GNAS knockdown suppresses osteogenic differentiation of mesenchymal stem cells via activation of Hippo signaling pathway

Abstract

Bone marrow-derived mesenchymal stem cells (BMSCs) are a suitable option for cellbased tissue engineering therapies due to their ability to renew and differentiate into multiple different tissue types, such as bone. Over the last decade, the effect of GNAS on the regulation of osteoblast differentiation has attracted great attention. Herein, this study aimed to explore the role of GNAS in osteogenic differentiation of MSCs. A total of 85 GNAS f/f male mice were selected for animal experiments and 10 GNAS f/f male mice for BMSC isolation to conduct cell experiments. The mice and BMSCs were treated with Verteporfin (a Hippo signaling pathway inhibitor) to inhibit the Hippo signaling pathway or recombinant adenovirus-expressing Cre to knockout the GNAS expression. Next, computed tomography scan, Von Kossa staining, and alizarin red staining were performed to detect osteogenic differentiation ability. Moreover, immunohistochemistry and alkaline phosphatase (ALP) staining were used to assess the expression of Oc and Osx in femur tissues and ALP activity. At last, the expression of GNAS, osteogenic markers, and factors related to the Hippo signaling pathway was evaluated. Initially, the results displayed successful knockout of the GNAS gene from mice and BMSCs. Moreover, the data indicated that GNAS knockout inhibits expression of Oc, Osx, ALP, BMP-2, and Runx2, and ALP activity. Additionally, GNAS knockout promotes activation of the Hippo signaling pathway, so as to repress osteogenic differentiation. Collectively, depleted GNAS exerts an inhibitory role in osteogenic differentiation of MSCs by activating Hippo signaling pathway, providing a candidate mediator for osteoporosis.

Toxocara canis Infection Alters lncRNA and mRNA Expression Profiles of Dog Bone Marrow

“…Crem knock-out mice can display slightly decreased osteoclast number and increased long bone mass (Liu et al, 2007 ). Guanine nucleotide binding protein subunit α ( gnas ) is a key component of the cell membrane receptor pathway, and downregulation of gnas can repress osteogenic differentiation of bone marrow-derived MSCs, aggravating the progression of osteoporosis (An et al, 2019 ; Zheng X. et al, 2020 ). The expression of itga11 and crem was upregulated 158.10 times and 6.23 times in bone marrow at 36 dpi, respectively, while the expression of gnas was downregulated 86.31 times at 36 dpi, suggesting that although T. canis worms mainly reside in the intestine at 36 dpi, they can still influence the bone marrow, especially the host MSCs.…”

Role of Hippo-YAP Signaling in Osseointegration by Regulating Osteogenesis, Angiogenesis, and Osteoimmunology

“…There is strong evidence indicates that BMSCs are regulated by some biological signals on differentiation into bone tissue via Hippo-YAP pathway. The knockout of GNAS activates Hippo signaling pathway and eventually suppresses osteogenic differentiation of BMSCs (An et al, 2019), consistently, through the regulation of Hippo-YAP signaling pathway RAMP1 can promote the osteogenesis process of BMSCs that induced by CGRP (Zhang et al, 2019). Tang and Weiss (2017) revealed an interaction of Snail/Slug and YAP/TAZ, which regulates the differentiation ability of BMSCs in a cooperative way, suggesting a potential impact on the properties of osteogenesis.…”

Role of Hippo-YAP1/TAZ pathway and its crosstalk in cardiac biology

“…The Hippo-YAP1/TAZ pathway is regulated by different heterotrimeric G proteins. YAP1/TAZ is generally activated by Gα12/13, Gα q/11 , and Gα i ; these G proteins are coupled with different GPCRs, including lysophosphatidic acid receptors, sphingosine1-phosphate (S1P) receptors, and protease-activated receptor 1 [73][74][75][76][77][78][79]. However, Gα s inhibits the Hippo-YAP1/TAZ pathway by other GPCRs, such as free fatty acid receptor 1/4 (FFAR1/4, also called as G-protein receptor 40/120 [GPR40/120]) [80][81][82].…”