Skip to main content

Advertisement

SpringerLink
Log in

Evaluation of lncRNA Expression During the Differentiation of Mesenchymal Stem Cells to Insulin-Secreting Progenitors

  • Published:
Molecular Neurobiology Aims and scope Submit manuscript

Abstract

Diabetes mellitus is a metabolic disease caused by a defect in insulin secretion, insulin function, or both that destroys pancreatic islet beta cells. There is ample evidence that long non-coding RNAs (lncRNAs) play a vital role in cell formation and differentiation. The present study aims to investigate the expression pattern of specific lncRNAs in mesenchymal stem cell (MSC) differentiation into insulin-producing beta cell (IPCs) progenitors for cell therapy purposes. MSCs were extracted from human umbilical cord Wharton jelly (hWJ-MSCs) using the explant method and cultured in two-dimensional (2D) and three-dimensional (3D) media on polylactic acid/Wax (PLA/Wax) nanofibrous scaffold using a three-step protocol containing CHIR99021 small molecules and Indolactam V. At the end of each differentiation step, immunocytochemistry and qRT-PCR were used to confirm the differentiation at the protein and RNA levels and the expression changes of six selective lncRNAs were evaluated by qRT-PCR. The results indicated that the expression of the selected lncRNAs was significantly altered during the differentiation process into beta progenitor cells, indicating their potential role in regulating the IPC differentiation process. More specifically, all of the desired lncRNAs demonstrated a significant increase during the beta cell differentiation, with HI-LNC71 and HI-LNA12 experiencing the highest expression in the produced Beta cell progenitors respectively (p<0.0001). These results can be valuable in tissue engineering and treatment studies by replacing beta precursor cells to control diabetic patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data Availability

Available upon request.

References

  1. Hoveizi E, Mohammadi T (2019) Differentiation of endometrial stem cells into insulin-producing cells using signaling molecules and zinc oxide nanoparticles, and three-dimensional culture on nanofibrous scaffolds. J Mater Sci Mater Med 30(9):101. https://doi.org/10.1007/s10856-019-6301-3

    Article  CAS  PubMed  Google Scholar 

  2. Gooraninejad S, Hoveizi E, Hushmandi K, Gooraninejad S, Tabatabaei SRF (2020) Small molecule differentiate PDX1-expressing cells derived from human endometrial stem cells on PAN electrospun nanofibrous scaffold: applications for the treatment of diabetes in rat. Mol Neurobiol 57:3969–3978

    Article  CAS  PubMed  Google Scholar 

  3. Zeka K, Alfa HH, Ruparelia KC, Arroo RR (2019) Use of natural products in the prevention and management of type 2 diabetes. Stud Nat Prod Chem 63:197–210

    Article  CAS  Google Scholar 

  4. Abels M, Riva M, Shcherbina L, Fischer A-HT, Banke E, Degerman E, Lindqvist A, Wierup N (2022) Overexpressed beta cell CART increases insulin secretion in mouse models of insulin resistance and diabetes. Peptides 151:170747

    Article  CAS  PubMed  Google Scholar 

  5. Memon B, Abdelalim EM (2020) Stem cell therapy for diabetes: beta cells versus pancreatic progenitors. Cells 9(2):283

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Zhou Z, Zhu X, Huang H, Xu Z, Jiang J, Chen B, Zhu H (2022) Recent progress of research regarding the applications of stem cells for treating diabetes mellitus. Stem Cells Dev 31(5-6):102–110

    Article  CAS  PubMed  Google Scholar 

  7. Elham H, Mahmoud H (2018) The effect of pancreas islet-releasing factors on the direction of embryonic stem cells towards Pdx1 expressing cells. Appl Biochem Biotechnol. https://doi.org/10.1007/s12010-018-2733-3

  8. Hoveizi E, Tavakol S, Shirian S, Sanamiri K (2019) Electrospun nanofibers for diabetes: tissue engineering and cell-based therapies. Curr Stem Cell Res Ther 14(2):152–168

    Article  CAS  PubMed  Google Scholar 

  9. Solis MA, Moreno Velásquez I, Correa R, Huang LL (2019) Stem cells as a potential therapy for diabetes mellitus: a call-to-action in Latin America. Diabetol Metab Syndr 11(1):1–13

    Article  Google Scholar 

  10. Rogal J, Zbinden A, Schenke-Layland K, Loskill P (2019) Stem-cell based organ-on-a-chip models for diabetes research. Adv Drug Deliv Rev 140:101–128

    Article  CAS  PubMed  Google Scholar 

  11. Sneddon JB, Tang Q, Stock P, Bluestone JA, Roy S, Desai T, Hebrok M (2018) Stem cell therapies for treating diabetes: progress and remaining challenges. Cell Stem Cell 22(6):810–823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Motterle A, Gattesco S, Caille D, Meda P, Regazzi R (2015) Involvement of long non-coding RNAs in beta cell failure at the onset of type 1 diabetes in NOD mice. Diabetologia 58(8):1827–1835

    Article  CAS  PubMed  Google Scholar 

  13. Long Y, Wang X, Youmans DT, Cech TR (2017) How do lncRNAs regulate transcription? Sci Adv 3(9):eaao2110

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ismail N, Abdullah N, Abdul Murad NA, Jamal R, Sulaiman SA (2021) Long non-coding RNAs (lncRNAs) in cardiovascular disease complication of type 2 diabetes. Diagnostics 11(1):145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Font-Cunill B, Arnes L, Ferrer J, Sussel L, Beucher A (2018) Long non-coding RNAs as local regulators of pancreatic islet transcription factor genes. Front Genet 524

  16. Wong WK, Sørensen AE, Joglekar MV, Hardikar AA, Dalgaard LT (2018) Non-coding RNA in pancreas and β-cell development. Non-coding RNA 4(4):41

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Morán I, Akerman I, Van De Bunt M, Xie R, Benazra M, Nammo T, Arnes L, Nakić N et al (2012) Human β cell transcriptome analysis uncovers lncRNAs that are tissue-specific, dynamically regulated, and abnormally expressed in type 2 diabetes. Cell Metab 16(4):435–448

    Article  PubMed  PubMed Central  Google Scholar 

  18. Akerman I, Tu Z, Beucher A, Rolando DM, Sauty-Colace C, Benazra M, Nakic N, Yang J et al (2017) Human pancreatic β cell lncRNAs control cell-specific regulatory networks. Cell Metab 25(2):400–411

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Guo J, Liu Z, Gong R (2019) Long noncoding RNA: an emerging player in diabetes and diabetic kidney disease. Clin Sci 133(12):1321–1339

    Article  CAS  Google Scholar 

  20. Dieter C, Lemos NE, Corrêa NRDF, Assmann TS, Crispim D (2021) The impact of lncRNAs in diabetes mellitus: a systematic review and in silico analyses. Front Endocrinol 12:187

    Article  Google Scholar 

  21. Chen Y, He Y, Zhou H (2020) The potential role of lncRNAs in diabetes and diabetic microvascular complications. Endocr J 67:EJ19-0574

    Article  Google Scholar 

  22. Paladino FV, Peixoto-Cruz JS, Santacruz-Perez C, Goldberg AC (2016) Comparison between isolation protocols highlights intrinsic variability of human umbilical cord mesenchymal cells. Cell Tissue Bank 17(1):123–136

    Article  CAS  PubMed  Google Scholar 

  23. Corritore E, Dugnani E, Pasquale V, Misawa R, Witkowski P, Lei J, Markmann J, Piemonti L et al (2014) β-cell differentiation of human pancreatic duct–derived cells after in vitro expansion. Cell Reprogram (Formerly “Cloning and Stem Cells”) 16(6):456–466

    CAS  Google Scholar 

  24. Kim D-W, Staples M, Shinozuka K, Pantcheva P, Kang S-D, Borlongan CV (2013) Wharton’s jelly-derived mesenchymal stem cells: phenotypic characterization and optimizing their therapeutic potential for clinical applications. Int J Mol Sci 14(6):11692–11712

    Article  PubMed  PubMed Central  Google Scholar 

  25. Mimura I, Nangaku M (2023) Epigenetic regulation of angiogenesis and ischemic response by long noncoding RNA LEENE in diabetes. Kidney Int 23:00483-0

    Google Scholar 

  26. Lim JE, Hong K-W, Jin H-S, Kim YS, Park HK, Oh B (2010) Type 2 diabetes genetic association database manually curated for the study design and odds ratio. BMC Med Inform Decis Mak 10(1):1–12

    Article  Google Scholar 

  27. Arnes L, Akerman I, Balderes DA, Ferrer J, Sussel L (2016) βlinc1 encodes a long noncoding RNA that regulates islet β-cell formation and function. Genes Dev 30(5):502–507

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Khoo C, Yang J, Weinrott SA, Kaestner KH, Naji A, Schug J, Stoffers DA (2012) Research resource: the pdx1 cistrome of pancreatic islets. Mol Endocrinol 26(3):521–533

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Kameswaran V, Bramswig NC, McKenna LB, Penn M, Schug J, Hand NJ, Chen Y, Choi I et al (2014) Epigenetic regulation of the DLK1-MEG3 microRNA cluster in human type 2 diabetic islets. Cell Metab 19(1):135–145

    Article  CAS  PubMed  Google Scholar 

  30. Shahjalal HM, Shiraki N, Sakano D, Kikawa K, Ogaki S, Baba H, Kume K, Kume S (2014) Generation of insulin-producing β-like cells from human iPS cells in a defined and completely xeno-free culture system. J Mol Cell Biol 6(5):394–408

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Shahid Chamran University of Ahvaz for funding this study.

Funding

This study was funded and supported by the Shahid Chamran University of Ahvaz (GrantNumber:SCU.SB1401.12464).

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Tina Shafaf, Sayed Reza Kazeminejad & Elham Hoveizi

Authors
  1. Tina Shafaf
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sayed Reza Kazeminejad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elham Hoveizi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

T. Sh. carried out the experiment and wrote the manuscript. SR. K. contributed to data and statistical analysis and interpretation of data. E. H. designed and directed the project. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Elham Hoveizi.

Ethics declarations

Ethics Approval

The animal experiment was approved by the Animal Ethics Committee of Shahid Chamran University (Approval No. EE/99.3.02.65844/scu.ac.ir).

Consent to Participate

Written informed consent was obtained for all human patient samples.

Consent for Publication

Not applicable.

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shafaf, T., Kazeminejad, S.R. & Hoveizi, E. Evaluation of lncRNA Expression During the Differentiation of Mesenchymal Stem Cells to Insulin-Secreting Progenitors. Mol Neurobiol 61, 372–384 (2024). https://doi.org/10.1007/s12035-023-03571-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12035-023-03571-w

Keywords

Search

Navigation