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)

• Published in: Bioconjugate chemistry Vol. 26; no. 8; pp. 1651 - 1661
• Main Authors: Rajasekaran, Devaraja, Srivastava, Jyoti, Ebeid, Kareem, Gredler, Rachel, Akiel, Maaged, Jariwala, Nidhi, Robertson, Chadia L, Shen, Xue-Ning, Siddiq, Ayesha, Fisher, Paul B, Salem, Aliasger K, Sarkar, Devanand
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 19.08.2015
• Subjects: Animals; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Cell Adhesion Molecules - antagonists & inhibitors; Cell Adhesion Molecules - genetics; Cell Proliferation - drug effects; Combined Modality Therapy; Gene expression; Genetic Therapy; Humans; Lentivirus; Liver - drug effects; Liver - metabolism; Liver cancer; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Liver Neoplasms - therapy; Mice; Mice, Inbred C57BL; Mice, Nude; Nanoparticles; Nanoparticles - administration & dosage; Nanoparticles - chemistry; Retroviridae; RNA, Small Interfering - genetics; Rodents; Tretinoin - pharmacology; Tumor Cells, Cultured; Tumors; Xenograft Model Antitumor Assays
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/acs.bioconjchem.5b00254

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 1H-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.