Central nervous system(CNS)injuries,including stroke,traumatic brain injury,and spinal cord injury,are essential causes of death and long-term disability and are difficult to cure,mainly due to the limited neuron rege...Central nervous system(CNS)injuries,including stroke,traumatic brain injury,and spinal cord injury,are essential causes of death and long-term disability and are difficult to cure,mainly due to the limited neuron regeneration and the glial scar formation.Herein,we apply extracellular vesicles(EVs)secreted by M2 microglia to improve the differentiation of neural stem cells(NSCs)at the injured site,and simultaneously modify them with the injured vascular targeting peptide(DA7R)and the stem cell recruiting factor(SDF-1)on their surface via copper-free click chemistry to recruit NSCs,inducing their neuronal differentiation,and serving as the nanocarriers at the injured site(Dual-EV).Results prove that the Dual-EV could target human umbilical vascular endothelial cells(HUVECs),recruit NSCs,and promote the neuronal differentiation of NSCs in vitro.Furthermore,10 miRNAs are found to be upregulated in Dual-M2-EVs compared to Dual-M0-EVs via bioinformatic analysis,and further NSC differentiation experiment by flow cytometry reveals that among these miRNAs,miR30b-3p,miR-222-3p,miR-129-5p,and miR-155-5p may exert effect of inducing NSC to differentiate into neurons.In vivo experiments show that Dual-EV nanocarriers achieve improved accumulation in the ischemic area of stroke model mice,potentiate NSCs recruitment,and increase neurogenesis.This work provides new insights for the treatment of neuronal regeneration after CNS injuries as well as endogenous stem cells,and the click chemistry EV/peptide/chemokine and related nanocarriers for improving human health.展开更多
Background Healthy plasma therapy reverses cognitive deficits and promotes neuroplasticity in ageing brain disease.However,whether healthy plasma therapy improve blood-brain barrier integrity after stroke remains unkn...Background Healthy plasma therapy reverses cognitive deficits and promotes neuroplasticity in ageing brain disease.However,whether healthy plasma therapy improve blood-brain barrier integrity after stroke remains unknown.Methods Here,we intravenously injected healthy female mouse plasma into adult female ischaemic stroke C57BL/6 mouse induced by 90 min transient middle cerebral artery occlusion for eight consecutive days.Infarct volume,brain atrophy and neurobehavioural tests were examined to assess the outcomes of plasma treatment.Cell apoptosis,blood-brain barrier integrity and fibroblast growth factor 21 knockout mice were used to explore the underlying mechanism.Results Plasma injection improved neurobehavioural recovery and decreased infarct volume,brain oedema and atrophy after stroke.Immunostaining showed that the number of transferase dUTP nick end labelling+/NeuN+cells decreased in the plasma-injected group.Meanwhile,plasma injection reduced ZO-1,occluding and claudin-5 tight junction gap formation and IgG extravasation at 3 days after ischaemic stroke.Western blot results showed that the FGF21 expression increased in the plasma-injected mice.However,using FGF21 knockout mouse plasma injecting to the ischaemic wild-type mice diminished the neuroprotective effects.Conclusions Our study demonstrated that healthy adult plasma treatment protected the structural and functional integrity of blood-brain barrier,reduced neuronal apoptosis and improved functional recovery via FGF21,opening a new avenue for ischaemic stroke therapy.展开更多
基金support from Harvard/MITsupport by grants from the National Key R&D Program of China(2019YFA0112000)+5 种基金National Natural Science Foundation of China(Nos.81930051,82003658,81801170)the Interdisciplinary Program of Shanghai Jiao Tong University(ZH2018ZDA04,China)Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support(20171906,China)China Postdoctoral Science Foundation(2019M661546)National Postdoctoral Program for Innovative Talents(BX20200212,China)Zhejiang Provincial Natural Science Foundation of China(No.LQ21H300009,China)。
文摘Central nervous system(CNS)injuries,including stroke,traumatic brain injury,and spinal cord injury,are essential causes of death and long-term disability and are difficult to cure,mainly due to the limited neuron regeneration and the glial scar formation.Herein,we apply extracellular vesicles(EVs)secreted by M2 microglia to improve the differentiation of neural stem cells(NSCs)at the injured site,and simultaneously modify them with the injured vascular targeting peptide(DA7R)and the stem cell recruiting factor(SDF-1)on their surface via copper-free click chemistry to recruit NSCs,inducing their neuronal differentiation,and serving as the nanocarriers at the injured site(Dual-EV).Results prove that the Dual-EV could target human umbilical vascular endothelial cells(HUVECs),recruit NSCs,and promote the neuronal differentiation of NSCs in vitro.Furthermore,10 miRNAs are found to be upregulated in Dual-M2-EVs compared to Dual-M0-EVs via bioinformatic analysis,and further NSC differentiation experiment by flow cytometry reveals that among these miRNAs,miR30b-3p,miR-222-3p,miR-129-5p,and miR-155-5p may exert effect of inducing NSC to differentiate into neurons.In vivo experiments show that Dual-EV nanocarriers achieve improved accumulation in the ischemic area of stroke model mice,potentiate NSCs recruitment,and increase neurogenesis.This work provides new insights for the treatment of neuronal regeneration after CNS injuries as well as endogenous stem cells,and the click chemistry EV/peptide/chemokine and related nanocarriers for improving human health.
基金This study was supported by grants from the Scientific Research and Innovation Program of Shanghai Education Commission 2019-01-07-00-02-E00064(G-YY)National Key R&D,Program of China#2016YFC1300602(G-YY),#2019YFA0112000(YT)+1 种基金the National Natural Science Foundation of China(NSFC)projects 81771251(G-YY),81801170(YT),82071284(YT),81771244(ZZ),81974179(ZZ),81870921(YW)K.C.Wong Education Foundation(G-YY,no award/grant number).
文摘Background Healthy plasma therapy reverses cognitive deficits and promotes neuroplasticity in ageing brain disease.However,whether healthy plasma therapy improve blood-brain barrier integrity after stroke remains unknown.Methods Here,we intravenously injected healthy female mouse plasma into adult female ischaemic stroke C57BL/6 mouse induced by 90 min transient middle cerebral artery occlusion for eight consecutive days.Infarct volume,brain atrophy and neurobehavioural tests were examined to assess the outcomes of plasma treatment.Cell apoptosis,blood-brain barrier integrity and fibroblast growth factor 21 knockout mice were used to explore the underlying mechanism.Results Plasma injection improved neurobehavioural recovery and decreased infarct volume,brain oedema and atrophy after stroke.Immunostaining showed that the number of transferase dUTP nick end labelling+/NeuN+cells decreased in the plasma-injected group.Meanwhile,plasma injection reduced ZO-1,occluding and claudin-5 tight junction gap formation and IgG extravasation at 3 days after ischaemic stroke.Western blot results showed that the FGF21 expression increased in the plasma-injected mice.However,using FGF21 knockout mouse plasma injecting to the ischaemic wild-type mice diminished the neuroprotective effects.Conclusions Our study demonstrated that healthy adult plasma treatment protected the structural and functional integrity of blood-brain barrier,reduced neuronal apoptosis and improved functional recovery via FGF21,opening a new avenue for ischaemic stroke therapy.