The role of miRNAs in retinal progenitor cell cycle regulation and competence

miRNA 在视网膜祖细胞周期调节和能力中的作用

• 批准号: 9759692
• 负责人: Corinne L. Fairchild
• 金额: $ 6.77万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759692
• 关键词: Address; Adult; Age related macular degeneration; Area; Blindness; Categories; Cell Cycle; Cell Cycle Kinetics; Cell Cycle Regulation; Cells; Cessation of life; Chemicals; Chick Embryo; Chickens; Competence; Cone; Core Facility; Data; Degenerative Disorder; Development; Educational process of instructing; Electroporation; Embryo; Embryonic Development; Environment; Faculty; Flow Cytometry; Gene Expression; Goals; Grant; Human; Image; In Vitro; Legal Blindness; Length; Lesion; Mediating; Mentors; Mentorship; MicroRNAs; Molecular; Nervous system structure; Neurons; Nucleic Acids; Outcome; Pathway interactions; Patients; Play; Positioning Attribute; Primates; Process; Production; Protocols documentation; Replacement Therapy; Reporter; Research; Research Personnel; Research Training; Retina; Retinal; Retinal Cone; Role; Science; Stem cells; Structure; Technical Expertise; Techniques; Testing; Time; Vision; Work; Writing; base; career development; cell type; collaborative environment; experience; experimental study; fovea centralis; human disease; improved; in vivo; insight; knockout animal; macula; nerve stem cell; neural circuit; neuroregulation; notch protein; outreach; progenitor; retinal imaging; retinal progenitor cell; retinogenesis; skill acquisition; skills; success; transcriptome sequencing

PROJECT SUMMARY Proper visual function relies on all the cell types in the retina being generated in the correct proportion and at the correct developmental time. However, our understanding of how neural progenitors determine which cell fates to produce during retinogenesis in the embryo has remained ambiguous. We know that retinal cells are generated from a common pool of retinal progenitors that shift their competence over time to give rise to seven retinal cell types in a stereotypic sequence. But, we do not know the mechanisms that control competence transitions, and our understanding of retinal cell fate determination is incomplete. Recent studies have shown that microRNAs (miRNAs), short, single stranded nucleic acid molecules that negatively regulate gene expression, are required for retinal progenitor cells to transition from producing early- to late-born retinal cell types; but, we do not know their mechanism of action. My preliminary data suggests that miRNAs regulate cell cycle kinetics in retinal progenitors, and has led to my hypothesis that miRNAs mediate progenitor competence by controlling cell cycle length. In this proposal, I will test my hypothesis via the following Aims: Aim 1: To determine how late-progenitor miRNAs regulate cell cycle dynamics in retinal progenitors. Aim 2: To determine how retinal progenitor cell cycle length impacts cell fate. Aim 3: Determine if miRNA-directed changes in cell cycle dynamics mediate the development of the chicken high acuity area. I will use a combination of in vivo and in vitro techniques; including, flow cytometry, live imaging of retina explants, RNA-sequencing and chick embryo electroporation to answer my research questions. This will greatly expand my technical skillset, which will help me become a more competitive candidate for a faculty position. In addition to expanding my technical skills, in my research training plan I address the soft skills required for my success as an academic researcher by assembling a group of experienced mentors who will provide mentorship in the following categories: Career development, teaching, grant writing and science outreach. UC Davis provides a very collaborative environment and access to top-notch core facilities; thus, it is an ideal environment for me to perform my experiments and enhance my career development skills. Successful completion of the project outlined here will provide crucial insight into the mechanisms that control neural progenitor cell fate decisions. This information is crucial to establish efficient protocols for generating donor cells from stem cells, which can subsequently be used for cell replacement therapies to treat degenerative diseases like age-related macular degeneration.

项目总结 正确的视觉功能依赖于视网膜中的所有细胞类型都以正确的比例和 正确的发育时间。然而，我们对神经前体如何决定哪些细胞的理解 胚胎在视网膜发生过程中产生的命运仍然不明确。我们知道视网膜细胞 由一个共同的视网膜祖细胞池产生，随着时间的推移，这些祖细胞的能力会发生变化，产生七个 视网膜细胞类型按常规顺序排列。但是，我们不知道控制能力的机制 转变，我们对视网膜细胞命运决定的理解是不完整的。最近的研究表明 MicroRNAs(MiRNAs)，一种负向调节基因的短单链核酸分子 表达，是视网膜前体细胞从产生早期视网膜细胞过渡到晚期出生视网膜细胞所必需的 但是，我们不知道它们的作用机制。我的初步数据表明miRNAs调节细胞 视网膜祖细胞的循环动力学，并导致了我的假设，即miRNAs介导祖细胞的能力 通过控制细胞周期长度。在这项提议中，我将通过以下目标来检验我的假设： 目的1：确定晚期祖细胞miRNAs如何调节视网膜祖细胞的细胞周期动力学。 目的2：研究视网膜祖细胞周期长度对细胞命运的影响。 目的3：确定miRNA诱导的细胞周期动力学变化是否参与鸡的发育 高敏感区。 我将使用体内和体外技术的结合；包括流式细胞术，视网膜的实时成像 外植体、RNA测序和鸡胚电穿孔来回答我的研究问题。这将是 大大扩展我的技术技能，这将帮助我成为一个更具竞争力的教职员工候选人 位置。除了扩展我的技术技能外，在我的研究培训计划中，我还讨论了软技能 作为一名学术研究人员，我需要召集一群经验丰富的导师，他们将 在以下类别中提供指导：职业发展、教学、赠款撰写和科学 外展。加州大学戴维斯分校提供了高度协作的环境和访问一流核心设施的途径；因此， 一个理想的环境，让我进行我的实验，提高我的职业发展技能。 这里概述的项目的成功完成将提供对控制 神经前体细胞命运的决定。此信息对于建立有效的协议以生成 干细胞中的供体细胞，随后可用于细胞替代疗法 退行性疾病，如老年性黄斑变性。