White Bobby, Wang Zhenyi, Dean Matthew, Michl Johanna, Nieora Natalia, Flannery Sarah, Vendrell Iolanda, Fischer Roman, Hulikova Alzbeta, Swietach Pawel
Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK.
Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford , Oxford, UK.
J Cell Biol. 2025 Aug 4;224(8). doi: 10.1083/jcb.202409103. Epub 2025 Jun 24.
Hypoxia-inducible factors (HIFs) mediate cellular responses to low oxygen, notably enhanced fermentation that acidifies poorly perfused tissues and may eventually become more damaging than adaptive. How pH feeds back on hypoxic signaling is unclear but critical to investigate because acidosis and hypoxia are mechanistically coupled in diffusion-limited settings, such as tumors. Here, we examined the pH sensitivity of hypoxic signaling in colorectal cancer cells that can survive acidosis. HIF-1α stabilization under acidotic hypoxia was transient, declining over 48 h. Proteomic analyses identified responses that followed HIF-1α, including canonical HIF targets (e.g., CA9, PDK1), but these did not reflect a proteome-wide downregulation. Enrichment analyses suggested a role for lysosomal degradation. Indeed, HIF-1α destabilization was blocked by inactivating lysosomes, but not proteasome inhibitors. Acidotic hypoxia stimulated lysosomal activity and autophagy via mammalian target of rapamycin complex I (mTORC1), resulting in HIF-1α degradation. This response protects cells from excessive acidification by unchecked fermentation. Thus, alkaline conditions are permissive for at least some aspects of HIF-1α signaling.
缺氧诱导因子(HIFs)介导细胞对低氧的反应,特别是增强发酵,这会使灌注不良的组织酸化,最终可能比适应性反应更具破坏性。目前尚不清楚pH值如何反馈低氧信号,但由于酸中毒和缺氧在扩散受限的环境(如肿瘤)中存在机制上的联系,因此对其进行研究至关重要。在这里,我们研究了能够在酸中毒环境中存活的结肠癌细胞中低氧信号的pH敏感性。在酸性缺氧条件下,HIF-1α的稳定是短暂的,在48小时内下降。蛋白质组学分析确定了HIF-1α之后的反应,包括典型的HIF靶点(如CA9、PDK1),但这些反应并未反映全蛋白质组范围的下调。富集分析表明溶酶体降解起作用。事实上,通过使溶酶体失活可阻止HIF-1α的不稳定,但蛋白酶体抑制剂则无此作用。酸性缺氧通过雷帕霉素复合物I(mTORC1)刺激溶酶体活性和自噬,导致HIF-1α降解。这种反应可保护细胞免受不受控制的发酵导致的过度酸化。因此,碱性条件至少对HIF-1α信号传导的某些方面是允许的。