Targeted exosome-mediated delivery of opioid receptor Mu siRNA for the treatment of morphine relapse.

Yuchen Liu,Ke Zen,Yuda Chen,Zhengya Liu,Junfeng Zhang,Danping Chu,Ling Jin,Waner Jiang,Jing Li,Chenfei Tian,Yujing Zhang,Xiaohong Jiang,Geyu Zhou,Zhanzhao Yang,Xi Chen,Yu Zhou,Dameng Li,De‐Xing Hou ,Xi-Han Li ,Chenyu Zhang

doi:10.1038/srep17543

Abstract

Cell-derived exosomes have been demonstrated to be efficient carriers of small RNAs to neighbouring or distant cells, highlighting the preponderance of exosomes as carriers for gene therapy over other artificial delivery tools. In the present study, we employed modified exosomes expressing the neuron-specific rabies viral glycoprotein (RVG) peptide on the membrane surface to deliver opioid receptor mu (MOR) siRNA into the brain to treat morphine addiction. We found that MOR siRNA could be efficiently packaged into RVG exosomes and was associated with argonaute 2 (AGO2) in exosomes. These exosomes efficiently and specifically delivered MOR siRNA into Neuro2A cells and the mouse brain. Functionally, siRNA-loaded RVG exosomes significantly reduced MOR mRNA and protein levels. Surprisingly, MOR siRNA delivered by the RVG exosomes strongly inhibited morphine relapse via the down-regulation of MOR expression levels. In conclusion, our results demonstrate that targeted RVG exosomes can efficiently transfer siRNA to the central nervous system and mediate the treatment of morphine relapse by down-regulating MOR expression levels. Our study provides a brand new strategy to treat drug relapse and diseases of the central nervous system.

Highlights

Cell-derived exosomes have been demonstrated to be efficient carriers of small RNAs to neighbouring or distant cells, highlighting the preponderance of exosomes as carriers for gene therapy over other artificial delivery tools
The transmission electron microscopy (TEM) photographs showed that the exosomes presented normal morphological characteristics, with a diameter of approximately 90 nm, and that each vesicle was surrounded by a double-layer membrane; the NTA results showed that the diameter of majority of particles are 85 nm
The following study further demonstrated that exogenous small-interfering RNA (siRNA) transfected into cells could be packaged by exosomes and delivered into recipient cells to regulates gene silencing, indicating that exosomes can serve as siRNA delivery vesicles in gene therapy for cancer and other diseases[12,13,14]

Summary

Introduction

Cell-derived exosomes have been demonstrated to be efficient carriers of small RNAs to neighbouring or distant cells, highlighting the preponderance of exosomes as carriers for gene therapy over other artificial delivery tools. We employed modified exosomes expressing the neuron-specific rabies viral glycoprotein (RVG) peptide on the membrane surface to deliver opioid receptor mu (MOR) siRNA into the brain to treat morphine addiction. MOR siRNA delivered by the RVG exosomes strongly inhibited morphine relapse via the down-regulation of MOR expression levels. Our results demonstrate that targeted RVG exosomes can efficiently transfer siRNA to the central nervous system and mediate the treatment of morphine relapse by down-regulating MOR expression levels. We utilised RVG exosomes loaded with opioid receptor Mu (MOR) siRNA to treat drug addiction via down-regulating the expression of MOR, which is the primary target for opioid analgesics used clinically, including morphine, fentanyl and methadone, and is involved in the primary reinforcing effects of and the addiction to opiates. Our study provides a brand new strategy for treating drug addiction

Methods

Results

Conclusion