Dysregulated H19/Igf2 expression disrupts cardiac-placental axis during development of Silver-Russell syndrome-like mouse models
- Department of Cell and Developmental Biology, Epigenetics Institute, Perelman School of Medicine, University of Pennsylvania, United States
- Penn Cardiovascular Institute, Perelman School of Medicine, University of Pennsylvania, United States
- Department of Biomedical Sciences, School of Veterinary Medicine, Institute for Regenerative Medicine, University of Pennsylvania, United States
Figures
Figure 1
H19/Igf2 cluster and mouse models utilized in this study.
(A) A schematic representation of the wild-type H19/Igf2 cluster in mouse. The maternally expressed H19 and the paternally expressed Igf2 genes are shown in red and blue, respectively. Black …
Figure 2 with 1 supplement
Growth anomalies and cardiac defects of +/hIC1 embryos.
(A) Fetal weight of the wild-type (blue) and +/hIC1 (red) embryos at E11.5, E12.5, E14.5, E17.5, and E18.5 (mean ± SD). Each data point represents an average weight of each genotype from one litter. …
Figure 2—figure supplement 1
Supplementary data for anomalies observed in +/hIC1 hearts.
(A) Quantification of ventricular wall thickness (µm), measured from E15.5 wild-type and +/hIC1 hearts (mean ± SD). Four wild-type and three +/hIC1 embryos from two different litters were examined. …
Figure 3 with 1 supplement
Placental defects of +/hIC1 embryos.
(A) Placental weight of the wild-type (blue) and +/hIC1 (red) samples at E11.5, E12.5, E14.5, E17.5, and E18.5 (mean ± SD). Each data point represents an average weight of each genotype from one …
Figure 3—figure supplement 1
Supplementary data for anomalies observed in +/hIC1 placentas.
(A) Fetal to placental weight ratios of the wild-type (blue) and +/hIC1 (red) samples at E11.5, E12.5, E14.5, and E15.5. Each data point represents an average ratio of each genotype from one litter. …
Figure 4 with 2 supplements
Normalizing H19 expression through the maternal deletion of H19.
(A) A schematic representation of the rescue breeding between △H19 heterozygous female and hIC1/+ male mice. △H19/hIC1 embryos are expected to express H19 only from the paternal allele and …
Figure 4—figure supplement 1
Characterization of ΔH19 allele.
(A) Targeting strategy to generate the ΔH19 allele. The endogenous H19 locus is shown with restriction sites, gRNA locations used to generate the deletion and binding sites for probes used in …
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Figure 4—figure supplement 1—source data 1
Raw gel images for Figure 4—figure supplement 1B.
- https://cdn.elifesciences.org/articles/78754/elife-78754-fig4-figsupp1-data1-v1.zip
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Figure 4—figure supplement 1—source data 2
Raw gel images for Figure 4—figure supplement 1C.
- https://cdn.elifesciences.org/articles/78754/elife-78754-fig4-figsupp1-data2-v1.zip
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Figure 4—figure supplement 1—source data 3
Raw gel images for Figure 4—figure supplement 1C.
- https://cdn.elifesciences.org/articles/78754/elife-78754-fig4-figsupp1-data3-v1.zip
Figure 4—figure supplement 2
Supplementary data for rescue upon maternal ΔH19 transmission.
(A) Ratio of wild-type, ΔH19/+, +/hIC1, and ΔH19/hIC1 embryos observed in E11.5 and E17.5 litters (>5 pups; mean ± SD). Each data point represents one litter. (B) Left: Relative placental weights of …
Figure 5 with 1 supplement
Restoring H19 and Igf2 expression utilizing maternal H19/Igf2 imprinting control region (ICR) deletion.
(A) A schematic representation of the offspring produced when △3.8 heterozygous female and hIC1/+ male mice are mated is depicted. △3.8/hIC1 embryos are expected to show activation of maternal Igf2 …
Figure 5—figure supplement 1
Supplementary data for rescue upon maternal Δ3.8 transmission.
(A) Left: Relative placental weights of E17.5 wild-type, Δ3.8/+, +/hIC1, and Δ3.8/hIC1 samples, normalized to the average placental weight of the wild-type littermates (mean ± SD). Right: Fetal to …
Figure 6 with 5 supplements
Transcriptomic analysis of E12.5 cardiac endothelial cells from wild-type and mutant embryos.
(A) A gradient H19 and Igf2 expression levels are depicted in E12.5 wild-type, +/hIC1, △H19/+, △H19/hIC1, △3.8/+, and △3.8/hIC1 cardiac endothelial cells. M: male and F: female. (B) Left: Comparison …
Figure 6—figure supplement 1
Supplementary data for differential gene expression of +/hIC1 hearts.
(A) DAPI (4′,6-diamidino-2-phenylindole; blue) on E17.5 wild-type and +/hIC1 hearts from Figure 6C. Images on the right are enlarged from the boxed area of images on the left. Scale bars = 100 µm. (B…
Figure 6—figure supplement 2
Further RNA-seq analysis involving ΔH19/hIC1 endothelial cells.
(A) Volcano plot depicting comparison of ΔH19/hIC1 and wild-type cardiac endothelial cells. (B) Volcano plot depicting comparison of +/hIC1 and ΔH19/hIC1 cardiac endothelial cells. (C) Gene ontology …
Figure 6—figure supplement 3
Enrichment tests for BRIDEAU_IMPRINTED_GENES using gene set enrichment analysis (GSEA).
Normalized enrichment score (NES), nominal p-value, enrichment plot, and expression heatmap of the analyzed gene set comparing (A) +/hIC1 and wild-type, (B) ΔH19/+ and wild-type, (C) ΔH19/hIC1 and …
Figure 6—figure supplement 4
Further RNA-seq analysis utilizing Δ3.8/+ and Δ3.8/hIC1 endothelial cells.
(A) Volcano plot depicting comparison of +/hIC1 and Δ3.8/hIC1 cardiac endothelial cells. (B) Gene ontology (GO) pathways that are enriched for 116 differentially expressed genes (DEGs) that are …
Figure 6—figure supplement 5
Enrichment tests for HALLMARK_HYPOXIA, GROSS_HYPOXIA_VIA_HIF1A_DN using gene set enrichment analysis (GSEA).
Normalized enrichment score (NES), nominal p-value, enrichment plot, and expression heatmap of the (A) HALLMARK_HYPOXIA, (B) GROSS_HYPOXIA_VIA_HIF1A_DN gene set comparing +/hIC1 and wild-type …
Author response image 1
Tetraploid aggregation experiment.
(A) Schematic description of tetraploid aggregation experiment. (B) Level of hIC1 and GFP in embryonic and placental gDNA from hIC1 aggregated samples. (C) Embryonic and placental weight of E17.5 …
Additional files
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Supplementary file 1
Primers and PCR conditions utilized in this study.
- https://cdn.elifesciences.org/articles/78754/elife-78754-supp1-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/78754/elife-78754-mdarchecklist1-v1.docx
