Revealing Temozolomide Resistance Mechanisms via Genome-Wide CRISPR Libraries

Autor(es)

Reily Rocha, Clarissa Ribeiro ^[1] ; Rocha, Alexandre Reily ^[2] ; Silva, Matheus Molina ^[3] ; Gomes, Luciana Rodrigues ^[4] ; Latancia, Marcela Teatin ^[3] ; Tomaz, Marina Andrade ^[1] ; de Souza, Izadora ^[1] ; Seregni Monteiro, Linda Karolynne ^[1] ; Martins Menck, Carlos Frederico ^[3]

Afiliação do(s) autor(es)

^[1] Fed Univ Sao Paulo UNIFESP, Dept Clin & Expt Oncol, BR-04037003 Sao Paulo - Brazil

^[2] State Univ Sao Paulo UNESP, Inst Theoret Phys, BR-01140070 Sao Paulo - Brazil

^[3] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, BR-05508000 Sao Paulo - Brazil

^[4] Butantan Inst, Ctr Toxins Immune Response & Cell Signaling CeT, Lab Cell Cycle, BR-05503001 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: CELLS; v. 9, n. 12 DEC 2020.
• Citações Web of Science: 0

Resumo

Glioblastoma is a severe type of brain tumor with a poor prognosis and few therapy options. Temozolomide (TMZ) is one of these options, however, with limited success, and failure is mainly due to tumor resistance. In this work, genome-wide CRISPR-Cas9 lentiviral screen libraries for gene knockout or activation were transduced in the human glioblastoma cell line, aiming to identify genes that modulate TMZ resistance. The sgRNAs enriched in both libraries in surviving cells after TMZ treatment were identified by next-generation sequencing (NGS). Pathway analyses of gene candidates on knockout screening revealed several enriched pathways, including the mismatch repair and the Sonic Hedgehog pathways. Silencing three genes ranked on the top 10 list (MSH2, PTCH2, and CLCA2) confirm cell protection from TMZ-induced death. In addition, a CRISPR activation library revealed that NRF2 and Wnt pathways are involved in TMZ resistance. Consistently, overexpression of FZD6, CTNNB1, or NRF2 genes significantly increased cell survival upon TMZ treatment. Moreover, NRF2 and related genes detected in this screen presented a robust negative correlation with glioblastoma patient survival rates. Finally, several gene candidates from knockout or activation screening are targetable by inhibitors or small molecules, and some of them have already been used in the clinic. (AU)

• Processo FAPESP: 13/08028-1 - CEGH-CEL - Centro de Estudos do Genoma Humano e de Células-Tronco
• Beneficiário: Mayana Zatz
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 19/19435-3 - Papel de danos no DNA e função mitocondrial em envelhecimento vascular, imune e neurológico (DNA MoVINg)
• Beneficiário: Carlos Frederico Martins Menck
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/21745-0 - Explorando mecanismos de resistência a quimioterápicos em células de câncer via bibliotecas CRISPR
• Beneficiário: Clarissa Ribeiro Reily Rocha
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/25016-2 - Explorando o papel de NRF2 e ciclo circadiano como mediadores de resistência a quimioterápicos
• Beneficiário: Clarissa Ribeiro Reily Rocha
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado