Antisense miR-7 impairs insulin expression in developing pancreas and in cultured pancreatic buds

Margarita Nieto, Pedro Hevia, Enrique Garcia, Dagmar Klein, Silvia Alvarez-Cubela, Valia Bravo-Egana, Samuel Rosero, Ruth Molano, Nancy Vargas, Camillo Ricordi, Antonello Pileggi, Juan Diez, Juan Dominguez-Bendala, Ricardo Pastori

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

MicroRNAs regulate gene expression by inhibiting translation or inducing target mRNA degradation. MicroRNAs regulate organ differentiation and embryonic development, including pancreatic specification and islet function. We showed previously that miR-7 is highly expressed in human pancreatic fetal and adult endocrine cells. Here we determined the expression profile of miR-7 in the mouse-developing pancreas by RT-PCR and in situ hybridization. MiR-7 expression was low between embryonic days e10.5 and e11.5, then began to increase at e13.5 through e14.5, and eventually decreased by e18. In situ hybridization and immunostaining analysis showed that miR-7 colocalizes with endocrine marker Isl1, suggesting that miR-7 is expressed preferentially in endocrine cells. Whole-mount in situ hybridization shows miR-7 highly expressed in the embryonic neural tube. To investigate the role of miR-7 in development of the mouse endocrine pancreas, antisense miR 7 morpholinos (MO) were delivered to the embryo at an early developmental stage (e10.5 days) via intrauterine fetal heart injection. Inhibition of miR-7 during early embryonic life results in an overall downregulation of insulin production, decreased β-cell numbers, and glucose intolerance in the postnatal period. This phenomenon is specific for miR-7 and possibly due to a systemic effect on pancreatic development. On the other hand, the in vitro inhibition of miR-7 in explanted pancreatic buds leads to β-cell death and generation of β-cells expressing less insulin than those in MO control. Therefore, in addition to the potential indirect effects on pancreatic differentiation derived from its systemic downregulation, the knockdown of miR-7 appears to have a β-cell-specific effect as well. These findings suggest that modulation of miR-7 expression could be utilized in the development of stem cell therapies to cure diabetes.

Original languageEnglish
Pages (from-to)1761-1774
Number of pages14
JournalCell Transplantation
Volume21
Issue number8
DOIs
StatePublished - Nov 5 2012

Fingerprint

Insulin
In Situ Hybridization
Pancreas
Morpholinos
Endocrine Cells
Cells
MicroRNAs
Islets of Langerhans
Down-Regulation
Fetal Heart
Neural Tube
Glucose Intolerance
RNA Stability
Cell death
Medical problems
Cell- and Tissue-Based Therapy
Stem cells
Gene expression
Embryonic Development
Glucose

Keywords

  • β-Cells
  • Antisense microRNA
  • In vivo delivery
  • Islets
  • MicroRNA

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation
  • Biomedical Engineering

Cite this

Antisense miR-7 impairs insulin expression in developing pancreas and in cultured pancreatic buds. / Nieto, Margarita; Hevia, Pedro; Garcia, Enrique; Klein, Dagmar; Alvarez-Cubela, Silvia; Bravo-Egana, Valia; Rosero, Samuel; Molano, Ruth; Vargas, Nancy; Ricordi, Camillo; Pileggi, Antonello; Diez, Juan; Dominguez-Bendala, Juan; Pastori, Ricardo.

In: Cell Transplantation, Vol. 21, No. 8, 05.11.2012, p. 1761-1774.

Research output: Contribution to journalArticle

Nieto, M, Hevia, P, Garcia, E, Klein, D, Alvarez-Cubela, S, Bravo-Egana, V, Rosero, S, Molano, R, Vargas, N, Ricordi, C, Pileggi, A, Diez, J, Dominguez-Bendala, J & Pastori, R 2012, 'Antisense miR-7 impairs insulin expression in developing pancreas and in cultured pancreatic buds', Cell Transplantation, vol. 21, no. 8, pp. 1761-1774. https://doi.org/10.3727/096368911X612521
Nieto M, Hevia P, Garcia E, Klein D, Alvarez-Cubela S, Bravo-Egana V et al. Antisense miR-7 impairs insulin expression in developing pancreas and in cultured pancreatic buds. Cell Transplantation. 2012 Nov 5;21(8):1761-1774. https://doi.org/10.3727/096368911X612521
Nieto, Margarita ; Hevia, Pedro ; Garcia, Enrique ; Klein, Dagmar ; Alvarez-Cubela, Silvia ; Bravo-Egana, Valia ; Rosero, Samuel ; Molano, Ruth ; Vargas, Nancy ; Ricordi, Camillo ; Pileggi, Antonello ; Diez, Juan ; Dominguez-Bendala, Juan ; Pastori, Ricardo. / Antisense miR-7 impairs insulin expression in developing pancreas and in cultured pancreatic buds. In: Cell Transplantation. 2012 ; Vol. 21, No. 8. pp. 1761-1774.
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AU - Bravo-Egana, Valia

AU - Rosero, Samuel

AU - Molano, Ruth

AU - Vargas, Nancy

AU - Ricordi, Camillo

AU - Pileggi, Antonello

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