Delivery of SiRNA to the Mouse Brain by Systemic Injection of Targeted Exosomes

Overview

Journal Nat Biotechnol

Specialty Biotechnology

Date 2011 Mar 23

PMID 21423189

Citations 2111

Authors

Lydia Alvarez-Erviti

Yiqi Seow

HaiFang Yin

Corinne Betts

Samira Lakhal

Matthew J A Wood

Affiliations

Soon will be listed here.

Abstract

To realize the therapeutic potential of RNA drugs, efficient, tissue-specific and nonimmunogenic delivery technologies must be developed. Here we show that exosomes-endogenous nano-vesicles that transport RNAs and proteins-can deliver short interfering (si)RNA to the brain in mice. To reduce immunogenicity, we used self-derived dendritic cells for exosome production. Targeting was achieved by engineering the dendritic cells to express Lamp2b, an exosomal membrane protein, fused to the neuron-specific RVG peptide. Purified exosomes were loaded with exogenous siRNA by electroporation. Intravenously injected RVG-targeted exosomes delivered GAPDH siRNA specifically to neurons, microglia, oligodendrocytes in the brain, resulting in a specific gene knockdown. Pre-exposure to RVG exosomes did not attenuate knockdown, and non-specific uptake in other tissues was not observed. The therapeutic potential of exosome-mediated siRNA delivery was demonstrated by the strong mRNA (60%) and protein (62%) knockdown of BACE1, a therapeutic target in Alzheimer's disease, in wild-type mice.

Citing Articles

Prospect of extracellular vesicles in tumor immunotherapy.

Xia W, Tan Y, Liu Y, Xie N, Zhu H Front Immunol. 2025; 16:1525052.

PMID: 40078996 PMC: 11897508. DOI: 10.3389/fimmu.2025.1525052.

Landscape of small nucleic acid therapeutics: moving from the bench to the clinic as next-generation medicines.

Liu M, Wang Y, Zhang Y, Hu D, Tang L, Zhou B Signal Transduct Target Ther. 2025; 10(1):73.

PMID: 40059188 PMC: 11891339. DOI: 10.1038/s41392-024-02112-8.

Injured Cardiac Tissue-Targeted Delivery of TGFβ1 siRNA by FAP Aptamer-Functionalized Extracellular Vesicles Promotes Cardiac Repair.

Kang J, Mun D, Park M, Yoo G, Kim H, Yun N Int J Nanomedicine. 2025; 20:2575-2592.

PMID: 40046817 PMC: 11881639. DOI: 10.2147/IJN.S497428.

An anti-CD19-exosome delivery system navigates the blood-brain barrier for targeting of central nervous system lymphoma.

Zhao M, Li Q, Chai Y, Rong R, He L, Zhang Y J Nanobiotechnology. 2025; 23(1):173.

PMID: 40045315 PMC: 11881385. DOI: 10.1186/s12951-025-03238-9.

Current challenges surrounding exosome treatments.

Tzng E, Bayardo N, Yang P Extracell Vesicle. 2025; 2:100023.

PMID: 40027080 PMC: 11870656. DOI: 10.1016/j.vesic.2023.100023.

References

Aguzzi A, OConnor T . Protein aggregation diseases: pathogenicity and therapeutic perspectives. Nat Rev Drug Discov. 2010; 9(3):237-48. DOI: 10.1038/nrd3050. View

Singer O, Marr R, Rockenstein E, Crews L, Coufal N, Gage F . Targeting BACE1 with siRNAs ameliorates Alzheimer disease neuropathology in a transgenic model. Nat Neurosci. 2005; 8(10):1343-9. DOI: 10.1038/nn1531. View

Laird F, Cai H, Savonenko A, Farah M, He K, Melnikova T . BACE1, a major determinant of selective vulnerability of the brain to amyloid-beta amyloidogenesis, is essential for cognitive, emotional, and synaptic functions. J Neurosci. 2005; 25(50):11693-709. PMC: 2564291. DOI: 10.1523/JNEUROSCI.2766-05.2005. View

Thery C, Amigorena S, Raposo G, Clayton A . Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol. 2008; Chapter 3:Unit 3.22. DOI: 10.1002/0471143030.cb0322s30. View

Simhadri V, Reiners K, Hansen H, Topolar D, Simhadri V, Nohroudi K . Dendritic cells release HLA-B-associated transcript-3 positive exosomes to regulate natural killer function. PLoS One. 2008; 3(10):e3377. PMC: 2566590. DOI: 10.1371/journal.pone.0003377. View

Seow Y, Wood M . Biological gene delivery vehicles: beyond viral vectors. Mol Ther. 2009; 17(5):767-77. PMC: 2835126. DOI: 10.1038/mt.2009.41. View

Yin H, Moulton H, Betts C, Seow Y, Boutilier J, Iverson P . A fusion peptide directs enhanced systemic dystrophin exon skipping and functional restoration in dystrophin-deficient mdx mice. Hum Mol Genet. 2009; 18(22):4405-14. DOI: 10.1093/hmg/ddp395. View

Schorey J, Bhatnagar S . Exosome function: from tumor immunology to pathogen biology. Traffic. 2008; 9(6):871-81. PMC: 3636814. DOI: 10.1111/j.1600-0854.2008.00734.x. View

Nunes F, Furth E, Wilson J, Raper S . Gene transfer into the liver of nonhuman primates with E1-deleted recombinant adenoviral vectors: safety of readministration. Hum Gene Ther. 1999; 10(15):2515-26. DOI: 10.1089/10430349950016852. View

10.

Kumar P, Wu H, McBride J, Jung K, Kim M, Davidson B . Transvascular delivery of small interfering RNA to the central nervous system. Nature. 2007; 448(7149):39-43. DOI: 10.1038/nature05901. View

11.

Nishitomi K, Sakaguchi G, Horikoshi Y, Gray A, Maeda M, Hirata-Fukae C . BACE1 inhibition reduces endogenous Abeta and alters APP processing in wild-type mice. J Neurochem. 2006; 99(6):1555-63. DOI: 10.1111/j.1471-4159.2006.04178.x. View

12.

Flint P, Bo Li Z, Lehar M, Saito K, Pai S . Laryngeal muscle surface receptors identified using random phage library. Laryngoscope. 2005; 115(11):1930-7. DOI: 10.1097/01.mlg.0000172273.98418.8d. View

13.

Quah B, ONeill H . The immunogenicity of dendritic cell-derived exosomes. Blood Cells Mol Dis. 2005; 35(2):94-110. DOI: 10.1016/j.bcmd.2005.05.002. View

14.

Simpson R, Jensen S, Lim J . Proteomic profiling of exosomes: current perspectives. Proteomics. 2008; 8(19):4083-99. DOI: 10.1002/pmic.200800109. View

15.

Skog J, Wurdinger T, van Rijn S, Meijer D, Gainche L, Sena-Esteves M . Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat Cell Biol. 2008; 10(12):1470-6. PMC: 3423894. DOI: 10.1038/ncb1800. View

16.

Gonzalez S, McMenamin M, Charlton H, Goodman J, Lantos T, Simpson C . Readministration of adenoviral gene delivery to dopamine neurons. Neuroreport. 2007; 18(15):1609-14. DOI: 10.1097/WNR.0b013e3282f03fe5. View

17.

Kao S, Krichevsky A, Kosik K, Tsai L . BACE1 suppression by RNA interference in primary cortical neurons. J Biol Chem. 2003; 279(3):1942-9. DOI: 10.1074/jbc.M309219200. View

18.

Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S . Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med. 1992; 176(6):1693-702. PMC: 2119469. DOI: 10.1084/jem.176.6.1693. View

19.

Mosmann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2):55-63. DOI: 10.1016/0022-1759(83)90303-4. View

20.

Larson S, Jackson L, Chen L, Rychahou P, Evers B . Effectiveness of siRNA uptake in target tissues by various delivery methods. Surgery. 2007; 142(2):262-9. PMC: 2427156. DOI: 10.1016/j.surg.2007.03.011. View