| 研究課題/領域番号 |
20K05321
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| 研究機関 | 金沢大学 |
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研究代表者 |
YURTSEVER AYHAN 金沢大学, ナノ生命科学研究所, 特任助教 (00761529)
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| 研究期間 (年度) |
2020-04-01 – 2024-03-31
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| キーワード | exosomes / Atomic force Microscopy / Nanomechanics / Cells / 3D-AFM |
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| 研究実績の概要 |
We applied 3D-AFM to assess the structural and nanomechanical properties of exosomes released from different types of cells in a physiologically
relevant 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.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Research progress is sufficient. We achieved most of our previously planned research objectives. We determined the structural and mechanical characteristic of normal and cancer-derived exosomes for different cell types.
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| 今後の研究の推進方策 |
The biochemical composition of exosomes will be determined using functionalized AFM probes with biomolecules specific to exosome surface receptors and proteins Furthermore, we will perform nanomechnaical measurements of exosomes from different cell types (PC-9, PC-9 BrM4 and WM266.4, WM266.4 BrM3 cells).
The internal composition content of exosomes, such as miRNA and DNAs, will be determined by high resolution AFM imaging and force mapping by rupturing the lipid membrane of exosomes.
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| 次年度使用額が生じた理由 |
I had planned to attend domestic and international conferences, but due to corona many conferences were canceled or went online. Because of that, I couldn't go on a business trip, so the plan has changed; some funds will be used in the next fiscal year.
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