错义突变
生物
突变
突变
清脆的
癌症研究
免疫检查点
结直肠癌
遗传学
损失函数
基因组编辑
计算生物学
癌症
免疫疗法
基因
表型
作者
Matthew A. Coelho,Emre Karakoc,Shriram G. Bhosle,Emanuel João Gonçalves,Thomas Burgold,Sarah Cooper,Chiara Cattaneo,Vivien Veninga,S. M. Consonni,Cansu Dinçer,Rochel M. Lago,Freddy Gibson,Syd Barthorpe,Claire Hardy,Joel Rein,Mark G. Thomas,Emile E. Voest,Andrew R. Bassett,Mathew J. Garnett
标识
DOI:10.1101/2022.03.29.486051
摘要
Abstract IFNγsignalling underpins host responses to infection, inflammation and anti-tumour immunity. Mutations in the IFNγsignalling pathway cause immunological disorders, haematological malignancies, and resistance to immune checkpoint blockade (ICB) in cancer, however the function of most clinically observed variants remain unknown. Here, we systematically investigate the genetic determinants of IFNγresponse in colorectal cancer cells using CRISPR-Cas9 screens and base editing mutagenesis. Deep mutagenesis of JAK1 with cytidine and adenine base editors, combined with pathway-wide screens, reveal loss-of-function and gain-of-function mutations with clinical precedence, including causal variants in haematological malignancies and mutations detected in patients refractory to ICB. We functionally validate variants of uncertain significance in primary tumour organoids, where engineering missense mutations in JAK1 enhanced or reduced sensitivity to autologous tumour-reactive T cells. By classifying > 300 missense variants altering IFNγ pathway activity, we demonstrate the utility of base editing for mutagenesis at scale, and generate a resource to inform genetic diagnosis.
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