A Pilot Single Cell RNA Sequencing Study Suggested that Conditioned Asymmetric Osteogenic Differentiation through mTOR Pathway in Adolescent Idiopathic Scoliosis (AIS)
Invited conference paper presented and published in conference proceedings
CUHK Authors
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AbstractIntroduction: The bone pathological structural changes of osteocyte lacuno-canalicular network (LCN) in adolescent idiopathic scoliosis (AIS) have been reported in our group. The dysfunction of osteoblasts in AIS might result in differential osteogenic differentiation profile that contribute to prognostication for curve progression. However, the specific differences in cell level that associated with the bilateral spinal facet bone tissue are not yet understood. Our transcriptomic study was first to apply based on single cell RNA sequencing of facet joint bone in AIS.
Methods: Facet joint bone tissue was collected perioperatively from the concave and convex side at the curve apex from an AIS male patients aged 16 years. The single-cell suspensions were obtained and loaded according to the instructions provided with the Chromium Single Cell 3′ Library & Gel Bead Kit v2. Library preparation was performed according to the 10x Genomics Chromium platform guidelines, and the resulting libraries were sequenced using the Illumina NovaSeq 6000 System.
Results: A total of 17,714 cells were captured on the concave and convex side facet joint bone tissue through the 10× single cell platform. 40,433 single-cell transcriptomes were integrated into a single data compendium. The two scRNA data were integrated and unsupervised clustering through SERUAT, and 16 clusters were resolved. Through the identification of differentially expressed genes (DEGs) in Cluster 16, the highly expressed genes (ALPL, SATB2, RUNX2, MRC2) in the Cluster16 cell group were consistent with the characteristics of osteoblast transcriptome. Through GO analysis, it was found that the C16 cell group had the biology process of ossification process. The expression level of FZD5 on the concave side was higher than the convex side. Through KEGG enrichment analysis of DEGs, it was found that the mTOR pathway was significantly enriched on the concave side. The protein to protein interaction network of DEGs in concave osteoblasts revealed that LRP6, LEF1, RUNX1, etc. are involved in the mTOR pathway and can interact with FZD5.
Discussion and Conclusion: To our knowledge, this is the first scRNA study to reveal the imbalance mTOR pathway in osteoblast from bilateral apex facet joint bone. In previous findings, the expression level of Wnt/β-catenin is associated with abnormal bone quality. As a downstream pathway, mTOR pathway may contribute to osteogenic differentiation, and the inhibition of mTOR will affect bone tissue mineralization. FAD5 is the key binding gene between WNT and mTOR pathway. Further study on the imbalanced mechanism of FZD5 in bilateral facet joint bone tissue may affect osteogenic differentiation and bone tissue mineralization through mTOR pathway. Thereby our findings could shed light on the development of novel prognostication and treatment for AIS.
Methods: Facet joint bone tissue was collected perioperatively from the concave and convex side at the curve apex from an AIS male patients aged 16 years. The single-cell suspensions were obtained and loaded according to the instructions provided with the Chromium Single Cell 3′ Library & Gel Bead Kit v2. Library preparation was performed according to the 10x Genomics Chromium platform guidelines, and the resulting libraries were sequenced using the Illumina NovaSeq 6000 System.
Results: A total of 17,714 cells were captured on the concave and convex side facet joint bone tissue through the 10× single cell platform. 40,433 single-cell transcriptomes were integrated into a single data compendium. The two scRNA data were integrated and unsupervised clustering through SERUAT, and 16 clusters were resolved. Through the identification of differentially expressed genes (DEGs) in Cluster 16, the highly expressed genes (ALPL, SATB2, RUNX2, MRC2) in the Cluster16 cell group were consistent with the characteristics of osteoblast transcriptome. Through GO analysis, it was found that the C16 cell group had the biology process of ossification process. The expression level of FZD5 on the concave side was higher than the convex side. Through KEGG enrichment analysis of DEGs, it was found that the mTOR pathway was significantly enriched on the concave side. The protein to protein interaction network of DEGs in concave osteoblasts revealed that LRP6, LEF1, RUNX1, etc. are involved in the mTOR pathway and can interact with FZD5.
Discussion and Conclusion: To our knowledge, this is the first scRNA study to reveal the imbalance mTOR pathway in osteoblast from bilateral apex facet joint bone. In previous findings, the expression level of Wnt/β-catenin is associated with abnormal bone quality. As a downstream pathway, mTOR pathway may contribute to osteogenic differentiation, and the inhibition of mTOR will affect bone tissue mineralization. FAD5 is the key binding gene between WNT and mTOR pathway. Further study on the imbalanced mechanism of FZD5 in bilateral facet joint bone tissue may affect osteogenic differentiation and bone tissue mineralization through mTOR pathway. Thereby our findings could shed light on the development of novel prognostication and treatment for AIS.
All Author(s) ListTang G, Chen HX, Yang GK, Lau YA, Lam TP, Zhu ZZ, Qiu Y, Cheng CJ, Lee YW
Name of ConferenceInternational Research Society of Spinal Deformities Scientific Meeting 2024
Start Date of Conference21/06/2024
End Date of Conference23/06/2024
Place of ConferenceHong Kong
Country/Region of ConferenceHong Kong
Year2024
LanguagesEnglish-United Kingdom
