University Of California-Irvine

NIH Research Projects · FY 2025 · 2016-03

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. Our recent studies have shown that chronic endoplasmic reticulum (ER) stress is associated with the pathophysiology of glaucomatous TM damage and IOP elevation. However, the exact mechanisms of TM cell dysfunction/loss are not completely understood. The ER and mitochondria communicate constantly via mitochondria-associated ER membranes (MAMs) to regulate vital cellular functions including autophagy. Autophagy degrades long-lived proteins and damaged organelles including mitochondria (known as mitophagy) via lysosomes. Impaired mitophagy is known to cause abnormal accumulation of damaged mitochondria resulting into cell death. In our preliminary studies, primary human TM cells exhibited an abundant mitochondria and MAMs. Interestingly, human primary TM cells and TM tissues from POAG donor eyes demonstrated increased accumulation of mitochondria. Moreover, chronic ER stress-induced transcriptional factors, ATF4 and CHOP led to increased reactive oxygen species and impaired mitophagy in primary human TM cells. Our overall goals are to define the role of MAMs and impaired mitophagy in TM dysfunction and IOP elevation in POAG and to further target these pathways for the treatment of glaucoma. We hypothesize that chronic ER stress induces impaired mitophagy and mitochondrial dysfunction, leading to TM dysfunction/loss and IOP elevation in POAG. We will determine whether impaired mitophagy and mitochondrial dysfunction are associated with TM dysfunction and IOP elevation in human and mouse glaucoma (Aim 1). We will further determine whether chronic ER stress induces impaired mitophagy and mitochondrial dysfunction, leading to TM dysfunction/loss and IOP elevation (Aim 2). Finally, we will perform proof-of-principle studies exploring whether the mitophagy enhancers improve outflow facility and reduce elevated IOP in mouse models of glaucoma (Aim 3). We will utilize human primary TM cells and post-mortem TM tissues from normal and glaucoma donor eyes, mouse models of glaucoma and mitophagy flux reporter mouse model (mito-qc) as well as transmission electron microscopy (TEM) and the Seahorse assays to determine the role of MAMs, mitophagy and mitochondrial dysfunction in TM function and IOP homeostasis. The successful completion of the proposed studies will provide novel crosstalk between ER stress and mitophagy and target the pathological mechanisms for the treatment of general POAG.

NIH Research Projects · FY 2026 · 2016-03

Project Summary/Abstract Tendons and ligaments are fundamental components of a functional musculoskeletal system. Tendons attach muscles to bone and interact with these tissues at distinct attachment sites called entheses (bone) and myotendinous junctions (MTJs, muscle). These interactions cause unique changes in gene expression and extracellular matrix (ECM) production that allow tendons to bear the forces exerted by muscle contraction. Transcription factors such as Scleraxis (Scx) specify early tendon progenitor cells (TPCs) and regulate ECM production. We previously discovered a critical ECM scaffolding protein called Thrombospondin-4b (Tsp4b) in zebrafish that is regulated by Scx, required for tendon maintenance and conserved in human tendons. How different types of tendon fibroblasts (tenocytes) are specified and influence ECM assembly at entheses or MTJs remains unclear. The current proposal addresses these issues using the advantages of the zebrafish for single cell RNA sequencing, in vivo imaging and genetic manipulation. The long-term goal of the proposed research is to understand the spatial dynamics of gene regulation and ECM assembly in tendons/ligaments and their regulation by mechanical force. Three primary hypotheses guide the research: 1) distinct subtypes of TPCs develop at entheses and MTJs in response to force, 2) ECM secreted by tenocytes regulates force-dependent signals that alter these distinct modes of gene expression in tenocytes and 3) retinoic acid is a novel force-dependent signal controlling tendon development. Aim 1 will perform scRNA-seq in tenocytes and see how force alters gene expression profiles. Aim 2 will study roles for Tsp4b, ECM, and TGF-beta signaling in force-dependent gene expression in tendons. Aim 3 will study the roles of RA signaling in tendons, and its responses to mechanical load. Each aim combines novel single cell approaches, genetic manipulation, live imaging and quantitative methods for physiological stimulation of muscles to get at mechanisms of tendon cell specification and ECM assembly in response to force.

NIH Research Projects · FY 2025 · 2016-02

Abstract Transplantation of neural progenitor cells has extraordinary potential for the treatment of many nervous system disorders. Several recent studies have shown that progenitor cells derived from the embryonic mouse medial ganglionic eminence (MGE) retain a unique ability to migrate and differentiate into GABAergic interneurons following transplantation into the juvenile or adult rodent brain. We recently demonstrated that these cells are effective in correcting memory impairments and preventing spontaneous seizures in a mouse model of traumatic brain injury. However, it is not known whether similar therapeutic effects can be achieved with clinically-relevant cell sources, such as human pluripotent stem cells. Here, we propose studies to evaluate the effect of human-derived interneurons in a pre-clinical model of traumatic brain injury. Our approach involves transplantation of human interneuron progenitors into a widely-used rodent model of closed-head injury at different stages following injury followed by in vitro patch-clamp recordings, immunofluorescence techniques and neural circuit mapping to evaluate the synaptic integration of grafted neurons. A battery of behavioral assays and video-EEG monitoring will also be applied. Two specific aims are proposed: (i) determine how human-derived GABA progenitors integrate into brain injured hippocampus, and (ii) test the therapeutic efficacy of human GABA neurons in a mouse model of traumatic brain injury. If successful, our results will help move these exciting technologies closer to the clinic by establishing relatively direct proof of concept for human interneuron transplantation to treat traumatic brain injury.

NIH Research Projects · FY 2025 · 2015-12

Project Summary / Abstract Alternative polyadenylation (APA) plays an important role in the post-transcriptional regulation of most human genes. The broad importance of APA is well exemplified by the altered expression of NUDT21, a key APA regulator that we reported in the first cycle of this grant, in diseases such as glioblastoma, diopathic pulmonary fibrosis, and neuropsychiatric disorders. More recently, our work has revealed a novel mechanism by which 3ʹ- UTR shortening can repress tumor suppressor genes (e.g., PTEN) in trans by disrupting competing endogenous RNA (ceRNA) crosstalk, rather than by inducing oncogenes in cis. Aside from these few examples, the prevalence and functions of APA in a wide spectrum of human traits and diseases remain largely unknown. Most human traits/diseases have been found as associated with hundreds of thousands of noncoding single-nucleotide polymorphisms (SNPs) in numerous genome-wide association studies (GWASs). However, functional interpretation of these SNPs remains a significant challenge because GWAS data do not show how the SNPs work. To better understand their effects, expression quantitative trait loci (eQTLs) have been widely used to link GWAS SNPs to gene expression. Despite massive efforts on elucidating eQTLs, the functions of many GWAS SNPs remain unexplained. An important reason is that eQTLs do not consider APA regulation. We have recently constructed the first human 3′UTR APA quantitative trait loci (3′aQTLs), which contain ~0.4 million SNPs associated with APA of target genes, using ~8,000 GTEx v7 RNA-seq samples across 46 tissue types (Nature Genetics, accepted in 2021). These 3′aQTLs can explain ~16.1% of GWAS SNPs in 15 common traits/diseases, and they are largely distinct from eQTLs and splicing QTLs. Based on these exciting preliminary data, we hypothesize that computational and experimental modeling of APA will substantially facilitate the interpretation of numerous GWAS SNPs important for APA regulation, which are enriched in 3′UTRs and gene downstream regions. Hence, we propose to develop innovative bioinformatics and experimental methods for identifying 3′aQTLs and nominating APA-linked disease/trait susceptibility genes in a wide variety of cell types and environmental factors, followed by in vivo functional characterization using our unique CRISPR engineering system. We expect to establish APA as an emerging and important molecular phenotype to explain a large fraction of GWAS risk SNPs, leading to significant novel biological insights into the genetic basis of APA and APA-linked susceptibility genes in a wide spectrum of human traits and diseases.

NIH Research Projects · FY 2025 · 2014-02

Abstract: Our research focuses on microglia, a type of brain cell crucial to understanding and treating several central nervous system diseases. In our first funding cycle (2014-2019), we developed a technique to remove and replace microglia using a drug that targets a specific receptor (CSF1R), which has since become a standard method in neuroscience. This approach has significantly advanced our understanding of microglia and has shown therapeutic potential in various disease and injury models. In our second funding cycle (2019-2024), we further explored microglia replacement and discovered several key findings, including a new method of reprogramming microglia and a novel route for bone marrow-derived cells to enter the brain allowing us to fully replace the microglial tissue with bone-marrow derived monocytes/myeloid cells. Building on these findings, we propose three specific aims for our next research phase. First, we will investigate a new method of microglia recovery that avoids the need for complete microglia removal. This involves temporarily inhibiting CSF1R, leading to the reprogramming and proliferation of microglia and other brain cells. Second, we will explore the potential of replacing dysfunctional microglia with functional bone marrow-derived cells as a treatment for genetic disorders associated with microglia dysfunction. Lastly, we will study the role of peripheral myeloid cells entering the brain in the development of Alzheimer's disease. Our research aims to deepen our understanding of microglia biology and explore its therapeutic potential, moving towards more clinically relevant applications.

NIH Research Projects · FY 2026 · 2013-09

Studies in animal models linked gestational exposures to endocrine disrupting chemicals (EDCs) with the onset of disease in exposed and unexposed descendants. Many groups found such transgenerational effects of chemical exposures, which were proposed to be examples of epigenetic inheritance. Transgenerational effects of environmental exposures have substantial support in the literature. Yet the concept that responses to environmental exposures can be transmitted to subsequent generations through the germline without DNA mutations remains controversial because the underlying mechanisms have not been explained satisfactorily. Understanding how effects of environmental exposures are transmitted to unexposed generations without DNA mutations is a fundamental, unanswered question in biology. We developed a highly reproducible animal model for transgenerational inheritance of obesity. When pregnant F0 mouse dams were treated with environmentally-relevant (nM) doses of TBT via their drinking water throughout gestation, increased fat accumulation was detected in F1-F4 generation male descendants. Affected TBT-group males developed a transgenerational “thrifty phenotype”: they were resistant to fat loss during fasting, rapidly gained weight when dietary fat was increased and retained this fat even after being returned to a normal, low-fat diet. Our published and preliminary results led us to propose a new model for transgenerational inheritance - that prenatal TBT exposure altered higher-order chromatin structure (HOCS), changing secondary epigenetic modifiers that inhibited expression of insulin degrading enzyme (Ide) causing diet-induced hyperinsulinemia and obesity. Here we propose a comprehensive series of experiments designed to determine exactly how exposure of pregnant F0 dams to TBT alters HOCS in F1-F3 primordial germ cells (PGCs), why these changes are inherited, rather than reversed to the normal state, how these changes affect lower-level epigenetic regulators controlling expression of Ide and why does the phenotype only occur in males. Aim 1 will identify mechanisms that drive changes HOCS near the Ide gene and how these interact with lower-level epigenetic regulators to modulate Ide expression in the adult liver. Aim 2 tests whether epigenetic interventions, such as dissolving the HOCS alterations or releasing Ide expression from repression in PGCs or adults can prevent or reverse the transgenerational predisposition to male-specific metabolic phenotypes. Deciphering the underlying mechanisms will have profound implications for how the field views transgenerational inheritance and how future experiments are planned and conducted. This new understanding will be critical to explaining the etiology of non-communicable diseases such as obesity and type 2 diabetes, targeting their causes and ameliorating their effects. Our results will have broad implications for understanding epigenetic transmission of the effects of environmental stressors and could offer opportunities to incorporate considering prevention of transgenerational inheritance into risk assessment paradigms.

NIH Research Projects · FY 2025 · 2013-07

Enter the text here that is the new abstract information for your application. In 2013, we launched our Training Program in Stem Cell Translational Medicine for Neurological Disorders (“training program”) at the University of California, Irvine (UCI) with the premise that such training is essential if stem cells are to be used successfully to treat neurological disorders. Nine years later, we believe that this premise is even more true, as many more academic groups and biotechnology companies explore the potential of stem cells to directly treat, or lead to treatments for, human disease and injury. Three main area of translational research are being conducted: the use of stem cells or their derivatives for transplantation to replace those cells damaged or destroyed by injury, the use of stem cells, including patient derived cells, to study disease processes and mechanisms, and the use of differentiated products of stem cells to screen for drugs that could alleviate disease or damage. In recent years the addition of CRISPR-mediated gene editing and the development of “brain organoids” have added new tools to the arsenal of researchers in the field. For the promise of stem cell treatments to become a reality, not only must basic research advance in this rapidly evolving field, but these advances must also be translated through preclinical and clinical development into clinical practice. Translational approaches must be taught directly. The goal of this renewal is to provide an even better environment to train a new generation of scientists in the translational application of stem cell biology to neuroscience. Building upon key successes and feedback from the trainees over the course of this T32 since its inception, our updated program will even better serve this purpose and includes (1) state-of-the-art rigorous and reproducible wet lab science employing training in the fundamental principles of rigor and reproducibility, as well as quantitative and statistical methods; (2) clinical experiences to understand and plan research efforts in the context of clinical translation; (3) knowledge regarding practical aspects of moving discoveries to clinical trials and therapies through industry internships and Sue and Bill Gross Stem Cell Research Center resources; (4) communication, StrengthsFinder and personal interaction skills; and (5) Familiarity with the Food and Drug Administration (FDA) approval processes for clinical trials. These workshops/elements of training, which were universally praised, represent skills that are critical for success in interdisciplinary science and enhance success for all students because of the focus on and positive impact of networking. These five elements represent unique aspects of training designed for this training program based on UCI’s experience of successful clinical translation.

NIH Research Projects · FY 2026 · 2012-04

Project Summary The research outlined in this proposal seeks to further our understanding of the factors that govern C-H bond activation in cytochrome P450 catalysis. Over the past several years my group has made significant contributions to this area. Our results have impacted not only the way people think about P450 catalysis but also metal-oxo mediated C-H bond activation in general. We have led the way in the capture and characterization of critical intermediates in the P450 catalytic cycle and developed theories to describe how Nature biases enzymes for C-H bond activation. Still, much remains to be done. Our understanding of the factors that govern C-H bond activation in P450s remains incomplete. Importantly, results from our last funding period have shown that P450 can serve as a platform from which to attack some of the most important and fundamental questions in the field of C-H bond activation. There remains a debate in the field about the factors that govern reactivity in metal-oxo driven C-H bond activation. The debate centers on whether ground state thermodynamics, in the form of bond strengths, play the dominant role in determining reactivity or whether some other property of the system (e.g., spin density on the oxo ligand, metal-oxo basicity, access to low-lying excited states, or enhanced H-atom tunneling facilitated by strong electron donation from the axial ligand) can provide an intrinsic lowering of the activation barrier. The examination of this fundamental issue has been hindered not only by the difficulty of measuring these quantities for reactive high-valent species but also by the lack of a series of isoelectronic and isostructural compounds over which these quantities can be varied. We have shown that P450 can fill this void. During the previous funding period we expanded the range of P450s in which we can prepare compounds I and II for experimental studies. We now have access to an isoelectronic series of P450 intermediates with varying axial ligation. This and other innovations will allow us to measure the ground state thermodynamics of C-H bond cleavage, quantify the degree of H-atom tunneling and ferryl basicity, determine the amount of oxyl-radical character in compound I, examine the accessibility of low-lying excited states, and, importantly, track how these quantities (and the reactivity towards C-H bonds) change as a function of electron donation from the axial ligand. There is currently no other system, synthetic or biological, that allows for a similar set of measurements and discovery.

NIH Research Projects · FY 2026 · 2011-09

Project Summary/Abstract The long-term goal of this project is to develop novel approaches to treating meibomian gland dysfunction (MGD), a major cause of evaporative dry eye disease in the aging population. Meibomian glands are holocrine glands that are embedded in the tarsal plate of the both the upper and lower eyelid, and excrete lipid (meibum) onto the surface of the eye to form the lipid layer of the tear film to reduce aqueous tear evaporation. Dysfunction of the meibomian gland (MGD) identified as atrophy or altered meibum secretion is a common eyelid disorder having a widespread prevalence of 39-50% in the US population with the incidence increasing with age that is widely recognized as a major cause of evaporative dry eye disease (EDED). While patients with EDED and MGD comprise from 37% to 47% of the average Ophthalmologists and Optometrists practice, management of this disease is primarily palliative due to a lack of knowledge concerning the cellular and molecular events that cause MGD. To address this UNMET NEED we have evaluated the effects of conditionally knocking out (CKO) Pparg expression in the meibomian gland using tamoxifen induced Lrig1Cre/Ppargflox transgenic mice. Our studies show that following 10 weeks of Cre induction there is a male dominant (3/5 male to 0/35 female) ablation of acini with a shift from meibocyte differentiation to Krt6a+ ductal differentiation (See Preliminary Studies). These results support our 3D spheroid culture results concerning the plasticity of meibocyte/ductal epithelial differentiation as well as suggest that there are male/female differences in the regulation of meibocyte differentiation and renewal by stem/progenitor cells. Together, these findings suggest the following hypotheses: 1) That direct loss of Pparg signaling leads to an age and sex dependent loss of meibocyte differentiation, acinar atrophy and dry eye disease; 2) That Pparg signaling transcriptionally regulates meibocyte differentiation that when lost leads to up-regulation of ductal epithelial differentiation in progenitor meibocytes; 3) That loss of meibocyte progenitor cells recapitulates loss of Pparg leading to loss of meibocyte differentiation, acinar atrophy, and dry eye disease. To test these hypotheses, we propose the following Specific Aims: 1). Determine if loss of Pparg signaling in stem/progenitor meibocytes leads to acinar atrophy and dry eye by establishing the sex (male/female) and age (1, 3 and 6 months) effects of Ppargflox knockdown in Sox9-Cre and Lrig1-Cre mice; 2). Identify the Pparg specific transcriptional profile leading to meibocyte differentiation using scATACseq and ChIP-Seq by comparing wild type to Pparg CKO mice and identify differences related to meibocyte vs ductal differentiation as well as the effects of age and sex; and 3). Establish the effects of meibocyte stem/progenitor cell ablation by conditionally knocking out Sox9 and Lrig1 using Krt14-Cre, Sox9flox and Lrig1flox mice and evaluating the effects on acinar atrophy and dry.

NIH Research Projects · FY 2025 · 2011-07

Project Summary The voltage-gated proton channel Hv1 plays important roles in numerous biological processes, including pH homeostasis and the immune response. Its activity has been found to worsen brain damage after ischemic stroke, to exacerbate the effect of traumatic brain injury and spinal cord injury, and to increase the metastatic potential of different types of cancer. The development of small-molecule modulators of Hv1 activity could lead to new anti-inflammatory agents and anticancer drugs. In addition, Hv1 modulators can provide useful pharmacological tools for studying the function of the channel in health and disease. Hv1 belongs to the large family of voltage-gated ion channels (VGICs). The majority of these proteins consist of four voltage-sensing domains (VSDs) surrounding a central pore domain. While many types of drugs bind the pore domain of VGICs, the number of organic molecules known to bind VSDs is limited. The Hv1 channel is made of only two VSDs and does not contain a pore domain, providing a simplified model for studying how ligands interact with VSDs. We have previously discovered small molecules that inhibit Hv1 activity by binding within the intracellular vestibule of the channel VSD in the open state (class I.1 ligands). Using a rational design approach that combines experimental and computational methods, we identified related compounds that are able to bind the channel also in the closed state (class I.2 ligands). Some of the new ligands display inhibitory properties that are superior to those of class I.1 compounds and provide a promising scaffold for further development of high-affinity Hv1 antagonists. However, little is known about how effective class I.2 ligands are at inhibiting Hv1- regulated cellular processes, such as ROS production by NOX enzymes, or how specifically they target the Hv1 VSD versus VSDs of other VGICs. In aim 1 of this project, we will apply our rational design approach to develop I.2 ligands with improved potency and corresponding negative controls. We will also use electrophysiological methods to investigate potential effects of Hv1 ligands on other members of the VGIC family. In aim 2, we will utilize a variety of live cell imaging assays on wild type and Hv1 knockout cells to examine how I.2 ligands inhibit NOX-mediated ROS production in phagocytes and how they affect proliferation and migration of cancer cells in a Hv1-dependent manner. The Hv1 channel contains a VSD-VSD interface unique among VGICs. As a result, ligands that bind such interface are expected to be more specific channel modulators than ligands that bind other transmembrane regions. The structure of the Hv1 dimer has yet to be determined, and alternative dimer models have been proposed by different groups with different VSD-VSD interfaces. In aim 3, we will use molecular dynamics simulations combined with multichemistry cross-linking mass spectrometry to probe the different models and derive a consensus dimer interface.

NIH Research Projects · FY 2025 · 2010-09

PROJECT SUMMARY/ABSTRACT The circadian clock, an ancient, evolutionary conserved timing system required for optimal function of organs and organismal lifespan, is active in peripheral tissues, including the skin. Clocks in peripheral organs are coordinated by the central clock in the suprachiasmatic nucleus, but we also know that time-restricted feeding affects circadian clocks, gene expression, and homeostasis in peripheral tissues. Although new insights are emerging, especially from studies in metabolic organs like the liver, the interplay between feeding time, clocks, and tissue health in epithelia is unclear. In particular, we don't know how time-restricted feeding affects the regenerative function of epidermal stem cells and skin aging. In mice, the circadian clock coordinates progression of the cell cycle and DNA excision repair with intermediary metabolism, as reflected in the redox state of epidermal stem cells. Intriguingly, daytime-restricted feeding shifts the phase and decreases the amplitude of the skin circadian clock, and it shifts the expression of the metabolism-related transcriptome without altering the phase of the diurnal oscillations in DNA synthesis. Daytime-restricted feeding, then, disrupts the coordination between metabolism and cell cycle progression in epidermal stem cells. Whereas these cycles in epidermal stem cells are known to modulate the sensitivity to UVB-induced DNA damage, their role in homeostasis of epidermal stem cells remains otherwise unknown. Here, we will investigate the idea that the clock coordinates oscillations of metabolism-generated ROS levels with the cell cycle and the DNA repair machinery to maximize the health and function of epidermal stem cells. Specifically, we hypothesize that this regulation minimizes metabolism- generated ROS when most epidermal stem cells are undergoing DNA replication, the cell cycle stage most sensitive to oxidative DNA damage. This hypothesis predicts that daytime feeding-induced circadian misalignment in epidermal stem cells causes asynchrony between oxidative metabolism and the cell cycle, leading to increased ROS-induced DNA mutations, epidermal stem cell dysfunction, and skin aging. To test this hypothesis, we propose two aims. First, we will define the gene-regulatory mechanisms underlying time- restricted feeding modulation of the circadian clock and metabolism in epidermal stem cells. Second, we will determine how time-restricted feeding modulates epidermal stem cell function and affects the rate of age- associated DNA mutations in epidermal stem cells. The proposal is significant because it tests a new model of how the circadian clock coordinates the timing of intermediary metabolism and the cell cycle in epithelial stem cells to minimize the accumulation of somatic DNA mutations, and how time-restricted feeding can enforce or disrupt this coordination. The proposal is innovative because it pursues a new idea about the role of dietary intervention and the circadian clock in skin aging, and it uses state of the art approaches, including duplex DNA- sequencing, fluorescence lifetime imaging, and single cell RNA-sequencing--approaches not previously applied to skin aging.

NIH Research Projects · FY 2025 · 2010-09

SUMMARY/ABSTRACT Herpes simplex type virus-1 (HSV-1) infects over 3.72 billion people worldwide, including 200 million individuals in the United States. Following primary infection of the cornea, HSV-1 establishes latency in sensory neurons of the trigeminal ganglia (TG). Reactivation of HSV-1 from latently infected TG leads to shedding of the virus in tears causing recurrent ocular herpetic disease, a major cause of infectious blindness in the Western world. Currenly, an FDA-approved herpes simplex vaccine is unavailable. Our long-term goal is to develop an immunotherapeutic ocular herpes vaccine. While a role for CD8+ T cells (but not CD4+ T cells) in reducing HSV- 1 reactivations from latently infected TG is gaining wider acceptance, the small numbers of functional tissue- resident memory CD8+ TRM cells that are present in latently infected TG are not enough to prevent virus reactivation. We have made several significant findings, during the last funding period: (1) HSV-specific CD8+ T cells from “naturally protected” HLA-A*0201-positive asymptomatic individuals (who never develop recurrent ocular herpetic disease despite being infected) mainly targeted five HSV-1 epitopes; (2) Phenotypic and transcriptomicprofiling indicates that frequent HSV-specific CD8+ TRM cells, which expressed high levels of tissue-homing and tissue-residency receptors (i.e. CXCR3, IL-2R/IL-15R, CD69, and CD103), found in the TG of HSV-1 infected HLA-A*0201 transgenic rabbits (HLA Tg rabbits) are associated with decreased virus shedding; (3) Topical ocular delivery to latently infected HLA Tg rabbits of prototype neurotropic adeno- associated virus (AAV8) constructs, which express either the T cell attracting CXCL11 chemokine (CXCR3 ligand) or IL-2/IL-15 cytokines (IL-2Rb/IL-15Rb ligands), increased the frequency of TG-resident CD8+ TRM cells specific to the five immunodominant epitopes; (4) Increased numbers of exhausted TG-resident CD8+ TRM cells were associated with increased virus shedding in HLA Tg rabbits; and (5) Ex vivo blockade of T cells exhaustion pathways PD-1, LAG-3 and TIGIT, ex vivo, in rabbit TG explants significantly reduced virus reactivation. Building on the above published and preliminary results, the central hypothesis of this revised competitive renewal proposal is that a TG-targeted vaccine that boosts the number, function and longevity of anti-viral TG-resident CD8+TRM cells will reduce virus reactivation and shedding. Specific Aims: Aim 1: Test the hypothesis that a tissue-targeted Prime/Pull/Keep therapeutic vaccine (designated as PPK vaccine) that incorporates the five immunodominant HSV-1 CD8+ TRM cell epitopes (prime), CXCL11 (pull) and IL-2/IL-15 (keep) will boost the number and longevity of TG-resident CD8+ TRM cells and significantly decrease HSV-1 reactivation in latently infected HLA Tg rabbits. Aim 2: Test the hypothesis that tissue-targeted PPK vaccine combined with blockade of PD-1, LAG-3 and/or TIGIT immune checkpoints will increase the number of functional CD8+ TRM cells in the TG and produce even more robust protection in latently infected HLA Tg rabbits. This translational research is expected to pave the way towards developing a PPK vaccine to protect against recurrent ocular herpes in man.

NIH Research Projects · FY 2024 · 2010-07

Project Summary Progress towards understanding mechanisms of hearing, and the treatment of hearing deficits, requires an integrated effort from multiple disciplines. The Center for Hearing Research (CHR) at the University of California, Irvine (UCI) maintains an interdisciplinary training program that takes advantage of the breadth and depth of hearing research at UCI to train new scientists broadly across multiple disciplines and deeply in one. The 21 training faculty in CHR span six departments in five Schools (primary appointments in Biological Sciences, Medicine, Social Sciences and Education; joint appointments in Engineering) with research interests that cover a broad range of levels (genes, molecules, cells, systems and behavior) and experimental approaches (cell and molecular biology, neurophysiology, psychoacoustics, computation, human imaging, human learning, medical device engineering). Thus, CHR is ideally positioned to offer interdisciplinary training. We request support for three predoctoral students and two postdoctoral researchers including medical residents. The didactic core of the training program is a course in Auditory Neuroscience, which covers the auditory system from cells to psychoacoustics, cochlea to cortex, and basic to clinical sciences. Mandatory features of the training program that encourage interdisciplinary interactions are participation in all CHR activities (e.g., seminar series, journal club and annual conferences), presentations to scientifically diverse audiences, and regular meetings with basic and clinical scientists. The normal period of support for trainees is two years. Predoctoral trainees normally enter the program in their second year of graduate study and are required to take a course on grant writing and prepare an NRSA proposal. The program is managed by the Program Director and an Executive Committee, and will foster development of trainees’ intellectual, technical and professional skills needed to pursue successful careers in hearing research.

NIH Research Projects · FY 2025 · 2008-12

Regulation of cellular metabolic status is determined by the mTORC1 complex, which senses systemic nutrient levels and intracellular nutrients. We set out to define the signaling pathways controlling autophagy activation, and discovered that MAP4K3 phosphorylation of transcription factor EB (TFEB) dictates autophagy status in the cell, and documented that MAP4K3 autophagy regulation lies upstream of mTORC1 autophagy regulation. MAP4K3 activates mTORC1 when amino acids are plentiful, but the basis for this regulation is ill-defined. During the current funding cycle, we delineated the MAP4K3 – mTORC1 signaling pathway, linking MAP4K3 activation of mTORC1 to inhibition of AMP-activated protein kinase. Our findings reveal MAP4K3 inhibition of AMPK may occur via phosphorylation of Sirtuin-1 resulting in LKB1 inactivation. To understand the scope of MAP4K3 function, we completed an interactome and phosphoproteomics analysis, and implicated MAP4K3 in regulation of the GATOR1/2 complex, which controls mTORC1 localization to the lysosome, a prerequisite step for mTORC1 activation. We also determined that MAP4K3 can localize to the nucleus, and may participate in the DNA damage response. All these findings indicate that MAP4K3 is a central node for the regulation of cellular homeostasis, serving as a nexus for cross-talk between pathways of metabolism, cell stress, and cell survival. We have begun to examine the physiological relevance of MAP4K3 phosphoregulation by deriving lines of mice with phosphoresistant and phosphomimetic amino acid substitutions at the TFEB serine residue (S3) subject to MAP4K3 phosphorylation. As mTORC1 dysregulation is implicated in cancer and neurological disease, our results suggest that one appealing therapeutic strategy for diseases of altered mTOR signaling function would be to develop drugs to inhibit MAP4K3. To achieve this goal, we performed in silico screening for small molecules that interfere with MAP4K3 open pocket dimerization, and evaluated 13 compounds in a series of secondary and tertiary assays, identifying three promising hits. In this renewal project, we will define components of the MAP4K3 amino acid sensing dependent pathway of mTORC1 activation and delineate the molecular basis for MAP4K3 regulation of mTORC1 activation, focusing on Sirtuin- 1 phosphoregulation by MAP4K3 and the nature of MAP4K3 interaction with the GATOR1/2 complex. We propose to assess MAP4K3 regulation of autophagy and cell stress by carefully characterizing phenotypes and autophagy function in TFEB S3A and S3E mice, and determining if MAP4K3 is involved in modulating DNA damage responses. We will build on our MAP4K3 inhibitor translational drug discovery work by honing in on the most promising lead compounds through structure-activity relationship generation of a compound series, coupled with an independent in silico screen and kinase inhibitor potency testing followed by a critical path of validation assays, and we will test if our lead MAP4K3 inhibitor(s) can rescue disease phenotypes in a mouse model of tuberous sclerosis complex (TSC) and in frontotemporal dementia / tauopathy Tsc1 +/- mice .

NIH Research Projects · FY 2025 · 2007-12

Abstract The goal of this project is to improve the molecular diagnosis rate and better understand the molecular mechanisms of early onset cone inherited retinal degeneration (eocIRD) diseases. As human heavily relies on cone vision for the activities, degeneration of cone photoreceptors has significant impact on their ability to perform daily routine. Unfortunately, the current molecular diagnosis rate of eocIRD, such as cone rod dystrophy (COD) and cone rod dystrophy (CRD), is significantly lower that of rod degeneration IRDs such as retinitis pigmentosa. More than 50% COD and CRD remain unassigned even upon screening of all IRD associated genes, highlighting a significant gap in our knowledge of the disease. To overcome this challenge, we propose to systematically identify novel genes and mutant alleles that are missed by current screen process through a combination of short and long read whole genome sequencing and functional validation experiments. To achieve this goal, we have established a large collection of over 1,500 well-characterized, unrelated COD, CRD, and LCA patient families. Screens for mutations in known inherited retinal disease genes led to the identification of causal mutations in about 900 probands, leaving about 570 patient families unsolved. Patients from these families are likely to due to mutations missed by current technology, representing a well characterized, rich resource for identifying new mutations and disease associated genes. Whole exome sequencing has been performed for all unsolved probands, including 300 with whole genome sequencing. Building on this work, our Specific Aims are: Specific Aim 1. Characterize the novel eocIRD associated gene TLCD3B Specific Aim 2. Investigate the full spectrum of mutations in unsolved patients Specific Aim 3. Identify and perform functional studies of novel candidate disease genes Progress toward these goals is likely to lead to new insights into disease mechanisms through studies of novel eocIRD disease genes and lay the foundation for developing new diagnoses and treatment methods. Importantly, the protocols and software tools developed from these aims, particularly noncoding mutation identification, will be applicable to other human diseases as well.

NIH Research Projects · FY 2025 · 2003-07

Summary: We propose to renew our 20-year-old UCI Training Program in Epilepsy Research which supports a total of four predoctoral and postdoctoral trainees. Epilepsy is the third most common brain disorder, and one that affects the young preferentially, with a tremendous loss of human potential. Surprisingly, Epilepsy is relatively understudied guide and develop the next cohorts of diverse, outstanding trainees and provide them with state- of the art conceptual, technical, quantitative and scientific rigor approaches to the understanding and underfunded; thus, there is a major unmet need to encourage and train the new generation of the Epilepsy Investigator Workforce. Our program has strived and succeeded in shepherding a robust and diverse cadre of pre- and postdoctoral trainees who are now engaging in Epilepsy and Neuroscience Research and Education, the majority in Academia and Industry. We now propose to of Epilepsy and its co-morbidities, as well as professional skills for their career development. Our progress to date and the current proposal demonstrate that we have the experience, environment, resources and passion to accomplish these goals.

NIH Research Projects · FY 2025 · 2003-04

Abstract The pathogenic bacteria Pseudomonas aeruginosa is a major cause of blinding corneal infections in the USA and worldwide. This includes a recent outbreak of drug resistant P. aeruginosa keratitis in contaminated eyedrops that caused keratitis in 68 patients from 16 US states and resulted in 3 deaths and 8 enucleations. Our earlier studies supported by this grant demonstrated that P. aeruginosa virulence in infected corneas is due to expression of the Type III secretion system, which comprises a needle structure that injects exoenzymes directly into the host cell cytosol. Our recent data show that using either NLRP3-/- mice or the NLRP3 small molecule inhibitor MCC950, P. aeruginosa ExoS ADP ribosyl transferase (ADPRT) selectively licenses NLRP3 inflammasome usage in neutrophils (where macrophages use the NLRC4 inflammasome) for IL-1β processing and secretion, and importantly that NLRP3 is required for effective bacterial killing in infected corneas that can be blocked with the small molecule inhibitor MCC950. Proposed studies in Aim 1 will examine the early events in NLRP3 activation in neutrophils, including a role for NEK7 and HDAC6, which will also be examined during infection. Aim 2 will use an HEK293 based expression system to produce mg levels of NLRP3 that can used to identifying NLRP3 domains and specific arginine residues that are targeted by ExoS ADPRT. Aim 3 will examine cell death pathways in corneal epithelial cells and neutrophils infected with P. aeruginosa expressing the Type II secretion protein ToxA and determine if there is a role for NLRP1 pyroptotic cell death and in corneal infections. Results of these proposed studies will identify novel targets for immune intervention in this blinding infection.

NIH Research Projects · FY 2024 · 2002-09

PROJECT SUMMARY - PARENT GRANT (7/1/18 to 5/31/24) The 90+ Study was initiated January 1, 2003 as a population-based sample of oldest-old survivors of the Leisure World Cohort Study (LWCS, enrollment 1981-1984). With more than 1,800 participants, The 90+ Study is one of the largest and longest studies in the world of dementia, cognitive decline, disability, and frailty in the oldest-old. People over 90 are the fastest segment of the population and have the greatest public health impact as the risk of dementia is extraordinarily high in these individuals, reaching a staggering 40% per year in centenarians. However, many oldest-old maintain superior cognitive performance well into their tenth decade and beyond (cognitive resilience), often in the presence of neuropathological changes (cognitive resilience in the presence of pathology). Lifestyle, co-morbid conditions, genetics, and other factors have been implicated in this cognitive resilience but have not been well studied. In this application, we extend our studies to investigate cognitive resilience in these remarkable individuals. In Aim 1, we consider early (30 years earlier in the LWCS) and late (at age 90+) lifestyle and other factors in relation to cognitive resilience. Factors related to the maintenance of superior cognitive abilities in the presence of pathology are investigated in Aim 2. Taking advantage of our large cohort of 90+ year-olds in our imaging studies, we prospectively follow individuals without dementia to estimate incidence of dementia and rates of cognitive decline in relation to MRI and PET biomarkers in Aim 3. We analyze the role of low levels of multiple neuropathologic changes in the expression of dementia in Aim 4. Finally, with this application, we will recruit the last living LWCS participants eligible for our study, and complete ascertainment of all previously established outcomes in The 90+ Study (dementia, CIND, MCI, frailty, disability, and mortality). With our extensive database, unique large cohort of well- characterized individuals, extensive and multidisciplinary longitudinal follow-up, and innovative neuropathological and neuroimaging investigations, we are ideally positioned to do studies of dementia and resilience in the oldest-old. Identifying factors related to cognitive resilience and dementia, including modifiable lifestyle factors and imaging biomarkers, would contribute to our understanding of aging in health and disease and provide potential targets for interventions to promote successful aging.

NIH Research Projects · FY 2026 · 2002-05

Spinocerebellar ataxia type 7 (SCA7) is an inherited neurological disorder characterized by cerebellar and retinal degeneration. SCA7 is caused by CAG/polyglutamine (polyQ) repeat expansions in the ataxin-7 gene. Over 20 years ago, we linked SCA7 retinal degeneration to transcription dysregulation, and then discovered that ataxin-7 is a core component of the transcription co-activator complex STAGA, which possesses intrinsic histone acetyltransferase (GCN5) and histone deubiquitinase (USP22) activity. In the last funding cycle, we set out to determine the molecular basis of SCA7 cerebellar and retinal degeneration by examining the transcriptome and epigenome, and thereby implicated DNA damage, metabolic dysregulation, mitochondrial abnormalities, and calcium dyshomeostasis in SCA7 disease pathogenesis. To obtain a more complete understanding of the SCA7 disease process in cerebellum and retina, we are now pursing single-cell transcriptome and epigenome analysis in the highly representative SCA7 266Q knock-in mouse model. Employing a novel Purkinje cell-enriched single nucleus (sn)RNA-seq method, we discovered altered synaptic organization and excitatory-inhibitory balance in presymptomatic SCA7 mice. Bioinformatics analysis revealed dramatic dysregulation of normal aldolase-C / zebrin-II expression patterning and corresponding parasagittal striping in presymptomatic SCA7 mice, and based upon analysis of related SCA polyQ disease model mice, we established that altered cerebellar neurodevelopment is a shared disease feature across various polyQ cerebellar ataxias. Here we propose to define the epigenetic basis for transcriptional alterations in SCA7 retinal degeneration and cerebellar degeneration by performing snRNA-seq and snATAC-seq on retina and cerebellum samples from SCA7 mice, and classifying DEGs and DARs from different cell types to reveal genes and regulatory elements that underlie the molecular pathology. We will pursue Transcription Factor Binding Site analysis to define transcription factors (TFs) whose dysregulation may contribute to SCA7, and we will confirm candidate TF and epigenetic pathology by directed experimentation. We will modulate the expression or function of implicated TFs in SCA7 knock-in model mice to determine if such interventions can prevent or significantly ameliorate SCA7 neurodegeneration. This validation work will employ an in vivo epigenetic rescue strategy involving use of Cre-inducible dCas9-p300 (or -KRAB) mice, and we will determine if epigenetic rescue can ameliorate SCA7 retinal and cerebellar pathology, including disrupted zebrin-II patterning. Finally, we will test if interventions found to rescue SCA7 cerebellar disease can be applied to related polyQ SCAs.

NIH Research Projects · FY 2025 · 2001-09

Project Summary Our work funded by this RO1 during this last funding cycle has established that endogenous hydrogen sulfide (H2S) produced by selective upregulated H2S synthesizing enzyme cystathionine β-synthase (CBS) is a new uterine artery (UA) dilator system contributing to estrogen-induced and pregnancy-associated rises in uterine blood flow. This novel pathway has reshaped the view how uterine hemodynamics is regulated during normal pregnancy. More recently, we reported that H2S stimulates human UA relaxation via activating smooth muscle (SM) large conductance Ca2+-activated voltage-gated potassium (BKCa) channels; yet, how H2S mediates estrogen-induced UA dilation in normal and complicated pregnancies remains largely unknown. Formation of -SSH groups on reactive cysteine(s) in proteins, referred to as sulfhydration or persulfidation, has emerged as the main signaling route for H2S to exert its biological function. Sulfhydration converts free thiols (-SH) to persulfide (-SSH) resulting in increased reactivity of modified cysteines due to increased nucleophilicity of SSH compared with SH. In this competitive renewal RO1 application, we present new data showing that estrogen and pregnancy can significantly stimulate protein sulfhydration in human UA; and more interestingly, the elevated levels of total sulfhydrated proteins and sulfhydrated β1 and γ1 subunits of BKCa channels in human UA in normal pregnancy is significantly reduced in preeclampsia. Thus, we propose to test a novel hypothesis herein that augmented CBS/H2S sulfhydrates the increased β1 and γ1 BKCa (via estrogen receptor-dependent transcription) resulting in activation of SM BKCa to mediate estrogen-induced UA dilation in normal pregnancy and this mechanism is impaired in preeclampsia. This conjecture will be tested by two specific aims targeting on determining the estrogen-responsive BKCa channels and how sulfhydration results in activation of these BKCa channels pertaining to UA dilation in pregnancy and preeclampsia, with comprehensive biochemical, cellular, molecular, pharmacological, and physiological and electrophysiology approaches using in vitro primary cell culture models of UA smooth muscle cells, ex vivo studies of human main UA samples associated with different estrogens status from hysterectomy and myometrial UA samples from normal and preeclamptic pregnancies, and in vivo rat models to study the role of exogenous and endogenous estrogens in vivo. The proposed studies will establish a novel mechanism for BKCa channel activation via sulfhydrating its regulatory β1 and γ1 subunits to broadly impact on ion channel biology. These studies will comprehend specific mechanisms for activation of the estrogen-responsive SM BKCa channels pertaining to estrogen-induced UA dilation in pregnancy and preeclampsia. Data obtained will advance our understanding of estrogens and uterine blood flow biology, informing new pathways to assist the development of alternative strategies for combating preeclampsia. New data obtained will also shed lights on the understanding of the cardiovascular protective effects of estrogens. 1

NIH Research Projects · FY 2025 · 2001-04

Project Summary A functional skeletal system depends on the coordinated development of cartilages and bones during embryogenesis. However, little is known about the cellular and molecular mechanisms that control the polarized growth of cartilages, which determine endochondral bone size and shape. Unraveling the signals that direct mesenchymal cells to condense and align into pre-chondrogenic stacks is key to understanding early events that shape the organization and growth of the skeleton. Elucidating these processes will allow better diagnosis and treatments for skeletal malformations and birth defects. Moreover, molecules that control cartilage morphogenesis and differentiation may be of considerable clinical importance both for improvements in diagnosing and treating congenital birth defects as well as developing mesenchymal stem-cell based therapies for skeletal disorders. Evidence that planar cell polarity pathways are essential for cartilage cells to stack properly, suggests a previously unappreciated mechanism for patterning cartilage growth plates of long bones as well as growth zones in bones of the skull. Dramatic results from many laboratories now demonstrate that Dlx transcription factors regulate planar cell polarity signaling, well known for its critical roles in long bone growth plates, as well as craniofacial cartilage polarity in zebrafish. Embryos deficient in Dlx5 and Dlx6 show defects in cartilage stacking, like mutants in Wnt5b and Ror2. Moreover, comparisons of cartilage growth zones in African cichlid fishes that have evolved dramatically different craniofacial bone shapes, reveal that growth zone size differences during larval development correlate with these species-specific shapes. Aim 1 will build upon previously funded work to address the hypothesis that Dlx5/6 directly regulate pharyngeal arch patterning and growth zone formation via planar cell polarity. Cartilage phenotypes will be evaluated in embryos and larvae in which Dlx5/6 have been genetically deleted and identify the polarity pathways regulated by Dlx5/6 as well as the signaling and responding cells. Aim 2 will address the functions of Wnt5- as well as Fat3-mediated planar polarity in early arch morphogenesis as well as later growth zones, including both transcriptional and non- transcriptional modes of propagation such as cytosplasmic extensions called cytonemes. New transgenic tools will allow tracking of polarity and visualization of cytonemes. Finally, Aim 3 will continue “evo-devo” projects focused on discovering new genes involved in cartilage polarity and growth zones using quantitative trait locus mapping in cichlids. Together, these studies will lead to mechanistic insights into the relatively unexplored functions of cellular polarity in the vertebrate skeleton. This work will lead to insights into the causes of human skeletal disorders of Dlx5/6, such as Split Hand Foot syndrome, as well as polarity disorders such as Robinow and Van Maldergem syndromes.

NIH Research Projects · FY 2025 · 2000-09

Project Summary Malaria remains the most devastating vector-borne disease in Africa. The primary interventions, insecticide-treated nets and indoor residual spraying, target indoor-biting mosquitoes. However, the effectiveness of these tools has been significantly hindered by the spread of insecticide resistance and the increasing tendency of malaria vectors to bite outdoors, slowing malaria control progress across Africa. Additionally, the recent invasion of Anopheles stephensi, a major malaria vector in South Asian urban environments, presents a new and significant threat to malaria control efforts in Africa. This invasion, along with rapid urbanization on the continent, has created additional challenges for malaria control in Africa. To address these challenges, it is essential to understand the biology of An. stephensi in its newly-invaded African habitats, its impact on malaria epidemiology, and to develop new vector control methods suited to urban environments. Advances in molecular biology, genomics, bioinformatics, and ecological modeling offer exciting opportunities to track and contain the spread of An. stephensi and develop new control methods in rapidly growing urban areas. Unfortunately, many scientists from malaria-endemic countries have not fully leveraged these technologies in their research. In previous funding cycles, our D43 research training program has achieved remarkable success in training African scientists in vector biology and malaria research. However, significant gaps remain, particularly in the research capacity for invasive vector species in Ethiopia and other African countries. Accordingly, the overall training objectives are to advance the career development of promising young scientists from An. stephensi-infested African countries, strengthen institutional research capacity in invasive disease vectors and promote sustainability among malaria researchers in Ethiopia. We plan to achieve this by training ten PhD students, four postdoctoral fellows and 20 junior scientists from these regions through mentored research in vector biology and malaria epidemiology, and through workshops and short and intensive training courses on grant writing and scientific leadership development. In addition to mentored research, the program will offer a core curriculum focusing on biostatistics, genomics, bioinformatics, modeling, scientific writing, and responsible conduct of research. The infrastructure and capacity at our international training sites in Ethiopia and at the University of California, Irvine, are ideal for this training. This program will significantly enhance institutional research capacity in invasive disease vectors and malaria in Ethiopia and aid the career development of African scientists by bridging laboratory and field research in vector biology and malaria epidemiology, enabling researchers with innovative technologies, fostering Africa-wide and international collaborations, and supporting them to become independent scientists.

NIH Research Projects · FY 2025 · 1998-08

PROJECT SUMMARY/ABSTRACT Although the epidermis is composed of morphologically distinct layers, single-cell RNA-seq experiments indicate that epidermal differentiation does not progress in discrete steps at a transcript level but is instead continuous and gradual. Consistent with this notion, we identified a large population of basal-to-spinous transition cells that reside in the basal and first spinous cell layers. These transition cells are found throughout the lifetime of the mouse and in the adult human palmoplantar and non-palmoplantar epidermis. Our hypothesis is that the presence of these transition cells does not reflect an ineffectiveness in switching from basal to spinous gene- regulatory programs. On the contrary, we propose that basal-to-spinous cells are actively maintained through a specific gene-regulatory program. We will use single-cell and spatial transcriptomics approaches to study the cellular composition, gene expression, and predicted function and cell-cell signaling in the P0 epidermis from mice knocked out for transcription factors whose expression peaks in transition cells. We will use single-nuclear ATAC-seq with single-nuclear RNA-seq to map changes in open chromatin across cell states, including in basal- to-spinous transition states. We will also define transcription factor binding and histone modifications in sorted epidermal cells from distinct differentiation stages. We will then use computational modeling to define the underlying gene regulatory networks and to inform further functional testing on the role of signaling pathways and transcription factors in maintaining the basal-to-spinous transition cell state. This work is significant and innovative because it addresses a recently recognized cell state in epidermal differentiation, basal-to-spinous transition cells, which have not been investigated.

NIH Research Projects · FY 2026 · 1997-09

OVERALL: DIRECTOR'S OVERVIEW & SIX ESSENTIAL CHARACTERISTICS PROJECT SUMMARY/ABSTRACT The Chao Family Comprehensive Cancer Center (CFCCC) of the University of California, Irvine (UCI) is the only NCI-designated cancer center based in Orange County, the 6th most populous county in the U.S. that serves as our Catchment Area and is home to ~1% of the nation's people. Now entering its 27th year, the CFCCC will enact its new Strategic Plan to enhance its role as a vital resource for the people of Orange County and surrounding areas in southern California to generate and disseminate new knowledge about the causes, prevention, and treatment of cancer, to train of the next generation of cancer providers and caregivers, and to alleviate the overall burden of cancer on our residents. The CFCCC has 195 members drawn from 32 academic departments across nine Schools at UCI, including the Schools of Medicine, Nursing, Pharmacy & Pharmaceutical Sciences, Population & Public Health, Biological Sciences, Physical Sciences, Information & Computer Science, Engineering, and Business. Our parent institution, UCI, has been consistently ranked in the top 10 of public universities in the U.S., while our CFCCC members are supported by $29.2 million in total annual extramural peer-reviewed cancer research funding, including $9.3 million from the NCI. Our members published 1,792 cancer-focused, peer-reviewed publications from 2015-2020, of which 73% are collaborative and 20% are in high-impact journals. CFCCC research is organized into three Research Programs that provide an interactive and collaborative infrastructure for cancer discovery, clinical investigation including early phase and investigator-initiated trials, and population-based cancer research. These include two Programs spanning basic, translational, and clinical cancer research, Biotechnology, Imaging & Drug Development (BIDD) and Systems, Pathways & Targets (SPT), as well as Cancer Control (CC), our population science Program. The Programs are further linked by seven CFCCC Disease-Oriented Teams that bring together basic, translational, clinical and population investigators to facilitate the movement of CFCCC discoveries through the pipeline into the clinical arena. CFCCC research is supported by seven Shared Resources that provide our members access to specialized services, technology and instrumentation, and expert consultation and collaboration. These Shared Resources include the Transgenic Mouse Facility, Optical Biology Core, Genomic High-Throughput Facility, In Vivo Functional Onco-Imaging, Experimental Tissue Resource, Biostatistics Shared Resource, and the Biobehavioral Shared Resource. Clinical research is supported by the Stern Center for Cancer Clinical Trials & Research, the CFCCC clinical trials office. With this competing renewal application, the CFCCC requests CCSG funding for the next 5 years to support our Administration, Developmental Funds, Program and Senior Leadership, Community Outreach & Engagement, Cancer Research Training & Educational Coordination, Shared Resources, Clinical Protocol & Data Management, and Protocol Review & Monitoring, to allow the CFCCC to continue its mission.

NIH Research Projects · FY 2025 · 1994-04

The broad purpose of the research in this proposal is to understand how microenvironrnents (secondary coordination spheres) about metal ions control function. A bio-inspired synthetic approach is utilized that incorporates principles of molecular architecture found in the active sites of metalloproteins into synthetic systems. Multidentate ligands will be developed that create rigid organic structures around metal ions and place hydrogen bond donors or acceptors proximal to the metal centers, forming specific microenvironments. One distinguishing attribute of these systems is the ability to make site-specific modifications to the structure in order to evaluate correlations between the microenvironment and reactivity. A focus of this research is the examination of transient intermediates that are formed from the activation of dioxygen and the oxidation of water - processes that are directly linked to the maintenance of human health and aging. Long-term goals include developing structure-function relationships in metal-assisted oxidative catalysis. Metalloproteins perform functions not yet achieved in synthetic systems. Our hypothesis is that the lack of control of the secondary coordination sphere in synthetic compounds is a major obstacle in establishing the desired functions. Results from structural biology show that hydrogen bonds within the secondary coordination spheres of metalloproteins are instrumental in regulating function. Therefore, the function and dysfunction of health-related metalloproteins can be understood in the context of changes in their microenvironrnents. However, it is still unclear, even in biomolecules, how non-covalent interactions influence metal-mediated processes. Investigations into these effects require fundamental reactivity and mechanistic studies in which the contributions of single components can be analyzed individually. We have developed synthetic hydrogen bonding systems in which the molecular components that define the structure around the metal ion are specifically controlled; in turn, this permits the formation of systems whose activity can be tailored to a particular function. This ability to regulate the microenvironment allows for systematic studies into structure- function relationships that lead to fundamental understanding of chemical processes. Ultimately, this research will provide insights into the properties of biological catalysts and lead to new classes of synthetic catalysts that exhibit the exquisite control over reactivity that is characteristic of metalloenzymes.