Site-specific mutagenesis screening in KRASG12D mutant library to uncover resistance mechanisms to KRASG12D inhibitors

• Published in: Cancer letters 2024-06, Vol.591, p.216904, Article 216904
• Main Authors: Choi, Jeesoo, Shin, Ju-young, Kim, Taeyul K., Kim, Kiwook, Kim, Jiyun, Jeon, Eunhye, Park, Juyeong, Han, Yoon Dae, Kim, Kyung-A, Sim, Taebo, Kim, Hui Kwon, Kim, Han Sang
• Format: Article
• Language: English
• Subjects: Drug resistance; KRAS inhibitor; Site-specific mutagenesis screening
• ISSN: 0304-3835; 1872-7980; 1872-7980
• DOI: 10.1016/j.canlet.2024.216904

KRAS plays a crucial role in regulating cell survival and proliferation and is one of the most commonly mutated oncogenes in human cancers. The novel KRASG12D inhibitor, MRTX1133, demonstrates promising antitumor efficacy in vitro and in vivo. However, the development of acquired resistance in treated patients presents a considerable challenge to sustained therapeutic effectiveness. In response to this challenge, we conducted site-specific mutagenesis screening to identify potential secondary mutations that could induce resistance to MRTX1133. We screened a range of KRASG12D variants harboring potential secondary mutations, and 44 representative variants were selected for in-depth validation of the pooled screening outcomes. We identified eight variants (G12D with V9E, V9W, V9Q, G13P, T58Y, R68G, Y96W, and Q99L) that exhibited substantial resistance, with V9W showing notable resistance, and downstream signaling analyses and structural modeling were conducted. We observed that secondary mutations in KRASG12D can lead to acquired resistance to MRTX1133 and BI-2865, a novel pan-KRAS inhibitor, in human cancer cell lines. This evidence is critical for devising new strategies to counteract resistance mechanisms and, ultimately, enhance treatment outcomes in patients with KRASG12D-mutant cancers. •Secondary mutation in KRASG12D induces resistance to MRTX1133.•Site-specific mutagenesis screening identifies resistance-conferring variants.•The V9W mutation in KRASG12D shows significant resistance.•Downstream signaling analyses and structural modeling were utilized.•These insights aid in devising strategies against acquired resistance.