A specific subset of RabGTPases controls cell surface exposure of MT1-MMP, extracellular matrix degradation and three-dimensional invasion of macrophages
A specific subset of RabGTPases controls cell surface exposure of MT1-MMP, extracellular matrix degradation and three-dimensional invasion of macrophages
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DOI：
10.1242/jcs.122358
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发表时间：
2013-07-01
影响因子：
4
通讯作者：
Linder, Stefan
中科院分区：
文献类型：
--
作者：
Wiesner, Christiane;El Azzouzi, Karim;Linder, Stefan
基质金属蛋白酶MT1-MMP在生理和病理环境如免疫细胞外渗或癌细胞转移中对侵袭性细胞迁移具有重要影响。表面相关的MT1-MMP能够切割细胞外基质的组分，这是蛋白水解侵入性迁移的先决条件。然而，目前的知识的分子机制，调节MT1-MMP运输和从细胞表面是有限的。我们已经确定了RabGT3家族的三个成员Rab5a、Rab8a和Rab14，作为原代人巨噬细胞中MT1-MMP运输和功能的关键调节因子。过表达和内源性形式都显示出与MT1-MMP阳性囊泡的显著共定位，而突变体构建体的表达以及siRNA诱导的敲低揭示了这些RabGTP酶在调节MT1-MMP表面暴露、MT1-MMP阳性囊泡与podosomes的接触、二维和三维的细胞外基质降解以及巨噬细胞的三维蛋白水解侵袭中至关重要。总的来说，我们的研究结果确定Rab5a，Rab8a和Rab14作为MT1-MMP运输和原代人巨噬细胞侵袭性迁移的主要调节因子，这可能是有前途的潜在靶点，用于操纵免疫细胞侵袭。
The matrix metalloproteinase MT1-MMP has a major impact on invasive cell migration in both physiological and pathological settings such as immune cell extravasation or metastasis of cancer cells. Surface-associated MT1-MMP is able to cleave components of the extracellular matrix, which is a prerequisite for proteolytic invasive migration. However, current knowledge on the molecular mechanisms that regulate MT1-MMP trafficking to and from the cell surface is limited. We have identified three members of the RabGTPase family, Rab5a, Rab8a and Rab14, as crucial regulators of MT1-MMP trafficking and function in primary human macrophages. Both overexpressed and endogenous forms show prominent colocalisation with MT1-MMP-positive vesicles, whereas expression of mutant constructs, as well as siRNA-induced knockdown, reveal that these RabGTPases are crucial in the regulation of MT1-MMP surface exposure, contact of MT1-MMP-positive vesicles with podosomes, extracellular matrix degradation in two and three dimensions, as well as three-dimensional proteolytic invasion of macrophages. Collectively, our results identify Rab5a, Rab8a and Rab14 as major regulators of MT1-MMP trafficking and invasive migration of primary human macrophages, which could be promising potential targets for manipulation of immune cell invasion.