Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
Broad Institute of Harvard and MIT, Cambridge, MA, USA.
Nat Chem Biol. 2019 May;15(5):529-539. doi: 10.1038/s41589-019-0263-0. Epub 2019 Apr 15.
Understanding the mechanism of small molecules is a critical challenge in chemical biology and drug discovery. Medicinal chemistry is essential for elucidating drug mechanism, enabling variation of small molecule structure to gain structure-activity relationships (SARs). However, the development of complementary approaches that systematically vary target protein structure could provide equally informative SARs for investigating drug mechanism and protein function. Here we explore the ability of CRISPR-Cas9 mutagenesis to profile the interactions between lysine-specific histone demethylase 1 (LSD1) and chemical inhibitors in the context of acute myeloid leukemia (AML). Through this approach, termed CRISPR-suppressor scanning, we elucidate drug mechanism of action by showing that LSD1 enzyme activity is not required for AML survival and that LSD1 inhibitors instead function by disrupting interactions between LSD1 and the transcription factor GFI1B on chromatin. Our studies clarify how LSD1 inhibitors mechanistically operate in AML and demonstrate how CRISPR-suppressor scanning can uncover novel aspects of target biology.
理解小分子的机制是化学生物学和药物发现中的一个关键挑战。药物化学对于阐明药物机制至关重要，它使小分子结构的变化能够获得结构-活性关系（SAR）。然而，开发系统地改变靶蛋白结构的互补方法，可以为研究药物机制和蛋白质功能提供同样有信息的 SAR。在这里，我们探索了 CRISPR-Cas9 诱变在急性髓细胞白血病（AML）背景下分析赖氨酸特异性组蛋白去甲基酶 1（LSD1）与化学抑制剂相互作用的能力。通过这种称为 CRISPR-抑制子扫描的方法，我们通过表明 LSD1 酶活性对于 AML 存活不是必需的，并且 LSD1 抑制剂通过破坏 LSD1 与染色质上转录因子 GFI1B 之间的相互作用来发挥作用，阐明了药物作用机制。我们的研究阐明了 LSD1 抑制剂在 AML 中如何发挥作用，并展示了 CRISPR-抑制子扫描如何揭示靶标生物学的新方面。
