Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.
Laboratory Medicine Program, Toronto General Hospital, University Health Network, Toronto, ON, Canada.
Blood Adv. 2023 May 23;7(10):2252-2270. doi: 10.1182/bloodadvances.2022008966.
Bit by bit, over the last few decades, functional genomic tools have been piecing together the molecular puzzle driving tumorigenesis in human patients. Nevertheless, our understanding of the role of several genes and regulatory elements that drive critical cancer-associated physiological processes from disease development to progression to spread is very limited, which significantly affects our ability of applying these insights in the context of improved disease management. The recent advent of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based technology and its application in cancer genomics has, however, allowed the generation of a wealth of knowledge that has helped decipher several critical questions associated with translational cancer research. Precisely, the high-throughput capability coupled with a high level of technological plasticity associated with the CRISPR-Cas9 screens have expanded our horizons from a mere struggle to appreciate cancer as a genetic disease to observing the integrated genomic/epigenomic network of numerous malignancies and correlating it with our present knowledge of drugging strategies to develop innovative approaches for next-generation precision cancer medicine. Specifically, within blood cancers, current CRISPR screens have specifically focused on improving our understanding of drug resistance mechanisms, disease biology, the development of novel therapeutic approaches, and identifying the molecular mechanisms of current therapies, with an underlying aim of improving disease outcomes. Here, we review the development of the CRISPR-Cas9 genome-editing strategy, explicitly focusing on the recent advances in the CRISPR-Cas9-based screening approaches, its current capabilities, limitations, and future applications in the context of hematological malignancies.
几十年来,功能基因组学工具逐渐拼凑出导致人类患者肿瘤发生的分子谜题。然而,我们对驱动关键癌症相关生理过程的几个基因和调控元件的作用的理解非常有限,从疾病的发展到进展再到扩散,这极大地影响了我们在改善疾病管理的背景下应用这些见解的能力。最近,基于成簇规律间隔短回文重复(CRISPR)-CRISPR 相关蛋白 9(Cas9)的技术的出现及其在癌症基因组学中的应用,已经产生了大量的知识,有助于解决与转化癌症研究相关的几个关键问题。确切地说,与 CRISPR-Cas9 筛选相关的高通量能力和高水平的技术可塑性,使我们的视野从仅仅将癌症视为一种遗传疾病,扩展到观察众多恶性肿瘤的综合基因组/表观基因组网络,并将其与我们目前对药物治疗策略的了解相关联,以开发用于下一代精准癌症医学的创新方法。具体来说,在血液癌症中,当前的 CRISPR 筛选特别侧重于提高我们对耐药机制、疾病生物学、新治疗方法的开发以及当前疗法的分子机制的理解,其根本目的是改善疾病结果。在这里,我们回顾了 CRISPR-Cas9 基因组编辑策略的发展,特别关注基于 CRISPR-Cas9 的筛选方法的最新进展、其当前能力、局限性以及在血液恶性肿瘤中的未来应用。