Human umbilical cord blood-mesenchymal stem cell derived exosomes as an efficient nanocarrier for Docetaxel and miR-125a: Formulation optimization and anti-metastatic behaviour

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2023-06-01 DOI:10.1016/j.lfs.2023.121621

Moumita Basak , Biswajit Sahoo , Dharmendra Kumar Chaudhary , SaiBhargav Narisepalli , Swasti Tiwari , Deepak Chitkara , Anupama Mittal

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引用次数: 3

Abstract

Aim

Exosomes, as a nanocarrier for the co-delivery of biologicals and small anticancer molecules is yet in its infancy. Herein, we investigated hUCBMSC derived exosomes as a biogenic nanocarrier for the co-delivery of tumor suppressor miR-125a and microtubule destabilizing Docetaxel (DTX) to target the proliferative and migratory aggressiveness of the murine TNBC 4T1 cells.

Main methods

In this study, hUCBMSCs from the human umbilical cord blood cells (hUCB) were successfully transfected with miR-125a. Thereafter, DTX was encapsulated into both non-transfected and transfected exosomes by optimized mild sonication-incubation technique. The anticancer efficiency of hUCBMSC Exo-DTX and miR-125a Exo-DTX was compared by MTT and morphometric assay. The prominent anti-metastatic behaviour of the latter was confirmed by in-vitro wound healing and transwell invasion assay. Further, the synergistic effect of miR-125a and DTX was confirmed by F-actin and nuclear degradation by confocal and FESEM assay.

Key findings

hUCBMSC exosomes exhibited DTX payload of 8.86 ± 1.97 ng DTX/ μg exosomes and miRNA retention capacity equivalent to 12.31 ± 5.73 %. The co-loaded formulation (miR-125a Exo-DTX) exhibited IC₅₀ at 192.8 ng/ml in 4T1 cells, which is almost 2.36 folds' lower than the free DTX IC₅₀ (472.8 ng/ml). Additionally, miR-125a Exo-DTX treatment caused wound broadening upto 6.14 $\pm$ 0.38 % while treatment with free DTX and miR-125a exosomes alone caused 18.71 $\pm$ 4.5 % and 77.36 $\pm$ 10.4 % of wound closure respectively in 36 h. miR-125a Exo-DTX treatment further exhibited significantly reduced invasiveness of 4T1 cells (by 3.5 ± 1.8 %) along with prominent cytoskeletal degradation and nuclear deformation as compared to the miR-125a exosomes treated group. The miR-125a expressing DTX loaded exosomal formulation clearly demonstrated the synergistic apoptotic and anti-migratory efficiency of the miR-125a Exo-DTX.

Significance

The synergistic anticancer and anti-metastatic effect of miR-125a Exo-DTX was observed due to presence of both DTX and miR-125a as the cargo of hUCBMSC derived exosomes.

Abstract Image

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人脐血间充质干细胞衍生的外泌体作为多西他赛和miR-125a的高效纳米载体:配方优化和抗转移行为

外泌体作为一种用于生物制剂和抗癌小分子共同递送的纳米载体，尚处于起步阶段。在此，我们研究了hUCBMSC衍生的外泌体作为生物纳米载体，用于共递送肿瘤抑制因子miR-125a和微管不稳定的多西他赛（DTX），以靶向小鼠TNBC 4T1细胞的增殖和迁移攻击性。主要方法在本研究中，成功地用miR-125a转染了来自人脐带血细胞（hUCB）的hUCBMSCs。此后，通过优化的温和超声孵育技术将DTX包封到未转染和转染的外泌体中。通过MTT法和形态计量法比较hUCBMSC-Exo-DTX和miR-125a-Exo-DTX的抗癌效率。通过体外伤口愈合和transwell侵袭试验证实了后者显著的抗转移行为。此外，miR-125a和DTX的协同作用通过F-肌动蛋白和共聚焦和FESEM分析的核降解得到证实。关键发现hUCBMSC外泌体的DTX有效载荷为8.86±1.97ngDTX/μg外泌体，miRNA保留能力相当于12.31±5.73%。共载制剂（miR-125a-Exo-DTX）在4T1细胞中的IC50为192.8 ng/ml，几乎是游离DTX IC50（472.8 ng/ml）的2.36倍。此外，miR-125a-Exo-DTX治疗导致伤口加宽达6.14±0.38%，而单独使用游离DTX和miR-125a外泌体治疗在36小时内分别导致18.71±4.5%和77.36±10.4%的伤口闭合。与miR-125a外泌体处理组相比，miR-125a Exo-DTX处理进一步显示出4T1细胞的侵袭性显著降低（3.5±1.8%），同时细胞骨架降解和细胞核变形显著。表达miR-125a的DTX负载的外泌体制剂清楚地证明了miR-125a Exo-DTX的协同凋亡和抗迁移效率。显著性由于DTX和miR-125a作为hUCBMSC衍生的外泌体内的货物存在，观察到miR-125a Exo-DTX具有协同抗癌和抗转移作用。

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来源期刊

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.