Single-cell transcriptome profiling identifies the activation of type I interferon signaling in ossified posterior longitudinal ligament

Abstract

Ossification of the posterior longitudinal ligament (OPLL) is a condition comprising ectopic bone formation from spinal ligaments. This disease is a leading cause of myelopathy in the Asian population. However, the molecular mechanism underlying OPLL and efficient preventive interventions remain unclear. Here, we performed single-cell RNA sequencing and revealed that type I interferon (IFN) signaling was activated in the ossified ligament of patients with OPLL. We also observed that IFN-β stimulation promoted the osteogenic differentiation of preosteoblasts in vitro and activated the ossification-related gene SPP1, thereby confirming the single-cell RNA sequencing findings. Further, blocking the IFN-α/β subunit 1 receptor (IFNAR1) using an anti-IFNAR1 neutralizing antibody markedly suppressed osteogenic differentiation. Together, these results demonstrated that the type I IFN signaling pathway facilitated ligament ossification, and the blockade of this signaling might provide a foundation for the prevention of OPLL.

Keywords

ossification / posterior longitudinal ligament / osteogenic differentiation / IFN signaling

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Acknowledgements

This work was supported by grants from the Ministry of Science and Technology of the People’s Republic of China (No. 2022YFA1104003 to Hongquan Zhang and No. 2021YFC2501003 to Hongquan Zhang); and the National Natural Science Foundation of China (No. 82073076 to Yu Yu, No. 82372992 to Yu Yu, No. 82230094 to Hongquan Zhang and No. 81972616 to Hongquan Zhang, and No. 82372451 to Xiaoguang Liu).

Compliance with ethics guidelines

Conflicts of interest Xiao Liu, Lei Zhang, Ge Wang, Wei Zhao, Chen Liang, Youzhi Tang, Yenan Fu, Bo Liu, Jing Zhang, Xiaoguang Liu, Hongquan Zhang, and Yu Yu declare that they have no conflict of interest.

The study was approved by the PUTH Institutional Review Board (IRB00006761-M2019410), and the study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. Informed consent was obtained from all patients for being included in the study.

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