Phosphoinositide-focused CRISPR screen identifies novel genetic vulnerabilities in PDAC and AML cells

bioRxiv - Cancer Biology Pub Date : 2024-09-09 DOI:10.1101/2024.09.09.612071

Daniel KC Lee, Ryan Loke, Jonathan TS Chow, Martino Marco Gabra, Leonardo Salmena

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Abstract

Phosphoinositides (PIs) are minor but essential phospholipids that play crucial roles in cellular signaling pathways, membrane dynamics, and the regulation of various cellular processes. We developed and utilized a novel PI-focused CRISPR gRNA library to perform negative-selection and positive-selection screens in PANC-1 and OCI-AML2 cells, models of pancreatic ductal adenocarcinoma (PDAC) and acute myeloid leukemia (AML), respectively. Through these screens, we identified 28 essential genes in PANC-1, 84 essential genes in OCI-AML2, and 28 regulators of colony formation in OCI-AML2. Our results using this small and focused library uncovered false negatives and subtle effects that may be missed in genome-wide approaches, while enabling adaptation to different screening conditions. Overall, our results uncovered previously uncharacterized essential genes in PDAC and AML that can be leveraged as therapeutic targets and biomarkers. We also demonstrate that focused libraries offer a more efficient and targeted approach to uncovering critical genetic determinants of cancer progression.

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以磷酸肌酸为重点的 CRISPR 筛选发现了 PDAC 和 AML 细胞中的新型基因漏洞

磷脂（PI）是一种次要但重要的磷脂，在细胞信号通路、膜动力学和各种细胞过程的调控中发挥着至关重要的作用。我们开发并利用一种新型的以 PI 为重点的 CRISPR gRNA 文库，在 PANC-1 和 OCI-AML2 细胞（分别是胰腺导管腺癌（PDAC）和急性髓性白血病（AML）的模型）中进行了阴性选择和阳性选择筛选。通过这些筛选，我们在 PANC-1 中发现了 28 个重要基因，在 OCI-AML2 中发现了 84 个重要基因，在 OCI-AML2 中发现了 28 个集落形成调节因子。我们利用这个小而集中的文库发现了假阴性和全基因组方法可能忽略的微妙效应，同时还能适应不同的筛选条件。总之，我们的研究结果发现了 PDAC 和 AML 中以前未表征的重要基因，这些基因可被用作治疗靶点和生物标记物。我们还证明，聚焦文库为发现癌症进展的关键基因决定因素提供了一种更高效、更有针对性的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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bioRxiv - Cancer Biology

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