The molecular basis of crosstalk between integrin-mediated cell-matrix adhesions and the microtubule network.

整合素介导的细胞基质粘附和微管网络之间串扰的分子基础。

• 批准号: BB/N007336/1
• 负责人: Benjamin Goult
• 金额: $ 46.9万
• 依托单位: University of Kent
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN007336%2F1
• 关键词: molecular; basis; crosstalk; between; integrin

All cells in the human body are held in the correct place via adhesion to neighbouring cells, and to a dense meshwork of proteins that surround cells called the extracellular matrix (ECM). Cell attachment to the ECM is important for cell proliferation, survival and migration during embryonic development, and in adult organisms for processes such as blood clotting and wound healing. Cell-matrix adhesion is mediated by a family of proteins, the integrins, that are found on the cells surface that act as receptors that grab hold of ECM proteins.Integrins are organised into specialised structures called focal adhesions (FAs) - sticky feet - and cell migration requires the coordinated assembly and disassembly of FAs coupled to force exerted by the cells contractile machinery which enables the cell to pull itself forwards. While cell migration is essential for the development of multicellular organisms, it must be tightly controlled - cancer metastasis is a product of uncontrolled cell migration. From our previous research and that of other laboratories, the assembly of FAs is now fairly well understood, and an intracellular protein called talin has been shown to be essential for FA assembly as it binds to integrins and couples them to the cells' contractile machinery. However, the mechanisms underlying the disassembly of FAs are less clear. It is now well recognised that recruitment of intracellular structures called microtubules to FAs enhances their turnover and increases the speed at which cells migrate, but what directs microtubules to FAs has remained elusive. Excitingly, we have discovered a novel link between talin and a protein called Kank1 that is involved in stabilising microtubules. The hypothesis that we would like to test is that the interaction between talin and Kank1 directs microtubules to adhesion sites and in doing so leads to FA disassembly and consequently increased cell migration.Drugs that disrupt microtubules are commonly used to kill cancer cells, but have severe side effects because they also kill normal cells. However, if we can leave the microtubule network intact, but specifically prevent microtubule recruitment to FAs, this should inhibit FA turnover and suppress cell migration, and therefore offer a new way to treat cancer metastasis. Until we characterise how the talin:Kank1 interaction works, and determine the precise atomic structure of the binding sites where this linkage is made, we cannot design drugs to prevent the talin-dependent recruitment of microtubules to FAs that enhances the invasiveness of cancer cells. The Aim of this project is to establish how talin and Kank1 interact so as to design mutations in the Kank1 binding site that prevent the interaction. We will use a combination of biochemical and structural approaches to obtain this information, approaches that are well established in the applicant's laboratory. The effect of such mutations on cell migration will then be tested in collaboration with Prof. Anna Akhmanova (Utrecht).Ultimately this study could lead to the identification of new strategies for the design and development of drugs that inhibit microtubule recruitment to adhesion sites, and thus prevent cancer growth and spread in humans.

人体中的所有细胞都通过粘附到相邻细胞以及细胞周围称为细胞外基质（ECM）的密集蛋白质网络而保持在正确的位置。细胞附着于ECM对于胚胎发育期间的细胞增殖、存活和迁移以及在成体生物体中对于诸如血液凝固和伤口愈合的过程是重要的。细胞与基质的粘附是由一个蛋白质家族--整合素介导的，整合素存在于细胞表面，作为抓住ECM蛋白的受体。整合素被组织成称为粘着斑（focal adhesion，FA）的特殊结构--粘足--细胞迁移需要FA的协调组装和拆卸，再加上细胞收缩机制施加的力，使细胞能够向前拉动自己。虽然细胞迁移对于多细胞生物体的发育至关重要，但必须严格控制-癌症转移是不受控制的细胞迁移的产物。从我们以前的研究和其他实验室的研究中，FA的组装现在已经相当好地理解，并且一种称为talin的细胞内蛋白质已被证明是FA组装所必需的，因为它与整合素结合并将它们与细胞的收缩机制偶联。然而，潜在的机制拆卸的脂肪酸不太清楚。现在人们已经认识到，细胞内结构（称为微管）向FA的募集增强了它们的周转，并增加了细胞迁移的速度，但什么引导微管向FA仍然难以捉摸。令人兴奋的是，我们发现了talin和一种名为Kank 1的蛋白质之间的新联系，该蛋白质参与稳定微管。我们想要验证的假设是，talin和Kank 1之间的相互作用将微管引导到粘附位点，这样做会导致FA分解，从而增加细胞迁移。破坏微管的药物通常用于杀死癌细胞，但由于它们也会杀死正常细胞，因此具有严重的副作用。然而，如果我们能保持微管网络的完整性，但特异性地阻止微管募集到FA，这将抑制FA周转并抑制细胞迁移，从而提供一种治疗癌症转移的新方法。在我们弄清talin：Kank 1相互作用的原理，并确定这种连接的结合位点的精确原子结构之前，我们无法设计药物来阻止微管依赖于talin的FA募集，从而增强癌细胞的侵袭力。本项目的目的是确定talin和Kank 1如何相互作用，以便在Kank 1结合位点设计突变，阻止相互作用。我们将使用生物化学和结构方法的组合来获得这些信息，这些方法在申请人的实验室中得到了很好的建立。这些突变对细胞迁移的影响将与安娜·阿赫洛娃教授（乌得勒支）合作进行测试。最终，这项研究可能导致确定新的策略，用于设计和开发抑制微管募集到粘附位点的药物，从而防止癌症在人类中的生长和扩散。

项目成果

Bridging the Gap

• DOI: 10.1007/bf03311768
• 发表时间: 2014
• 期刊: InPharma
• 作者: T. van Amelsvoort

Talin mechanosensitivity is modulated by a direct interaction with cyclin-dependent kinase-1.

• DOI: 10.1016/j.jbc.2021.100837
• 发表时间: 2021-07
• 期刊: The Journal of biological chemistry
• 作者: Gough RE;Jones MC;Zacharchenko T;Le S;Yu M;Jacquemet G;Muench SP;Yan J;Humphries JD;Jørgensen C;Humphries MJ;Goult BT
• 通讯作者: Goult BT

Talin rod domain-containing protein 1 (TLNRD1) is a novel actin-bundling protein which promotes filopodia formation.

• DOI: 10.1083/jcb.202005214
• 发表时间: 2021-09-06
• 期刊: The Journal of cell biology
• 作者: Cowell AR;Jacquemet G;Singh AK;Varela L;Nylund AS;Ammon YC;Brown DG;Akhmanova A;Ivaska J;Goult BT
• 通讯作者: Goult BT

Cancer associated talin point mutations disorganise cell adhesion and migration

癌症相关的talin点突变扰乱细胞粘附和迁移

• DOI: 10.1101/2020.03.25.008193
• 发表时间: 2020
• 作者: Azizi L