Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling

Background: Gnathodiaphyseal dysplasia (GDD) is a rare autosomal dominant genetic disease characterized by osteosclerosis of the tubular bones and cemento-osseous lesions of the mandibles. Anoctamin 5 (ANO5) is the pathogenic gene, however, the specific molecular mechanism of GDD remains unclear. He...

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Main Authors: Hongyu Li, Shengnan Wang, Shuai Zhang, Rui Dong, Congcong Miao, Zhenchuan Tian, Ying Hu
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Bone Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352187225000026
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author Hongyu Li
Shengnan Wang
Shuai Zhang
Rui Dong
Congcong Miao
Zhenchuan Tian
Ying Hu
author_facet Hongyu Li
Shengnan Wang
Shuai Zhang
Rui Dong
Congcong Miao
Zhenchuan Tian
Ying Hu
author_sort Hongyu Li
collection DOAJ
description Background: Gnathodiaphyseal dysplasia (GDD) is a rare autosomal dominant genetic disease characterized by osteosclerosis of the tubular bones and cemento-osseous lesions of the mandibles. Anoctamin 5 (ANO5) is the pathogenic gene, however, the specific molecular mechanism of GDD remains unclear. Herein, a knockin (Ano5KI/KI) mouse model expressing the human mutation p.Cys360Tyr was used to investigate the role of Akt signaling in enhanced osteogenesis and decreased osteoclastogenesis in GDD. Methods: Bone marrow-derived macrophages (BMMs) and mouse calvarial osteoblasts (mCOBs) were isolated from homozygous Ano5KI/KI mice and treated with SC79, a specific Akt activator. The differentiation and F-actin ring formation of osteoclasts were examined by TRAP and phalloidin staining, respectively. Osteoblast differentiation and mineralization were examined by ALP and alizarin red staining. The expression of bone remodeling-related factors was measured by qRT-PCR. Results: Akt activation promoted the generation of TRAP-positive multinucleated osteoclasts and the formation of actin rings in Ano5KI/KI BMMs cultures, accompanied by increased expression of Nfatc1, Trap, Dc-stamp, Mmp9, Ctsk, and Atp6v0d2. Additionally, Ano5Cys360Tyr mutation down-regulated the Akt phosphorylation level in osteoblast. ALP activity and matrix mineralization capacity in Ano5KI/KI osteoblast cultures were inhibited after SC79 stimulation, with reduced expression of Runx2, Opn, Col1a1, and Ocn. Conclusion: Akt activation by SC79 stimulation can obviously rescue abnormal increased osteogenesis and decreased osteoclastogenesis in Ano5KI/KI mouse model, which demonstrated that disturbed Akt signaling pathway may play a pivotal role in the pathogenesis of GDD, and an Akt activator is probable a therapeutic target for GDD.
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spelling doaj-art-49936fe5f98145f2b83faa0176d2c9822025-08-20T02:52:23ZengElsevierBone Reports2352-18722025-03-012410182510.1016/j.bonr.2025.101825Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signalingHongyu Li0Shengnan Wang1Shuai Zhang2Rui Dong3Congcong Miao4Zhenchuan Tian5Ying Hu6Beijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBeijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBeijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBeijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBeijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBeijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaCorresponding author at: Beijing Stomatological Hospital, Capital Medical University, No 4 Tiantan West, Dongcheng District, Beijing 100050, China.; Beijing Institute of Dental Research, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, ChinaBackground: Gnathodiaphyseal dysplasia (GDD) is a rare autosomal dominant genetic disease characterized by osteosclerosis of the tubular bones and cemento-osseous lesions of the mandibles. Anoctamin 5 (ANO5) is the pathogenic gene, however, the specific molecular mechanism of GDD remains unclear. Herein, a knockin (Ano5KI/KI) mouse model expressing the human mutation p.Cys360Tyr was used to investigate the role of Akt signaling in enhanced osteogenesis and decreased osteoclastogenesis in GDD. Methods: Bone marrow-derived macrophages (BMMs) and mouse calvarial osteoblasts (mCOBs) were isolated from homozygous Ano5KI/KI mice and treated with SC79, a specific Akt activator. The differentiation and F-actin ring formation of osteoclasts were examined by TRAP and phalloidin staining, respectively. Osteoblast differentiation and mineralization were examined by ALP and alizarin red staining. The expression of bone remodeling-related factors was measured by qRT-PCR. Results: Akt activation promoted the generation of TRAP-positive multinucleated osteoclasts and the formation of actin rings in Ano5KI/KI BMMs cultures, accompanied by increased expression of Nfatc1, Trap, Dc-stamp, Mmp9, Ctsk, and Atp6v0d2. Additionally, Ano5Cys360Tyr mutation down-regulated the Akt phosphorylation level in osteoblast. ALP activity and matrix mineralization capacity in Ano5KI/KI osteoblast cultures were inhibited after SC79 stimulation, with reduced expression of Runx2, Opn, Col1a1, and Ocn. Conclusion: Akt activation by SC79 stimulation can obviously rescue abnormal increased osteogenesis and decreased osteoclastogenesis in Ano5KI/KI mouse model, which demonstrated that disturbed Akt signaling pathway may play a pivotal role in the pathogenesis of GDD, and an Akt activator is probable a therapeutic target for GDD.http://www.sciencedirect.com/science/article/pii/S2352187225000026Ano5GDDAktOsteoblastOsteoclast
spellingShingle Hongyu Li
Shengnan Wang
Shuai Zhang
Rui Dong
Congcong Miao
Zhenchuan Tian
Ying Hu
Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
Bone Reports
Ano5
GDD
Akt
Osteoblast
Osteoclast
title Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
title_full Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
title_fullStr Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
title_full_unstemmed Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
title_short Ano5Cys360Tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing Akt signaling
title_sort ano5cys360tyr mutation leads to bone dysfunction of gnathodiaphyseal dysplasia via disturbing akt signaling
topic Ano5
GDD
Akt
Osteoblast
Osteoclast
url http://www.sciencedirect.com/science/article/pii/S2352187225000026
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