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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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Karyotypes of Twin-N-iPSCs and Twin-DS-iPSCs are 46, XX and 47, XX+21, respectively.Ideogram and array-CGH profile showing the gain of one DNA copy of chromosome 21 in Twin-DS-iPSCs in comparison with Twin-N-iPSCs.Comparison of the normalized gene expression levels mean of log2 RPKM between Twin-N-iPSCs and Twin-DS-iPSCs (HSA21 genes are shown in red and the rest of the genome in blue).Principal component analysis (PCA) plot based on the normalized expression values in Twin-N-iPSCs and Twin-DS-iPSCs. The percentages represent the proportion of variance explained by each component.Heat map of the normalized gene expression values in Twin-DS-iPSCs and Twin N-iPSCs for the 1204 differentially expressed genes. A negative z-score (in red) indicates low expression (below the mean) whereas a positive z-score (in green) shows high expression (above the mean). Diagram showing the proportion of genes differentially expressed between Twin-N-iPSCs and Twin-DS-iPSCs. See also supplementary Table S2.
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fig02: Karyotypes of Twin-N-iPSCs and Twin-DS-iPSCs are 46, XX and 47, XX+21, respectively.Ideogram and array-CGH profile showing the gain of one DNA copy of chromosome 21 in Twin-DS-iPSCs in comparison with Twin-N-iPSCs.Comparison of the normalized gene expression levels mean of log2 RPKM between Twin-N-iPSCs and Twin-DS-iPSCs (HSA21 genes are shown in red and the rest of the genome in blue).Principal component analysis (PCA) plot based on the normalized expression values in Twin-N-iPSCs and Twin-DS-iPSCs. The percentages represent the proportion of variance explained by each component.Heat map of the normalized gene expression values in Twin-DS-iPSCs and Twin N-iPSCs for the 1204 differentially expressed genes. A negative z-score (in red) indicates low expression (below the mean) whereas a positive z-score (in green) shows high expression (above the mean). Diagram showing the proportion of genes differentially expressed between Twin-N-iPSCs and Twin-DS-iPSCs. See also supplementary Table S2.

Mentions: These iPSCs were evaluated to confirm the disease-specific genotype of their parental somatic cells. As revealed by karyotype and array-based comparative genomic hybridization analysis, Twin-DS-iPSCs showed the characteristic trisomy 21 while Twin-N-iPSCs had a normal karyotype (Fig 2A and B). Then, whole transcriptome analysis using mRNA-Sequencing confirmed that the majority of HSA21 genes are indeed more expressed in the trisomic lines than the euploid lines (Fig 2C and supplementary Fig S2A) which is consistent with the overall up-regulation of HSA21 genes in individuals with DS (Antonarakis et al, 2004; Lott & Dierssen, 2010). Principal component analysis (PCA) showed that the first principal component (PC1) clearly discriminated the trisomic from the euploid samples, explaining most of the variance in the whole genome data (72%) and in the analysis of HSA21 genes (91%; Fig 2D and supplementary Fig S2B).


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)

Karyotypes of Twin-N-iPSCs and Twin-DS-iPSCs are 46, XX and 47, XX+21, respectively.Ideogram and array-CGH profile showing the gain of one DNA copy of chromosome 21 in Twin-DS-iPSCs in comparison with Twin-N-iPSCs.Comparison of the normalized gene expression levels mean of log2 RPKM between Twin-N-iPSCs and Twin-DS-iPSCs (HSA21 genes are shown in red and the rest of the genome in blue).Principal component analysis (PCA) plot based on the normalized expression values in Twin-N-iPSCs and Twin-DS-iPSCs. The percentages represent the proportion of variance explained by each component.Heat map of the normalized gene expression values in Twin-DS-iPSCs and Twin N-iPSCs for the 1204 differentially expressed genes. A negative z-score (in red) indicates low expression (below the mean) whereas a positive z-score (in green) shows high expression (above the mean). Diagram showing the proportion of genes differentially expressed between Twin-N-iPSCs and Twin-DS-iPSCs. See also supplementary Table S2.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig02: Karyotypes of Twin-N-iPSCs and Twin-DS-iPSCs are 46, XX and 47, XX+21, respectively.Ideogram and array-CGH profile showing the gain of one DNA copy of chromosome 21 in Twin-DS-iPSCs in comparison with Twin-N-iPSCs.Comparison of the normalized gene expression levels mean of log2 RPKM between Twin-N-iPSCs and Twin-DS-iPSCs (HSA21 genes are shown in red and the rest of the genome in blue).Principal component analysis (PCA) plot based on the normalized expression values in Twin-N-iPSCs and Twin-DS-iPSCs. The percentages represent the proportion of variance explained by each component.Heat map of the normalized gene expression values in Twin-DS-iPSCs and Twin N-iPSCs for the 1204 differentially expressed genes. A negative z-score (in red) indicates low expression (below the mean) whereas a positive z-score (in green) shows high expression (above the mean). Diagram showing the proportion of genes differentially expressed between Twin-N-iPSCs and Twin-DS-iPSCs. See also supplementary Table S2.
Mentions: These iPSCs were evaluated to confirm the disease-specific genotype of their parental somatic cells. As revealed by karyotype and array-based comparative genomic hybridization analysis, Twin-DS-iPSCs showed the characteristic trisomy 21 while Twin-N-iPSCs had a normal karyotype (Fig 2A and B). Then, whole transcriptome analysis using mRNA-Sequencing confirmed that the majority of HSA21 genes are indeed more expressed in the trisomic lines than the euploid lines (Fig 2C and supplementary Fig S2A) which is consistent with the overall up-regulation of HSA21 genes in individuals with DS (Antonarakis et al, 2004; Lott & Dierssen, 2010). Principal component analysis (PCA) showed that the first principal component (PC1) clearly discriminated the trisomic from the euploid samples, explaining most of the variance in the whole genome data (72%) and in the analysis of HSA21 genes (91%; Fig 2D and supplementary Fig S2B).

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