Systemic delivery of siRNA by actively targeted polyion complex micelles for silencing the E6 and E7 human papillomavirus oncogenes

Haruka Nishida; Yoko Matsumoto; Kei Kawana; R James Christie; Mitsuru Naito; Beob Soo Kim; Kazuko Toh; Hyun Su Min; Yu Yi; Yu Matsumoto; Hyun Jin Kim; Kanjiro Miyata; Ayumi Taguchi; Kensuke Tomio; Aki Yamashita; Tomoko Inoue; Hiroe Nakamura; Asaha Fujimoto; Masakazu Sato; Mitsuyo Yoshida; Katsuyuki Adachi; Takahide Arimoto; Osamu Wada-Hiraike; Katsutoshi Oda; Takeshi Nagamatsu; Nobuhiro Nishiyama; Kazunori Kataoka; Yutaka Osuga; Tomoyuki Fujii

doi:10.1016/j.jconrel.2016.03.016

Systemic delivery of siRNA by actively targeted polyion complex micelles for silencing the E6 and E7 human papillomavirus oncogenes

J Control Release. 2016 Jun 10:231:29-37. doi: 10.1016/j.jconrel.2016.03.016. Epub 2016 Mar 12.

Authors

Haruka Nishida¹, Yoko Matsumoto¹, Kei Kawana², R James Christie³, Mitsuru Naito³, Beob Soo Kim⁴, Kazuko Toh³, Hyun Su Min⁴, Yu Yi⁴, Yu Matsumoto³, Hyun Jin Kim³, Kanjiro Miyata⁵, Ayumi Taguchi¹, Kensuke Tomio¹, Aki Yamashita¹, Tomoko Inoue¹, Hiroe Nakamura¹, Asaha Fujimoto¹, Masakazu Sato¹, Mitsuyo Yoshida¹, Katsuyuki Adachi¹, Takahide Arimoto¹, Osamu Wada-Hiraike¹, Katsutoshi Oda¹, Takeshi Nagamatsu¹, Nobuhiro Nishiyama⁶, Kazunori Kataoka⁷, Yutaka Osuga¹, Tomoyuki Fujii¹

Affiliations

¹ Department of Obstetrics and Gynecology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
² Department of Obstetrics and Gynecology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan. Electronic address: kkawana-tky@umin.org.
³ Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
⁴ Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
⁵ Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. Electronic address: miyata@bmw.t.u-tokyo.ac.jp.
⁶ Polymer Chemistry Division, Chemical Resources Laboratory, Tokyo Institute of Technology, R1-11, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan; Innovation Center of NanoMedicine, Institute of Industry Promotion-Kawasaki, 3-25-14 Tonomachi, Kawasaki-ku, Kawasaki 210-0821, Japan.
⁷ Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Department of Materials Engineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan; Innovation Center of NanoMedicine, Institute of Industry Promotion-Kawasaki, 3-25-14 Tonomachi, Kawasaki-ku, Kawasaki 210-0821, Japan. Electronic address: kataoka@bmw.t.u-tokyo.ac.jp.

PMID: 26979870
DOI: 10.1016/j.jconrel.2016.03.016

Abstract

Human papillomavirus (HPV) E6 and E7 oncogenes are essential for the immortalization and maintenance of HPV-associated cancer and are ubiquitously expressed in cervical cancer lesions. Small interfering RNA (siRNA) coding for E6 and E7 oncogenes is a promising approach for precise treatment of cervical cancer, yet a delivery system is required for systemic delivery to solid tumors. Here, an actively targeted polyion complex (PIC) micelle was applied to deliver siRNAs coding for HPV E6/E7 to HPV cervical cancer cell tumors in immune-incompetent tumor-bearing mice. A cell viability assay revealed that both HPV type 16 and 18 E6/E7 siRNAs (si16E6/E7 and si18E6/E7, respectively) interfered with proliferation of cervical cancer cell lines in an HPV type-specific manner. A fluorescence imaging biodistribution analysis further revealed that fluorescence dye-labeled siRNA-loaded PIC micelles efficiently accumulated within the tumor mass after systemic administration. Ultimately, intravenous injection of si16E6/E7 and si18E6/E7-loaded PIC micelles was found to significantly suppress the growth of subcutaneous SiHa and HeLa tumors, respectively. The specific activity of siRNA treatment was confirmed by the observation that p53 protein expression was restored in the tumors excised from the mice treated with si16E6/E7- and si18E6/E7-loaded PIC micelles for SiHa and HeLa tumors, respectively. Therefore, the actively targeted PIC micelle incorporating HPV E6/E7-coding siRNAs demonstrated its therapeutic potential against HPV-associated cancer.

Keywords: Cervical cancer; Drug delivery system; Human papillomavirus (HPV) E6/E7; Polyion complex (PIC) micelle; Small interfering RNA (siRNA).

MeSH terms

Animals
Cell Line, Tumor
Cell Proliferation
DNA-Binding Proteins / genetics*
Drug Carriers
Female
Gene Expression
Gene Silencing
Heterografts
Humans
Mice, Inbred BALB C
Mice, Nude
Mice, SCID
Micelles
Oncogene Proteins, Viral / genetics*
Papillomaviridae
Papillomavirus E7 Proteins / genetics*
Polyethylene Glycols / chemistry
Polylysine / chemistry
RNA, Small Interfering / administration & dosage*
Repressor Proteins / genetics*
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism
Uterine Cervical Neoplasms / pathology
Uterine Cervical Neoplasms / therapy
Uterine Cervical Neoplasms / virology

Substances

DNA-Binding Proteins
Drug Carriers
E6 protein, Human papillomavirus type 16
E6 protein, Human papillomavirus type 18
E7 protein, Human papillomavirus type 18
Micelles
Oncogene Proteins, Viral
Papillomavirus E7 Proteins
RNA, Small Interfering
Repressor Proteins
Tumor Suppressor Protein p53
oncogene protein E7, Human papillomavirus type 16
Polylysine
Polyethylene Glycols