Clinic Center of Human Gene Research, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Ave, Wuhan, 430022, China.
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
J Transl Med. 2024 Jan 18;22(1):74. doi: 10.1186/s12967-024-04872-x.
Angiogenesis is essential for tissue repair in ischemic diseases, relying on glycolysis as its primary energy source. Prolyl 4-hydroxylase subunit alpha 1 (P4HA1), the catalytic subunit of collagen prolyl 4-hydroxylase, is a glycolysis-related gene in cancers. However, its role in glycolysis-induced angiogenesis remains unclear.
P4HA1 expression was modulated using adenoviruses. Endothelial angiogenesis was evaluated through 5-ethynyl-2'-deoxyuridine incorporation, transwell migration, and tube formation assays in vitro. In vivo experiments measured blood flow and capillary density in the hindlimb ischemia (HLI) model. Glycolytic stress assays, glucose uptake, lactate production, and quantitative reverse transcription-polymerase chain reaction (RT-PCR) were employed to assess glycolytic capacity. Transcriptome sequencing, validated by western blotting and RT-PCR, was utilized to determine underlying mechanisms.
P4HA1 was upregulated in endothelial cells under hypoxia and in the HLI model. P4HA1 overexpression promoted angiogenesis in vitro and in vivo, while its knockdown had the opposite effect. P4HA1 overexpression reduced cellular α-ketoglutarate (α-KG) levels by consuming α-KG during collagen hydroxylation. Downregulation of α-KG reduced the protein level of a DNA dioxygenase, ten-eleven translocation 2 (TET2), and its recruitment to the fructose-1,6-biphosphatase (FBP1) promoter, resulting in decreased FBP1 expression. The decrease in FBP1 enhanced glycolytic metabolism, thereby promoting endothelial angiogenesis.
Hypoxia-induced endothelial P4HA1 overexpression enhanced angiogenesis by promoting glycolytic metabolism reprogramming through the P4HA1/α-KG/TET2/FBP1 pathway. The study's findings underscore the significance of P4HA1 in post-ischemic angiogenesis, suggesting its therapeutic potential for post-ischemic tissue repair.
血管生成对于缺血性疾病中的组织修复至关重要，其主要依赖糖酵解作为能量来源。脯氨酰 4-羟化酶亚基 α1（P4HA1）是胶原脯氨酰 4-羟化酶的催化亚基，是癌症中与糖酵解相关的基因。然而，其在糖酵解诱导的血管生成中的作用尚不清楚。
使用腺病毒调节 P4HA1 的表达。通过 5-乙炔基-2'-脱氧尿苷掺入、体外 Transwell 迁移和管形成实验评估内皮血管生成。在肢体缺血（HLI）模型中测量血流和毛细血管密度的体内实验。通过糖酵解应激测定、葡萄糖摄取、乳酸生成和定量逆转录聚合酶链反应（RT-PCR）评估糖酵解能力。通过 Western 印迹和 RT-PCR 验证进行转录组测序，以确定潜在的机制。
在缺氧条件下和 HLI 模型中，内皮细胞中 P4HA1 上调。P4HA1 过表达促进体外和体内血管生成，而过表达则相反。P4HA1 过表达通过在胶原羟化过程中消耗 α-KG 来降低细胞 α-酮戊二酸（α-KG）水平。α-KG 的下调降低了 DNA 双加氧酶 ten-eleven 易位 2（TET2）的蛋白水平及其在果糖-1,6-二磷酸酶（FBP1）启动子上的募集，导致 FBP1 表达降低。FBP1 的减少增强了糖酵解代谢，从而促进内皮血管生成。
缺氧诱导的内皮 P4HA1 过表达通过 P4HA1/α-KG/TET2/FBP1 途径促进糖酵解代谢重编程，从而增强血管生成。该研究结果强调了 P4HA1 在缺血后血管生成中的重要性，表明其在缺血后组织修复中的治疗潜力。
