University Of California-Irvine

NIH Research Projects · FY 2025 · 2022-09

Project Abstract Alzheimer’s disease (AD) is currently an incurable neurodegenerative disease that affects over 35 million people worldwide, including 5.4 million individuals in the USA with a new case diagnosed every minute. Amyloids occur commonly as key pathological features in a wide variety of neurodegenerative diseases such as AD and despite 30 years of intensive research, important questions about the significance, mechanisms and role in pathogenesis of amyloids remain unresolved. Because of the strong implication of amyloid Aß peptide in familial AD, preparations of amyloid aggregates (oligomers, fibrils) have been featured in over 100,000 publications over the past 30 years. In many cases, conflicting, contradictory and non-reproducible data obfuscate the underlying mechanisms of pathogenesis. Advances in structural biology have established the structures of several distinct types of amyloid, leading to the discovery that amyloid structures are highly polymorphic with the same protein sequence adopting distinct beta sheet folds. Moreover, different structures seem to correlate with different disease subtypes, indicating that structural variation plays a role in pathogenesis. This polymorphism and structural heterogeneity may also underly the irreproducibility and conflicting results that have plagued amyloid research. This suggests a critical need for well-characterized, standardized protocols for the preparation of different types of amyloid Aß aggregates and reagents to authenticate these preparations and specifically identify and quantify these polymorphs in vitro and in brain in order to support basic research to understand the roles and mechanisms of amyloids in disease pathogenesis. Therefore, the overall goal of this resource initiative is to establish a source network for the development of reagents and protocols for the production, standardization, characterization, authentication of amyloid oligomers and fibrils and the dissemination of these materials to investigators.

NIH Research Projects · FY 2025 · 2022-09

Supplementary Funding Request for U01 EY034594-01: "Correlating Genomic AMD Risk Variants with Lipid Composition and Phagocytic Function of Patient-Derived Induced Pluripotent Stem Cell (iPSC)- derived Retinal Pigment Epithelium (RPE)" The supplement instructions for PA-20-272 states the following: “At a minimum, the Research Strategy section should be completed and must include a summary or abstract of the funded parent award or project. Other sections should also be included if they are being changed by the proposed supplement activities.” The original Project Summary / Abstract document has not changed as a result of the proposed supplement activities.

NIH Research Projects · FY 2025 · 2022-09

Modified Abstract Section ABSTRACT Contextual factors related to social and economic conditions are the most salient determinant of population health differences. Although substantial progress had been made in understanding how contextual factors becomes biologically embedded, crucial knowledge gaps remain. Biological embedding has been described primarily at the lev-el of differentiated cell types and tissues. Based on the consideration that a) the long-term effects of contextual factors extend well beyond the lifespan of differentiated cells, whose replenishment occurs only from stem/progenitor cells, and b) can be perpetuated across generations, we advance the novel hypothesis that the origins of health differences may extend all the way down to the level of stem cells, and specifically to the effects of maternal exposure to contextual factors on offspring stem cells during prenatal life. Our proposed study will focus on differences between Hispanic, non-Hispanic Black, and non-Hispanic White mothers and their newborns in obesity and metabolic phenotypes; on mesenchymal pro-genitor/stromal cells (MSCs) and MSC-derived adipocytes as the stem and differentiated cells of interest; on mitochondrial function, adipogenic propensity/activity and insulin sensitivity as the key intracellular processes of importance; on newborn ad-ipose tissue mass and glucose-insulin regulation as the outcomes of significance; and on maternal-fetal gestational biology as the proximate transmission pathway. We will conduct this study in a cohort of N=240 child-mother dyads; isolate and culture fetal MSCs from newborn cord tissue; perform high-resolution cellular experiments; and characterize neona-tal phenotypes in vitro in MSC-derived adipoctyes, and in vivo using whole body densitometry. Aim 1 will test the hy-pothesis that maternal exposure to unfavorable contextual factors is associated with newborn mesenchymal stem cell characteristics, i.e., reduced mitochondrial efficiency, increased adipogenic propensity, and reduced insulin sensitivity. Aim 2 will test the hypothesis that variation in maternal and fetal gestational biology (composite measures of endo-crine, immune/inflammatory, and metabolic ligands) mediates the effects of contextual factors on newborn mesen-chymal stem cells. Aim 3 will establish the clinical significance of variation in MSC characteristics for neonatal obesi-ty-related phenotypes at the a) cellular level (MSC-derived adipocyte size and fat content; mitochondrial function; ad-ipogenic activity), and b) whole-body level (percent fat mass and systemic insulin sensitivity). Aim 4 (exploratory) will elucidate potentially modifiable maternal psychosocial and behavioral factors that relate to the specific components of contextual factors that are associated with newborn MSC biology. Aim 5 will establish a shared repository (bi-obank) of MSC, cord blood, cord and placental tissue samples for future studies of molecular mechanisms (gene ex-pression profiles, epigenetic characteristics) and in vitro cell differentiation analyses. Significance and impact: 1) Our study will define novel measures (with norms) in human newborn stem cells that profile the earliest vulnerabili-ties for health and population health differences; 2) broaden understanding of novel cellular targets and molecular mechanisms underlying biological embedding of contextual factors to inform risk identification, prevention, early di-agnosis, and personalized intervention; and 3) provide a unique and valuable shared resource (human newborn stem cell culture biobank).

NIH Research Projects · FY 2024 · 2022-09

Revised Project Summary Age and chronic inflammatory stress drive the emergence of mutant bone marrow stem cells that can lead to specific hematologic malignancy associated mutations allow HSC to resist specific stressors could be leveraged toward therapies aimed at neutralizing the selective advantage of the malignant clone. We have recently found that patients with myeloproliferative neoplasm (MPN) have a dampened response to the anti-inflammatory cytokine IL-10 and this leads to persistent production of the inflammatory cytokine TNFα. In a murine Jak2V617F MPN model, pharmacologic blockade of IL-10R signaling enhances the selective advantage of Jak2V617F mutant cells. The objective of this project is to define how JAK2V617F mutant HSC gain a selective advantage over wild-type (WT) HSC when IL-10R is blocked. Our central hypothesis is that dampened IL-10R signaling creates an inflammatory state that negatively affects WT but not JAK2V617F mutant HSC, thus endowing JAK2V617F mutant cells a selective advantage. Moreover, we hypothesize that dampened IL-10R signaling is an intrinsic feature of those predisposed to acquire MPN. In Aim 1 we will determine the prevalence of an IL-10 resistance phenotype among MPN families. We have found that MPN patients display an inability to respond to IL-10, which we term the “IL-10 resistance phenotype”. However, cases with a family history of MPN have not been systematically evaluated for the phenotype, nor have unaffected family members. We will determine the prevalence of the IL-10 resistance phenotype in affected and unaffected members of our growing MPN family registry which currently contains 53 MPN families. In Aim 2 we will identify the genetic basis for blunted IL-10R signaling in MPN. We will perform targeted DNA sequencing and gene expression profiling (i.e., RNA-seq, ChIP-seq, and differential DNA methylation analysis) to identify the underlying mechanism responsible for dampened IL-10 signaling in MPN. Using sorted peripheral blood monocytes we will use Illumina Infinium MethylationEPIC chips to determine if MPN patients and unaffected family members with blunted IL-10 signaling have differential methylation of IL-10R signaling genes. Gene expression profiling will be performed in parallel to determine if the methylation pattern accurately predicts gene expression. We will also perform targeted DNA sequencing, focusing on cytokine receptor pathway genes, and paying close attention to any genes found to have differential methylation and/or expression, to identify the genetic basis for blunted IL-10 signaling in MPN patients.

NIH Research Projects · FY 2025 · 2022-09

Abstract One of the most significant problems in the field of HIV deals with addressing the low rates of HIV care among individuals living with HIV/AIDS, especially among Black/African American and Latinx communities. This application seeks to study a novel way to address that problem by adopting and applying a cutting-edge “big data outreach” approach being used to increase consumer engagement by top technologies companies. This approach has recently been replacing other digital outreach methods largely for privacy reasons-- to conform to stringent European Union privacy laws-- as it involves de-identified data. The digital outreach method being proposed is already being applied in health (but not yet HIV) settings. During the COVID-19 pandemic, our team and others (including the CDC) studied and found success applying these methods for targeted digital recruitment and outreach to those at high-risk for COVID-19. As a result of the COVID-19 pandemic and its effect on use of digital/remote tools, these approaches will soon be applied to HIV to assist in targeting and engaging hard-to reach individuals in HIV research and care. Importantly, the proposed methods allow access to large-scale passively-collected, opt-in, community and mobility (GPS pings) data, which have been shown to add rich and granular data to improve health surveillance and interventions. This application seeks to use these novel digital outreach methods to identify and enroll individuals living with HIV/AIDS from communities of color who are at high-risk for being out of care, and analyze their mobility and community data to identify the key geographic contexts that impact HIV care engagement. We are conducting this effort for, and in partnership with, 2 Ending the HIV Epidemic (EHE) jurisdictions (Washington D.C. and Orange County health departments) and key participant stakeholders to gain their insights on needs, implementation (including ethical concerns), and potential future scale-up of this approach to improve surveillance and intervention efforts. Specifically, we seek to 1) Identify individuals of color living with HIV/AIDS within EHE regions who are at high-risk for being out of care, 2) Using GPS mobility, community (e.g., local crime), and HIV care data, identify the key geographic contexts that impact HIV care engagement, and 3) In partnership with the Washington D.C. and Orange County health departments, explore a case study of the ongoing barriers and facilitators of this approach at the individual, interpersonal, and structural levels. This 1-year cohort study will be focused on identifying people living within an EHE region who have been hard-to-reach for HIV care in order to converge with EHE outcome measures. To our knowledge, this is the first study to apply these novel “big data outreach” methods to HIV, will enroll the largest cohort to date with GPS mobility, community, and other HIV care contextual data, and the first HIV study to passively collect mobility data, which helps to increase data quality and reduce dropout rates compared to previous studies.

NIH Research Projects · FY 2025 · 2022-09

Abstract Multiplexed spatial profiling of protein markers in cells and tissues is critical to basic research and clinical applications. Unfortunately, we currently lack tools that can rapidly and routinely profile a large number of proteins in situ in large tissues with subcellular resolution in a time and cost-effective fashion. Existing tools for in situ protein analysis including immunohistochemistry and immunofluorescence suffer from low multiplexing because of limited separation of spectral channels. Recent single-cell sequencing methods lack the critical spatial context needed to understand complex heterogeneous samples. Other spatial proteomics methods that are based on serial labeling and imaging, indirect indexed mass spectrometry or sequencing are complicated, time-consuming and expensive. This proposed project will develop a new spatial proteomics technology termed as Phasor S- FLIM that enables direct, simultaneous, high-plex spatial profiling of protein markers in large and thick tissues with just one-round of staining and imaging. Our Phasor S-FLIM system, for the first time, allows true parallel, simultaneous lifetime and spectral detection with phasor analysis to obtain fast, unbiased, high-precision lifetime and spectral data that can be processed in real time. In the proposed work, we will adapt and further develop Phasor S-FLIM for high-plex spatial proteomics applications, including (a) implementation of Pulsed Interleaved Excitation (PIE) dual excitation with 2-photon lasers and sensitive multi-channel GaAsP PMT arrays, enabling exciting and detecting a broad range of fluorophores (Aim 1), (b) development of a novel fluorophore-quencher labeling strategy to generate a large repertoire of probes with orthogonal lifetime and spectrum signatures for high-plex target encoding (Aim 2), and (c) characterization, validation and benchmarking of Phasor S-FLIM for multiplexed spatial protein analysis using broadly relevant biological and clinical tissue models (Aim 3). Once developed, we expect the Phasor S-FLIM can detect at least 30 different protein targets through direct, one round of staining and imaging, in thick (>0.5 mm) tissues, with subcellular resolution (200 nm), and in high imaging throughput (1 x 1 mm2 plane in <15 min), which is currently not possible with existing methods. Upon successful completion of the proposed work, we will have established a working prototype ready to quickly serve the scientific community to address a broad range of biological and clinical questions that are previously impossible or impractical. Our technology can potentially shift current practice in interrogating protein and cellular processes as well as the complexity and systems in biology and disease with high resolution, throughput and scale.

NIH Research Projects · FY 2024 · 2022-09

Abstract Sulfur mustard (mustard gas) and arsenicals such as lewisite are major threats as chemical warfare or terrorist agents. Despite different chemical structures, both types of agents lead to similar skin damage with large blisters and wounds that heal slowly, raising the specter of shared pathogenic mechanisms. In this proposal, we will investigate the role of oxidant skin injury as a shared mechanism in mustard and arsenical vesicant injury, employing nitrogen mustard and phenylarsine oxide as surrogates for sulfur mustard and lewisite, respectively. Although major advances have been made in vesicant research, we still do not have effective counter agents. This may be in part due to lack of full understanding of the pathogenesis of vesicants. The overarching hypothesis of this proposal is that defining the early effects of vesicants in vivo at subcellular and single cell levels will refine our understanding of the role of oxidative stress in vesicant skin injury and lead to new therapeutic strategies. We will also test the hypothesis that cobinamide, a highly effective and versatile antioxidant that neutralizes both reactive oxygen and nitrogen species, will be effective as a counter-agent and as means to understand better the role of oxidative stress in nitrogen mustard- and phenylarsine oxide-induced vesicant injury. The specific aims are: 1) To understand the initiation of nitrogen mustard- and phenylarsine oxide-induced skin injury at a single cell level. We will use in vivo fluorescence lifetime imaging (FLIM) and single cell RNA sequencing to gain a new view on the initiation of vesicant injury and the role of oxidative stress in the pathogenesis. 2) To define the efficacy of cobinamide against nitrogen mustard- and phenylarsine oxide-induced vesicant skin injury. We will test the efficacy of cobinamide against nitrogen mustard- and phenylarsine oxide-induced vesicant skin injury in mice. Due to differences between mouse and human skin, we will also investigate vesicant mechanisms in transplanted human skin. These studies are significant and innovative because they use state of the art tools to investigate a gap in our knowledge about the role of oxidative stress in the initiation of mustard and arsenical vesicant injury and because they test for the first time cobinamide, a highly effective antioxidant that is bifunctional for reactive oxygen and nitrogen species.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Alzheimer's disease (AD) is the most common cause of dementia among elderly and is an apparent public health challenge in the U.S. as well as many other countries. Despite the extensive research effort on all aspects of AD, the exact causes of late-onset, sporadic AD remain elusive. Although genetic predispositions including known risk genes from human genetics studies are playing a prominent role in the pathogenesis and etiology of AD, recent growing bodies of evidence strongly suggest an emerging role of environmental contribution, particularly toxic constituents of air pollution including, but not limited to, particulate matter (PM) and metals, to the progression and onset of AD. Thus, it is critical to investigate how air pollution drives neurodegeneration and whether AD risk genes modulate its neurotoxicity and neuronal death, as an empirical example of the gene x environment (GxE) interactions to support the mechanism-based etiology of AD. As increasing number of studies in humans and animal models have confirmed the pivotal role of currently known risk genes and exposure to air pollution, independently or in combination, in AD neuropathology and neurodegeneration, we stay focuses on determining key neurotoxic mechanisms of ambient PM, the most abundant toxic constituents found in the ambient air, which subsequently leads to accelerated neurodegeneration and the clinical onset of AD. The overarching objectives of this study are 1) to comprehensively evaluate neurotoxicity of ambient PM and its environmental risk in a novel mouse model of late-onset AD, and 2) to identify key genes and mechanisms that determine the sensitivity or resilience to neuronal death triggered by PM. In this application, we include several innovative tools, such as a novel mouse model, human iPSC-derived neurons and the CRISPR-based functional genomics, to carefully assess the neurotoxicity of PM, its risk for developing and exacerbating AD phenotypes, and the GxE interactions, all of which could be more clinically relevant and applied to broader general population compared to existing findings from widely used animal models overexpressing familial AD mutations. In our knowledge, proposed in vivo and in vitro models are the best suited models to simulate and evaluate the environmental contribution to late-onset AD in humans. Thus, outcomes from this study maintain high translational significances to understand neurodegenerative mechanisms and the GxE etiology of late-onset AD. Lastly, this proposal is feasible, highly- significant, and highly-relevant to evaluate the etiology and progression of AD in the context of the GxE interactions. We believe that the outcomes could have large impact to the field, while accelerating progress towards understanding the environmental impact on the pathogenesis of AD.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Amyloid plaques are one of the canonical pathological hallmarks of Alzheimer's disease (AD). The amyloid hypothesis is a standard model for amyloid Abeta pathogenesis that has driven drug discovery for the past 25 years, giving rise to many clinical failures, including drugs that make the treated patients cognitively worse than the placebo-treated controls. Aducanumab (Aduhelm) is the first disease modifying treatment recently approved by the FDA on the basis of its ability to facilitate the removal of amyloid plaques, indicating the importance of these strictures. There are several different types of plaques and amyloid deposits known in AD, including diffuse, “classical” “dense core”, neuritic plaques and cerebrovascular amyloid (CVA) and intraneuronal amyloid deposits. While there is much that is known about amyloid plaque morphology and composition in AD, less is known about mechanisms of plaque deposition, their dynamics and interrelationships, the contributions of different cell types to their formation and their significance for AD pathogenesis. We seek to investigate these critical aspects of amyloid plaque deposition in a detailed and un unbiased fashion by labeling the proteome of specific cell types with the non-canonical amino acid, azidonorleucine, and following the incorporation of ANL-labeled proteins into amyloid deposits. The goal of this proposal is to determine the temporal, spatial and cellular dynamics of amyloid deposition using cutting-edge biorthogonal non-canonical amino acid tagging (BONCAT) technology and specific Cre driver mouse lines for cell specific synthesis of clickable proteins, exploiting click chemistry for fluorescence localization (FUNCAT), purification and enrichment and biochemical analysis. Our central hypothesis is that different types of plaques are deposited by different mechanisms at different times and locations by different populations neurons and that some of these types of plaques may be mechanistically unrelated to the other types of amyloid and may be differentially associated with pathogenesis and neuronal degeneration.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The overarching objectives and specific aims of this IPERT program are to (1) prepare trainees for successful multi-disciplinary Biophotonics careers and support biomedical workforce development. This will be achieved by (2) advancing the technical and operational proficiency of our trainees in Multiscale Biophotonics through an integration of modeling, computation, experiment, and problem-based learning, and (3) providing our trainees with sustained mentorship for professional development. This will be accomplished by establishing foundational technical and operational skills in the area of Multiscale Biophotonics during a 12-day onsite short course at the University of California, Irvine. This 12-day short course utilizes an integrated modular design and evidence based learning consisting of (a) lectures, (b) technology demonstrations, (c) laboratories combining hands-on data acquisition with computational analysis using our custom Virtual Tissue Simulator (VTS) software platform, (d) Team Science training, (e) mentored capstone problem-based learning (PBL) proposal development, (f) guest lectures on the use of an interdisciplinary Biophotonics approach to address biomedical problems, (g) panel discussions on academic and industrial career paths, and (h) tours of academic and industrial facilities. Further application and growth in trainee skill development will be accomplished by a mentored capstone problem-based learning project to be pursued for one year following the short course completion. In parallel with the short course and year-long PBL project, we will offer both technical and professional mentorship. Our technical mentorship aims to support Biophotonics, instrument and measurement design, technology development, as well as modeling, computation and usage of open-source Computational Biophotonics Tools. Our professional mentorship program aims to aid trainees in identifying appropriate short- and long-term professional goals and develop strategies to attain them. The course has been designed for interdisciplinary group of graduate student, post- doctoral researcher, and faculty scientists and engineers. We propose an aggressive outreach plan to support the participation of trainees who are traditionally underrepresented within STEM as well as from geographic regions that have historically received low levels of NIH support. We aim to train up to 20 individuals each year. In summary, our program provides a technical foundation and mentoring support for trainees to advance and apply an integrated technical proficiency in Multiscale Biophotonics to solve biomedical challenges. As the integration of expertise spanning modeling, computation and experiment are in demand in biomedical research and technology development, the proposed program advances NIGMS programmatic goals for training a diverse biomedical research workforce with expertise most relevant to the NIGMS Division of Biophysics, Biomedical Technology, and Computational Biosciences.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Distant-acting (“remote”) transcriptional enhancers bound by transcription factors (TFs) drive gene expression patterns in space and time. The importance of this function is underscored by mounting evidence linking mutations affecting remote enhancers to human disease. What remains unclear is how these remote enhancers regulate their target genes over long genomic distances, often skipping intervening genes. Remote enhancers typically interact with target promoters with the support of higher-order chromatin organization, but disruption of this chromatin organization does not abolish enhancer–promoter interactions. What additional factors regulate these long-range regulatory interactions remains largely unexplored. To address this central question, the proposed project will use a novel gain-of-function approach to identify cis-regulatory sequences that mediate long-range enhancer–promoter communication in the context of mammalian development. Traditional methods that assess in vivo enhancer activity rely on enhancer-reporter transgenesis; however, technical limitations preclude this approach from assessing long-range enhancer activity. The proposed work overcomes this limitation by utilizing efficient enhancer replacement technology in mice to systematically characterize in vivo long-range enhancer activity and identify cis-regulatory sequences that are critical for this long-range activity. To determine the mechanism of long-range regulation by these remote control elements and identify regulatory proteins that bind them, this work will use a multidisciplinary approach to map and visualize enhancer–promoter contacts at macromolecular resolution. Functional characterization of remote control elements will expand the repertoire of known cis-regulatory elements, provide a novel general mechanism for long-range enhancer– promoter communication, and substantially advance knowledge of the role of 3D-genome organization in the context of development and disease.

NIH Research Projects · FY 2025 · 2022-09

Abstract The goal of cancer immunotherapy is to build long-lasting tumor-specific immunologic ‘memory’ in patients that enables the lifelong rejection of tumors. The two prominent types of antigen-specific cancer immunotherapy, adoptive T cell therapy and APC-based vaccination, both require expansion of anti-tumor T cells via APCs. However, for the purpose of effective adoptive T cell therapy, the critical question is how to generate, within a short period of time, large numbers of antitumor T cells. Furthermore, in vitro-expanded T cells must also possess the capacity to engraft, proliferate, and persist in vivo with sufficient antitumor function to induce sustained antitumor activity. Autologous antigen-presenting cells (APCs) such as DCs also have several serious limitations. The necessity to access large amounts of cancer patients’ blood to prepare autologous APC from each patient in a timely manner is cumbersome. To overcome these problems, we developed the microfluidic process to generate cell-sized unilamellar vesicles (CUVs) and decorated them with antigen presenting ligands for artificial APCs (or aAPCs). Preliminary results show that aAPCs are able to bind and interact with T cells and cause their expansion. The objective of the present proposal is to further optimize the aAPCs preparation and test its capacity to induce tumor specific responses in vitro and in vivo. The hypothesis is that the optimized aAPC functionalization will result in enhanced expansion of cytotoxic CD8 T cells and a reduction in tumor progression over the present one (original). The Specific Aims are- 1) Bioinspired optimization of artificial antigen presenting cell (aAPC) production via microfluidic engineering. We will insert the antigen presenting ligands in the membrane to mimic cells. The aAPCs will also be produced with hydrogel cytoskeletons to optimize its mechanical properties for maximum T cell expansion. 2) Evaluation of the capacity of aAPCs to induce tumor specific T cell responses in vitro. Using PBMCs from healthy donors and breast cancer patients we will evaluate the capacity of aAPCs to induce cytotoxic T cells. 3) Evaluation of the capacity of aAPCs to induce T cell responses and tumor killing with an in vivo mice tumor model. Methods to scale up the production of aAPCs for in vivo use will be developed. The capacity of aAPCs to kill tumor in vivo in mice will also be determined using a melanoma model. The goal is to produce an aAPC preparation that mimics cells, is stable, easy to produce in large quantities and capable of expanding tumor specific CD8 T cells for immunotherapy of cancer.

NIH Research Projects · FY 2024 · 2022-09

Abstract Sacramento County, California, is one of the counties with the largest arrivals of Arabic-speaking refugees and Afghan and Iraqi special immigrant visa holders (SIVs) in the nation. Post-resettlement unemployment and poverty, poor English skills, socioeconomic pressures, cultural differences, lack of familiarity with preventive services, and inability to navigate the maze of the US health care system pose substantial barriers to refugee access to needed reproductive health (RH) care leading to disparities in maternal and infant health outcomes and to higher cervical cancer rates. An important factor to achieve health equity for refugees and SIVs is access to accurate and understandable information on sexual and reproductive health needs and services. In this 3-year study. With expert and outreach support from the Digital Scholarship Services of the University of California, Irvine (UCI) and the Sacramento Public Library, the California's Refugee Reproductive Health Network (ReproNet) will leverage its solidly established regional academic – refugee community partnerships to enhance Afghan and Arab refugee women's knowledge and access to accurate and linguistically appropriate RH information. Key ReproNet partners in this study will be the University of California, Davis and Muslim American Society Social Services Foundation (MAS-SSF). In this project, ReproNet will aim to: (1) Create a publicly-available digital repository of multilingual reproductive health resources (Dari, Pashto, Arabic) for Afghan and Arab refugee populations. Links to the materials of the repository will be posted on ReproNet’s social media pages and channels. The repository will be housed at the UCI DSS which will also maintain and store the resource materials beyond the project period. (2) Enhance Afghan and Arab refugee’s reproductive health literacy (RHL) in California through 12 in-person training sessions with the Sacramento Public Library and MAS-SSF (3 each in Dari, Pashto, Arabic and English) and 12 on-line training sessions offered via zoom or Facebook and (3) Increase the capacity of refugee providers to integrate reproductive health literacy in their programs. We will evaluate the project through google analytics, session evaluations, and pre- and post RHL and provider training assessments. An enhanced literacy and access to digital health information and library resources nationwide will strengthen refugee women’s capacity to obtain, process, and understand the basic RH health information needed to make optimal health decisions for themselves and their families.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Each step-change in increasing resolution on RNA identification and quantification (i.e., transcriptome profiling) has been transformative to our understanding of cell biology. As the most recent technological revolution, the ability to map the spatial distribution of RNA molecules has revealed the importance of subcellular RNA localization, advanced our understanding of cell and developmental biology, and transformed the single cell transcriptomics field. However, despite the recent development, current RNA-mapping tools are limited to molecules with long specific sequence (mostly > 500 nt). Many other important RNA species (such as splicing isoforms, miRNA, and RNA editing), with much shorter specific sequence motif, remain inaccessible in situ. Our goal is to make this next resolution jump in profiling transcriptomics, i.e., RNA-mapping with high transcript- selectivity and single base-sensitivity. We plan to achieve the goal by developing a new tool, named SMOOTHY- FISH (Single-nucleotide specificity, Minimum Of Off-Target effects, and High Yield), which enables spatially mapping all types of RNA molecules in individual cells. In this proposal, we will demonstrate SMOOTHY-FISH from two aspects: technique development and one example application test. More specifically, for the technical development, we anticipate achieving a platform that can suggest robust and reliable SMOOTHY-FISH probe- sets to any assigned RNA target sequence, a user-friendly system that will be widely accessible. To achieve the goal, we will incorporate the unique power of magnetic tweezers (a single molecule tool with nanometer and milliseconds resolution) to systemically quantify the dynamics and kinetics of nucleic acid hybridization under various conditions, to reveal how fast and how stable the designed SMOOTHY-FISH probe-sets can form the needed secondary structure, and to build a mechanistic modeling framework to describe and predict the hybridization dynamic process. As for the example application test, we will use SMOOTHY-FISH to study the mystery biogenesis process of circular RNA with isoform-specificity (i.e., co-existence with its linear isoforms) for the first time. We anticipate revealing the potential to modulate circular RNA expression quantitatively and efficiently, i.e., lay a foundation for therapeutic strategies in the future. Together, this work will provide a next- generation tool for in situ RNA identification and quantification, allow us to localize and quantify many important, but currently inaccessible RNA species at the single molecule level in individual cells, thus help open a new era of RNA biology, from basic science research to clinical applications.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Limited English Proficiency (LEP) Latinxs experience longer duration of untreated mental health disorders. In the case of depression and anxiety, Latinxs are half as likely as whites to receive quality, evidence-based care despite primary care providers recommending treatment at equivalent rates. There are known barriers for LEP Latinx patients with depression and anxiety, especially at the healthcare setting level. Because these patients are more likely to receive care in public healthcare settings which disproportionately care for patients on Medicaid or who are uninsured, there are often language and culturally concordant care models in these systems yet an overall deficiency of Spanish-speaking behavioral/mental health clinicians. In addition, these under-resourced healthcare systems also are less likely to be the site for implementation of innovations that might increase access to mental health care for their population. Digital interventions, i.e., those that leverage Internet and mobile technologies, can provide an effective option for overcoming these barriers. In many countries, digital treatments are frontline treatments for mental health issues such as depression and anxiety, creating a standard of care that can overcome disparities and promote health equity. In the United States, few patients have access to evidence-based digital treatments, and even when available, uptake and engagement is often low. This proposal aims to implement and evaluate an evidence-based digital cognitive-behavioral therapy intervention in safety-net primary care clinics for LEP Latinx patients with depression and/or anxiety. Primary care is the de facto treatment setting for the treatment of common mental health problems such as depression and anxiety and Latinx patients tend to prefer to remain treated in primary care rather than being referred to other specialty mental health services. We will conduct an effectiveness-implementation hybrid trial (Type 2) design with both provider- and patient-level randomization. This will generate data on effectiveness of digital peer support for mental health outcomes for LEP Latinx patients while simultaneously generating rigorous data on the implementation strategies with the highest chance of dissemination in future work. At the provider-level we will compare outreach (using our clinic patient registry) with inreach (traditional provider referral), at the patient-level we will compare two modes of delivery of the dCBT platform (peer-supported vs. unsupported). The long-term goal of this research is to aid in the implementation of digital mental health interventions that can be sustainable implemented in low-resourced settings, while reducing the reliance on professionals, overcoming workforce deficits, and increasing relevance for diverse populations. Such interventions could be provided at scale and address the substantial burden of disease resulting from common mental disorders while making resources more available to those traditionally underserved by our health system.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT This P30 application seeks support for the Cores of Vision Research within the University of California Irvine (UCI) that will strengthen and enhance NEI R01-funded research. The project also aims to support the major expansion of vision research currently under way at UCI. It will broaden and optimize research capabilities, support and expand existing research projects, and help establish new studies, ultimately boosting productivity and strengthening vision research at UCI. Three research Core modules have been identified that would provide substantial benefit to current and future NEI R01-funded projects at UCI: (i). Ocular Microanatomy; (ii) Visual Function and Non-invasive Ocular Imaging; and (iii) Ocular Mass Spectrometry, Lipidomics, and Proteomics. In addition, an Administrative Group will be established to facilitate and support the function of these research Cores. The three research Cores were selected by majority votes of all current NEI R01-funded investigators, and other investigators who are directly or indirectly involved in ophthalmic research at UCI. The most requested services were developed into the Cores described in this P30 application. The vision research community at UCI has grown significantly over the past several years with the recruitment of 4 senior and 2 junior investigators and the formation of the Center for Translational Vision Research (CTVR), with at least two more mid-career faculty recruitments in the works. This expansion has been made possible by the exceptional institutional support from the Department of Ophthalmology and the School of Medicine, who have committed significant financial and infrastructure resources to bolster the vision research community at UCI. The services provided by the Cores for Vision Research will meet the scientific needs of the NEI R01-funded researchers as well as of early-stage investigators, and will increase efficiency, accelerate progress, and encourage new research directions and scientific collaborations at UCI. The Cores will also provide training and access to state-of-the-art experimental tools for students and postdoctoral scholars and will help prepare them for successful careers in ocular research.

NIH Research Projects · FY 2025 · 2022-09

Abstract Primary Open Angle Glaucoma (POAG) is the most common form of glaucoma that leads to irreversible vision loss. Elevated intraocular pressure (IOP) due to dysfunction of trabecular meshwork (TM) tissue is a hallmark of POAG. However, the pathological mechanisms leading to TM dysfunction and IOP elevation are poorly understood. TM has an intrinsic ability to maintain IOP homeostasis by sensing the changes in the flow of aqueous humor (AH). In this regard, we recently showed that Ca2+ influx through transient receptor potential vanilloid 4 ion channels in the TM (TRPV4TM channels) lowers IOP via activation of endothelial nitric oxide synthase (eNOS)–NO signaling. Importantly, we showed that TRPV4TM-eNOSTM signaling is impaired in glaucomatous primary human TM cells. The major goals of this application are to elucidate the pathological mechanisms that impair TRPV4TM-eNOSTM signaling in glaucoma and to target them for rescuing the TM function. Chronic endoplasmic reticulum (ER) stress is a crucial contributor to TM dysfunction and IOP elevation in glaucoma. In our preliminary studies, we observed that chronic ER stress activates inducible NOS (iNOS), an enzyme commonly associated with the formation of oxidant molecule peroxynitrite (PN). PN levels are elevated in TM tissues from POAG donor eyes and exogenous PN reduced TRPV4TM channel activity in human primary TM cells. Moreover, induction of ER stress also lowered TRPV4TM channel activity. Therefore, we hypothesize that PN-induced inhibition of TRPV4TM-eNOSTM signaling contributes to TM dysfunction and IOP elevation in glaucoma. The major objectives of this application are to determine whether chronic ER stress leads to TM dysfunction and IOP elevation via PN-induced lowering of TRPV4TM-eNOSTM signaling in glaucoma, and to target this pathology for the treatment of glaucoma. In Aim 1, we will determine whether chronic ER stress lowers TRPV4 channel activity in TM cells. In Aim 2, we will determine whether PN levels are elevated in glaucoma and whether PN lowers TRPV4 channel activity in TM. We will also determine whether chronic ER stress underlies elevated PN levels in glaucoma. In Aim 3, we will target PN pathology to lower elevated IOP in mouse and human models of ocular hypertension. This proposal utilizes complementary expertise of Dr. Zode’s laboratory in glaucoma research and ER stress, and Dr. Sonkusare’s laboratory in TRP ion channel imaging and electrophysiology. This study will utilize state-of-art Ca2+ imaging, patch-clamp, eNOS activity, and nitric oxide measurements in primary human TM cells and TM tissues, human perfusion cultured donor eyes, and mouse models of glaucoma. Successful completion of the proposed studies will provide novel pathological mechanisms and therapeutic targets for the treatment of general POAG.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Regulator of G protein Signaling (RGS) proteins play a key modulatory role in G Protein-coupled receptor (GPCR) signaling. Through both G protein-dependent and -independent mechanisms, RGS proteins play important roles in disease, and this has driven numerous efforts to pharmacologically target their function. These efforts are, however, hindered by the fact that RGS proteins are considered difficult drug targets, and identifying mechanisms that control RGS protein activity, expression and/or subcellular localization has therefore become an important area of investigation. Our long-term goal is to determine how levels and activity of RGS proteins is regulated, with a view of identifying druggable “soft-spots” within the regulatory network. Our rationale for the current proposal is that identification of specific mechanisms regulating RGS2 will uncover therapeutic points of intervention to increase RGS protein levels and activity. Low RGS2 protein levels or activity are associated with a wide range of pathologies, including hypertension, heart failure and asthma, and our central hypothesis is that pharmacologically enhancing RGS2 protein levels would have broad clinical implications. Our objective in this proposal is to decipher posttranslational mechanisms regulating expression, activity and subcellular localization of RGS2. We will focus on two crucial, but understudied, mechanisms regulating RGS2 protein levels and activity; proteasomal degradation and phosphorylation. We identified a Cullin-RING E3 ligase (CRL) targeting RGS2 for proteasomal degradation, and recently identified the degron in RGS2 that is recognized by F-box only protein 44 (FBXO44), the substrate-recognizing component of the CRL. In Aim 1 we will use biochemical and structural approaches to determine how FBXO44 interacts with RGS2. We will also determine whether FBXO44-RGS2 binding is determined by the associated CRL. FBXO44 degrades RGS2 only in the context of a CUL4B/DDB1, but not a CUL1/Skp1 complex in cells suggesting that FBXO44 RGS2 specificity may depend on the nature of the CRL. In Aim 2 we will determine the role of phosphorylation for RGS2 function. Phosphorylation plays a central role in signal transduction cascades, however there is a lack of comprehensive information on the global role of phosphorylation for RGS2 protein function. Our previous studies identified potential importance for PKC and Src kinase in regulating RGS2. We will determine which residues are phosphorylated, as well as the consequence for RGS2 protein stability, using a wide range of techniques, including PhosTag gel electrophoresis, in vitro kinase activity assays, as well as LC-MS. In Aim 3 we will determine the functional consequences of altered posttranslational RGS2 regulation. We will determine effects on G protein-dependent and -independent RGS2 functions in both transfected cells, physiologically relevant cell lines and ex vivo models. The expected outcome of these studies will be a detailed view of how RGS2 protein levels, activity and subcellular localization is regulated by posttranslational mechanisms. Through these efforts, we will make important inroads to future drug discovery efforts targeting RGS2.

NIH Research Projects · FY 2025 · 2022-09

Project Summary As the number of Americans living with Alzheimer’s disease (AD) is projected to reach 13 million by 2050, we must prioritize efforts for early disease detection. The bottleneck of early AD detection has greatly hindered clinical treatment and development of successful therapeutics. With greater availability of multimodal AD biomarkers in clinical practice, we have a unique opportunity to leverage statistical machine learning for earlier detection of AD. Past work has demonstrated good classification accuracy of clinical diagnosis of AD using binary classifications (i.e., AD-dementia vs healthy cognition, HC; mild cognitive impairment, MCI vs HC; AD vs MCI). These classifiers, however, are often reliant on unimodal biomarker inputs, and no formal comparison of multimodal biomarker integration (“fusion”) methods exist for predicting either AD clinical diagnosis or biomarker status as defined by the A/T(N) framework. Lack of optimal multimodal fusion strategies and holistic diagnosis prediction beyond binary classification reduce the translational value of statistical machine learning classifiers in clinical practice. This proposal fills these gaps by evaluating several competing strategies for multimodal fusion and multiclass classification (e.g., AD vs MCI vs HC) using data from the National Alzheimer’s Coordinating Center and Alzheimer’s Disease Neuroimaging Initiative. The strength of using large, multimodal datasets for disease prediction is accompanied by the challenge of handling missing data, a barrier for building a reliable classifier. This proposal will address these challenges with two specific aims: (1) compare techniques for optimal data imputation and multimodal fusion, and (2) develop a multiclass model to accurately predict AD status (AD/MCI/HC and A+T+/A+T-/A-T-) using multimodal inputs. Preliminary analyses of multimodal data fusion in binary classification using random forest and sparse group lasso classifiers motivate Aim 1. Preliminary analysis of two strategies of multiclass classification demonstrates feasibility of developing a multimodal, multiclass classifier for Aim 2. The proposed work will be enhanced by the excellent training and research environment at the University of California, Irvine (UCI), including direct access to 1 of 33 NIA-funded Alzheimer’s Disease Research Centers (ADRCs). The ADRC offers a third, independent dataset to serve as a validation set to improve the rigor of results from the proposed experiments. The applicant will be supported by the joint mentorship of Dr. Craig Stark, the ADRC Biomarker Core Leader, and Dr. Babak Shahbaba, Director of the UCI Data Science Initiative, and will receive advanced training in both aging and AD research and statistics and machine learning techniques. Fellowship training will be further strengthened by the additional mentorship of Dr. Peter Chang for machine learning, Dr. Michele Guindani for multimodal data fusion, and Dr. S. Ahmad Sajjadi for clinical expertise in AD. The proposed training and research plans will result in the development of a reliable, multimodal, and multiclass classifier for AD status prediction to enable earlier disease detection and stratification of patients for more effective clinical trials.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Half of the known human genetic variations that contribute to disease are due to single nucleotide polymorphisms (SNPs). Thus, there is a pressing need to develop precision genome editing tools that are able to correct these SNPs with high efficiency and accuracy. Current CRISPR-Cas based precision genome editing tools, such as DNA base editors and prime editors, were designed to perform targeted single nucleotide changes without introducing double stranded breaks and relying on the homology-directed repair pathway. However, these tools have several drawbacks observed in cells, such as off-target DNA and RNA editing, low efficiency, and unintended editing of nucleotides within the neighborhood of the target nucleotide (bystander editing) leading to undesired genomic changes. Moreover, DNA base editors are able to perform only transitions (interchanging purines (AG) or pyrimidines (CT)) but not transversions (interchanging pyrimidines for purines and vice versa). These shortcomings reduce the targeting capabilities of current precision genome editing tools and are the key limitations of using them as therapeutic agents. Building on our recent work that explains the molecular basis of the DNA base editors’ drawbacks, we propose four innovative strategies to design precision genome editing approaches that address the limitations of current genome editing tools and expand their targeting scope. Three of the four strategies will yield base editors with dual programmability. Besides the programable nuclease (Cas9) that guides base editors to the sequence of interest, these novel base editors will additionally have easily programmable catalytic modules that will allow selecting only one nucleotide for editing. This dual programmability will eliminate the bystander editing and make these DNA base editors exceptionally accurate. Two out of four designs will yield DNA base editors able to perform transversions (interchanging pyrimidines for purines (CG and TG)) and to correct additional ~25% of pathogenic SNPs inaccessible by current base editors. Moreover, one of the two transversion base editors will possess dual programmability, hence will be exceptionally accurate. Overall, the four strategies proposed here will yield the next generation precision genome editing tools that, besides the direct therapeutic corrections of SNPs’ in vivo, will also allow interrogating the association between multiple SNPs, gene expression and diseases (neurodegenerative diseases or various types of cancers). Thus, these DNA editing tools will pave the way for investigating the molecular mechanisms of multiple genetic disorders and enable us to develop new therapeutic strategies.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Early detection of melanoma is a key factor in improving patient survival and decreasing treatment costs. The sensitivity of dermoscopy, the standard of care in the diagnosis of melanocytic lesions, was reported to be highly variable, ranging between 68-96%, depending on the proficiency of the physician and the stage of the lesion. Low sensitivity reflects high rates of false-negative findings, which delay diagnosis and treatment. Thus doctors must err on the side of caution, which leads to an excess of unnecessary biopsies and increased medical costs. Distinguishing cutaneous melanoma from benign melanocytic nevi with high accuracy based on dermoscopy remains a challenge even when in the hands of expert clinicians since this approach only offers a two-dimensional image of the lesion's superficial structure. Ultimately, a biopsy is necessary for definitive diagnosis by the dermatopathologist, but this too may be affected by inter-observer variability, resulting in discordant conclusions. A study performed at the Melanoma Center, at UCSF estimated that 214,500 to 643,500 cases of melanocytic neoplasms in the United States would be diagnosed differently by another dermatopathologist, annually, which has significant consequences for the patient regardless of the nature of the lesion. We propose to develop and clinically evaluate a fast, large area multiphoton exoscope (FLAME) as a tool for non-invasive imaging and early detection of melanoma in order to reduce false positives and false negatives in both dermoscopy and histopathology. Multiphoton microscopy (MPM) is a nonlinear optical imaging technique that provides unique structural and molecular contrast based on endogenous signals such as second harmonic generation from collagen and two-photon excited fluorescence from NAD(P)H/FAD+, keratin, melanin and elastin fibers. In preliminary studies, we demonstrated that macroscopic areas of skin (cm2 scale) could be mapped out with microscopic resolution within ~2 minutes by combining optical and mechanical scanning mechanisms with deep learning image restoration. As required by PAR-20-155 our academic-industrial partnership will deliver a powerful MPM imaging tool to clinicians for non-invasive, real- time quantitative assessment at the bedside that would not require specialized training. Our proposed application is for early diagnosis of melanoma, but the approach will have wider impact, for rapid, in vivo characterization of cellular morphologic and metabolic imaging endpoints in patients. Our specifics aims are: (1) to develop FLAME, a compact, portable MPM prototype system for rapid, depth-resolved in vivo imaging of skin, over macroscopic areas (cm2-scale) with microscopic resolution and enhanced molecular contrast; (2) to implement safety features and demonstrate the technical feasibility; (3) to test the performance of FLAME by evaluating its ability to provide in vivo quantitative optical endpoints with sufficiently high predictive power to reliably distinguish benign from early melanoma lesions. We are a multi-disciplinary team of investigators from UC Irvine, Vidrio Technologies, LLC and Tufts University with 3 to 8 years record of collaboration.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY In eukaryotes, transcriptional regulation is essential to maintaining cell identity, responding to intra- and extra-cellular signals, and coordinating gene activities, whereas its dysregulation can cause a broad range of disorders. Previous methods mainly used averaged genomic signals from thousands of cells to investigate gene regulation, failing to reveal the regulatory heterogeneity across diverse cell states. Recent advances in multimodal single-cell technologies provide new opportunities to decipher the cell-type-specific regulation code at the finest resolution possible. However, its computational modeling is still in its infancy due to the high dimensionality, missingness, vulnerabilities to confounding factors, and complex feature interactions. In this project, we aim to develop a suite of computational models to construct gene-centric, personal regulome via single-cell multiome integration and link multi-scale dysregulations to disease. Distinct from previous efforts reporting a set of one-dimensional (1D) functional cis-regulatory elements (CREs) from only one genome and applying it to all samples, we aim to construct personal, compact, gene-centric, and cell-type-specific transcriptional regulome from sc- multiome data. Specifically, we will first propose a scalable multimodal deep generative model to integrate single-cell data with single-, multi-, and hybrid modalities. Distinct to existing methods, we will include an invariant representation learning scheme to derive latent cell representations uncorrelated with confounding factors (e.g., age, gender, read depth, and batch effects) for bias- free transcriptome and epigenome reconstruction (Aim 1). Then, we will go beyond the 1D genome annotation by deciphering multi-scale gene regulation code (Aim 2), including i) functional CREs at a base-pair resolution; ii) CRE target genes for functional interpretation; iii) transcription factor regulatory networks. Lastly, we will develop interpretable deep learning models to link multi-scale dysregulations to disease with mechanistic explanation (Aim 3). This proposal is built on a close and long-term collaboration between Dr. Jing Zhang, an expert in computational biology and machine learning at the University of California, Irvine, with Dr. Feng Yue, an expert in regulatory genomics and 3D genome organization at the Northwestern University. Upon completion, our proposed methods will substantially deepen our understanding of transcriptional regulation to a single-cell level resolution and quantitatively relate multi-scale risk factors to genetic disorders. In addition, our aims will yield open-source software for the scientific community as essential tools for single-cell multi-omics data processing and integration.

NIH Research Projects · FY 2024 · 2022-09

Surgical disparities for adults and children have been identified as a major problem in the US and can be experienced at multiple points along a patient's health care trajectory. Data from our center indicates that a substantial portion of Latin American origin children who undergo surgery experience high anxiety and postoperative pain as well as postoperative impairments in psychological and physical functioning as compared to White non-Latin American origin children who undergo surgery. Since Over 60 million U.S. residents are of Latin American origin and 25.6% are children under the age of 16, this is a significant issue. Recent growth in use of mobile devices provides us an opportunity to create low- cost mHealth behavioral interventions to reduce this disparity in surgical outcomes. In a previous NIH award, the PI developed and tested an evidence based mHealth tailored intervention (WebTIPS) that aims to prepare and be a companion of a child and their family during a surgical event. WebTIPS aims to enhance the recovery of the child in several ways such as reducing anxiety and pain and is based on information provision, modeling, and teaching of coping skills. WebTIPS, however, was developed and validated with a population of primarily English-speaking children and their parents and hence the need to revise the intervention so it could be directed at other populations. Over the past 4-years, we have established multiple academic and community collaborations, conducted extensive participatory research with Latin American origin stakeholders and used the heuristic framework and a modified ecological validity model to culturally adapt WebTIPS. The culturally adapted intervention is called L-WebTIPS. The first phase of this application (R61) includes web programming of L-WebTIPS, conduct formative evaluation and conduct feasibly RCT to test this intervention. The second phase (R33) includes a multi-center RCT which aims to determine the effectiveness of L-WebTIPS compared to attention control intervention in decreasing postoperative pain, opioids consumption and lowering anxiety in Latin American origin children undergoing outpatient surgery. Secondary aims of the R33 include examining the impact of L-WebTIPS on home-based clinical recovery parameters such as pain, analgesic requirements, new onset behavioral changes and return to normal daily activity in Latin American origin children undergoing outpatient surgery. Finally we plan to determine if the use of L-WebTIPS reduces anxiety and enhance experience among the parents of Latin American origin children undergoing surgery. We submit that using a cultural adaption process for an existing validated intervention will accelerate the process of reducing surgical disparities and bringing an effective intervention to clinical settings and routine use

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Proper functioning of the human body relies on the organization of cells in 3D space. A cell’s function and fate are determined by its biomolecule composition and its 3D environment. The spatially identification of proteins, RNAs, and DNAs in a tissue thus provides a powerful map to decipher how cells build tissues and become diseased. Through the use of single-cell omics, it’s been possible to reveal rare cell types that benchmark development, oncogenesis, and brain functions. However, the cell isolation process in single-cell analysis unavoidably causes loss of spatial information. To obtain spatial information, spatial transcriptomics based on imaging or sequencing have emerged to give insight into the heterogeneous expression patterns in tumors, brain, and wound tissues. Unfortunately, most spatial transcriptomics methods can only examine thin tissue sections, and are incompatible with proteomics. To address these drawbacks, the goal of the project is to develop a conceptually novel 3D spatial multiomics technology featuring gel-based optical isolation (GO3D). The proposed GO3D technology is distinct from all current spatial omics, and will enable the profiling of proteins, RNAs, and DNAs of whole-mount tissues with subcellular resolution, high coverage and high throughput, simultaneously. This innovative design is based on the gel-based label-retention expansion microscopy (LR- ExM) that the PI published recently. GO3D will drastically transform our understanding of many critical biomedical questions, which we lack of tools to address currently. For example, how are cells in a highly dynamic skin migrate in 3D to heal wounds? How do specialized neurons build brain? And where do microbes interact with what cell types in gut?

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Close to 40% of home health patients have been identified as persons with Alzheimer’s Disease or Related Dementias (ADRD). As the prevalence of ADRD in the population in general is expected to double by 2050, so will the percent of patients with ADRD treated by home health agencies. During the Covid-19 pandemic many home health agencies have begun using telehealth technologies, including virtual visits and biomonitoring, to augment and partially substitute in-person, traditional care. This is an acceleration of a trend that began over a decade earlier. A survey has found that by July of 2020, 49% of agencies used some form of telehealth. These two emerging trends, the expected increase in patients with ADRD on the one hand and the increased penetration and utilization of TH by HHAs on the other, raise questions about whether the two are compatible. Can home health care provided via telehealth to patients with ADRD be as good as the care provided to these patients in person? Would it be comparable in terms of patients’ health outcomes and patients’ experience? These questions have not been addressed to-date. Furthermore, there is no information about the type of agencies that adopted telehealth, and in particular among those agencies that care for a majority of ADRD patients. There is also no information about the differences in types of telehealth technologies (e.g. communication telehealth versus biomonitoring) that were adopted. This study will address these questions by: 1.) performing a national survey of home health agencies caring for a majority of patients with ADRD about their telehealth capabilities and use, and the timing of telehealth adoption; 2.) linking survey data to agency characteristics, patients’ health outcomes and patients’ experience data; 3.) analyzing the data statistically to identify agency characteristics associated with different stages of telehealth adoption, the association between telehealth and better patients’ health outcomes and experiences, and the relative ranking of specific telehealth technologies in terms of their association with better patients’ health outcomes and better patients experiences. The information gained in this study will inform home health patients and families, advocates, the industry and CMS, as the issue of payment for telehealth services for home health is moving forward towards consideration by Congress and will influence federal and state policies in the coming years.