A human organoid drug screen identifies α2-adrenergic receptor signaling as a therapeutic target for cartilage regeneration

  • Cell Stem Cell. 2024 Sep 27:S1934-5909(24)00315-1. doi: 10.1016/j.stem.2024.09.001.
Xiaocui Wei  1 ,  Jingyang Qiu  2 ,  Ruijun Lai  3 ,  Tiantian Wei  2 ,  Zhijie Lin  2 ,  Shijiang Huang  2 ,  Yuanjun Jiang  2 ,  Zhanpeng Kuang  4 ,  Hao Zeng  2 ,  Yan Gong  2 ,  Xiaoling Xie  2 ,  Jun Yang  2 ,  Yue Zhang  2 ,  Sheng Zhang  2 ,  Zhipeng Zou  2 ,  Xuefei Gao  5 ,  Xiaochun Bai  6
Affiliations
  • 1. State Key Laboratory of Organ Failure Research, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; Department of Histology and Embryology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China.
  • 2. State Key Laboratory of Organ Failure Research, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China.
  • 3. Academy of Orthopedics, Guangdong Province, Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases, The Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China.
  • 4. Department of Pediatrics, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
  • 5. Department of Physiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; Department of Respiratory and Critical Care Medicine, The Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, Guangdong, China. Electronic address: [email protected].
  • 6. State Key Laboratory of Organ Failure Research, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; Academy of Orthopedics, Guangdong Province, Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases, The Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China. Electronic address: [email protected].
PMID: 39353427 DOI: 10.1016/j.stem.2024.09.001
Abstract

Directed differentiation of stem cells toward chondrogenesis in vitro and in situ to regenerate cartilage suffers from off-target differentiation and hypertrophic tendency. Here, we generated a cartilaginous Organoid system from human expanded pluripotent stem cells (hEPSCs) carrying a COL2A1mCherry and COL10A1eGFP double reporter, enabling real-time monitoring of chondrogenesis and hypertrophy. After screening 2,040 FDA-approved drugs, we found that α-adrenergic receptor (α-AR) antagonists, especially phentolamine, stimulated chondrogenesis but repressed hypertrophy, while α2-AR agonists reduced chondrogenesis and induced hypertrophy. Phentolamine prevented cartilage degeneration in hEPSC cartilaginous Organoid and human cartilage explant models and stimulated microfracture-activated endogenous skeletal stem cells toward hyaline-like cartilage regeneration without fibrotic degeneration in situ. Mechanistically, α2-AR signaling induced hypertrophic degeneration via cyclic guanosine monophosphate (cGMP)-dependent secretory leukocyte protease inhibitor (SLPI) production. SLPI-deleted cartilaginous Organoid was degeneration resistant, facilitating large cartilage defect healing. Ultimately, targeting α2-AR/SLPI was a promising and clinically feasible strategy to regenerate cartilage via promoting chondrogenesis and repressing hypertrophy.

Keywords
cartilage regeneration; drug screening; human expanded pluripotent stem cells; hypertrophic cartilaginous organoid; secretory leukocyte protease inhibitor; α-adrenergic receptor.
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