Temporal transcriptome features identify early skeletal commitment during human epiphysis development at single-cell resolution

Zhonghao Deng; Shengwei Rong; Lu Gan; Fuhua Wang; Liangxiao Bao; Fang Cai; Zheting Liao; Yu Jin; Shuhao Feng; Zihang Feng; Yiran Wei; Ruge Chen; Yangchen Jin; Yanli Zhou; Xiaoyong Zheng; Liping Huang; Liang Zhao

doi:10.1016/j.isci.2023.107200

Temporal transcriptome features identify early skeletal commitment during human epiphysis development at single-cell resolution

iScience. 2023 Jun 24;26(8):107200. doi: 10.1016/j.isci.2023.107200. eCollection 2023 Aug 18.

Authors

Zhonghao Deng^{1

2}, Shengwei Rong^{1

2}, Lu Gan^{1

2}, Fuhua Wang^{1

2}, Liangxiao Bao¹, Fang Cai³, Zheting Liao^{1

2}, Yu Jin^{1

2}, Shuhao Feng^{1

2}, Zihang Feng^{1

2}, Yiran Wei^{1

2}, Ruge Chen^{1

2}, Yangchen Jin^{1

2}, Yanli Zhou⁴, Xiaoyong Zheng⁵, Liping Huang⁴, Liang Zhao^{1

2

6}

Affiliations

¹ Department of Orthopaedic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China.
² Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University, Guangzhou, Guangdong 510515, China.
³ Department of Obstetrics and Gynecology, Southern Medical University Nanfang Hospital Taihe Branch, Guangzhou, Guangdong 510515, China.
⁴ Department of Obstetrics and Gynecology, Southern Medical University Nanfang Hospital, Guangzhou, Guangdong 510515, China.
⁵ Orthopaedic Department, The 8th medical center of Chinese PLA General Hospital, Beijing 100091, China.
⁶ Department of Orthopaedic Surgery, Shunde First People Hospital, Foshan, Guangdong 528300, China.

Abstract

Human epiphyseal development has been mainly investigated through radiological and histological approaches, uncovering few details of cellular temporal genetic alternations. Using single-cell RNA sequencing, we investigated the dynamic transcriptome changes during post-conception weeks (PCWs) 15-25 of human distal femoral epiphysis cells. We find epiphyseal cells contain multiple subtypes distinguished by specific markers, gene signatures, Gene Ontology (GO) enrichment analysis, and gene set variation analysis (GSVA). We identify the populations committed to cartilage or ossification at this time, although the secondary ossification centers (SOCs) have not formed. We describe the temporal alternation in transcriptional expression utilizing trajectories, transcriptional regulatory networks, and intercellular communication analyses. Moreover, we find the emergence of the ossification-committed population is correlated with the COL2A1-(ITGA2/11+ITGB1) signaling. NOTCH signaling may contribute to the formation of cartilage canals and ossification via NOTCH signaling. Our findings will advance the understanding of single-cell genetic changes underlying fetal epiphysis development.

Keywords: cell biology; developmental biology; transcriptomics.