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Human urine-derived stem cell-derived exosomal miR-21-5p promotes neurogenesis to attenuate Rett syndrome via the EPha4/TEK axis


Rett syndrome (RTT) is a rare neurodevelopmental disorder that results in multiple disabilities. Exosomal microRNA (miRs) from urine-derived stem cells (USCs) have been shown to induce neurogenesis and aid in functional recovery from brain ischemia. In the present study, we sought to determine whether that exosomal miR-21-5p from USCs could promote early neural formation in a model of RTT. USCs were isolated and evaluated by flow cytometry. Exosomes were analyzed by transmission electron microscopy, tunable resistive pulse sensing (TRPS), and western blotting. PKH26 fluorescent dyes were used to observe intake of exosomes in vivo and in vitro. An RTT mouse model was treated with exosomes for behavioral studies. Dual‐luciferase report gene assays were conducted to evaluate the relationship between miR-21-5p and Eph receptor A4 (EphA4). In vitro, treatment with exosomes from human urine‐derived stem cells (USC-Exos) increased the percentage of neuron-specific class III beta-tubulin (Tuj1)+ nerve cells as well as the transcription levels of β-III tubulin and doublecortin (DCX). A higher level of miR-21-5p was observed in USC-Exos, which promoted differentiation in NSCs by targeting the EPha4/TEK axis. In vivo, exosomal miR-21-5p improved the behavior, motor coordination, and cognitive ability of mice, facilitated the differentiation of NSCs in the subventricular zone of the lateral ventricle and promoted a marked rise in the number of DCX+ cells. Our data provide evidence that exosomal miR-21-5p from human USCs facilitate early nerve formation by regulating the EPha4/TEK axis.

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Fig. 1: The identification of USCs and USC-Exos.
Fig. 2: USC-Exos promoted neuronal differentiation of RTT NSCs.
Fig. 3: USC-Exos promoted neural differentiation of RTT NSCs by inhibiting the expression of EPha4.
Fig. 4: USC-Exos mediated miR-21-5p to promote neuronal differentiation of RTT NSCs by targeting EPha4.
Fig. 5: Exosomal miR-21-5p promoted neurogenesis in RTT mice.
Fig. 6: Exosomal miR-21-5p promoted neurogenesis in RTT mice.
Fig. 7: Mechanism diagram.


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The authors would like to acknowledge the helpful comments on this paper received from the reviewers.

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WP designed the study. WP, XHX, and MZ collated the data, carried out data analyses, and produced the initial draft of the manuscript. XYS contributed to drafting the manuscript. All authors have read and approved the final submitted manuscript.

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Correspondence to Xingyu Song.

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Pan, W., Xu, X., Zhang, M. et al. Human urine-derived stem cell-derived exosomal miR-21-5p promotes neurogenesis to attenuate Rett syndrome via the EPha4/TEK axis. Lab Invest 101, 824–836 (2021).

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