Design and synthesis of engineered extracellular vesicles and their biomedical applications

doi:10.12211/2096-8280.2023-010

Synthetic Biology Journal ›› 2024, Vol. 5 ›› Issue (1): 154-173.DOI: 10.12211/2096-8280.2023-010

• Invited Review • Previous Articles Next Articles

Design and synthesis of engineered extracellular vesicles and their biomedical applications

LIU Duo¹^,², LIU Peiyuan¹, LI Lianyue¹, WANG Yaxin¹, CUI Yuhui¹, XUE Huimin¹, WANG Hanjie¹

^1.School of Life Sciences，Tianjin University，Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology，Tianjin Key Laboratory of Function and Application of Biological Macromolecules，Tianjin 300072，China
^2.Frontiers Science Center for Synthetic Biology，School of Chemical Engineering and Technology，Tianjin University，Tianjin 300072，China

Received:2023-02-01 Revised:2023-12-27 Online:2024-03-20 Published:2024-02-29
Contact: WANG Hanjie

工程化细胞外囊泡的设计合成与生物医学应用

刘夺¹^,², 刘培源¹, 李连月¹, 王雅欣¹, 崔钰惠¹, 薛慧敏¹, 王汉杰¹

^1.天津大学生命科学学院，天津市微纳生物材料与检疗技术工程中心，天津市生物大分子结构功能与应用重点实验室，天津 300072
^2.天津大学化工学院，合成生物学前沿科学中心，天津 300072

通讯作者: 王汉杰
作者简介:刘夺（1987—），男，博士，副研究员。研究方向为纳米医药合成生物学，从事细菌、酵母设计再造用于医药合成与在体递送等生命健康应用研究。 E-mail：duo_liu0@tju.edu.cn
王汉杰（1984—），男，博士，教授，天津大学生命科学学院副院长。研究方向为纳米生物学、合成生物学和生物医学工程等多学科交叉领域，从事肠道细菌的合成生物学设计再造及生命健康应用研究。 E-mail：wanghj@tju.edu.cn
基金资助:
国家重点研发计划项目(2019YFA0906500);国家自然科学基金优秀青年项目(32122047);国家自然科学基金面上项目(31971300)

Abstract

Abstract:

In recent years, extracellular vesicles have received increasing attention due to their close association with the occurrence and development of diseases. As mechanism underlying the regulation of extracellular vesicles on the development of diseases and other kinds of biological functions has been explored persistently, their utilization as drug carriers has also been tested by scientists for targeted therapy. Extracellular vesicles as drug carriers have several intrinsic advantages compared to artificial carriers, such as higher biocompatibility, lower immunogenicity, better capacity for biofilm fusion, and particular natural homing effect on intracellular communications. However, the biomedical applications of extracellular vesicles also face challenges with their complicated surface modification, poor drug loading capacity and low product yield. Engineered extracellular vesicles refer to the artificial modification of natural extracellular vesicles to particularly fit with target recipient cells or tissues, which can achieve precise delivery of contained functional molecules and support production at a large scale, thus showing a broad prospect for their biomedical applications. Synthetic biology technology can realize the de novo design and reengineering of chassis cells to support the standardized and modular synthesis of extracellular vesicles. This article first reviews the methods and applications of surface modification and functional molecule encapsulation of extracellular vesicles, and then summarizes strategies for their preparation and production, such as extraction, and purification. In the second section, we envision the role of synthetic biology in promoting the customized design and synthesis of engineered extracellular vesicles to further facilitate fine control on attributes, improve efficiency, and expand applications to make them widely used in human health as soon as possible.

Key words: engineered extracellular vesicles, biomedicine, target therapy, chassis, synthetic biology

摘要：

近年来，细胞外囊泡因其与疾病发生发展密切相关而受到越来越广泛的关注。随着细胞外囊泡参与生命功能调控的机制被不断解析，利用细胞外囊泡作为药物载体，用于靶向治疗的工作也相继开展。细胞外囊泡作为药物载体，与人工载体相比，具有高生物相容性、低免疫原性和良好的生物膜融合能力，同时有着参与细胞通信的归巢效应等天然优势。然而，细胞外囊泡的生物医学应用也存在表面修饰复杂、包载能力差、产量较低等问题，导致使用范围严重受限。工程化细胞外囊泡是指对天然细胞外囊泡进行人工改造，使其能够特异性靶向受体细胞或组织，实现所包载功能分子的精准递送，并支撑可放大生产的模式，从而展现出广阔的生物医学应用前景。合成生物学技术的引入，可实现底盘细胞的从头设计再造，支撑细胞外囊泡的标准化、模块化合成。本文首先概括了工程化细胞外囊泡的表面修饰、工程化细胞外囊泡的功能分子包载的方法和应用；其次总结了工程化细胞外囊泡的生产制备，如提升细胞外囊泡产量的工程化策略、细胞外囊泡的放大生产与提取纯化等；最后展望了合成生物学通过改造底盘细胞基因组、人工设计囊泡表面蛋白、调控分子包载的细胞过程等定制化合成细胞外囊泡的前景。合成生物学技术的发展与使用可推动工程化细胞外囊泡的定制化设计与合成，将进一步精细控制其属性、提升其效能、拓展其应用，争取将其早日广泛应用于人类健康事业。

关键词: 细胞外囊泡, 生物医学, 工程化, 靶向治疗, 底盘细胞, 合成生物学

CLC Number:

LIU Duo, LIU Peiyuan, LI Lianyue, WANG Yaxin, CUI Yuhui, XUE Huimin, WANG Hanjie. Design and synthesis of engineered extracellular vesicles and their biomedical applications[J]. Synthetic Biology Journal, 2024, 5(1): 154-173.

刘夺, 刘培源, 李连月, 王雅欣, 崔钰惠, 薛慧敏, 王汉杰. 工程化细胞外囊泡的设计合成与生物医学应用[J]. 合成生物学, 2024, 5(1): 154-173.

Figures/Tables 5

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修饰分子	来源细胞	受体细胞	目标功能	参考文献
RVG肽	HEK293T	脑神经细胞	治疗阿尔兹海默病	[2]
Anti-CD3，Anti-EGFR	Expi293F	T细胞	杀伤乳腺癌细胞	[3]
LLO	细菌	DC细胞	实现抗原呈递	[4]
MERS-CoV的RBD	细菌	免疫细胞	人工抗原	[6]
SPIKE的RBD	细菌	免疫细胞	人工疫苗	[7]
适配体AS1411	小鼠DC细胞	肿瘤细胞	杀伤癌细胞	[8]
c(RGDyK)肽	MSC	脑血管内皮细胞	治疗缺血脑损伤	[9]
动物源配体	杂合膜融合	血管细胞	促血管生成	[10]
动物源配体	杂合膜融合	巨噬细胞	杀伤癌细胞	[11]
c(RGDyK)肽	MSC	脑血管内皮细胞	治疗缺血脑损伤	[9]
α(FRα)	动物细胞	脑实质	脑部递送	[12]
RVG肽	小鼠DC细胞	神经元细胞	治疗神经损伤	[13]
IMTP肽	骨髓MSC	心肌组织	修复心肌	[14]
iRGD肽	动物细胞	肿瘤细胞	杀伤癌细胞	[15]
GE11肽	HEK293T	乳腺癌细胞	杀伤癌细胞	[16]
ICAM1	DC细胞	DC、T细胞	活化免疫功能	[17]
MFGE8	巨噬细胞	巨噬细胞	活化免疫功能	[18]
CIC2	纤维肉瘤细胞	抗原呈递细胞	活化免疫功能	[19]
CAR	CAR-T	肿瘤细胞	杀伤癌细胞	[20]
PD-1	T细胞	肿瘤细胞	阻断免疫逃逸	[21]
异源抗原	细菌	免疫细胞	活化免疫功能	[22]

修饰分子	来源细胞	受体细胞	目标功能	参考文献
RVG肽	HEK293T	脑神经细胞	治疗阿尔兹海默病	[2]
Anti-CD3，Anti-EGFR	Expi293F	T细胞	杀伤乳腺癌细胞	[3]
LLO	细菌	DC细胞	实现抗原呈递	[4]
MERS-CoV的RBD	细菌	免疫细胞	人工抗原	[6]
SPIKE的RBD	细菌	免疫细胞	人工疫苗	[7]
适配体AS1411	小鼠DC细胞	肿瘤细胞	杀伤癌细胞	[8]
c(RGDyK)肽	MSC	脑血管内皮细胞	治疗缺血脑损伤	[9]
动物源配体	杂合膜融合	血管细胞	促血管生成	[10]
动物源配体	杂合膜融合	巨噬细胞	杀伤癌细胞	[11]
c(RGDyK)肽	MSC	脑血管内皮细胞	治疗缺血脑损伤	[9]
α(FRα)	动物细胞	脑实质	脑部递送	[12]
RVG肽	小鼠DC细胞	神经元细胞	治疗神经损伤	[13]
IMTP肽	骨髓MSC	心肌组织	修复心肌	[14]
iRGD肽	动物细胞	肿瘤细胞	杀伤癌细胞	[15]
GE11肽	HEK293T	乳腺癌细胞	杀伤癌细胞	[16]
ICAM1	DC细胞	DC、T细胞	活化免疫功能	[17]
MFGE8	巨噬细胞	巨噬细胞	活化免疫功能	[18]
CIC2	纤维肉瘤细胞	抗原呈递细胞	活化免疫功能	[19]
CAR	CAR-T	肿瘤细胞	杀伤癌细胞	[20]
PD-1	T细胞	肿瘤细胞	阻断免疫逃逸	[21]
异源抗原	细菌	免疫细胞	活化免疫功能	[22]

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Design and synthesis of engineered extracellular vesicles and their biomedical applications

工程化细胞外囊泡的设计合成与生物医学应用

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