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细胞外囊泡周围的功能性 corona 增强了血管生成、皮肤再生和免疫调节。

A functional corona around extracellular vesicles enhances angiogenesis, skin regeneration and immunomodulation.

机构信息

Cell Therapy Institute, Spinal Cord Injury and Tissue Regeneration Centre Salzburg (SCI-TReCS), Paracelsus Medical University (PMU), Salzburg, Austria.

Department of Biosciences, Paris Lodron University Salzburg, Salzburg, Austria.

出版信息

J Extracell Vesicles. 2022 Apr;11(4):e12207. doi: 10.1002/jev2.12207.

DOI:10.1002/jev2.12207
PMID:35398993
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8994701/
Abstract

Nanoparticles can acquire a plasma protein corona defining their biological identity. Corona functions were previously considered for cell-derived extracellular vesicles (EVs). Here we demonstrate that nano-sized EVs from therapy-grade human placental-expanded (PLX) stromal cells are surrounded by an imageable and functional protein corona when enriched with permissive technology. Scalable EV separation from cell-secreted soluble factors via tangential flow-filtration (TFF) and subtractive tandem mass-tag (TMT) proteomics revealed significant enrichment of predominantly immunomodulatory and proangiogenic proteins. Western blot, calcein-based flow cytometry, super-resolution and electron microscopy verified EV identity. PLX-EVs partly protected corona proteins from protease digestion. EVs significantly ameliorated human skin regeneration and angiogenesis in vivo, induced differential signalling in immune cells, and dose-dependently inhibited T cell proliferation in vitro. Corona removal by size-exclusion or ultracentrifugation abrogated angiogenesis. Re-establishing an artificial corona by cloaking EVs with fluorescent albumin as a model protein or defined proangiogenic factors was depicted by super-resolution microscopy, electron microscopy and zeta-potential shift, and served as a proof-of-concept. Understanding EV corona formation will improve rational EV-inspired nano-therapy design.

摘要

纳米颗粒可以获得定义其生物学特性的血浆蛋白冠。此前，人们认为细胞衍生的细胞外囊泡 (EVs) 的蛋白冠具有功能。在这里，我们证明了在允许的技术条件下，经过规模化切向流过滤 (TFF) 和串联质量标签 (TMT) 蛋白质组学从治疗级人胎盘扩展 (PLX) 基质细胞分泌的纳米大小的 EVs 被可成像和功能化的蛋白冠所包围。Western blot、基于钙黄绿素的流式细胞术、超分辨率和电子显微镜验证了 EV 的身份。PLX-EVs 部分保护了蛋白冠免受蛋白酶的消化。EVs 显著改善了体内的人类皮肤再生和血管生成，诱导了免疫细胞中的差异信号转导，并在体外剂量依赖性地抑制了 T 细胞增殖。通过尺寸排阻或超速离心去除蛋白冠会破坏血管生成。用荧光白蛋白作为模型蛋白或定义的促血管生成因子对 EV 进行伪装以重新建立人工蛋白冠，通过超分辨率显微镜、电子显微镜和 ζ 电位转移进行了描绘，并作为概念验证。了解 EV 蛋白冠的形成将有助于改进基于 EV 的纳米治疗的合理设计。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/93a879746562/JEV2-11-e12207-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/35f08b9ca829/JEV2-11-e12207-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/a971127b16e7/JEV2-11-e12207-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/3de306ac61f9/JEV2-11-e12207-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/08788eda4d09/JEV2-11-e12207-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/aa413bc7e18c/JEV2-11-e12207-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/9c454ef4fa57/JEV2-11-e12207-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/a0b24d271f8f/JEV2-11-e12207-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/93a879746562/JEV2-11-e12207-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/35f08b9ca829/JEV2-11-e12207-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/a971127b16e7/JEV2-11-e12207-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/3de306ac61f9/JEV2-11-e12207-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/08788eda4d09/JEV2-11-e12207-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/aa413bc7e18c/JEV2-11-e12207-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/9c454ef4fa57/JEV2-11-e12207-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/a0b24d271f8f/JEV2-11-e12207-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/998a/8994701/93a879746562/JEV2-11-e12207-g007.jpg

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