Complex aneuploidy triggers autophagy and p53-mediated apoptosis and impairs the second lineage segregation in human preimplantation embryos
- Brussels Health Campus/Faculty of Medicine and Pharmacy, Research Group Genetics Reproduction and Development, Vrije Universiteit Brussel, Belgium
- Brussels Health Campus, Brussels IVF, Universitair Ziekenhuis Brussel (UZ Brussel), Belgium
- Brussels Health Campus, Medical Genetics, Universitair Ziekenhuis Brussel (UZ Brussel), Belgium
Figures
Figure 1 with 1 supplement
RNA-sequencing of trophectoderm cells reveals gene dosage defects and transcriptomic signatures of p53 activation and apoptosis.
(a) Principal component analysis after transcriptome analysis of aneuploid versus euploid embryos. T=Trisomy, M=Monosomy. (b) Comparison between the diagnosis of aneuploid embryos obtained after …
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Figure 1—source data 1
Count table of RNA-sequencing data.
- https://cdn.elifesciences.org/articles/88916/elife-88916-fig1-data1-v1.zip
Figure 1—figure supplement 1
Expanded RNA-sequencing analysis.
(a) Principal component analysis of reversine versus euploid embryos. (b) InferCNV analysis of individual euploid samples compared to the reference set. (c) InferCNV analysis of individual reversine …
Figure 2
Human embryos with the highest number of genes with abnormal copy number show stronger p53 pathway and apoptosis response.
(a) Principal component analysis after transcriptome analysis of high-dosage aneuploid versus euploid embryos. T=Trisomy, M=Monosomy. (b) Volcano plot after differential gene expression analysis …
Figure 3 with 1 supplement
Immunostaining reveals increased proteotoxic stress, DNA-damage independent p53-activation, autophagy, and apoptosis.
(a) Orthogonal projections after immunostaining of euploid and aneuploid embryos for DNA (white) and CASP3/7 (green). T=Trisomy, M=Monosomy. Source: Experiment 1. (b) Orthogonal projections after …
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Figure 3—source data 1
Raw data of figures.
- https://cdn.elifesciences.org/articles/88916/elife-88916-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
S15-p53 expression in trophectoderm (TE) vs OCT4-positive cells (inner cell mass, ICM/epiblast, EPI) in aneuploid embryos.
(a) Orthogonal projections after immunostaining of euploid and aneuploid embryos for DNA (white), S15p53 (turquoise), and OCT4 (magenta). (b) Percentages of nuclei positive for S15p53 per embryo in …
Figure 4
Aneuploid human embryos show less cells in trophectoderm and OCT4-positive cells and impaired lineage segregation events.
(a) Orthogonal projections after immunostaining of euploid (n=7) and aneuploid embryos (n=10) for DNA (white) and NANOG (green). T=Trisomy. Source: Experiment 3. (b) NANOG nuclear enrichment in …
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Figure 4—source data 1
Raw data of figures.
- https://cdn.elifesciences.org/articles/88916/elife-88916-fig4-data1-v1.xlsx
Author response image 1
Author response image 2
Author response image 3
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-HSP70 (Mouse monoclonal) | Invitrogen | MA3-007 | IF(1:200) |
| Antibody | Anti-SQSTM1/p62 (Mouse monoclonal) | Abcam | ab56416 | IF(1:100) |
| Antibody | Recombinant Anti-LC3B (Rabbit monoclonal) | Abcam | ab192890 | IF(1:838) |
| Antibody | Anti-OCT3/4 (Mouse monoclonal) | Santa-Cruz | sc-5279 | IF(1:200) |
| Antibody | Anti-human phosphor-p53 S15 (Rabbit polyclonal) | R&D | AF1043 | IF(1:100) |
| Antibody | Anti-GATA4 (Rat monoclonal) | Invitrogen | 14-9980-82 | IF(1:400) |
| Antibody | Anti-DRAM (Rabbit polyclonal) | Invitrogen | PA20-335 | IF(1:500) |
| Antibody | Anti-phospho-histone H2A.X Ser139 (Mouse monoclonal) | Sigma-Aldrich | 05–636 | IF(1:500) |
| Antibody | Anti-NANOG (Goat polyclonal) | R&D | AF1197 | IF(1:200) |
| Chemical compound, drug | Reversine | Stem Cell Technologies | # 72614 | 0.5 µM |
| Other | CaspGLOW Fluorescein Active Caspase-8 Staining Kit | Invitrogen | 88-7005-42 | IF(1:300) |
| Other | CellEvent Caspase-3/7 Green Detection Reagent | Invitrogen | C10423 | IF(1:400) |
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/88916/elife-88916-mdarchecklist1-v1.docx
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Supplementary file 1
PGT diagnoses and inferCNV results first experiment.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp1-v1.xlsx
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Supplementary file 2
Differentially expressed genes aneuploid versus euploid embryos.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp2-v1.xlsx
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Supplementary file 3
Differentially expressed genes reversine versus euploid embryos.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp3-v1.xlsx
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Supplementary file 4
Ranks of imbalanced gene loci for the first experiment.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp4-v1.xlsx
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Supplementary file 5
Differentially expressed genes high-dosage versus euploid embryos.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp5-v1.xlsx
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Supplementary file 6
Gene dosage ranking Experiment 3 and 4.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp6-v1.xlsx
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Supplementary file 7
PGT diagnoses experiment 4.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp7-v1.xlsx
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Supplementary file 8
Gene dosage classification experiment 4.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp8-v1.xlsx
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Supplementary file 9
Antibodies and reagents used in this study.
- https://cdn.elifesciences.org/articles/88916/elife-88916-supp9-v1.xlsx
