A functional systems biology approach to investigating the role of Wnt/beta-caten

研究 Wnt/β-caten 作用的功能系统生物学方法

• 批准号: 8007571
• 负责人: Derek John Stefanik
• 金额: $ 2.9万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8007571
• 关键词: Adult; Anemone; Animal Model; Animals; Biological Models; Cell Nucleus; Cnidaria; Complex; Cytoplasm; Degenerative Disorder; Development; Developmental Biology; Digit structure; Disease; Embryo; Embryonic Development; Exhibits; Gene Expression Profile; Genes; Goals; Human; In Situ Hybridization; Injury; Knowledge; Laboratories; Laboratory culture; Learning; Limb structure; Liver; Medical; Molecular; Natural regeneration; Newts; Nuclear Translocation; Oral; Organ; Paralysed; Pathway interactions; Pattern; Planarians; Process; Production; Public Health; RNA Interference; Regulation; Research; Role; Sea Anemones; Signal Pathway; Signal Transduction; Specific qualifier value; Spinal cord injury; Structure; Systems Biology; Tissues; To specify; Ursidae Family; Vertebrates; Zebrafish; improved; inhibitor/antagonist; oral tissue; polyclonal antibody; programs; regenerative; small molecule

DESCRIPTION (provided by applicant): Humans have a limited ability to regenerate tissue that is lost due to injury or disease. A greater understanding of the processes by which regeneration can occur could substantially improve medical approaches toward human organ and spinal cord injury, degenerative disease and loss of digits or limbs. This proposed research will investigate the function of a single developmental signaling pathway during regeneration in a simple animal model, the sea anemone, Nematostella vectensis. Specifically, Wnt/ss-catenin signaling will be investigated. Since key embryonic signaling pathways are active in regenerating tissue, it seems quite likely that Nematostella utilizes a similar suite of pathways during both embryogenesis and regeneration. Taken together, this leads to the hypothesis that the Wnt/ss-catenin pathway, which drives oral-aboral (O/A) specification during embryogenesis of Nematostella, also drives O/A identity during regeneration. Several predictions arise from this hypothesis: 1) During regeneration ss-catenin will be localized to the cytoplasm in aboral tissue and translocated to the nucleus in oral tissue 2) The axial identity of regenerating tissue will be controlled by the intracellular localization of ss-catenin; and 3) That signaling through ss-catenin activates transcriptional programs that drive fate specification of regenerating tissue. The overall goal of the proposed research is to determine the role of Wnt/ss-catenin signaling in axial patterning of regenerating tissue in the sea anemone Nematostella by completing the following specific aims: Specific Aim 1: To determine the intracellular localization of ss-catenin during regeneration of oral and aboral tissue using a ss-catenin polyclonal antibody. Specific Aim 2: To determine whether nuclear translocation of ss-catenin is necessary and/or sufficient to specify O/A identity in regenerating tissue by perturbing Wnt/ss-catenin signaling with RNAi and small molecule inhibitors of pathway components to inhibit and/or induce nuclear translocation of ss-catenin during regeneration. Specific Aim 3: To identify genes regulated by ss-catenin signaling in the production of oral and aboral tissue during regeneration. Following perturbation of Wnt/ss-catenin signaling, regulation of Wnt/ss-catenin responsive genes will be accessed via in situ hybridization and high throughput sequencing of the transcriptome in perturbed vs. non-perturbed (control) anemones. PUBLIC HEALTH RELEVANCE: The proposed project will expand our understanding of the molecular mechanisms underlying regeneration. Increased knowledge of how organs and tissue regenerate bears direct relevance to public health issues such as human organ and spinal cord injury, degenerative disease, and loss of digits or limbs.

描述（由申请人提供）：人类再生因受伤或疾病而损失的组织的能力有限。更好地了解再生发生的过程可以大大改善针对人体器官和脊髓损伤、退行性疾病以及手指或四肢丧失的医疗方法。 这项拟议的研究将研究简单动物模型海葵 Nematostella vectensis 再生过程中单一发育信号通路的功能。具体来说，将研究 Wnt/ss-catenin 信号传导。由于关键的胚胎信号通路在再生组织中很活跃，因此线虫似乎很可能在胚胎发生和再生过程中利用了一套类似的通路。综上所述，这导致了这样的假设：Wnt/ss-连环蛋白途径在线虫胚胎发生过程中驱动口-口（O/A）规范，也在再生过程中驱动O/A身份。这一假设产生了几个预测：1) 在再生过程中，ss-catenin 将定位于口腔组织的细胞质，并易位至口腔组织的细胞核。 2) 再生组织的轴向特性将由 ss-catenin 的细胞内定位控制。 ; 3）通过单链蛋白发出的信号激活转录程序，驱动再生组织的命运规范。本研究的总体目标是通过完成以下具体目标来确定 Wnt/ss-catenin 信号在海葵 Nematostella 再生组织轴向模式中的作用： 具体目标 1：确定 ss-catenin 的细胞内定位在使用 ss-catenin 多克隆抗体进行口腔和口腔组织再生过程中。 具体目标 2：确定 ss-catenin 的核转位对于指定再生组织中的 O/A 身份是否是必要和/或充分的，方法是使用 RNAi 和途径成分的小分子抑制剂扰乱 Wnt/ss-catenin 信号传导来抑制和/或在再生过程中诱导 ss-catenin 的核转位。 具体目标 3：鉴定再生过程中口腔和口腔组织生成过程中受 ss-catenin 信号传导调节的基因。在扰动 Wnt/ss-catenin 信号传导后，将通过原位杂交和扰动与未扰动（对照）海葵转录组的高通量测序来了解 Wnt/ss-catenin 响应基因的调节。 公共健康相关性：拟议的项目将扩大我们对再生分子机制的理解。对器官和组织如何再生的了解的增加与人体器官和脊髓损伤、退行性疾病以及手指或四肢丧失等公共卫生问题直接相关。