Abstract
BackgroundHuman embryonic stem cells represent a potentially unlimited source of insulin-producing cells for diabetes therapy. While tremendous progress has been made in directed differentiation of human embryonic stem cells into IPCs in vitro, the mechanisms controlling its differentiation and function are not fully understood. Previous studies revealed that lysine-specific demethylase 1(LSD1) balanced the self-renewal and differentiation in human induced pluripotent stem cells and human embryonic stem cells. This study aims to explore the role of LSD1 in directed differentiation of human embryonic stem cells into insulin-producing cells.MethodsHuman embryonic stem cell line H9 was induced into insulin-producing cells by a four-step differentiation protocol. Lentivirus transfection was applied to knockdown LSD1 expression. Immunofluorescence assay and flow cytometry were utilized to check differentiation efficiency. Western blot was used to examine signaling pathway proteins and differentiation-associated proteins. Insulin/C-peptide release was assayed by ELISA. Statistical analysis between groups was carried out with one-way ANOVA tests or a student’s t test when appropriate.ResultsInhibition or silencing LSD1 promotes the specification of pancreatic progenitors and finally the commitment of functional insulin-producing β cells; Moreover, inhibition or silencing LSD1 activated ERK signaling and upregulated pancreatic progenitor associated genes, accelerating pre-maturation of pancreatic progenitors, and conferred the NKX6.1+ population with better proliferation ability. IPCs with LSD1 inhibitor tranylcypromine treatment displayed enhanced insulin secretion in response to glucose stimulation.ConclusionsWe identify a novel role of LSD1 inhibition in promoting IPCs differentiation from hESCs, which would be emerged as potential intervention for generation of functional pancreatic β cells to cure diabetes.
Highlights
Diabetes mellitus is a chronic metabolic syndrome featured by elevated blood glucose levels results from inadequate production of the hormone insulin
Given that Lysine-specific demethylase 1 (LSD1) expression was gradually reduced during pancreatic development, we carried out the following inactivation experiments to investigate whether LSD1 played a role in the Insulin-producing cells (IPCs) differentiation of Human embryonic stem cells (hESCs) and in which stage LSD1 was most effective
In summary, our study demonstrates that knockdown or inhibition of LSD1 promoted pancreatic progenitors differentiation of hESCs through activating ERK signaling, and sustained TCP treatment improved the IPCs maturation witnessed by better glucose-stimulated insulin secretion (GSIS)
Summary
Diabetes mellitus is a chronic metabolic syndrome featured by elevated blood glucose levels results from inadequate production of the hormone insulin. It was estimated over 451 million adults have diabetes, and diabetes has been a major health problem worldwide [1]. Human embryonic stem cells (hESCs) represent an attractive source for β-cells production, owing to its pluripotency and unlimited proliferation features [4]. Human embryonic stem cells represent a potentially unlimited source of insulin-producing cells for diabetes therapy. This study aims to explore the role of LSD1 in directed differentiation of human embryonic stem cells into insulin-producing cells
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