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Biology subjects

de Koning, E. J.

Publications and source records attributed to de Koning, E. J..

3 recordsLinked to original sources

Cell type-specific eQTL detection from single-cell RNA-seq reveals post-transcriptional regulatory mechanisms in human islets

Gene regulatory networks (GRNs) must be robust to maintain cellular identity across individuals yet flexible to accommodate population genetic variants. Comparing expression quantitative trait loci (eQTLs) in health and disease can therefore reveal hidden players in gene regulation. Here, we developed a computational pipeline to infer post-transcriptional regulatory events and their functional consequences from eQTL signals located in the 3' untranslated region of genes using single-cell RNA sequencing. As a case study, we repurposed datasets from human islets of donors with and without type 2 diabetes (T2D). We identified eQTL landscapes differing by cell type and diabetes status, with cell-type specificity associated with gene differential expression and RNA-binding proteins. Integrating eQTLs with GWAS and miRNA hits linked variants in G6PC2, QDPR, and RELL1 to insulin secretion, endoplasmic reticulum stress, and oxidative phosphorylation. Our pipeline provides a framework to unravel post-transcriptional regulatory mechanisms in health and disease at cell type resolution.

genetics↗

Oxidative stress in alpha and beta cells as a selection criterion for biocompatible biomaterials

The clinical success of islet transplantation is limited by factors including acute ischemia, stress upon transplantation, and delayed vascularization. Islets experience high levels of oxidative stress due to delayed vascularization after transplantation and this can be further aggravated by their encapsulation and undesirable cell-biomaterial interactions. To identify biomaterials that would not further increase oxidative stress levels and that are also suitable for manufacturing a beta cell encapsulation device, we studied five clinically approved polymers for their effect on oxidative stress and islet (alpha and beta cell) function. We found that 300 poly(ethylene oxide terephthalate) 55/poly(butylene terephthalate) 45 (PEOT/PBT300) was more resistant to breakage and more elastic than other biomaterials, which is important for its immunoprotective function. In addition, PEOT/PBT300 did not induce oxidative stress or reduce viability in MIN6 beta cells, and even promoted protective endogenous antioxidant expression over 7 days. Importantly, PEOT/PBT300 is one of the biomaterials we studied that did not interfere with insulin secretion in human islets. These data indicate that PEOT/PBT300 may be a suitable biomaterial for an islet encapsulation device.

cell biology↗

Cell type purification by single-cell transcriptome-trained sorting

Traditional cell type enrichment using fluorescence activated cell sorting (FACS) relies on methods that specifically label the cell type of interest. Here we propose GateID, a computational method that combines single-cell transcriptomics for unbiased cell type identification with FACS index sorting to purify cell types of choice. We validate GateID by purifying various cell types from the zebrafish kidney marrow and the human pancreas without resorting to specific antibodies or transgenes.

bioinformatics↗