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Diabetologia
Published in: Diabetologia 12/2016

01-12-2016 | Article

Sequential cleavage of insulin receptor by calpain 2 and γ-secretase impairs insulin signalling

Authors: Tomoyuki Yuasa, Kikuko Amo-Shiinoki, Shuhei Ishikura, Mitsuyoshi Takahara, Takaaki Matsuoka, Hideaki Kaneto, Akio Kuroda, Munehide Matsuhisa, Seiichi Hashida

Published in: Diabetologia | Issue 12/2016

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Abstract

Aims/hypothesis

Soluble insulin receptor (sIR), the ectodomain of the insulin receptor (IR), has been detected in human plasma and its concentration paralleled that of blood glucose. We have previously developed an in vitro model using HepG2 liver-derived cells, which mimics changes in sIR levels in plasma from diabetic patients and shows that calcium-dependent proteases cleave IR extracellularly (a process known as shedding). The present study aimed to reveal the mechanisms of IR cleavage.

Methods

Using the in vitro model, we investigated the molecular mechanisms of IR cleavage, which is accelerated by high-glucose treatment. We also analysed the relationship between IR cleavage and cellular insulin resistance, and the correlation between plasma sIR levels and insulin sensitivity, which was assessed by the euglycaemic–hyperinsulinaemic clamp technique.

Results

Here, we determined that calpain 2, which is secreted into the extracellular space associated with exosomes, directly cleaved the ectodomain of the IRβ subunit (IRβ), which in turn promoted intramembrane cleavage of IRβ by γ-secretase. IR cleavage impaired insulin signalling and the inhibition of IR cleavage (by knockdown of calpain 2 and γ-secretase), restored IR substrate-1 and Akt, independent of IR. Furthermore, the glucose-lowering drug, metformin, prevented IR cleavage accompanied by inhibition of calpain 2 release in exosomes, and re-established insulin signalling. In patients with type 2 diabetes, plasma sIR levels inversely correlated with insulin sensitivity.

Conclusions/interpretation

Sequential cleavage of IR by calpain 2 and γ-secretase may contribute to insulin signalling in cells and its inhibition may be partly responsible for the glucose-lowering effects of metformin. Thus, IR cleavage may offer a new mechanism for the aetiology of insulin resistance.
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Metadata
Title
Sequential cleavage of insulin receptor by calpain 2 and γ-secretase impairs insulin signalling
Authors
Tomoyuki Yuasa
Kikuko Amo-Shiinoki
Shuhei Ishikura
Mitsuyoshi Takahara
Takaaki Matsuoka
Hideaki Kaneto
Akio Kuroda
Munehide Matsuhisa
Seiichi Hashida
Publication date
01-12-2016
Publisher
Springer Berlin Heidelberg
Published in
Diabetologia / Issue 12/2016
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-016-4102-5

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