Regulators of Contact Stimulation of Migration in Testis Nascent Myotubes

睾丸新生肌管迁移接触刺激的调节因子

• 批准号: 466788046
• 负责人: Dr. Maik Christian Bischoff
• 金额: --
• 依托单位: Department of Biology
• 依托单位国家: 德国
• 项目类别: WBP Fellowship
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/466788046?language=en
• 关键词: Regulators; Contact; Stimulation; Migration; Testis

Collective cell migration plays a crucial role in development, wound healing and metastasis. During my doctorate I established a new model for collective cellular motility, the Drosophila testis nascent myotube system. Using state-of-the-art live-cell imaging as well as pharmacological and genetic perturbation, I identified a new mechanism of contact-stimulated collective migration (CSM) in which a contact-dependent asymmetry of cell-matrix adhesion acts as a major switch to drive directional motion towards the free space, whereas contractile actin cables contribute to the integrity of the migrating muscle cell cluster. In the proposed project I will address how this intricate behaviour is regulated at the molecular level. Previously, it could be shown to be dependent of differential cell-cell contact dependent regulation of matrix-adhesion stability, apparently orchestrated by the Rho-family GTPases Rac2 and Cdc42. RBD-based biosensors will be created and applied to assess where Rac2 and Cdc42 are active on a subcellular level. Single-cell sequencing analysis of pre-migratory and migratory muscle cells will be applied, to assess which genes are upregulated and may be upstream of local GTPase activation. Subsequently, bioinformatically prioritized candidates will be targeted by RNAi and cell-specific CRISPR/CAS9 techniques. Preliminary results show the collective migration of testis nascent myotubes to depend on MMP2. Therefore, myotubes seem to use mechanics during development, known from invasive cancer cell motility. To fully assess the influence of the microenvironment that presumably has to be degraded, the composition and source-tissues of the ECM (extracellular matrix) will be addressed. The contribution of ECM-composition on migration and invasive dynamics will be investigated by using tissue specific RNAi in adjacent tissues.

集体细胞迁移在伤口的发育、愈合和转移中起着至关重要的作用。在我的博士学位期间，我建立了一个集体细胞运动的新模型，果蝇睾丸新生肌管系统。利用最先进的活细胞成像以及药理学和遗传干扰，我确定了一种接触刺激集体迁移（CSM）的新机制，其中细胞-基质粘附的接触依赖不对称充当主要开关，驱动向自由空间的定向运动，而收缩的肌动蛋白电缆有助于迁移肌肉细胞簇的完整性。在提议的项目中，我将讨论如何在分子水平上调节这种复杂的行为。以前，它可以被证明是依赖于差异细胞-细胞接触依赖性基质粘附稳定性的调节，显然是由rho家族GTPases Rac2和Cdc42精心策划的。基于rbd的生物传感器将被创建并应用于评估Rac2和Cdc42在亚细胞水平上的活性。将应用前迁移和迁移肌肉细胞的单细胞测序分析，以评估哪些基因上调，可能是局部GTPase激活的上游。随后，生物信息学上优先的候选基因将被RNAi和细胞特异性CRISPR/CAS9技术靶向。初步结果显示，睾丸新生肌管的集体迁移依赖于MMP2。因此，肌管在发育过程中似乎使用了从侵袭性癌细胞运动中得知的力学。为了充分评估可能必须被降解的微环境的影响，将讨论ECM（细胞外基质）的组成和来源组织。ecm组成对迁移和侵袭动力学的贡献将通过在邻近组织中使用组织特异性RNAi来研究。