Probing the nanoscale organisation and multivalency of cell surface receptors: DNA origami nanoarrays for cellular studies with single-molecule control
Probing the nanoscale organisation and multivalency of cell surface receptors: DNA origami nanoarrays for cellular studies with single-molecule control
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DOI：
10.1039/c9fd00023b
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发表时间：
2019-10-01
影响因子：
3.4
通讯作者：
Palma, Matteo
中科院分区：
文献类型：
--
作者：
Hawkes, William;Huang, Da;Palma, Matteo
受体配体的纳米级组织已成为研究细胞表面受体聚集行为的重要途径。到目前为止，通过不同的纳米修饰策略制备的仿生底物已被应用于研究特定的整合素和细胞类型，但没有进行多价控制。在这里，我们利用DNA折纸超越现有方法的限制，制备纳米阵列来研究不同的细胞黏附过程，具有纳米级的空间分辨率和单分子控制。值得注意的是，DNA纳米结构能够以高度可定制的方式展示受体配体，具有可修改的参数，包括配体数量、配体间距以及最重要的多价性。为了测试系统的适应性和稳健性，我们将其与聚焦离子束和电子束光刻纳米刻蚀相结合，以另外控制折纸结构(即受体簇)之间的距离。此外，我们还演示了如何使用该平台来询问两个不同的生物学问题：(1)整合素和生长因子受体在癌细胞扩散中的协同作用；(2)整合素聚集在心肌细胞黏附和成熟中的作用。因此，我们发现了以前未知的不同整合素的聚集行为，进一步概述了这种可定制平台对于未来在纳米尺度上研究特定受体组织的重要性。
Nanoscale organisation of receptor ligands has become an important approach to study the clustering behaviour of cell-surface receptors. Biomimetic substrates fabricated via different nanopatterning strategies have so far been applied to investigate specific integrins and cell types, but without multivalent control. Here we use DNA origami to surpass the limits of current approaches and fabricate nanoarrays to study different cell adhesion processes, with nanoscale spatial resolution and single-molecule control. Notably, DNA nanostructures enable the display of receptor ligands in a highly customisable manner, with modifiable parameters including ligand number, ligand spacing and most importantly, multivalency. To test the adaptability and robustness of the system we combined it with focused ion beam and electron-beam lithography nanopatterning to additionally control the distance between the origami structures (i.e. receptor clusters). Moreover, we demonstrate how the platform can be used to interrogate two different biological questions: (1) the cooperative effect of integrin and growth factor receptor in cancer cell spreading, and (2) the role of integrin clustering in cardiomyocyte adhesion and maturation. Thereby we find previously unknown clustering behaviour of different integrins, further outlining the importance for such customisable platforms for future investigations of specific receptor organisation at the nanoscale.