Single-cell RNA sequencing identifies endothelial-derived HBEGF as promoting pancreatic beta cell proliferation in mice via the EGFR–Kmt5a–H4K20me pathway

IF 8.4 1区 医学 Q1 ENDOCRINOLOGY & METABOLISM Diabetologia Pub Date : 2024-12-19 DOI:10.1007/s00125-024-06341-y
Fengling Lai, Kaixin Zhou, Yingjie Ma, Hao Lv, Weilin Wang, Rundong Wang, Tao Xu, Rong Huang
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Abstract

Aims/hypothesis

Pancreatic beta cell mass is dynamically regulated in response to increased physiological and pathological demands. Understanding the mechanisms that control physiological beta cell proliferation could provide valuable insights into novel therapeutic approaches to diabetes. Here, we aimed to analyse the intracellular and extracellular signalling pathways involved in regulating the physiological proliferation of beta cells using single-cell RNA-seq (scRNA-seq) and in vitro functional assays.

Methods

Islets isolated from nulliparous mice, mice at different time points of gestation and mice at day 4 after delivery were analysed using scRNA-seq. Bioinformatics analyses of scRNA-seq data were performed to determine the heterogeneous transcriptomic characteristics of beta cells and to identify the proliferating subpopulation. CellChat was used to analyse cell–cell communication and identify the ligand–receptor pairs between beta cell subclusters as well as between non-beta cells and proliferating beta cells. In vitro functional assays were conducted in mouse and rat beta cell lines and isolated mouse primary islets to validate the role of Kmt5a– mono-methylation of histone H4 at lysine 20 (H4K20me) signalling and endothelial-derived heparin-binding EGF-like growth factor (HBEGF) in beta cell proliferation.

Results

Of 43,724 endocrine and non-endocrine cells within islets analysed by scRNA-seq, 15,569 beta cells were clustered into eight distinct populations, each exhibiting unique heterogeneity. A proliferating beta cell subcluster was identified that highly expressed the histone methyltransferase Kmt5a. Activation of Kmt5a–H4K20me signalling upregulated the expression of Cdk1 and promoted beta cell proliferation. The crosstalk between endothelial cells and the proliferating beta cell subcluster, mediated by the HBEGF–EGF receptor (EGFR) ligand–receptor interaction, increased as beta cell mass expanded. HBEGF increased the expression levels of genes involved in the cell cycle and promoted beta cell proliferation by regulating the Kmt5a–H4K20me signalling pathway.

Conclusions/interpretation

Our study demonstrates that, under physiological conditions, endothelial-derived HBEGF regulates beta cell proliferation through the Kmt5a–H4K20me signalling pathway, which may serve as a potential target to promote beta cell expansion and treat diabetes.

Data availability

The scRNA-seq and RNA-seq datasets are available from the Gene Expression Omnibus (GEO) using the accession numbers GSE278860 and GSE278861, respectively.

Graphical Abstract

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目的/假设胰腺β细胞的数量随着生理和病理需求的增加而动态调节。了解控制β细胞生理性增殖的机制可为糖尿病的新型治疗方法提供有价值的见解。在此,我们旨在利用单细胞RNA-seq(scRNA-seq)和体外功能测试分析参与调控β细胞生理性增殖的细胞内和细胞外信号通路。对scRNA-seq数据进行生物信息学分析,以确定β细胞的异质性转录组特征,并识别增殖亚群。CellChat 用于分析细胞-细胞间的通讯,并确定β细胞亚群之间以及非β细胞和增殖β细胞之间的配体-受体对。在小鼠和大鼠 beta 细胞系以及分离的小鼠原代胰岛中进行了体外功能测试,以验证 Kmt5a-组蛋白 H4 在赖氨酸 20 处的单甲基化 (H4K20me) 信号和内皮衍生肝素结合型 EGF 样生长因子 (HBEGF) 在 beta 细胞增殖中的作用。结果 通过scRNA-seq分析了胰岛中的43724个内分泌和非内分泌细胞,其中15569个β细胞被分为8个不同的群体,每个群体都表现出独特的异质性。发现一个增殖的β细胞亚群高度表达组蛋白甲基转移酶Kmt5a。激活Kmt5a-H4K20me信号可上调Cdk1的表达,促进β细胞增殖。由 HBEGF-EGF 受体(表皮生长因子受体)配体-受体相互作用介导的内皮细胞与增殖的β细胞亚簇之间的串扰随着β细胞体积的扩大而增加。结论/解释我们的研究表明,在生理条件下,内皮源性 HBEGF 通过 Kmt5a-H4K20me 信号通路调节β细胞增殖,这可能是促进β细胞扩增和治疗糖尿病的潜在靶点。数据提供scRNA-seq和RNA-seq数据集可从基因表达总库(GEO)获取,登录号分别为GSE278860和GSE278861。图文摘要
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来源期刊
Diabetologia
Diabetologia 医学-内分泌学与代谢
CiteScore
18.10
自引率
2.40%
发文量
193
审稿时长
1 months
期刊介绍: Diabetologia, the authoritative journal dedicated to diabetes research, holds high visibility through society membership, libraries, and social media. As the official journal of the European Association for the Study of Diabetes, it is ranked in the top quartile of the 2019 JCR Impact Factors in the Endocrinology & Metabolism category. The journal boasts dedicated and expert editorial teams committed to supporting authors throughout the peer review process.
期刊最新文献
Correction: 60th EASD Annual Meeting of the European Association for the Study of Diabetes, Madrid, Spain, 9-13 September 2024. 'Average glucose and HbA1c display a nonlinear and variable relationship: implications for clinical practice'. Glycaemic control is still central in the hierarchy of priorities in type 2 diabetes management. Lifetime history of gestational diabetes and cognitive function in parous women in midlife. Is glycaemic control still central in the hierarchy of priorities in type 2 diabetes management? The way forward is to combine glucose control and the prevention of cardiorenal complications. Exposure to antibiotics and risk of latent autoimmune diabetes in adults and type 2 diabetes: results from a Swedish case-control study (ESTRID) and the Norwegian HUNT study.
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