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| Status |
Public on Sep 30, 2024 |
| Title |
A human organoid drug screen identifies α2-adrenergic receptor signaling as a therapeutic target for cartilage regeneration [bulk RNA-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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.
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| Overall design |
To analyze the overall transcriptome features of cultured cells and dissect subpopulations of hypertrophic cartilaginous organoids, we used FCM-sorted cells to conduct RNA sequencing (RNA-seq) on COL2+COL10- and COL2+COL10+ cell populations generated from day 42 organoids.
We next performed RNA sequencing of D42 PM-treated and vehicle-treated pellets to comprehensively understand the effects of targeting a-AR on chondrogenic phenotypes.
To identify downstream targets through which SLPI promoted hypertrophic differentiation, we conducted RNA-seq of day 42 SLPI KO and control pellets.
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| Contributor(s) |
Bai X |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
Dec 03, 2023 |
| Last update date |
Sep 30, 2024 |
| Contact name |
zhijie Lin |
| E-mail(s) |
linzhijie1943@outlook.com
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| Organization name |
Southern Medical University
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| Street address |
Baiyun Jingxi road
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| City |
Guangzhou |
| ZIP/Postal code |
510000 |
| Country |
China |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (18)
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| This SubSeries is part of SuperSeries: |
| GSE271815 |
A human organoid drug screen identifies α2-adrenergic receptor signaling as a therapeutic target for cartilage regeneration |
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| Relations |
| BioProject |
PRJNA1048064 |
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