TY - JOUR
T1 - Gene silencing via RNAi and siRNA quantification in tumor tissue using MEND, a liposomal siRNA delivery system
AU - Sakurai, Yu
AU - Hatakeyama, Hiroto
AU - Sato, Yusuke
AU - Hyodo, Mamoru
AU - Akita, Hidetaka
AU - Harashima, Hideyoshi
N1 - Funding Information:
This study was supported in part by a grant-in-aid for Young Scientists (B) from the Japan Society for the Promotion of Science (JSPS), the Special Education and Research Expenses of the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan, a grant-in-aid for Scientific Research on Innovative Areas “Nanomedicine Molecular Science” (No. 2306) from MEXT of Japan, and by a grant for Industrial Technology Research from New Energy and Industrial Technology Development Organization (NEDO). The authors declare no conflict of interest.
PY - 2013/6
Y1 - 2013/6
N2 - Small interfering RNA (siRNA) would be predicted to function as a cancer drug, but an efficient siRNA delivery system is required for clinical development. To address this issue, we developed a liposomal siRNA carrier, a multifunctional envelope-type nanodevice (MEND). We previously reported that a MEND composed of a pH-sensitive cationic lipid, YSK05, showed significant knockdown in both in vitro and in tumor tissue by intratumoral injection. Here, we report on the development of an in vivo siRNA delivery system that is delivered by systemic injection and an analysis of the pharmacokinetics of an intravenously administered siRNA molecule in tumor tissue. Tumor delivery of siRNA was quantified by means of stem-loop primer quantitative reverse transcriptase PCR (qRT-PCR) method. PEGylation of the YSK-MEND results in the increase in the accumulation of siRNA in tumor tissue from 0.0079% ID/g tumor to 1.9% ID/g tumor. The Administration of the MEND (3 mg siRNA/kg body weight) showed about a 50% reduction in the target gene mRNA and protein. Moreover, we verified the induction of RNA interference by 5′ RACE-PCR method. The collective results reported here indicate that an siRNA carrier was developed that can deliver siRNA to a target cell in tumor tissue through an improved siRNA bioavailability.
AB - Small interfering RNA (siRNA) would be predicted to function as a cancer drug, but an efficient siRNA delivery system is required for clinical development. To address this issue, we developed a liposomal siRNA carrier, a multifunctional envelope-type nanodevice (MEND). We previously reported that a MEND composed of a pH-sensitive cationic lipid, YSK05, showed significant knockdown in both in vitro and in tumor tissue by intratumoral injection. Here, we report on the development of an in vivo siRNA delivery system that is delivered by systemic injection and an analysis of the pharmacokinetics of an intravenously administered siRNA molecule in tumor tissue. Tumor delivery of siRNA was quantified by means of stem-loop primer quantitative reverse transcriptase PCR (qRT-PCR) method. PEGylation of the YSK-MEND results in the increase in the accumulation of siRNA in tumor tissue from 0.0079% ID/g tumor to 1.9% ID/g tumor. The Administration of the MEND (3 mg siRNA/kg body weight) showed about a 50% reduction in the target gene mRNA and protein. Moreover, we verified the induction of RNA interference by 5′ RACE-PCR method. The collective results reported here indicate that an siRNA carrier was developed that can deliver siRNA to a target cell in tumor tissue through an improved siRNA bioavailability.
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U2 - 10.1038/mt.2013.57
DO - 10.1038/mt.2013.57
M3 - Article
C2 - 23568259
AN - SCOPUS:84878567681
VL - 21
SP - 1195
EP - 1203
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 6
ER -
