title: Combination of Nanoparticle-Delivered siRNA for Astrocyte Elevated Gene-1 (AEG-1) and All-trans Retinoic Acid (ATRA): An Effective Therapeutic Strategy for Hepatocellular Carcinoma (HCC).
abstract: Hepatocellular carcinoma (HCC) is a fatal cancer with no effective therapy. Astrocyte elevated gene-1 (AEG-1) plays a pivotal role in hepatocarcinogenesis and inhibits retinoic acid-induced gene expression and cell death. The combination of a lentivirus expressing AEG-1 shRNA and all-trans retinoic acid (ATRA) profoundly and synergistically inhibited subcutaneous human HCC xenografts in nude mice. We have now developed liver-targeted nanoplexes by conjugating poly(amidoamine) (PAMAM) dendrimers with polyethylene glycol (PEG) and lactobionic acid (Gal) (PAMAM-PEG-Gal) which were complexed with AEG-1 siRNA (PAMAM-AEG-1si). The polymer conjugate was characterized by (1)H-NMR, MALDI, and mass spectrometry; and optimal nanoplex formulations were characterized for surface charge, size, and morphology. Orthotopic xenografts of human HCC cell QGY-7703 expressing luciferase (QGY-luc) were established in the livers of athymic nude mice and tumor development was monitored by bioluminescence imaging (BLI). Tumor-bearing mice were treated with PAMAM-siCon, PAMAM-siCon+ATRA, PAMAM-AEG-1si, and PAMAM-AEG-1si+ATRA. In the control group the tumor developed aggressively. ATRA showed little effect due to high AEG-1 levels in QGY-luc cells. PAMAM-AEG-1si showed significant reduction in tumor growth, and the combination of PAMAM-AEG-1si+ATRA showed profound and synergistic inhibition so that the tumors were almost undetectable by BLI. A marked decrease in AEG-1 level was observed in tumor samples treated with PAMAM-AEG-1si. The group treated with PAMAM-AEG-1si+ATRA nanoplexes showed increased necrosis, inhibition of proliferation, and increased apoptosis when compared to other groups. Liver is an ideal organ for RNAi therapy and ATRA is an approved anticancer agent. Our exciting observations suggest that the combinatorial approach might be an effective way to combat HCC.
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Devaraja
authors.initials: D
authors.lastname: Rajasekaran
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Srivastava
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Kareem
authors.initials: K
authors.lastname: Ebeid
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Rachel
authors.initials: R
authors.lastname: Gredler
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Maaged
authors.initials: M
authors.lastname: Akiel
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Jariwala
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Chadia L
authors.initials: CL
authors.lastname: Robertson
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Xue-Ning
authors.initials: XN
authors.lastname: Shen
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Ayesha
authors.initials: A
authors.lastname: Siddiq
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Paul B
authors.initials: PB
authors.lastname: Fisher
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Aliasger K
authors.initials: AK
authors.lastname: Salem
authors.affiliation: †Department of Human and Molecular Genetics, §Massey Cancer Center; and ∥VCU Institute of Molecular Medicine (VIMM), Virginia Commonwealth University, Richmond, Virginia 23298, United States.|‡Department of Pharmaceutical Sciences and Experimental Therapeutics, College of Pharmacy, and ⊥Holden Comprehensive Cancer Center, University of Iowa, Iowa City, Iowa 52242, United States.
authors.forename: Devanand
authors.initials: D
authors.lastname: Sarkar
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D015815:Cell Adhesion Molecules / Q000037:antagonists & inhibitors* / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D003131:Combined Modality Therapy; D015316:Genetic Therapy; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D034741:RNA, Small Interfering / Q000235:genetics*; D016601:RNA-Binding Proteins; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D015815:Cell Adhesion Molecules; C483039:MTDH protein, human; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D014212:Tretinoin