FUS-mediated HypEVs: Neuroprotective effects against ischemic stroke

Yousheng Wu; Xiaoxiong Huang; Zefeng Tan; Jiankun Zang; Min Peng; Niu He; Tao Zhang; Hongcheng Mai; Anding Xu; Dan Lu

doi:10.1016/j.bioactmat.2023.07.009

FUS-mediated HypEVs: Neuroprotective effects against ischemic stroke

Bioact Mater. 2023 Jul 16:29:196-213. doi: 10.1016/j.bioactmat.2023.07.009. eCollection 2023 Nov.

Authors

Yousheng Wu^{1

2

3}, Xiaoxiong Huang^{1

2

4}, Zefeng Tan⁵, Jiankun Zang^{1

2

3}, Min Peng^{1

2

3}, Niu He^{1

3}, Tao Zhang⁶, Hongcheng Mai^{1

2

7

8}, Anding Xu^{1

2

3}, Dan Lu^{1

2

3}

Affiliations

¹ Department of Neurology and Stroke Center, The First Affiliated Hospital of Jinan University, Guangzhou, China.
² Clinical Neuroscience Institute, The First Affiliated Hospital of Jinan University, Guangzhou, China.
³ Key Lab of Guangzhou Basic and Translational Research of Pan-vascular Diseases, The First Affiliated Hospital of Jinan University, Guangzhou, China.
⁴ Department of Neurology and Stroke Center, The Central Hospital of Shaoyang, Hunan, China.
⁵ Department of Neurology, The First People's Hospital of Foshan, Guangdong, China.
⁶ Department of Cardiology, The First Affiliated Hospital of Jinan University, Guangzhou, China.
⁷ Munich Medical Research School (MMRS), Ludwig-Maximilians University Munich, Munich, Germany.
⁸ Insititute for Tissue Engineering and Regenerative Medicine (iTERM), Helmholtz Zentrum München, Neuherberg, Germany.

Abstract

Few studies have investigated the properties and protein composition of small extracellular vesicles (sEVs) derived from neurons under hypoxic conditions. Presently, the extent of the involvement of these plentiful sEVs in the onset and progression of ischemic stroke remains an unresolved question. Our study systematically identified the characteristics of sEVs derived from neurons under hypoxic conditions (HypEVs) by physical characterization, sEV absorption, proteomics and transcriptomics analysis. The effects of HypEVs on neurites, cell survival, and neuron structure were assessed in vitro and in vivo by neural complexity tests, magnetic resonance imaging (MRI), Golgi staining, and Western blotting of synaptic plasticity-related proteins and apoptotic proteins. Knockdown of Fused in Sarcoma (FUS) small interfering RNA (siRNA) was used to validate FUS-mediated HypEV neuroprotection and mitochondrial mRNA release. Hypoxia promoted the secretion of sEVs, and HypEVs were more easily taken up and utilized by recipient cells. The MRI results illustrated that the cerebral infarction volume was reduced by 45% with the application of HypEVs, in comparison to the non- HypEV treatment group. Mechanistically, the FUS protein is necessary for the uptake and neuroprotection of HypEVs against ischemic stroke as well as carrying a large amount of mitochondrial mRNA in HypEVs. However, FUS knockdown attenuated the neuroprotective rescue capabilities of HypEVs. Our comprehensive dataset clearly illustrates that FUS-mediated HypEVs deliver exceptional neuroprotective effects against ischemic stroke, primarily through the maintenance of neurite integrity and the reduction of mitochondria-associated apoptosis.

Keywords: FUS; Hypoxia; Mitochondrial mRNA; Neuron-derived small extracellular vesicle; Neuroprotection.