title: CRISPR Knockout of the HuR Gene Causes a Xenograft Lethal Phenotype.
abstract: Pancreatic ductal adenocarcinoma (PDA) is the third leading cause of cancer-related deaths in the United States, whereas colorectal cancer 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 colorectal cancer tumor cohorts as compared with 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 colorectal cancer (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 with 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 two-dimensional culture into three dimensional (i.e., organoids). Importantly, HuR-deficient MIA PaCa-2 and Hs 766T cells were unable to engraft tumorsin vivocompared with control HuR-proficient cells, demonstrating a unique xenograft lethal phenotype. Although not as a dramatic phenotype, CRISPR knockout HuR HCT116 colon cancer cells (HCT.HuR-KO(-/-)) showed significantly reducedin vivotumor growth compared with controls (HCT.HuR-WT(+/+)). Finally, HuR deletion affects KRAS activity and controls a subset of pro-oncogenic genes.Implications:The work reported here supports the notion that targeting HuR is a promising therapeutic strategy to treat GI malignancies.Mol Cancer Res; 15(6); 696-707. ©2017 AACR.
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Shruti
authors.initials: S
authors.lastname: Lal
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Edwin C
authors.initials: EC
authors.lastname: Cheung
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Zarei
authors.affiliation: Department of Cancer Biology and University of Kansas Cancer Center, University of Kansas Medical Center, Kansas City, Kansas.
authors.forename: Ranjan
authors.initials: R
authors.lastname: Preet
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Saswati N
authors.initials: SN
authors.lastname: Chand
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Mambelli-Lisboa
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Carmella
authors.initials: C
authors.lastname: Romeo
authors.affiliation: Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, Pennsylvania.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Stout
authors.affiliation: Computational Medicine Center, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Eric
authors.initials: E
authors.lastname: Londin
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Austin
authors.initials: A
authors.lastname: Goetz
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Cinthya Y
authors.initials: CY
authors.lastname: Lowder
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Avinoam
authors.initials: A
authors.lastname: Nevler
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Yeo
authors.affiliation: Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, Pennsylvania.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Campbell
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania. jonathan.brody@jefferson.edu ddixon3@kumc.edu jordan.winter@jefferson.edu.
authors.forename: Jordan M
authors.initials: JM
authors.lastname: Winter
authors.affiliation: Department of Cancer Biology and University of Kansas Cancer Center, University of Kansas Medical Center, Kansas City, Kansas. jonathan.brody@jefferson.edu ddixon3@kumc.edu jordan.winter@jefferson.edu.
authors.forename: Dan A
authors.initials: DA
authors.lastname: Dixon
authors.affiliation: Division of Surgical Research, Department of Surgery, Jefferson Pancreas, Biliary and Related Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania. jonathan.brody@jefferson.edu ddixon3@kumc.edu jordan.winter@jefferson.edu.
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Brody
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D000818:Animals; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003110:Colonic Neoplasms / Q000235:genetics* / Q000473:pathology; D000067780:ELAV-Like Protein 1 / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D055786:Gene Knockout Techniques; D006801:Humans; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology; D010641:Phenotype; D023041:Xenograft Model Antitumor Assays
chemical_list: D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human