title: MUC1 Promoter-Driven DTA as a Targeted Therapeutic Strategy against Pancreatic Cancer.
abstract: Mucin1 (MUC1) is overexpressed in pancreatic ductal adenocarcinoma (PDA) and is associated with tumor aggressiveness, suggesting that MUC1 is a promising therapeutic target for promoter-driven diphtheria toxin A (DTA). Endogenous MUC1 transcript levels were analyzed by quantitative PCR (qPCR) in multiple PDA cells (Capan1, HPAFII, Su.86.86, Capan2, Hs766T, MiaPaCa2, and Panc1). Expression levels were correlated with luciferase activity and cell death after transfection with MUC1 promoter-driven luciferase and DTA constructs. MUC1-positive (+) cells had significantly elevated MUC1 mRNA expression compared with MUC1-negative (-) cells. Luciferase activity was significantly higher in MUC1(+) cells when transfected with MUC1 promoter-driven luciferase and MUC1(+) cells underwent enhanced cell death after transfection with a single dose of MUC1 promoter-driven DTA. IFNγ pretreatment enhanced MUC1 expression in MUC1(-) cells and induced sensitivity to MUC1-DTA therapy. Matched primary and metastatic tumor lesions from clinical specimens revealed similar MUC1 IHC labeling patterns, and a tissue microarray of human PDA biopsies revealed increased immunolabeling with a combination of MUC1 and mesothelin (MSLN) antibodies, compared with either antibody alone. Combining MUC1 with MSLN-targeted DTA enhanced drug efficacy in an in vitro model of heterogeneous PDA. These data demonstrate that MUC1 promoter-driven DTA preferentially kills MUC1-expressing PDA cells and drugs that enhance MUC1 expression sensitize PDA cells with low MUC1 expression.
  
  
  CONCLUSIONS
  MUC1 expression in primary and metastatic lesions provides a rationale for the development of a systemic MUC1 promoter-driven DTA therapy that may be further enhanced by combination with other promoter-driven DTA constructs.
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Renee M
authors.initials: RM
authors.lastname: Tholey
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Shruti
authors.initials: S
authors.lastname: Lal
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Masaya
authors.initials: M
authors.lastname: Jimbo
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Burkhart
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Fernando F
authors.initials: FF
authors.lastname: Blanco
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Cozzitorto
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Josh D
authors.initials: JD
authors.lastname: Eisenberg
authors.affiliation: Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Department of Pathology and the David Rubenstein Pancreatic Cancer Research Center, Memorial Sloan Kettering Cancer Center, New York, New York.
authors.forename: Christine A
authors.initials: CA
authors.lastname: Iacobuzio-Donahue
authors.affiliation: Department of Pathology, UT Southwestern Medical Center; Dallas, Texas.
authors.forename: Agnieszka K
authors.initials: AK
authors.lastname: Witkiewicz
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Melissa
authors.initials: M
authors.lastname: Glbert
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Yeo
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania.
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Brody
authors.affiliation: Lankenau Institute for Medical Research, Wynnewood, Pennsylvania. jordan.winter@jefferson.edu SawickiJ@MLHS.org.
authors.forename: Janet A
authors.initials: JA
authors.lastname: Sawicki
authors.affiliation: Department of Surgery and the Jefferson Pancreas, Biliary, and Related Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania. jordan.winter@jefferson.edu SawickiJ@MLHS.org.
authors.forename: Jordan M
authors.initials: JM
authors.lastname: Winter
mesh_terms: D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D016923:Cell Death; D045744:Cell Line, Tumor; D004167:Diphtheria Toxin / Q000235:genetics / Q000494:pharmacology*; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000494:pharmacology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D000090204:Mesothelin; D058990:Molecular Targeted Therapy / Q000379:methods*; D018396:Mucin-1 / Q000235:genetics* / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D010446:Peptide Fragments / Q000235:genetics / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D004167:Diphtheria Toxin; D058851:GPI-Linked Proteins; C000715527:MSLN protein, human; C060192:MUC1 protein, human; D018396:Mucin-1; D010446:Peptide Fragments; D011994:Recombinant Proteins; C029795:diphtheria toxin fragment A; D007371:Interferon-gamma; D000090204:Mesothelin