Molecular and biophysical investigation of epithelial cell sheet invagination

上皮细胞片内陷的分子和生物物理研究

• 批准号: BB/F019769/1
• 负责人: Guillaume Charras
• 金额: $ 44.57万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FF019769%2F1
• 关键词: Molecular; biophysical; investigation; epithelial; cell

Early morphogenetic movements transform the new embryo from an amorphous ball of cells into a complex compartimentalised structure in which the main organs are laid out. Examples of such movements are gastrulation which creates the primitive gut and neurulation which creates the spinal chord. Invagination is a morphogenetic movement during which a cell sheet buckles (along a line in the case of neurulation and along a circle in the case of gastrulation). Abnormalities in invagination lead to conditions such as spina bifida that affect approximately 1 in 1000 live births. Cell sheets also undergo invagination during the creation of branched organs (lungs, kidneys, blood vessels) or during cancer when the tumour causes new blood vessels to grow to provide it with nutrients. During invagination, one group of cells has to express different genes than the surrounding cells. Invagination has been extensively studied in embryos and several theories have been proposed to explain the mechanism through which a cell sheet invaginates. However, since all cells in embryos undergo a defined sequence of gene expression, it is difficult to know what is responsible for invagination and what is just a side effect. All of the proposed theories for invagination require that one subset of the cell population express a different set of genes than the surrounding cells. Modern molecular biology techniques enable us to force cells to express a gene of choice. Microprinting techniques and microfluidic techniques enable us to selectively treat cells with micrometer precision. By combining both sets of techniques, I will devise an experimental system that will allow me to force a subset of cells within a monolayer to express a gene of my choice. In addition, the cells will be cultured on a soft substrate that they can deform. This system will enable me to test each of the different theories proposed for invagination directly. For each proposed theory, I will replicate the proposed movement by forcing a stripe of cells within the monolayer to undergo the type of movement that the theory hypothesizes is the cause of invagination. This will be done either through forced gene expression or through chemical treatment. Then, I will simply observe the cell sheet over a period of 24 hours and see if it invaginates. Once I have found which treatments give rise to invaginations, I will examine the mechanical forces at play. This project will enable us to better understand the mechanisms of invagination and identify proteins that can be targeted to inhibit it during cancer progression.

早期的形态发生运动将新胚胎从无定形的细胞球转变为复杂的区室化结构，其中布置了主要器官。这种运动的例子是原肠胚形成和神经形成，前者产生原始肠道，后者产生脊髓。内陷是一种形态发生运动，在此过程中细胞片发生弯曲（在神经形成的情况下沿着一条线，在原肠胚形成的情况下沿着一个圆圈）。内陷异常会导致脊柱裂等疾病，影响大约千分之一的活产儿。在分支器官（肺、肾、血管）形成过程中或在癌症过程中，当肿瘤导致新血管生长为其提供营养时，细胞片层也会发生内陷。在内陷期间，一组细胞必须表达与周围细胞不同的基因。内陷已在胚胎中得到广泛研究，并提出了几种理论来解释细胞片内陷的机制。然而，由于胚胎中的所有细胞都会经历确定的基因表达序列，因此很难知道什么是造成内陷的原因以及什么只是副作用。所有提出的内陷理论都要求细胞群的一个子集表达与周围细胞不同的一组基因。现代分子生物学技术使我们能够迫使细胞表达选择的基因。微印刷技术和微流体技术使我们能够以微米精度选择性地处理细胞。通过结合两组技术，我将设计一个实验系统，使我能够迫使单层内的细胞子集表达我选择的基因。此外，细胞将培养在可以变形的柔软基质上。该系统将使我能够直接测试针对内陷提出的每种不同理论。对于每个提出的理论，我将通过迫使单层内的细胞条纹经历该理论假设的导致内陷的运动类型来复制所提出的运动。这可以通过强制基因表达或通过化学处理来完成。然后，我会简单地观察细胞片24小时，看看它是否内陷。一旦我发现哪些治疗会引起内陷，我将检查起作用的机械力。该项目将使我们能够更好地了解内陷的机制，并识别在癌症进展过程中可以靶向抑制内陷的蛋白质。

项目成果

In vivo collective cell migration requires an LPAR2-dependent increase in tissue fluidity.

• DOI: 10.1083/jcb.201402093
• 发表时间: 2014-07-07
• 期刊: The Journal of cell biology
• 作者: Kuriyama S;Theveneau E;Benedetto A;Parsons M;Tanaka M;Charras G;Kabla A;Mayor R
• 通讯作者: Mayor R

Mechanisms of leading edge protrusion in interstitial migration.

• DOI: 10.1038/ncomms3896
• 发表时间: 2013
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Wilson, Kerry;Lewalle, Alexandre;Fritzsche, Marco;Thorogate, Richard;Duke, Tom;Charras, Guillaume
• 通讯作者: Charras, Guillaume

Cellular control of cortical actin nucleation.

• DOI: 10.1016/j.cub.2014.05.069
• 发表时间: 2014-07-21
• 期刊: Current biology : CB
• 作者: Bovellan M;Romeo Y;Biro M;Boden A;Chugh P;Yonis A;Vaghela M;Fritzsche M;Moulding D;Thorogate R;Jégou A;Thrasher AJ;Romet-Lemonne G;Roux PP;Paluch EK;Charras G
• 通讯作者: Charras G