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Diversity Supplement -- Vertebrate Photoreceptor Development and Regeneration

多样性补充——脊椎动物感光器的发育和再生

基本信息

批准号：
10650081
负责人：
Ann C Morris
金额：
$ 8.42万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10650081
关键词：
Adoption Animal Model Biochemical Biology Blindness Cell Differentiation process Cell Transplantation Cells Clustered Regularly Interspaced Short Palindromic Repeats Cone Data Defect Development Developmental Biology Disease Embryo Event Evolution Eye diseases Family Gene Targeting Genetic Genetic Diseases Genetic Transcription Goals Health Homologous Gene Human Image In Vitro Inherited Injury Laboratories Mediating Microphthalmos Modeling Molecular Molecular Genetics Molecular Target Morphogenesis Morphology Mutation Natural regeneration Nerve Regeneration Neurons Optics Pathway interactions Photoreceptors Protocols documentation Retina Retinal Cone Retinal Degeneration Retinal Dystrophy Retinitis Pigmentosa Rod Role SOX11 gene SOX4 gene Signal Pathway Signal Transduction Suggestion System Therapeutic Time Transcription Repressor Transgenic Organisms Transplantation Vascular Endothelial Growth Factors Vertebrate Photoreceptors Visual Visual impairment Visual system structure Work Zebrafish base cellular imaging clinical application dosage experimental study gene regulatory network gene therapy genetic approach genome editing imaging modality improved in vivo loss of function member mouse genome mutant neurogenesis notch protein novel optic cup parent grant photoreceptor degeneration precursor cell response retinal neuron retinal progenitor cell retinal rods stem cell differentiation stem cell proliferation therapy development transcription factor transplantation therapy

项目摘要

PROJECT SUMMARY Inherited retinal dystrophies such as retinitis pigmentosa (RP) are a leading cause of blindness for which there is currently no cure. One potential treatment for retinal degenerative diseases is cell- based transplantation therapy, whereby precursor cells are transplanted into the diseased eye to replace the lost photoreceptors. While this is an exciting possibility, several challenges to the implementation of transplantation therapies must be overcome, including the inefficient integration of photoreceptor precursors into the recipient retina, and the difficulty of obtaining sufficient numbers of photoreceptor precursors for clinical application. To improve protocols for the in vitro culture of such cells, we must have a better understanding of the transcriptional networks that promote specification of photoreceptor precursors. One of the long-term goals of our laboratory is to contribute to these efforts by studying photoreceptor development and regeneration in the zebrafish. The zebrafish is especially useful for studying photoreceptor biology, because its retina contains numerous cone subtypes in addition to rods. Furthermore, the zebrafish retina is able to regenerate neurons in response to experimental damage. The experiments described in this proposal will define the role of three transcription factors -- Sox, Sox11, and Her9 -- during retinal neurogenesis through the application of gene targeting and molecular genetic approaches. Our specific aims are as follows: Specific Aim I: Determine the role of Sox4 and Sox11 in photoreceptor differentiation. Using newly generated loss-of-function mutants, we will 1) determine whether there is a dosage effect of Sox4/11 activity on rod photoreceptor number; 2) use state-of-the-art time-lapse imaging to determine when and where Sox4 expression is required to achieve proper rod photoreceptor development; 3) determine precisely how Sox4/11 regulate Hh and Bmp signaling; and 4) identify the molecular targets of Sox4/11. Specific Aim II: Determine the cause and extent of photoreceptor defects in the zebrafish her9 mutant. In this aim, we will 1) determine the mechanism for photoreceptor defects in her9 mutants; 2) determine whether loss of Her9 impairs vision; 3) identify the upstream signaling pathway(s) that regulate her9 expression in the retina; and 4) determine whether Her9-mediated VEGF expression in the avascular zebrafish retina regulates retinal progenitor cell proliferation and differentiation. Completion of our proposal will bridge important gaps in our understanding of the molecular mechanisms of vertebrate photoreceptor differentiation, and reveal underlying principles relevant to the development of approaches for the treatment of human retinal degenerative disease.

项目总结遗传性视网膜营养不良，如视网膜色素变性(RP)，是导致失明的主要原因目前还没有治愈方法。视网膜退行性疾病的一种潜在治疗方法是细胞- 以移植为基础的疗法，将前体细胞移植到患病的眼睛中以更换丢失的光感受器。虽然这是一种令人兴奋的可能性，但必须克服移植疗法的实施，包括低效地整合光感受器前体进入受体视网膜，以及获得足够数量的临床应用的光感受器前体。为了改进这类细胞的体外培养方案细胞，我们必须对促进规范的转录网络有更好的了解光感受器前体。我们实验室的长期目标之一是为实现这些目标做出贡献通过研究斑马鱼的光感受器发育和再生的努力。斑马鱼是对研究光感受器生物学特别有用，因为它的视网膜含有大量的视锥除棒材外，还有其他亚型。此外，斑马鱼的视网膜能够在对实验性损伤的反应。本提案中描述的实验将定义三种转录因子--SOX、SOX11和Her9--通过基因打靶和分子遗传学方法的应用。我们的具体目标如下：具体目的I：确定Sox4和Sox11在光感受器分化中的作用。使用新的产生的功能丧失突变体，我们将1)确定Sox4/11是否有剂量效应视杆感光细胞活动数目；2)使用最先进的时间推移成像来确定以及需要Sox4表达才能实现杆状感光细胞的正常发育；3) 准确确定Sox4/11如何调节HH和BMP信号；以及4)识别分子 SOX4/11的靶点。特定目标II：确定光感受器缺陷的原因和程度斑马鱼HER9突变体。在这个目标中，我们将1)确定光感受器缺陷的机制 Her9突变体；2)确定丢失Her9是否会损害视力；3)确定上游信号调节视网膜中her9表达的通路(S)；以及4)决定her9是否介导血管内皮生长因子在斑马鱼视网膜中的表达调节视网膜前体细胞的增殖和差异化。我们的建议的完成将弥合我们对脊椎动物光感受器分化的分子机制，并揭示潜在的原理与开发治疗人类视网膜退行性疾病的方法有关。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Sox11 is required to maintain proper levels of Hedgehog signaling during vertebrate ocular morphogenesis.

DOI：
10.1371/journal.pgen.1004491
发表时间：
2014-07
期刊：
PLoS genetics
影响因子：
4.5
作者：
Pillai-Kastoori L;Wen W;Wilson SG;Strachan E;Lo-Castro A;Fichera M;Musumeci SA;Lehmann OJ;Morris AC
通讯作者：
Morris AC

Keeping an eye on SOXC proteins.

DOI：
10.1002/dvdy.24235
发表时间：
2015-03
期刊：
Developmental dynamics : an official publication of the American Association of Anatomists
影响因子：
0
作者：
Pillai-Kastoori L;Wen W;Morris AC
通讯作者：
Morris AC

Generation of a zebrafish knock-in line expressing MYC-tagged Sox11a using CRISPR/Cas9 genome editing.