hCTLA4-Gene-Modified Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMMSCs) Maintain POSTN Secretion to Enhance the Migration Capability of Allogeneic hBMMSCs through the Integrin αvβ3/FAK/ERK Signaling Pathway
hCTLA4-Gene-Modified Human Bone Marrow-Derived Mesenchymal Stem Cells (hBMMSCs) Maintain POSTN Secretion to Enhance the Migration Capability of Allogeneic hBMMSCs through the Integrin αvβ3/FAK/ERK Signaling Pathway
复制标题
hCTLA4 基因修饰的人骨髓间充质干细胞 (hBMMSC) 通过整合素 αvβ3/FAK/ERK 信号通路维持 POSTN 分泌以增强同种异体 hBMMSC 的迁移能力
DOI：
10.1155/2020/3608284
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发表时间：
2020-03-24
影响因子：
4.3
通讯作者：
Dai, Fei
中科院分区：
文献类型：
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作者：
Song, Lei;Zhang, Fei;Dai, Fei
细胞毒性T淋巴细胞相关蛋白4（CTLA 4-）修饰的人骨髓间充质干细胞（hBMMSCs）有望成为骨组织工程的种子细胞。然而，其内在机制尚不清楚。在本研究中，我们研究了CTLA 4修饰的hBMMSCs是否通过维持POT分泌参与同种异体hBMMSCs（allo-hBMMSCs）的迁移。从不同的组中分离hBMMSC，命名为hBMMSC和allo-hBMMSC。用阴性对照（NC）、表达CTLA 4、POSTN或CTLA 4加POSTN的shRNA的空腺病毒或重组腺病毒感染的hBMMSC分别命名为NC hBMMSC、CTLA 4修饰的hBMMSC、POSTN修饰的hBMMSC或CTLA 4 + shPOSTN修饰的hBMMSC。然后将它们与PBMC以1：5的比例与2.5 μg/mL植物血凝素（PHA）共培养。收集共培养物上清液以用抗整合素αvβ3 IgG或阴性对照IgG处理allo-hBMMSC作为对照。在此之后，进行ELISA、Transwell测定、伤口愈合测定和蛋白质印迹。我们发现，在CTLA 4和POSTN修饰的hBMMSCs的培养上清液中，POR 4水平高于与PHA处理的PBMC共培养的NC hBMMSCs。经PHA处理的PBMC与经CTLA 4和POSTN修饰的hBMMSCs共培养，其培养上清可增强allo-hBMMSCs的迁移能力，并激活allo-hBMMSCs中整合素αvβ3/FAK/ERK信号通路。此外，这些诱导作用可被POR 4基因敲低而减弱，并且allo-hBMMSCs的迁移能力可被抗整合素αvβ3 IgG阻断。结论：hCTLA 4基因修饰的hBMMSCs通过整合素αvβ3/FAK/ERK信号通路维持POR 4的分泌，增强同种异体hBMMSCs在T细胞免疫活化环境中的迁移能力。
Cytotoxic T-lymphocyte-associated protein 4- (CTLA4-) modified human bone marrow-derived mesenchymal stem cells (hBMMSCs) might be promising seed cells for bone tissue engineering. However, the underlying mechanism is not clear. In the present study, we investigated whether CTLA4-modified hBMMSCs are involved in the migration of allogeneic hBMMSCs (allo-hBMMSCs) by maintaining POSTN secretion. hBMMSCs were isolated from different groups, named hBMMSCs and allo-hBMMSCs. hBMMSCs that were infected with the negative control (NC), empty adenovirus- or recombinant adenovirus-expressing CTLA4, POSTN, or CTLA4 plus the shRNA of POSTN were named NC hBMMSCs, CTLA4-modified hBMMSCs, POSTN-modified hBMMSCs, or CTLA4+shPOSTN-modified hBMMSCs, respectively. They were then cocultured with PBMCs in a 1 : 5 ratio with 2.5 μg/mL phytohemagglutinin (PHA). The coculture supernatant was collected to treat allo-hBMMSCs with anti-integrin αvβ3 IgG, or negative control IgG, as a control. Following this, ELISA, Transwell assays, wound healing assays, and western blotting were performed. We found that the POSTN level was higher in the culture supernatant of CTLA4- and POSTN-modified hBMMSCs than in NC hBMMSCs cocultured with PBMCs treated with PHA. The migration capability of allo-hBMMSCs was enhanced, and the integrin αvβ3/FAK/ERK signaling pathway in allo-hBMMSCs was activated by the culture supernatant of CTLA4- and POSTN-modified hBMMSCs cocultured with PBMCs treated with PHA. Additionally, these induced effects can be weakened by POSTN knockdown, and the migration capability of allo-hBMMSCs was blocked by anti-integrin αvβ3 IgG. In conclusion, hCTLA4-gene-modified hBMMSCs maintain POSTN secretion to enhance the migration capability of allogeneic hBMMSCs through the integrin αvβ3/FAK/ERK signaling pathway in the T cell immune activation environment.