Identification of early stage malignancy changes in the structural and biophysical proporties of exosomes by atomic force microscopy (AFM)-based nano-mechanical measurements

通过基于原子力显微镜（AFM）的纳米机械测量鉴定外泌体结构和生物物理特性的早期恶性肿瘤变化

• 批准号: 20K05321
• 负责人: YURTSEVER AYHAN
• 金额: $ 2.75万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2020
• 资助国家: 日本
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-20K05321/
• 关键词: exosomes; Atomic force Microscopy; Nanomechanics; Cells; 3D-AFM; Exosomes; Cell; AFM

We applied 3D-AFM to assess the structural and nanomechanical properties of exosomes released from different types of cells in a physiologicallyrelevant environment. We established A method for studying the interaction of molecules exposed on the exosomes by using AFM. Exploring the molecule exposed on the native exosome surface will deepen our understanding of the physiological function of exosomes in cancer progression. In addition, we performed AFM measurements to identify the exosomes in heterogeneous extracellular vesicles (EVs) and analysis their biophysical properties. There is no method that can isolate exosomes (EXOs) or microvesicles (MVs) without including other EVs. As a substitute for EXOs, intraluminal vesicles (ILVs), which are the origin of EXOs were isolated from cells. After separating EVs by the anion exchange column, we performed AFM nanomechanical characterization to determine the biophysical properties of exosomes and other EV subtypes.AFM nanomechanical measurements determined the rapture characteristic of EVs. We have found differences in their rapture mechanics, implying difference of membrane structure related to elastic fibers. Mass spectroscopic analyses were performed to determine their protein and lipid compositions.

我们应用3D-AFM来评估在生理相关环境中从不同类型细胞释放的外泌体的结构和纳米力学特性。建立了一种利用原子力显微镜研究外泌体表面分子相互作用的方法。探索暴露在天然外泌体表面的分子将加深我们对外泌体在癌症进展中的生理功能的理解。此外，我们进行了AFM测量，以确定在异质性细胞外囊泡（EV）中的外泌体，并分析其生物物理特性。没有一种方法可以在不包括其他EV的情况下分离外来体（EXO）或微泡（MV）。作为EXO的替代物，从细胞中分离作为EXO来源的腔内囊泡（ILV）。在用阴离子交换柱分离EV后，我们进行了AFM纳米力学表征，以确定exosomes和其他EV亚型的生物物理特性。AFM纳米力学测量确定了EV的断裂特性。我们发现它们的断裂机制不同，这意味着与弹性纤维有关的膜结构的差异。进行质谱分析以确定其蛋白质和脂质组成。

项目成果

Probing the structural and nanomechanical properties of bionanomaterials by three-dimensional atomic force microscopy (3D-AFM)

通过三维原子力显微镜 (3D-AFM) 探测生物纳米材料的结构和纳米力学特性

• 发表时间: 2022
• 作者: Onishi Hideto;Miyake Mikio;Shirai Hajime;Ayhan Yurtsever
• 通讯作者: Ayhan Yurtsever

Probing the Structural Details of Chitin Nanocrystal-Water Interfaces by Three‐Dimensional Atomic Force Microscopy (Small Methods 9/2022)

通过三维原子力显微镜探测甲壳素纳米晶体-水界面的结构细节（小方法 9/2022）

• DOI: 10.1002/smtd.202270054
• 发表时间: 2022
• 期刊: Small Methods
• 影响因子: 12.4
• 作者: Yurtsever Ayhan;Wang Pei‐Xi;Priante Fabio;Morais Jaques Ygor;Miyata Kazuki;MacLachlan Mark J.;Foster Adam S.;Fukuma Takeshi
• 通讯作者: Fukuma Takeshi

3D-AFM investigations of solvation structures on polysaccharide-based nanocrystals and self-assembled peptide nanostructures

多糖纳米晶体和自组装肽纳米结构溶剂化结构的 3D-AFM 研究

• 发表时间: 2022
• 作者: RANJBAR Sina;Satoshi Sumi;Sota Kambe;Kenji Tanabe;Hiroyuki Awano;Ayhan Yurtsever
• 通讯作者: Ayhan Yurtsever

Probing the structural details of cellulose and chitin nanocrystal-water interfaces by 3D-AFM

通过 3D-AFM 探测纤维素和甲壳素纳米晶体-水界面的结构细节

• 发表时间: 2022
• 作者: 佐々木 樹;孫 麗娜;黒澤 優;高橋 辰宏;硯里 善幸;A. Yurtsever,P. Wang,F. Priante,Y. M. Jaques,K. Miyazawa,K. Miyata,M. J,MacLachlan,A. S. Foster,T. Fukuma
• 通讯作者: A. Yurtsever,P. Wang,F. Priante,Y. M. Jaques,K. Miyazawa,K. Miyata,M. J,MacLachlan,A. S. Foster,T. Fukuma