Department of Biomedical Sciences, School of Graduate Studies, Meharry Medical College, Nashville, TN 37208, USA.
Center for AIDS Health Disparities Research, Meharry Medical College, Nashville, TN 37208, USA.
Int J Mol Sci. 2024 Mar 15;25(6):3324. doi: 10.3390/ijms25063324.
Genetic variants in the protein-coding regions of APOL1 are associated with an increased risk and progression of chronic kidney disease (CKD) in African Americans. Hypoxia exacerbates CKD progression by stabilizing HIF-1α, which induces APOL1 transcription in kidney podocytes. However, the contribution of additional mediators to regulating APOL1 expression under hypoxia in podocytes is unknown. Here, we report that a transient accumulation of HIF-1α in hypoxia is sufficient to upregulate APOL1 expression in podocytes through a cGAS/STING/IRF3-independent pathway. Notably, IFI16 ablation impedes hypoxia-driven APOL1 expression despite the nuclear accumulation of HIF-1α. Co-immunoprecipitation assays indicate no direct interaction between IFI16 and HIF-1α. Our studies identify hypoxia response elements (HREs) in the APOL1 gene enhancer/promoter region, showing increased HIF-1α binding to HREs located in the APOL1 gene enhancer. Luciferase reporter assays confirm the role of these HREs in transcriptional activation. Chromatin immunoprecipitation (ChIP)-qPCR assays demonstrate that IFI16 is not recruited to HREs, and IFI16 deletion reduces HIF-1α binding to APOL1 HREs. RT-qPCR analysis indicates that IFI16 selectively affects APOL1 expression, with a negligible impact on other hypoxia-responsive genes in podocytes. These findings highlight the unique contribution of IFI16 to hypoxia-driven APOL1 gene expression and suggest alternative IFI16-dependent mechanisms regulating APOL1 gene expression under hypoxic conditions.
APOL1 蛋白编码区的遗传变异与非裔美国人慢性肾脏病 (CKD) 的风险增加和进展有关。缺氧通过稳定 HIF-1α 来加重 CKD 进展,HIF-1α 诱导肾脏足细胞中 APOL1 的转录。然而,在缺氧条件下,其他介质对调节足细胞中 APOL1 表达的贡献尚不清楚。在这里,我们报告在缺氧条件下,HIF-1α 的短暂积累足以通过 cGAS/STING/IRF3 非依赖性途径上调足细胞中 APOL1 的表达。值得注意的是,IFI16 缺失尽管 HIF-1α 核积累,但会阻碍缺氧驱动的 APOL1 表达。共免疫沉淀测定表明 IFI16 和 HIF-1α 之间没有直接相互作用。我们的研究在 APOL1 基因增强子/启动子区域中鉴定出缺氧反应元件 (HRE),显示出 HIF-1α 与位于 APOL1 基因增强子中的 HRE 结合增加。荧光素酶报告基因测定证实了这些 HRE 在转录激活中的作用。染色质免疫沉淀 (ChIP)-qPCR 测定表明 IFI16 不会被募集到 HRE 上,并且 IFI16 缺失会降低 HIF-1α 与 APOL1 HRE 的结合。RT-qPCR 分析表明,IFI16 选择性地影响 APOL1 的表达,对足细胞中其他缺氧反应基因的影响可以忽略不计。这些发现强调了 IFI16 对缺氧驱动的 APOL1 基因表达的独特贡献,并表明在缺氧条件下,IFI16 依赖的替代机制调节 APOL1 基因表达。