Dataset Information

2-Guanidino-quinazoline promotes efficient translational suppression of nonsense mutations responsible of Human genetic diseases

ABSTRACT: Premature translation termination (PTC) codons account for 10%-20% of genetic diseases in humans. To counteract the gene inactivation by a PTC, it is possible to use drugs to stimulate PTC readthrough, restoring the expression of the full-length protein. To broaden the medical applications of this therapeutic strategy it is important to increase the chemical variety of readthrough-inducers. In the present study we have developed a new reporter cell-line and performed a High Throughput Screening (HTS). Using a workflow of three successive assays we have isolated the 2-Guanidino-quinazoline (TLN468) as a potent readthrough inducer. We tested the clinical potential of the drug onto the 40 most frequent PTC responsible for Duchenne muscular dystrophy and show that TLN468 is more efficient and acts on a broader range of sequences than gentamicin without inducing readthrough of natural stop codons.

ORGANISM(S): Homo sapiens

PROVIDER: GSE185985 | GEO | 2022/09/15

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Premature termination codons (PTC) cause over 10% of genetic disease cases. Some aminoglycosides that bind to the ribosome decoding centre can induce PTC readthrough and restore low levels of full-length functional proteins. However, concomitant inhibition of protein synthesis limits the extent of PTC readthrough that can be achieved by aminoglycosides like G418. Using a cell-based screen, we identified a small molecule, the phenylpyrazoleanilide Y-320, that potently enhances TP53, DMD and COL17A1 PTC readthrough by G418. Unexpectedly, Y-320 increased cellular protein levels and protein synthesis, measured by SYPRO Ruby protein staining and puromycin labelling, as well as ribosome biogenesis measured using antibodies to rRNA and ribosomal protein S6. Y-320 did not increase the rate of translation elongation and it exerted its effects independently of mTOR signaling. At the single cell level, exposure to Y-320 and G418 increased ribosome content and protein synthesis which correlated strongly with PTC readthrough. As a single agent, Y-320 did not affect translation fidelity measured using a luciferase reporter gene but it enhanced misincorporation by G418. RNA-seq data showed that Y-320 upregulated the expression of CXC chemokines CXCL10, CXCL8, CXCL2, CXCL11, CXCL3, CXCL1 and CXCL16. Several of these chemokines exert their cellular effects through the receptor CXCR2 and the CXCR2 antagonist SB225002 reduced cellular protein levels and PTC readthrough in cells exposed to Y-320 and G418. These data show that the self-limiting nature of PTC readthrough by G418 can be compensated by Y-320, a potent enhancer of PTC readthrough that increases ribosome biogenesis and protein synthesis. They also support a model whereby increased PTC readthrough is enabled by increased protein synthesis mediated by an autocrine chemokine signaling pathway. The findings also raise the possibility that inflammatory processes affect cellular propensity to readthrough agents and that immunomodulatory drugs like Y-320 might find application in PTC readthrough therapy.

Dataset Information

2-Guanidino-quinazoline promotes efficient translational suppression of nonsense mutations responsible of Human genetic diseases

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets