CRISPR Knockout of the HuR Gene Causes a Xenograft Lethal Phenotype.
CRISPR Knockout of the HuR Gene Causes a Xenograft Lethal Phenotype.
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DOI：
10.1158/1541-7786.mcr-16-0361
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发表时间：
2017-06
期刊：
影响因子：
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通讯作者：
Brody JR
中科院分区：
文献类型：
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作者：
Lal S;Cheung EC;Zarei M;Preet R;Chand SN;Mambelli-Lisboa NC;Romeo C;Stout MC;Londin E;Goetz A;Lowder CY;Nevler A;Yeo CJ;Campbell PM;Winter JM;Dixon DA;Brody JR
Pancreatic ductal adenocarcinoma (PDA) is the third leading cause of cancer related deaths in the U.S., while colorectal cancer (CRC) is the third most common cancer. The RNA binding protein HuR (ELAVL1), supports a pro-oncogenic network in gastrointestinal (GI) cancer cells through enhanced HuR expression. Using a publically available database, HuR expression levels were determined to be increased in primary PDA and CRC tumor cohorts as compared to normal pancreas and colon tissues, respectively. CRISPR/Cas9 technology was successfully used to delete the HuR gene in both PDA (MIA PaCa-2 and Hs 766T) and CRC (HCT116) cell lines. HuR deficiency has a mild phenotype, in vitro, as HuR-deficient MIA PaCa-2 (MIA.HuR-KO(−/−)) cells had increased apoptosis when compared to isogenic wild-type (MIA.HuR-WT(+/+)) cells. Using this isogenic system, mRNAs were identified that specifically bound to HuR and were required for transforming a 2D culture into 3D (i.e., organoids). Importantly, HuR-deficient MIA PaCa-2 and Hs 766T cells were unable to engraft tumors in vivo compared to control HuR-proficient cells, demonstrating a unique xenograft lethal phenotype. While not as a dramatic phenotype, CRISPR knockout HuR HCT116 colon cancer cells (HCT.HuR-KO(−/−)) showed significantly reduced in vivo tumor growth compared to controls (HCT.HuR-WT(+/+)). Finally, HuR deletion affects KRAS activity and controls a subset of pro-oncogenic genes. The work reported here supports the notion that targeting HuR is a promising therapeutic strategy to treat GI malignancies.
