensions were combined with three spatial dimensions to study the cerebral metabolite changes in pediatric HIV, with findings reported recently (5R21NS086449-02). However, a total duration of approximately 25 minutes of scan time was necessary. Using a modified rosette trajectory that densely samples both central and peripheral k-space, further acceleration and robustness to motion can be accomplished in MRSI and MRI as compared to other non- cartesian trajectories like radial, which will minimize patient discomfort by significantly reducing the scan time. Hence, the proposed study will test the following hypotheses: 1) The accelerated 5D rosette trajectories-based J-resolved Spectroscopic Imaging (ROSE-JRESI) and MWI with compressed sensing (CS)- based reconstruction will shorten the total acquisition duration, improving its clinical applicability while the 5D ROSE-JRESI data will also offer improved spatial resolution. 2) In the brains of pHIV young adults, decreased axonal myelination is present even in virally suppressed individuals correlating with worse neurocognitive performance (gross motor impairment) and altered metabolites. We propose two specific aims: 1) To optimize 5D ROSE-JRESI and 3D MWI on a 3T MRI scanner, and further optimize the CS-based reconstruction of the 5D MRSI and 3D MWI data using home developed MATLAB codes; 2a) To acquire multi-voxel 2D J-resolved spectra and 2D spectroscopic images of different cerebral metabolites in perinatally-HIV-infected and healthy adults (ages 20- 40 years); 2b) To record myelin water fractions of the brain and to correlate MWI measures with metabolite ratios, neuropsychological test scores and other HIV disease variables.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY There is a need for the development of objective measures of intraocular inflammation to better detect and monitor uveitis. This proposal seeks funds to support early career researchers and trainees to attend the UCLA / American Uveitis Society (AUS) Second International Workshop on Objective Measures of Intraocular Inflammation for Use in Clinical Trials. The goals of the 2024 Workshop will be to review the proposed outcome measures of intraocular inflammation, how these measures relate to clinical trial design; and the potential techniques for objective quantification of study variables. We will also review the progress toward standardization and validation of those techniques over the five years since the 2019 UCLA/AUS workshop and to discuss plans for future studies that will advance this field further. These goals will be addressed through our Aims by providing a dynamic scientific venue for the discussion of evaluation of intraocular inflammation, supporting new collaborations and discussions across a multi-disciplinary attendee population, and by fostering the career development of early career researchers and trainees. This Workshop will feature a diverse roster of attendees of ophthalmologists who participate in clinical research dealing with inflammatory eye diseases (uveitis specialists, retina specialists, pediatric ophthalmologists); clinical trialists; and pharmaceutical and device industry representatives. With this Workshop, we will also be addressing one of NEI’s Bold Predictions, “Infrastructure for large-scale sharing and analysis of vision-related data, including definitions and standardization of data elements and biomarkers across multiple data types, will enable knowledge discovery and predictive disease modeling”. We will also address the NEI’s Area of Emphasis of the Immune System and Eye Health by evaluating imaging biomarkers for disease surveillance and detection. By promoting the multi- disciplinary exchange of ideas and by supporting early career researchers and trainees, we anticipate this Workshop will generate new collaborations and research opportunities for the next generation of investigators. In turn this will lead to further development of objective measures of intraocular inflammation, resulting in earlier detection and improved monitoring of uveitis, which may prevent permanent vision loss and disability.

NSF Awards · FY 2024 · 2024-08

With the support of the Macromolecular, Supramolecular and Nanochemistry Program in the Division of Chemistry, Prof. Yu Huang of the University of California, Los Angeles, will conduct systematic studies to understand and develop high-performance electrocatalysts for hydrogen chemistry. The hydrogen evolution reaction (HER) and hydrogen oxidation reaction (HOR) are central to the hydrogen cycle and the hydrogen economy. This project aims to investigate how the local chemical environment modifies the local reactant supply and fundamentally alters the reaction pathway and kinetics. The knowledge gained from the project will aid the development of more efficient and longer-lasting catalysts for green hydrogen production or more economical hydrogen fuel cells. It will broadly benefit clean energy technologies, including renewable fuels, batteries, and fuel cells. The project will also offer valuable educational and training opportunities for next generation of workforce in renewable energy, with efforts to recruit female students and those from underrepresented groups. This project will systematically tailor the local chemical environment on the Pt surface via transition metal decorations and explore their role in modifying hydrogen chemistry kinetics. The specific tasks include: developing general strategies to modify Pt catalysts with a series of transition metal (Pt-TM) or transition metal oxide species (Pt-TMOx); conducting detailed compositional and structural analysis to identify local chemical and bonding structures; performing systematic electrochemical studies to quantitatively compare HER/HOR kinetics of Pt catalysts with different TM/TMOx decorations; conducting in-situ characterizations to probe local chemical structures, surface adsorbates, and the near-surface environment for Pt catalysts with different TM/TMOx decorations, and establishing their correlation with HER/HOR activity; through existing collaborations, conducting theory calculations to understand the interplay among surface decorations, local chemical environment, and HER/HOR kinetics, and developing the underlying framework for designing future catalysts with vastly improved activity and durability. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

This Research Advanced by Interdisciplinary Science and Engineering (RAISE) award is made in response to Dear Colleague Letter 23-109, as part of the NSF-wide Clean Energy Technology initiative. Heat losses and gains through windows account for 25-30% of overall building energy heating/cooling needs, in turn corresponding to 5-6% of the ~100 Quad of energy consumed in the U.S. every year. Optimizing heat flows through windows, as well as opaque walls, could thus have a profound impact on energy efficiency and decarbonization in the face of climate change. This project enables the synthesis of new materials that will tackle a significant fraction of this energy usage by more effectively managing the flow of radiant heat, particularly in urban heat island scenarios, thereby improving energy efficiency as well as heat resilience. Existing technologies focus primarily on controlling solar heat gain through different components of the building envelope (e.g., roofs, walls, windows, and skylights). However, the buildings absorb heat from its immediate environment as well as emit it to the cold overhead sky via radiation at long-wave infrared (LWIR) wavelengths. This research program at the University of California – Los Angeles leverages this ubiquitous heat exchange to enhance building efficiency by tailoring the spectral and directional characteristics of infrared emissivity. The researchers investigate new fundamental mechanisms based on designed porous materials that can control thermal emission over long-wave infrared wavelengths. At a community level, the team engages particularly vulnerable communities on the impacts of extreme heat to better understand potential energy impacts of the developed classes of materials as well as the materials’ design. Additionally, through this project, the researchers conduct STEM outreach to school students in the area, develop a new high school-level experiment on structural color, and create a short video on the impact of extreme heat and the role of materials to engage a global audience in understanding the potential of new advancements to make our built environment more efficient and resilient. The project enables the synthesis, characterization, and demonstration of new classes of mesoporous photonic metamaterials that are capable of achieving highly tailored control of their spectral emissivity over infrared wavelengths. The researchers study their directional characteristics with a view to their utilization for building-scale control of radiative heat transfer and to more effectively exploit radiative cooling. They investigate the design and synthesis of spectrally selective aerogel-based metamaterials, leveraging the deep-subwavelength nature of introduced porosity to enable previously inaccessible emissivity profiles over infrared wavelengths. To broaden the space of spectral and directional selectivity, they investigate three dimensional Parity-Time (PT) symmetric metamaterials, built using nanoporous colloidal crystals that exhibit highly anomalous directional response in their infrared emissivity. Finally, the team demonstrates that the developed mesoporous metamaterials can deliver the radiative heat transfer benefits needed for building-scale applications. To that end, they develop computational tools to generate digital twins of the ordered or fractal meso¬porous structures to efficiently solve electromagnetic wave transport therein, accounting for dependent scattering, coherent back-scattering, and near field effects, as well as absorption. The project also includes experimentally demonstrating potential impacts on energy efficiency of the developed metamaterials through outdoor testing in simulated urban heat island scenarios. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Pancreatic cancer is an aggressive, painful malignancy with a 5 year survival rate of less than 5% and median survival of about 4 to 24 months. The overall lack of success of chemotherapy treatments underscores, at least in part, the incomplete understanding of how pancreatic adenocarcinoma (PDAC) grows and invades local and distant tissues. There continues to be novel discoveries with respect to genetic mutations that predispose individuals to developing PDAC, histopathological features with prognostic implications, and clinical imaging (functional metabolic and anatomic, e.g. PET/CT) that define clinical stage and guide treatment options. However, overall, each diagnostic modality in large part, remain independent of one another. Consequently there is a need to assess and integrate measurements from the cell, tissue, and organ levels with multiple imaging modalities and to evaluate the concordance across different types of measurements and spatial scales in order to characterize the “molecular omic-type to clinical phenotype” relationship. Linking measurements from these different disciplines, radiology, pathology, and high-throughput molecular measurements, will help to establish more definitive phenotypes of PDAC that are consistent across multiple spatial scales. Recently many of the changes in PDAC have evolved around changes in the metabolic aspects of the tumor microenvironment and the interactions between tumor cells and the surrounding stromal cells, thus we focus on identifying metabolically driven/related phenotypes. We hypothesize that characterizing the PDAC microenvironment tumor-stroma interface at the cell and tissue levels will enable assessment of glycolytic and inflammatory characteristics of tumors in relation to specific histological tumor characteristics (e.g. perineural invasion and lymphovascular invasion) and this will lead to 1) a non-invasive imaging test to identify PDAC phenotypes and 2) new potential cellular targets for treatment. A data-driven systems biology approach using transcription-signaling-metabolic network reconstructions and constraint-based modeling will be used as the tie that binds a wide array of disparate data, including cellular microscopy, 3D preserved architecture tissue imaging, and PET/CT imaging in order to define a non-invasive functional, metabolic phenotyping measurement of PDAC that corresponds to cellular phenotypes. The constraint-based genome scale integrative modeling framework allows the simultaneous analysis of multiple types of data in interacting populations of cells. Additionally through the acquisition of multi-parametric cross- sectional imaging, it will be possible to evaluate whether or not metabolic changes on the cellular level have any manifestations on the tissue level. The environment, team, and resources assembled at UCLA are poised in a unique position to carry out these studies and achieve success in developing a non-invasive imaging approach for phenotyping PDAC.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT The oral mucosa is constantly exposed to a bacteria-rich environment coupled with continual physical trauma. For patients, chronic oral wounds aren't merely an inconvenience. They can impair nutrition intake and pose a substantial risk of debilitating infections, marking a significant oral health challenge. Defining factors involved in oral wound healing would allow the identification of novel therapeutic targets to improve tissue repair. Oral wound healing has long been considered a model of optimal wound resolution characterized by rapid and scarless wound healing. However, the intrinsic genetic and epigenetic controlling mechanisms that make oral mucosa highly adaptable to accelerated healing upon injury remain unclear. An important aspect of healing is re- epithelialization, which entails both proliferation and migration of keratinocytes at the periphery of the wound. Understanding the role and function of keratinocytes in re-epithelialization will benefit patients with chronic wounds. We recently identified a previously unnoticed landscape of non-coding RNA in saliva and found piwi-interacting RNA (piRNA) is surprisingly abundant in saliva. Of particular interest are three distinctive oral-piRNAs that are uniquely present in saliva and oral keratinocytes and appear to contribute to the oral wound re-epithelialization phenotype. The preliminary data involving inhibition of oral piRNAs concomitant with a failure of wounds to heal in a timely fashion suggests that these small RNAs may play a significant role in mediating wound healing. The overarching hypothesis of this proposal is that oral-piRNAs are intrinsic factors in human oral keratinocytes, that modulate the rate of wound closure. Two specific aims are in place to further characterize the wound healing phenotype and elucidate the cellular factors responsible for oral-piRNA-mediated phenotypes. Aim 1 is to investigate the role of saliva piRNAs on oral wound re-epithelialization using 3D ex vivo and in vivo oral wound healing models. Aim 2 is to elucidate the mechanism by which piRNAs regulate oral wound healing. This proposal aims to unravel how these novel oral-piRNAs accelerate human oral keratinocyte proliferation and migration, which may improve wound healing therapeutic developments. The long-term goals of this proposed study are to enhance our understanding of the biological roles of piRNA relevant to human oral health and diseases and leverage these findings to improve wound healing outcomes for patients.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT The mammalian heart does not robustly regenerate after myocardial infarction and heals via a fibrotic repair response. Scar tissue is non-contractile, increases the hemodynamic burden on the remaining cardiac muscle and over time adverse cardiac remodeling occurs leading to ventricular dilatation and development of heart failure. Understanding mechanisms of ventricular remodeling to redirect the cardiac injury response from a fibrotic to a reparative one remains one of the broad therapeutic goals in cardiovascular medicine. We have recently identified extracellular nucleotide metabolism as a novel target for cardiac remodeling after ischemic cardiac injury. We demonstrated that after cardiac injury, the ectonucleotidase ENPP1 is upregulated and hydrolyzes extracellular ATP into AMP. Increased AMP was converted into adenine and extracellular adenine initiated a pro-inflammatory and pro-apoptotic cascade that caused cell death by disrupting NAD and pyrimidine biosynthesis in non-myocytes and myocytes. However the mechanisms of adenine generation and transport across the cell membrane, molecular and biochemical mechanisms of downstream effects on NAD and pyrimidine biosynthesis and whether the ENPP1/AMP/Adenine axis can be targeted for therapeutic gain to attenuate post infarct cardiac remodeling are unknown. In this proposal, we form a multi-disciplinary team comprising experts in cardiac physiology, metabolomics, computational biology, and finally population genetics to investigate the role of adenine metabolism in cardiac remodeling, identify its relevance as a potential therapeutic target and its association with human cardiomyopathy.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT CD8+ cytotoxic T lymphocytes (CTLs) play a key role protective role in HIV-1 infection, but cannot fully suppress the virus because they require antigen for maintenance. HIV-1-specific CTLs mediate the suppression of HIV-1 for the “asymptomatic” phase of chronic infection. The CTL response partially controls infection, but once the antigen is cleared to very low levels, they decay to low frequency resting central memory cells. Since CTLs require antigen to maintain effector function, they cannot fully suppress even in most “elite controllers,” who exhibit pathogenic low level infection. To suppress HIV-1 fully, CTLs need to be maintained independently of HIV-1 replication. Cytomegalovirus (CMV) drives CTL persistence through frequent low grade reactivations. Frequent spontaneous non-pathogenic reactivations of CMV drive persistent maintenance of CMV-specific active effector CTLs. We hypothesize that creating CTLs recognizing both CMV and HIV-1 can yield superior control of HIV-1 infection. Such CTLs would be maintained by natural CMV reactivations to be primed as active effector cells against HIV-1. To achieve this goal, we have developed a prototype bi-specific chimeric antigen receptor (CAR) that recognizes both CMV and HIV-1, coupling maintenance to CMV and de-coupling maintenance from HIV-1. This project will expand that effort; specifically we aim: 1. To optimize a panel of CMV and HIV-1 bi-specific CARs; 2. To evaluate the function of these CARs in the BLT humanized mouse model.

NIH Research Projects · FY 2025 · 2024-07

SUMMARY Pulmonary Arterial Hypertension (PAH) is a pulmonary vascular disease characterized by increased pulmonary arterial pressure. PAH patients are young, have poor life quality, and have a short life expectancy. This daunting fact underscores our need for innovative approaches in PAH. PAH arises from a pro-proliferative and anti-apoptotic phenotype of pulmonary arterial smooth muscle cells (PASMC), endothelial cells (PAEC), and fibroblast. Recent advances linked this phenotype to a genome-wide deregulation of gene expression. Therefore, it is imperative to decode the cause of this deregulation to find new therapies. Regulation of gene expression is governed, in large part, by one main class of protein: RNA-binding proteins (RBP). RBPs regulate the fate of hundreds of transcripts at once through the recognition of specific motifs that confer them a high therapeutic potential. However, limited data exist on their implication in PAH. Here, using large-scale transcriptomic data from PAH patients' lungs, and cultured PASMC and PAEC, we found ZFP36 an RBP increased in lungs and further enriched in PASMC but not in PAEC. We used computational biology to identify the targets of ZFP36 and characterize their functions in PAH-PASMC. Our data show that ZFP36 targets are lead actors in DNA repair and resistance to apoptosis (e.g. RFC3) and cell cycle (e.g. ANAPC4). In-vitro experiment on PAH-PASMC confirmed the up-regulation of ZFP36 targets and the role of ZFP36 in their up- regulation, as well as the role of ZFP36 in DNA repair and cell cycle. In addition, we found that inhibition of ZFP36 in Sugen/Hypoxia rats reduced PH severity.

NIH Research Projects · FY 2025 · 2024-07

Project Summary Systemic diseases such as diabetes mellitus and metabolic syndrome affect multiple organs of the body. While the human body is naturally capable of self-healing, it faces an increasing challenge as multiple components of the systems of the human body go awry. Metabolism is a dynamic network of biochemical reactions that support cell proliferation and biosynthesis. On the whole-body level, metabolic networks of individual tissues and organs are connected by the circulatory system and interfaced with the digestive and excretory systems. Our ability to cure systemic diseases relies on a quantitative understanding of whole-body metabolism, which requires comprehensive measurement of its dynamic states. However, challenges arise from the lack of our ability to quantify metabolic fluxes (i.e., rates at which pathways are utilized) on a systems level. Metabolic fluxes are a direct readout for the dynamic state of metabolism but intangible deduced quantities that result from the catalytic interaction between metabolites and enzymes according to the kinetic and thermodynamic laws. Metabolic flux analysis (MFA) framework allows quantitation of metabolic fluxes by imposing mass balances on all isotopologues resulting from stable isotope tracing experiments. As carbons form the molecular backbone, 13C-labeled substrates are extensively employed. The overarching aim of this project is to facilitate the measurement of metabolic fluxes on muti-tissue and whole-body levels by tracing multiple isotope tracers. Knowledge of metabolic fluxes offers dual benefits of laying a solid foundation for understanding and controlling metabolism. To effectively achieve this computationally intensive goal, our teams at UCLA and Stevens will combine deep learning with analytical, stable isotope tracing, and simulation techniques. Using multilayer neural networks, we will develop deep learning models that predict metabolic fluxes from the isotope labeling patterns of metabolites. With the augmented flux determination capability, we will impart quantitative systems-level knowledge of metabolism in individual and across tissues in co-cultures and animals.

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract Hispanics are one of the fastest growing populations of older adults in the United States (U.S.). From 2019 to 2060, the population of Hispanic older adults aged 65 years and over in the U.S. is projected to increase from 9% to 21%. Existing literature suggests that Hispanic older adults experience health disparities across multiple sectors of the U.S. healthcare system, including emergency medicine. Emergency medical services (EMS) provided in the prehospital setting, however, are a largely understudied sector in the U.S. healthcare system, especially in the context of Hispanic older adults. EMS serve as an important entry point into the U.S. healthcare system for Hispanic older adults who often have challenges accessing preventive and diagnostic care. The objectives of the proposed dissertation are three-fold: 1) assess the individual, neighborhood, and structural-level factors that impact the provision of EMS care by emergency medical technicians and paramedics to Hispanic older adults; 2) examine the individual and neighborhood-level factors that impact the provision of EMS to Hispanic older adults experiencing a high-acuity, time sensitive, cardiac-related 9-1-1 emergency; and 3) determine the individual and neighborhood-level factors that impact the provision of EMS for Hispanic older adults experiencing a psychiatric-related 9-1-1 emergency, a type of call often triaged as low acuity. The National Institute of Aging’s Health Disparities Framework provides the theoretical foundation for the three studies that comprise this dissertation. The first study in this dissertation is a systematic literature review, an increasingly important approach to synthesize existing literature and provide justification for future research. The second and third studies use data from three sources, including the San Francisco Department of Emergency Management, San Francisco Department of Public Health, and the 2020 and 2021 American Community Survey. The PRISMA 2020 guidelines and Covidence software inform the completion of the systematic literature review, which addresses the first objective. Logistic regression and multilevel modeling are used to: 1) examine the association between individual and neighborhood-level factors and high- acuity, cardiac-related 9-1-1 emergency calls, which address the second objective; and 2) assess the association between individual and neighborhood-level factors with low-acuity, psychiatric-related 9-1-1 emergency calls, which address the third objective. Neighborhood in the second and third objectives refers to ZIP code as the level of geospatial analysis. Stata 17.0 and R are used to run the statistical analyses in the second and third studies. The proposed dissertation will offer insights on the provision of EMS to Hispanic older adults. The findings from this dissertation can inform ongoing efforts to make the EMS system more accessible, age-friendly, and equitable.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Cystic fibrosis (CF) is a chronic, multi-organ process characterized by inspissated secretions that adversely impact the upper and lower airway, gastrointestinal tract, and other organ systems. Chronic rhinosinusitis (CRS) is a prevalent and clinically impactful disease complication in individuals with CF. CRS is detrimental to quality of life, impairs pulmonary status and worsens overall disease state. Olfactory dysfunction is a key feature of CRS, further worsens quality of life, and is associated with depression and nutritional alterations. These comorbidities are significantly understudied in young children with CF (YCwCF). Highly effective CF transmembrane conductance regulator modulator therapy (HEMT) improves pulmonary health and certain extra- pulmonary domains for adults with CF. Previous research showed that adults with CF have substantial improvement in CRS after HEMT initiation. However, prior work demonstrated that adults treated with HEMT did not have improvements in olfactory dysfunction or have complete resolution of sinus disease. Given that airway inflammation and infection are present for a short time in young children (before starting HEMT), we anticipate that initiating therapy at an early age will lead to substantial improvement in CRS and olfactory dysfunction. The goals of this study are to characterize CRS and olfactory dysfunction in YCwCF and test the hypothesis that these comorbidities improve with HEMT. This prospective, observational, multi-center study will pair with a prospective, observational, multi-center study of outcomes in YCwCF that will investigate changes in other domains (pulmonary, microbiological, endocrine, and gastrointestinal) following initiation of HEMT. In this study, CRS and olfactory dysfunction will be evaluated using validated, age-appropriate, objective and subjective, non- invasive outcome measures. A treatment group will be comprised of YCwCF who initiate HEMT. A control group will be comprised of YCwCF who do not initiate HEMT. Both groups will be followed for two years and the groups will be age-matched. Analysis will use mixed-effect models for longitudinal data to compare outcomes between groups and incorporate propensity scoring. Specific Aim 1 will characterize CRS and olfactory dysfunction in YCwCF. Specific Aim 2 will test the hypothesis that HEMT improves CRS in YCwCF utilizing sinus magnetic resonance imaging opacification and sinonasal symptom burden. Specific Aim 3 will test the hypothesis that HEMT improves olfactory dysfunction in YCwCF using olfactory bulb volume, quantitative olfactory function, olfactory cleft opacification, and olfactory-specific quality of life. Findings from this research will inform strategies that will lead to greater overall health status and improved quality of life. Results will highlight the importance of complication evaluation and management early in life.

NSF Awards · FY 2024 · 2024-07

This project aims to advance mathematical theories of phase transitions and critical phenomena in a number of specific contexts. While these effects are ubiquitous in physics and chemistry, understanding them mathematically is challenging due to the need to consider the limit of large systems whose constituents exhibit persistent dependence throughout all scales. Probability theory offers a number of tools and proof strategies for this purpose provided the problem is cast, or can otherwise be represented, via elementary probabilistic concepts such as random walks or random fields. The goal of the analysis is to show that, in several settings, as the system size increases, a new structure called the scaling limit appears. Often quantitative details of the scaling limit are often independent of the specifics of the original problem, demonstrating universality. . Through the specific examples to be studied, new approaches and techniques will be developed that will advance our understanding of scaling limits and phase transitions. The questions are of varied difficulty, which will make it possible to include graduate students and postdocs in this research. The project explores a diverse list of specific questions from probability, mathematical physics and analysis. First, extremal processes of branching Markov chains, local time of random walks and other random fields exhibiting logarithmic correlations will be studied with the aim to establish broad universality of the Gaussian Free Field and develop novel techniques for dealing with these problems. Second, scaling limits of random walks in random environments will be analyzed in examples that test the limits of the existing theory of stochastic homogenization. Here the particular emphasis is on the situations where the underlying environment has good spatial mixing properties and yet is not necessarily drawn from a stationary probability law. Third, a recent treatment of Bose-Einstein condensation in mean-field interacting bosonic gases using Feynman’s random-cycle representation will be expanded to establish a quantum phase transition. The point is to provide a direct link between the properties of the quantum system and those of its graphical representation. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-07

Project Summary/Abstract Dr. Hsue is one of the few cardiologists in the world with both extensive research and clinical expertise in HIV and has devoted the last 15 years studying cardiovascular disease (CVD) in the setting of HIV. The initial part of her career was spent 1) establishing a HIV Cardiology Clinic at Zuckerberg San Francisco General (ZSFG), 2) developing the infrastructure to perform patient oriented research in Cardiology including development of a vascular laboratory for clinical research, 3) performing studies that evaluate the mechanisms underlying HIV infection and CVD and 4); leading clinical trials aimed to reducing inflammation and CVD risk in HIV. The findings from her work have shown that 1) HIV infection is independently associated with CVD, independent of traditional risk factors or ART and 2) chronic inflammation in the setting of effectively treated HIV infection underlies this increased CV risk. Initial K24 support has led to a significant increase in the number of mentees, grants, and publications for Dr. Hsue and new research directions including use of novel imaging to assess HIV disease burden and identification of therapies to reduce HIV-associated inflammation and CV risk. For the K24 renewal period, her career goals are to perform cutting edge clinical/translational studies and mentoring that will lead to improvements in treating, identifying and preventing CVD among PLWH and also impact other comorbidities and HIV cure. Dr. Hsue plans to 1) expand her research to include the role of somatic mutations in the hematopoietic system (termed clonal hematopoiesis of indeterminate potential, CHIP) and proteomics in HIV-associated CVD; 2) continue her time spent mentoring junior investigators in the fields of HIV infection, inflammation, and CVD with a focus on transition to full-independence; 3) obtain leadership training to build clinical and translational research programs that can be used by mentees and will make her a better mentor. The majority of Dr. Hsue's career development will occur at UCSF which offers outstanding resources for clinical/translational research, basic science, imaging, HIV, and mentoring/leadership training. Her K24 proposal will extend upon her current NIH-funded investigations by studying the underlying mechanism of HIV- associated atherosclerosis in treated HIV with the following Specific Aims: Aim 1A: To determine if clonal hematopoiesis of indeterminate potential (CHIP) mutations are more prevalent in peripheral blood cells of PLWH vs. uninfected controls; Aim 1B: To determine whether the presence of CHIP mutations are associated with increased arterial inflammation and metabolic activity of the hematopoietic system as assessed by FDG- PET/CT; Aim 2A: To identify a unique proteomic signature in PLWH that is associated with arterial inflammation and metabolic activity of the hematopoietic system, and Aim 2B: to characterize changes in proteins and biological pathways that are altered after anti-inflammatory and lipid interventions. This proposal will provide subject visits, imaging, and clinical and laboratory data to support new investigators in clinical/ translational research in the field of HIV-related CVD and HIV disease pathogenesis including cure.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY The goal of this proposal is to investigate how gene expression dosage on the X chromosomes is regulated during embryonic and extraembryonic development and how its deregulation impairs the feto-maternal interface within the placenta. X-chromosome dosage compensation in female placental mammals occurs through two epigenetic processes: X-chromosome inactivation (XCI), which induces the inactive X chromosome (Xi), and X- chromosome upregulation (XCU), which increases the expression of the single active X chromosome (Xa) to match the gene dosage of autosomes. XCI happens twice in mouse development. First, an imprinted mechanism silences the paternal X (imprinted XCI, iXCI) in pre-implantation embryos. iXCI is reversed in epiblast cells of the blastocyst and induced again on either the maternal or paternal X in embryonic cells upon implantation (random XCI, rXCI); in contrast, extraembryonic cells, which form the placenta, maintain iXCI. XCU accompanies both iXCI and rXCI. XCI mechanisms are well-understood, but those governing XCU remain largely elusive. The long non-coding RNA Xist is the master regulator of both forms of XCI. Conserved repeats of Xist, termed A-F, are critical for Xist’s function and recruit specific proteins to block transcription and induce a heterochromatin state. For instance, the B-repeat recruits repressive proteins including polycomb complexes and is required for heterochromatin formation and Xi compaction. We showed that without the B-repeat, silencing of most genes on the Xi is impaired in vitro. To investigate the functional consequences of increased gene expression from the Xi, we created mice with a B-repeat deletion (ΔB-Xist). Our preliminary data show that the ΔB-Xi disrupts the development of the placenta of female embryos. Female ∆B-Xi placentas have severe depletion of the junctional zone (JZ) and limited trophoblast invasion of blood vessels, causing female-specific embryo mortality and growth retardation. In the ΔB-Xist mouse model, developmental defects only manifest in the placenta, even though gene expression on the ΔB-Xi is increased in placental and embryonic tissues. Our data suggest that Xa expression is typically decreased in cells with the ΔB-Xi and that this does not happen in the JZ, suggesting that XCU regulation varies in different cell types and providing a possible explanation for the JZ-specific sensitivity to X- chromosome dosage. Here, we will follow up on these exciting findings by further defining the role of the Xist B- repeat in the placenta (Aim1); exploring the chromatin state on the wildtype and ΔB-Xi and the crosstalk between the Xi and Xa in embryonic and extraembryonic tissues (Aim 2); and uncovering mechanisms underlying JZ dysplasia in placentas of mouse interspecies hybrids (Aim 3). Overall, our work will elucidate the role of XCI and XCU in the regulation of the feto-maternal interface and explain the high X-dosage sensitivity of the JZ.

NSF Awards · FY 2024 · 2024-07

The magnetosphere is the region that surrounds the Earth, which is carved out by its magnetic field as it deflects the supersonic solar wind plasma around it. The solar wind is a major energy source for the magnetosphere that can lead to complex dynamics. One such dynamic is the magnetospheric substorm, a major system reconfiguration that results in an energy release in Earth's magnetosphere-ionosphere system. During a substorm, a large-scale current system is observed known as the substorm current wedge (SCW). This project's focus is to better understand the physical process that leads to the SCW. Another example of the complex behavior observed in the magnetosphere is the presence of sporadic fast flows, known as bursty bulk flows (BBFs). This project seeks to investigate if there is any connection between the formation of the SCW and BBFs. The project's successful outcome will lead to a better understanding of the Earth's magnetosphere and current systems. This project will support two early-career scientists. Results of the study will be conveyed to the public (including K-12 students) via education and public outreach eWorts in both the PI and Co-I's institutes, which will enhance public interest in space science. The research focuses on the substorm current wedge (SCW), which plays a crucial role in the energy release process within Earth's magnetosphere-ionosphere system. This research aims to explore the causes behind this asymmetry and investigate the validity of the collective wedgelet formation of a SCW. Specifically, the team suggested to answer the following questions: Q1. Do the thermal pressure asymmetries around DFBs result from an interplay between the meso and global scales? Q2. Which plasma populations contributed to the pressure asymmetry within the dipolarization front layer? Q3. Are the collective eWects of wedgelets consistent with a substorm current wedge? The team will use the Multiscale Atmosphere-Geospace Environment (MAGE) model, inertialized Rice Convection Model (RCM-I) numerical simulation, and THEMIS observations to achieve the science goals. The outcome of our studies will provide constructive information on the physics of the inner magnetosphere dipolarization process and advance our understanding of the nature of substorm current wedge formation. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT Globally, populations are aging thereby increasing healthcare burden, overall cognitive impairment, and dementia including Alzheimer's disease (AD). The lack of effective treatments makes it essential to develop new strategies for healthy cognitive aging, including interventions to slow or prevent cognitive decline. A traditional Mediterranean diet, rich in polyphenols (PPs), may prevent or delay the onset of cognitive dysfunction in older adults, preserving healthy brain structure and function, and lowering the risk of AD. These effects, mediated in part by gut microbiome-derived PP metabolites, highlight the role alterations in the brain-gut microbiome system play in neurodegeneration. Moreover, high levels of circulating phenyl-γ-valerolactones, neuroprotective compounds, exclusively produced by gut microbiota from flavan-3-ol-rich foods (e.g., cocoa, tea, berries) are associated with delaying the onset of cognitive dysfunction in older adults. Intake of such PPs can also change gut microbial composition and function, altering the physiology of the host's secondary bile acid (BA) pool, affecting regulatory and signaling functions in the brain as well as cognitive decline and AD. We hypothesize that, in older adults with enhanced AD risk, dietary intake of PPs maintains healthier brain features and cognitive function, and that this beneficial effect is mediated by gut microbiota metabolites of PPs and BAs. In this multi- PI application by leaders in the field of brain-gut microbiome interactions, we will conduct a year-long, multi-center randomized double-blind placebo-controlled study in 300 older adults in the United States (validation sample of 100 from Northern Ireland) who are at enhanced risk of developing AD. Coupled with this, behavioral interactions will be investigated with a murine fecal microbiome transplant model incorporating samples from high AD risk participants on a high PP supplement compared to a placebo. We will apply multiple complementary approaches: meta and transcriptomic interrogation of the gut microbiome, targeted/untargeted metabolomics for microbiota- derived metabolites, multimodal brain imaging, assessment of cognitive function and inflammatory status, and advanced bioinformatic techniques for data integration. The hypothesis is addressed as follows: Aim A: Identify the protective effects of high intake of supplementary dietary PPs on brain and cognitive parameters in high AD- risk participants. Aim B: Determine the effect of PP intake on the microbiome, inflammatory, and AD biomarkers in high AD risk participants. Aim C: Explore causal relationships between PP intake and gut microbial metabolites, inflammatory and AD markers, brain parameters, and cognitive function. Aim D: Utilize a reverse translational approach to identify changes in mouse brain and behavior by PP-induced alterations in the human gut microbiome. Ultimately, we will establish the protective effects of regular dietary PP intake on cognitive function and on brain-gut microbiome interactions, ideally allowing the development of effective dietary regimes to prevent or delay the onset of AD in at-risk elderly, thereby reducing cognitive decline and healthcare costs.

NIH Research Projects · FY 2024 · 2024-07

Project Summary The goal of this project is to engineer a g-aminobutyric acid (GABA) oxidase suitable for the creation of a high- performance, implantable, electroenzymatic GABA microsensor. GABA is the most important inhibitory neurotransmitter in the brain, yet technology to monitor GABA transients at the cellular level and at the near- real-time, millisecond time-scale has been slow to emerge, especially for the deep brain. The need is acute, as glutamate and GABA dominate the excitatory/inhibitory (E/I) ratio in the brain, dysregulation of which has been associated with Alzheimer’s disease, seizures, autism, schizophrenia and bipolar disorder, which affect nearly 25M people in the US. Since GABA itself is not electroactive, a proven approach to creation of a high- performance sensor entails the immobilization on an electrode of a H2O2-producing oxidase selective for the analyte of interest. The H2O2 produced by the enzyme-catalyzed oxidation of the analyte is electrooxidized at the underlying electrode thereby giving rise to a current that may be correlated to the analyte concentration. However, a GABA oxidase is not commercially available, and other biosensing strategies have significant drawbacks. Since GABA is ubiquitous in the environment, there is evidence for enzyme activity closely related to that necessary for sensor development. In fact, an enzyme with native oxidase activity for closely related methyl-GABA that shows modest activity with GABA has been identified. Our team has cloned and expressed this enzyme, demonstrated its activity with GABA and constructed a biosensor showing modest ability to sense the target. Our aims are to employ directed protein evolution to create a highly active and selective GABA oxidase using advanced ultrahigh throughput techniques and to demonstrate its utility for the construction of a high-performance GABA microbiosensor. Promising prototype biosensors will be tested as they are created in the striatum of anesthetized rats. This project will leverage the proven expertise of the team in the areas of protein engineering; of neuroscience; and of implantable, electroenzymatic microsensor construction to create technology that will help further illuminate the function of exceedingly important neural circuitry.

NIH Research Projects · FY 2026 · 2024-07

Abstract Most symptomatic Latina women do not receive a diagnosis or initiate mental health (MH) treatment (Tx) when warranted. Stigma, cultural concerns, distrust of professionals, lack of insurance, and not knowing where to get help are barriers so many suffer from untreated depression and/or anxiety. Due to high social media and YouTube use and high demand for story-based media among Latinas, however, our multidisciplinary team created and tested a choice-driven, Hollywood-quality web-based app featuring transmedia storytelling videos called Catalina: Confronting My Emotions/ Enfrentando Mis Emociones (in English and Spanish) to help Latinas overcome barriers to initiate MH Tx. Multiple waves of theater-testing and pilot testing showed symptomatic, untreated Latinas reported high identification with the Latina lead-character and high trust of the Latina nurse-therapist character; all was developed using composite data from past qualitative studies with Latinas. Using transmedia (storytelling across multiple digital platforms), the story expands beyond a traditional episode through character-driven, interactive bonus videos and social media posts of the characters that are also psychoeducational plus a character’s blog with links to free MH resources and recommendations of local clinics offering low-cost Tx plus Tx locator. A Latinx cast, director, and script writer enhance socio-cultural acceptability and bring Latinas back to re-watch videos. Access is discreet via smartphone, tablet, or computer. Guided by Ajzen’s Theory of Planned Behavior and supported in feasibility and efficacy testing, we identified targets that mediate help-seeking behavior change including attitudes, propensity to seek help, and stigma. For this 2-arm RCT, Aim 1 is to determine the effectiveness of the Catalina experimental transmedia web-based app compared to a control condition (informational MH videos but no story, links to publicly available MH websites, and Tx locator for low-cost options) across 9 months (baseline, 1, 5, and 9 months) to catalyze therapy or pharmacotherapy initiation. Both groups will receive monthly reminder messages via text or email. Consent procedures allow for verification from medical records if MH Tx appointments were made, kept, or missed and if prescriptions were filled. We hypothesize that, compared to controls, a significantly greater proportion of the Catalina group will initiate MH Tx during the 9-month study. Aim 2 is to test the mediation role of the targets (help-seeking attitudes, propensity to seek help, and stigma) across 9 months to initiate MH Tx in Catalina vs. control groups. The sample will include 876 English and Spanish speaking Latinas, age 18 and older, who are untreated but symptomatic for depression and/or anxiety (>10 on PHQ8 or GAD7) and who have internet access. All aspects of the study are done online. If hypothesized relationships are supported this study has potential for high impact on English and Spanish speaking Latinas, for whom viable MH solutions are often less accessible. Knowledge gained about drivers that catalyze Tx initiation will expand strategies to help symptomatic Latinas overcome barriers and connect to needed MH Tx via our scalable app for low cost.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Cell division is a complex process that is tightly regulated by the activation and inactivation of a series of proteins that ensure the proper segregation of genetic material into two daughter cells. Misregulation of key mitotic events, including chromosome alignment and segregation, can lead to genetic mutation that promotes abnormal cell proliferation and tumorigenesis. To better understand cell division and its misregulation in human disease, we performed a genetic RNAi screen for novel mitotic regulators and identified the phosphatase DUSP12. While DUSP12 has been shown to possess pro-survival attributes, it is unclear if this is related to its potential role in cell division. Preliminary proteomic analyses of DUSP12 using affinity- and proximity-based approaches coupled to mass spectrometry identified ZPR9 as a potential interactor of DUSP12 which has been validated by in-cell and in-vitro IP assays. Previous studies have shown that ZPR9 may play a role in apoptosis, but it is unclear if and how ZPR9 participates in cell division and, more so, how it promotes cell death. The central hypothesis of this project is that DUSP12 is a novel mitotic regulator and it influences cell fate by de-phosphorylating its target substrates, including ZPR9, which plays a critical role in the processes of cell death and proliferation. Aim 1 will characterize how DUSP12 modulation influences mitotic progression. Using cancer cell lines as a model system and leveraging advanced imaging techniques, we will explore the temporal and spatial activity of DUSP12 and its impact on key mitotic events and machinery. Aim 2 will evaluate the molecular mechanisms through which DUSP12 influences cell death and survival and if and how this applies to its role in cell division. These studies will bring new insights to our understanding of the importance of protein phosphatases for cell division, advancing our understanding of the full complement of proteins required for mitotic regulation and their relevance to human health and disease. Collectively, this work will provide insights into the mechanism and regulatory role of the phosphatase DUSP12 during cell division as well as form the basis for designing novel therapeutic approaches to address diseases associated with abnormal proliferation.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Ongoing evaluation of environmental cues that predict aversive outcomes enable animals to avoid threats. However, learned avoidance can easily become maladaptive. Excessive or inappropriate avoidance is a core feature of numerous psychiatric disorders (e.g., depression, anxiety, PTSD and OCD). Although pathological behavior in these conditions is linked to dysfunction in the prefrontal cortex, we lack the detailed neurobiological understanding necessary to design precisely targeted therapeutic interventions. The medial prefrontal cortex (mPFC) is required for learning associations that drive approach or avoidance behaviors, but the circuit mechanisms that underlie this process are poorly understood. Prefrontal dopamine (DA), in contrast with subcortical DA, most strongly encodes aversive stimuli. DA has a well-established role in plasticity and shapes activity in projection-specific mPFC neuronal populations expressing either D1 or D2 receptors (D1R or D2R). However, the role of prefrontal DA, and D1R+ vs. D2R+ populations, in avoidance learning remains largely mysterious. We will address these deficits using innovative tools that allow us to measure and/or manipulate DA and mPFC neuron populations while mice learn to avoid threats. We will test the specific hypothesis that DA generates an mPFC activity state required for flexibly learning to associate cues with actions that preempt aversive outcomes. In Aim 1, we will use temporally precise measurement and manipulation of prefrontal DA to thoroughly interrogate the timing and causality of DA dynamics during threat avoidance learning. In Aim 2, we will use miniaturized head-mounted microscopes and simultaneous optogenetic silencing of mPFC-projecting DA neurons to determine the causal role of DA in modifying emergent neural activity patterns required for avoidance learning. Finally, in Aim 3, we will focus on D1R+ and D2R+ cell-type specific circuits by recording their activity and electrically silencing them during threat avoidance learning. This proposal directly addresses a pressing need to understand the cell-type and circuit-specific mechanisms that mediate avoidance learning. Our research can inform pharmacological, psychotherapeutic and brain stimulation interventions for a variety of psychiatric conditions characterized by maladaptive avoidance.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT New SARS-CoV-2 variants and novel coronaviruses will constantly arise and affect large proportions of the worldwide population. Although the current wave of infections with Omicron subvariants is much less lethal than that caused by previous SARS-CoV-2 variants (e.g., Wuhan-Hu1, Alpha, and Delta), about 10% of Omicron- infected patients develop prolonged post-acute neurological sequelae involving cognitive impairments (“brain fog”), difficulty concentrating, anxiety, and depression. Histological studies of brains from COVID-19 patients have often observed immune cell infiltrates and increased frequencies of glial cells with inflammatory phenotypes indicative of neuroinflammatory responses. Currently, there are no approved prophylactic or interventive treatments for SARS-CoV-2-induced neuroinflammation. There is a growing body of evidence that immune cells, microglia, and astrocytes express γ -aminobutyric acid receptors (GABAA-Rs). The activation of these GABAA-Rs inhibits their pro-inflammatory activities and shifts T cells, antigen-presenting cells, microglia, and astrocytes toward anti-inflammatory phenotypes. Taking advantage of these properties, researchers have used homotaurine, a clinically applicable GABAA-R agonist that can cross the blood-brain-barrier, to 1) reverse experimental autoimmune encephalomyelitis (EAE) a model of multiple sclerosis (MS) and 2) reduce the levels inflammatory cytokines in the CNS, decrease neuronal loss, and ameliorate behavioral deficits in a model of autism spectrum disorder. Furthermore, we have shown that GABAA- R agonists reduce circulating proinflammatory cytokines/chemokines, viral replication in the lungs, the severity of illness, and death in mice infected with two different coronaviruses: MHV-1 and SARS-CoV-2. Additionally, in clinical trials, homotaurine treatment has had some promising beneficial effects in Alzheimer’s disease patients. We contend that homotaurine is an excellent candidate for preventing and ameliorating SARS-CoV-2- induced neuroinflammation. We will test this hypothesis in the hamster model of long COVID, arguably the best rodent model available at this time. We anticipate that we will demonstrate the potential of an entirely new drug class to help prevent and ameliorate SARS-CoV-2-induced neuroinflammation. Because homotaurine is safe for human consumption, it could be rapidly tested in clinical trials. Since the mechanism evoked by GABAA-R activation is unlike that of any other drugs being considered for treating long COVID, GABAA-R agonists are likely to have enhanced therapeutic effects when combined with other treatments. Finally, the results may have broader applications to treat other conditions involving CNS inflammation. Therefore, the results of our studies may provide novel approaches to help improve the health of the global community.