Specific and Efficient Regression of Cancers Harboring KRAS Mutation by Targeted RNA Replacement.
Mutations in the KRAS gene, which persistently activate RAS function, are most frequently found in many types of human cancers. Here, we proposed and verified a new approach against cancers harboring the KRAS mutation with high cancer selectivity and efficient anti-cancer effects based on targeted RNA replacement. To this end, trans-splicing ribozymes from Tetrahymena group I intron were developed, which can specifically target and reprogram the mutant KRAS G12V transcript to induce therapeutic gene activity in cells. Adenoviral vectors containing the specific ribozymes with downstream suicide gene were constructed and then infection with the adenoviruses specifically downregulated KRAS G12V expression and killed KRAS G12V-harboring cancer cells additively upon pro-drug treatment, but it did not affect the growth of wild-type KRAS-expressing cells. Minimal liver toxicity was noted when the adenoviruses were administered systemically in vivo. Importantly, intratumoral injection of the adenoviruses with pro-drug treatment specifically and significantly impeded the growth of xenografted tumors harboring KRAS G12V through a trans-splicing reaction with the target RNA. In contrast, xenografted tumors harboring wild-type KRAS were not affected by the adenoviruses. Therefore, RNA replacement with a mutant KRAS-targeting trans-splicing ribozyme is a potentially useful therapeutic strategy to combat tumors harboring KRAS mutation.
Department of Integrated Life Sciences, Research Institute of Advanced Omics, Dankook University, Yongin 16890, Republic of Korea.
Sung Jin
SJ
Kim
Department of Integrated Life Sciences, Research Institute of Advanced Omics, Dankook University, Yongin 16890, Republic of Korea.
Ju Hyun
JH
Kim
Department of Integrated Life Sciences, Research Institute of Advanced Omics, Dankook University, Yongin 16890, Republic of Korea.
Bitna
B
Yang
Department of Pathology and Immune-network Pioneer Research Center, Dong-A University College of Medicine, Busan 49202, Republic of Korea.
Jin-Sook
JS
Jeong
Department of Integrated Life Sciences, Research Institute of Advanced Omics, Dankook University, Yongin 16890, Republic of Korea. Electronic address: swl0208@dankook.ac.kr.
Seong-Wook
SW
Lee
D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004195:Disease Models, Animal; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D009154:Mutation; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology*; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D052416:Targeted Gene Repair; D020040:Trans-Splicing; D023041:Xenograft Model Antitumor Assays
C117307:KRAS protein, human; D016337:RNA, Catalytic; D012313:RNA; D016283:Proto-Oncogene Proteins p21(ras)
KRAS mutation; RNA replacement; adenoviral vector; group I intron; suicide gene therapy; targeted cancer gene therapy; trans-splicing ribozyme