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| Status |
Public on Mar 16, 2019 |
| Title |
HNF1A deficiency impairs β-cell fate, granule maturation and function |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Mutations in HNF1A cause Maturity Onset Diabetes of the Young type 3, the second most frequent form of diabetes caused by single gene mutation. We generated human pancreatic stem cell-derived endocrine cells with mutations in HNF1A and show that HNF1A deficiency impairs scβ-cell fate, insulin granule maturation and the secretion of insulin in a glucose responsive manner. Single-cell RNA sequencing reveals that HNF1A orchestrates a network of genes involved in glucose metabolism, zinc transport, calcium ion binding and hormone exocytosis. Furthermore, in both patients and stem cell-derived β-cells, HNF1A deficiency altered the stoichiometry of secreted c-peptide to insulin. Sulfonylurea, used in the treatment of these patients, restored both insulin secretion and stoichiometry. Significantly, uncoupling of c-peptide and insulin secretion as described here questions the common practice in using c-peptide as a proxy to evaluate β-cell function. We also demonstrate that correction of the HNF1A locus restores function, providing a path to cell therapy.
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| Overall design |
Human embryonic stem cells with different HNF1A genotypes (WT, KO1, KO2, R200Q homozygous) were differentiated into islet-like clusters of endocrine cells for 27-28 days in vitro. First set of 6 samples, clusters of islet-like cells were dissociated into single cells and analyzed by single cell RNA sequencing. There are two WT, two KO and two R200Q samples.
Second set of 5 samples, clusters of islet-like cells were dissociated into single cells and insulin producing cells were purified by FACS sorting for INS-GFP. Cells were analyzed by bulk RNA sequencing. There are three WT and two KO.
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| Contributor(s) |
González BJ, Zhao H, Lee J, LeDuc CA, Goulbourne CN, Chen X, Chung WK, Jurczyk A, Gromada J, Shen Y, Goland RS, Leibel RL, Egli D |
| Citation(s) |
35918471 |
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| Submission date |
Mar 14, 2019 |
| Last update date |
Aug 16, 2022 |
| Contact name |
Dieter Egli |
| E-mail(s) |
de2220@cumc.columbia.edu
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| Organization name |
Columbia University
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| Street address |
701 West 168th Street
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| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10032 |
| Country |
USA |
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| Platforms (2) |
| GPL11154 |
Illumina HiSeq 2000 (Homo sapiens) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (12)
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| GSM3671511 |
Unsorted endocrine cells - single cell RNAseq 1 |
| GSM3671512 |
Unsorted endocrine cells - single cell RNAseq 2 |
| GSM3671513 |
Unsorted endocrine cells - single cell RNAseq 3 |
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| Relations |
| BioProject |
PRJNA527137 |
| SRA |
SRP188713 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE128331_Cell_labels_sc_cleaned.csv.gz |
106.0 Kb |
(ftp)(http) |
CSV |
| GSE128331_RAW.tar |
5.5 Mb |
(http)(custom) |
TAR (of CSV) |
| GSE128331_Raw_counts_sc_cleaned.csv.gz |
78.1 Mb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Processed data are available on Series record |
| Processed data provided as supplementary file |
| Raw data are available in SRA |
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