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| Status |
Public on Jul 20, 2018 |
| Title |
In vitro gene signature of cultured volar or nonvolar keratinocytes |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
How cell and tissue identity persist despite constant cell turnover is an important biologic question with cell therapy implications. While many mechanisms exist, we investigated the controls for site-specific gene expression in skin given its diverse structures and functions. For example, the transcriptome of in vivo palmoplantar (i.e. volar) epidermis is globally unique including Keratin 9 (KRT9). While volar fibroblasts have the capacity to induce KRT9 in non-volar keratinocytes, we demonstrate here that volar keratinocytes continue to express KRT9 in vitro solo-cultures. Despite this, KRT9 expression is lost with volar keratinocyte passaging, in spite of stable hypo-methylation of its promoter. Coincident with KRT9 loss is a gain of the primitive Keratin 7 and a signature of dsRNA sensing, including the dsRNA receptor DDX58. Exogenous dsRNA inhibits KRT9 expression in early passage volar keratinocytes or in vivo footpads of wild-type mice. Loss of DDX58 in passaged volar keratinocytes rescues KRT9 and inhibits KRT7 expression. Additionally, DDX58 null mice are resistant to the ability of dsRNA to inhibit KRT9 expression. These results demonstrate that the sensing of dsRNA is critical for loss of cell specific gene expression; our results have important implications of how dsRNA sensing is important outside of immune pathways.
Keratinocytes were expanded from both the sole and the dorsum of the foot and at passage 4, RNA was extracted and sent for microarray analysis
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| Overall design |
The same single human donor was the source of a single isolate of volar keratinocytes and a single isolate of nonvolar keratinocytes. At passage 4 total RNA was extracted and sent for microarray analysis
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| Contributor(s) |
Garza L |
| Citation(s) |
30120933 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 AR064297 |
Mechanism and clinical pilot tst of volar fibroblast to create ectopic volar skin |
JOHNS HOPKINS UNIVERSITY |
Luis Andres Garza |
| R01 AR068280 |
Towards the clinical testing of dsRNA to promote wound induced hair neogenesis |
JOHNS HOPKINS UNIVERSITY |
Luis Andres Garza |
|
| Submission date |
Jun 20, 2018 |
| Last update date |
Oct 19, 2018 |
| Contact name |
Luis Andres Garza |
| E-mail(s) |
LAG@jhmi.edu
|
| Phone |
410-955-8662
|
| Organization name |
Johns Hopkins School of Medicine
|
| Department |
Dermatology
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| Lab |
Garza lab
|
| Street address |
1550 Orleans St. Suite 204 CRBII Koch
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| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21287 |
| Country |
USA |
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| Platforms (1) |
| GPL15207 |
[PrimeView] Affymetrix Human Gene Expression Array |
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| Samples (2) |
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| This SubSeries is part of SuperSeries: |
| GSE116055 |
In vitro gene signature of keratinocytes |
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| Relations |
| BioProject |
PRJNA476919 |
