School of Medicine, IMPACT, Institute for Innovation in Physical and Mental Health and Clinical Translation, Deakin University, Geelong, Australia.
School of Medicine, Bioinformatics Core Research Facility, Deakin University, Geelong, Australia.
Islets. 2023 Dec 31;15(1):2165368. doi: 10.1080/19382014.2023.2165368.
Targeting β-cell failure could prevent, delay or even partially reverse Type 2 diabetes. However, development of such drugs is limited as the molecular pathogenesis is complex and incompletely understood. Further, while β-cell failure can be modeled experimentally, only some of the molecular changes will be pathogenic. Therefore, we used a novel approach to identify molecular pathways that are not only changed in a diabetes-like state but also are reversible and can be targeted by drugs. INS1E cells were cultured in high glucose (HG, 20 mM) for 72 h or HG for an initial 24 h followed by drug addition (exendin-4, metformin and sodium salicylate) for the remaining 48 h. RNAseq (Illumina TruSeq), gene set enrichment analysis (GSEA) and pathway analysis (using Broad Institute, Reactome, KEGG and Biocarta platforms) were used to identify changes in molecular pathways. HG decreased function and increased apoptosis in INS1E cells with drugs partially reversing these effects. HG resulted in upregulation of 109 pathways while drug treatment downregulated 44 pathways with 21 pathways in common. Interestingly, while hyperglycemia extensively upregulated metabolic pathways, they were not altered with drug treatment, rather pathways involved in the cell cycle featured more heavily. GSEA for hyperglycemia identified many known pathways validating the applicability of our cell model to human disease. However, only a fraction of these pathways were downregulated with drug treatment, highlighting the importance of considering druggable pathways. Overall, this provides a powerful approach and resource for identifying appropriate targets for the development of β-cell drugs.
针对β细胞衰竭可能预防、延迟甚至部分逆转 2 型糖尿病。然而，由于发病机制复杂且尚未完全了解，此类药物的开发受到限制。此外，虽然可以通过实验模拟β细胞衰竭，但只有一些分子变化才具有致病性。因此，我们采用了一种新方法来识别不仅在类似糖尿病状态下发生变化，而且还可以通过药物逆转和靶向的分子途径。INS1E 细胞在高葡萄糖（HG，20 mM）中培养 72 小时或在 HG 中培养 24 小时，然后加入药物（exendin-4、二甲双胍和水杨酸钠）48 小时。使用 RNAseq（Illumina TruSeq）、基因集富集分析（GSEA）和途径分析（使用 Broad Institute、Reactome、KEGG 和 Biocarta 平台）来识别分子途径的变化。HG 降低了 INS1E 细胞的功能并增加了细胞凋亡，而药物部分逆转了这些作用。HG 导致 109 条途径上调，而药物治疗下调了 44 条途径，其中 21 条途径是共同的。有趣的是，虽然高血糖广泛上调代谢途径，但药物治疗并没有改变这些途径，而是更多地涉及细胞周期途径。高血糖的 GSEA 鉴定了许多已知的途径，验证了我们的细胞模型对人类疾病的适用性。然而，只有一部分途径在药物治疗后下调，这突出了考虑可用药途径的重要性。总体而言，这为识别β细胞药物开发的合适靶点提供了一种强大的方法和资源。
