Identification of genes functionally involved in the detrimental effects of mutant histone H3.3-K27M in Drosophila melanogaster
| Autor: | Nada Jabado, Nicolas De Jay, Margret Shirinian, Martin Hasselblatt, Paul Lasko, Johannes Berlandi, Astrid Jeibmann, Amel Chaouch, Claudia L. Kleinman, Isabel Tegeder, Katharina Thiel |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Cancer Research
Candidate gene Mutagenesis (molecular biology technique) Biology Chromatin remodeling Histones 03 medical and health sciences Histone H3 0302 clinical medicine Biomarkers Tumor Animals Humans RNA Small Interfering High-Throughput Nucleotide Sequencing Glioma biology.organism_classification Chromatin Cell biology High-Throughput Screening Assays Histone Drosophila melanogaster Oncology 030220 oncology & carcinogenesis Basic and Translational Investigations Mutation biology.protein Neurology (clinical) PRC2 030217 neurology & neurosurgery |
| Zdroj: | Neuro Oncol |
| Popis: | Background Recurrent specific mutations in evolutionarily conserved histone 3 (H3) variants drive pediatric high-grade gliomas (HGGs), but little is known about their downstream effects. The aim of this study was to identify genes involved in the detrimental effects of mutant H3.3-K27M, the main genetic driver in lethal midline HGG, in a transgenic Drosophila model. Methods Mutant and wild-type histone H3.3-expressing flies were generated using a φC31-based integration system. Genetic modifier screens were performed by crossing H3.3-K27M expressing driver strains and 194 fly lines expressing short hairpin RNA targeting genes selected based on their potential role in the detrimental effects of mutant H3. Expression of the human orthologues of genes with functional relevance in the fly model was validated in H3-K27M mutant HGG. Results Ubiquitous and midline glia-specific expression of H3.3-K27M but not wild-type H3.3 caused pupal lethality, morphological alterations, and decreased H3K27me3. Knockdown of 17 candidate genes shifted the lethal phenotype to later stages of development. These included histone modifying and chromatin remodeling genes as well as genes regulating cell differentiation and proliferation. Notably, several of these genes were overexpressed in mutant H3-K27M mutated HGG. Conclusions Rapid screening, identification, and validation of relevant targets in "oncohistone" mediated pathogenesis have proven a challenge and a barrier to providing novel therapies. Our results provide further evidence on the role of chromatin modifiers in the genesis of H3.3-K27M. Notably, they validate Drosophila as a model system for rapid identification of relevant genes functionally involved in the detrimental effects of H3.3-K27M mutagenesis. |
| Databáze: | OpenAIRE |
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