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CRISPR筛选鉴定出肿瘤3D生长特有弱点
2020-03-12 20:49
来源：小柯机器人

近日，美国斯坦福大学医学院Michael C. Bassik、Kyuho Han等研究人员合作利用肿瘤球状体CRISPR筛选鉴定出肿瘤3D生长中特有的弱点。这一研究成果于2020年3月11日在线发表在《自然》杂志上。

研究人员设计了可扩展的癌症球状体模型，并在2D单层和3D肺癌球状体中进行了全基因组CRISPR筛选。3D中的CRISPR表型更准确地概括了体内肿瘤的表型，并且在2D和3D条件之间敏感性不同的基因高度富集了肺癌中突变的基因。这些分析还揭示了对于3D和体内癌症生长至关重要的驱动程序，而非2D。值得注意的是，研究人员发现羧肽酶D负责从胰岛素样生长因子1受体的α链中去除C端RKRR基序（这对受体活性至关重要）。羧肽酶D的表达与肺癌患者的预后相关，而羧肽酶D的缺失降低了肿瘤的生长。这些研究结果揭示了2D和3D癌症模型之间的关键差异，并建立了在球状体中进行CRISPR筛选来揭示癌症弱点的通用策略。
 
据了解，癌症基因组学研究已经鉴定出数千种可能的癌症驱动基因。开发高通量且精确的模型来定义这些基因的功能是一项重大挑战。
 
附：英文原文

Title: CRISPR screens in cancer spheroids identify 3D growth-specific vulnerabilities

Author: Kyuho Han, Sarah E. Pierce, Amy Li, Kaitlyn Spees, Grace R. Anderson, Jose A. Seoane, Yuan-Hung Lo, Michael Dubreuil, Micah Olivas, Roarke A. Kamber, Michael Wainberg, Kaja Kostyrko, Marcus R. Kelly, Maryam Yousefi, Scott W. Simpkins, David Yao, Keonil Lee, Calvin J. Kuo, Peter K. Jackson, Alejandro Sweet-Cordero, Anshul Kundaje, Andrew J. Gentles, Christina Curtis, Monte M. Winslow, Michael C. Bassik

Issue&Volume: 2020-03-11

Abstract: Cancer genomics studies have identified thousands of putative cancer driver genes1. Development of high-throughput and accurate models to define the functions of these genes is a major challenge. Here we devised a scalable cancer-spheroid model and performed genome-wide CRISPR screens in 2D monolayers and 3D lung-cancer spheroids. CRISPR phenotypes in 3D more accurately recapitulated those of in vivo tumours, and genes with differential sensitivities between 2D and 3D conditions were highly enriched for genes that are mutated in lung cancers. These analyses also revealed drivers that are essential for cancer growth in 3D and in vivo, but not in 2D. Notably, we found that carboxypeptidase D is responsible for removal of a C-terminal RKRR motif2 from the α-chain of the insulin-like growth factor 1 receptor that is critical for receptor activity. Carboxypeptidase D expression correlates with patient outcomes in patients with lung cancer, and loss of carboxypeptidase D reduced tumour growth. Our results reveal key differences between 2D and 3D cancer models, and establish a generalizable strategy for performing CRISPR screens in spheroids to reveal cancer vulnerabilities.

DOI: 10.1038/s41586-020-2099-x

Source: https://www.nature.com/articles/s41586-020-2099-x

Nature：《自然》，创刊于1869年。隶属于施普林格·自然出版集团，最新IF：69.504
官方网址：http://www.nature.com/
投稿链接：http://www.nature.com/authors/submit_manuscript.html


本期文章：《自然》：Online/在线发表

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