Epigenetic reprogramming enhances the therapeutic efficacy of osteoblast-derived extracellular vesicles to promote human bone marrow stem cell osteogenic differentiation.
Epigenetic reprogramming enhances the therapeutic efficacy of osteoblast-derived extracellular vesicles to promote human bone marrow stem cell osteogenic differentiation.
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DOI：
10.1002/jev2.12118
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发表时间：
2021-07
影响因子：
16
通讯作者：
Cox SC
中科院分区：
文献类型：
--
作者：
Man K;Brunet MY;Fernandez-Rhodes M;Williams S;Heaney LM;Gethings LA;Federici A;Davies OG;Hoey D;Cox SC
细胞外囊泡（EV）在组织工程中作为有前途的脱细胞工具出现，规避了与基于细胞的治疗相关的许多限制。通过组蛋白去乙酰化酶（HDAC）抑制的表观遗传调节已显示出增加分化能力。因此，本研究旨在研究使用HDAC抑制剂曲古抑菌素A（TSA）增强成骨细胞表观遗传功能的潜力，以增强成骨细胞衍生EV用于骨再生的治疗功效。TSA被发现通过降低HDAC活性和增加组蛋白乙酰化显著改变成骨细胞表观遗传功能。TSA处理还显著增强了成骨细胞碱性磷酸酶活性（1.35倍）、胶原蛋白生成（2.8倍）和成骨培养过程中钙沉积（1.55倍）（P ≤ 0.001）。与未处理的EV相比，来自TSA处理的成骨细胞的EV（TSA-EV）显示出减小的粒度（1 - 05倍）（P > 0.05）、浓度（1.4倍）（P > 0.05）和蛋白质含量（1.16倍）（P ≤ 0.001）。与未处理的EV相比，TSA‐ EV显著增强了人骨髓干细胞（hBMSC）的增殖（1.13倍）和迁移（1.3倍）（P ≤ 0.05）。此外，与未经处理的EV培养的细胞相比，TSA-EV上调hBMSC成骨细胞相关基因和蛋白表达（ALP，Col 1a，BSP 1和OCN）。重要的是，TSA-EV引起hBMSC细胞外基质矿化的时间-剂量依赖性增加。microRNA分析揭示了一组来自TSA-EV的差异表达的microRNA，其与成骨相关。靶点预测表明这些microRNA参与了“内吞作用”和“Wnt信号通路”等调控途径。此外，蛋白质组学分析确定了TSA-EV中参与转录调控的蛋白质的富集。总之，我们的研究结果表明，改变成骨细胞的表观基因组加速了它们的矿化，并促进了分泌型EV的骨诱导效力，部分原因是前成骨microRNA和转录调节蛋白的递送。因此，我们第一次证明了利用表观遗传调控作为一种新的工程方法来增强EV对骨修复的治疗效果的潜力。表观遗传调节促进成骨细胞衍生的EV用于骨修复的成骨潜能。HDAC抑制剂曲古抑菌素A（TSA）通过超乙酰化改变成骨细胞表观遗传功能，增强其矿化能力。源自TSA处理的成骨细胞的EV（TSA-EV）富含促成骨微RNA和转录调节蛋白。TSA‐ EV显著促进hBMSCs成骨分化和矿化。表观遗传重编程提供了一种新的工程方法，以提高EV作为骨再生的无细胞工具的治疗效果。
Extracellular vesicles (EVs) are emerging in tissue engineering as promising acellular tools, circumventing many of the limitations associated with cell‐based therapies. Epigenetic regulation through histone deacetylase (HDAC) inhibition has been shown to increase differentiation capacity. Therefore, this study aimed to investigate the potential of augmenting osteoblast epigenetic functionality using the HDAC inhibitor Trichostatin A (TSA) to enhance the therapeutic efficacy of osteoblast‐derived EVs for bone regeneration. TSA was found to substantially alter osteoblast epigenetic function through reduced HDAC activity and increased histone acetylation. Treatment with TSA also significantly enhanced osteoblast alkaline phosphatase activity (1.35‐fold), collagen production (2.8‐fold) and calcium deposition (1.55‐fold) during osteogenic culture (P ≤ 0.001). EVs derived from TSA‐treated osteoblasts (TSA‐EVs) exhibited reduced particle size (1‐05‐fold) (P > 0.05), concentration (1.4‐fold) (P > 0.05) and protein content (1.16‐fold) (P ≤ 0.001) when compared to untreated EVs. TSA‐EVs significantly enhanced the proliferation (1.13‐fold) and migration (1.3‐fold) of human bone marrow stem cells (hBMSCs) when compared to untreated EVs (P ≤ 0.05). Moreover, TSA‐EVs upregulated hBMSCs osteoblast‐related gene and protein expression (ALP, Col1a, BSP1 and OCN) when compared to cells cultured with untreated EVs. Importantly, TSA‐EVs elicited a time‐dose dependent increase in hBMSCs extracellular matrix mineralisation. MicroRNA profiling revealed a set of differentially expressed microRNAs from TSA‐EVs, which were osteogenic‐related. Target prediction demonstrated these microRNAs were involved in regulating pathways such as ‘endocytosis’ and ‘Wnt signalling pathway’. Moreover, proteomics analysis identified the enrichment of proteins involved in transcriptional regulation within TSA‐EVs. Taken together, our findings suggest that altering osteoblasts’ epigenome accelerates their mineralisation and promotes the osteoinductive potency of secreted EVs partly due to the delivery of pro‐osteogenic microRNAs and transcriptional regulating proteins. As such, for the first time we demonstrate the potential to harness epigenetic regulation as a novel engineering approach to enhance EVs therapeutic efficacy for bone repair. Epigenetic regulation promotes the osteogenic potency of osteoblast‐derived EVs for bone repair. The HDAC inhibitor Trichostatin A (TSA) altered osteoblast epigenetic functionality through hyperacetylation, enhancing its mineralisation capacity. EVs derived from TSA treated osteoblasts (TSA‐EVs) were enriched with pro‐osteogenic microRNAs and transcriptional regulating proteins. TSA‐EVs significantly promoted hBMSCs osteogenic differentiation and mineralisation. Epigenetic reprogramming provides a novel engineering approach to enhance EVs therapeutic efficacy as an acellular tool for bone regeneration.
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影响因子：
4.1
作者：
Choo, Min-Kyung;Yeo, Hyeonju;Zayzafoon, Majd
通讯作者：
Zayzafoon, Majd
HDAC抑制剂诱导肿瘤 - 细胞选择性促凋亡转录反应。
影响因子：
9
作者：
通讯作者：
--
影响因子：
4.8
作者：
Dudakovic, Amel;Evans, Jared M.;Westendorf, Jennifer J.
通讯作者：
Westendorf, Jennifer J.
DOI：
10.1016/j.biocel.2012.12.008
发表时间：
2013-03-01
影响因子：
4
作者：
Bhushan, Raghu;Gruenhagen, Johannes;Knaus, Petra
通讯作者：
Knaus, Petra
影响因子：
120.1
作者：
EL Andaloussi, Samir;Maeger, Imre;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.