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| Status |
Public on Dec 16, 2019 |
| Title |
Transcriptome-wide study of the response of human trabecular meshwork cells to the substrate stiffness increase |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Elevated intraocular pressure, a major risk factor of glaucoma, is caused by the abnormal function of trabecular outflow pathways. Human trabecular meshwork (HTM) tissue plays an important role in the outflow pathways. However, the molecular mechanisms that how TM cells respond to the elevated IOP are largely unknown. We cultured primary HTM cells on polyacrylamide gels with tunable stiffness corresponding to Young's moduli ranging from 1.1 to 50 kPa. Then next‐generation RNA sequencing (RNA‐seq) was performed to obtain the transcriptomic profiles of HTM cells. Bioinformatics analysis revealed that genes related to glaucoma including DCN, SPARC, and CTGF, were significantly increased with elevated substrate stiffness, as well as the global alteration of HTM transcriptome. Extracellular matrix (ECM)‐related genes were selectively activated in response to the elevated substrate stiffness, consistent with the known molecular alteration in glaucoma. Human normal and glaucomatous TM tissues were also obtained to perform RNA‐seq experiments and supported the substrate stiffness‐altered transcriptome profiles from the in vitro cell model. The current study profiled the transcriptomic changes in human TM cells upon increasing substrate stiffness. Global change of ECM‐related genes indicates that the in vitro substrate stiffness could greatly affect the biological processes of HTM cells. The in vitro HTM cell model could efficiently capture the main pathogenetic process in glaucoma patients, and provide a powerful method to investigate the underlying molecular mechanisms.
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| Overall design |
Normal HTM cells with two different densities (cultured by 3 and 5 days, respectively) were cultured on collagen-coated polyacrylamide gels of tunable stiffness with Young’s moduli ranging from 1.1 kPa to 50 kPa. TM tissues of glaucoma patients were also obtained. RNA-seq experiments were performed for all the samples.
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| Web link |
https://0-www-ncbi-nlm-nih-gov.brum.beds.ac.uk/pubmed/32115746
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| Contributor(s) |
Zhang Y, Wang J |
| Citation(s) |
32115746 |
| Submission date |
Nov 29, 2018 |
| Last update date |
Mar 06, 2020 |
| Contact name |
Dong Chen |
| Organization name |
ABLife, Inc.
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| Department |
Center for Genome Analysis
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| Street address |
388 GaoXin 2nd Road, East Lake Hi-Tech Development Zone
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| City |
Wuhan |
| State/province |
Hubei |
| ZIP/Postal code |
430075 |
| Country |
China |
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| Platforms (2) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
| GPL20795 |
HiSeq X Ten (Homo sapiens) |
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| Samples (19)
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| Relations |
| BioProject |
PRJNA507545 |
| SRA |
SRP171120 |
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