Efficient delivery of miR-122 to regulate cholesterol metabolism using a non-covalent peptide-based strategy

Wang,Lilin; Tang,Wei; Yan,Shirong; Zhou,Long; Shen,Tao; Huang,Xiuqing; Dou,Lin; Wang,Mo; Yu,Songlin; Li,Jian

doi:10.3892/mmr.2013.1691

Efficient delivery of miR-122 to regulate cholesterol metabolism using a non-covalent peptide-based strategy

Authors:
- Lilin Wang
- Wei Tang
- Shirong Yan
- Long Zhou
- Tao Shen
- Xiuqing Huang
- Lin Dou
- Mo Wang
- Songlin Yu
- Jian Li
View Affiliations

Affiliations: Center for Gene Diagnosis, Zhongnan Hospital, Wuhan University, Wuhan, Hubei 430071, P.R. China, Life Science Research Institute of Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Life Science Research Institute of Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Medicine, Shenzhen Family Planning Service Center, Shenzhen, Guangdong 518028, P.R. China, The Key Laboratory of Geriatrics, Beijing Institute of Geriatrics and Beijing Hospital, Ministry of Health, Dongdan, Beijing 100730, P.R. China
Published online on: September 18, 2013 https://doi.org/10.3892/mmr.2013.1691
Pages: 1472-1478

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

MicroRNAs (miRNAs) are small non‑coding RNAs that are important in the pathogenesis of multiple diseases and, therefore, may represent a novel class of targets for therapeutic intervention. However, like the majority of oligonucleotide‑based strategies, there are obstacles to their clinical application, including poor cellular uptake due to the low permeability of the cell membrane to negatively charged molecules. MPG is a 27‑residue peptide vector which contains a hydrophobic domain derived from the fusion sequence of HIV-1 gp41 and a hydrophilic domain derived from the nuclear localization sequence of SV40 T-antigen. MPG is one of the most promising tools for the non‑invasive cellular import of oligonucleotides and analogs. In the present study, a non‑covalent peptide‑based strategy was used for the efficient delivery of the miRNA‑122 (miR‑122) mimic and inhibitor into mouse liver cell lines, mouse primary hepatocytes and C. elegans, without any associated cytotoxicity. Moreover, high‑performance liquid chromatography analysis determined that MPG and MPGΔNLS delivered the miR‑122 mimic and inhibitor into mouse liver cells and effectively regulated cholesterol levels. The results demonstrated that MPG family members may be used for the efficient delivery of miR‑122 to regulate cholesterol metabolism, and that this cell‑penetrating peptide‑based technology may be beneficial for further biological applications of RNA therapeutics in vivo.

View Figures

View References

1	Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
2	Lee Y, Ahn C, Han J, et al: The nuclear RNase III Drosha initiates microRNA processing. Nature. 425:415–419. 2003. View Article : Google Scholar : PubMed/NCBI
3	Yi R, Qin Y, Macara IG and Cullen BR: Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs. Genes Dev. 17:3011–3016. 2003. View Article : Google Scholar : PubMed/NCBI
4	Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI
5	Yekta S, Shih IH and Bartel DP: MicroRNA-directed cleavage of HOXB8 mRNA. Science. 304:594–596. 2004. View Article : Google Scholar : PubMed/NCBI
6	Hildebrandt-Eriksen ES, Aarup V, Persson R, Hansen HF, Munk ME and Orum H: A locked nucleic acid oligonucleotide targeting microRNA 122 is well-tolerated in cynomolgus monkeys. Nucleic Acid Ther. 22:152–161. 2012.PubMed/NCBI
7	Elmén J, Lindow M, Schutz S, et al: LNA-mediated microRNA silencing in non-human primates. Nature. 452:896–899. 2008.PubMed/NCBI
8	Heitz F, Morris MC and Divita G: Twenty years of cell-penetrating peptides: from molecular mechanisms to therapeutics. Br J Pharmacol. 157:195–206. 2009.PubMed/NCBI
9	Zatsepin TS, Turner JJ, Oretskaya TS and Gait MJ: Conjugates of oligonucleotides and analogues with cell penetrating peptides as gene silencing agents. Curr Pharm Des. 11:3639–3654. 2005. View Article : Google Scholar : PubMed/NCBI
10	El-Andaloussi S, Holm T and Langel U: Cell-penetrating peptides: mechanisms and applications. Curr Pharm Des. 11:3597–3611. 2005. View Article : Google Scholar : PubMed/NCBI
11	Joliot A and Prochiantz A: Transduction peptides: from technology to physiology. Nat Cell Biol. 6:189–196. 2004. View Article : Google Scholar : PubMed/NCBI
12	Torchilin VP: Tat peptide-mediated intracellular delivery of pharmaceutical nanocarriers. Adv Drug Deliv Rev. 60:548–558. 2008. View Article : Google Scholar : PubMed/NCBI
13	Deshayes S, Morris M, Heitz F and Divita G: Delivery of proteins and nucleic acids using a non-covalent peptide-based strategy. Adv Drug Deliv Rev. 60:537–547. 2008. View Article : Google Scholar : PubMed/NCBI
14	Deshayes S, Morris MC, Divita G and Heitz F: Cell-penetrating peptides: tools for intracellular delivery of therapeutics. Cell Mol Life Sci. 62:1839–1849. 2005. View Article : Google Scholar : PubMed/NCBI
15	Snyder EL and Dowdy SF: Recent advances in the use of protein transduction domains for the delivery of peptides, proteins and nucleic acids in vivo. Expert Opin Drug Deliv. 2:43–51. 2005. View Article : Google Scholar : PubMed/NCBI
16	Eguchi A and Dowdy SF: siRNA delivery using peptide transduction domains. Trends Pharmacol Sci. 30:341–345. 2009. View Article : Google Scholar : PubMed/NCBI
17	Simeoni F, Morris MC, Heitz F and Divita G: Insight into the mechanism of the peptide-based gene delivery system MPG: implications for delivery of siRNA into mammalian cells. Nucleic Acids Res. 31:2717–2724. 2003. View Article : Google Scholar : PubMed/NCBI
18	Yoo JW, Hong SW, Kim S and Lee DK: Inflammatory cytokine induction by siRNAs is cell type- and transfection reagent-specific. Biochem Biophys Res Commun. 347:1053–1058. 2006. View Article : Google Scholar : PubMed/NCBI
19	Crombez L, Charnet A, Morris MC, Aldrian-Herrada G, Heitz F and Divita G: A non-covalent peptide-based strategy for siRNA delivery. Biochem Soc Trans. 35:44–46. 2007. View Article : Google Scholar : PubMed/NCBI
20	Lundberg P, El-Andaloussi S, Sutlu T, Johansson H and Langel U: Delivery of short interfering RNA using endosomolytic cell-penetrating peptides. FASEB J. 21:2664–2671. 2007. View Article : Google Scholar : PubMed/NCBI
21	Simeoni F, Morris MC, Heitz F and Divita G: Peptide-based strategy for siRNA delivery into mammalian cells. Methods Mol Biol. 309:251–260. 2005.PubMed/NCBI
22	Morris MC, Chaloin L, Méry J, Heitz F and Divita G: A novel potent strategy for gene delivery using a single peptide vector as a carrier. Nucleic Acids Res. 27:3510–3517. 1999. View Article : Google Scholar : PubMed/NCBI
23	Morris KV, Chan SW, Jacobsen SE and Looney DJ: Small interfering RNA-induced transcriptional gene silencing in human cells. Science. 305:1289–1292. 2004. View Article : Google Scholar : PubMed/NCBI
24	Nguyen QN, Chavli RV, Marques JT, et al: Light controllable siRNAs regulate gene suppression and phenotypes in cells. Biochim Biophys Acta. 1758:394–403. 2006. View Article : Google Scholar : PubMed/NCBI
25	Esau C, Davis S, Murray SF, et al: miR-122 regulation of lipid metabolism revealed by in vivo antisense targeting. Cell Metab. 3:87–98. 2006. View Article : Google Scholar : PubMed/NCBI
26	Liu GT, Carrazana EJ, Macklis JD and Mikati MA: Delayed oculogyric crises associated with striatocapsular infarction. J Clin Neuroophthalmol. 11:198–201. 1991.PubMed/NCBI
27	Esau CC: Inhibition of microRNA with antisense oligonucleotides. Methods. 44:55–60. 2008. View Article : Google Scholar : PubMed/NCBI
28	Lanford RE, Hildebrandt-Eriksen ES, Petri A, et al: Therapeutic silencing of microRNA-122 in primates with chronic hepatitis C virus infection. Science. 327:198–201. 2010. View Article : Google Scholar : PubMed/NCBI
29	Méry J, Granier C, Juin M and Brugidou J: Disulfide linkage to polyacrylic resin for automated Fmoc peptide synthesis. Immunochemical applications of peptide resins and mercaptoamide peptides. Int J Pept Protein Res. 42:44–52. 1993.PubMed/NCBI
30	Salonpää P, Pelkonen O, Kojo A, Pasanen M, Negishi M and Raunio H: Cytochrome P4502A5 expression and inducibility by phenobarbital is modulated by cAMP in mouse primary hepatocytes. Biochem Biophys Res Commun. 205:631–637. 1994.PubMed/NCBI
31	Dong J, Guo H, Yang R, et al: Serum LDL- and HDL-cholesterol determined by ultracentrifugation and HPLC. J Lipid Res. 52:383–388. 2011. View Article : Google Scholar : PubMed/NCBI
32	Krützfeldt J, Rajewsky N, Braich R, et al: Silencing of microRNAs in vivo with ‘antagomirs’. Nature. 438:685–689. 2005.
33	Crombez L, Morris MC, Heitz F and Divita G: A non-covalent peptide-based strategy for ex vivo and in vivo oligonucleotide delivery. Methods Mol Biol. 764:59–73. 2011. View Article : Google Scholar : PubMed/NCBI
34	Crombez L, Morris MC, Dufort S, et al: Targeting cyclin B1 through peptide-based delivery of siRNA prevents tumour growth. Nucleic Acids Res. 37:4559–4569. 2009. View Article : Google Scholar : PubMed/NCBI
35	Jones S and Howl J: Applications of cell-penetrating peptides as signal transduction modulators for the selective induction of apoptosis. Methods Mol Biol. 683:291–303. 2011. View Article : Google Scholar : PubMed/NCBI
36	Kimber MJ, McKinney S, McMaster S, Day TA, Fleming CC and Maule AG: flp gene disruption in a parasitic nematode reveals motor dysfunction and unusual neuronal sensitivity to RNA interference. FASEB J. 21:1233–1243. 2007. View Article : Google Scholar : PubMed/NCBI