Introduction
Disclosed herein is a formulation comprising an extracellular vesicle (EV), and a therapeutic active agent induced or embedded in the EV. According to preferred embodiments of the present disclosure, the EV is isolated from umbilical cord mesenchymal stem cells, and the active agent may be a growth factor, an immune-modulating agent, a small molecule, an siRNA, cDNA or a plant ingredient (for example, curcumin).
According to certain embodiments, the present formulations are prepared by the method comprising the steps of, (1) isolating the EV from the mesenchymal stem cells; and (2) subjecting the mesenchymal stem cells to a pre-treatment of the active agent; and/or (3) isolating the EVs from (1) or (2) for incorporation of other active agents so as to encapsulate the active agent in the isolated EV.
Also disclosed herein are uses of the present formulation in the treatment of various diseases, including inflammatory diseases, dry eye, degenerative disease, cancer, infectious diseases, rejuvenaion and/or anti-aging in a subject. The formulation disclosed herein may be administered to the subject by a suitable route, including topical, mucosal (e.g. intraconjunctival, intranasal, intratracheal), oral, intraspinal (e.g. intrathecal), intravenous, intraarterial, intramuscular, subcutaneous, intraarticular, intraventrical, intracerebroventricular, intraperitoneal injection and intra-middle ear administration.
Features / strengths
1) EV源自各種細胞優於微脂體或條件細胞培養液
胞外體(extracellular vesicle, EV)是一種源自各種細胞的膜狀囊泡，代表細胞間訊息傳遞的內源機制。一般來說，一細胞(即一供體細胞)可釋放不同形式之EV，包含外泌體(exosomes，直徑介於50到150奈米之間)及微囊泡(microvesicle，直徑介於50到500奈米之間)。一受體細胞會藉由胞飲作用(endocytosis)、膜融合(membrane fusion)或特定的配體-受器內化反應(ligand-receptor internalization)來吸收釋出的EV，據以將EV之內含物(例如，供體細胞的核酸、脂多醣、蛋白及/或脂質)傳遞至受體細胞。
基於EV高生物相容性、低免疫性、奈米級大小及可穿透主要生物屏障(包含血腦障壁)，EV亦可作為一種傳遞藥物的候選療法。
2) EV存有母細胞特性可執行無細胞移植
基於EV源自母細胞特性，具雙層脂質的特性，EV可以佈局作為一種無細胞的移植(Cell-free Transplantation) 的候選療法。如右圖所示以臍帶間質幹細胞(MSCs)為例:
一個幹細胞可以分泌數千個EV，而且可以攜帶細胞間連結溝通的生長素或miRNA，並透過主要生物膜到達諸如腎臟、肝臟及中樞神經系流等遠端器官。
有鑑於此，相關領域亟需一種經修飾的EV，其可廣泛治療不同疾病，例如發炎性疾病、退化性疾病、感染性疾病、代謝性疾病、發育不良性疾病、糖尿病及癌症。
3) 多樣EVs製劑可3代人互補再生精準醫療
利用前條件培養刺激(pre-conditional stimulation)或分離後包覆(post-isolation incorporation)方法，將小分子、胜肽、核酸、適體(aptamer)或骨架(scaffold)包覆或包埋於EV結構中，或許可改善及增加其治療應用性。以臍帶間質幹細胞EV為例，利用上述多樣製備方式，可以提供下列優勢應用:
1. EV具有替代間質幹細胞，傳遞訊息抗發炎和抗退化疾病的治療作用。
2. 天然EV比微脂體更容易進入各種退化器官。
3. EV冷凍或乾燥保存可維持活性和穩定運送。
4. 比母細胞容易製備成皮膚、口服、黏膜和注射製劑。
5. 可開發收集前調理和收集後帶藥的專利佈局。
6. 嬰兒疾病，祖父母退化疾病或父母親炎症疾病的三代互補再生醫學發展。
Specification in detail
