Division of Clinical Pharmacology, Department of Molecular Pharmacology and Therapeutics, Mayo Clinic, Rochester, Minnesota.
Division of Clinical Pharmacology, Department of Molecular Pharmacology and Therapeutics, Mayo Clinic, Rochester, Minnesota
Drug Metab Dispos. 2023 Jan;51(1):1-7. doi: 10.1124/dmd.122.000945. Epub 2022 Sep 24.
Cytochrome P450s () display significant inter-individual variation in expression, much of which remains unexplained by known single-nucleotide polymorphisms (SNPs). Testis-specific Y-encoded-like proteins (s) are transcriptional regulators for several drug-metabolizing including However, transcription factors (TFs) that might influence expression through an effect on expression are unknown. Therefore, we studied regulators of expression in hepatic cell lines and their possible SNP-dependent variation. Specifically, we identified candidate TFs that might influence expression using the ENCODE ChIPseq database. Subsequently, the expression of as well as that of selected CYP targets for regulation were assayed in hepatic cell lines before and after knockdown of TFs that might influence expression through TSPYL-dependent mechanisms. Those results were confirmed by studies of TF binding to gene promoter regions. In hepatic cell lines, knockdown of the REST and ZBTB7A TFs resulted in decreased and expression and increased expression, changes reversed by overexpression. Potential binding sites for and on the promoters of and were confirmed by chromatin immunoprecipitation. Finally, common SNP variants in upstream binding sites on the promoters were identified and luciferase reporter constructs confirmed SNP-dependent modulation of gene transcription. In summary, we identified REST and ZBTB7A as regulators of the expression of TSPYL genes which themselves can contribute to regulation of expression and-potentially-of drug metabolism. SNP-dependent modulation of TSPYL transcription may contribute to individual variation in both expression and-downstream-drug response phenotypes. SIGNIFICANCE STATEMENT: Testis-specific Y-encoded-like proteins (s) are transcriptional regulators of cytochrome P450 () gene expression. Here, we report that variation in expression as a result of the effects of genetically regulated TSPYL transcription factors is an additional factor that could result in downstream variation in expression and potentially, as a result, variation in drug biotransformation.
细胞色素 P450 （CYP）在表达方面表现出显著的个体间差异，其中很大一部分仍然无法用已知的单核苷酸多态性（SNP）来解释。睾丸特异性 Y 编码样蛋白（s）是几种药物代谢酶的转录调节剂，包括 CYP。然而，可能通过影响 CYP 表达来影响 CYP 表达的转录因子（TFs）尚不清楚。因此，我们研究了肝细胞系中 CYP 表达的调节因子及其可能的 SNP 依赖性变异。具体来说，我们使用 ENCODE ChIPseq 数据库来鉴定可能影响 CYP 表达的候选 TF。随后，在敲低可能通过 TSPYL 依赖性机制影响 CYP 表达的 TF 前后，检测肝细胞系中 CYP 的表达以及 CYP 调节的选定靶标 CYP 的表达。这些结果通过研究 TF 与 CYP 基因启动子区域的结合得到证实。在肝细胞系中，REST 和 ZBTB7A TF 的敲低导致 CYP 和 CYP 表达降低，TSPYL 表达增加，这些变化通过 TSPYL 过表达逆转。CYP 和 CYP 启动子上 TSPYL 潜在结合位点通过染色质免疫沉淀得到证实。最后，确定了 CYP 启动子上游结合位点的常见 SNP 变体，并通过荧光素酶报告基因构建证实了 SNP 依赖性 CYP 基因转录的调节。总之，我们确定了 REST 和 ZBTB7A 是 TSPYL 基因表达的调节因子，而 TSPYL 基因本身可以促进 CYP 表达的调节，并且可能会调节药物代谢。TSPYL 转录的 SNP 依赖性调节可能是 CYP 表达和下游药物反应表型个体差异的另一个因素。
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