DataSheet1_Integrating Mendelian randomization and single-cell RNA sequencing to identify therapeutic targets of baicalin for type 2 diabetes mellitus.PDF

BackgroundAlternative and complementary therapies play an imperative role in the clinical management of Type 2 diabetes mellitus (T2DM), and exploring and utilizing natural products from a genetic perspective may yield novel insights into the mechanisms and interventions of the disorder.
MethodsTo identify the therapeutic target of baicalin for T2DM, we conducted a Mendelian randomization study. Druggable targets of baicalin were obtained by integrating multiple databases, and target-associated cis-expression quantitative trait loci (cis-eQTL) originated from the eQTLGen consortium. Summary statistics for T2DM were derived from two independent genome-wide association studies available through the DIAGRAM Consortium (74,124 cases vs. 824,006 controls) and the FinnGen R9 repository (9,978 cases vs. 12,348 controls). Network construction and enrichment analysis were applied to the therapeutic targets of baicalin. Colocalization analysis was utilized to assess the potential for the therapeutic targets and T2DM to share causative genetic variations. Molecular docking was performed to validate the potency of baicalin. Single-cell RNA sequencing was employed to seek evidence of therapeutic targets’ involvement in islet function.
ResultsEight baicalin-related targets proved to be significant in the discovery and validation cohorts. Genetic evidence indicated the expression of ANPEP, BECN1, HNF1A, and ST6GAL1 increased the risk of T2DM, and the expression of PGF, RXRA, SREBF1, and USP7 decreased the risk of T2DM. In particular, SREBF1 has significant interaction properties with other therapeutic targets and is supported by strong colocalization. Baicalin had favorable combination activity with eight therapeutic targets. The expression patterns of the therapeutic targets were characterized in cellular clusters of pancreatic tissues that exhibited a pseudo-temporal dependence on islet cell formation and development.
ConclusionThis study identified eight potential targets of baicalin for treating T2DM from a genetic perspective, contributing an innovative analytical framework for the development of natural products. We have offered fresh insights into the connections between therapeutic targets and islet cells. Further, fundamental experiments and clinical research are warranted to delve deeper into the molecular mechanisms of T2DM.

**背景** 替代与补充疗法在2型糖尿病（Type 2 diabetes mellitus, T2DM）的临床管理中发挥着不可或缺的作用，从遗传学视角探索并开发天然产物，或可为该疾病的发病机制与干预手段带来全新认知。

**方法** 为明确黄芩苷（baicalin）治疗2型糖尿病的作用靶点，本研究开展了孟德尔随机化（Mendelian randomization）研究。通过整合多数据库获取黄芩苷的可药用靶点，并从eQTLGen联盟中获取与靶点相关的顺式表达数量性状位点（cis-expression quantitative trait loci, cis-eQTL）。2型糖尿病的汇总统计数据来源于两项独立的全基因组关联研究：DIAGRAM联盟（74,124例病例vs 824,006例对照）与FinnGen R9数据库（9,978例病例vs 12,348例对照）。对黄芩苷的治疗靶点进行网络构建与富集分析。采用共定位分析评估治疗靶点与2型糖尿病共享致病遗传变异的可能性。通过分子对接（molecular docking）验证黄芩苷的结合活性。利用单细胞RNA测序（single-cell RNA sequencing）探寻治疗靶点参与胰岛功能调控的相关证据。

**结果** 本研究在发现队列与验证队列中均鉴定出8个与黄芩苷相关的显著性靶点。遗传学证据显示，ANPEP、BECN1、HNF1A及ST6GAL1的高表达会增加2型糖尿病的发病风险，而PGF、RXRA、SREBF1及USP7的高表达则会降低该疾病的发病风险。其中，SREBF1与其他治疗靶点存在显著的相互作用，且得到了强有力的共定位分析结果支持。黄芩苷与上述8个治疗靶点均表现出良好的结合活性。在胰腺组织细胞簇中，治疗靶点的表达模式呈现出伪时间依赖性的胰岛细胞形成与发育特征。

**结论** 本研究从遗传学视角鉴定出8个黄芩苷治疗2型糖尿病的潜在靶点，为天然产物的开发提供了创新性的分析框架。本研究为治疗靶点与胰岛细胞之间的关联提供了全新的认知。未来需开展基础实验与临床研究，以进一步阐明2型糖尿病的分子作用机制。