Small molecule modulation of insulin receptor-insulin like growth factor-1 receptor heterodimers in human endothelial cells

IF 3.8 3区医学 Q2 CELL BIOLOGY

Molecular and Cellular Endocrinology Pub Date : 2024-10-16 DOI:10.1016/j.mce.2024.112387

Chloe G. Myers , Hema Viswambharan , Natalie J. Haywood , Katherine Bridge , Samuel Turvey , Tom Armstrong , Lydia Lunn , Paul J. Meakin , Karen E. Porter , Eva M. Clavane , David J. Beech , Richard M. Cubbon , Stephen B. Wheatcroft , Martin J. McPhillie , Tarik Issad , Colin WG. Fishwick , Mark T. Kearney , Katie J. Simmons

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引用次数: 0

Abstract

Objectives

The insulin receptor (IR) and insulin like growth factor-1 receptor (IGF-1R) are heterodimers consisting of two extracellular α-subunits and two transmembrane β -subunits. Insulin αβ and insulin like growth factor-1 αβ hemi-receptors can heterodimerize to form hybrids composed of one IR αβ and one IGF-1R αβ. The function of hybrids in the endothelium is unclear. We sought insight by developing a small molecule capable of reducing hybrid formation in endothelial cells.

Methods

We performed a high-throughput small molecule screening, based on a homology model of the apo hybrid structure. Endothelial cells were studied using western blotting and qPCR to determine the effects of small molecules that reduced hybrid formation.

Results

Our studies unveil a first-in-class quinoline-containing heterocyclic small molecule that reduces hybrids by >50% in human umbilical vein endothelial cells (HUVECs) with no effects on IR or IGF-1R. This small molecule reduced expression of the negative regulatory p85α subunit of phosphatidylinositol 3-kinase, increased basal phosphorylation of the downstream target Akt and enhanced insulin/insulin-like growth factor-1 and shear stress-induced serine phosphorylation of Akt. In primary saphenous vein endothelial cells (SVEC) from patients with type 2 diabetes mellitus undergoing coronary artery bypass (CABG) surgery, hybrid receptor expression was greater than in patients without type 2 diabetes mellitus. The small molecule significantly reduced hybrid expression in SVEC from patients with type 2 diabetes mellitus.

Conclusions

We identified a small molecule that decreases the formation of IR: IGF-1R hybrid receptors in human endothelial cells, without significant impact on the overall expression of IR or IGF-1R. In HUVECs, reduction of IR: IGF-1R hybrid receptors leads to an increase in insulin-induced serine phosphorylation of the critical downstream signalling kinase, Akt. The underpinning mechanism appears, at least in part to involve the attenuation of the inhibitory effect of IR: IGF-1R hybrid receptors on PI3-kinase signalling.

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小分子调节人内皮细胞中的胰岛素受体-胰岛素样生长因子-1 受体异二聚体。

目的：胰岛素受体（IR）和胰岛素样生长因子-1 受体（IGF-1R）是由两个胞外 α 亚基和两个跨膜 β 亚基组成的异二聚体。胰岛素 αβ 和胰岛素样生长因子-1 αβ 半受体可以异源二聚体形成由一个 IR αβ 和一个 IGF-1R αβ 组成的混合物。混合体在内皮中的功能尚不清楚。我们开发了一种能够减少内皮细胞中杂交形成的小分子，以寻求深入了解：我们根据apo杂交结构的同源模型进行了高通量小分子筛选。方法：我们根据载脂蛋白杂交结构同源模型进行了高通量小分子筛选，并使用 Western 印迹法和 qPCR 对内皮细胞进行了研究，以确定能减少杂交形成的小分子的作用：我们的研究揭示了一种首创的含喹啉杂环小分子，它能将人脐静脉内皮细胞（HUVECs）中的杂合体减少 50%以上，而对 IR 或 IGF-1R 没有影响。这种小分子能减少磷脂酰肌醇 3- 激酶负调控 p85α 亚基的表达，增加下游靶点 Akt 的基础磷酸化，并增强胰岛素/胰岛素样生长因子-1 和剪切应力诱导的 Akt 丝氨酸磷酸化。在接受冠状动脉搭桥术（CABG）的 2 型糖尿病患者的原代隐静脉内皮细胞（SVEC）中，杂交受体的表达高于未接受冠状动脉搭桥术（CABG）的 2 型糖尿病患者。该小分子能明显降低 2 型糖尿病患者 SVEC 中杂交受体的表达：我们发现了一种能减少人内皮细胞中IR：IGF-1R混合受体形成的小分子，但对IR或IGF-1R的整体表达无明显影响。在 HUVECs 中，IR：IGF-1R 混合受体的减少导致胰岛素诱导的关键下游信号激酶 Akt 的丝氨酸磷酸化增加。其基本机制似乎至少部分涉及减弱IR：IGF-1R混合受体对PI3-激酶信号传导的抑制作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular and Cellular Endocrinology 医学-内分泌学与代谢

CiteScore

9.00

自引率

2.40%

发文量

174

审稿时长

42 days

期刊介绍： Molecular and Cellular Endocrinology was established in 1974 to meet the demand for integrated publication on all aspects related to the genetic and biochemical effects, synthesis and secretions of extracellular signals (hormones, neurotransmitters, etc.) and to the understanding of cellular regulatory mechanisms involved in hormonal control.