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布置主要器官的复杂局限性结构。这种运动的例子是胃肠作用，它会产生肠道和神经造成脊柱和弦。内陷是一种形态发生的运动，在此过程中，细胞板扣（沿着神经术的一条线，沿着胃胃的圆圈）。内陷异常导致诸如脊柱裂之类的疾病，影响约1000分之一的活产。细胞表在产生分支器官（肺，肾脏，血管）或癌症期间肿瘤会导致新血管生长以提供营养时，细胞表也会经历。在内陷过程中，一组细胞必须表达与周围细胞不同的基因。已经在胚胎中广泛研究了内陷，并提出了几种理论来解释细胞片的起伏的机制。但是，由于胚胎中的所有细胞都经历了定义的基因表达序列，因此很难知道是什么原因导致了内幕的原因，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基因集。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。微图技术和微流体技术使我们能够以微米精度选择性处理细胞。通过结合两组技术，我将设计一个实验系统，该系统将使我能够在单层中强迫一部分细胞表达我选择的基因。另外，将将细胞培养在可以变形的软底物上。该系统将使我能够直接测试提出的每种不同理论。对于每个提出的理论，我将通过迫使单层中的一条细胞条纹来重复提出的运动，以承担该理论假设的运动类型是造成不败的原因。这将通过强制基因表达或化学处理来完成。然后，我只需在24小时内观察细胞板，看看它是否令人垂涎。一旦我发现哪些治疗引起了内陷，我将检查发挥作用的机械力。该项目将使我们能够更好地理解不败的机制，并鉴定可在癌症进展过程中限制其旨在抑制其的蛋白质。

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