Abstract

Comprehensive screenings to clarify indirect cell–cell interactions, such as those in the tumor microenvironment, especially comprehensive assessments of supporting cells’ effects, are challenging. Therefore, in this study, indirect CRISPR screening for drug resistance with cell–cell interactions was invented. The photoconvertible fluorescent protein Dendra2 was inducted to supporting cells and explored the drug resistance responsible factors of supporting cells with CRISPR screenings. Random mutated supporting cells co-cultured with leukemic cells induced drug resistance with cell–cell interactions. Supporting cells responsible for drug resistance were isolated with green-to-red photoconversion, and 39 candidate genes were identified. Knocking out C9orf89, MAGI2, MLPH, or RHBDD2 in supporting cells reduced the ratio of apoptosis of cancer cells. In addition, the low expression of RHBDD2 in supporting cells, specifically fibroblasts, of clinical pancreatic cancer showed a shortened prognosis, and a negative correlation with CXCL12 was observed. Indirect CRISPR screening was established to isolate the responsible elements of cell–cell interactions. This screening method could reveal unknown mechanisms in all kinds of cell–cell interactions by revealing live phenotype-inducible cells, and it could be a platform for discovering new targets of drugs for conventional chemotherapies.

An indirect CRISPR screening system based on the photoconvertible fluorescent protein, Dendra2, enables the discovery of elements responsible for drug resistance from cell–cell interactions.

Details

Title
Indirect CRISPR screening with photoconversion revealed key factors of drug resistance with cell–cell interactions
Author
Sugita, Keisuke 1   VIAFID ORCID Logo  ; Onishi, Iichiroh 1 ; Nakayama, Ran 1 ; Ishibashi, Sachiko 1 ; Ikeda, Masumi 1 ; Inoue, Miori 1 ; Narita, Rina 1 ; Oshima, Shiori 1 ; Shimizu, Kaho 1 ; Saito, Shinichiro 1 ; Sato, Shingo 2   VIAFID ORCID Logo  ; Moriarity, Branden S. 3 ; Yamamoto, Kouhei 1 ; Largaespada, David A. 3   VIAFID ORCID Logo  ; Kitagawa, Masanobu 1 ; Kurata, Morito 1   VIAFID ORCID Logo 

1  Tokyo Medical and Dental University (TMDU), Department of Comprehensive Pathology, Graduate School of Medical and Dental Sciences, Tokyo, Japan (GRID:grid.265073.5) (ISNI:0000 0001 1014 9130) 
2  Tokyo Medical and Dental University (TMDU), Center for Innovative Cancer Treatment, Tokyo, Japan (GRID:grid.265073.5) (ISNI:0000 0001 1014 9130) 
3  University of Minnesota, Masonic Cancer Center, Minneapolis, USA (GRID:grid.17635.36) (ISNI:0000000419368657) 
Pages
582
Publication year
2023
Publication date
2023
Publisher
Nature Publishing Group
e-ISSN
23993642
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s42003-023-04941-9
ProQuest document ID
2821509595
Copyright
© The Author(s) 2023. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.