我们最近发表了FDA批准的乳腺癌药物palbociclib（CDK4/6抑制剂）的热蛋白组分析（TPP）。我们最近还通过体内成像、代谢组学和RNaseq鉴定了小鼠肝再生过程中的代谢变化。我现在提议用TPP方法研究人类细胞和小鼠肝再生过程中的新陈代谢。初步实验表明，某些蛋白质的热稳定性反映了细胞代谢。为了全面分析蛋白质组结构的变化，我建议研究培养人细胞在八种代谢状态下蛋白质组的热稳定性变化。我们还将通过研究磷酸化和乙酰化对人体蛋白质组热稳定性的影响，进一步发展该方法。最后，我们将小鼠肝蛋白质组在肝再生过程中的热稳定性数据与以前的代谢组学和RNaseq数据进行了比较和整合。热蛋白质组分析的应用有望提供以前无法获得的细胞代谢状态的见解。这对于新陈代谢的基础研究，包括癌症新陈代谢、肥胖和糖尿病，都是有意义的。我们的研究结果也可能对基于机制的蛋白质生物标志物的开发具有临床应用价值。

Metabolic diseases including obesity and diabetes are becoming more and more common in China. Better understanding of cellular metabolism is critical for diagnosis and treatment of both acquired and inherited metabolic diseases. It is now known that drug binding inside cells induces thermal stability changes of target proteins. This allows label-free analysis of drug binding in cells by mass spectrometry based “thermal proteome profiling” (TPP). Despite the great promise of this technology for drug binding, biomarker development and other applications, TPP methodology has not been widely adopted in China..We recently published thermal proteome profiling of FDA approved breast cancer drug palbociclib (CDK4/6 inhibitor). We also recently identified metabolic changes during mouse liver regeneration using in vivo imaging, metabolomics and RNAseq. I now propose to study metabolism in human cells and during mouse liver regeneration using TPP methodology. Preliminary experiments indicate that thermal stability of some proteins reflects cellular metabolism. Furthermore, mitochondrial metabolism is closely associated with phosphorylation and acetylation, but most of these identified post-translational modifications do not have any known functional role..To comprehensively analyze changes in proteome organization, I propose to study thermal stability changes of the proteome under eight metabolic states in cultured human cells. We will also further develop the methodology by studying the effect of phosphorylation and acetylation for thermal stability of the human proteome. Finally, we compare and integrate thermal stability data of mouse liver proteome during liver regeneration with our previous metabolomics and RNAseq data..In summary, application of thermal proteome profiling is expected to provide previously unattainable insights into cellular metabolic state could be transformative for basic research on metabolism, including cancer metabolism, obesity and diabetes. Our findings may have clinical utility for the development of mechanism-based protein biomarkers.