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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Revealing Temozolomide Resistance Mechanisms via Genome-Wide CRISPR Libraries

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Author(s):
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]
Total Authors: 9
Affiliation:
[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
Total Affiliations: 4
Document type: Journal article
Source: CELLS; v. 9, n. 12 DEC 2020.
Web of Science Citations: 0
Abstract

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)

FAPESP's process: 13/08028-1 - CEGH-CEL - Human Genome and Stem Cell Research Center
Grantee:Mayana Zatz
Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
FAPESP's process: 19/19435-3 - The role of DNA damage and mitochondrial function in vascular, immune and neurological ageing (DNA MoVINg)
Grantee:Carlos Frederico Martins Menck
Support type: Research Projects - Thematic Grants
FAPESP's process: 19/21745-0 - Exploring drug resistance mechanisms in cancer cells via CRISPR libraries
Grantee:Clarissa Ribeiro Reily Rocha
Support type: Regular Research Grants
FAPESP's process: 15/25016-2 - Exploring the role of NRF2 and circadian cycle as mediators of tumor resistance to chemotherapy
Grantee:Clarissa Ribeiro Reily Rocha
Support type: Scholarships in Brazil - Post-Doctorate