University Of California, San Diego

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Hispanics/Latinos are at increased risk for developing Alzheimer’s disease and related dementias (ADRD) compared to non-Hispanic Whites. While factors underlying this disparity are not well understood, increased cardiovascular disease (CVD) risk among Hispanics/Latinos is likely to be a contributory factor. Pathological changes of ADRD begin years before clinical symptoms become evident and interventions are most likely to confer benefit in the earliest stages of ADRD. There is a pressing need to develop tools to detect the earliest manifestations of ADRD, particularly in Hispanics, who develop symptoms of ADRD at earlier ages, yet are diagnosed at more advanced disease stages than other groups. A variety of behaviors show changes in the preclinical stages of ADRD, including sleep, gait speed, and physical activity, among others. Recent innovations in mobile technology now offer novel ways to collect, track, and analyze these behaviors passively and unobtrusively, as a person engages in their daily life. Our preliminary work demonstrated that the application of machine learning models to passively-collected digital health data from smartphones and wearables differentiated persons with and without mild cognitive impairment with 85% accuracy in a primarily non-Hispanic White sample. Guided by the NIA Health Disparities Research Framework, we propose to leverage artificial intelligence (AI)-powered analytics and insights, coupled with readily available sensors in consumer electronics (smartphones, wrist-worn wearables), to identify digital biomarkers of ADRD risk, with a focus on vascular contributions to dementia, among Hispanics/Latinos. Augmenting an existing cohort study of Hispanics/Latinos residing in Southern California, the proposed study has three principal aims: Aim 1 involves determining digital signatures of ADRD risk among Hispanics (N=300; aged 50-70 years) using integrated passive mobile sensing features, derived from smartphones and wrist-worn wearables, and machine learning methods. ADRD risk will be defined by cognitive status and CVD risk burden (diabetes, hypercholesterolemia, hypertension, obesity, smoking) and will also incorporate apolipoprotein E (APOE-ε4) and plasma-based AD biomarkers for further classification of ADRD risk. Aim 2 investigates sex differences in digital signatures of ADRD risk and Aim 3 examines the impact of sociocultural factors (e.g., language use, acculturation) on these signatures. We will also investigate whether changes in digital data features predict longitudinal neurocognitive change over a span of three years in a subset of Hispanics with and without ADRD risk. Housed within a renowned research institution at the vanguard of ADRD research and engineering innovations, the proposed study includes a multidisciplinary team with expertise across all aspects of this cutting-edge proposal. Recognizing the value of a community- engaged research approach, we have partnered with community stakeholders to ensure the relevance of our study to the Hispanic community. Our work could revolutionize early detection of ADRD and reduce ADRD disparities by developing a low burden, low-cost approach to identify ADRD risk among Hispanics.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Substance use disorders are among the most common psychiatric disorders and are a leading cause of disability throughout the world. Over the last decade, genome-wide association studies (GWAS) have identified numerous genetic loci that contribute to addiction (opioid, tobacco, cannabis, alcohol). Turning these discoveries into mechanistic insights, a necessary first step for understanding the pathophysiology of addiction, and ultimately leading to the development of new therapies, is challenging. Model organisms serve as excellent tools to understand how human genomic variation affects traits. However, integration of human GWAS data with studies in model organisms has been limited. This is because GWAS do not lead to the identification of genes, but genetic variants (or SNPs), which cannot be easily translated across species. In addition, human GWAS have shown that risk for addiction is highly polygenic, but the existing strategies for cross-species translation do not capture the polygenic architecture of addiction. I am proposing an innovative solution to this problem by developing a framework for cross-species polygenic translation. Polygenic risk scores (PRS), which are a widely used tool for human genetic studies, predict risk for a trait by summing the contributions of numerous SNPs. Because these SNPs are species-specific, it is not possible to apply a PRS to another species. I am proposing to use transcriptomic analyses to overcome this obstacle. Leveraging resources and techniques from well-established statistical genetic tools, I will develop a method that will allow translation of polygenic signals from humans to rodents, and vice- versa. This will be accomplished by using the following steps: 1) use GWAS for addiction-related traits in human and model organisms to compile catalogs of genetic variants; 2) use transcriptomic data from GTEx and analogous datasets from model organisms to build gene prediction models, allowing estimation of transcript levels in individuals based on genotype information, 3) determine the association between these estimated transcript levels and addiction-related traits and 4) use these gene-level associations (rather than SNP level associations) to calculate Polygenic Transcriptomic Risk Scores (PTRS). This approach translates SNPs into gene estimated gene expression levels and then takes advantage of the fact that, while the same SNPs do not exist across species, gene orthology can be used to translate between species. In addition, I will use complementary methodologies, including cross-species network analyses and other tools, that also account for the polygenic nature of addiction. Developing tools that allow polygenic studies to share information between humans and model organisms will be transformative by opening up entirely new lines of research. For example, PTRS will provide a novel means of validating animal models of addiction, as it will be possible to empirically test whether the genetic signature for addiction in humans is related to the genetic signature of addiction phenotypes in rodents. PTRS will also serve as a toolkit for drug repositioning, namely studies aimed at identifying small molecules and other interventions that can alter the global gene expression in model organisms in a way that lower risk, as predicted by PTRS and network- based analyses.

NIH Research Projects · FY 2026 · 2021-09

Abstract Three years ago, we proposed to apply our novel lipid chemistry platform to create highly potent, orally bioavailable compounds directed at SARS CoV2. By conjugating remdesivir monophosphate to lipophilic hydrocarbon side chains, we have developed a series of compounds that exhibit potent anti-SARS CoV2 activity. These compounds are well tolerated and highly bioavailable in mice and produce sustained, high-level intracellular exposure to remdesivir triphosphate. In proof-of-concept studies we demonstrated once-daily oral dosing of our compounds was superior to obeldesivir in a murine SARS CoV2 model infection. Because of their potency, favorable tolerability and pharmacokinetic properties, we evaluated their antiviral activity against several additional emerging viral pathogens. We have demonstrated antiviral activity at orally achievable levels of drug against a diverse set of pathogens with pandemic potential including Ebola, Marburg, Nipah, Hendra, Yellow Fever and Dengue viruses. Although we are not proposing to study these non-pandemic viruses in this application, further supporting their broad spectrum, our compounds were also highly active against measles and mumps virus. Recent studies extended the proof-of-concept to dengue virus by demonstrating full protection of ADE-enhanced immunodeficient mice from a lethal dengue virus challenge. In this renewal application, we propose to determine whether we can further enhance oral bioavailability of the compound series by utilizing formulations that favor micellar formation of prodrug in the gut. We wish to further explore structure-function relationships to determine whether we can create compounds with more potent antiviral activity and/or enhanced pharmacological properties. Finally, with the optimized compounds, we wish to conduct validation studies against additional pathogens of pandemic potential. We believe that these compounds have the potential to be efficacious when delivered orally once daily against several viral pathogens of medical importance including dengue and respiratory syncytial virus and that they could prove to be critically useful in early use in early use against several pathogens with significant pandemic potential.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Pregnancy relies on finely-tuned adaptations of the maternal immune system to establish and maintain immune-tolerance to the semi-allogeneic fetus, which appear to be imbalanced in women experiencing complications, such as preterm birth. Outside the pregnancy context, the central nervous system can modulate systemic immunity via the sympathetic nervous system, while its role in modulating maternal immune system adaptations during pregnancy remains unknown. Understanding the determinants of immune regulation throughout pregnancy is a prerequisite to advance the early risk assessment and treatment of pathological pregnancies. This research project proposes to target the interaction between the brain, pregnancy, and the immune system. State-of-the-art immunology (mass cytometry) and neuroscience (neuromodulation of discrete neuronal circuitry) approaches will be combined to comprehensively investigate neuroimmune regulation of human term and preterm pregnancies and mouse models of pregnancy. Specifically, we hypothesize that a hyperactive sympathetic (adrenergic) tone enhances systemic pro-inflammatory monocyte responses, thereby contributing to a compromised fetal tolerance in preterm pregnancies. The first aim of this proposal will monitor the responsiveness innate pro-inflammatory monocytes to adrenergic signals (using single-cell high-dimensional mass cytometry), and the nervous-system associated proteome (e.g., adrenergic analytes, Brice Gaudilliere lab) throughout the human third trimester. This aim is based on my current expertise in human pregnancy immunology and multi-parameter approaches, and a continuation of my prior in-depth analyses of the pre-labor plasma signature and immunome in the last 100 days of healthy pregnancy, which accurately predict the onset of labor. In aim 2, to understand brain-pregnancy communication on a mechanistic level, neuroscience approaches (fiber photometry and chemogenetics) will be used under supervision of the Luis de Lecea lab, who pioneered cutting-edge neuro-modulatory techniques. Here, I will manipulate discrete neural populations, known to either suppress or enhance peripheral immunity, throughout late gestation in mouse pregnancies with the aim to monitor their link to peripheral monocyte responsiveness. Reciprocally, fiber photometry will track pregnancy- specific alterations in activity of these neural populations. In the third and final aim 3, I will determine whether monocyte responses to sympathetic signals differ throughout preterm vs. term pregnancies by analyzing maternal blood collected in each trimester of pregnancy, using high-dimensional single-cell mass cytometry. Machine learning approaches will be utilized to train a model for preterm vs. term sample classification. In the independent R00 phase, I envision to apply techniques from immunology and neuroscience to understand pregnancy biology and pathology from a central nervous system perspective. Together, the set of proposed studies will answer fundamental questions in the nascent field of brain- peripheral immunity cross-talk, and elucidate understudied mechanism of neuroimmunomodulation in pregnancy, while significantly aiding in my career development and academic transition to independence.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT The goal of this project is to significantly advance the field of acute and semichronic epilepsy monitoring using novel, high-resolution electrocorticography (ECoG) record/stimulate grids (4096/256 channels, respectively) and stereoelectroencephalography (sEEG) depth electrodes (120/8 micro/macro) with full wireless data and power transfer. This project builds on our previous success in conducting the first-ever human trials for acute mapping of eloquent brain tissue with multi-thousand channel microelectrode grids. The proposed system encompasses multiple transformative technological approaches, including: (1) leveraging advanced thin-film microfabrication on 8” diameter substrates, thus permitting long integrated connectorization from thousands of channels; (2) exploiting a newly developed platinum nanorod (PtNR) microelectrode technology with excellent low impedance, high charge-injection-capacity (4.4mC/cm2), stability, and biocompatibility; and (3) using a thin (~10μm) parylene C substrate that is compliant to brain movements, conformal to brain curvature, and transparent, permitting easier visualization of brain anatomy during the acute mapping. Further, (4) the grids developed for this project are modular and can be trimmed to fit different sizes of craniotomies, and (5) this system offers a new generation of minimally invasive sEEG electrodes with easily reconfigurable microcontact distribution in different regions of the brain Our proposed system also (6) employs state-of-the-art acquisition electronics with a miniaturized 1024ch neural interface system-on-chip and radio transmission of data and power, enabling fully wireless monitoring that eliminates wire externalization, and (7) deploys multi-screen and multi- window visualization of the whole repertoire of electrophysiological activity, with the option to display and interpret signals in standard fashion. Our goal is to demonstrate in the semichronic clinical setting a high-definition display of traditional and emerging clinical biomarkers for epilepsy monitoring and treatment. To achieve this goal, we will pursue in Aim 1 regulatory input from the FDA and scale our grids under good quality laboratory practices (GLP), and perform benchtop testing and hardware and software development under a quality management system. In Aim 2, we will perform semichronic animal testing under GLP to demonstrate safety, tolerability, and efficacy of the new epilepsy-monitoring system. In Aim 3, we will will perform pre-clinical and human intraoperative recordings with appropriate IRB authorization. We will pursue FDA clearance for semichronic implants in Aim 4, and transition Aim 5 to semichronic epilepsy monitoring in patients with intractable epilepsy. The methods employed in device and system development, surgical approaches, electrophysiology, and data analysis will not only advance functional and epilepsy monitoring but will also have significant implications for numerous applications in neuromodulation/therapeutic stimulation, minimally destructive brain-machine interfaces, and spinal cord stimulation.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY/ABSTRACT The blood-brain barrier (BBB) is a term that describes the unique properties of central nervous system (CNS) blood vessels which allow them to stringently regulate the extracellular environment of the surrounding neural tissue. While it is known that different brain regions are made up of diverse cell types and execute distinct functions, it is unknown if the BBB exhibits varied features in functionally distinct regions to regulate specific neural circuits. Since little is known about heterogeneity of the BBB, this project will address this gap in knowledge and pursue the question of whether regional specializations of the blood-brain barrier may regulate local circuit function and behavior. This will identify if the BBB could be targeted to modulate specific behaviors, including reward and addiction. Single cell RNA sequencing revealed that there is significant inter-regional heterogeneity of the BBB via differential gene expression in endothelial cells (ECs). While the canonical BBB genes that contribute to restrictive barrier properties were expressed in ECs across all brain regions, many genes involved in transport, signaling, and metabolism were found to be enriched in ECs of specific brain regions. For instance, Stra6, a transport regulator of retinoids, is remarkably enriched in the nucleus accumbens shell (sNAc) and the ventral cochlear nucleus (VCN) compared to other areas of the brain. The sNAc is particularly of interest because its function in reward and reinforcement learning implicates the region in drug addiction. Due to the prevalence and severity of addiction, it is urgently important to find new targets for treating this major public health issue by asking if the BBB, via Stra6, plays a role in regulating addiction. The proposed study will address this critical need by determining (1) if the regional specificity of Stra6 at the BBB is necessary for the development or adult function of the sNAc, (2) by what molecular mechanisms Stra6 regulates sNAc function, and (3) if Stra6 is required for learning addiction behavior. This proposal will test the hypothesis that Stra6 concentrates retinoids within the sNAc, thereby dynamically regulating dopamine receptor levels and thus the adult function of the sNAc in both spatial learning and addiction. The importance of Stra6 at the BBB will be tested using conditional, EC-specific Stra6 knockout mice to excise Stra6 during development or in adulthood. These aims will also test whether elevating dietary Vitamin A, the source of retinoids, can increase retinoid levels in the absence of Stra6 and rescue sNAc function. The central goals of this proposal are to gain deeper understanding of the BBB’s contribution to brain function, specify mechanisms of addiction, and assess what factors might increase individual susceptibility to drug addiction.

NIH Research Projects · FY 2026 · 2021-09

Epilepsy is the fourth most common neurological disease, costing the healthcare system approximately $15.5 billion annually and negatively impacting quality of life. The incidence and prevalence of epilepsy peaks over the age of 55—a group that is particularly vulnerable to accelerated cognitive and brain aging, placing them at increased risk for progressive neurodegenerative disorders, including Alzheimer's disease (AD). Given that the most rapidly growing segment of the U.S. population is adults over the age of 55, the number of older adults living with epilepsy will dramatically increase over the next several decades, presenting a major public health concern. Therefore, there is a critical need to characterize cognitive and brain aging in older adults with epilepsy, identify underlying mechanisms of accelerated aging, and target modifiable risk factors that would prevent or mitigate cognitive decline and progression to dementia. We propose the first longitudinal, multi-site investigation of cognitive and brain aging in older adults (55-90 years) with epilepsy in efforts to identify vascular, genetic, biomarker and demographic risk factors for accelerated aging. We will accomplish this goal by obtaining state-of-the-art neuroimaging, comprehensive neuropsychological, vascular risk, and genetic/biomarker data on 100 patients with temporal lobe epilepsy (TLE) and frontal lobe epilepsy (FLE) from three geographically and racially/ethnically diverse epilepsy centers. We will follow these patients longitudinally, examine their imaging and cognitive trajectories over 5 years, and compare their trajectories to 100 patients with mild cognitive impairment (MCI) and 100 normal aging controls. We will then examine the influence of vascular, genetic (apolipoprotein 4), and cerebrospinal fluid biomarker (i.e, amyloidβ and tau) risk profiles on cognitive decline and identify baseline factors that increase risk for progression to dementia. Our scientific premise is that older adults with focal epilepsy will show age-accelerated cognitive and brain aging comparable to that seen in MCI. We propose that elevated vascular risk and the presence of AD- associated pathology will underlie the association between accelerated brain aging (i.e, regional atrophy, white matter injury, and hypoperfusion) and cognitive decline in vulnerable patients. These goals are aligned with the 2014 NINDS Benchmarks for Epilepsy Research, which prioritize limiting or preventing adverse consequences of seizures and their treatment across the lifespan. They are also aligned with the AD/Alzheimer's Dementia Related Dementias (ADRD) research goals of identifying risk factors (i.e., seizures) for progression to dementia. The current project has strong implications for public health because it aims to identify individual predictors of cognitive decline that could help to prevent disabilty and progression to dementia, which would have an immediate and sustained impact on patient care. Furthermore, this grant will explore the bi-directional link between AD and epilepsy, would could lead to therapeutic opportunities for both diseases and other disorders of aging.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT The human brain is adept at constructing coherent perceptual experiences, despite an environment overflowing with more information than can be processed at any given moment. Out of necessity, top-down cognitive processes, such as attention and working memory, play a key role in regulating what we can and cannot perceive. What neural computations drive this regulation of perception? While computational models have been proposed to account for the effects of attention and working memory on visual processing, there is a lack of empirical evidence testing these models in humans. This project will bridge this gap by pairing computational modeling with non-invasive neuroimaging techniques, such as functional magnetic resonance imaging (fMRI) and electroencephalography (EEG), to study the brain mechanisms underlying attention, perception, and working memory. By using these techniques to track the neural fate of visual information across human visual cortex during selective attention tasks, we will reveal the specific mechanisms by which top-down processes improve our ability to see. In the long-term, development of a computational model will better quantify the top- down regulation of visual processing – information key to vision scientists and basic researchers investigating impaired cognition in patient populations. The aims of this project align with the mission of the NINDS and the BRAIN Initiative, building innovative multi-modal neuroimaging approaches to understand the brain mechanisms that underlie attention, perception, and working memory, with potential for guiding the diagnoses and treatment of related disorders.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Lung is one of the largest internal sensory organs. Chronic exposure of the lung to allergens or other irritants has been shown to influence stress, anxiety, depression, and dementia. On the other hand, practices that modulate respiration including deep breathing in meditation or Qi Gong are linked to improved central nervous system (CNS) health. However, the functional neuroanatomical connections between brain and lung at the molecular and cellular level remain largely unknown. This R01 application proposes to test the hypothesis, using mouse models, that allergen-induced signals detected by interoceptors in the lung are transmitted via vagal afferent neurons to nucleus tractus solitarius (NTS) (aim 1), and then from NTS to other central integrators in the brain, potentially including paraventricular nucleus (PVH) in the hypothalamus (aim 2), and lastly from dorsal motor nucleus of the vagus (DMV) to descending spinal cord efferents that project back to the lung (aim 3) to sense and regulate airway hyperresponsiveness. The aims will incorporate cutting-edge cell-type specific circuit and viral tracing techniques, lightsheet 3-D imaging, and candidate multiplex RNA in situ and unbiased snRNAseq approaches to map the neural circuits and identify the signature of signal-activated neurons in peripheral ganglia, NTS and other brain regions. In addition, they will combine chemogenetic, optogenetic and toxin-based approaches with neural activity readouts including cFOS, in vivo calcium imaging, electrophysiology, and lung physiology to assess gain and loss of functional effects of key lung, vagal, spinal and brain connections.

NIH Research Projects · FY 2024 · 2021-09

Atrial fibrillation (AF) is the most common arrhythmia, affecting approximately 35 million people worldwide. During AF, the heart's two upper chambers (the atria) beat weakly and irregularly creating regions of slow flow (blood stasis) where clots may form. Clots preferentially form within the left atrial appendage (LAA) and can travel to the brain resulting in stroke. The risk of embolic strokes in AF patients is reduced with anticoagulation medications but, due to the associated increased bleeding risk, these medications are not recommended for all AF patients. Determining if anticoagulation is beneficial requires assessing if patients' risk of stroke outweighs the bleeding risk. However, current methods to risk-stratify AF patients for stroke are not personalized and, for a large number of patients, leave uncertainty as to whether anticoagulation is beneficial. The main objective of this project is to develop novel CT imaging analyses to quantify the personalized risk of LAA thrombosis in AF patients. Our scientific premise is that blood stasis is a key ingredient of thrombosis because it permits thrombogenic reactive species to interact and initiate clot formation. Our preliminary data suggest the spatio- temporal dynamics of blood flow and wall motion in the atrium and LAA strongly correlate with thrombus formation. Our approach consists of three specific aims. In Aim 1 we will develop and validate a computational framework to quantify left atrial blood stasis by 4D CT imaging of atrial kinetics combined with computational fluid dynamics (CFD). We will develop image processing algorithms to quantify left atrial kinetics based on time- resolved CT scans, including the spatio-temporal dynamics of contrast opacification, imaged wall motion, and the non-Newtonian rheology of blood flow in the LAA. In Aim 2 we will establish the relationship between 4D atrial kinetics by multi-heartbeat contrast CT and blood stasis using CFD, in order to facilitate the clinical translation of stasis mapping by CT alone. We will also perform the first rigorous analysis of how uncertainty caused by imaging resolution, modeling assumptions, and physiological variability propagates into predictions of LAA blood stasis. In Aim 3 we will perform an outcome-based clinical pilot study to develop a personalized image-based thrombosis risk score. We will acquire CT data in patients with a history of LAA thrombus or AF- associated stroke and a matched comparison group of AF patients with no history of thrombosis. We will use this unique data set to develop a patient-specific image-based risk score incorporating CT contrast opacification analyses with functional and geometric parameters. Our team includes a cardiologist with a physics background specializing in imaging, an engineer with expertise in CFD analysis, and an engineer with expertise in quantitative analyses of cardiac imaging. Our translational goal is to provide clinicians with a novel image-based tool for personalized risk stratification of patients with atrial fibrillation to guide anticoagulation decisions and improve outcomes.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Somatic copy number amplification (SCNA) of tumor promoting oncogenes, and focal copy number amplifi- cations specifically, are a major driver of cancer pathogenicity. Recent results have revealed that that focal oncogene amplification is mediated to a large extent by extrachromosomal DNA (ecDNA) i.e., large (1.3 Mb on average), highly amplified, oncogene-containing circular molecules that occur in nearly 25% of cancers across all sub-types, but rarely in normal cells. Unresolved questions regarding the formation, evolution, het- erogeneity, and pathogenicity of ecDNA are becoming central to uncovering vulnerabilities that can be targeted for diagnostics and therapy. The proposed project will enhance and disseminate “Software and algorithms for elucidating the structure, function, and evolution of extrachromosomal DNA.” Specifically, we will (1) de- velop CAPER (a Community Accessible Pipeline for EcDNA Reconstruction) by leveraging the GenePattern ecosystem to provide an easy point and click interface to running the CPU, memory and storage heavy soft- ware; (2) design and implement novel algorithmic improvements to the CAPER work flow, including support for long-reads and integration of Omics data; and, (3) enable the broad adoption of CAPER through strategic collaborations, outreach and education.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Abnormalities in right ventricle (RV) development are associated with congenital heart disease (CHD). Although developmental origins and initial specification of the RV has been a subject of intense study, critical molecular pathways involved in subsequent RV growth and expansion at later stages following chamber formation remain to be explored. Our analysis of mice with constitutive cardiomyocyte (CM)-specific knockout (cKO) of Nedd4 revealed a unique RV-specific phenotype. Deletion of Nedd4 during early embryonic stages by Xmlc2-Cre, which specifically expresses in early developing CMs from E7.5, resulted in dramatic RV dilation at postnatal stages, with defects observed at E14.5. Conversely, the LV, as well as the pulmonary vessels, appeared normal. NEDD4 is a HECT E3 ubiquitin ligase highly expressed in CMs. The severe RV-specific abnormalities in Nedd4 cKO highlight the importance of NEDD4 in RV development. To explore molecular mechanisms by which loss of NEDD4 in CMs resulted in RV development defects, we performed proteomics analysis of cKO and control ventricles at E14.5. Differentially expressed proteins were enriched in pathways involved in cellular membrane organization, vacuolar transport and vesicle-mediated transport. To determine whether loss of Nedd4 disrupts cellular membrane organization, we performed TEM analysis on cKO and control hearts at E14.5. Intriguingly, a massive expansion of endoplasmic reticulum and perinuclear space accompanied by large cellular vacuoles was observed in RV tissues of cKOs, while the LV displayed normal ultrastructure, suggesting that Nedd4 is essential for cellular membrane organization in RV CMs. To determine the ongoing CM-specific requirement for NEDD4 in RV development at different stages, we generated tamoxifen inducible CM-specific Nedd4 knockout mouse models to ablate Nedd4 at embryonic, perinatal or adult stages. Our preliminary data revealed that ablation of Nedd4 in either postnatal or adult CMs did not result in the dilated RV phenotype observed in Nedd4 cKOs, suggesting a specific requirement for Nedd4 in embryonic CMs, although the critical time window during which Nedd4 controls RV development, and detailed molecular mechanisms by which it does so remain unknown. The unique RV phenotype of Nedd4 cKO mice will allow us to uncover RV-specific pathways required for heart development, and to model the impact of RV dysfunction on cardiac performance, of relevance to CHDs associated with RV dysfunction. We will test the hypothesis that Nedd4 plays essential roles in CM subcellular membrane organization and RV development. Our Specific Aims are: 1) To elucidate the role of Nedd4 in RV development, as well as the impact of congenital RV defects on cardiac performance, by histological, physiological, biochemical, and molecular analyses of constitutive Nedd4 cKO mice; and 2) To determine the critical window during which Nedd4 is required for RV development by analysis of Tnnt2MerCreMer:Nedd4 inducible CM-specific knockout (ticKO) mice.

NIH Research Projects · FY 2025 · 2021-09

Fall-related injury in the elderly carries a 20% mortality rate, and is the sixth leading cause of death in this population. Age-related dysfunction of gravity receptors within the vestibular system is highly correlated with these elderly falls, and significant age-related degeneration is associated with nearly all types of vestibular cells. The overarching goal of this study is to analyze human and mice genome-wide association studies (GWAS), vestibular-specific human and mice genomic expression, and single-cell sequencing of specific sites in the vestibular system, then test identified genes and related pathways in the lab. Our objective is to characterize the genomics related to age-related imbalance for future prevention and treatment. The central hypothesis is that by this analysis, we can identify anatomic and physiologic sites relevant to the balance system that is common to both species. Our rationale is that by this analysis, we can better focus on relevant genes and pathways for lab testing and ultimate therapeutic intervention. Building upon a small GWAS on elderly falls that correlated human DCC and PTK2 genes in the same pathway as Dcc identified in the Hybrid Mouse Diversity Panel (HMDP) GWAS, our specific aims will be: 1) a. Perform GWAS in humans based on a dizziness/falls phenotype in a meta-analysis of large datasets; b. GWAS in mice based on a behavioral and gravity sensor function phenotype in the HMDP; 2) a. Perform RNA-Sequencing on vestibular tissues from mice and human surgical specimens; b. Single-cell RNA-Seq on individual tissues; c. Compare identified genes and pathways via computational methods to assess translation of pathways from the mouse to human balance system; and 3) Perform functional testing for the top candidates defined in Aim 2 using knock-out/knock-in mice. Multiple innovations of this project include: 1) the first GWAS of gravity receptor function in aged mice and in elderly humans, 2) a comprehensive catalogue of genes and pathways involved in vestibular functional variation with inter-species comparison, as part of FAIR (findable, accessible, interoperable, reusable) Compliance, 3) in vivo validation in mouse models and an analysis of these candidates in available human cohorts, and 4) future potential for targeted therapies. Our outcome is the first comparative GWAS of the balance system between animal models and humans. The impact of this work will be to lay a firm foundation for development of targeted treatment of the balance system to diminish falls in the elderly.

NIH Research Projects · FY 2024 · 2021-08

Abstract Although cardiovascular disease (CVD) has traditionally been regarded as a higher risk condition in men, it is also the leading cause of death in women. The impact of sex and gender on the pathophysiology of CVD has emerged as an important clinical issue but its molecular mechanisms are poorly understood. The intersection of sex, gender and CVD is underscored by recent observations that transgender individuals undergoing gender affirming sex hormone therapy (GHT) are at increased CVD risk. Sex-differentiation of insulin resistance, a major CVD risk factor, may underlie these observations since insulin resistance arises from adipose, a sexually dimorphic tissue that is highly susceptible to sex hormones. Unfortunately, options to treat insulin resistance- mediated CVD risk in a sex- and gender-relevant fashion are limited particularly in women. For example, thiazolidinediones (TZDs), the only pharmacologic class that specifically targets insulin resistance in adipose, increase the risk of osteoporotic fractures in post-menopausal women. A molecular understanding of the roles of chromosomal sex, sex hormones and gender on the development of insulin resistance is needed to enhance screening and develop new therapeutic options for CVD in women and transgender individuals. One approach to identifying the molecular determinants of insulin resistance specifically operational in women is to conduct genetic association studies (GWAS) of insulin resistance stratified by sex. However, isolating the relative impacts of chromosomal sex, sex hormones, and gender and establishing a mechanistic relationship between phenotype and identified genetic variants remains a major challenge. Here, we propose to: 1) Utilize an integrative genomic approach by leveraging the natural “crossover” experiment between sex chromosomes, hormones and gender that occurs in transwomen and men undergoing GHT; and 2) Directly test the impact of perturbing putatively causal genes on CVD risk in women by using high throughput assays for insulin resistance to functionally characterize protein-coding genetic variants identified in 273,000 women. This work will systematically identify sex- and gender-specific insulin resistance genes and for several top- ranked genes, assess the clinical effect on CVD in women by relating genetic variants to function to phenotype.

NIH Research Projects · FY 2024 · 2021-08

PROJECT SUMMARY/ABSTRACT The airway is composed of luminal cells such as club and ciliated cells that moisturize and clean the airway, respectively. They are lined by basal cells that serve as progenitors for luminal cells in normal turnover and injury repair. Proper balance of progenitor and luminal/differentiated cell ratio is critical for airway function. While many genes have been identified that control individual cell fate, knowledge gaps remain in how the ratio of progenitors and differentiated cells are globally regulated. In this study, we will investigate how airway cell ratio is established in development, maintained in homeostasis and restored following injury. Our entry point is Lon protease 1 (LONP1), an ATP-dependent serine protease that functions in the mitochondria matrix to degrade oxidized and misfolded proteins, thereby control protein quality and mitochondria health. Mutations in LONP1 has been identified in congenital diaphragmatic hernia (CDH) patients. CDH carries a high mortality rate associated with lung hypoplasia and pulmonary hypertension. To address if Lonp1 plays a role in lung, we inactivated it in the developing lung epithelium and mesenchyme. While the mesenchymal mutants survived to adult with no discernable phenotype, the epithelial mutants exhibited lung hypoplasia and died at birth. Unexpectedly, these mutants also exhibited a striking increase of basal cells at the expense of club and ciliated cells. Further preliminary data revealed an increase in integrated stress response (ISR) pathway genes, and an increase of KDM6B, a key histone demethylase. In this study, we will investigate the role of this mitochondria factor LONP1 in controlling airway cell fate balance via ISR pathway (Aim 1), chromatin regulators (Aim 2) and in adult airway homeostasis and following influenza- induced injury (Aim 3). Our findings will delineate a novel pathway from a mitochondria protease to ER ISR to nuclear chromatin regulators in the fundamental control of airway cell fate.

NIH Research Projects · FY 2025 · 2021-08

SUMMARY Mexican American women are disproportionately affected by obesity and obesity-related conditions, such as type 2 diabetes. Obesity and diabetes are highly concordant in Mexican American families. Given the younger age of onset of diabetes in women with familial history, targeting mothers and their adult daughters for obesity treatment is warranted. From a family systems perspective, family-level approaches to obesity treatment can improve the adoption and maintenance of weight management behaviors. Including family members in treatment may also serve as a culturally salient intervention strategy as Mexican Americans endorse high level of familism. In contrast to traditional individual-level approaches to obesity treatment, a family-level approach grounded in familism would promote shared goals, collaborative problem solving, and communal coping when treating family members alongside each other. An important construct to consider when working with intergenerational Mexican American families is differences in acculturation, which may translate into differences in attitudes and behaviors. A wider gap in acculturation between parent and child has previously been associated with lower family functioning (e.g., poor communication, high conflict, low cohesion). However, interventions that promote bicultural competence by changing interactional patterns have been effective at improving family functioning. Hence, this study will conduct a randomized control trial testing the efficacy of a behavioral weight management intervention with brief and structured counseling on family functioning. Mexican American mothers and adult daughters (n=118 dyads) will be randomly assigned to receive standard behavioral treatment (SBT) or standard behavioral treatment plus relationship skills training (SBTR). Dyads participating in SBT or SBTR will attend 24 weekly sessions focused on nutrition and physical activity education along with behavior modification techniques. Dyads participating in SBTR will also receive experiential-based relationship skills training that draws from both general family systems concepts and behavioral family/couples therapy approaches to support familism, biculturalism, and communication competencies. The 12-month trial will consist of an intervention phase (1-6 months) and a maintenance phase (7-12 months). Assessments will be conducted at baseline and at the end of the intervention and maintenance phases. The primary outcome is weight loss. Secondary outcomes include treatment adherence (session attendance and self-monitoring records), physiological indicators of diabetes risk (hemoglobin A1c, waist circumference, and body fat percentage), health behaviors (eating and physical activity), psychological well- being (depression and perceived stress), and family functioning (subjective self-report and objective behavioral coding). Dyads in the SBTR group are expected to achieve greater improvements in primary and secondary outcomes than the STB group. Additionally, mediation by family functioning of intervention effects on primary and secondary outcomes will be examined.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY/ABSTRACT Sex differences in the risk of Alzheimer’s disease (AD) and AD pathology burden have been extensively studied; however, little is known about how AD pathology burden relates to clinical symptoms in women versus men. Evidence of a cognitive advantage in the preclinical stage of AD, yet a two-times steeper cognitive decline thereafter indicate that the question of sex differences in the clinical manifestation of AD pathology is an important one. These sex differences have clinical implications in that our established thresholds for AD clinical and biological markers used to diagnose and track disease were typically generated without consideration for sex disparities. If women are better able to maintain what our current cognitive thresholds consider “normal” cognition until a more advanced pathology state than men, then diagnosis of MCI could be delayed, thus limiting the opportunity for early intervention. We hypothesize that sex differences in the clinical translation of AD pathology results from a sex-specific balance of brain-related resilience/risk factors that change with disease stage. Our proposal is particularly innovative in that we will first characterize sex differences in how AD pathology relates to clinical symptoms by disease stage and then examine its neurobiological underpinnings and clinical implications. We will leverage both in-vivo, longitudinal biomarker data from the Alzheimer’s Disease Neuroimaging Initiative (ADNI) and prospective neuropathological data in brain tissue from multiple Alzheimer’s Disease Research Centers (ADRCs). Given their strong ties to AD pathology and the sex differences that our earlier data show, we will examine the brain resilience/risk mechanisms of (1) PET-measured brain glucose metabolism, (2) NMDAR density, a marker of glutamate neurotransmission, and (3) translocator protein 18kDA (TSPO) levels, a marker of microglial activation. Specifically, Aim 1 will utilize ADNI data to examine sex differences in trajectories of cognitive function and their relationship to longitudinal variation in AD pathology (Aβ and Tau) and brain metabolism by AD stage. In Aim 2, we will conduct in vitro autoradiography in hippocampal and cortical brain tissue of 60 normal control, 60 mild cognitive impairment and 60 AD dementia autopsy cases to determine sex differences in plaque, tangle, NMDAR and TSPO density and how they relate to each other and to antemortem cognitive function in each of the three diagnostic groups. In Aim 3, we will take action on these sex differences by generating sex-specific cut-scores for cognitive tests commonly used in MCI/AD diagnostic criteria with the optimal balance of sensitivity/specificity in detecting the presence of clinically-significant levels of AD biomarkers/pathology. The public health benefits of our project would be significant in that by understanding and accounting for sex disparities in our clinical and biomarker approaches to AD diagnosis, we will improve clinical and biomarker approaches to disease diagnosis and tracking in both sexes and possibly identify sex-specific therapeutic targets.

NIH Research Projects · FY 2024 · 2021-08

PROJECT SUMMARY Mutations in genes encoding nuclear envelope (NE) components cause an array of diseases referred to as nuclear envelopathies that often manifest as cardiomyopathies. The NE separates the nucleoplasm from cytoplasm and is composed of outer and inner nuclear membranes. The nuclear lamina (NL) is an extensive network of lamin polymers and associated proteins that are embedded in the inner nuclear membrane (INM). Functionally, NL and INM proteins not only provide mechanical stability to the nucleus but also serve as the anchoring point for chromatin at the nuclear periphery, playing a critical role in chromatin organization and regulation of gene expression via interaction with and modulation of epigenetic machinery components. The LAP2-Emerin-MAN1-domain (LEM-D) family of proteins play an important role in the association between the NL and chromatin. LEMD2 (LEM domain-containing protein 2), is a transmembrane protein located in the INM, involved in nuclear integrity and perinuclear tethering and transcriptional silencing of heterochromatin. Recently, it has been reported that a single amino acid substitution of leucine 13 to arginine (L13R) in LEMD2 leads to autosomal recessive human cardiomyopathy. However, despite its clinical relevance, little is known as to the specific role of LEMD2 in cardiomyocytes (CMs), and mechanisms by which the L13R mutation leads to cardiomyopathy. To address this gap in knowledge, we have generated three mouse models: constitutive CM- specific Lemd2 knockout (cKO), inducible CM-specific Lemd2 knockout (icKO), and LEMD2 L13R knock-in mice. We will also utilize a human induced pluripotent stem cell (iPSC)-derived CM model of the LEMD2 L13R mutation to address the impact of L13R mutation in LEMD2 on human CMs. Preliminary studies revealed that loss of LEMD2 in embryonic or adult CMs is detrimental, and LEMD2 L13R mutation affects cardiac function in a murine model, indicating that LEMD2 plays a critical role in maintaining normal CM structure and function in developing and adult hearts. Thus, we hypothesize that LEMD2-mediated maintenance of NE integrity and/or regulation of heterochromatin tethering and silencing is essential for CM structure and function, and LEMD2 L13R mutation impairs specific aspects of LEMD2 function leading to cardiomyopathy. Accordingly, Our Specific Aims are: 1. To determine the role of LEMD2 in the developing and adult myocardium by analyzing constitutive (cKO) and inducible (icKO) Lemd2 CM-specific knockout mice for heart morphogenesis, structure and function, and the progression of cardiomyopathy; and 2. To elucidate molecular mechanisms underlying cardiomyopathy consequent to the L13R mutation in LEMD2 by detailed analyses of LEMD2 L13R knock-in mice and human induced pluripotent stem cell (iPSC)-derived LEMD2 L13R mutant CMs.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Gross chromosomal rearrangements (GCRs), a type of genome instability, are often seen in inherited and sporadic cancers and are an important driver of malignant progression. For example, hereditary breast and ovarian cancers resulting from BRCA1 and BRCA2 germline mutations suffer from homologous recombination repair (HRR) defects that increase GCRs in model systems. Our previous studies have established a comprehensive network of genome instability suppressor (GIS) genes in yeast and demonstrated in silico that human homologs of these yeast GIS genes are genetically and/or epigenetically altered across many cancer types. From yeast genetic studies, we have also identified and rank-ordered synthetic lethal (SL) partners of non-essential GIS genes. The feasibility of targeting SL genetic interactions for rational cancer therapy has been supported by the application of PARP inhibitors for maintenance therapy of breast and ovarian cancers with BRCA1 or BRCA2 defects. To exploit additional SL interactions for cancer therapy, this proposal aims to leverage the wealth of knowledge on GIS genes and their SL partners developed in yeast to guide the development of therapeutics that target human GIS gene defects that cause GCRs in cancer. Specifically, the proposed studies will focus on the human Flap Endonuclease 1 (FEN1), the homolog of yeast RAD27, which has the highest number of SL-interactions with GIS genes of a variety of functions. FEN1 processes Okazaki fragments during lagging strand DNA synthesis and acts in long-patch base excision repair but is itself non- essential. To develop this nuclease as a target for treating cancers with defects in BRCA1, BRCA2 and other FEN1-SL partner GIS genes, we propose to carry out the following lines of research: AIM 1) To expand ongoing CRISPR-dropout screens and validation studies in cell lines and mice to (a) identify human genes in which defects cause sensitivity to our proprietary FEN1-inhibitors of highly potency and specificity and (b) define FEN1 SL-partner genes as well as cancer omics signatures that can be targeted with FEN1 inhibitors to induce SL; AIM 2) To combine informatics and cell-based validation approaches for a deep-dive into cancer omics and gene essentiality data to identify the spectrum and signatures of cancers amenable to therapeutic targeting with FEN1 inhibitors; AIM 3) To apply an array of genetics-based approaches to investigate mechanisms for acquired resistance to FEN1 inhibitors and to compare resistance to FEN1 versus PARP inhibitors; and, AIM 4) To determine the effects of FEN1 inhibition on DNA replication, fork stability, and chromosome integrity in BRCA1 and BRCA2 mutant cells to test the hypothesis that FEN1 inhibitors induce irreparable damage to replication forks in HRR-deficient cancer cells to cause lethality. These mechanistic studies will be extended to other validated FEN1-SL partner genes. These projects will greatly accelerate the development of FEN1 inhibitors to target cancer genome instability and will establish an experimental platform for evaluation and development of other potential therapeutic targets to be identified by the proposed research.

NIH Research Projects · FY 2025 · 2021-08

Abstract Brain somatic mosaicism (BSM) refers to the accumulation of mutations within any of the billions of cells in the human brain, which can occur from embryogenesis through adulthood. The extent, impact and mechanisms of BSM on brain disease remain poorly understood. Prior work from the Brain Somatic Mosaicism Network (BSMN), on which the PI served, made critical breakthroughs in reliability of mosaicism detection, but also raised new questions, including the degree to which BSM exists in the healthy brain, and the mechanisms by which BSM mutations explain disease. Focal cortical dysplasia (FCD) is associated with substantial neuropsychiatric disability, and is the most common cause of intractable epilepsy in childhood. Neuropsychiatric features are seen in 15-59% of patients 5-7, and neuropathologically shows disrupted neurogenesis, migration, differentiation, and altered neural excitability. We and others previously identified mosaic mutations in the mTOR pathway in a minority of FCD cases, but most cases remain unsolved, and fundamental mechanisms are lacking. We hypothesize that: 1] FCD mutations are similar to neutral somatic mutations in their patterns and distributions, dictated by developmental processes, but differ in their functional effect. 2] BSM patterns, allelic fractions (AFs) and allele sharing between cells can reconstruct cellular lineages and migratory histories. 3] Study of FCD resected tissue can uncover novel causes of disease that would not be tolerated if present in every cell. 4] BSM modeling in mouse can unravel disrupted signaling networks of complex mosaic mutations. Our preliminary data shows: 1] From a post-mortem control cadaver, we validated 259 somatic variants using 300X genome sequencing, and started to use these variants as ‘barcodes’ to reconstruct lineage histories. 2] Deep sequencing from 314 FCD patient brain resections identified 12 new candidate genes, highlighting signaling and synaptic dysfunction, and a novel ‘two-hit’ disease mechanisms. 3] We established in utero mouse electroporation models to assess putative FCD variants as gain or loss of function, and to assess effects of ‘single-hit’ and ‘two-hit’ mutations. We propose three aims: 1] From control cadavers, we will reconstruct cell lineage across anatomical domains using BSM as barcodes. 2] With this lineage information, we will study the origins of BSM mutations in FCD, by recruiting new patients, performing both targeted and unbiased sequencing, and identifying novel causes. 3] We will functionally validate putative deleterious alleles in animal models for both ‘single-hit’ and ‘two-hit’ causes. The goal is to achieve a mechanistic understanding of the extent of BSM in control individuals, to reconstruct neural lineages and to identify novel mechanisms in developmental brain disease.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY / ABSTRACT One in five women and one in fourteen men in the US have been raped, with four out of five rape survivors reporting that alcohol or substance was involved at the time of the violence. Current prevention efforts are hampered by a limited understanding of the gendered and developmental origins of these health crises. Innovative epidemiologic science that integrates psychological and developmental determinants of sexual violence and alcohol use with cutting edge social network and social norms research is critical for advances in prevention efforts. This Mentored Research Scientist Development K01 Award is designed to increase the candidate’s capacities to undertake innovative social network analyses to provide important insights into the gender socialization processes that occur in adolescence and contribute to alcohol use and sexual violence behavior across the lifecourse. The proposed activities will take place alongside a strong and dedicated training committee comprised of globally renowned experts and will extend the candidate’s existing expertise in social epidemiology to include: social and developmental psychology, adolescent alcohol epidemiology, longitudinal social network analysis, and multilevel structural equation modeling. Research activities will involve social network and structural equation modeling techniques using secondary data from a nationally-representative prospective cohort study. Three specific aims are proposed: Aim 1) Longitudinally examine the relationship between individual gender expression, alcohol use, and sexual violence; Aim 2) Assess contribution of normative gender expression within peer groups to variation in alcohol use and sexual violence outcomes; Aim 3) Identify latent classes of adolescents at greatest risk for alcohol use and sexual violence, and the social network processes (selection and influence) that moderate risk. Research significance includes: a) identification of novel and developmentally-specific mechanisms that explain sex-disparities in alcohol use and sexual violence in adolescence; b) identification of typologies of adolescent peer groups at highest risk for both alcohol use and sexual violence. These findings will provide new avenues for prevention efforts to address the developmental origins of sex-disparities in alcohol use and sexual violence. Research innovations include: a) social network algorithms to identify peer groups, used within multilevel models; b) a novel, validated measure of gender expression; c) an integrated multilevel lifecourse framework of adolescent socialization processes linked to alcohol use and sexual violence. Findings from this work will inform a future survey-based R01 among younger adolescents to further elucidate the social psychological processes and normative environments in adolescent peer groups that impact alcohol use and sexual violence, as modified by racial, sexual, and gender minority status. This work responds to the NIAAA Division of Epidemiology and Prevention Research’s (DEPR) Strategic Plan, which underscores the need for research focused on prevention and youth.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT: Hepatocellular carcinoma (HCC) is caused by hepatitis virus HBV/HCV, non-alcoholic steatohepatitis (NASH), and alcoholic liver disease (ALD), which typically progress from liver fibrosis, to cirrhosis and cancer. Our preliminary data demonstrate that genetic deletion of IL-17 signaling in steatotic hepatocytes significantly attenuates the development of HCC in ALD-injured mice, suggesting that IL-17 signaling is a target for anti- HCC therapy. Our central hypothesis is that IL-17 signaling regulates chemokine production, de novo lipogenesis, and TNFRI expression/turnover in steatotic hepatocytes. IL-17 signaling promotes ALD- and NASH-induced HCC via activation of TNF/TNFRI-SREBP1/2-DHCR7-cholesterol synthesis, and suppression of ARTS-1/NUC2-dependent TNFRI exocytosis. The goal of the study is to characterize the mechanism by which IL-17A/IL-17RA signaling regulates responses in metabolically injured hepatocytes, and to compare the pathways of IL-17 signaling in the experimental models of ALD- and NASH. Strategy: Responses to IL-17 signaling will be compared side-by side in ALD- and NASH-injured WT and hepatocyte-specific IL-17RA knockout mice with HCC. We determine if IL-17 signaling is similarly activated in NASH- and ALD-injured hepatocytes. We determine if blocking of IL-17 signaling in steatotic hepatocytes is sufficient to suppress HCC in the metabolically injured liver. Specifically, the role of IL-17 in the pathogenesis of DEN- or (Mup-uPA)- induced HCC in ALD- and NASH-injury will be studied in WT and hepatocyte-specific IL-17RA knockout mice (IL-17RAΔHep mice). Development of HCC, inflammation, steatosis and liver fibrosis will be across all groups of mice. Mutagenesis of WT and IL-17RA-deficient AFP+YAP+ HCC, and responses of steatotic hepatocytes to IL-17A will be characterized. Specifically, we determine if chemokine secretion, cholesterol synthesis are suppressed in metabolically injured IL-17RA-deficient hepatocytes (AIM1). We will test a novel hypothesis by which IL-17 signaling facilitates TNF/TNFRI-Caspase2-SP1-SREBP1/2-DHCR7-dependent cholesterol synthesis in steatotic hepatocytes via blocking ARTS-1-NUCB2-regulated TNFRI exocytosis (and possibly IL- 6, IL-1RII) thereby prolonging TNF (IL-6, IL-1) signaling and promoting alcohol-induced HCC (AIM2). Our findings will be translated into humans by characterization of IL-17RA-TNFRI-signaling pathways in archived human livers from HCC patients with ALD. We will test if therapeutic blocking of the key IL-17 signaling molecules (IL-17RA, TNFRI, ARTS-1, and DHCR7) specifically in hepatocytes using N-acetylgalactosamine (GalNAc)-conjugated antisense RNA oligonucleotides (ASOs) can effectively suppress steatosis, fibrosis, and HCC in WT mice with NASH and ALD (AIM3). If proven, hepatocyte-specific blocking of IL-17 signaling using GalNAc-ASOs can provide a new strategy for HCC treatment in ALD and NASH patients.

NIH Research Projects · FY 2024 · 2021-08

Description/Abstract Schizophrenia and related psychotic illnesses are neurodevelopmental disorders with evidence of pathological changes beginning in utero; neuromotor and neurocognitive abnormalities in the premorbid period; subsyndromal psychotic symptoms in the prodromal period of illness (also called clinical high risk, CHR); and full manifestation of a psychotic syndrome during late adolescence or early adulthood. CHR research over the past 2+ decades, has provided (i) important insights into risk factors for later conversion to full psychotic illness, (ii) the development of a “Psychosis Risk Calculator”, (iii) biomarkers linked to psychosis riskand (iv) evidence of dynamic brain changes that are likely present before the onset of illness and continue to evolve into the first episode psychosis (FEP), as well as into more chronic forms of psychosis. Despite these advances in our understanding of the CHR state, the longer-term outcomes (5+ years), and the trajectory of diagnoses, symptoms and psychosocial function have been seldom investigated in this population. Meta- analyses show that 20-30% of identified CHR individuals develop psychosis within 2 years. Little is known about what type of psychosis (affective versus non-affective) "declares itself" after evidence of the initial conversion to psychosis, the rate of later psychotic conversion (i.e. post 2-3-year follow-up periods) or risk factors that might predict a later onset of psychosis. The majority of individuals who meet CHR criteria do not develop overt psychosis within 2 years but demonstrate outcomes ranging from complete remission to continued symptoms and functional impairment, at least within this relatively short time frame. Longer-term follow-up of CHR individuals provides a unique and rare opportunity to investigate the full trajectory of illness from CHR -> First Episode -> Chronic Illness, in addition to longer-term outcomes in symptomatic individuals who did not develop psychosis within 2 years after ascertainment. Substantial evidence already exists for multiple biomarker abnormalities in CHR subjects. Specifically, CHR youth show deficits in neurocognition, regional cortical gray matter, event related potential (ERP) amplitudes as well as higher polygenic risk scores (PRS), inflammatory markers and cortisol relative to comparison subjects. Biomarkers also predict who will convert to psychosis and functional outcomes at 2 years. However, it is not known whether these biomarkers predict longer term conversion and outcomes. The Specific Aims are to 1) Perform long-term (5-20 year) diagnostic, symptom and functional assessments of up to 2000 individuals who previously met CHR criteria, some of whom converted to psychosis, across 9 academic centers. 2) Determine the 5+ year psychotic conversion rate of CHR individuals and use baseline demographic, clinical, functional, neurocognitive and biomarker data to predict longer term functional and diagnostic outcomes of individuals who convert to psychosis and 3) Investigate the long-term diagnostic and functional outcomes of CHR individuals who do not convert to psychosis and test whether outcomes are influenced by treatment and substance use.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Mounting evidence links the gut microbiome – the modifiable “second genome” consisting of trillions of diverse microbes that inhabit the human gut – with type 2 diabetes mellitus (T2DM) in humans. Studies using fecal transplant in mice have raised the central hypothesis that changes of the gut microbiome and the biochemical by-products originated from these microbes may be key modulators of development of T2DM. To date, however, knowledge of the specific microbial species that drive the development of T2DM in humans remain very limited. Human studies of the gut microbiome in T2DM have largely been cross-sectional and confounded by reverse causality. Indeed, many of the gut microbiome changes observed in human T2DM have been found to be a consequence of the disease or treatments, including the medication metformin, rather than a cause of T2DM. In addition, the circulating metabolites derived from the gut microbes that contribute to the development of T2DM remain to be discovered. In this Early Stage Investigator NIH R01 application, we propose the largest prospective study of the role of gut microbiome in T2DM using a Finnish cohort of 6921 individuals with fecal and plasma samples collected in 2002 and over 15 years of subsequent clinical follow up. This proposed study brings together a diverse team with deep expertise in the human microbiome, mass-spectrometry based metabolomics, computational biology, statistical epidemiology, and diabetes pathobiology, to specifically address the key biological and clinical questions regarding the role of gut microbes and microbial derived metabolites in the development of incident T2DM in humans.

NIH Research Projects · FY 2025 · 2021-08

Abstract The main goal of this project is to determine the contribution of human microglia in the establishment of early neural networks during development in healthy and autistic conditions. Although the exact cause of Autism Spectrum Disorders (ASD) remains unclear, epigenetic, genetic and environmental factors are at play. Given that ASD is a complex multifactorial disorder and that epigenetic modifications have been shown to control microglial phenotypes/plasticity, we hypothesized that microglial epigenetic signature might influence neuronal development. Given that microglia originate in the periphery and later invade the brain, they are most likely the first brain cell type to be exposed to an environmental factor or at least be impacted by the environmental factor given their role as gate keepers of the brain. Therefore, a better understanding of the genetic, environmental or a synergistic impact of both, will pave the way to a better understanding of human neurodevelopment and human microglial roles during this process yielding to the discovery of novel therapeutic targets and efficient therapies for a broad range of neurological disorders including ASD. Thus, with this project, we aim to establish whether and how human microglia interfere with neural network establishment and if high-risk ASD epigenetic genes could alter their function and their role during human neurodevelopment. Based on our preliminary data we propose the following specific aims are: Aim 1: Determine the role of healthy human microglia on healthy brain cortical organoids (BCO), Aim 2: Measure the impact of microglia carrying ASD mutations on BCO development and function, and Aim 3: Impact of environmental ASD-risk factors in combination with underlying genetic predisposition: the two-hit hypothesis. Here we will test the isolated and additive effect of ASD-related environmental factors on the function of microglia and its impact on BCO physiology.