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Modelling and rescuing neurodevelopmental defect of Down syndrome using induced pluripotent stem cells from monozygotic twins discordant for trisomy 21.

Hibaoui Y, Grad I, Letourneau A, Sailani MR, Dahoun S, Santoni FA, Gimelli S, Guipponi M, Pelte MF, Béna F, Antonarakis SE, Feki A - EMBO Mol Med (2013)

Bottom Line: Here, we report the generation and characterization of induced pluripotent stem cells (iPSCs) derived from monozygotic twins discordant for trisomy 21 in order to eliminate the effects of the variability of genomic background.These defects were associated with changes in the architecture and density of neurons, astroglial and oligodendroglial cells together with misexpression of genes involved in neurogenesis, lineage specification and differentiation.Importantly, we found that targeting DYRK1A pharmacologically or by shRNA results in a considerable correction of these defects.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Research Laboratory Department of Obstetrics and Gynecology, Geneva University Hospitals, Geneva, Switzerland.

ABSTRACT
Down syndrome (trisomy 21) is the most common viable chromosomal disorder with intellectual impairment and several other developmental abnormalities. Here, we report the generation and characterization of induced pluripotent stem cells (iPSCs) derived from monozygotic twins discordant for trisomy 21 in order to eliminate the effects of the variability of genomic background. The alterations observed by genetic analysis at the iPSC level and at first approximation in early development illustrate the developmental disease transcriptional signature of Down syndrome. Moreover, we observed an abnormal neural differentiation of Down syndrome iPSCs in vivo when formed teratoma in NOD-SCID mice, and in vitro when differentiated into neuroprogenitors and neurons. These defects were associated with changes in the architecture and density of neurons, astroglial and oligodendroglial cells together with misexpression of genes involved in neurogenesis, lineage specification and differentiation. Furthermore, we provide novel evidence that dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1A (DYRK1A) on chromosome 21 likely contributes to these defects. Importantly, we found that targeting DYRK1A pharmacologically or by shRNA results in a considerable correction of these defects.

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Hematoxylin and eosin staining analysis of teratoma generated after intramuscular injection of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) into SCID mice. See also supplementary Fig S4.Spontaneous in vitro differentiation of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) as embryoid bodies (EBs) in suspension culture for 4 days and as adherent cells for an additional 17 days. These EBs expressed α-SMA (mesoderm), AFP (endoderm) and β3-tubulin (ectoderm).
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fig03: Hematoxylin and eosin staining analysis of teratoma generated after intramuscular injection of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) into SCID mice. See also supplementary Fig S4.Spontaneous in vitro differentiation of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) as embryoid bodies (EBs) in suspension culture for 4 days and as adherent cells for an additional 17 days. These EBs expressed α-SMA (mesoderm), AFP (endoderm) and β3-tubulin (ectoderm).

Mentions: To document their developmental potential in vivo, iPSCs were injected intramuscularly into immunodeficient SCID mice. Histological analysis revealed that Twin-N-iPSCs formed teratoma with all embryonic germ layers: mesoderm, ectoderm and endoderm (Fig 3A upper panel and supplementary Fig S4). In contrast, injection of Twin-DS-iPSCs formed teratoma with multiple cysts which were surrounded by an abundant undifferentiated mesenchyme (Fig 3A lower panel). Mucinous intestinal-type differentiation or foci of bone were also found (Fig 3A lower panel). Surprisingly, differentiated ectoderm germ structures were not found in Twin-DS-iPSC-derived teratomas (Fig 3A lower panel). These results were confirmed by the near absence of staining for the neuroepithelial marker NESTIN in Twin-DS-iPSC-derived teratoma (supplementary Fig S4). We next determined that these iPSCs could differentiate in vitro into all three embryonic germ layers as detected by expression of the ectodermal marker β3-TUBULIN, the mesodermal marker α-SMOOTH MUSCLE ACTIN (α-SMA) and the endodermal marker α-FETOPROTEIN (AFP; Fig 3B).


Modelling and rescuing neurodevelopmental defect of Down syndrome using induced pluripotent stem cells from monozygotic twins discordant for trisomy 21.

Hibaoui Y, Grad I, Letourneau A, Sailani MR, Dahoun S, Santoni FA, Gimelli S, Guipponi M, Pelte MF, Béna F, Antonarakis SE, Feki A - EMBO Mol Med (2013)

Hematoxylin and eosin staining analysis of teratoma generated after intramuscular injection of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) into SCID mice. See also supplementary Fig S4.Spontaneous in vitro differentiation of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) as embryoid bodies (EBs) in suspension culture for 4 days and as adherent cells for an additional 17 days. These EBs expressed α-SMA (mesoderm), AFP (endoderm) and β3-tubulin (ectoderm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927959&req=5

fig03: Hematoxylin and eosin staining analysis of teratoma generated after intramuscular injection of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) into SCID mice. See also supplementary Fig S4.Spontaneous in vitro differentiation of Twin-N-iPSCs (upper panel) and Twin-DS-iPSCs (lower panel) as embryoid bodies (EBs) in suspension culture for 4 days and as adherent cells for an additional 17 days. These EBs expressed α-SMA (mesoderm), AFP (endoderm) and β3-tubulin (ectoderm).
Mentions: To document their developmental potential in vivo, iPSCs were injected intramuscularly into immunodeficient SCID mice. Histological analysis revealed that Twin-N-iPSCs formed teratoma with all embryonic germ layers: mesoderm, ectoderm and endoderm (Fig 3A upper panel and supplementary Fig S4). In contrast, injection of Twin-DS-iPSCs formed teratoma with multiple cysts which were surrounded by an abundant undifferentiated mesenchyme (Fig 3A lower panel). Mucinous intestinal-type differentiation or foci of bone were also found (Fig 3A lower panel). Surprisingly, differentiated ectoderm germ structures were not found in Twin-DS-iPSC-derived teratomas (Fig 3A lower panel). These results were confirmed by the near absence of staining for the neuroepithelial marker NESTIN in Twin-DS-iPSC-derived teratoma (supplementary Fig S4). We next determined that these iPSCs could differentiate in vitro into all three embryonic germ layers as detected by expression of the ectodermal marker β3-TUBULIN, the mesodermal marker α-SMOOTH MUSCLE ACTIN (α-SMA) and the endodermal marker α-FETOPROTEIN (AFP; Fig 3B).

Bottom Line: Here, we report the generation and characterization of induced pluripotent stem cells (iPSCs) derived from monozygotic twins discordant for trisomy 21 in order to eliminate the effects of the variability of genomic background.These defects were associated with changes in the architecture and density of neurons, astroglial and oligodendroglial cells together with misexpression of genes involved in neurogenesis, lineage specification and differentiation.Importantly, we found that targeting DYRK1A pharmacologically or by shRNA results in a considerable correction of these defects.

View Article: PubMed Central - PubMed

Affiliation: Stem Cell Research Laboratory Department of Obstetrics and Gynecology, Geneva University Hospitals, Geneva, Switzerland.

ABSTRACT
Down syndrome (trisomy 21) is the most common viable chromosomal disorder with intellectual impairment and several other developmental abnormalities. Here, we report the generation and characterization of induced pluripotent stem cells (iPSCs) derived from monozygotic twins discordant for trisomy 21 in order to eliminate the effects of the variability of genomic background. The alterations observed by genetic analysis at the iPSC level and at first approximation in early development illustrate the developmental disease transcriptional signature of Down syndrome. Moreover, we observed an abnormal neural differentiation of Down syndrome iPSCs in vivo when formed teratoma in NOD-SCID mice, and in vitro when differentiated into neuroprogenitors and neurons. These defects were associated with changes in the architecture and density of neurons, astroglial and oligodendroglial cells together with misexpression of genes involved in neurogenesis, lineage specification and differentiation. Furthermore, we provide novel evidence that dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1A (DYRK1A) on chromosome 21 likely contributes to these defects. Importantly, we found that targeting DYRK1A pharmacologically or by shRNA results in a considerable correction of these defects.

Show MeSH
Related in: MedlinePlus