GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE328nnn/GSE328627/primaryOK200TranscriptomicsHomo sapiensExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE328627GEOGSEfalseClinically approved HIF‑PHIs modulate redox metabolism, cell growth, and angiogenesis independent of HIF‑1α/HIF‑2αHIF-prolyl hydroxylase inhibitors are used to treat anemia in chronic kidney disease. These drugs stabilize hypoxia-inducible factors HIF-1α and HIF-2α, which activate erythropoiesis and iron metabolism pathways. Clinically approved HIF-PHIs including roxadustat and molidustat exhibit distinct molecular structures and selectivity profiles, yet their HIF-independent effects remain poorly understood. Here we show that roxadustat and molidustat modulate mitochondrial function, oxidative stress, lysosomal activity, and lipid accumulation, resulting in distinct cellular phenotypes in HIF-null cells. Notably, roxadustat exhibited anti-proliferative and anti-angiogenic activity in HIF-null cells, contradicting expectations of VEGF-driven angiogenesis via HIF stabilization. RNA sequencing and pathway analysis revealed compound-specific off-target gene regulation affecting cellular processes beyond canonical hypoxia responses including energy metabolism and immune signaling. These findings illuminate mechanisms underlying potential adverse effects- such as thrombosis- and identify alternative therapeutic pathways, providing a framework for optimizing HIF-PHI safety profiles and expanding their clinical applications in oncology and metabolic disorders.2026/05/21GSE328627GSM9686194GSM9686195GSM9686196GSM9686197GSM9686198GSM9686199GSM9686200GSM968620116791328627Homo sapiens[42150425]