低氧应激与脂质代谢通路失衡密切相关。分子伴侣Hsp90是蛋白质质量控制的重要分子，多种底物蛋白参与脂质代谢调控。HIF-1α，是Hsp90的底物蛋白，亦是参与脂质代谢通路调控的重要因子，同时与AMPK存在协同调控，但低氧应激下Hsp90对其协同调控机制亟待阐明。本课题组前期研究结果表明Hsp90α有稳定转录因子BCLAF1作用；低氧应激下BCLAF1对HIF-1α存在转录调控；Hsp90抑制导致低氧应激下脂质合成减少。本课题将BCLAF1作为底物蛋白，以CRISPR-Cas9技术建立Hsp90敲除及表达Hsp90ATPase 突变体和不同结构域的细胞株，应用免疫共沉淀、DUO-LINK技术分析低氧应激下Hsp90-BCLAF1间相互作用，及对HIF-1α、AMPK及其下游脂质通路、及脂质组学的影响，阐明低氧应激中Hsp90-BCLAF1-HIF1α/AMPK轴相关的脂质代谢通路调控机制。

Hypoxia induces lipid metabolism dyshomeostasis. Molecular chaperone Hsp90 is important for maintaining protein quality control, and various client proteins of Hsp90 plays important roles in lipid metabolism.HIF-1α,a client protein of Hsp90, is an critical regulator of lipid metabolism, and has a complicated feedback regulation with AMPK.The role of Hsp90 in synergistically regulating HIF-1α/AMPK related lipid metabolism pathway remains unclear.Our data suggest Hsp90 stabilized the transcription regulator BCLAF1,BCLAF1 regulated the transcription of HIF1α, inhibition of Hsp90 induced the inhibition of the synthesis of fatty acid.The study will use BCLAF1 as a substrate protein, establish Hsp90 knockout and Hsp90 mutants expressed cell line with CRISPR-Cas9 technique, with the help of coimmunoprecipitation and Duo-Link analysis the interaction of Hsp90-BCLAF1 under hypoxia.With the help of RNAseq and ChIP, analysis the effect of Hsp90 and BCLAF1 on downstream lipid metabolism pathway adn lipidomics and explore the regulation mechanism of the axis of Hsp90-BCLAF1-HIF-1α/AMPK in lipid metabolism.