PMID- 28528119
OWN - NLM
STAT- MEDLINE
DCOM- 20180124
LR  - 20180221
IS  - 1878-7568 (Electronic)
IS  - 1742-7061 (Linking)
VI  - 58
DP  - 2017 Aug
TI  - Autophagy plays a dual role during intracellular siRNA delivery by lipoplex and 
      polyplex nanoparticles.
PG  - 196-204
LID - S1742-7061(17)30329-X [pii]
LID - 10.1016/j.actbio.2017.05.038 [doi]
AB  - Growing evidence indicates that autophagy plays a vital role during intracellular 
      DNA delivery mediated by lipoplex and polyplex nanoparticles. However, autophagy 
      in intracellular siRNA delivery has not been well understood. In this study, 
      lipofectamine 2000 and chitosan were used to formulate lipoplex and polyplex with 
      siRNA for systematically investigating the interplay between siRNA delivery and 
      autophagy. After transfection of H1299 cells with lipoplex and polyplex, the 
      number of autophagic vacuoles was increased significantly indicated by the 
      accumulation of monodansylcadaverine (MDC) staining. Western blot revealed that 
      the LC3-II expression was significantly increased after transfection, whereas 
      p-mTOR expression was not influenced apparently. In addition, small-molecule 
      autophagy modulators significantly affected transfection efficiency. 
      Specifically, the mTOR-dependent autophagy inducer rapamycin enhanced the 
      knockdown efficiency of both lipoplex and polyplex, whereas mTOR-dependent 
      autophagy inhibitor 3-methyladenine (3-MA) suppressed their silencing efficiency. 
      On the contrary, mTOR-independent autophagy inducer LiBr decreased whereas 
      mTOR-independent autophagy inhibitor thapsigargin (TG) increased the knockdown 
      efficacy. Immunofluorescence staining showed that siRNA was partially 
      co-localized with autophagosomes and the percentage of co-localized siRNA was 
      significantly affected by autophagy modulators in the opposite trend of gene 
      knockdown efficacy. In conclusion, our study suggests that autophagy plays an 
      important role during the intracellular siRNA trafficking mediated by both 
      lipoplex and polyplex. Modulating autophagy process will result in distinct 
      knockdown efficiency, which may be applied as a potential convenient way for 
      improving siRNA delivery efficacy. STATEMENT OF SIGNIFICANCE: Although tremendous 
      effects has been made in the development of non-viral siRNA delivery systems, the 
      intracellular siRNA trafficking has not been elucidated clearly. In this study, 
      we systematically investigated the relationship between autophagy and 
      intracellular siRNA delivery. We found that the non-viral siRNA delivery by both 
      lipoplex and polyplex could induce mTOR-independent autophagy response. More 
      interestingly, knockdown efficiency of both lipoplex and polyplex could be 
      modulated with different autophagy regulators. Specifically, the mTOR-dependent 
      autophagy inducer rapamycin enhances the knockdown efficiency of both lipoplex 
      and polyplex, whereas mTOR-dependent autophagy inhibitor 3-methyladenine 
      suppresses their silencing efficiency. On the contrary, mTOR-independent 
      autophagy inducer lithium bromide decreases, whereas mTOR-independent autophagy 
      inhibitor thapsigargin increases the knockdown efficacy. These findings suggest 
      that the mTOR-dependent and -independent autophagy play a distinct role in the 
      intracellular siRNA trafficking. Furthermore, co-administration with proper 
      autophagy regulators could be potential convenient method to modulate siRNA 
      transfection efficacy.
CI  - Copyright (c) 2017. Published by Elsevier Ltd.
FAU - Song, Wen
AU  - Song W
AD  - State Key Laboratory of Military Stomatology & National Clinical Research Center 
      for Oral Diseases & Shaanxi Key Laboratory of Oral Diseases, Department of 
      Prosthodontics, School of Stomatology, The Fourth Military Medical University, 
      Xi'an 710032, China; Interdisciplinary Nanoscience Center (iNANO), Aarhus 
      University, Aarhus 8000, Denmark.
FAU - Ma, Zhiwei
AU  - Ma Z
AD  - State Key Laboratory of Military Stomatology & National Clinical Research Center 
      for Oral Diseases & Shaanxi Key Laboratory of Oral Diseases, Department of 
      Periodontology, School of Stomatology, The Fourth Military Medical University, 
      Xi'an 710032, China.
FAU - Zhang, Yumei
AU  - Zhang Y
AD  - State Key Laboratory of Military Stomatology & National Clinical Research Center 
      for Oral Diseases & Shaanxi Key Laboratory of Oral Diseases, Department of 
      Prosthodontics, School of Stomatology, The Fourth Military Medical University, 
      Xi'an 710032, China. Electronic address: wqtzym@fmmu.edu.cn.
FAU - Yang, Chuanxu
AU  - Yang C
AD  - Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus 8000, 
      Denmark. Electronic address: chuanxuyang@inano.au.dk.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170517
PL  - England
TA  - Acta Biomater
JT  - Acta biomaterialia
JID - 101233144
RN  - 0 (RNA, Small Interfering)
SB  - IM
MH  - Autophagy/*drug effects/genetics
MH  - Cell Line, Tumor
MH  - Drug Delivery Systems/*methods
MH  - Humans
MH  - *Nanoparticles/chemistry/therapeutic use
MH  - *RNA, Small Interfering/chemistry/pharmacokinetics/pharmacology
OTO - NOTNLM
OT  - Autophagy
OT  - Lipoplex
OT  - Nanoparticles
OT  - Polyplex
OT  - siRNA delivery
EDAT- 2017/05/22 06:00
MHDA- 2018/01/25 06:00
CRDT- 2017/05/22 06:00
PHST- 2016/12/29 00:00 [received]
PHST- 2017/05/07 00:00 [revised]
PHST- 2017/05/15 00:00 [accepted]
PHST- 2017/05/22 06:00 [pubmed]
PHST- 2018/01/25 06:00 [medline]
PHST- 2017/05/22 06:00 [entrez]
AID - S1742-7061(17)30329-X [pii]
AID - 10.1016/j.actbio.2017.05.038 [doi]
PST - ppublish
SO  - Acta Biomater. 2017 Aug;58:196-204. doi: 10.1016/j.actbio.2017.05.038. Epub 2017 
      May 17.