title: Plasma hydrogenated cationic detonation nanodiamonds efficiently deliver to human cells in culture functional siRNA targeting the Ewing sarcoma junction oncogene.
abstract: The expression of a defective gene can lead to major cell dysfunctions among which cell proliferation and tumor formation. One promising therapeutic strategy consists in silencing the defective gene using small interfering RNA (siRNA). In previous publications we showed that diamond nanocrystals (ND) of primary size 35 nm, rendered cationic by polyethyleneimine-coating, can efficiently deliver siRNA into cell, which further block the expression of EWS/FLI-1 oncogene in a Ewing sarcoma disease model. However, a therapeutic application of such nanodiamonds requires their elimination by the organism, particularly in urine, which is impossible for 35 nm particles. Here, we report that hydrogenated cationic nanodiamonds of primary size 7 nm (ND-H) have also a high affinity for siRNA and are capable of delivering them in cells. With siRNA/ND-H complexes, we measured a high inhibition efficacy of EWS/FLI-1 gene expression in Ewing sarcoma cell line. Electron microscopy investigations showed ND-H in endocytosis compartments, and especially in macropinosomes from which they can escape before siRNA degradation occurred. In addition, the association of EWS/FLI-1 silencing by the siRNA/ND-H complex with a vincristine treatment yielded a potentiation of the toxic effect of this chemotherapeutic drug. Therefore ND-H appears as a promising delivery agent in anti-tumoral gene therapy.
authors: Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR 8203 CNRS, Université Paris Sud, Institut Gustave Roussy, 94805 Villejuif, France. Electronic address: jean-remi.bertrand@gustaveroussy.fr.
authors: Jean-Rémi
authors: JR
authors: Bertrand
authors: Signalisation, Noyaux et Innovations en Cancérologie, UMR 8126 CNRS, Université Paris Sud, Institut Gustave Roussy, 94805 Villejuif, France.
authors: Catherine
authors: C
authors: Pioche-Durieu
authors: Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR 8203 CNRS, Université Paris Sud, Institut Gustave Roussy, 94805 Villejuif, France.
authors: Juan
authors: J
authors: Ayala
authors: CEA, LIST, Diamond Sensors Laboratory, F-91191 Gif-sur-Yvette, France.
authors: Tristan
authors: T
authors: Petit
authors: CEA, LIST, Diamond Sensors Laboratory, F-91191 Gif-sur-Yvette, France.
authors: Hugues A
authors: HA
authors: Girard
authors: Laboratoire de Vectorologie et Thérapeutiques Anticancéreuses, UMR 8203 CNRS, Université Paris Sud, Institut Gustave Roussy, 94805 Villejuif, France.
authors: Claude P
authors: CP
authors: Malvy
authors: Signalisation, Noyaux et Innovations en Cancérologie, UMR 8126 CNRS, Université Paris Sud, Institut Gustave Roussy, 94805 Villejuif, France.
authors: Eric
authors: E
authors: Le Cam
authors: Laboratoire Aimé Cotton, CNRS, Université Paris Sud and ENS Cachan, 91405 Orsay, France. Electronic address: francois.treussart@ens-cachan.fr.
authors: François
authors: F
authors: Treussart
authors: CEA, LIST, Diamond Sensors Laboratory, F-91191 Gif-sur-Yvette, France. Electronic address: jean-charles.arnault@cea.fr.
authors: Jean-Charles
authors: JC
authors: Arnault
mesh_terms: D002412:Cations; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D004705:Endocytosis / Q000187:drug effects; D005453:Fluorescence; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018014:Gene Transfer Techniques; D006801:Humans; D006865:Hydrogenation; D058612:Nanodiamonds / Q000737:chemistry* / Q000648:ultrastructure; D015514:Oncogene Proteins, Fusion / Q000235:genetics* / Q000378:metabolism; D058626:Plasma Gases / Q000737:chemistry*; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D034802:RNA-Binding Protein EWS / Q000235:genetics* / Q000378:metabolism; D012512:Sarcoma, Ewing / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D014750:Vincristine / Q000494:pharmacology
chemical_list: D002412:Cations; C106031:EWS-FLI fusion protein; D058612:Nanodiamonds; D015514:Oncogene Proteins, Fusion; D058626:Plasma Gases; D051789:Proto-Oncogene Protein c-fli-1; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D034802:RNA-Binding Protein EWS; D014750:Vincristine
keywords: Drug delivery; Ewing sarcoma; Gene therapy; Nanodiamonds; Nanomedicine; Plasma hydrogenation; siRNA