Genetic Analysis of Cell Invasion through Basement Membranes

细胞通过基底膜侵袭的遗传分析

基本信息

批准号：
7912123
负责人：
David R Sherwood
金额：
$ 24.22万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-11 至 2011-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Our long-term goal is to elucidate the genetic networks that direct cell invasion through basement membranes, the dense, sheet-like extracellular matrix that surrounds most tissues. The mechanisms that cells employ to cross basement membranes in vivo remain poorly understood, as these invasions most often occur in complex environments that are difficult to study. We are thus dissecting the process of anchor-cell (AC) invasion into the vulval epithelium in the visually and genetically accessible model organism Caenorhabiditis elegans. AC invasion involves: (1) the attachment of the AC to the basement membrane, (2) its polarization towards the basement membrane, (3) the generation and reception of a chemotactic signal(s) that stimulates invasion, (4) the precise removal of the basement membrane and (5) transit through the basement membrane. We have discovered a novel role for the netrin pathway in directing the polarization of the AC's invasive cellular processes towards the basement membrane. We have also identified a specific isoform of the C. elegans fos transcription factor, fos-1b, which inhibits AC invasion, perhaps by blocking fos-1a activity, an isoform that promotes basement membrane removal during AC invasion. Finally, we have conducted a pilot screen using a database generated from previous whole genome RNAi screens and identified five additional genes that promote AC invasion, four of which have not previously been implicated in regulating cell invasion. Integrating cellular, genetic, and molecular approaches, our proposed work will: 1) elucidate a new role for netrin signaling in polarizing an invasive cell, 2) determine the mechanisms by which fos-1b inhibits AC invasion, and 3) characterize the function of new genes identified in our RNAi database screen that specifically promote removal of the basement membrane during AC invasion. Cell invasions through basement membranes play crucial roles during normal development and are essential for leukocyte trafficking to sites of infection and injury. Uncontrolled cell- invasive activity is also associated with a number of deadly diseases, including cancer and rheumatoid arthritis. The proposed work will advance our understanding of the fundamental mechanisms controlling cell- invasive behavior and thus has a strong potential to lead to new treatment strategies for a number of human diseases associated with unregulated cell-invasive activity.

描述（由申请人提供）：我们的长期目标是阐明通过基底膜引导细胞侵袭细胞的遗传网络，地下膜是大多数组织周围的密集，类似细胞的细胞外基质。细胞在体内穿越基底膜的机制仍然很少了解，因为这些入侵通常发生在难以研究的复杂环境中。因此，我们正在视觉和遗传上可访问的模型生物秀丽隐杆线虫中的锚固细胞（AC）侵袭到外阴上皮的过程。 AC侵袭涉及：（1）AC附着在基底膜上，（2）其极化向地下膜，（3）趋化信号的产生和接收（S）（S）刺激侵袭，（4）（4）基底膜和（5）通过地下膜的交流。我们已经发现了Netrin途径在将AC侵入性细胞过程指向基底膜的极化中的新作用。我们还确定了秀丽隐杆线虫FOS转录因子FOS-1B的特异性同工型，该因子抑制AC侵袭，也许是通过阻断FOS-1A活性，这是一种在AC侵袭过程中促进基底膜去除的同工型。最后，我们使用了先前的整个基因组RNAi筛选产生的数据库进行了一个试验屏幕，并确定了五个促进AC侵袭的基因，其中四个以前没有与调节细胞侵袭有关。整合细胞，遗传和分子方法，我们的拟议工作将：1）阐明Netrin信号在极化A侵入性细胞中的新作用，2）确定FOS-1B抑制AC侵袭的机制，以及3）表征我们在RNAi数据库筛选中鉴定出的新基因的功能，这些筛查在我们的RNAi数据库中均促进了基础上的AC摄入量表，该机制是在AC Incesement exement exement exement exement exement exement exement exement exement mexembermbrane in Accasige of Acsement exement of Acsement of Acsement的函数。通过基底膜的细胞入侵在正常发育过程中起着至关重要的作用，对于白细胞运输到感染和损伤部位至关重要。不受控制的细胞浸润活性也与多种致命疾病有关，包括癌症和类风湿关节炎。拟议的工作将提高我们对控制细胞侵入性行为的基本机制的理解，因此具有强大的潜力，可以为许多与不受管制的细胞侵入性活性相关的许多人类疾病提供新的治疗策略。