In vivo 2-photon imaging of retinal biochemistry before and after retinal organoid transplantation

视网膜类器官移植前后视网膜生物化学的体内2光子成像

基本信息

批准号：
10643273
负责人：
Andrew W Browne
金额：
$ 19.43万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10643273
关键词：
Adult Age Age related macular degeneration Animal Model Award Biochemical Biochemistry Biological Markers Biological Sciences Biology Biophotonics Bioreactors Blindness California Capital Cell Death Cell physiology Cells Cellular biology Cessation of life Chemistry Clinical Clinical Assessment Tool Collaborations Data Development Development Plans Disease Elderly Electrophysiology (science)Elements Engineering Ensure Equipment Exhibits Eye Eye diseases Fluorescence Fluorescence Microscopy Fostering Functional Imaging Functional disorder Future Gene Mutation Genetic Genetic Diseases Goals High Pressure Liquid Chromatography Histology Human IACUC Image Imaging Device Imaging Techniques Immunohistochemistry In Vitro Inflammation Investigation Knowledge Label Laboratories Laboratory Research Light Link Mentors Metabolic Metabolism Methods Microfluidics Microscope Modeling Molecular Molecular Analysis Molecular Profiling Monitor Multimodal Imaging Mus Mutation Operative Surgical Procedures Optics Organoids Phase Photoreceptors Physicians Population Procedures Protocols documentation Quality Control Rattus Recovery Rehabilitation therapy Replacement Therapy Reporter Reporter Genes Research Research Activity Research Personnel Resources Retina Retinal Degeneration Retinal Diseases Retinoids Scientist Source Structure of retinal pigment epithelium System Techniques Testing Therapeutic Time Tissue Transplantation Tissues Training Training Programs Training Support Transgenic Organisms Translating Translational Research Translations Transplantation Universities Vision Visual Visualization Work biomarker identification blind career career development cell replacement therapy cell type doctoral student engineered stem cells experience imaging biomarker imaging modality improved in vivo insight invention metabolic imaging mouse model next generation non-invasive imaging novel novel therapeutics optical imaging organoid transplantation pre-clinical research preclinical study preclinical trial predictive marker professor replacement tissue restoration retina transplantation retinal imaging skills stem cells success superior colliculus Corpora quadrigemina therapeutic candidate tissue culture tool transcriptomics transplantation therapy treatment response two-photon

项目摘要

Project Summary Candidate and Career Development Plan: Dr. Browne is an assistant professor practicing vitreoretinal surgery and functioning as an engineer and physician-scientist at the University of California, Irvine. Dr. Browne established his engineering and basic biological sciences skills as a PhD student. He has balanced clinical duties with his laboratory research activities and. His long-term career goals are to engineer functional imaging tools that will advance the understanding of early and advanced eye diseases and facilitate the therapies needed to treat humans. His training thus far has been using in vitro imaging alone. To achieve his career goals, he is requesting support for training to develop functional imaging tools in vivo and the molecular tools to correctly interpret Two-photon imaging (2PI) observations from animal models. This K08 award will enable Dr. Browne to develop his scientific and professional skills in advanced imaging, fluorescence microscopy, and retinal cellular biology applied in vivo. Dr. Browne and his co-mentors, Drs. Palczewski, Seiler and Kuppermann, have developed a hands-on strategy to fulfill the training requirements through relevant course work, didactics, laboratory techniques, and collaborations at UC Irvine. The training program will prepare Dr. Browne to submit R01-level proposals to independently investigate and optimize functional retinal imaging as a tool for therapeutic discovery. Research Plan: Many blinding retinal conditions, like age-related macular degeneration, are the consequence of biochemical dysfunction and the secondary effects of inflammation and cell death. Conventional clinical tools provide valuable structural information about the retina but are unable to visualize retinal function. Non-linear optical imaging can now reveal subcellular biochemistry in vitro and has emerged as a reliable tool to study retinal disease. 2PI in mouse models demonstrates subcellular changes in energy and light cycle metabolism. Cell replacement therapy has emerged as a therapeutic candidate for advanced vision loss, and preclinical trials in rats have demonstrated visual function restoration. To understand metabolic function in retinal organoids (RtOg) before and after transplantation into blind rats, we will employ functional 2PI in vitro and in vivo as outlined in the following specific aims. 1) Investigate RtOg maturation with 2PI and correlate functional imaging data with cell-specific reporters in vitro and molecular signatures post vivo, 2) Identify functional imaging biomarkers for in vivo metabolism using 2PI of normal rats and rat with retinal degeneration, 3) Study in vivo functional 2PI of transplanted RtOgs in rats to identify alterations in functional imaging biomarkers and correlate imaging findings with visual function testing. Completion of these aims will yield time-resolved metabolic detail for specific cell populations in developing RtOgs, avail novel in vivo information about retinal biochemistry in healthy retina, diseased retina, and diseased retinas treated with retinal sheet transplantation. These discoveries will guide future translation of both functional imaging and tissue replacement therapy in humans.

项目摘要候选人和职业发展计划：布朗博士是助理教授从事玻璃体视网膜手术并在加利福尼亚大学尔湾分校担任工程师和医师科学家。布朗博士在博士生中建立了他的工程和基本生物科学技能。他已经平衡了临床职责随着他的实验室研究活动和。他的长期职业目标是设计功能成像工具这将提高对早期和晚期眼部疾病的理解，并促进所需的疗法对待人类。到目前为止，他的培训一直在使用体外成像。为了实现他的职业目标，他是要求支持培训以在体内开发功能成像工具和分子工具正确解释来自动物模型的两光子成像（2PI）观察结果。这个K08奖将使布朗博士能够发展他在高级成像，荧光显微镜和视网膜细胞方面的科学和专业技能生物学在体内应用。布朗博士及其联合给出者，博士。 Palczewski，Seiler和Kuppermann，制定了动手策略来通过相关课程工作，教学， UC Irvine的实验室技术和合作。培训计划将准备布朗博士提交 R01级的建议，以独立研究和优化功能性视网膜成像作为治疗的工具发现。研究计划：许多令人眼花vent乱的视网膜状况，例如与年龄相关的黄斑变性，是结果生化功能障碍以及炎症和细胞死亡的次要作用。常规临床工具提供有关视网膜的有价值的结构信息，但无法可视化视网膜功能。非线性光学成像现在可以在体外揭示亚细胞生物化学，并已成为可靠的研究工具视网膜疾病。小鼠模型中的2PI显示了能量和光周期代谢的亚细胞变化。细胞置换疗法已成为高级视力丧失的治疗候选者和临床前试验在大鼠中显示了视觉功能恢复。了解视网膜器官中的代谢功能（RTOG）移植到盲大鼠之前和之后，我们将在体外和体内使用功能2PI，如概述在以下特定目标中。 1）使用2PI研究RTOG成熟，并将功能成像数据与体内体外和分子特征的细胞特异性记者，2）识别功能成像生物标志物使用正常大鼠的2PI和带有视网膜变性的大鼠的体内代谢，3）研究体内功能2pi的研究大鼠移植的RTOGS以识别功能成像生物标志物的变化并将成像发现相关联进行视觉功能测试。这些目标的完成将产生特定细胞的时间分辨的代谢细节开发RTOG的种群，可用新颖的体内信息，以了解健康视网膜中的视网膜生物化学，病态的视网膜和视网膜片移植治疗的视网膜病变。这些发现将指导人类功能成像和组织替代疗法的未来翻译。