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| Status |
Public on Jun 19, 2019 |
| Title |
Point mutations in the PDX1 transactivation domain impair human β-cell development and function (ChIP-Seq) |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
Missense mutations in coding region of PDX1 predispose to type-2 diabetes mellitus as well as cause MODY through largely unexplored mechanisms. Here, we screened a large cohort of subjects with increased risk for diabetes and identified two subjects with impaired glucose tolerance carrying heterozygous missense mutations in the PDX1 coding region leading to single amino acid exchanges (P33T, C18R) in its transactivation domain. We generated iPSCs from patients with heterozygous PDX1P33T/+, PDX1C18R/+ mutations and engineered isogenic cell lines carrying homozygous PDX1P33T/P33T, PDX1C18R/C18R mutations and a heterozygous PDX1 loss-of-function mutation (PDX1+/-). Using an in vitro β-cell differentiation protocol, we demonstrated that both PDX1P33T/+, PDX1C18R/+ and PDX1P33T/P33T, PDX1C18R/C18R mutations impair β-cell differentiation and function. Furthermore, PDX1+/- and PDX1P33T/P33T mutations reduced differentiation efficiency of pancreatic progenitors (PPs), due to downregulation of PDX1-bound genes, including transcription factors MNX1 and PDX1 as well as insulin resistance gene CES1. Additionally, both PDX1P33T/+ and PDX1P33T/P33T mutations in PPs reduced the expression of PDX1-bound genes including the long-noncoding RNA, MEG3 and the imprinted gene NEURONATIN, both involved in insulin synthesis and secretion. Our results reveal mechanistic details of how diabetes-associated PDX1 point mutations impair human pancreatic endocrine lineage formation and β-cell function and contribute to pre-disposition for diabetes.
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| Overall design |
We performed ChIP-seq of PDX1 and H3K27ac on PDX1P33T/+ cells at PP stage
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| Contributor(s) |
Wang X, Sterr M, Ansarullah A, Burtscher I, Böttcher A, Beckenbauer J, Siehler J, Staiger H, Häring H, Lederer G, Meitinger T, Cernilogar F, Schotta G, Irmler M, Beckers J, Wright C, Bakhti M, Lickert H |
| Citation(s) |
30930126 |
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| Submission date |
Jan 28, 2019 |
| Last update date |
Jun 19, 2019 |
| Contact name |
Heiko Lickert |
| E-mail(s) |
heiko.lickert@helmholtz-munich.de
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| Organization name |
Helmholtz Zentrum München German Research Center for Environmental Health
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| Department |
Institute of Diabetes and Regeneration Research
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| Street address |
Ingolstaedter Landstraße 1
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| City |
Neuherberg |
| ZIP/Postal code |
85764 |
| Country |
Germany |
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| Platforms (1) |
| GPL18460 |
Illumina HiSeq 1500 (Homo sapiens) |
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| Samples (5)
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| This SubSeries is part of SuperSeries: |
| GSE125770 |
Point mutations in the PDX1 transactivation domain impair human β-cell development and function |
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| Relations |
| BioProject |
PRJNA517474 |
| SRA |
SRP182693 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE125768_P33T_PDX1_Multi-Condidtion_GEM_events.bed.gz |
130.5 Kb |
(ftp)(http) |
BED |
| GSE125768_XM001-P33T_H3K27ac_HOMER_Peaks.bed.gz |
530.1 Kb |
(ftp)(http) |
BED |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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