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Series GSE128331 Query DataSets for GSE128331
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
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.
 
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.

 
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
Submission date Mar 14, 2019
Last update date Aug 16, 2022
Contact name Dieter Egli
E-mail(s) de2220@cumc.columbia.edu
Organization name Columbia University
Street address 701 West 168th Street
City New York
State/province New York
ZIP/Postal code 10032
Country USA
 
Platforms (2)
GPL11154 Illumina HiSeq 2000 (Homo sapiens)
GPL18573 Illumina NextSeq 500 (Homo sapiens)
Samples (12)
GSM3671511 Unsorted endocrine cells - single cell RNAseq 1
GSM3671512 Unsorted endocrine cells - single cell RNAseq 2
GSM3671513 Unsorted endocrine cells - single cell RNAseq 3
Relations
BioProject PRJNA527137
SRA SRP188713

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
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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 SelectorHelp
Processed data are available on Series record
Processed data provided as supplementary file
Raw data are available in SRA

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