Targeting BRAF mutant pre-mRNA alternative splicing in melanoma
Alternative splicing of BRAF mRNA can provide a resistance mechanism to BRAF and MEK inhibitors that has been described mostly in BRAF-mutant melanoma. The mechanisms underlying the production of a variety of BRAF mRNA isoforms, e.g., the BRAF3-9 (which lacks exons 4 to 8), remain poorly understood....
| Autor: | |
|---|---|
| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | CBUC, CESCA |
| Repositorio: | TDR. Tesis Doctorales en Red |
| OAI Identifier: | oai:www.tdx.cat:10803/687762 |
| Acceso en línea: | http://hdl.handle.net/10803/687762 |
| Access Level: | acceso abierto |
| Palabra clave: | Alternative splicing Melanoma BRAF Drug resistance MAPK Splicing alternativo Resistencia a terapias 575 |
| Sumario: | Alternative splicing of BRAF mRNA can provide a resistance mechanism to BRAF and MEK inhibitors that has been described mostly in BRAF-mutant melanoma. The mechanisms underlying the production of a variety of BRAF mRNA isoforms, e.g., the BRAF3-9 (which lacks exons 4 to 8), remain poorly understood. Analysis of RNA-seq from melanoma samples identified for the first time BRAF mRNA isoforms associated with resistance in wild type BRAF and in treatment-naïve melanoma samples. Using minigene assays and whole-genome sequencing, we have reasonably rule out a contribution of single-nucleotide sequence variants (such as intronic mutations) in the generation of these isoforms. Using a CRISPR-Cas9 knockout screen, we have identified genetic vulnerabilities related to splicing and chromosome dynamics in melanoma cells, but not splicing factors particularly involved in the generation of BRAF3-9. Importantly, we have identified large intragenic deletions as the underlying mechanism for the production of BRAF3-9 and BRAF1-9 isoforms, suggesting that this can be a general mechanism for the production of resistance-associated mRNA variants in melanoma. |
|---|