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Let-7a-5p regulated by lncRNA-MEG3 promotes functional differentiation to Schwann cells from adipose derived stem cells via directly inhibiting RBPJ-mediating Notch pathway

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Abstract

Schwann cells (SCs) have important roles in supporting and repairing peripheral neurons, and thus have great potential for nerve injury treatment. Adipose tissue-derived stem cells (ADSCs) can be reliably induced to differentiate into SCs. However, the underlying molecular mechanisms are unclear. We explored the roles of MEG3/let-7a-5p/RBPJ axis in the differentiation into SCs from ADSCs. Primary ADSCs were induced to differentiate into SCs by appropriate reagents. ELISA, immunostaining, Western blotting, and qRT-PCR were employed to examine levels of SC-markers such as S100, GFAP, SOX10, p75NTR, GAP43, MPZ, β-NGF, BDNF, and NCAM and let-7 family, MEG3, RBPJ, and Notch signaling related proteins. Dual luciferase assay and RNA immunoprecipitation were performed to validate interactions of let-7a-5p/RBPJ mRNA and MEG3/let-7a-5p. Cultured ADSCs could be induced to differentiate into functional SCs. Let-7a-5p and let-7d-5p were elevated during the differentiation while MEG3 and RBPJ/Notch-signaling were suppressed. Let-7a-5p mimics promoted ADSC differentiation into SCs and up-regulated the levels of SC-related markers including S100, GFAP, SOX10, p75NTR, GAP43, MPZ, β-NGF, and NCAM, while RBPJ or MEG3 overexpression retarded the differentiation and reduced those levels. Let-7a-5p directly targeted RBPJ and MEG3 disinhibited Notch-RBPJ signaling via sponging let-7a-5p. RBPJ overexpression reversed the acceleration of let-7a-5p mimics on SC differentiation while let-7a-5p mimics blocked MEG3-mediated suppression on SC differentiation. Let-7a-5p sponged by MEG3 promotes differentiation of ADSCs into SCs via suppressing Notch signaling by targeting RBPJ. These findings shed light on mechanisms underlying the differentiation of ADSCs to SCs and provide avenues to accelerate the process.

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Data availability

All data generated or analyzed during this study are included in this published article.

Code availability

Not applicable.

Abbreviations

SC:

Schwann cell

NSC:

Neural stem cell

ADSC:

Adipose tissue-derived stem cell

NICD:

Notch intracellular domain

RBPJ:

Recombination signal-binding protein for immunoglobulin κ J region

BMSC:

Bone marrow mesenchymal stem cell

ncRNA:

Non-coding RNAs

miRNA:

Micro RNA

lncRNA:

Long non-coding RNA

ceRNA:

Competing endogenous RNA

MEG3:

Maternally expressed gene 3

DMEM:

Dulbecco's Modified Eagle Medium

FBS:

Fetal bovine serum

PDGF:

Platelet-derived growth factor

FFGF:

Fibroblast Growth Factor

β-NGF:

β-Nerve growth factor

BDNF:

Brain-derived neurotrophic factor

IL-6:

Interleukin 6

NCAM:

Neural cell adhesion molecule

GFAP:

Glial fibrillary acidic protein

HES1/5:

Hairy and enhancer of split 1/5

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Funding

This work was supported by National Natural Science Foundation of China (Grant/Award Number: 81601083, 81702238, 81401006).

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Authors and Affiliations

  1. Department of Neurology, The Second Xiangya Hospital, Central South University, Changsha, 410011, Hunan, China

    Wei Wang, Mei-Feng Gu, Xiang-Min Shen & Jie Zhang

  2. Department of Neurosurgery, The Third Xiangya Hospital of Central South University, No. 138, Tongzipo Road, Changsha, 410078, Hunan, China

    Zhi-Fei Wang & Liang Yang

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  1. Wei Wang
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  2. Mei-Feng Gu
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Correspondence to Liang Yang.

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Wang, W., Gu, MF., Wang, ZF. et al. Let-7a-5p regulated by lncRNA-MEG3 promotes functional differentiation to Schwann cells from adipose derived stem cells via directly inhibiting RBPJ-mediating Notch pathway. Apoptosis 26, 548–560 (2021). https://doi.org/10.1007/s10495-021-01685-x

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