Receptor activity‐modifying protein 1 regulates the phenotypic expression of BMSCs via the Hippo/Yap pathway

Abstract

Receptor activity‐modifying protein 1 (RAMP1) might be a critical regulator during bone wound healing. However, the roles and mechanisms of RAMP1 in osteogenesis remain unclear. Here, we aimed to elucidate the role of RAMP1 and explore the effects of Yes‐associated protein 1 (Yap1), an effector of the Hippo/Yap pathway, in this process. We used a RAMP1 overexpression lentiviral system in bone marrow mesenchymal stem cells (BMSCs), which enhanced RAMP1 expression in an effective, appropriate, and sustained manner. Alkaline phosphatase (ALP) activity assays and alizarin red staining showed that RAMP1 promoted osteogenic differentiation of BMSCs after calcitonin gene‐related peptide (CGRP) treatment (10 −8 mol/L). Moreover, real‐time polymerase chain reaction and Western blot analysis indicated that RAMP1 upregulated the expression of osteogenic phenotypic markers (ALP, runt‐related transcription factor 2, osteopontin; p < 0.05). To further uncover the mechanism of RAMP1 in osteogenic differentiation, we used verteporfin (10 −7 mol/L) to block Yap1. Notably, verteporfin impaired RAMP1‐induced osteogenesis. Taken together, our findings confirmed that RAMP1 is a key mediator of bone regeneration and indicate that RAMP1 promotes CGRP‐induced osteogenic differentiation of BMSCs via regulation of the Hippo/Yap pathway.

Neuromodulation of bone: Role of different peptides and their interactions (Review)

“…CGRP promotes the proliferation and osteogenic activity of both osteoblasts and their precursor cells in a dose-dependent manner, as observed in primary osteoblastic cells isolated from the calvaria of newborn rabbits, by increasing the levels of intracellular cAMP and upregulating expression of activating transcription factor (45). The Hippo/Yap pathway is also involved in the osteogenic phenotype mediated by the G-protein coupled receptor RAMP1 (46). Coupling with osteogenesis, CGRP exerts an angiogenic effect in the early stage during bone regeneration and ischemia, partly due to its vasodilator capacity and stimulation of endothelial cell migration (11,47).…”

“…CGRP is a nociceptive neurotransmitter released from capsaicin-sensitive sensory nerves in response to various stimulations ( 6 , 13 , 43 ). The CGRP receptor complex contains the calcitonin receptor-like receptor and its co-receptor, receptor activity modifying protein 1 (RAMP1); expression of these receptor complexes have been demonstrated on osteoclasts, osteoblasts and their precursors, and BMMs ( 44 – 46 ). Although the distribution of CGRP positive fibers in skeleton is similar to that of SP ( 7 ), they do not exert similar functions in bone metabolism.…”

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.…”

Fendiline Enhances the Cytotoxic Effects of Therapeutic Agents on PDAC Cells by Inhibiting Tumor-Promoting Signaling Events: A Potential Strategy to Combat PDAC

“…Our published studies showed that fendiline significantly inhibits proliferation, survival and metastatic characteristics of PDAC cells, as evident by reduced migration, invasion and anchorage independent growth [40]. In the current study we tested if combining fendiline with the YAP1 inhibitor visudyne (Verteporfin), which is used for photodynamic therapy of macular degeneration, [41,42,43,44,45,46], elicits additive inhibitory effects on survival and metastatic characteristics of PDAC cells. Additionally, experiments were also conducted with combinations of fendiline and gemcitabine, the nucleoside analog commonly used to treat PDAC.…”