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: 8463567
• 负责人: Derek John Stefanik
• 金额: $ 2.95万
• 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/8463567
• 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; 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; public health relevance; regenerative; small molecule; transcriptome sequencing

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.

描述（由申请人提供）：人类的再生能力有限，该组织因受伤或疾病而丢失。对可能发生再生的过程有更大的了解可以大大改善人体器官和脊髓损伤，退化性疾病以及数字或四肢丧失的医学方法。 这项拟议的研究将研究简单的动物模型，海葵，nematostella vectensis，在再生过程中单个发育信号通路的功能。具体而言，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具有活性，因此在胚胎发生和再生过程中，Nematostella似乎都使用类似的途径。综上所述，这导致了以下假设：wnt/ss-catenin途径在胚胎胚胎发生过程中驱动口腔焦点（O/A）规格的Wnt/SS-catenin途径也可以在再生过程中驱动O/A同一性。该假设产生了几个预测：1）在再生中，SS-catenin将定位于原住民组织中的细胞质并转移到口腔组织中的细胞核2）再生组织的轴向认同将由SS-Catenin的细胞内定位控制； 3）通过SS-catenin发出信号会激活驱动再生组织命运规范的转录程序。拟议的研究的总体目标是确定Wnt/SS-catenin信号传导在使用以下特定目的完成以下特定目的1：确定口服组织和异常组织中的SS-catenin的细胞内定位，确定SS-Catenin的细胞内定位，使用以下特定目的1：确定使用口服和异常组织抑制sss-catenin paterin patenib来确定SS-catenin的细胞内定位。 具体目的2：确定SS-catenin的核易位是否需要和/或足以通过将Wnt/SS-catenin信号与RNAi和小型途径成分的小分子抑制剂扰动，以抑制和/或诱导SS-CATENIN的SS-CATENIN的核转移。 特定目的3：在再生期间，在口腔和可靠组织的产生中鉴定由SS-catenin信号调节的基因。在扰动Wnt/SS-catenin信号传导后，将通过原位杂交和扰动与非扰动（对照）厌氧（对照）的转录组对转录组的高吞吐量进行调节。

项目成果

Production of a reference transcriptome and transcriptomic database (EdwardsiellaBase) for the lined sea anemone, Edwardsiella lineata, a parasitic cnidarian.

• DOI: 10.1186/1471-2164-15-71
• 发表时间: 2014-01-28
• 期刊: BMC genomics
• 影响因子: 4.4
• 作者: Stefanik DJ;Lubinski TJ;Granger BR;Byrd AL;Reitzel AM;DeFilippo L;Lorenc A;Finnerty JR
• 通讯作者: Finnerty JR

Collecting, rearing, spawning and inducing regeneration of the starlet sea anemone, Nematostella vectensis.

收集、饲养、产卵并诱导新生海葵 Nematostella vectensis。

• DOI: 10.1038/nprot.2013.044
• 发表时间: 2013
• 期刊: Nature protocols
• 影响因子: 14.8
• 作者: Stefanik,DerekJ;Friedman,LaurenE;Finnerty,JohnR
• 通讯作者: Finnerty,JohnR

Induction of canonical Wnt signaling by alsterpaullone is sufficient for oral tissue fate during regeneration and embryogenesis in Nematostella vectensis.

• DOI: 10.1002/dvdy.22774
• 发表时间: 2011-12
• 期刊: Developmental dynamics : an official publication of the American Association of Anatomists
• 作者: Trevino M;Stefanik DJ;Rodriguez R;Harmon S;Burton PM
• 通讯作者: Burton PM