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LY6H is a specific marker of human pancreatic delta cells

ABSTRACT: LY6H is a specific marker of human pancreatic delta cells

PROVIDER: PRJNA1103695 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28779589_1.fastq.gz	Fastqsanger.gz
	SRR28779589_2.fastq.gz	Fastqsanger.gz
	SRR28779590_1.fastq.gz	Fastqsanger.gz
	SRR28779590_2.fastq.gz	Fastqsanger.gz
	SRR28779591_1.fastq.gz	Fastqsanger.gz

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LY6H is a specific marker of human pancreatic delta cells

Project description:The identification of cell surface markers specific to pancreatic islet cell subsets is important for both the study of islet biology and for investigating the pathophysiology of diseases in which these cell types are lost or damaged. Analysis of publicly available single-cell RNAseq data identified expression of LY6H specifically in the delta cells of the pancreas. This finding was confirmed using immunofluorescence analysis of histological sections of human pancreas, and flow cytometric analysis of human islet preparations. We found that expression of LY6H was restricted to pancreatic delta cells in both control and diabetic donors. Furthermore, we demonstrate that antibodies against LY6H can be used to isolate live delta cell populations from preparations of human islets. This study identified LY6H as a novel cell surface marker of human pancreatic delta cells – a finding that will aid in the identification and characterisation of this important cell type.

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Project description:Human pancreatic δ cells maintain the balance of pancreatic hormones and loss of δ cells or disruption of somatostatin release is associated with diabetes. Despite their important role, human δ cells are scarce (~5% within the islets), limiting physiological studies and drug discovery targeting δ cells. To date, no directed δ-cell differentiation method has been established. In this study, we established a link between fibroblast growth factor (FGF) signaling pathways and pancreatic δ-cell lineage specification. FGF7 helped maintain a pancreatic endoderm/progenitor state with higher expression levels of PDX1 and CHGA, whereas FGF2 biased cells towards the pancreatic δ-cell lineage by inducing the expression of SST and the δ-cell specific transcription factor HHEX, via FGF receptor 1(FGFR1). We developed a differentiation method to generate δ cells from human stem cells by combining FGF2 with FGF7, which synergistically directs pancreatic lineage differentiation and modulates the expression of transcription factors and SST activators during endoderm/endocrine precursor induction. These δ cells display mature RNA profiles and fine secretory granules, secrete somatostatin in response to various stimuli, and suppress insulin secretion from in vitro co-cultured b cells and mouse b cells upon transplantation. The generation of somatostatin-secreting pancreatic δ cells from human stem cells in vitro would provide an unprecedented cell source for drug discovery and cell transplantation studies in diabetes.

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Project description:The identification of cell surface markers specific to pancreatic beta cells is important for both the study of islet biology and for investigating the pathophysiology of diseases in which this cell type is lost or damaged. Following analysis of publicly available single-cell RNAseq data, we identified specific integrin subunits, integrin av and integrin b5, that were expressed in beta cells. Flow cytometry analysis showed that the vast majority of islet cells expressed some level of both the av and b5 subunits. Using our analysis of single-cell sequencing data as a guide, we isolated populations expressing both subunits from isolated human islets and subjected these to bulk RNAseq analysis. This analysis suggested that the avlob5lo population was enriched for endocrine cell types including alpha, delta and gamma cells, whilst the avhib5hi population was enriched for pericytes and stellate cells . Interestingly, the av+b5neg reference population appeared to be enriched for myeloid cells. This finding was further elaborated using immunofluorescence analysis of histological sections derived from donor human pancreas. Despite the broad expression of specific integrin subunits, we found that expression of integrin avb5 heterodimers was restricted to beta cells and that this complex persisted in islet remnants of some type 1 diabetic individuals from which insulin expression had been lost. This study identifies avb5 heterodimers as a novel cell surface marker of human pancreatic beta cells, a finding that will aid in the identification and characterisation of this important cell type.

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