Transcriptional profiles of human islet and exocrine endothelial cells in subjects with or without impaired glucose metabolism

Jonsson, Ann‐Beth; Hedin, A.; Müller, Malin; Skog, Oskar; Korsgren, Olle

doi:10.1038/s41598-020-79313-y

Cited by 7 publications

(5 citation statements)

References 45 publications

(57 reference statements)

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“…46 Next, we linked overexpressed genes to biological processes (Figure 3c). Acinar tissue was enriched in biological processes, such as translation ribosomal assembly, digestion, and metabolic terms, which is in line with the previous publications, 48,49 and islet samples overexpressed genes related to inter-and intracellular signaling, protein secretion, and hormone-level regulation, as reported previously. 50 In-Depth Proteome Imaging of the Human Islet Microenvironment.…”

Section: ■ Results and Discussionsupporting

confidence: 91%

Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment

Veličković,

Fillmore,

Attah

et al. 2024

Anal. Chem.

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There is increasing interest in developing in-depth proteomic approaches for mapping tissue heterogeneity in a celltype-specific manner to better understand and predict the function of complex biological systems such as human organs. Existing spatially resolved proteomics technologies cannot provide deep proteome coverage due to limited sensitivity and poor sample recovery. Herein, we seamlessly combined laser capture microdissection with a low-volume sample processing technology that includes a microfluidic device named microPOTS (microdroplet processing in one pot for trace samples), multiplexed isobaric labeling, and a nanoflow peptide fractionation approach. The integrated workflow allowed us to maximize proteome coverage of laser-isolated tissue samples containing nanogram levels of proteins. We demonstrated that the deep spatial proteomics platform can quantify more than 5000 unique proteins from a small-sized human pancreatic tissue pixel (∼60,000 μm 2 ) and differentiate unique protein abundance patterns in pancreas. Furthermore, the use of the microPOTS chip eliminated the requirement for advanced microfabrication capabilities and specialized nanoliter liquid handling equipment, making it more accessible to proteomic laboratories.

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Section: ■ Results and Discussionsupporting

confidence: 91%

Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment

Veličković,

Fillmore,

Attah

et al. 2024

Anal. Chem.

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“…There is no transcriptional variation between microvascular endothelial cells sorted from donors with or without impaired glucose metabolism (IGM) [ 35 ]. This suggests that the islet endothelial transcriptome is not per se affected as a consequence of diabetes, i.e.…”

Section: Discussionmentioning

confidence: 99%

Altered microvasculature in pancreatic islets from subjects with type 1 diabetes

Granlund

Hedin²,

Korsgren³

et al. 2022

PLoS ONE

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Aims The transcriptome of different dissociated pancreatic islet cells has been described in enzymatically isolated islets in both health and disease. However, the isolation, culturing, and dissociation procedures likely affect the transcriptome profiles, distorting the biological conclusions. The aim of the current study was to characterize the cells of the islets of Langerhans from subjects with and without type 1 diabetes in a way that reflects the in vivo situation to the highest possible extent. Methods Islets were excised using laser capture microdissection directly from frozen pancreatic tissue sections obtained from organ donors with (n = 7) and without (n = 8) type 1 diabetes. Transcriptome analysis of excised samples was performed using AmpliSeq. Consecutive pancreatic sections were used to estimate the proportion of beta-, alpha-, and delta cells using immunofluorescence and to examine the presence of CD31 positive endothelial regions using immunohistochemistry. Results The proportion of beta cells in islets from subjects with type 1 diabetes was reduced to 0% according to both the histological and transcriptome data, and several alterations in the transcriptome were derived from the loss of beta cells. In total, 473 differentially expressed genes were found in the islets from subjects with type 1 diabetes. Functional enrichment analysis showed that several of the most upregulated gene sets were related to vasculature and angiogenesis, and histologically, vascular density was increased in subjects with type 1 diabetes. Downregulated in type 1 diabetes islets was the gene set epithelial mesenchymal transition. Conclusion A number of transcriptional alterations are present in islets from subjects with type 1 diabetes. In particular, several gene sets related to vasculature and angiogenesis are upregulated and there is an increased vascular density, suggesting an altered microvasculature in islets from subjects with type 1 diabetes. By studying pancreatic islets extracted directly from snap-frozen pancreatic tissue, this study reflects the in vivo situation to a high degree and gives important insights into islet pathophysiology in type 1 diabetes.

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“…To confirm that the capillary 1 EC sub-population corresponded to the islet ECs, we re-analyzed a bulk RNA sequencing dataset ( 57 ) of isolated ECs from endocrine (islets) and exocrine pancreas from 8 adult human donors. Principal component analysis demonstrated that the endocrine sample for donor #8 is an outlier (Fig.…”

Section: Resultsmentioning

confidence: 99%

Uncovering the heterogeneity of pancreatic endothelial cells using integrative and comparative single cell gene expression analysis

Khan

Ahuja

Taïb

et al. 2023

Preprint

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The pancreatic islet vasculature displays tissue-specific physiological and functional adaptations that support rapid glucose sensing and insulin response by beta-cells. To uncover the transcriptomic basis of this specialization, we performed a meta-analysis of multi-organ single cell RNA sequencing atlases employing a unique strategy to avoid transcriptomic contamination. We identified biologically relevant genes involved in sphingosine-1-phosphate-mediated insulin secretion (PLPP1, RDX, CDC42), islet basement membrane formation (SPARC, COL15A1), endothelial cell (EC) permeability (PLVAP, EHD4), membrane transporters (CD320, SLCO2A1) and developmental transcription factors (NKX2-3, AHR). These were validated in silico in an independent dataset. We further established the first integrated transcriptomic atlas of human pancreatic ECs and described two unique capillary subpopulations: exocrine and endocrine pancreas ECs. We validated the spatial localization of key markers using RNAscope and immunofluorescence staining on mouse pancreatic tissue cross-sections. Our findings provide novel insights into pancreatic EC heterogeneity and islet EC function with potential implications in therapeutic strategies.

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Transcriptional profiles of human islet and exocrine endothelial cells in subjects with or without impaired glucose metabolism

Cited by 7 publications

References 45 publications

Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment

Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment

Altered microvasculature in pancreatic islets from subjects with type 1 diabetes

Uncovering the heterogeneity of pancreatic endothelial cells using integrative and comparative single cell gene expression analysis

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