| Autor: |
Vardapour, Romina, Kehl, Tim, Kneitz, Susanne, Ludwig, Nicole, Meese, Eckart, Lenhof, Hans-Peter, Gessler, Manfred |
| Předmět: |
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| Zdroj: |
Carcinogenesis; Feb2022, Vol. 43 Issue 2, p82-93, 12p |
| Abstrakt: |
Wilms tumor (WT) is the most common renal tumor in childhood. We and others have previously identified oncogenic driver mutations affecting the microprocessor genes DROSHA and DGCR8 that lead to altered miRNA expression patterns. In the case of DGCR8 , a single recurrent hotspot mutation (E518K) was found in the RNA binding domain. To functionally assess this mutation in vitro , we generated mouse Dgcr8 -KO embryonic stem cell (mESC) lines with an inducible expression of wild-type or mutant DGCR8 , mirroring the hemizygous mutant expression seen in WT. RNA-seq analysis revealed significant differences of miRNA expression profiles in DGCR8-E518K compared with DGCR8-wild-type mESCs. The E518K mutation only led to a partial rescue of the reported miRNA processing defect in Dgcr8 -KO, with selectively reduced expression of numerous canonical miRNAs. Nevertheless, DGCR8-E518K retained significant activity given its ability to still process many miRNAs. Subsequent to altered miRNA levels, the expression of mRNA targets was likewise changed. Functional assays showed that DGCR8-E518K cells still have a partial proliferation and differentiation defect but were able to rescue critical biological processes in embryoid body development. The stem cell program could be shut down and all three germ layers were formed. These findings suggest that the E518K mutation leads to a partial reduction of microprocessor activity and altered specificity with selective impairment only in certain developmental contexts, apparently including nephrogenesis. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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