Oncogenic mutations in the KRAS gene are well-established drivers of cancer. While the recently developed KRASG12C inhibitors offer a targeted KRAS therapy and have shown success in the clinic, KRASG12C represents only 11% of all KRAS mutations. Current therapeutic approaches for all other KRAS mutations are both indirect and non-mutant-selective, largely focusing on inhibition of downstream KRAS effectors such as MAP kinases. Inhibition of KRAS downstream signaling results in a system-wide down-modulation of the respective targets, raising concerns about systemic cell toxicity. Here, we describe a custom short interfering RNA (siRNA) oligonucleotide (EFTX-D1) designed to preferentially bind mRNA of the most commonly occurring KRAS missense mutations in codons 12 and 13. We determined that EFTX-D1 preferentially reduced the mutant KRAS sequence versus wild-type at the levels of both transcription and translation, and reversed oncogenic KRAS-induced morphologic and growth transformation. Furthermore, EFTX-D1 significantly impaired the proliferation of several KRAS mutant cancer cell lines in 2-D as well as 3-D assays. Taken together, our data indicate a novel use of RNA interference (RNAi) to target oncogenic KRAS-driven cancers specifically.

中文翻译：

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突变选择性小干扰RNA沉默致癌性KRAS

KRAS基因中的致癌突变是公认的癌症驱动因素。尽管最近开发的KRASG12C抑制剂提供了靶向性KRAS治疗并已在临床上显示出成功，但KRASG12C仅代表所有KRAS突变的11％。当前所有其他KRAS突变的治疗方法都是间接和非突变选择性的，主要集中在抑制下游KRAS效应子（如MAP激酶）上。抑制KRAS下游信号传导导致各个靶标在系统范围内的下调，引起对全身细胞毒性的担忧。在这里，我们描述了一种定制的短干扰RNA（siRNA）寡核苷酸（EFTX-D1），旨在优先结合密码子12和13中最常见的KRAS错义突变的mRNA。我们确定EFTX-D1在转录和翻译的水平上均优先降低了突变KRAS序列相对于野生型，并逆转了致癌性KRAS诱导的形态和生长转化。此外，EFTX-D1在2-D和3-D分析中显着损害了几种KRAS突变癌细胞系的增殖。两者合计，我们的数据表明RNA干扰（RNAi）的新型用途专门针对致癌性KRAS驱动的癌症。