University Of Southern California

NIH Research Projects · FY 2026 · 2023-08

Abstract Age-related macular degeneration (AMD) is the leading cause of severe visual impairment in people over age 50 in developed countries [1,2]. Transplantation of stem cell derived retinal pigment epithelium (RPE) is currently a promising method to treat retinal degeneration and advanced non-neovascular AMD (NNAMD) [3- 6]. Many protocols have been developed for the derivation of RPE from pluripotent stem cells from human embryonic stem cells (hESC) or human induced pluripotent stem cell (iPSC) [7-12]. The quality control of donor cells is a basic requirement for cell production in clinical trials. Stem cell residues and chromosome number variation during long-term culture must be tested before clinical use. However, quality control for stem cell residues (pluripotency) and stem cell tumorigenicity is not trivial. Physically removing cells from an RPE monolayer during culture will result in hypotrophy of the monolayer due to epithelial-mesenchymal transition (EMT) and wound healing. There is an unmet need in the molecular profiling of RPE implants with spatial RNA sequencing (RNA-seq). A focused ultrasound (FUS) offers a solution to this unmet need, as it can produce ejection of cells via cell-containing liquid droplets from a solid surface with minimum impact on the edges surrounding the ejection spot. Ultrasound propagates through liquid and solid, and the FUS transducer does not have to be in physical contact with the substrate where cells are grown. The number of cells that are ejected by a FUS transducer depends on the focal size of the FUS, which can be very small, and is very precise and repeatable. Further, it is low-cost and effective for isolating tens to hundreds of single-cells in parallel through an array of transducers. To satisfy the unmet need and allow realization of RPE cell therapy for AMD, we propose to use self-focusing acoustic transducers (SFATs) for damage-free, cell-containing droplet ejection from RPE monolayers grown on thin solid scaffolds for spatial single-cell RNA-seq. Besides for quality control in RPE implant production, the SFAT’s unprecedented capability of on-demand ejection of microparticles or cells (of tens - hundreds of microns in diameter) will open up many new possibilities in spatial molecular cell study, gene transfection, juxtaposition and manipulation.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY The goal of this grant proposal is to advance the preclinical development of a novel macrocyclic peptide, Orynotide™ MTD12813, for treatment of multidrug resistant (MDR) Gram negative bacterial infections, with the initial focus being on infections caused by carbapenem-resistant Enterobacteriaceae (CRE). The emergence of infections by multiple CRE pathogens has created an urgent public health threat, because carbapenems are drugs of last resort for infections caused by an increasing fraction of MDR bacterial pathogens. Just two species, Klebsiella pneumoniae and Escherichia coli, cause an estimated 140,000 nosocomial infections per year in the United States alone, and many are carbapenem resistant. There is global consensus that new preventive and therapeutic strategies are urgently needed to combat the growing problem of MDR bacterial infections. The applicants, leaders in the field of θ-defensin biology, are responding to this need by developing Orynotides, a new class of host-directed antimicrobial macrocyclic peptides bioinspired by the structural and biological properties of theta (θ)-defensins, macrocyclic peptides expressed exclusively in Old World monkeys (but not humans). Exploiting the pleiotropic host defense properties of θ-defensins, we produced a library of novel Orynotides that includes several compounds that are highly effective in MDR Gram negative septicemia models. Hit-to-lead studies identified MTD12813 as the lead Orynotide candidate for preclinical development as a first- in-class immunotherapeutic agent for MDR Gram negative infections. In the mouse peritoneal sepsis model, single dose administration of MTD12813 is highly effective (enhanced survival with concomitant bacterial clearance) against multiple strains of CRE-K. pneumoniae and CRE-E. coli, and additionally was shown to be effective in septicemia caused by MDR Acinetobacter baumannii. Consistent with the range of pathogens against which MTD12813 is active in vivo, we showed that the peptide’s mode of action is immunotherapeutic, promoting host-mediated bacterial clearance, stimulating phagocytosis and neutrophil recruitment, while modulating levels of otherwise dysregulated proinflammatory cytokines. These data indicate that MTD12813 is a novel immunotherapeutic agent effective in the treatment of Gram negative bacterial pathogens. The peptide is readily manufacturable ( ~1.5 g on hand), highly stable in human plasma and whole blood, resistant to bacterial proteases, and well tolerated when administered by numerous routes. In the proposed studies, we will advance the preclinical characterization of MTD12813. Aim 1 studies will include production of GLP MTD12813 and other critical reagents, pharmacokinetic (PK) and PK/pharmacodynamic analyses, and ADME studies. Aim 2 will focus on illuminating mechanism(s) of action at the cellular and molecular level. Aim 3 objectives will include preclinical non-GLP safety and toxicokinetic studies in rats and beagle dogs, evaluation of immunogenicity, development of an antidrug antibody assay, and culminate with formal GLP safety studies. The goal of these aims is to advance the preclinical development of MTD12813 to IND filing with the FDA.

NIH Research Projects · FY 2025 · 2023-07

Project Summary Myeloid immune suppression, driven by macrophages and monocytes, is a known mechanism of intrinsic resistance to immune checkpoint inhibition (ICI), which has provided significant and durable responses in patients with advanced cancers, but not in most breast cancer patients. Our laboratory has been instrumental in demonstrating that targeting myeloid immunosuppression via decreasing the function of immature myeloid derived suppressor cells (MDSCs) is a novel and important strategy to sensitize the tumor microenvironment (TME) and improve the response to ICIs in breast cancer. We published two preclinical studies demonstrating that entinostat, a class I histone deacetylase inhibitor, decreased MDSC suppression of T cells; furthermore, combining entinostat with the ICIs nivolumab and ipilimumab improved survival in murine models of breast cancer. This provided a strong rationale for our Phase I clinical trial (NCI-9844) where we determined the recommended phase 2 dose (RP2D) for this treatment, given to patients with advanced solid tumors, including 10/33 with breast cancer, as a two-week pre-treatment with entinostat, followed by the combination of entinostat + nivolumab + ipilimumab. Furthermore, an overall response rate (ORR) of 30% was observed in an expansion cohort of 20 patients with advanced breast cancer, who received the RP2D. Nevertheless, we have yet to understand how entinostat decreases myeloid suppression to achieve TME sensitization that leads to this striking response to ICIs. Preliminary data in preclinical models demonstrate that entinostat 1) decreases suppressive function of intra- tumoral MDSCs; 2) decreases the activation of the STAT3-NFkB-AP-1 signaling axis; and 3) alters the phenotype and/or infiltration within the TME of other myeloid cells, including tumor associated macrophages (TAMs), and dendritic cells (DCs). Preliminary evaluation of patient samples from our clinical trial confirms these findings. Thus, we hypothesize that entinostat decreases MDSC immunosuppression and shifts the phenotype and function of TAMs, and DCs to collectively sensitize the TME to promote an enhanced response to ICIs. The Specific Aims are: 1) to determine the effects of entinostat on the STAT3-NFB-AP-1 axis decreases MDSC-mediated T cell suppression; and 2) to determine the cellular basis of entinostat-induced sensitization of the TME and how it mediates the anti-tumor response to ICIs. Our unique bidirectional approach combining preclinical studies, e.g., mouse models and mathematical modeling, with hypothesis-driven correlative investigations in patients, and vice versa, strengthens our ability to uncover the molecular and TME mechanisms driving clinical responses. These studies will reveal if changes in myeloid suppression and/or tumor- immune dynamics inform response to therapy or survival, while identification of new biomarkers may improve patient stratification for future trials.

NIH Research Projects · FY 2026 · 2023-07

Abstract Chickens and their embryos provide a valuable research model for the study of developmental biology and human diseases. However, their application has been greatly limited due to the lack of tools to genetically modify chickens and other avian species. Recently, we have developed methods for the derivation of germline competent embryonic stem cells (ESCs) from chickens. The availability of germline competent chicken ESCs opens up a new avenue for producing genetically engineered chickens. In this project, we propose to establish an ESC-based platform for the efficient generation of chicken germline chimeras (Aim 1). We will also optimize methods for the generation of primordial germ cells (PGCs) from chicken ESCs. These chicken ESC-derived PGCs will be used to produce genetically modified chickens (Aim 2). In Aim 3, by taking the advantage of the chicken ESC culture condition we have established, we will develop methods for the generation of induced pluripotent stem cells (iPSCs) from chicken embryonic fibroblasts. Successful completion of these three aims will provide the avian research community with a powerful genetic toolkit for the production of transgenic and gene-targeted avian species and for the conservation of endangered avian species.

NIH Research Projects · FY 2025 · 2023-07

ABSTRACT Sedative-hypnotic "z-drugs" are dependence-forming substances associated with falls, fractures, high-risk parasomnias (sleepwalking, sleep-driving), and cognitive impairment. However, these drugs are popular and are usually prescribed long-term for insomnia, despite guideline recommendations to limit their use and to favor cognitive-behavior therapy for insomnia. Behavioral economic nudges delivered in the electronic health record are a promising way to encourage the use of cognitive-behavioral bibliotherapy and phone apps for insomnia and to reduce high-risk Z-drug prescribing. Nudges are insights from behavioral science that fit into existing clinical workflows, intended to preserve choice but influence decision-making toward desired behavior. We propose the Study in Outpatient Medicine using NUdges to improve Sleep (SOMNUS) in response to NHLBI's Notice of Special Interest (NOT-HL-21-010). The SOMNUS trial will simulate the long-term outcomes of our nudging strategy using microsimulation from our Roybal Center for Behavioral Interventions in Aging's Future Americans Model (Aim 1), align workflow and nudge performance in the electronic health record to develop the best choice architecture (Aim 2), and conduct a randomized trial of minimally disruptive changes to the electronic health record to encourage the use of evidence-based therapies for insomnia in everyday clinical practice at 60 clinics in the U.S. (Aim 3). The trial will employ the following two nudge concepts: 1) Social Accountability Nudges that ask a clinician seeking to prescribe a Z-drug to either justify, in a brief free-text response, the reason for a guideline-discordant action or to cancel the order, and to precommit to a brief deprescribing plan whenever prescribing a Z-drug; 2) setting default days supply, pre-populated in the medication order, to a small number of pills. In an 18-month trial involving 60 clinics (449 clinicians), clinic-clusters will be randomized to receive a) social accountability nudges, b) defaults, c) both social accountability and defaults, or d) no intervention (education control). The primary outcome measure is the change in Z-drug days supply ordered per patient. Secondary outcomes include probability of starting a new patient on a Z-drug, probability of discontinuation among long-term users, and a physician perceptions survey. To aid in deprescribing Z-drugs, we will also provide bibliotherapy, access to insomnia, referrals for screening for related problems and visit assessments. Sample size calculations indicate an 88.5% (95% CI: 83.3, 92.6) power to detect a 6.7% decrease in pills ordered. This proposal will integrate behavioral economic approaches into implementation research to encourage evidence-based decisions involving the treatment of insomnia. It will improve the safety and efficacy of everyday clinical practice for people visiting their physician with sleep problems. It will also provide estimates of the benefits of treatment to future Americans.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY / ABSTRACT Unsuccessful regulation of affect and physiology (RAP) is central to many forms of psychopathology, including depression, one of the leading causes of disability worldwide. A core feature of major depressive disorder is persistent sadness, which often results from difficulty successfully regulating affect and physiology. Heterogeneity in the course of illness and response to treatment in depression is likely due to the presence of multiple phenotypes that confer risk via distinct mechanisms. Individuals with remitted depression (rMDD) appear to be less successful than controls on two distinct dimensions of regulation in everyday life: Perceived success – how successful explicit regulation is at improving affect; and Physiological success – the degree of parasympathetic augmentation following regulation attempts, primarily an implicit regulation process. Classifying individuals into subgroups (i.e., phenotypes) according to scores on each regulatory dimension can have clinical utility, providing distinct mechanism-based targets for remediation based on explicit and/or implicit regulation. Our approach involves three phases: First, we plan to characterize regulation phenotypes by screening young adults with rMDD (n=184) and healthy comparisons (n=68) based on our previously successful dimensional ambulatory assessment of RAP in everyday life. Second, participants will complete deep phenotyping in the lab to validate neural and physiological mechanisms underlying their phenotypic RAP classification. Third, we will examine the remediating effects of heart rate variability (HRV) biofeedback on proximal measures of RAP success in everyday life, particularly for subgroups who demonstrated unsuccessful RAP. By identifying and elucidating mechanisms of phenotypes based on ecologically-valid measures of RAP success, this study will help to parse the heterogeneity that exists within depression. Determining how HRV biofeedback impacts everyday regulation success for specific phenotypes can serve as an initial step toward guiding precision medicine. Thus, this project is highly responsive to NIMH’s strategic plan to identify when, how, and for whom to intervene.

NIH Research Projects · FY 2024 · 2023-07

PROJECT SUMMARY/ABSTRACT An estimated 3.5 million youth between the ages of 18 and 25 experience homelessness in the U.S. each year. Relative to their stably housed peers, youth experiencing homelessness (YEH) are vulnerable to a multitude of risks affecting their health and well-being, including substance abuse, mental health challenges, victimization, and death. YEH are also more likely to engage in risky sexual behaviors and are up to 10 times more likely to contract HIV compared to their housed peers. Key to understanding the HIV risk and prevention behaviors of YEH are their social networks—their web of personal social interactions and relationships. Increased HIV risk among YEH has been tied to involvement in social networks with risk-taking peers and predominantly the consequence of unsafe sex over injection drug use and needle sharing. Conversely, connections to family and pro-social peers have been demonstrated to influence HIV prevention behaviors among YEH, including condom use and HIV testing. However, the effect of different living situations among YEH—and particularly of precarious housing (i.e., “couch-surfing” or temporarily staying with others)—on HIV risk and prevention behaviors remains largely unknown. A sizable portion of the YEH population reports living in precarious housing, and current evidence also suggests that that youth of color and youth identifying as LGBTQ+ are more likely to be precariously housed compared to other YEH, populations that also carry disproportionate risk for HIV. The proposed mixed-methods study has three aims. Aims 1 and 2 will use an existing dataset from Have You Heard?, a longitudinal study of a peer leader training intervention for HIV prevention among YEH in Los Angeles, California. Aim 1 will entail conducting an egocentric social network analysis to investigate whether precarious housing and social support are associated with HIV risk (e.g., transactional sex, condomless sex, sex under the influence, concurrent sex partners) and prevention (e.g., recent HIV testing and PrEP awareness and use) behaviors among YEH. Aim 2 will use latent class analysis to explore the heterogeneity of YEH based on living situation and social support networks in order to examine (a) whether LGBTQ+ identity, racial/ethnic minority status, and duration of homelessness relate to emergent subgroups and (b) whether HIV risk and prevention behaviors vary across these emergent subgroups. Aim 3 will entail conducting qualitative interviews with precariously housed youth (n = 20) to explore how social support networks and housing status influence HIV risk and prevention behaviors among precariously housed YEH. This study will expand knowledge regarding HIV risk and prevention behaviors among YEH, paving the way for the development of more tailored HIV prevention interventions for this vulnerable population. In addition, the PI will receive training in understanding HIV risk and prevention among vulnerable youth populations, advanced statistical methods, and mixed-methods research; gain experience conducting HIV-related research with YEH; and opportunities for professional development.

NIH Research Projects · FY 2025 · 2023-07

Abstract Spontaneous bacterial peritonitis (SBP) is a major complication of cirrhosis and associated with high mortality rate. Moreover, the effectiveness of antimicrobial therapy, the current standard of care, is deteriorating due to the global emergence of multidrug-resistant pathogens. Hence, it is essential to develop novel management strategies, for which furthering our understanding of the pathophysiology is imperative. Gut dysbiosis and impaired intestinal immunity in cirrhosis cooperatively predispose to the translocation of gut flora into the mesenteric lymphatics and, subsequently, to the peritoneum. Although this mechanism has been regarded as the primary pathophysiology, SBP develops only when the balance between peritoneal antibacterial immunity and bacterial virulence shifts in favor of the invading pathogen. Peritoneal macrophage (PM) is the major cell type in the peritoneum and is considered as the first line of antibacterial defense owing to the robust capacity in the engulfment and phagolysosomal digestion of pathogens. Moreover, PM sensing of microbe results in the production of cytokines/chemokines required for neutrophil recruitment and activation. Accordingly, PM is also essential for triggering the second wave of antibacterial response. Despite its perceived importance, the role of PM in SBP has been poorly understood. Furthermore, it remains largely elusive whether the development of cirrhosis alters the antibacterial properties of PM, and how it impacts the overall potency of peritoneal antibacterial immunity. Our single cell RNA sequencing analyses of peritoneal immune cells of rat with cirrhosis demonstrate a profound alteration of PM characteristics, including a substantial downregulation of the master regulator of PM, GATA6, a vitamin A (VA)-inducible transcription factor. In addition, PM of cirrhotic rats exhibit a markedly reduced capacity to produce inflammatory mediators in response to bacterial peritonitis, resulting in impaired neutrophil recruitment and activation as well as the insufficient bacterial clearance. Our studies also reveal that two common features of cirrhosis, VA deficiency and ascites accumulation, both independently contribute to the dysregulation of PM antibacterial function, with both processes involving GATA6 downregulation. Moreover, PM isolated from cirrhosis patients demonstrate a decreased number of GATA6-expressing PM, which correlates with the degree of antibacterial function impairment. These findings led to our hypothesis: “cirrhosis impairs the antibacterial properties of PM through the downregulation of GATA6 expression, which contributes to SBP development”. Accordingly, this proposal aims to close the knowledge gap regarding the role of PM in SBP through defining: (Aim 1) the impact of cirrhosis on PM antibacterial functions, (Aim 2) the influence of VA insufficiency on PM antibacterial properties, and (Aim 3) the effect of cirrhotic ascites on the antimicrobial activity of PM. The successful completion of the proposed studies will result in a paradigm shift in our understanding of SBP pathophysiology and ultimately define therapeutic targets for the development of novel therapeutic strategies.

NIH Research Projects · FY 2025 · 2023-07

Abstract The lymphatic system is the central regulator of fluid homeostasis in the body through absorbing and returning interstitial fluid back to blood circulation; additionally, it plays pivotal roles in lipid digestion and immune and inflammatory response through the lymphoid organs and immune cell trafficking. Studies have documented the presence of lymphatics in the uterus; however, little consensus exists regarding the exact development and distribution of the uterine lymphatics nor their impact on uterine health functions. Understanding the proper development and maintenance of the lymphatics may be of clinical importance, as lymphatics have been implicated in pathogenesis uterine diseases such as endometriosis and cancer metastasis. In this study, we propose to elucidate the development and functions of uterine lymphatics using Prox1 fluorescent lymphatic reporter and lymphatic loss of function mouse models. Based on preliminary data, we hypothesize that the uterine lymphatics develop postnatally under patterning guidance from uterine blood vessels, and that uterine lymphatics remodel to maintain fluid homeostasis of the uterine environment through estrous phases and pregnancy. After characterizing normal development of the murine uterine lymphatics, we will determine the function of uterine lymphatics in the context of fluid drainage of the uterine tissue and regulation of embryonic implantation sites and gestation. The maintenance of fluid homeostasis in the uterus through estrous phases and pregnancy will be assessed through real-time tracer studies and wet/dry tissue weight ratios, while the role of uterine lymphatics in embryonic implantation will be dissected through use of novel genetic and surgical inducible lymphatic loss of function mouse models. Together, these study outcomes will generate a novel understanding of murine uterine lymphatic morphology and function, which may provide a valuable new viewpoint from which to assess uterine health.

NIH Research Projects · FY 2024 · 2023-07

Optical Coherence Tomography (OCT) has profoundly impacted diagnostic imaging of the human retina, enabling accurate and early diagnosis of retinal disease. The relatively high cost of current clinical instruments limits its use in cost sensitive environments, e.g. screening in the optometrist's or general practitioner's office, underserved or rural US populations and developing countries. Motion of the patient and/or interferometer strongly effects phase stability and image quality, impacting the common approaches which scan a focused beam across the retinal surface. Collecting the entire field at once via an approach such as Full-Field Optical Coherence Tomography (FF-OCT), would obviate this issue and provide high spatial phase stability. Here we propose a novel solution that not only overcomes these issues (poor phase stability and coherent artifact) but promises to do so while reducing costs and increasing imaging speed. We will take advantage of rapid development in two key technologies, additive manufacturing for optical components and extremely high megapixel CMOS arrays. Building on concepts from our prior work, we will develop an FF- OCT system by mapping the 3-dimensions (x,y, and k) onto a 2-D CMOS array within an imaging spectrometer. This will in turn allow us to utilize inexpensive spatially incoherent light sources which do not produce coherence artifacts, but heretofore were limited to use with time-domain FF-OCT systems. This will be accomplished via 3 Specific Aims. Aim 1, Develop a 3-D printed structure for image remapping in FF-OCT: We will deploy a micro- optical design consisting of arrays of 3-D printed single mode fibers that map x,y in the image plane, to spaced columns. The space between the columns will enable dispersion in k within the 2-D spectrometer developed in Aim 2. This structure will enable volumetric OCT imaging with a single camera exposure by mapping x, y, and k onto a 2-D array. Aim 2, Develop an imaging spectrometer and spectral domain FF-OCT system around a high megapixel (Mp) CMOS sensor: We will design and develop an imaging spectrometer that will be compatible with the structure(s) from aim 1, integrating the structures as they become available. Aim 3, System validation and human retinal imaging: After validation on tissue phantoms, pilot human data from a spectrum of retinal diseases will be acquired and compared with standard-of-care clinical imaging systems. We expect the completed system to have an FOV of 5 mm diameter, 15.5 µm lateral sampling, 7.5 µm axial resolution and be able to collect a volume image in as little as 200 µs at 48 volumes per second.

NIH Research Projects · FY 2025 · 2023-07

The exon-1 of mutant huntingtin protein (mHTTex1) accumulates in the brains of Huntington’s disease (HD) patients and is implicated in neurodegeneration. The intrinsically disordered mHTTex1 misfolds into a heterogeneous mixture of assemblies, however, the pathogenic conformers are not well characterized. The major limiting factors have been the lack of methods to assemble ultrapure mHTTex1 structures, molecular tools to identify them and models to investigate their neurotoxicity. Towards this end, we have developed protocols to assemble distinct oligomers, protofibrils and fibrils of mHTTex1 and have generated libraries of monoclonal antibodies (mAbs) to defined structures. We plan to characterize the binding of representative mAbs to conformations in various assemblies using biophysical and biochemical methods, explore whether the interaction of each mAb with its epitope affects the misfolding, seeding and aggregation in vitro, and examine the structures of mHTTex1 species upon binding to selected mAbs (aim 1). We have further developed a diagnostic platform to study the entry of structurally known mHTTex1 species into human neurons and their aggregation into neurotoxic assemblies. With this model, we plan to identify and characterize the neuroinvasive/neurotoxic species of mHTTex1, map their pathogenic conformations and determine their structures with biophysical methods including Cryo-EM. Moreover, we plan to discover the neuronal receptors, which participate in the entry of mHTTex1 and identify the interacting proteins, which are incorporated in the neurotoxic aggregates. In addition, we plan to examine for the presence of neuroinvasive mHTT species in the brains of HD patients and in human neuronal and mouse models of HD to validate the physiological relevance of the in vitro-assembled structures and any links to disease severity. These experiments may for the first time identify the structures of neurotoxic mHTTex1 at high resolution, a novel pathway for their production, and may provide targets for therapy development (aim 2). In aim 3, we will investigate the role extracellular mHTT in disease in the CNS of HD mice. In one set of experiments, we plan to inject neuroinvasive species of mHTTex1 into the brains of asymptomatic R6/2Q51 HD mice (express human mHTTex1 with 51Qs) and investigate their ability to enter neurons, trigger assembly formation and accelerate disease progression. Moreover, we will determine whether blocking any of the pathogenic conformations of mHTTex1 by AAV- mediated delivery of recombinant antibodies, which are secreted in the CNS, inhibits the entry, amplification and neurotoxicity of the injected species. Finally, given that mHTT is present in the CSF and plasma of HD patients and mouse models, we plan to investigate the therapeutic impacts of secreted recombinant antibodies on the accumulation of pathogenic mHTT assemblies and progression of HD-like pathology in the Q140 HD mice expressing full-length mHTT. These studies will fill some of the knowledge gaps on the role of extracellular mHTT in the pathogenesis of HD and may provide therapeutic targets and reagents.

NIH Research Projects · FY 2025 · 2023-07

Abstract Title: Targeting IKKepsilon-mediated nucleotide synthesis in KSHV-associated lymphoma Human gamma herpesviruses, including Kaposi's sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV), are causative agents of diverse malignancies in immune- compromised individual, including AIDS patients and organ transplant recipients. In addition to KS, KSHV is invariably associated with two types of lymphoma, primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). No vaccine or effective treatment is available for KSHV-associated malignancies, though antiviral therapy targeting viral thymidine kinase is an option with limited efficacy. We have an outstanding interest in virus-host interaction involving innate immune defense system. Recently, we discovered that KSHV exploits the IKKepsilon kinase to reprogram metabolism in KSHV latently-infected PEL cells. Specifically, KSHV activates IKKepsilon to fuel de novo nucleotide synthesis via activating key metabolic enzymes known as glutamine amidotransferases. In doing so, IKKepsilon promotes the proliferation of KSHV-infected PEL cells and depletion of IKKepsilon arrests these cells at G0/G1 phase. This study will delineate the molecular interaction that KSHV activates IKKepsilon in metabolic reprogramming to support immortal proliferation of PEL cells. We have developed novel small-molecule inhibitors of IKKepsilon and glutamine amidotransferases. We will explore these drug-like molecules to target IKKepsilon and glutamine amidotransferase to impede PEL cell proliferation. Our work will not only elucidate fundamental mechanism governing PEL cell metabolism and proliferation, but also provide proof-of-concept that targets host factors to treat KSHV-associated malignancies.

NIH Research Projects · FY 2024 · 2023-07

Project Summary/Abstract Oxytocin is an evolutionarily conserved neuropeptide primarily produced and secreted from the paraventricular and supraoptic nuclei of the hypothalamus. Widespread expression of the oxytocin receptor (OXTR) across various brain regions allows oxytocin to mediate a wide range of effects on behavior. Though classically known for its functions in mediating social and maternal behavior, a growing body of evidence from both animal models and human studies indicates that the neuropeptide oxytocin is a potent anorexigenic signal and a promising target for obesity pharmacotherapy development [1-4]. Indeed, intranasal oxytocin for weight loss is currently under investigation in an active clinical trial [5]. The interest in the oxytocin system as a target for obesity treatment is based, in part, on findings showing that intranasal oxytocin reduces intake of palatable foods in humans [6], and that these effects are more potent in participants with obesity [7]. A deeper understanding of the neural pathways and behavioral processes mediating oxytocin's effects on food intake and food-motivated behavior is imperative given the clinical potential for this system. Oxytocin has been most widely studied for its influence on social behavior. Food intake regulation is heavily influenced by social factors in both humans and rodents, yet the extent that oxytocin's influence on these two related fundamental behaviors is interconnected has not been previously investigated. Further, many of the behaviors modulated by oxytocin show sexual dimorphism, including both social behaviors and food intake control [8, 9]. Here we focus on the dorsal hippocampus (HPCd) as a candidate brain region where these functions overlap as HPCd oxytocin signaling facilitates social memory in sexually dimorphic ways [10] and this region has been recently linked to food intake control [11]. Our preliminary results show that HPCd oxytocin delivery in male rats reduces intake when isolated in the home cage, yet increases food intake in the presence of a familiar, but not an unfamiliar conspecific. Aim 1 experiments build off these findings using a novel social eating paradigm and both pharmacological and virogenetic approaches in males and females to investigate how HPCd OXTR signaling modulates food intake in social context- and sex-specific ways. Our additional preliminary data reveal that central oxytocin enhances social transmission of food preference (STFP) learning in a conspecific familiarity-dependent manner. A recent paper identified a role for the HPCd in mediating STFP learning through dopamine signaling [12]. Aim 2 experiments will build off these findings to investigate whether the HPCd is a site of action mediating oxytocin's effects on STFP (Aim 2), and will utilize pharmacological, neuroanatomical, and in vivo imaging strategy to assess the extent to which oxytocin and dopamine systems interact to regulate eating in a social context- and sex-dependent manner (Aim 3). Collectively, these experiments will reveal novel neural, physiological, and behavioral mechanisms mediating oxytocin's effects on food intake control.

NIH Research Projects · FY 2026 · 2023-07

The ɛ4 allele of apolipoprotein E (APOE4) is associated with accelerated aging and mortality as well increased vulnerability to Alzheimer’s disease (AD). Although the causal links between APOE4, aging, and AD risk remain to be fully defined, candidate mechanisms include regulation of energy metabolism and systemic and neural inflammatory tone. Our recent findings demonstrate that nutritional approaches, including fasting mimicking diet (FMD), exhibit anti-aging and protective properties across several age-related conditions. Indeed, pathways identified to be beneficially regulated by FMD and related nutritional interventions overlap with pathways thought to underlie relationships among APOE4, aging, and AD. In this project, we investigate the central hypothesis that the dietary intervention FMD will protect against APOE4 phenotypes that drive age-related cognitive impairment and AD pathogenesis. We will investigate this hypothesis by studying systemic and neural effects of FMD and related dietary interventions across age in mice with human APOE genotypes both in the absence and presence of AD transgenes. Our studies will interrogate several potential mechanisms hypothesized to underlie gene X environment relationships between APOE and diet with emphases on brain energy metabolism and regulation of microglial activation profiles. We propose three aims. Aim 1: Do dietary interventions protects against APOE4-associated aging phenotypes? We test the hypothesis that nutritional interventions will improve systemic and neural outcomes in the contexts of aging and APOE4 genotype. Studies will test the ability of diet to prevent vs treat APOE4-associated phenotypes. We will test the efficacy of low methionine or high ketone diets assess the hypothesized abilities of a fasting-mimicking mitochondrial-derived peptide to mirror protective actions of FMB across APOE genotype. Aim 2: Do dietary interventions provide protection against Alzheimer pathology in an APOE-dependent manner? The second aim is conceptually parallel to Aim 1 but with a focus on diet and APOE genotype in the context of AD pathology. These studies will be conducted in male and female APOE3 vs APOE4 AD mice to determine potential sex differences and at two different ages to consider efficacies for both the prevention and treatment. Aim 3: Do dietary interventions improve glial phenotypes associated with again, APOE4, and AD? We investigate the hypothesis that the primary mechanism by which dietary interventions protect against APOE4 phenotypes is improvement in glial transcriptomic profiles and functions. We will interrogate the hypothesis that a key regulator of APOE-dependent differences is the TREM2-ApoE signaling pathway, which is linked to AD risk via actions on microglial phenotype and metabolic fitness. Completion of the proposed studies will yield preclinical data that define the interactions and underlying mechanisms among diet, APOE genotype, sex, age, and intervention timing, relationships that are essential to the effective clinical translation of dietary strategies to combat cognitive decline and AD.

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY Preclinical and clinical data show elevated glutamatergic transmission and neuronal activity within the hippocampus during the early stages of schizophrenia development, and it is proposed that such elevations drive the progression of the disorder. However, the molecular and cellular mechanisms that give rise to schizophrenia- related elevations in hippocampal glutamatergic transmission are unknown. Recent human genetic data supports a connection between reduced DLG1 gene expression/function and schizophrenia. The DLG1 gene encodes the putative synaptic regulatory protein SAP97. However, a clear synaptic regulatory role for SAP97 has remained elusive. We have now, for the first time, identified a native synapse in the brain where SAP97 plays a critical and direct role in synaptic regulation. The long-term goal of our research is to understand how glutamatergic synapse dysfunction contributes to the development of psychiatric disorders. Our central hypothesis is that disruption of SAP97 function in dentate granule neurons (DG neurons) of the hippocampus produces pathological synaptic strengthening that contributes to schizophrenia etiology. Guided by strong preliminary data, we will pursue this hypothesis in three specific aims. In Aim 1, we will combine electrophysiological methods with brand new imaging tools to characterize pathway-specific roles of SAP97 in regulating glutamatergic neurotransmission and plasticity. In Aim 2, we will use a combination of newly generated antibodies, molecular biological and electrophysiological/imaging approaches to identify the roles of SAP97 subregions in governing SAP97’s subcellular localization and influence over glutamatergic synapse function in DG neurons. In Aim 3, we will deploy cutting-edge in vivo imaging tools in freely behaving animals in order to understand how hippocampal neuron activity is altered in rodents with compromised SAP97 function. The present proposal is innovative because it deploys new and powerful genetic, in vivo imaging, and behavioral analysis tools that will allow a precise characterization of a newly discovered role for SAP97 in regulating the function of native glutamatergic synapses in the hippocampus. The proposal is significant because it stands to identify synaptic pathology in a specific brain region that represents a common component in schizophrenia development. This proposal squarely meets the mission objectives of the NINDS given its focus on understanding glutamatergic synapse regulation and memory formation which are found to be disrupted in complex brain disorders.

NIH Research Projects · FY 2025 · 2023-06

Obesity is a major public health concern and is linked to increased risk for many other health outcomes including cancer, hypertension, type 2 diabetes, stroke, asthma, and early mortality. Housing quality and the built environment are believed to play an important role in shaping obesity and related health outcomes. However, it is unclear to what extent obesity can be reduced by improving housing and built environments, for example, by improving walkability, green space, healthy food access, safety, and air quality. To address this research gap, we propose to study a time-sensitive natural experiment that will ultimately significantly improve opportunities for health by altering the built environment and housing quality. The Housing Authority of the City of Los Angeles is planning to redevelop the Rancho San Pedro (hereafter, Rancho) community, an obsolete 478-unit public housing project located next to the Port of Los Angeles. The redevelopment will build new housing for existing residents, add additional housing units and homeownership units, provide several opportunities for healthier lifestyles via parks, green spaces, jogging trails, community gardens, street lighting, and retail space, and create housing and built environment enhancements that will reduce air pollution exposure from the Port. Current Rancho residents will continue to live in existing housing during the redevelopment, and as a result, residents will experience construction-related disruption (loss of greenspace and walkability, noise and air pollution). Our goal is to study the effects of these short-term disruptions on residents’ BMI and related health outcomes (e.g., cardiovascular, asthma exacerbation). We will do this by following a cohort of residents of Rancho and a control public housing site. In Aim 1, we will examine the short-term impact of the Rancho public housing redevelopment on residents’ BMI and related health outcomes. In Aim 2, we will unpack the potential mechanisms for the observed change in health outcomes to investigate both why and how observed changes occur. Finally, in Aim 3, we will explore moderating effects by time activity patterns and health status to understand for whom and under what circumstances changes were observed.

NIH Research Projects · FY 2026 · 2023-06

ABSTRACT Major depressive disorder (MDD) is the leading cause of disability worldwide, and around half of MDD patients have treatment-resistant depression. The use and clinical benefit of rTMS have escalated greatly in recent years. As only 40-50% of patients respond to current standard rTMS, there is great interest in predicting which patients are likely to respond, what brain features best predict response, and how these features relate to the core biosignatures of MDD. To address this, and responding to NIH’s call for Precision Medicine approaches, our Global Deep Learning Initiative to Understand Outcomes in Major Depression unites international leaders in MDD and rTMS research, neuroimaging, and AI to identify generalizable predictors of rTMS response, and assess how they relate to brain biomarkers of MDD. Two major innovations are proposed. First, we use novel deep learning methods, based on convolutional neural networks, to extract predictive features from multimodal brain images (sMRI, DTI, and rsfMRI); tactics applied in whole-brain and surface-based mapping of brain function and structure, DVAEs for feature extraction, and transfer learning (to learn from auxiliary datasets and tasks) will distill predictive features while protecting individual privacy. CNNs trained on multimodal brain maps for our predictive tasks will distill additional layers of information that have not yet been fully exploited in MDD research, to better predict clinical status and treatment response. Second, our worldwide ENIGMA-MDD network will provide diverse test data from globally representative populations, to ensure that our predictive models do not break down when tested on diverse data. ENIGMA’s harmonized extraction of brain measures across worldwide cohorts will enhance rigor and ensure that analyses are well-powered and consistently performed. We include an important partnership with REST-meta-MDD, a Chinese consortium collecting multimodal imaging data from patients with MDD, to test the generalizability of our predictive models. The likely outcome of our work is a set of pre-screening tools to predict who will respond best to rTMS, and a deeper understanding of the brain signatures of MDD that predict treatment outcomes following rTMS. All tools will be made public via NITRC and ENIGMA websites, and will be tested across our ENIGMA network, guaranteeing impact of the work for large- scale outcome prediction within and outside of MDD research.

NIH Research Projects · FY 2026 · 2023-06

Medulloblastoma (MB), is the most common pediatric brain tumor originating in the GABA-rich cerebellum. 40% of MB patients present as both cerebellar and cerebral spinal fluid (CSF) leptomeningeal metastases which results in having dismal outcomes. The project’s long-term goal is to determine how MB become life-threatening leptomeningeal metastases by studying the tumor and brain microenvironment from a neurodevelopment and cancer neuroscience perspective. Although abnormal GABAergic receptor activation has been described in group 3 MB, no studies until ours recently had yet to elucidate the contribution of receptor-independent GABA metabolism to MB pathogenesis and metastasis. Overall, we were the first to identify GABA metabolic shunt enzyme GABA Transaminase (ABAT) expression is correlated with cerebellar development and is used by MB to survive in the cerebrospinal fluid microenvironment and promote leptomeningeal dissemination. Specifically, we show proliferative MB cells at the primary cerebellar site have low ABAT. However, a sub-population of ABAT positive MB cells at the primary cerebellar site display neuron-like characteristics (including metabolism) and are the seeds of MB metastasis. In addition to being a GABA metabolic enzyme, we determined that expression of ABAT induces epigenetic modification by significantly reducing histone acetylation at the 4th lysine residue of the histone H3 protein (H3K4ac) and significantly increase in histone deacetylase activity. Furthermore, ABAT expression fluctuates depending on metabolite changes in the tumor microenvironment, with nutrient-poor conditions upregulating ABAT expression. We found metastatic MB cells require ABAT to maintain viability in the metabolite-scarce CSF by using GABA through the GABA metabolic shunt as an energy source substitute, thereby facilitating leptomeningeal metastasis formation. Therefore, we hypothesize ABAT has a dual-biological role: lack of its expression during normal cerebellar development contributes to proliferation leading to group 3 medulloblastoma formation; While its overexpression after MB development results in tumor dormancy leading to survival benefit in disseminated cells in leptomeningeal metastasis. Utilizing foundations of neurobiology by using: a) human and mouse-derived neural stem cells, neurons and glial cells, b) unique strengths of patient-xenografts, c) BarTeL transgenic mice, d) novel small molecule ABAT inhibitor which crosses the blood-brain barrier, we will: 1st interrogate the role of ABAT in cerebellar neurodevelopment and Group 3 MB initiation, 2nd contribution of nuclear ABAT to transcription through histone acetylation modification in metastases, 3rd investigate ABAT as a potential therapeutic target for MB metastases. The current proposal will shed further light on understanding how metastatic MB cells develop and adapt to their neural niche opening avenues for novel therapeutic interventions for children that have this devastating disease.

NIH Research Projects · FY 2026 · 2023-05

Alternative splicing (AS) is a major mechanism that generates the vast transcriptome and proteome diversity from the limited genome. Spatial and temporal regulation of AS contributes to cell differentiation and lineage determination. Mutations that disrupt splicing cause a wide range of diseases including neurodegeneration, muscular dystrophies, and cancer. Therefore, understanding and targeting splicing is essential to the future of precision medicine. In the past 3 decades, AS studies have focused on cis elements and their associated trans factors, such as RNA binding proteins (RBPs). The laws of thermodynamics dictate that RNAs fold into low free energy structures in the context of RNA-protein complexes (RNPs), however, very little is known about how pre-mRNA structures in large RNPs control splicing, and various RNA processing events in general. Our lab invented several chemical crosslink-ligation based methods that enabled direct analysis of transcriptome-wide protein-independent RNA 2D and 3D structures in vivo (PARIS and SHARC). In this proposal, in Aim 1, we develop and benchmark a high throughput technology, SHARCLIP, to directly capture all RNA structures, RNA-RNA, RNA-protein, and protein-protein interactions together. SHARCLIP is conceptually similar to methods that analyze protein-mediated chromatin conformations, e.g., ChIA-PET and hiChIP, and will bring 1D RNP interaction studies to higher dimensions. In aim 2, applying PARIS and SHARC to chromatin associated RNAs, and the new method SHARCLIP to key splicing regulators, we will build transcriptome-wide RNA-structure and RNA-protein interaction models and deconvolve their dynamics in 2 ENCODE cell lines HepG2 and K562, as well as 6 brain cell lineages from a human iPS differentiation system. In aim 3, further integrating cell type specific AS programs and disease variants implicated in splicing, we will test whether mutations act by altering the structures. With the structure models as a guide, we will use a combination of CRISPR genome editing and structure-perturbing antisense oligos to test the roles of specific structures in regulating splicing in neuronal differentiation. Together, this proposal will produce a new technology that simultaneously capture multi-valent RNA-protein interactome and RNA structurome in vivo, establish a structure-based splicing code, provide an important resource to enable mechanistic studies of RNA processing, and pave the way for future therapeutic targeting of splicing.

NIH Research Projects · FY 2026 · 2023-05

Project Summary/Abstract For this application, “Studying the Genetics of Aging, Behavioral, and Social Phenotypes in Diverse Populations,” we propose to develop tools to promote genetic research of aging, behavioral, and social phenotypes in diverse populations. These phenotypes have a number of unique characteristics (e.g., polygenicity, environmental mechanisms, and small effect sizes) which require special consideration when developing research tools. In brief, we propose to: • Develop the Genetic-Related-Matrix-Matched Association study (GRMMA) tool for performing genome-wide association studies (GWASs) in large, diverse data sets. Current GWAS methods require restricting samples into approximately homogeneous-ancestry samples, which is wasteful and has resulted in Eurocentric bias in genetics research. Using matching methods, GRMMA can use more of the available data in a way that both reduces bias and increases statistical power. We will employ computationally efficient strategies that allow us to implement GRMMA in large diverse sample such as the UK Biobank. We will make the GRMMA tool and tutorials publicly available through the online repository, Github. • Develop SBayes-Universal (SBayesU), an efficient new tool for producing polygenic scores (PGSs) by optimally combining GWAS summary statistics estimated in different populations. The key feature of SBayesU is that it uses a low-dimensional eigen decomposition of the linkage disequilibrium matrix. This permits SBayesU to model a much larger set of SNPs, to model SNP annotations, to account for imperfect cross-ancestry genetic correlation, to produce PGSs for populations that are not included among the sets of GWAS summary statistics, and to allow our algorithms to converge much more quickly and reliably. We will also make the SBayesU tool and tutorials publicly available. • We will apply the best available method for producing diverse-population PGSs (which we anticipate will be SBayesU) to a wide range of aging, behavioral, and social phenotypes, using existing cohorts and new genotyped data that becomes available during the grant period. We will make the polygenic scores we produce publicly available as part of the Social Science Genetic Association Consortium’s Polygenic Index Repository, which currently creates polygenic scores for 11 widely used datasets (but currently only for the European-ancestry individuals in those datasets). Each release of the Repository will be accompanied by documentation that clearly describes methods used and the underlying data.

NIH Research Projects · FY 2025 · 2023-05

Project Summary/Abstract More than 52 million American adults report binge drinking in the past month. Binge drinking is intricately linked to, and exponentially increases the risk for, the development of an alcohol use disorder - a pernicious illness linked to severe medical and psychiatric morbidity, elevated mortality, and a resistance to conventional treatments. In tandem, up to 43 million Americans engage in episodes of binge eating, characterized by the consumption of a large amount of food coupled with a loss of control. Binge eating is inherently linked to the development of binge eating disorder, the most prevalent eating disorder phenotype, and obesity, which is among the leading causes of global preventable death. Crucially, binge drinking and binge eating frequently co-occur, with robust data illustrating a bidirectional antagonistic effect of one binge type behavior upon the severity and frequency of the other, indicative of shared underlying mechanisms. Although the shared mechanisms underpinning both binge drinking and binge eating have been well-explicated, few treatments to date have been developed to target these transdiagnostic maintaining factors. Addressing these mechanisms early through the use of accessible prevention approaches has important public health implications for preventing the development of harmful and difficult-to-treat disorders. Importantly, the risk for both binge drinking and binge eating are substantially elevated in young adults, and particularly among college students, where paradoxically, rates of treatment seeking for these problematic behaviors are reliably low. Since both alcohol use disorder and binge eating disorder most typically onset in late adolescence, precision methods to target these symptomatic behaviors prior to their conversion to full threshold disorders, in the populations most at-risk, is of critical importance. Moreover, and owing to the reliably low rates of treatment seeking in college students, the development of precision approaches that are personalized and accessible on a large scale are especially warranted. In this study, we propose to leverage our group’s experience in developing mobile phone-based behavioral health interventions, to develop a mobile intervention to dually reduce co-occurring binge drinking and binge eating by targeting their shared underlying mechanisms. This two-phase study will first seek to develop and conduct a systematic beta test of the intervention, comprised of components of evidence-based treatments vetted with experts, in 20 male and female college students who engage in binge drinking and eating. Subsequently, we will pilot test the refined intervention in a randomized controlled trial of 300 college students who engage in both behaviors, and who will receive the intervention (N=150) or standard receipt of resources (control; N=150), to ascertain intervention efficacy in reducing binge frequency and related problems. Results will provide the first known data relating to a scalable and transdiagnostic approach to reducing binge drinking and eating frequency in a population at heightened risk and limited treatment-seeking.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY Alcohol misuse is rising in females. As females may experience worse health consequences then males and female-specific risks associated with alcohol misuse, elucidating female-specific factors is essential to build a better understanding of alcohol misuse in females and the development of prevention strategies for alcohol misuse in females. A small literature implicates dimensional premenstrual dysphoric disorder (dPMDD)—a mood disorder characterized by clinically significant emotional, behavioral, and physical symptoms during the late luteal that completely resolve by the mid follicular—as a critical female-specific risk factor for alcohol misuse and luteal phase increases in alcohol use. However, no studies to date have evaluated the shared physiological and emotional pathways of dPMDD and alcohol misuse. Emotion regulation encompasses the awareness and identification of emotion, and strategies to modify the emotional response. Emotion regulation may also be physiologically represented. Heart rate variability (HRV), an index of parasympathetic control over heartrate, is considered a peripheral psychophysiological marker of emotion regulation. Reduced subjective and objective (i.e., HRV) emotion regulation has been shown to associate with alcohol misuse. However, studies have not investigated how cyclical changes in emotion regulation may act as a risk factor for alcohol misuse and whether this differs between females with compared to without dPMDD. HRV shows the lowest levels during the luteal phase and our pilot data shows greater luteal phase reductions in HRV map onto more significant increases in luteal phase negative emotion. Our central hypothesis is that females with dPMDD suffer from recurrent luteal phase increases in heavy drinking due to progesterone-related declines in HRV and associated deficits in emotion regulation. The proposed study will recruit premenopausal females with heavy alcohol use (50% dPMDD) who will complete 4 weeks of prospective screening; and 5 weeks of Ecological Momentary Assessment (EMA) and HRV assessment. Aim 1 tests the hypothesis that dPMDD will show larger luteal phase increases in alcohol misuse relative to no dPMDD. Aim 2 tests the hypothesis that cyclical deficits in emotion regulation will predict cyclical increases in alcohol misuse. Aim 3 tests the hypothesis that individual differences in the degree of HRV reduction from the follicular to the luteal phase will predict greater luteal phase increases in alcohol variables. The results of this study will importantly contribute to the NIH strategic plan for research addressing sex/gender differences and female health. Specifically, this study will provide information on cyclical emotion regulation associations with alcohol misuse, which will provide foundational information regarding the influence of emotion regulation in the context of heavy drinking and the etiologic overlap between female alcohol misuse and emotion regulation in dPMDD.

NIH Research Projects · FY 2024 · 2023-05

Summary: Brain capillary Piezo1 ion channels and blood flow regulation in Alzheimer’s Disease Brain capillaries are the smallest blood vessels in the brain and are crucial in sensing the metabolic needs of active neurons and responding to these needs by supplying more blood. Notably, this function is impaired during neurodegenerative diseases such as Alzheimer’s disease. Not only that capillary sensing is impaired during Alzheimer’s disease, vascular stiffness and complete disruption of capillary blood flow, referred to as capillary stalling, have been observed in mouse models of Alzheimer’s disease. Given that capillary stalling disrupts the hemodynamic forces that capillaries are exposed to, it is important to understand whether Alzheimer’s disease associates with altered mechano-sensing abilities. We recently discovered that the protein Piezo1 is a mechanosensor in brain capillaries. Here, we test the overall hypothesis that altered Piezo1 channel activity is involved in the reduction of brain blood flow in Alzheimer’s disease. We further speculate that tuning Piezo1 activity will improve cerebral blood flow. We will investigate the contribution of endothelial Piezo1 channels to capillary stalling and blood flow reductions in a mouse model of Alzheimer’s disease. Using capillary endothelial cells from wild-type and Alzheimer’s disease mice, we will employ innovative electrophysiological and molecular approaches to assess whether Piezo1 ion channel expression and function are altered during Alzheimer’s disease. In the second aim, we will utilize in vivo high-resolution, two-photon microscopy to investigate capillary stalling and blood flow changes in Alzheimer’s disease mice while pharmacologically manipulating Piezo1 channels. This novel work could lead to new therapeutic strategies not previously tested.

NIH Research Projects · FY 2025 · 2023-04

ABSTRACT. Per- and poly-fluoroalkyl substances (PFAS) are a class of persistent, ubiquitous, and endocrine disrupting synthetic chemicals commonly used in consumer products and industrial processes. While they are detected in >97% of the US population, residents of low-income communities and racial-ethnic minorities show greater concentrations of many PFAS substances. Experimental and animal data support PFAS as potential mammary carcinogens. Epidemiologic data have been mixed, but results from a limited number of prospective studies among primarily European white women suggest that several PFAS compounds are associated with increased risk of breast cancer and that metabolic alterations linked to PFAS might underlie the associations. However, no prospective data from diverse racial-ethnic populations are available. The primary goal of this study is to investigate whether higher PFAS exposure increases the risk of breast cancer in a diverse US population and whether integration of PFAS levels, metabolic alterations, genetic predisposition, and epidemiologic risk factors of breast cancer identifies subgroups of women at increased risk. We propose the first and largest prospective multi-ethnic study to examine PFAS as novel risk factors of breast cancer. Leveraging the pre- diagnostic blood samples and comprehensive data on breast cancer risk factors and genomics in the Multiethnic Cohort (MEC), an ethnically diverse prospective cohort in California and Hawaii, we propose a nested case- control study of 1,600 female invasive breast cancer patients and 1,600 matched control women (12% African American, 16% Latina, 11% Native Hawaiian, 40% Asian American, and 21% White). We will utilize state-of-the- art laboratory technologies based on untargeted liquid chromatography with high resolution mass spectrometry to measure plasma PFAS and metabolome profiles, and combine these comprehensive measurements with innovative statistical methods to evaluate the integrated associations of PFAS, metabolome, genetics, risk factors, and breast cancer risk. Our specific aims are: (1) To examine the association between pre-diagnostic plasma levels of PFAS and risk of invasive breast cancer; (1a) Evaluate the associations of individual PFAS and PFAS mixture in pre-diagnostic plasma samples with breast cancer risk; (1b) Examine whether genetic predisposition determined by a polygenic risk score (PRS) modifies the association of individual PFAS and the PFAS mixture with breast cancer risk; (2) To examine the association between pre-diagnostic metabolomic profiles (a) with PFAS exposures and (b) with invasive breast cancer risk; (3) To integrate PFAS concentrations, metabolomic measures, PRS, and other breast cancer risk factors to identify subgroups of women at increased risk for breast cancer using an innovative latent variable analysis. This research relies on a multidisciplinary team of internationally-recognized investigators with expertise in environmental and cancer epidemiology, and state- of-the-art high-resolution metabolomics, genomics and statistical methods. Findings have the potential to inform precision medicine approaches for breast cancer prevention for diverse populations, and to impact policy change.

NIH Research Projects · FY 2026 · 2023-04

Electrification of the transportation sector in the United States is projected to have considerable cobenefits for public health through reductions in tailpipe-related emission. Numerous studies have estimated impacts on air quality and a few on health outcomes under various hypothetical scenarios— but no study to date has evaluated the broad real-world impacts of current rapidly increasing electric vehicle (EV) adoption levels. Further, the is a need to understand the distribution of EV adoption to ensure all people are benefitting from this transition. We propose to study patterns of EV adoption and in the observed local air quality and respiratory health co-benefits of EV adoption, using a sequential qualitative  quantitative  qualitative mixed methods design which melds community-engaged research with traditional epidemiological data analyses of large statewide databases in CA. We will first conduct focus group discussions with community members of the Southeast Los Angeles region, in collaboration with our community partner, to better understand perceived barriers and possible opportunities for promoting adoption of EVs in traffic-burdened communities (Aim 1a). Data from these focus group discussions will be used to refine our preliminary conceptual framework for EV adoption and identify additional hypotheses for testing in subsequent quantitative aims. Next, key predictive factors of neighborhood-level EV adoption in CA will be characterized using real-world data on EV registration (Aim 1b). Then, we will evaluate the effect of observed local EV adoption on measured tailpipe-related air pollutant concentrations (fine particulate matter [PM2.5] and nitrogen dioxide [NO2]) and assess potential effect measure modification by neighborhood/contextual factors (Aim 2). Finally, we will examination associations of EV adoption with local ED visits and hospitalization for pediatric and adult asthma and COPD, again assessing for potential effect measure modification by neighborhood/contextual factors (Aim 3). Sustained bidirectional community engagement is built in across the entire study period. In addition to focus group discussions, the community engagement plan includes annual community dialogues and Advisory Council meetings and a final community report back meeting. Our multi-pronged approach for community engagement is expected to enhance the relevance, validity, and impact of research findings. Study findings will provide policymakers with valuable real-world evidence on air quality and health co-benefits of the EV transition and capacitate communities with high traffic pollution burden in the transition, facilitating future research to reduce future environment and health impacts.