University Of California-Irvine

NIH Research Projects · FY 2025 · 1992-08

Vision is initiated when retinal photoreceptors respond to light via isomerization of the opsin-bound visual chromophore 11-cis-retinal (11cRAL), releasing all-trans-retinal and triggering signal-transduction that transmits visual information to the brain. Sustained vision requires continuous renewal of the chromophore, originally thought to occur exclusively through the canonical visual (retinoid) cycle. Recent research advances have identified alternative pathways for production of 11cRAL in the retinal pigment epithelium (RPE) and Müller cells and developed gene-therapy approaches to restore 11cRAL synthesis in the setting of disease. These new findings have raised key questions regarding the flow of 11cRAL and its metabolites within and between the RPE, photoreceptors, and Müller glia, as well as the long-term efficacy of gene therapy to correct visual cycle deficits. We will address these questions by using novel mouse models that enable study of subcellular distribution of retinoids with unprecedented resolution and chemical specificity, and by applying newly developed genome-editing technologies to compare the efficacy and longevity of gene alteration/correction versus currently available gene-augmentation techniques to rescue retinal-disease models related to retinoid metabolism. Aim 1: Elucidate the trafficking of 11-cis-retinoids within the neural retina. We developed a novel retinoid- trapping method to isolate retinol isomers within specific cells and subcellular compartments, enabling robust detection of these species by HPLC and in vivo two-photon microscopy, even when they are present as transient intermediates under physiological conditions. This approach will enable definitive resolution of hypothesized retinoid trafficking pathways that have, to date, not been possible to probe experimentally. Aim 2: Determine the efficacy of genome-editing versus gene-supplementation therapy in rd12 mice. Gene therapy is gaining prominence for treating deficiencies in visual-cycle enzymes. However, gene- augmentation therapy has several drawbacks, including limited durability and persistence of the dysfunctional gene. Genome editing can mitigate these shortcomings. We will compare the efficiency and durability of both treatments. Mice expressing the mutant Rpe65 gene will be treated at different ages with Rpe65-AAV vectors, or with virus-like particles (VLPs) delivering the genome editor; and therapeutic durability will be tested over time. Aim 3: Develop genome-editing therapy for Rpe65-associated autosomal dominant retinitis pigmentosa (adRP). We made an adRP mouse model that expresses the c.1430A>G (D477G) mutant of Rpe65. This mutation serendipitously introduces a protospacer adjacent motif (PAM), enabling us to use CRISPR-Cas9 to selectively inactivate the mutant gene and restore function. Alternatively, we will use prime editors to correct the RPE65 deficiency with a long substitution extending from the PAM sequence. Editors and guide RNA will be delivered using VLPs and lipid nanoparticles carrying Cas9-ribonucleoprotein cargo. We will optimize the editing efficiency and evaluate treatment efficacy in restoring normal function in this adRP disease model.

NIH Research Projects · FY 2024 · 1988-09

This proposal is a competing renewal application for continuing support of a T32 training grant entitled “Virus- host interactions: a multi-scale training program.” The program, formerly called Molecular Biology of Eukaryotic Viruses, was established at the University of California, Irvine (UCI) in 1988. During its 30-year history, the goal of this program has been to broadly train Ph.D. graduate students in the fundamental aspects of molecular and cellular virology as they relate to virus-host interactions, viral pathogenesis, regulation of viral gene expression, virus structure/proteomics, and applications of virus/phage technologies. During the current review period (9/14-8/19), eight predoctoral students were supported by this training program. For the upcoming budget period, three pre-doctoral trainee positions are requested, consistent with previous renewals of this program. Due to evolving faculty research expertise and the emergence of new virus/phage- related disciplines, the scope of proposed training areas will be broadened to also include interdisciplinary research and training programs in host responses to viral infections, bacteriophages in the human microbiome and the environment, and mathematical modeling of virus dynamics. There are 13 faculty mentors for this program, representing eight academic departments at UCI. Nearly all faculty members from these departments participate in the Cellular and Molecular Biosciences graduate program (CMB) at UCI, which oversees recruitment, admission, and first-year training of predoctoral students. The majority of the T32 training grant trainees are part of the Immunology and Microbiology focus area in the CMB program. Three of the training preceptors are affiliated with the Department of Ecology & Evolutionary Biology, and research in these labs will focus on (i) viruses/bacteriophages in the environment, (ii) viruses associated with the human microbiome, and (iii) in vivo virus dynamics. All trainees in the proposed training program will take formal course work that includes an integrated set of core courses in molecular virology/pathogenesis, phage-host interactions, mathematical modeling of virus spread, and a Virology Journal Club as well as electives and seminar courses in their areas of research specialization. The virology training faculty have major research strengths in virus-host interactions including the immune response to viral infections, regulation of viral gene expression, structural virology and viral proteomics, viral pathogenesis, control of emerging virus infections, viruses/bacteriophages in the human and environmental microbiome, modeling of virus populations, molecular evolution of viruses, virus gene therapy, and drug delivery via viral vectors. Our research programs cover a wide range of DNA and RNA viruses, including herpes simplex virus, varicella zoster virus, vaccinia virus, adeno-associated virus, bacteriophages, retroviruses (HIV), filoviruses, arenaviruses, flaviviruses, and picornaviruses.

NIH Research Projects · FY 2025 · 1982-09

PROJECT SUMMARY/ABSTRACT At present 6.7 million people in the US are living with Alzheimer’s disease and countless others are impacted by age-related cognitive decline. The cost of care for these is skyrocketing. We urgently need to discover new prevention and treatment strategies. To do this, we need to train the next generation of scientists in modern techniques and in how to integrate findings across multiple domains and research approaches. UCI has a long and distinguished history in the study of brain plasticity and learning and memory at multiple levels and in integrating across levels and approaches. In addition, UCI has long and growing expertise in neuroinflammation, stem cell biology and translational science, all of which are included in this training program. However, new areas of strength have been developing and collaborations formed with enhanced animal and human imaging, sleep researchers, cognitive sciences and clinical trials that is vital to real progress in generating the capabilities to identify, treat and prevent/delay pathological decline in cognitive function. We seek to provide pre- and postdoctoral trainees with training in concepts such as the integrative roles of genetics and environment in brain aging and as a function of lifespan. Although a significant fraction of research involves animal studies, we emphasize a broadened consideration of "bridging the gap" between animal and human studies. Overall, our training program has six primary features and strengths: 1) A team of innovative and scholarly preceptors who have a strong record of accomplishment for training young scholars; 2) An excellent collaborative environment; 3) A core set of courses on tailored to brain aging and dementia research; 4) A set of seminars and symposia and training on brain pathology through Clinical-pathological case presentations 5) Specific training in oral and written communication skills; and 6) Individual guidance and mentoring to help trainees reach their career goals. Overall, our Training program in Neurobiology of Aging and Alzheimer’s disease is designed to develop a uniquely trained cadre of investigators who over the years will develop successful careers bench to clinic translation of research to improve the lives of the elderly and ensure sustainably healthcare for the nation.

NIH Research Projects · FY 2025 · 1980-07

Support is requested to continue a successful cancer research training program at the University of California, Irvine (UCI). This newly renamed Training Program for Interdisciplinary Cancer Research (IDCR) builds on a foundation of more than 40 years of experience developing PhD students and postdoctoral fellows to become scientific leaders. As the nature of cancer research and treatment have evolved, so has the focus of our training program. We emphasize interdisciplinary approaches to advance knowledge in cancer biology and treatment, offering trainees a highly collaborative environment that includes faculty mentors from five different Schools at UCI. Coursework emphasizes critical analysis of research literature in cancer and related disciplines, the most current research methods, and a focus on translational science such as the development of therapeutics and diagnostics. The scientific foundation we provide together with professional development activities offered will produce a cohort of well-trained experts armed to successfully attack the cancer problem from vantage points in both academia and industry. The Program benefits from outstanding institutional support from the UCI Chao Family Comprehensive Cancer Center (a NCI-designated comprehensive cancer center), the UCI Cancer Research Institute, Graduate Division, and other campus offices. The Program provides research opportunities across the cancer continuum from etiology to therapeutics, encompassing faculty from the Schools of Biological Sciences, Medicine, Pharmacy and Pharmaceutical Sciences, Physical Sciences, and Engineering. There are thirty training faculty, each with a cancer-focused research program and extramural funding. We request support for four postdoctoral and four predoctoral trainees; one additional predoctoral position will be supported by our Graduate Division. The Program is led two co-Directors, both senior faculty with outstanding records of research productivity and training, and a history of collaboration. In preparation for this renewal application, we have carried out a rigorous self-evaluation with input from trainees, faculty, and externaladvisors. This process has led us to implement a revised program that meets the specific needs of current UCI trainees. A continuing element of the Program will be a rigorous and well-defined set of courses (“Cancer Biology parts A and B”, "Clinical Cancer for Basic Scientists") that build knowledge about basic and clinical/translational cancer research. Other strong components that will continue are the biannual program retreat, the annual symposium in basic cancer research, and access to exceptional professional development opportunities (established under a NIH-BEST award and continuing with campus support). A fundamental change will be to redesign our journal clubs and research-in-progress talks to maximize interaction with diverse training faculty and expand active learning of interdisciplinary cancer research approaches. We will also implement several changes in program administration to increase faculty participation and to optimize mentoring, evaluation, and recruitment.

Other NSERC · FY 2024

visual perception, visual confidence, metacognition, visual psychophysics, perceptual decision-making, visual performance fields, signal detection theory, accumulation of evidence models, computational modeling, visual information processing