University Of California-Irvine

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY Asian Americans have more sleep deficiencies compared to Whites or Hispanics. However, most studies of sleep disparities have focused on African Americans and Hispanics, and research on Asians remains scarce. Our preliminary study among Chinese, Korean, and Vietnamese Americans found that acculturative stress was inversely associated with sleep duration and positively associated with sleep disturbance and risk of sleep apnea. Importantly, COVID-19 has adversely affected health, including sleep, with minority populations being disproportionately affected. Additionally, increasing hate speech and racist attacks against Asians have been reported, leading to increased mental and emotional toll in this population. Findings from the preliminary study also demonstrated that sleep duration was inversely associated with diabetes, and sleep apnea was positively associated with hypertension and obesity. The overarching goal of this innovative longitudinal study is to understand: (1) mechanisms of sleep disparities in relation to immigrant stressors and protective factors; and (2) consequences of sleep disparities on health outcomes, in a sample of community-dwelling Chinese, Korean, and Vietnamese adults (n=750). This study includes several innovations such as novel measures on anti-Asian racism due to COVID-19, the adverse impact of COVID-19, and multi-dimensional sleep health; use of dried blood spots; purposive sampling of 3 Asian subgroups; and considering ethnic enclaves as a protective factor. Over the 5-year project period, the investigators will collect and analyze two waves of data in order to: (1) Determine the longitudinal association between immigrant stressors (e.g., acculturative stress, psychological stress and adverse impact of COVID-19, anti-Asian racism, and neighborhood disadvantage) and sleep health (a multi- dimensional assessment of sleep health and disturbance); (2) Evaluate the moderating effect of potential protective factors (e.g., social support, religious involvement, ethnic enclaves, and neighborhood social cohesion) on the associations between immigrant stressors and sleep health; (3) Examine the longitudinal association between sleep health and markers of cardiometabolic risk; and (4) Test whether specific dimensions of sleep health will mediate the association between immigrant stressors and health outcomes. This will be one of the first longitudinal studies to investigate mechanisms of sleep disparities in relation to immigrant stressors and the consequences of sleep disparities on health outcomes among Asians, an understudied minority population which displays poorer sleep outcomes relative to other groups. This study is timely considering the rapid growth of Asians in the U.S., and the current hostile environment for Asians and immigrants, including COVID-19. This innovative study will elucidate health issues of this understudied group and identify modifiable factors that will serve as targets for intervention to reduce sleep disparities among Asians.

NIH Research Projects · FY 2025 · 2021-04

Autism Spectrum Disorder (ASD) is a prevalent and heterogeneous neurodevelopmental disorder with high co- morbidity for intellectual disability. This includes difficulties forming episodic, personal narrative, memories that are critical for orderly thinking and organizing future behaviors. Episodic memory deficits are thus thought to be major contributors to cognitive difficulties associated with autism. Many brain changes underlying abnormalities in ASD appear in childhood suggesting the possibility for effective therapeutic strategies targeting brain maturation. One candidate therapeutic is the hypothalamic peptide Oxytocin (OXT). Postnatal OXT treatment improves social behavior in animal models of ASDs and recent work indicates that treatment in childhood improves social interactions in autistic individuals. OXT acutely facilitates forms of synaptic plasticity, but there has been little experimental consideration of possible enduring effects of postnatal OXT treatment on learning and no analyses of effects on episodic memory. We examined this possibility using intranasal OXT (iOXT) treatment in the Fmr1 KO mouse model of Fragile X Syndrome, and novel paradigms for analyses of `What, When and Where' encoding. Our preliminary results show that in Fmr1 KOs iOXT treatments during the second postnatal week (P7-13) fully rescue hippocampal field CA1 long-term potentiation, object location memory, object identity (What) learning, and social recognition as assessed in adulthood (i.e., >40d after the last treatment). These findings raise the exciting possibility that a limited period of early life OXT treatment can effect a life-long rescue of a critical element of cognitive function in ASD. They also raise questions as to the breadth of effects iOXT has on behavior and the mechanisms involved; these questions will be addressed in the proposed studies. Aim 1 studies will test if postnatal iOXT treatment of male and female Fmr1 KO mice rescues encoding for the three major components of episodic memory, social recognition and stereotypic behavior as assessed in adulthood, and if effects depend on native OXT efflux. We will also determine if there is a critical period for enduring iOXT effects on behavior. Aim 2 will use electrophysiological recordings of evoked responses and network activity, analyses of synaptic proteins and signaling, and measures of neuronal arbors to test if postnatal iOXT treatment normalizes neurobiological processes in the distinct hippocampal subdivisions related to episodic memory encoding. Finally, Aim 3 will test the hypothesis that early life iOXT leads to activation of synaptic trophic factor receptors (EGFR, TrkB) in hippocampus, thereby suggesting a direct route for OXT effects on maturational changes in the structure. Overall, the proposed studies will greatly expand our current knowledge of OXT actions in the young brain, including potentially critical roles in regulating hippocampal development and synaptic function. Moreover, the results will lay the groundwork for designing novel, early life regimens to optimize hippocampal maturation and function, and to rescue the encoding of episodic memories in ASD and related developmental disorders.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY Situational cues that signal reward availability can exert a powerful invigorating influence over reward-seeking behavior. However, this impulse to seek out reward is not always adaptive. To conserve time and effort, cue- motivated reward seeking is regulated by homeostatic and cognitive control processes. There is growing evidence that these processes become dysregulated in a range of neuropsychiatric diseases, including substance use disorder, compulsive overeating, and depression, leading to a drop in healthy reward-seeking activity (e.g., apathy and anhedonia) and/or the development of maladaptive reward seeking (e.g., intense drug and food cravings). Despite their importance to both health and disease, much remains unknown of the neural circuits that regulate adaptive cue-motivated behavior. This project aims to fill this critical gap in knowledge. Based on the recent studies and our strong preliminary findings, we hypothesize that dopamine release is transformed into a motivational message at nucleus accumbens (NAc) terminals by cholinergic activity, and that inputs from the paraventricular thalamus (PVT) and anterior cingulate cortex (ACC) exert opposing regulatory influences over this process to ensure adaptive reward seeking. We will rigorously test this using a multifaceted approach that combines projection- and cell type-specific activity monitoring, neurochemical recordings, projection- and cell type-specific chemogenetic and optogenetic manipulations, and a translationally relevant Pavlovian-to-instrumental transfer assay of cue-motivated behavior. Aim 1 will investigate how cholinergic modulation of NAc dopamine release contributes to homeostatic and cognitive control over cue-motivated reward seeking. We will specifically determine whether cue-elicited NAc dopamine encodes changes in need state and reward probability, how this relates to midbrain dopamine neuron activity, and whether dopamine’s motivational message is locally shaped by cholinergic interneurons acting at β2-containing nicotinic acetylcholine receptors on dopamine terminals. Aim 2 will investigate if projections from PVT to NAc facilitate cue-motivated behavior in line with current needs, and whether it does so by regulating NAc cholinergic and/or dopaminergic activity. Aim 3 will determine if ACC projections to NAc adaptively suppress active reward-seeking behavior when cues signal that an alternative response would be advantageous, and whether this depends on NAc acetylcholine and/or dopamine. Our findings will lead to major advances in knowledge of the specific neural circuits and neurochemical mechanisms responsible for regulating cue-motivated behavior, and will guide future research on how dysfunction in these mechanisms contributes to maladaptive reward seeking in addiction and related disease states.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY Sporadic inclusion body myositis (sIBM) is a rare disorder of aging Americans, causing asymmetric muscle weakness and severe disability and morbidity. It is currently untreatable, and poorly understood. The prevalence of sIBM is likely to increase as the proportion of the United States population above the age of 65 years continues to grow. A major barrier to clinical trials in sIBM has been the lack of full understanding of the natural history of the disease. It remains to be determined whether the rates of disease progression is uniform and whether the various biomarkers associated with sIBM (anti-NT5c1A antibodies, variant T-cell populations) influence the natural history and disease behavior. Given the slow rate of disease progression, such observations cannot be made in the context of a routine clinical trial, and such studies need to be done as a separate stand-alone observational study. To address these unmet needs, we propose a prospective study with four specific aims. Aim 1: To determine for the first time whether c1A antibodies mediate disease progression over a two year interval in patients with sIBM. Aim 2: To perform a detailed morphological, histochemical, and immunohistochemical analysis of fresh muscle biopsy specimens obtained from a subset of patients with sIBM. Aim 3: To characterize the distribution of “immunosenescent” lymphocytes in circulating blood from patients with sIBM. Aim 4: To quantify the decline in the respiratory function of sIBM patients. The significance of our proposed study is 1) to allow for a detailed characterization of the disease progression in sIBM over a two-year period, and 2) to explore the relationship of a number of biomarkers associated with sIBM, and their influence on disease behavior and disease progression. Upon completion of these aims, we will 1) understand the disease phenotype, including pattern of respiratory involvement, and disease progression in sIBM better and understand the influence of serum antibodies to NT5c1A antibodies on the natural history and disease behavior; 2) define differences in serum variant T-cells and cytokine signatures in sIBM patients and their influence on disease progression and behavior; and 3) understand muscle pathology and immune cell distribution in sIBM patients and its relationship to NT5c1A antibodies. These findings may influence future trial design in sIBM. Finally, we will have created a thirteen-site consortium of myositis treatment centers that will be ready to adopt quickly any future clinical trials aimed at changing the course of sIBM.

NIH Research Projects · FY 2026 · 2021-04

Project Summary / Abstract The sequencing of the human genome promised a user manual for proteome-wide drug discovery, and our laboratory has been prototyping next-generation library synthesis and screening technology to fulfill that promise. As genome sequencing gave way to high-throughput screening (HTS) through the NIH Molecular Libraries Program, a new problem emerged: druggability. Most human proteins drive function not through functional-group-rich enzymatic active sites that small molecules can easily inhibit, but through large, featureless surfaces of protein-protein interfaces. These targets require larger, bespoke molecules absent from the “drug-like” collections in HTS libraries. Encoded library technologies, which combine nucleic acid information storage with combinatorial chemical synthesis (e.g., DNA-encoded libraries or DEL and mRNA display), emerged as powerful tools to access and screen these bespoke chemical spaces. Both technologies yield large libraries (10⁹–10¹³) interrogated by affinity to identify binders to purified protein targets. However, drug discovery teams generally require hits that influence biological function in living cells, tissues, or animals. Our laboratory develops the scalable interface between affinity selection outputs (10³–10⁵ molecules) and the few tens of hit structures synthesized to initiate medicinal chemistry. We devise synthesis technologies to transform selection outputs into one-bead-one-compound (OBOC) encoded libraries, with each bead displaying many copies of one library member. These enable activity-based screens when beads are individually compartmentalized with assay reagents. We have developed biochemical microfluidic OBOC-DEL screening technology and applied it to clinically relevant targets. We have also used this approach to enable the discovery of selective eukaryotic translation inhibitors—a potentially general strategy for probing the proteome. We are embedding beads in 3D tissue culture to induce and detect cell signaling changes, enabling the first phenotypic cell-based DEL screens and raising the possibility of screening in complex tissue niches, like tumor microenvironments or transport through the blood-brain barrier. We have also shown that we can leverage proximity to synthesize mRNA display OBOC beads, and we are interested in understanding if mRNA display beads can be similarly used in biochemical and cellular activity-based screens. Automation and accessibility are key themes. Finally, we are engineering magnetic bead-based methods to automate library synthesis, gel-phase biochemical assays, bulk emulsification-driven reaction compartmentalization, and FACS screening to dispense with complex microfluidics. This automated and accessible platform could broaden participation in drug discovery while unlocking the ability to probe the proteome.

NIH Research Projects · FY 2025 · 2021-04

RESEARCH SUMMARY Embryonic development involves the formation of functional organs comprised of many different cell types, which often originate from different locations. Thus cell migration and differentiation must be tightly coordinated, but are typically studied as independent processes. Here we use a combination of gene expression profiling in single cells, zebrafish genetics and computational models to test the hypothesis that migration and differentiation are coordinated. This coordination requires specific regulators of cell adhesion dynamics and cell-cell signaling. We focus on neural crest cells, a transient embryonic population that migrates throughout the body to give rise to a huge range of different fates. One barrier to studying this problem has been the limited number of tools available to detect transitional states in individual cells as they differentiate and tie this to their migratory behaviors in a precise and quantitative manner. Here we develop new approaches for profiling gene expression in single cells from known locations and tracking their movements in vivo. We will analyze how genes required for cell adhesion and cell-cell signaling influence these processes, and use computational models to predict key features of neural crest cell responses. We expect that such a multidisciplinary approach will reveal insights into the mechanisms that integrate cell migration and fate.

NIH Research Projects · FY 2025 · 2021-04

Summary/Abstract: University of California Irvine (UC Irvine) is driven to educate, train, and mentor academically oriented medical students who will pursue research and become future leaders in medicine. We believe that, “there is no science tomorrow without training today.” We believe that engaging medical students in research is of fundamental importance because: 1) it will enhance their ability to more critically evaluate the validity and efficacy of old and new emerging medical treatments; 2) ipso facto it will better equip them to fulfill their moral and ethical obligations to provide the best care for their patients; and 3) it will help uncover the “hidden gems” of medical students that will go on to pursue formal careers as physician-scientists. Therefore, in this NIDDK T35 application we propose to engage MD students in hands on research, mentoring, publishing, career development, and leadership in our Summer Mentored Research Training Program (SMART DDK). Through our expertise in a multidisciplinary “cells to society” approach, we aim to enhance the diversity and availability of physician scientists to meet the existent and evolving global challenges for improving health by the following specific aims: 1) providing outstanding leadership and a rigorous process of trainee-mentor alignment; 2) creating a flexible and innovative curriculum that emphasizes both core competencies and advanced concepts in NIDDK related research; and 3) maximizing access to the training program. These three specific aims will emphasize the importance of: 1) Claude Bernard’s “scientific medicine;” 2) Sir Issac Newtons sentiment of “standing on the shoulder of giants;” and 3) mentoring as reflected by perspective that the quality of science tomorrow is determined by the training quality of today. Within this context, our trainees will participate in a Trainee-Mentor Alignment Phase where they will identify a primary mentor, construct an individual development plan (IDP), and develop a research proposal. The oversight of trainee-mentor alignment, research proposal, and IDP will occur at regular intervals via formal presentations by the trainee to the Program Directors, Administrator, and mentor. Additionally, trainees will participate in formal courses, workshops, journal clubs, and grand rounds to develop a deeper appreciation for issues related to innovation, foundations of research, and cutting-edge technologies relevant to the realm of NIDDK related research. Issues related to the responsible conduct of research and reproducibility will be interwoven throughout all didactic components of the program. In addition to providing guidance to our trainees during their MS1 year, we will also continue to longitudinally follow their progress and engage the students in research during MS2- MS4 years via our medical student research program.

NIH Research Projects · FY 2025 · 2021-04

Project Summary/Abstract Neurodegenerative disease mechanisms encompassing Alzheimer’s disease (AD) are complex and have remained largely unknown to date despite the identification of risk factors and genetic mutations associated with the disease. Oligodendrocytes (ODCs) have critical roles in the central nervous system where they ensheath axons to allow rapid saltatory conduction and also provide metabolic support to neurons. Although a largely homogeneous oligodendrocyte population is thought to execute these functions throughout the brain, recent reports suggests that ODCs are highly diversified cell populations in the brain. Using systems biology approach, that integrates multi-omics data from human pathological specimens and mouse models of AD, this project aims to generate single-cell resolution genome-wide maps of transcriptional and regulatory networks in ODCs associated with AD. The project represents a major advance in the field by taking a comprehensive approach to data-driven discovery to identify highly conserved regulatory programs of AD. In parallel, the project aims to understand the role of sterol regulatory-element binding proteins (SREBPs) in AD using functional genomic approaches. Finally, the project will identify novel cell-type specific transcriptional regulators of AD using cutting edge approaches to data science.

NIH Research Projects · FY 2025 · 2021-03

PROJECT SUMMARY/ABSTRACT In the most severe cases of epilepsy, where seizures persist despite multiple trials of anti-seizure medications, patients may benefit from surgical removal of seizure-generating brain tissue. Prior to surgery, electrodes are often implanted directly into or onto the patient’s brain and are used to continuously record electrical brain activity over days. This is done to capture seizure activity and determine its point of origin, i.e. the seizure onset zone. If the seizure onset zone is identified, clinicians then use this information, in combination with the results of brain imaging and other testing, to guide removal of the corresponding brain tissue. While epilepsy surgery may lead to seizure freedom, 70-90% of surgery patients remain on anti-seizure medications and roughly 50% of patients continue to have seizures. The long-term goal of this work is to improve the outcomes of patients undergoing epilepsy surgery by developing more accurate methods to localize seizure-generating tissue. High frequency oscillations (HFOs) have garnered considerable excitement for their potential to identify and localize epileptogenic brain tissue. HFOs are short bursts of high-frequency electrical activity that occur in the brains of patients with epilepsy. They occur more frequently in the epileptogenic zone (EZ, the hypothetical area that must be excised to attain post-operative seizure freedom), and surgically removing HFO-generating brain tissue increases the likelihood of seizure freedom. While ongoing clinical trials are attempting to assess their prospective value for epilepsy surgery planning, there are multiple barriers to their widespread use. While group-level results are robust, HFO analysis is not yet predictive for single subjects. Recordings lack the sensitivity to reliably measure HFOs in every patient, and the occurrence of non-epileptic HFOs confounds the results. Therefore, HFOs are poised to revolutionize epilepsy surgery, but there is a critical need to optimize their measurement and maximize single-subject accuracy. The overall objective of this proposal is to improve EZ localization accuracy through systematic determination of the optimal HFO measurement methodology, coupled with novel, robust methods for HFO analysis. The rationale is that developing these novel methods with improved measurement techniques will increase the accuracy and robustness of HFOs as a biomarker of the seizure onset zone, thus improving the surgical management of epilepsy. To attain our objective, we will pursue three specific aims: (1) Demonstrate that electrode size is a crucial factor in HFO measurement. (2) Develop an automated method for patient-specific localization of the EZ based on HFOs. (3) Evaluate the effects of electrode size and HFO analysis method on EZ localization. The proposed research is significant because it will provide specific recommendations for the measurement and analysis of HFOs, enabling accurate, detailed localization of epileptogenic brain tissue. The expected outcome of this work is that it will guide surgeons in choosing which brain regions to remove and will increase the pool of potential surgical candidates. Overall, this will have a positive impact by leading to a greater chance of seizure freedom and improved quality of life for patients with the most severe cases of epilepsy.

NIH Research Projects · FY 2026 · 2021-03

PROJECT SUMMARY/ABSTRACT Long-term memory impairment significantly impacts patients with temporal lobe epilepsy (TLE), but no targeted treatment exists. There is a critical need to understand the mechanisms that disrupt long-term information stor- age in the epileptic brain. The long-term goal is to develop strategies to ameliorate and prevent cognitive impair- ment in patients with epilepsy. The overall objectives are to i) elucidate how interictal epileptic discharges (IEDs) affect hippocampal-cortical communication during memory consolidation, and ii) determine how closed-loop modulation of this communication alters long-term memory in an animal model of TLE. The central hypothesis is that IEDs disrupt the interaction of hippocampal ripples and cortical sleep spindles, altering neural activity patterns and plasticity. Further, restoration of physiologic hippocampal-cortical coupling can normalize the long- term memory deficits observed in TLE. The rationale for this project is that elucidating the spatiotemporally spe- cific network mechanisms that underlie long-term memory disruption will enable prevention of pathologic com- munication without impairment of physiologic communication, thereby facilitating memory consolidation. The central hypothesis will be tested by pursuing three specific aims in a TLE animal model: i) evaluate the effect of modulating hippocampal-cortical oscillatory coupling on neural spiking patterns; ii) determine the effect of mod- ulating hippocampal-cortical coupling on long-term memory, iii) establish links between large-scale molecular markers of synaptic plasticity and hippocampal-cortical oscillatory coupling during impaired memory consolida- tion. To accomplish these aims, in vivo electrophysiology and assays of immediate early gene expression will be paired with closed-loop electrical stimulation in freely behaving rats performing long-term memory tasks. The research proposed is innovative, in the applicant’s opinion, because it focuses on modulating neuronal commu- nication between brain regions during the interictal period to enhance memory consolidation. These contributions will be significant because they are expected to provide the mechanistic groundwork for development of novel approaches to treat, and potentially prevent, memory impairment in patients with epilepsy.

NIH Research Projects · FY 2025 · 2021-03

Project Summary / Abstract Blood vessels, from arteries to capillaries to venules and then to veins, contribute to fundamental physiological processes. However, the vascular responses for repair and restoration of microvascular networks after cortical brain injury are poorly understood, including how neuronal activity influences these processes. A major barrier to research is the poor accessibility of micro-vessels in the brain and associated technical difficulties. To overcome this barrier, we propose to use innovative imaging technologies that we have co-developed to investigate micro-vessel formation and re-growth in response to focal cortical injury. We have used light-weight head-mounted, miniaturized microscopes (“miniscopes”) to dynamically image the vasculature and associated cells with high spatial and temporal resolution. We will use cortical injury models by applying a controlled moderate impact to the mouse motor cortex. Combining in vivo longitudinal miniscope and 2-photon imaging, histological “vessel painting” and perfusion-weighted magnetic resonance imaging (PWI MRI), we aim to achieve a deeper understanding of microvascular restoration following cortical injury. We will apply targeted optogenetic stimulation of excitatory neurons and specific inhibitory neurons to modulate microvascular repair in early and late phases of vessel re-growth. Our guiding hypothesis is that microvascular restoration and remodeling after cortical injury are regulated by vascularization sequences and cellular processes that are similarly observed in normal vasculogenesis during central nervous system development. In Aim 1, we will identify the time course and spatial pattern of vascular regrowth, and blood flow dynamics after focal cortical injury. Vascular networks and blood flow are visualized with fluorescent-labeled dextrans for in vivo imaging for quantitative measurements. In Aim 2, we will determine the role of endothelial cells in new blood vessel sprouting and the establishment of functional microvascular by imaging Tie2-Cre reporter mice during the first two weeks post- injury. We will also examine the influence of astrocytes and pericytes in vascular re-growth. In Aim 3, we will test the hypothesis that optogenetic stimulation of specific neuron types in a temporally controlled manner facilitates and enhances microvasculature restoration for post-injury repair. We will also examine if and how targeted modulation of neural activities modulate Wnt/ß-catenin and VEGF signaling mechanisms that are critical for micro-vessel re-growth. Behavioral testing will assess the outcomes of the optogenetic treatment. We have strong preliminary data that supports the premise for the proposed research for all aims. The proposed research will advance our understanding of the cellular and molecular mechanisms underlying cortical microvascular restoration and how neural stimulation enhances vascular network formation.

NIH Research Projects · FY 2026 · 2021-03

ABSTRACT The major obstacle to successful cancer therapy is the rapid development of drug resistance. While targeted therapies often extend overall survival in the subset of patients with sensitizing mutations, their effects are short-lived. Patients who initially respond to these drugs generally develop resistance within a few months. Single-cell sequencing of tumors has revealed significant genetic heterogeneity; tumor cells without the sensitizing mutation survive therapy and re-populate the tumor. At the same time, compensatory epigenetic and genetic changes relieve dependence on the targeted pathway, also contributing to resistance. There is thus a critical unmet need for new therapeutic strategies capable of providing more robust cancer control. A robust system continues to function even when an individual component fails. In the context of drug development, a robust therapy would produce parallel, redundant anti-cancer effects, each of which is sufficient to inhibit tumor growth. One approach to achieving such redundancy is to embrace the pleiotropic actions of natural compounds. Endogenous signaling molecules produce coordinated and complex responses by targeting multiple signaling nodes in parallel. For example, endogenous sphingolipids exhibit potent tumor suppressor activity by producing multifaceted and incompletely characterized changes in signaling pathways that trigger proliferative arrest in normal cells and death in cancer cells. SH-BC-893 (893), a synthetic sphingolipid with improved drug properties, retains the anti-neoplastic activity of these natural compounds. In a rigorous, genetically-engineered mouse model for aggressive prostate cancer, 893 reduces autochthonous tumor growth by 82%. In a related subcutaneous isograft model, 893 produces tumor regressions in >50% of mice. 893 is also effective against patient-derived prostate tumor organoids that are resistant to standard-of- care therapies. The major argument against pleiotropic agents has been that toxicity will be unacceptably amplified relative to more specific drugs. However, natural sphingolipids induce quiescence in normal cells as part of an adaptive, homeostatic response to stress. Indeed, 893 does not cause organ toxicity or disrupt the proliferation of normal cells in the bone marrow or intestinal crypts even after 3 months of treatment with the anti-neoplastic dose. Normal cells are more resistant to 893, but 893’s pharmacokinetic properties also likely contribute to its safety margin. Our preliminary data showing that 893 engages multiple, high-value oncology targets results raise the possibility that 893 will be less susceptible to drug resistance and could overcome resistance to FDA-approved therapies. This proposal will test this provocative hypothesis. The expected results would have a significant positive impact by changing thinking in the field and providing a novel therapeutic strategy that would be effective in patients with late-stage, lethal prostate cancers.

NIH Research Projects · FY 2026 · 2021-03

PROJECT SUMMARY: Differentiation of progenitor cells requires their ability to interpret genomic information in diverse and highly specific ways. Understanding how gene batteries are regulated during this transition is one of the most fundamental questions in biology. A particularly intriguing context for this question is the process of zygotic genome activation (ZGA), when the paternal and maternal genomes unite after fertilization and begin transcription for the first time to transcribe its genome, this marking the initiation of the embryonic developmental program. During this stage, maternal factors such as transcription factors and chromatin modifiers that are present in the egg, are recruited to the embryonic genome to coordinate its first wave of gene expression and begin shaping its epigenetic and transcriptional landscape. Uncovering how the naïve embryonic genome is gradually remodeled into defined active and repressive domains is a central challenge in developmental biology. Recent studies have also revealed that some aspects of this regulatory process may be inherited from the parental genomes, suggesting that specific features of the maternal and paternal epigenomes contribute to early embryonic regulation. Our central hypothesis is that maternal components play a critical role not only in initiating embryonic transcription, but also in establishing the epigenetic states of the early genome through physical interactions with histone-modifying enzymes. However, dissecting these processes is challenging due to limited embryonic material for structural and genomic analyses and the technical difficulty of generating maternal mutants. Two recent advances now make this investigation feasible. First, we have developed an efficient CRISPR/Cas9-based strategy to generate biallelic maternal mutants. Second, we have established access to a highly sensitive cryoEM approach that dramatically improves structure analysis. Our project will illuminate the coordinated interplay between maternal transcription factors and epigenetic modifiers during the earliest stages of development. Leveraging the strengths of the Xenopus system including amenability to mutagenesis, biochemical accessibility, and compatibility with high-resolution cryo-EM, we will integrate molecular genetics, structural biology, and genomics to dissect maternal TF function, enhancer–promoter looping, and 3D chromatin architecture during ZGA. At the end of the fifth year we expect to provide major mechanistic insights into the process of ZGA.

NIH Research Projects · FY 2025 · 2021-02

Project Summary Clostridioides difficile (C. difficile) is a major opportunistic pathogen that colonizes the colon when normal gut microbiota is disrupted. The large protein toxin TcdB is a major virulence factor responsible for diseases associated with C. difficile infection (CDI). However, prior efforts to develop neutralizing monoclonal antibodies and vaccines against TcdB have yielded unexpectedly low efficacy or even failure. We believe that a key weakness of these previous studies might be the complexity of toxin variations seen clinically. While a single toxin sequence from a reference strain has been widely used in all previous therapeutic development, sequencing TcdB in clinical isolates in recent years has revealed a growing number of C. difficile strains as well as variations in toxin sequences. This may account for the reduced neutralization efficacy of the only FDA- approved monoclonal antibody, bezlotoxumab, against some TcdB variants such as the one produced by a hypervirulent strain (ribotype 027). The sequence variation and the toxin’s large size (~270 kDa) also pose daunting challenges to develop effective vaccines using the traditional toxoid approach. Building on our recent progress in identification of toxin receptors and understanding the structure and function of TcdB, here we propose to develop receptor-decoy-based therapeutic proteins as broad-spectrum antitoxins and a new generation of epitope-focused fragment-based vaccines, which could provide effective protection against most of the known TcdB variants. Frizzled proteins (FZDs) and CSPG4 are two major host receptors for TcdB, and we previously have revealed the mechanism by which TcdB recognizes FZDs. The first aim in this project is to establish a structural understanding of TcdB binding to CSPG4. Our second aim will focus on design and characterization of a family of bi-specific receptor-decoy proteins, which are composed of the optimized TcdB- binding fragments of CSPG4 and FZDs. In the third aim, we will take advantage of our knowledge of the structures of TcdB holotoxin, TcdB–antibody complexes, and TcdB–receptor complexes to design candidate vaccines based on the selected highly conserved and functionally critical TcdB fragments. This project is built on long-standing productive collaborations between the Jin lab and the Dong lab, combining their highly complementary expertise in structural biology and protein engineering (Jin lab) and TcdB receptors/CDI pathogenesis/animal models (Dong lab). Successful completion of this project will provide prototypes of antitoxins for immunoprophylactic therapy and broad-spectrum candidate vaccines that offer prophylactic and long-lasting protection.

NIH Research Projects · FY 2025 · 2021-02

Project Summary Cataract, the opacification of the eye lens, is the leading cause of blindness worldwide. Aquaporin 0 (AQP0), the most abundant membrane protein in the lens, functions as a water channel and as an adhesive protein. Defects in AQP0 can produce cataract, as well as have adverse effects on lens development. Despite its critical role in lens physiology, the functions of AQP0 are not fully understood. Our proposed research seeks to advance our understanding of how AQP0 water permeability (Pf), the exquisite control of which is required to maintain lens clarity, is regulated by Ca2+ and protons, whose concentrations depend on the AQP0 location within the lens. The proposed studies also seek to identify the amino acid residues that are crucial for Pf regulation and for the adhesive function of AQP0, through protein-protein and/or protein-membrane interactions, and to determine the effects of genetic modifications of AQP0 on lens physiology and development. To these ends, we will employ a tightly coupled, multi- disciplinary approach, unique within the field of aquaporin research, which employs techniques ranging in scale from the atomic/molecular to the cellular and organismal level. Specifically, in Aim 1 we propose to use in vitro Xenopus oocyte permeability measurements on a panel of mammalian and fish AQP0 mutants to assess the contribution of particular residues to AQP0 Pf and its regulation, along with adhesion assays on lens fiber cells from zebrafish containing wild-type and mutant Aqp0s. These experimental approaches will be complemented in Aim 1 with in silico multi-µs molecular dynamics simulations, validated by comparison with experimental Pf measurements, to elucidate mechanistic aspects of the influence of Ca2+ via calmodulin binding, pH, including the effects of a variety of strategically chosen mutations on the Pf of mammalian and fish AQP0s. The computer simulations proposed in Aim 1 will also address the relative role of protein-protein and protein-membrane interactions in the adhesive function of AQP0. Aim 2 will use genetically modified zebrafish to determine how AQP0 contributes to the structure and function of the lens and its development in vivo. Our work will improve understanding of lens physiology and the molecular mechanisms of water channel gating and its regulation by Ca2+ and pH, and uncover fundamental principles that could inform the future development of therapeutic strategies for delaying or eliminating cataract formation.

NIH Research Projects · FY 2025 · 2021-01

Age-related decline in central auditory function significantly affects quality of life in the elderly, with impaired speech perception leading to increased risk for depression, social isolation and cognitive decline. A 2017 Lancet Commission report cites hearing loss as the largest modifiable risk factor for developing cognitive decline, exceeding smoking, high blood pressure, lack of exercise and social isolation. Remarkably, a 2019 large-scale study found that even mild hearing loss, i.e., still within the normal range, produced an even closer association with cognitive decline. Currently, there is no effective therapy for age-related central auditory decline—hearing aids only address audibility—and no drug treatment. Ideally, a combination of drug treatment with hearing aids and behavioral training could restore auditory function, but the development of pharmacological treatments requires a better understanding of the mechanisms by which candidate drugs improve hearing. The goals of this proposal are to develop biomarkers of altered auditory processing in aging mice and humans, and using these biomarkers, to test the hypothesis that nicotine can normalize these age-related central auditory deficits. Because nicotine enhances cortical and cognitive function, pharmaceutical companies are developing nicotine-like drugs to target cognitive deficits in aging. These drugs are non-addictive (unlike nicotine in tobacco), yet nicotine also is non-addictive when given topically or orally. However, its clinical benefits have not been exploited except as an aid to stop smoking. We hypothesize that: 1) acute nicotine compensates for the age-related decline in inhibition by exciting the remaining inhibitory neurons; 2) chronic nicotine exposure (CNE) upregulates nicotinic acetylcholine receptors (nAChRs); and, as a result, 3) acute nicotine and/or CNE will reduce or reverse the age-related auditory decline. We will test these hypotheses in both mouse and human at the level of cells (mouse in vitro brain slice), neural systems (mouse in vivo physiology; human brain imaging and EEG) and behavior (human psychoacoustics). Aim 1 will determine in mouse whether age-related decline in auditory spectrotemporal processing is reversed by acute nicotine or CNE, and characterize the associated cellular mechanisms. Aim 2 will identify, in humans, age-related changes in receptive field properties in auditory cortex using novel fMRI techniques and determine if nicotine reverses these changes using psychoacoustics, fMRI and EEG. This project features a multifaceted, parallel approach in mouse and human. Each Aim will: 1) examine auditory processing at multiple adult ages; 2) use similar acoustic stimuli in both species, accounting for species differences in hearing, to target common mechanisms; 3) test the effects of nicotine. A successful outcome will promote an integrated understanding across levels, from cellular mechanisms to perception, and facilitate translation of nicotine-based therapeutic treatments to clinical populations.

NIH Research Projects · FY 2024 · 2020-12

Every 4.5 minutes, a baby is born with a birth defect in the United States. Skeletal defects of the bony skeleton have been associated with environmental chemical exposure in utero. Our lab has shown that proper development of osteoblasts, the bone forming cells, depends on tight regulation of bone-specific genes. We developed a human embryonic stem cell (hESC) system where toxicant-induced differential gene expression perturbed osteoblast differentiation. MicroRNAs (miRNAs) are small non-coding RNAs that epigenetically regulate gene expression and are actors of skeletal development. There is no knowledge whether miRNAs adversely respond to environmental agents to result in skeletal malformations. In this proposal, I hypothesize that toxicant-induced miRNA changes play a critical role in the manifestation of skeletal birth defects. MiRNA profiling on differentiating hESCs previously identified 10 miRNAs downregulated stemming from chemical exposure that repressed osteoblast differentiation. I propose to 1) validate the functional role of candidate miRNAs during osteogenesis (K99); 2) investigate whether the functional effects of the candidate miRNAs are replicated in vivo (K99); 3) determine the mRNA target genes and signaling pathways affected by the candidate miRNAs (K99); and 4) investigate whether miRNA dysregulation is facilitated through epigenetic machinery that represses miRNA expression (R00). The proposed aims will significantly impact the understanding of environmentally induced epigenetic toxicity during skeletal development. Mentors Drs. Martin Riccomagno (survival surgeries and genome engineering) and Nicole zur Nieden (hESCs, miRNAs and genome engineering) will aid in my training for successful completion of the proposed aims and becoming an independent principal investigator. Additional support will come from Drs. Martin Garcia-Castro (molecular cytogenetic techniques) and Patrick Allard (epigenetic toxicology). The career development plan includes training to enhance my research skills, including Next Generation Sequencing, bioinformatics, in utero electroporation, and in vivo skeletal analysis, as well as scholarly skills and professional development, including but not limited to science communication, writing, mentoring, and management skills. The planned training and research will facilitate the transition to independency and success as a principal investigator.

NIH Research Projects · FY 2026 · 2020-09

ABSTRACT High-acuity color vision relies on the fovea and macula in the central retina and is used for everyday tasks like reading and driving. Age-related macular degeneration (AMD) progressively destroys photoreceptors and retinal pigment epithelium (RPE) in the central retina, leading to severe vision loss. Evaluating new AMD treatments requires measuring their effectiveness and thus requires a profound understanding of functional changes in AMD. However, although AMD primarily affects the central retina, most preclinical studies rely on animal models that lack a macula and fovea, leaving the physiology of photoreceptors and RPE largely unexplored. Building on previous pioneering insights into rod and cone photoreceptor physiology in the human macula and peripheral retina, this project hypothesizes that central photoreceptors are specifically adapted to function under bright light conditions and that AMD disrupts some of these critical pathways of high-acuity vision. Aim 1 will measure phototransduction parameters, such as amplification constants and response kinetics, as well as light adaptation from human foveal, para-/perifoveal and peripheral rod and cone photoreceptors using suction electrode recordings. This aim hypothesizes that central photoreceptors show lower gain of the phototransduction activation reactions and weaker modulation of their light sensitivity by background light than peripheral photoreceptors. The impact of background light on rate- and non-rate-limiting phototransduction deactivations reactions will be tested. Aim 2 will examine RPE-dependent and -independent pathways contributing to dark adaptation in healthy and AMD-affected human maculae using ex vivo optical coherence tomography and fundus imaging, ex vivo electroretinogram, and retinoid analysis by high-performance liquid chromatography. This aim hypothesizes that that both RPE-dependent and -independent regeneration contribute to cone dark adaptation and pigment regeneration. Additionally, this aim hypothesizes that dark adaptation is delayed in cones from AMD patients. Through improved understanding of central and peripheral photoreceptor function under varying light conditions and in the pathology of AMD, this study will be critical for developing diagnostic tools, establishing functionally relevant clinical endpoints for clinical studies, and elucidating AMD disease mechanisms for therapeutic development. Ultimately, this will improve available AMD treatments and provide hope to millions of patients at risk of vision loss and associated diminished quality of life.

NIH Research Projects · FY 2024 · 2020-09

Project Summary / Abstract Age-related cognitive decline is an important concern in the United States, as approximately 20% of the US population is expected to be age 65 or older by year 2030. Understanding the molecular mechansims of brain aging to prolong healthy cognitive function is therefore increasingly important as the population ages and older people remain in the work force. Brain cells exhibit profound and heterogeneous changes during aging at molecular and cellular levels. The simple intervention of physical exercise has emerged as a major positive modulator of cognitive function in aging. In response to RFA-RM-20-005, we have formed an interdisciplinary team with expertise in single-cell genomics, neural circuitry, and aging, to investigate age- and physical activity- related changes of 4D nucleome in post-mortem human brain hippocampus cells across the lifespan with single- cell resolution. We hypothesize that cell-type-specific re-organization of nucleome occurs in the human hippocampal brain region during aging and with physical activity. The changes in nucleome in turn control brain epigenome and transcriptome, modulating neural circuit functionality. The “Methyl-HiC”, a new approach for joint profiling of DNA methylation and chromatin contacts in single cells, combined with “Paired-seq”, an ultra- high-throughput method for single-cell joint analysis of open chromatin and transcriptome, will be used to interrogate the chromatin architecture along with DNA methylation, chromatin accessibility and gene expression in the human hippocampus. In Aim 1, we will determine changes in nucleome in major cell types of post-mortem human hippocampus across the life-span with 4 age ranges (20–39, 40–59, 60–79, and 80–99 years old). We will further correlate these changes in nucleome with epigenome and transcriptome in each cell type, to identify vulnerable cell types during aging, and uncover potential gene regulatory programs that could be impacted by aging. In Aim 2, we will determine how physical activity modifies and restores nucleome in specific human hippocampal cell types. We will study two age-matched cognitively–healthy cohorts (70-99 years old) with either high level or low level physical activity, as measured by wearable activity monitors. We will correlate restorative effects on nucleome with epigenome and transcriptome. In Aim 3, we will map how aging and exercise alter nucleome in specific hippocampal cell types with highly controlled quantifiable physical activity in the mouse model, for comparison with human data. These mouse studies allow the exercise variable to be investigated in isolation from effects of other lifestyle factors that can affect hippocampal nucleome, which is not possible with human subjects. The proposed research will help to transform our ability to understand the mechanisms of chromatin organization and function in the context of human brain aging.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY The long-term goal of our research is to determine how the immune system interfaces with cells in the dystrophic niche to impact the pathogenesis of Duchenne muscular dystrophy (DMD), to provide a foundation for developing novel treatments for this devastating disease. The objective of the current study is to define how regulatory T cells (Treg) regulate macrophage-stromal interactions in muscle, and how these interactions can be pharmacologically manipulated to alter disease progression. We provide strong scientific premise for proposing the central hypothesis that Tregs regulate the severity of DMD by suppressing a distinct galectin-3+ muscle macrophage population that promotes fibrosis. This hypothesis is supported by our preliminary data and prior research, showing that 1) dystrophic muscle is populated by a putative fibrogenic galectin-3+/Spp1+ macrophage that we identified by single-cell RNA sequencing (scRNAseq); 2) prior research showing that spp1 promotes fibrosis in mdx mice; 3) macrophage-specific deletion of Spp1 in mdx mice causes remarkable changes in the transcriptional profile of PGFRa+ stromal cells, as assessed by scRNAseq; 4) galectin-3+ macrophages are expanded when Tregs are depleted in mdx mice, suggesting that Tregs suppress fibrogenic macrophages and 5) depletion of Tregs in mdx mice causes an increased expression of pro-fibrotic genes. We will test our central hypothesis by addressing two specific aims. Specific aim 1 will define how galectin-3+ muscle macrophages promote fibrosis. Two subaims will be addressed under specific aim 2, identify the molecular basis for Treg suppression of galectin-3+ muscle macrophages. In subaim 2a we will determine whether the Treg-mediated suppression of IFNg-producing cells is IL-10 dependent; and in subaim 2b we will examine whether the therapeutic expansion of Tregs suppresses fibrosis. Although a role for Spp1 in promoting fibrosis in mdx mice has been documented, a mechanism for this pathological process is largely unknown. Thus, our proposed study is significant as it will define the cellular basis by which Spp1 promotes muscle fibrosis, and advance our understanding of how Tregs ameliorate the severity of muscular dystrophy. The proposed research is innovative because it will define mechanisms of cellular cross talk that promote fibrosis in dystrophic muscles. The proposed studies use innovative technologies (e.g. scRNAseq) and experimental therapeutics (e.g. IL-2c) that will provide the framework for developing novel approaches to inhibit the development of fibrosis. These potential discoveries will advance the field by establishing interactions between the lymphoid and myeloid compartment of the immune system that operate in DMD to suppress or limit fibrogenic responses. Further, findings from the proposed studies are expected to have significant clinical implications for DMD, as the immunosuppressive function of Tregs can be therapeutically augmented to ameliorate disease severity by inhibiting pro-fibrotic inflammatory responses.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY Macrophages are central regulators of inflammation and tissue healing following injury or infection, and during disease. While much is known about how soluble, biochemical factors in the environment regulate macrophage function, less is known about how biophysical cues regulate their response, despite the fact that these cells exist within solid tissues that are rich in mechanical cues. Furthermore, many diseases in which macrophages are involved, such as cancer and fibrosis, are characterized by changes in tissue biophysical properties. Our previous work demonstrated that adhesion to soft extracellular matrix hydrogels inhibits macrophage inflammatory activation. In preliminary work, we found that matrix rigidity influences the localization of YAP, a transcriptional co-factor involved in cell proliferation, organ size control, and cancer, but with previously undescribed role in macrophage activation. Adhesion to stiff substrates leads to YAP nuclear localization, which appears to prime macrophages for a potent inflammatory response. In addition, cytoskeletal polymerization and the mechanically-activated and calcium-permeable ion channel Piezo1 appear to be involved in YAP nuclear localization and inflammatory activation. In this study, we propose to investigate the molecular mechanisms underlying YAP signaling and Piezo1 activity in the macrophage response within different stiffness environments. In Aim 1, we will examine the effect of stiffness on cytoskeletal remodeling and associated signaling pathways on YAP activity. In Aim 2, we will probe the role of Piezo1-mediated calcium activity in stiffness sensing, YAP signaling, and macrophage function. Finally, in Aim 3, we will investigate the role of YAP and Piezo1 on macrophage-mediated wound healing in vivo using a murine subcutaneous biomaterial implant model. An improved fundamental understanding of how macrophages sense their mechanical environment may lead to new immunomodulatory strategies that control macrophage function during disease.

NIH Research Projects · FY 2024 · 2020-09

Project Summary Lyme disease, one of the most commonly reported infectious diseases in North America, is caused by the tick-borne bacterium Borreliella burgdorferi. Although humans and other large mammals can be infected by B. burgdorferi, in order to complete its life-cycle in the wild the bacteria relies on rodent reservoirs, the major one being Peromyscus leucopus, the white-footed deermouse. The role of P. leucopus in Lyme disease and several other tick-borne diseases is analogous to that of bats as reservoirs for SARS coronaviruses and Ebola virus. In this proposal we continue the development of P. leucopus as an emerging genetic model system for the study of infectious and other diseases by maintaining and expanding genomic and biological resources for this species. These resources are the starting point for any gene-focused experiments in the Peromyscus genus. The primary goal of this proposal is to identify segregating genetic factors that impact the competence of P. leucopus as a reservoir of B. burgdorferi. The trait of reservoir competence is measured as the prevalence of infection and corresponding bacterial burdens among a cohort of nymphs that had molted from larvae previously fed on experimentally-infected deermice. Secondary endpoints include rates of growth and decline of the bacteria in the blood and skin of the animals and selected host responses, such as antibodies to the agent and inflammation of tissues, over the time course of the infection. It would normally be extremely difficult to carry-out large-scale genotyping and/or genetic crosses in an emerging rodent model. Here we show that our genome assembly for P. leucopus in concert with low pass short read sequences from a long-term closed colony of deermice can be leveraged to accurately impute SNP and haplotype genotypes on a genome- wide scale. These genotypes are then used to identify genes contributing to the remarkable capacity of P. leucopus to serve as a key reservoir host for B. burgdorferi and other disease agents. Finally, a subset of identified genes will be validated via CRISPR/Cas9 gene knock-outs in P. leucopus spearheaded by the person who pioneered transgenics for this genus. The identification of reservoir competence mediating genes may suggest better interventions to block transmission and provide insights into the management of human infections.

NIH Research Projects · FY 2025 · 2020-09

PROJECT SUMMARY Obesity is a serious public health concern, largely because obesity and related disorders (e.g., cardiovascular disease, type II diabetes, hypertension, cancers, etc.) add more than $200 billion annually to US health care costs. The current clinical paradigm for obesity is one of energy intake versus energy expenditure, with clinical management focused on diet and exercise. Diet and exercise are important factors in obesity, particularly the energy dense Western dietary pattern, but they do not fully account for the obesity epidemic. US adults were 2.3 kg/m2 higher in BMI in 2006 than in 1988, even at comparable caloric intake and energy expenditure. Emerging evidence supports an important role for exposure to endocrine disrupting chemicals (EDCs)in obesity. We identified tributyltin (TBT) as an environmental “obesogen” - a chemical that leads to weight gain, in vivo. In utero exposure to environmentally-relevant levels of TBT increased fat depot weight, reprogrammed mesenchymal stem cells to favor the adipogenic fate and caused non alcoholic fatty liver disease in F1-F4 male offspring. We reproduced these transgenerational phenotypes in two independent experiments and found that male F4 descendents of F0 TBT-treated animals became obese when dietary fat was increased. This fat persisted after the animals were returned to normal low fat chow. TBT-treated animals and their descendents were resistant to fasting-induced fat loss, indicating that these animals do not mobilize fat to the same extent as controls during fasting. We found that fat in F4 male descendants of TBT treated dams showed persistent DNA hypomethylation in regions encompassing important metabolic genes such as the Lep gene, increased leptin mRNA expression, elevated plasma leptin levels, and that these hypomethylated regions in fat were less accessible in sperm chromatin of F3/F4 males. We proposed that these animals exhibited a transgenerational "thrifty phenotype" caused by altered chromatin structure and accessibility. We hypothesize that TBT exposure modifies the epigenome across multiple generations, sensitizing animals to weight gain and that this “thrifty phenotype” is revealed or exacerbated by increased dietary fat. Two specific aims are proposed: 1) How does TBT exposure exacerbate the effects of “Total Western Diet” leading to weight gain?, and 2) How does TBT exposure make animals resistant to fat loss? Answering these key questions will address knowledge gaps in the field that are relevant to human health. The proposed research will reveal which molecular mechanisms may underlie the effects of obesogens and how a Western dietary pattern interacts with obesogen exposure to predispose toward fat gain and promote the transgenerational programming of obesity. This will greatly inform the thinking of clinicians and the public in understanding individual susceptibility to obesity and how best it may be treated and prevented in individuals. The successful completion of this research will illuminate the molecular mechanisms underlying the role of xenobiotic chemicals on obesity, and may provide insights into how the obesity epidemic can be curtailed.

NIH Research Projects · FY 2024 · 2020-09

ABSTRACT Over 5.8 million Americans are living with Alzheimer's dementia, a disease with no effective treatment and no cure. Two-thirds of the caregivers for persons with dementia (PWD) are women (most often family) and a third are themselves over 65. Dementia takes a significant toll on caregivers, often resulting in chronic stress, depression, sleep disorders, poor health related quality of life (HRQOL), and early mortality due 24/7 care responsibility for PWD. Research has shown significant barriers to dementia care for underserved populations, including Latinos and Asian minorities. Underserved family caregivers for PWD tend to underutilize public health services available, and do not seek treatment until the situation is unmanageable with current resources reporting barriers that included language, time, and finances. Monitoring the caregiver's health and wellbeing is important as well as their maintaining a positive interaction with the PWD. Thus, there is a need for an innovative and feasible intervention to improve underserved caregiver's mental and physical health. Little research is reported for dementia caregiver interventions in underserved minorities and one given at home by community health workers (CHWs). The proposed intervention meets the needs of these family caregivers in developing a positive relationship with the PWD by educating caregivers to better understand the PWD's behaviors. Another component of the intervention is stress reduction techniques, including mindful deep breathing and compassionate support/listening to reduce depression and improve family relationships making the caregiving less burdensome. By monitoring the physiological responses of stress (i.e. heart rate variability), sleep and activity, we can objectively measure changes as a result of the intervention. Using Wearable Internet of Things (WIoT) technology, a combination of Watch/ring-Smartphone-Cloud, has proven to be a significant method of monitoring behavioral and physiological measures providing evidence of change over time and uniquely associated with this intervention. Our preliminary data show that the intervention with WIoT brought to the caregiver by CHW home visitors was acceptable to ethnic caregivers (Latino, Vietnamese, and Korean) and effective in reducing caregiver stress and burden over the short term. With the addition of non-Hispanic Whites, the proposed caregiver-centered, culturally and language appropriate, CHW home-visit-based 3-month intervention has 3 significant parts:1) stress reduction by mindful breathing and compassionate support/ listening to improve caregiver's health and well-being; 2) education on caregiving skills to improve responses to the PWD and in turn their behaviors; 3) WIoT physiological and behavioral monitoring. This 6-month randomized controlled trial will compare outcomes (burden, depression, HRQOL, PWD behaviors, caregiving self-efficacy) between the intervention, attention control with use of the WIoT only, and usual care groups at baseline, 3 months, and 6 months. This intervention using the CHW-model and WIOT technology has the potential to lessen health disparities in dementia caregiving in ethnic and underserved family caregivers.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY Current therapies do not alter the course of Alzheimer's disease (AD), a devastating neurodegenerative illness that affects 5.8 million people in the United States and 44 million people worldwide. Thus, an urgent goal for research is to identify points of control for neurodegeneration, which may be targeted to slow down AD progression. In this application, we propose to test the hypothesis that the enzyme N-Acylethanolamine Acid Amidase (NAAA) is one such focal point. NAAA is a lysosomal lipid hydrolase that converts palmitoylethanolamide (PEA) into palmitate. PEA is an endogenous agonist of the neuroprotective nuclear receptor PPAR-α, whereas palmitate promotes neurodegeneration by suppressing the transcription co- activator PGC1α, a key regulator of neuronal energy metabolism and survival. Preliminary experiments have shown that NAAA transcription is abnormally elevated in persons with sporadic AD and in various animal models of neurodegeneration, including the 5xfAD model of AD. In the same models, we found that pharmacological NAAA inhibition and/or genetic NAAA deletion exert marked protective effects. Based on these results, we hypothesize that dysfunctions in NAAA-regulated lipid signaling may be critically involved in the pathogenesis of AD. We have three specific aims. Aim 1. Characterize NAAA-regulated lipid signaling in mouse models of AD. Using 5xfAD and Tau P301S mice, two mouse lines that capture distinct aspects of AD pathology, we will identify age-dependent, regionally selective changes in NAAA- regulated lipid signaling, which might precede and/or accompany neurodegenerative alterations and cognitive impairment. Aim 2. Determine the impact of pharmacological NAAA inhibition in mouse models of AD. We will assess the impact of chronic administration of the compounds ARN19702 and ARN16186 – two brain-permeant NAAA inhibitors discovered by our team – on molecular, morphological and behavioral markers of disease progression in 5xfAD and Tau P301S mice. Aim 3. Determine the impact of genetic NAAA deletion in mouse models of AD. Using our conditional NAAA-/- mice, we will generate NAAA- deficient 5xfAD and Tau P301S mice to evaluate the impact of NAAA deletion on disease progression. In Aims 2 and 3, we will also explore molecular and cellular substrates for the effects of NAAA inhibition/deletion using single-cell RNA sequencing. The proposed studies will elucidate the functional roles of NAAA-regulated lipid signaling in the pathogenesis of AD and, if our hypothesis is verified, will validate NAAA as a novel molecular target for the treatment of this disorder.