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Overexpression of SMN2 Gene in Motoneuron-Like Cells Differentiated from Adipose-Derived Mesenchymal Stem Cells by Ponasterone A

  • Rashin Mohseni
  • Mahmood Reza Ashrafi
  • Jafar Ai
  • Mahin Nikougoftar
  • Mahmoud Mohammadi
  • Masood Ghahvechi-Akbari
  • Alireza Shoae-Hassani
  • Amir Ali Hamidieh
Article
  • 7 Downloads

Abstract

Cell therapy and stem cell transplantation strategies have provided potential therapeutic approaches for the treatment of neurological disorders. Adipose-derived mesenchymal stem cells (ADMSCs) are abundant adult stem cells with low immunogenicity, which can be used for allogeneic cell replacement therapies. Differentiation of ADMSCs into acetylcholine-secreting motoneurons (MNs) is a promising treatment for MN diseases, such as spinal muscular atrophy (SMA), which is associated with the level of SMN1 gene expression. The SMN2 gene plays an important role in MN disorders, as it can somewhat compensate for the lack of SMN1 expression in SMA patients. Although the differentiation potential of ADMSCs into MNs has been previously established, overexpression of SMN2 gene in a shorter period with a longer survival has yet to be elucidated. Ponasterone A (PNA), an ecdysteroid hormone activating the PI3K/Akt pathway, was studied as a new steroid to promote SMN2 overexpression in MNs differentiated from ADMSCs. After induction with retinoic acid, sonic hedgehog, forskolin, and PNA, MN phenotypes were differentiated from ADMSCs, and immunochemical staining, specific for β-tubulin, neuron-specific enolase, and choline acetyltransferase, was performed. Also, the results of real-time PCR assay indicated nestin, Pax6, Nkx2.2, Hb9, Olig2, and SMN2 expression in the differentiated cells. After 2 weeks of treatment, cultures supplemented with PNA showed a longer survival and a 1.2-fold increase in the expression of SMN2 (an overall 5.6-fold increase; *P ≤ 0.05), as confirmed by the Western blot analysis. The PNA treatment increased the levels of ChAT, Isl1, Hb9, and Nkx2 expression in MN-like cells. Our findings highlight the role of PNA in the upregulation of SMN2 genes from MSC-derived MN-like cells, which may serve as a potential candidate in cellular therapy for SMA patients.

Keywords

Mesenchymal stem cell Neurology Motoneuron SMN Werdnig-Hoffmann disease 

Notes

Compliance with Ethical Standards

The fat tissue was obtained after proper consenting process and approvals from Tehran University of Medical Sciences ethics committee.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Rashin Mohseni
    • 1
  • Mahmood Reza Ashrafi
    • 2
  • Jafar Ai
    • 1
  • Mahin Nikougoftar
    • 3
  • Mahmoud Mohammadi
    • 2
  • Masood Ghahvechi-Akbari
    • 2
  • Alireza Shoae-Hassani
    • 1
  • Amir Ali Hamidieh
    • 1
    • 4
  1. 1.Applied Cell Sciences and Tissue Engineering Department, School of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran
  2. 2.Pediatric Neurology Division, Children’s Medical Center, Pediatric Center of ExcellenceTehran University of Medical SciencesTehranIran
  3. 3.Blood Transfusion Research Center, High Institute for Research and Education in Transfusion medicine, Iranian Blood Transfusion Organization (IBTO)TehranIran
  4. 4.Pediatric Hematology, Oncology and Stem Cell Transplantation Department, Children’s Medical Center, Pediatric Center of ExcellenceTehran University of Medical SciencesTehranIran

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