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Original Article | Open Access

Identification of an intronic enhancer regulating RANKL expression in osteocytic cells

Minglu Yan1, Masayuki Tsukasaki2( ), Ryunosuke Muro1, Yutaro Ando1,3, Kazutaka Nakamura1,4, Noriko Komatsu1, Takeshi Nitta1, Tadashi Okamura5, Kazuo Okamoto2, Hiroshi Takayanagi1 ( )
Department of Immunology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo, Tokyo, Japan
Department of Osteoimmunology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo, Tokyo, Japan
Department of Microbiology, Tokyo Dental College, Tokyo, Japan
Department of Oral and Maxillofacial Surgery, Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan
Department of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine, Tokyo, Japan
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Abstract

The bony skeleton is continuously renewed throughout adult life by the bone remodeling process, in which old or damaged bone is removed by osteoclasts via largely unknown mechanisms. Osteocytes regulate bone remodeling by producing the osteoclast differentiation factor RANKL (encoded by the TNFSF11 gene). However, the precise mechanisms underlying RANKL expression in osteocytes are still elusive. Here, we explored the epigenomic landscape of osteocytic cells and identified a hitherto-undescribed osteocytic cell-specific intronic enhancer in the TNFSF11 gene locus. Bioinformatics analyses showed that transcription factors involved in cell death and senescence act on this intronic enhancer region. Single-cell transcriptomic data analysis demonstrated that cell death signaling increased RANKL expression in osteocytic cells. Genetic deletion of the intronic enhancer led to a high-bone-mass phenotype with decreased levels of RANKL in osteocytic cells and osteoclastogenesis in the adult stage, while RANKL expression was not affected in osteoblasts or lymphocytes. These data suggest that osteocytes may utilize a specialized regulatory element to facilitate osteoclast formation at the bone surface to be resorbed by linking signals from cellular senescence/death and RANKL expression.

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Bone Research
Volume 11,
December 2023
Article number: 43
DOI: 10.1038/s41413-023-00277-6
Cite this article:
Yan M, Tsukasaki M, Muro R, et al. Identification of an intronic enhancer regulating RANKL expression in osteocytic cells. Bone Research, 2023, 11: 43. https://doi.org/10.1038/s41413-023-00277-6

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Received: 27 January 2023
Revised: 22 June 2023
Accepted: 05 July 2023
Published: 11 August 2023
© The Author(s) 2023

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