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: 9913378
• 负责人: Corinne L. Fairchild
• 金额: $ 5.89万
• 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 2021-01-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9913378
• 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 Cone; Role; Science; 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; stem cells; 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.

项目摘要 正常的视觉功能依赖于视网膜中所有类型的细胞以正确的比例产生， 正确的发展时间。然而，我们对神经祖细胞如何决定哪种细胞 在胚胎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 从一个共同的视网膜祖细胞池中产生，随着时间的推移， 视网膜细胞类型的刻板顺序。但是，我们不知道控制能力的机制 过渡，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 微小RNA（miRNAs），短的单链核酸分子，负调控基因， 表达，是视网膜祖细胞从产生早期出生的视网膜细胞过渡到产生晚期出生的视网膜细胞所必需的。 类型;但是，我们不知道他们的行动机制。我的初步数据表明，miRNAs调节细胞 视网膜祖细胞的周期动力学，并导致我的假设，即miRNA介导的祖细胞能力 通过控制细胞周期长度。在这个建议中，我将通过以下目标来测试我的假设： 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：确定视网膜祖细胞周期长度如何影响细胞命运。 目的3：确定miRNA介导的细胞周期动力学变化是否介导鸡的发育 高灵敏度区域。 我将使用体内和体外技术的组合，包括流式细胞术，视网膜实时成像， 外植体，RNA测序和鸡胚电穿孔来回答我的研究问题。这将 大大扩展了我的技术技能，这将帮助我成为一个更具竞争力的候选人的教师 位置除了扩展我的技术技能外，在我的研究培训计划中，我还强调了软技能 作为一名学术研究人员，我需要召集一群经验丰富的导师， 提供以下类别的指导：职业发展，教学，资助写作和科学 外展加州大学戴维斯分校提供了一个非常协作的环境，并获得一流的核心设施;因此，它是 这是一个理想的环境，可以让我进行实验，并提高我的职业发展技能。 成功完成这里概述的项目将提供关键的洞察机制，控制 神经祖细胞命运的决定。这些信息对于建立有效的协议以生成 来自干细胞的供体细胞，其随后可用于细胞替代疗法以治疗 老年性黄斑变性等退行性疾病。