Engineering a Pro-Osteogenic Secretome through the Transient Silencing of the Gene Encoding Secreted Frizzled Related Protein 1.
Engineering a Pro-Osteogenic Secretome through the Transient Silencing of the Gene Encoding Secreted Frizzled Related Protein 1.
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DOI：
10.3390/ijms241512399
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发表时间：
2023-08-03
影响因子：
5.6
通讯作者：
Perez-Campo, Flor Maria
中科院分区：
文献类型：
--
作者：
Garcia-Sanchez, Daniel;Gonzalez-Gonzalez, Alberto;Alvarez-Iglesias, Itziar;del Dujo-Gutierrez, Monica;Bolado-Carrancio, Alfonso;Certo, Matilde;Perez-Nunez, Maria Isabel;Riancho, Jose A.;Rodriguez-Rey, Jose Carlos;Delgado-Calle, Jesus;Perez-Campo, Flor Maria
支持间充质干细胞(MSCs)分泌体再生特性的证据促使了范式的改变，MSCs已从被认为是组织再生的直接贡献者，转变为生产生物技术药物的细胞工厂。我们之前已经设计了一种方法，通过沉默Wnt/β-Catenin抑制剂Sfpr1来促进MSCs向成骨分化。这种方法显著增加了骨质疏松小鼠的骨形成。在这项目前的工作中，我们开始研究SFRP1已经沉默的MSCs的分泌组对体内骨再生的积极作用。小鼠骨髓间充质干细胞系C3H10T1/2的条件培养液(CM-SFRP1)在体外可诱导该细胞系的成骨分化增加，并降低Wnt抑制物Dkk1在小鼠骨细胞中的表达。在体外也检测到RANKL/OPG比率的降低，表明CM-SFRP1对破骨细胞的形成有负面影响。此外，这种CM显著增加了从骨质疏松症患者体外分离的人原代MSCs的矿化。蛋白质组学分析表明，在该分泌体的可溶性和囊泡部分中，参与成骨的蛋白质丰富。总之，我们展示了以这种方式启动的MSCs分泌体的促成骨潜力，表明这是一种有效的方法来增强MSCs分泌体的骨再生特性。
The evidence sustaining the regenerative properties of mesenchymal stem cells’ (MSCs) secretome has prompted a paradigm change, where MSCs have shifted from being considered direct contributors to tissue regeneration toward being seen as cell factories for producing biotech medicines. We have previously designed a method to prime MSCs towards osteogenic differentiation by silencing the Wnt/β-Catenin inhibitor Sfpr1. This approach produces a significant increase in bone formation in osteoporotic mice. In this current work, we set to investigate the contribution of the secretome from the MSCs where Sfrp1 has been silenced, to the positive effect seen on bone regeneration in vivo. The conditioned media (CM) of the murine MSCs line C3H10T1/2, where Sfrp1 has been transiently silenced (CM-Sfrp1), was found to induce, in vitro, an increase in the osteogenic differentiation of this same cell line, as well as a decrease of the expression of the Wnt inhibitor Dkk1 in murine osteocytes ex vivo. A reduction in the RANKL/OPG ratio was also detected ex vivo, suggesting a negative effect of CM-Sfrp1 on osteoclastogenesis. Moreover, this CM significantly increases the mineralization of human primary MSCs isolated from osteoportotic patients in vitro. Proteomic analysis identified enrichment of proteins involved in osteogenesis within the soluble and vesicular fractions of this secretome. Altogether, we demonstrate the pro-osteogenic potential of the secretome of MSCs primmed in this fashion, suggesting that this is a valid approach to enhance the osteo-regenerative properties of MSCs’ secretome.
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影响因子：
14.9
作者：
Swayze EE;Siwkowski AM;Wancewicz EV;Migawa MT;Wyrzykiewicz TK;Hung G;Monia BP;Bennett CF
通讯作者：
Bennett CF
与数据依赖性质谱法结合使用的珠子消化：定量和动态相互作用蛋白质组学的快捷方式。
影响因子：
4.2
作者：
Turriziani B;Garcia-Munoz A;Pilkington R;Raso C;Kolch W;von Kriegsheim A
通讯作者：
von Kriegsheim A
影响因子：
14
作者：
Narayanan K;Kumar S;Padmanabhan P;Gulyas B;Wan ACA;Rajendran VM
通讯作者：
Rajendran VM
影响因子：
4.1
作者：
Delgado-Calle, Jesus;Sanudo, Carolina;Riancho, Jose A.
通讯作者：
Riancho, Jose A.
影响因子：
4
作者：
Inkson, Colette A.;Ono, Mitsuaki;Kuznetsov, Sergei A.;Fisher, Larry W.;Robey, Pamela Gehron;Young, Marian F.
通讯作者：
Young, Marian F.