EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis

Idiopathic scoliosis (IS) is the most common form of spinal deformity with unclear pathogenesis. In this study, we first reanalyzed the loci associated with IS, drawing upon previous studies. Subsequently, we mapped these loci to candidate genes using either location-based or function-based strategi...

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Main Authors: Lianlei Wang, Xinyu Yang, Sen Zhao, Pengfei Zheng, Wen Wen, Kexin Xu, Xi Cheng, Qing Li, Anas M Khanshour, Yoshinao Koike, Junjun Liu, Xin Fan, Nao Otomo, Zefu Chen, Yaqi Li, Lulu Li, Haibo Xie, Panpan Zhu, Xiaoxin Li, Yuchen Niu, Shengru Wang, Sen Liu, Suomao Yuan, Chikashi Terao, Ziquan Li, Shaoke Chen, Xiuli Zhao, Pengfei Liu, Jennifer E Posey, Zhihong Wu, Guixing Qiu, DISCO study group (Deciphering Disorders Involving Scoliosis & COmorbidities), Shiro Ikegawa, James R Lupski, Jonathan J Rios, Carol A Wise, Jianguo T Zhang, Chengtian Zhao, Nan Wu
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Language:English
Published: eLife Sciences Publications Ltd 2025-07-01
Series:eLife
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Online Access:https://elifesciences.org/articles/95324
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author Lianlei Wang
Xinyu Yang
Sen Zhao
Pengfei Zheng
Wen Wen
Kexin Xu
Xi Cheng
Qing Li
Anas M Khanshour
Yoshinao Koike
Junjun Liu
Xin Fan
Nao Otomo
Zefu Chen
Yaqi Li
Lulu Li
Haibo Xie
Panpan Zhu
Xiaoxin Li
Yuchen Niu
Shengru Wang
Sen Liu
Suomao Yuan
Chikashi Terao
Ziquan Li
Shaoke Chen
Xiuli Zhao
Pengfei Liu
Jennifer E Posey
Zhihong Wu
Guixing Qiu
DISCO study group (Deciphering Disorders Involving Scoliosis & COmorbidities)
Shiro Ikegawa
James R Lupski
Jonathan J Rios
Carol A Wise
Jianguo T Zhang
Chengtian Zhao
Nan Wu
author_facet Lianlei Wang
Xinyu Yang
Sen Zhao
Pengfei Zheng
Wen Wen
Kexin Xu
Xi Cheng
Qing Li
Anas M Khanshour
Yoshinao Koike
Junjun Liu
Xin Fan
Nao Otomo
Zefu Chen
Yaqi Li
Lulu Li
Haibo Xie
Panpan Zhu
Xiaoxin Li
Yuchen Niu
Shengru Wang
Sen Liu
Suomao Yuan
Chikashi Terao
Ziquan Li
Shaoke Chen
Xiuli Zhao
Pengfei Liu
Jennifer E Posey
Zhihong Wu
Guixing Qiu
DISCO study group (Deciphering Disorders Involving Scoliosis & COmorbidities)
Shiro Ikegawa
James R Lupski
Jonathan J Rios
Carol A Wise
Jianguo T Zhang
Chengtian Zhao
Nan Wu
author_sort Lianlei Wang
collection DOAJ
description Idiopathic scoliosis (IS) is the most common form of spinal deformity with unclear pathogenesis. In this study, we first reanalyzed the loci associated with IS, drawing upon previous studies. Subsequently, we mapped these loci to candidate genes using either location-based or function-based strategies. To further substantiate our findings, we verified the enrichment of variants within these candidate genes across several large IS cohorts encompassing Chinese, East Asian, and European populations. Consequently, we identified variants in the EPHA4 gene as compelling candidates for IS. To confirm their pathogenicity, we generated zebrafish mutants of epha4a. Remarkably, the zebrafish epha4a mutants exhibited pronounced scoliosis during later stages of development, effectively recapitulating the IS phenotype. We observed that the epha4a mutants displayed defects in left-right coordination during locomotion, which arose from disorganized neural activation in these mutants. Our subsequent experiments indicated that the disruption of the central pattern generator (CPG) network, characterized by abnormal axon guidance of spinal cord interneurons, contributed to the disorganization observed in the mutants. Moreover, when knocked down efnb3b, the ligand for Epha4a, we observed similar CPG defects and disrupted left-right locomotion. These findings suggested that ephrin B3-Epha4 signaling is vital for the proper functioning of CPGs, and defects in this pathway could lead to scoliosis in zebrafish. Furthermore, we identified two cases of IS in NGEF, a downstream molecule in the EPHA4 pathway. Collectively, our data provide compelling evidence that neural patterning impairments and disruptions in CPGs may underlie the pathogenesis of IS.
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spelling doaj-art-36d6a46dd9ad46dfb15a67a32eb1b2b82025-08-20T02:39:19ZengeLife Sciences Publications LtdeLife2050-084X2025-07-011310.7554/eLife.95324EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosisLianlei Wang0Xinyu Yang1https://orcid.org/0009-0002-6706-9857Sen Zhao2Pengfei Zheng3https://orcid.org/0000-0002-8729-0850Wen Wen4https://orcid.org/0000-0003-4858-8037Kexin Xu5Xi Cheng6Qing Li7Anas M Khanshour8Yoshinao Koike9https://orcid.org/0000-0002-5431-5572Junjun Liu10Xin Fan11Nao Otomo12Zefu Chen13Yaqi Li14Lulu Li15Haibo Xie16Panpan Zhu17Xiaoxin Li18Yuchen Niu19Shengru Wang20Sen Liu21Suomao Yuan22Chikashi Terao23https://orcid.org/0000-0002-6452-4095Ziquan Li24Shaoke Chen25Xiuli Zhao26https://orcid.org/0000-0001-8238-6311Pengfei Liu27Jennifer E Posey28Zhihong Wu29Guixing Qiu30DISCO study group (Deciphering Disorders Involving Scoliosis & COmorbidities)Shiro Ikegawa31James R Lupski32Jonathan J Rios33https://orcid.org/0000-0002-0969-2184Carol A Wise34https://orcid.org/0000-0002-6790-2194Jianguo T Zhang35Chengtian Zhao36https://orcid.org/0000-0003-1236-914XNan Wu37https://orcid.org/0000-0002-9429-2889Department of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, China; Department of Orthopaedic Surgery, Qilu Hospital of Shandong University, Cheeloo College of Medicine, Shandong University, Jinan, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Department of Orthopaedic Surgery, Qilu Hospital of Shandong University, Cheeloo College of Medicine, Shandong University, Jinan, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United StatesInstitute of Evolution & Marine Biodiversity, College of Marine Life Science, Ocean University of China, Qingdao, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaCenter for Pediatric Bone Biology and Translational Research, Scottish Rite for Children, Dallas, United StatesLaboratory for Statistical and Translational Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanInstitute of Evolution & Marine Biodiversity, College of Marine Life Science, Ocean University of China, Qingdao, ChinaDepartment of Pediatric Endocrine and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning, ChinaLaboratory for Statistical and Translational Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan; Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Newborn Screening Center, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care Hospital, Beijing, ChinaInstitute of Evolution & Marine Biodiversity, College of Marine Life Science, Ocean University of China, Qingdao, ChinaInstitute of Evolution & Marine Biodiversity, College of Marine Life Science, Ocean University of China, Qingdao, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Department of Central Laboratory, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Department of Central Laboratory, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopaedic Surgery, Qilu Hospital of Shandong University, Cheeloo College of Medicine, Shandong University, Jinan, ChinaLaboratory for Statistical and Translational Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Pediatric Endocrine and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning, ChinaDepartment of Medical Genetics, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Baylor Genetics, Houston, United StatesBaylor Genetics, Houston, United StatesDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, China; Department of Central Laboratory, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, China; Department of Central Laboratory, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, ChinaLaboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Yokohama, JapanDepartments of Pediatrics, Texas Children’s Hospital and Baylor College of Medicine, Houston, United States; Texas Children’s Hospital, Houston, United States; Human Genome Sequencing Center, Baylor College of Medicine, Houston, United StatesCenter for Pediatric Bone Biology and Translational Research, Scottish Rite for Children, Dallas, United States; Department of Orthopaedics, University of Texas Southwestern Medical Center, Dallas, United States; McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, United StatesCenter for Pediatric Bone Biology and Translational Research, Scottish Rite for Children, Dallas, United States; Department of Orthopaedics, University of Texas Southwestern Medical Center, Dallas, United States; McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaInstitute of Evolution & Marine Biodiversity, College of Marine Life Science, Ocean University of China, Qingdao, ChinaDepartment of Orthopedic Surgery, State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory of Big Data Innovation and Application for Skeletal Health Medical Care, Beijing, China; Key Laboratory of Big Data for Spinal Deformities, Chinese Academy of Medical Sciences, Beijing, ChinaIdiopathic scoliosis (IS) is the most common form of spinal deformity with unclear pathogenesis. In this study, we first reanalyzed the loci associated with IS, drawing upon previous studies. Subsequently, we mapped these loci to candidate genes using either location-based or function-based strategies. To further substantiate our findings, we verified the enrichment of variants within these candidate genes across several large IS cohorts encompassing Chinese, East Asian, and European populations. Consequently, we identified variants in the EPHA4 gene as compelling candidates for IS. To confirm their pathogenicity, we generated zebrafish mutants of epha4a. Remarkably, the zebrafish epha4a mutants exhibited pronounced scoliosis during later stages of development, effectively recapitulating the IS phenotype. We observed that the epha4a mutants displayed defects in left-right coordination during locomotion, which arose from disorganized neural activation in these mutants. Our subsequent experiments indicated that the disruption of the central pattern generator (CPG) network, characterized by abnormal axon guidance of spinal cord interneurons, contributed to the disorganization observed in the mutants. Moreover, when knocked down efnb3b, the ligand for Epha4a, we observed similar CPG defects and disrupted left-right locomotion. These findings suggested that ephrin B3-Epha4 signaling is vital for the proper functioning of CPGs, and defects in this pathway could lead to scoliosis in zebrafish. Furthermore, we identified two cases of IS in NGEF, a downstream molecule in the EPHA4 pathway. Collectively, our data provide compelling evidence that neural patterning impairments and disruptions in CPGs may underlie the pathogenesis of IS.https://elifesciences.org/articles/95324idiopathic scoliosispathogenesisEPHA4central pattern generators
spellingShingle Lianlei Wang
Xinyu Yang
Sen Zhao
Pengfei Zheng
Wen Wen
Kexin Xu
Xi Cheng
Qing Li
Anas M Khanshour
Yoshinao Koike
Junjun Liu
Xin Fan
Nao Otomo
Zefu Chen
Yaqi Li
Lulu Li
Haibo Xie
Panpan Zhu
Xiaoxin Li
Yuchen Niu
Shengru Wang
Sen Liu
Suomao Yuan
Chikashi Terao
Ziquan Li
Shaoke Chen
Xiuli Zhao
Pengfei Liu
Jennifer E Posey
Zhihong Wu
Guixing Qiu
DISCO study group (Deciphering Disorders Involving Scoliosis & COmorbidities)
Shiro Ikegawa
James R Lupski
Jonathan J Rios
Carol A Wise
Jianguo T Zhang
Chengtian Zhao
Nan Wu
EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
eLife
idiopathic scoliosis
pathogenesis
EPHA4
central pattern generators
title EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
title_full EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
title_fullStr EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
title_full_unstemmed EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
title_short EPHA4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
title_sort epha4 signaling dysregulation links abnormal locomotion and the development of idiopathic scoliosis
topic idiopathic scoliosis
pathogenesis
EPHA4
central pattern generators
url https://elifesciences.org/articles/95324
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