Non-covalently functionalized single-walled carbon nanotube for topical siRNA delivery into melanoma.

RNAi can specifically regulate gene expression, but efficient delivery of siRNA in vivo is difficult while it has been shown that modified carbon nanotubes (CNT) protect siRNA, facilitate entry into cells and enhance transdermal drugs delivery. Single-walled carbon nanotubes (SWCNT) were functionalized non-covalently with succinated polyethyleimine (PEI-SA). In this study, the water soluble CNT, PEI-SA/CNT (IS/C) were isolated and characterized, the gene silencing induced by IS/C/siRNA complexes was achieved in vitro in B16-F10 cells. In vivo delivery was topically applied to shaved mouse skin, as well as topically to a C57BL/6 mice melanoma model. We found significant uptake of Cy3-labeled siRNA specific to Braf (siBraf) and gene silencing in the tumor tissue. Treatment with IS/C/siBraf resulted in attenuation of tumor growth over a 25-day period. This new delivery method has provided a new possibility for future siRNA delivery and therapy, which providing insight for the potential application and development of CNT-based siRNA delivery.

[1]  D. Hudson,et al.  Delay in the diagnosis of cutaneous malignant melanoma. A prospective study in 250 patients , 1991, Cancer.

[2]  A. Eychène,et al.  Identification of signalling proteins interacting with B-Raf in the yeast two-hybrid system. , 1996, Oncogene.

[3]  M. Prato,et al.  Antitumor activity and prolonged survival by carbon-nanotube-mediated therapeutic siRNA silencing in a human lung xenograft model. , 2009, Small.

[4]  K. Kanamura,et al.  Noninvasive delivery of siRNA into the epidermis by iontophoresis using an atopic dermatitis-like model rat. , 2010, International journal of pharmaceutics.

[5]  H. Ananthaswamy,et al.  Genomic alterations in spontaneous and carcinogen-induced murine melanoma cell lines , 2004, Oncogene.

[6]  Giorgia Pastorin,et al.  Crucial Functionalizations of Carbon Nanotubes for Improved Drug Delivery: A Valuable Option? , 2009, Pharmaceutical Research.

[7]  M. Azuma,et al.  Topical application of cream-emulsified CD86 siRNA ameliorates allergic skin disease by targeting cutaneous dendritic cells. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[8]  V. Popov.  Carbon Nanotubes: Properties and Applications , 2006 .

[9]  A. Cuschieri,et al.  Physicochemical properties affecting cellular uptake of carbon nanotubes. , 2010, Nanomedicine.

[10]  Mingwu Shen,et al.  Effect of surface charge of polyethyleneimine-modified multiwalled carbon nanotubes on the improvement of polymerase chain reaction. , 2011, Nanoscale.

[11]  R. Nemanich,et al.  Multi-walled carbon nanotube interactions with human epidermal keratinocytes. , 2005, Toxicology letters.

[12]  M. Hung,et al.  Nonviral vectors for gene therapy , 1999 .

[13]  Robert Langer,et al.  Knocking down barriers: advances in siRNA delivery , 2009, Nature Reviews Drug Discovery.

[14]  N. K. Jain,et al.  A review of nanocarriers for the delivery of small interfering RNA. , 2012, Biomaterials.

[15]  A. Pandit,et al.  RNA interference therapy via functionalized scaffolds. , 2011, Advanced drug delivery reviews.

[16]  M. O'connell,et al.  Carbon Nanotubes Properties and Applications , 2006 .

[17]  H. Krug,et al.  Oops they did it again! Carbon nanotubes hoax scientists in viability assays. , 2006, Nano letters.

[18]  Yuan Zhang,et al.  Delivery of Telomerase Reverse Transcriptase Small Interfering RNA in Complex with Positively Charged Single-Walled Carbon Nanotubes Suppresses Tumor Growth , 2006, Clinical Cancer Research.

[19]  Zhuang Liu,et al.  Functionalization of carbon nanotubes via cleavable disulfide bonds for efficient intracellular delivery of siRNA and potent gene silencing. , 2005, Journal of the American Chemical Society.

[20]  Fotios Papadimitrakopoulos,et al.  Carbon nanotubes for transdermal drug delivery , 2010, Journal of microencapsulation.

[21]  M. Shim,et al.  Noncovalent functionalization of carbon nanotubes for highly specific electronic biosensors , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  D. Barford,et al.  Wild-type and mutant B-RAF activate C-RAF through distinct mechanisms involving heterodimerization. , 2005, Molecular cell.

[23]  O. Danos,et al.  Polyethylenimine‐mediated gene delivery: a mechanistic study , 2001, The journal of gene medicine.

[24]  D. Putnam,et al.  Polymer systems for gene delivery - Past, present, and future , 2007 .

[25]  Maurizio Prato,et al.  Enhanced cellular internalization and gene silencing with a series of cationic dendron‐multiwalled carbon nanotube:siRNA complexes , 2010, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[26]  D. Elder.  Tumor progression, early diagnosis and prognosis of melanoma. , 1999, Acta oncologica.

[27]  G. Robertson,et al.  Targeting V600EB-Raf and Akt3 using nanoliposomal-small interfering RNA inhibits cutaneous melanocytic lesion development. , 2008, Cancer research.

[28]  Maurizio Prato,et al.  Synthesis and characterization of a carbon nanotube-dendron series for efficient siRNA delivery. , 2009, Journal of the American Chemical Society.

[29]  A. Jorio,et al.  Highly efficient siRNA delivery system into human and murine cells using single-wall carbon nanotubes , 2010, Nanotechnology.

[30]  C. Garbe,et al.  Diagnosis and treatment of cutaneous melanoma: state of the art 2006. , 2007, Melanoma research.

[31]  田波 秀朗.  Involvement of overexpressed wild-type BRAF in the growth of malignant melanoma cell lines , 2004 .

[32]  Jianzhu Chen,et al.  Cross-linked Small Polyethylenimines: While Still Nontoxic, Deliver DNA Efficiently to Mammalian Cells in Vitro and in Vivo , 2005, Pharmaceutical Research.

[33]  Hongjie Dai,et al.  siRNA delivery into human T cells and primary cells with carbon-nanotube transporters. , 2007, Angewandte Chemie.

[34]  R. Smalley,et al.  Reversible water-solubilization of single-walled carbon nanotubes by polymer wrapping , 2001 .

[35]  Stanislaus S. Wong,et al.  Functionalized single-walled carbon nanotubes as rationally designed vehicles for tumor-targeted drug delivery. , 2008, Journal of the American Chemical Society.

[36]  Zhixin Guo,et al.  Carbon Nanotube Delivery of the GFP Gene into Mammalian Cells , 2006, Chembiochem : a European journal of chemical biology.

[37]  Young-Seak Lee,et al.  The effect of carbon nanotubes on drug delivery in an electro-sensitive transdermal drug delivery system. , 2010, Biomaterials.

[38]  A. Brunet,et al.  Constitutively active mutants of MAP kinase kinase (MEK1) induce growth factor-relaxation and oncogenicity when expressed in fibroblasts. , 1994, Oncogene.

[39]  B. Hinds,et al.  Programmable transdermal drug delivery of nicotine using carbon nanotube membranes , 2010, Proceedings of the National Academy of Sciences.

[40]  M. Ogris,et al.  Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. , 2001, Nucleic acids research.

[41]  K. Flaherty,et al.  Inhibition of mutated, activated BRAF in metastatic melanoma. , 2010, The New England journal of medicine.

[42]  Wim E Hennink,et al.  SiRNA delivery with functionalized carbon nanotubes. , 2011, International journal of pharmaceutics.

[43]  Ning Wang,et al.  Biofunctionalized nanoneedles for the direct and site-selective delivery of probes into living cells. , 2011, Biochimica et biophysica acta.

[44]  STEFAN C. Weiss.  Conventional topical delivery systems , 2011, Dermatologic therapy.

[45]  M. Prato,et al.  Chemistry of carbon nanotubes. , 2006, Chemical reviews.

[46]  Maurizio Prato,et al.  Double functionalization of carbon nanotubes for multimodal drug delivery. , 2006, Chemical communications.

[47]  Zhuang Liu,et al.  Drug delivery with carbon nanotubes for in vivo cancer treatment. , 2008, Cancer research.

[48]  S. MacNeil,et al.  Transdermal drug delivery: from micro to nano. , 2012, Nanoscale.

[49]  G. Zuber,et al.  Polyethylenimine-carbon nanotube nanohybrids for siRNA-mediated gene silencing at cellular level. , 2011, Nanoscale.

[50]  K. Leong,et al.  Polyethylenimine-grafted multiwalled carbon nanotubes for secure noncovalent immobilization and efficient delivery of DNA. , 2005, Angewandte Chemie.

[51]  K. Jain.  Advances in use of functionalized carbon nanotubes for drug design and discovery , 2012, Expert opinion on drug discovery.

[52]  A. Spadaro,et al.  Liquid crystalline phase nanodispersions enable skin delivery of siRNA. , 2013, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[53]  E. Wagner,et al.  Simple modifications of branched PEI lead to highly efficient siRNA carriers with low toxicity. , 2008, Bioconjugate chemistry.