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Using Human Fetal Neural Stem Cells to Elucidate the Role of the JAK-STAT Cell Signaling Pathway in Oligodendrocyte Differentiation In Vitro

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Abstract

Oligodendrocytes (OL) are the myelinating cells of the central nervous system that mediate nerve conduction. Loss of oligodendrocytes results in demyelination, triggering neurological deficits. Develo** a better understanding of the cell signaling pathways influencing OL development may aid in the development of therapeutic strategies. The primary focus of this study was to investigate and elucidate the cell signaling pathways implicated in the developmental maturation of oligodendrocytes using human fetal neural stem cells (hFNSCs)–derived primary OL and MO3.13 cell line. Successful differentiation into OL was established by examining morphological changes, increased expression of mature OL markers MBP, MOG and decreased expression of pre-OL markers CSPG4 and O4. Analyzing transcriptional datasets (using RNA sequencing) in pre-OL and mature OL derived from hFNSCs revealed the novel and critical involvement of the JAK-STAT cell signaling pathway in terminal OL maturation. The finding was validated in MO3.13 cell line whose differentiation was accompanied by upregulation of IL-6 and the transcription factor STAT3. Increased phosphorylated STAT3 (pY705) levels were demonstrated by western blotting in hFNSCs-derived primary OL as well as terminal maturation in MO3.13 cells, thus validating the involvement of the JAK-STAT pathway in OL maturation. Pharmacological suppression of STAT3 phosphorylation (confirmed by western blotting) was able to prevent the increase of MBP-positive cells as demonstrated by flow cytometry. These novel findings highlight the involvement of the JAK-STAT pathway in OL maturation and raise the possibility of using this as a therapeutic strategy in demyelinating diseases.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors wish to acknowledge the support of the facilities provided under the Biotechnology Information System Network (BTISNET) grant, DBT, Govt. of India, and Distributed Information Centre at NBRC, Manesar, India.

Funding

This study received financial support from the Department of Biotechnology (DBT), Govt. of India—extramural research grant (BT/PR21413/MED/122/40/2016).

Author information

Author notes

  1. Devanjan Dey and Sagar Tyagi have contributed equally.

Authors and Affiliations

  1. Department of Biochemistry, All India Institute of Medical Sciences, Room No. 3027A, New Delhi, 110029, India

    Devanjan Dey, Sagar Tyagi, Vadanya Shrivastava, Sweety Rani, Subrata Sinha, Jayanth Kumar Palanichamy & Sudip Sen

  2. Department of Obstetrics and Gynaecology, All India Institute of Medical Sciences, New Delhi, India

    Jai Bhagwan Sharma

  3. Department of Molecular and Cellular Neuroscience, National Brain Research Centre, Manesar, Haryana, India

    Pankaj Seth

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Contributions

Conception and design of work: Devanjan Dey, Sagar Tyagi, Sudip Sen, and Pankaj Seth. Data collection: Devanjan Dey, Sagar Tyagi, Vadanya Shrivastava, and Jai Bhagwan Sharma. Data analysis and interpretation: Devanjan Dey, Sagar Tyagi, Vadanya Shrivastava, and Sudip Sen. Drafting article: Devanjan Dey, Sagar Tyagi, Vadanya Shrivastava, and Sudip Sen. Critical revision of article: Sudip Sen, Jayanth Kumar Palanichamy, Subrata Sinha, Pankaj Seth, and Jai Bhagwan Sharma. Final approval of the version to be published: all.

Corresponding author

Correspondence to Sudip Sen.

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Ethics Approval

This study was performed in line with the principles of the Declaration of Helsinki. Necessary approval was taken from the Institutional Ethics Committee (IEC-180/07.04.2017 dated 12 April 2017) and Institutional Committee for Stem Cell Research (IC-SCR/66/17(O) dated 18 September 2017), All India Institute of Medical Sciences, New Delhi, prior to commencement of work.

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Informed consent was obtained from mothers undergoing medical termination of pregnancy (MTP).

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The authors declare no competing interests.

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Dey, D., Tyagi, S., Shrivastava, V. et al. Using Human Fetal Neural Stem Cells to Elucidate the Role of the JAK-STAT Cell Signaling Pathway in Oligodendrocyte Differentiation In Vitro. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-03928-9

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