Biophysical Regulation of Axial Polarity During Regeneration

再生过程中轴向极性的生物物理调节

基本信息

批准号：
7807945
负责人：
Wendy Scott Beane
金额：
$ 5.05万
依托单位：
TUFTS UNIVERSITY MEDFORD
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2011-04-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): (1) Clinical applications arising from stem cell and regeneration research will require an understanding not only of de novo tissue synthesis but also of the patterning mechanisms regulating blastema morphology. This project seeks to elucidate the regulatory networks controlled by biophysical epigenetic changes which orient blastema polarity relative to existing tissues. Our hypothesis is that ion transport by the H,K ATPase transporter and gap junctional communication (GJC) together regulate anterior-posterior polarity during planarian regeneration upstream of traditional signaling. Aim 1 is to identify and characterize transcriptional targets downstream of ion transport, through microarray analyses of regenerating worms inhibited for H,K ATPase (completed) and GJC activity. Candidates will be selected using specific criteria, validated by expression analyses, and functionally characterized following RNAi knockdown using in situ hybridization and molecular marker analyses of phenotypes. Aim 2 is to build and test models which outline the mechanisms ion transporters use to control axial identity in regenerating tissue. A preliminary transcriptional network downstream of ion transport will be generated by identifying network nodes, using quantitative PCR to test how RNAi knockdown of a single candidate affects the expression of all other candidates. Hypotheses predicted by this network will then be generated and tested, to elucidate the transcriptional and signaling response to the activity of the implicated ion transporters during the establishment of regenerative polarity. The long-term goal is to compile and validate a systems-level regeneration network model integrating biophysical and biochemical signaling. This has the potential for use as a predictive model of morphogenesis to aid in the translation of basic regeneration research into practical biomedical therapies. (2) The promise of stem cell and regeneration research is ultimately to replace lost or damaged tissues, organs and limbs in patients suffering from injury, aging or cancer. Although the regeneration of new tissue is critical, for that tissue to be functional, it must be properly patterned in relationship to the existing tissues: This research aims to discover how new and old tissues communicate to regulate this pattering, by learning how regenerating flatworms determine whether to form a new head and/or tail following amputation.

描述（由申请人提供）：（1）干细胞和再生研究产生的临床应用不仅需要了解从头组织合成，还需要了解调节胚基形态的模式机制。该项目旨在阐明由生物物理表观遗传变化控制的调控网络，这些变化确定胚基相对于现有组织的极性。我们的假设是，H,K ATP 酶转运蛋白的离子转运和间隙连接通讯 (GJC) 共同调节涡虫再生过程中传统信号传导的前后极性。目标 1 是通过对 H、K ATP 酶（已完成）和 GJC 活性抑制的再生蠕虫进行微阵列分析，识别和表征离子转运下游的转录靶标。将使用特定标准选择候选者，通过表达分析进行验证，并在 RNAi 敲低后使用原位杂交和表型分子标记分析进行功能表征。目标 2 是建立和测试模型，概述离子转运蛋白用于控制再生组织中的轴向同一性的机制。通过识别网络节点，使用定量 PCR 测试单个候选物的 RNAi 敲低如何影响所有其他候选物的表达，将生成离子传输下游的初步转录网络。然后将生成并测试该网络预测的假设，以阐明在再生极性建立过程中对相关离子转运蛋白活性的转录和信号反应。长期目标是编译和验证集成生物物理和生化信号传导的系统级再生网络模型。这有可能用作形态发生的预测模型，以帮助将基础再生研究转化为实际的生物医学疗法。 (2) 干细胞和再生研究的前景最终是替代遭受损伤、衰老或癌症的患者丢失或受损的组织、器官和四肢。尽管新组织的再生至关重要，但要使该组织发挥功能，它必须与现有组织建立适当的关系：这项研究旨在通过了解再生扁虫如何确定新组织和旧组织如何沟通来调节这种模式。截肢后是否形成新的头部和/或尾部。