白细胞介素-32受体的鉴定及其信号通路的研究

Interleukin-32 (IL-32) is a pro-inflammatory cytokine. It is mainly secreted by activated immune cells and highly expressed in immune tissues. IL-32 efficiently induces various cytokines such as TNFα, IFNβ and IL-1 in many cells, especially in monocytes, macrophages, and PBMCs, by activating NF-kB and MAPK signaling pathways. Since a large number of studies have demonstrated that IL-32 is associated with numerous diseases, ranging from infectious diseases, inflammatory diseases and cancer, it would be of great importance to develop targeting strategies for IL-32. However, an unambiguous cell surface receptor for IL-32 that would transmit a signal has not been identified, and the precise signaling pathway downstream of IL-32 is still unknown. Here we will use the newly developed SWATH-MS method to analyze the purified IL-32 protein complex to get the candidate proteins. Diversified approaches such as co-immunoprecipitation, subcellular localization analysis and gene editing would be used to identify the cell surface receptor for IL-32 and the key signaling molecules downstream of IL-32. The molecular mechanism by which these novel proteins participate in IL-32 signaling pathway will also be studies. We believe that this study will lead to great progress in the understanding of the molecular mechanism of IL-32 signaling pathway and the clinical application of IL-32 in various disease processes.

白细胞介素-32（Interleukin-32，IL-32）是一种促炎性细胞因子，主要是由免疫细胞分泌，在免疫器官中大量表达，可以激活多种免疫细胞的NF-κB和MAPK信号通路，进而诱导产生TNFα，IL-1等细胞因子。研究表明IL-32与许多疾病密切相关，包括炎症性疾病，感染性疾病和癌症等，在临床上有极大的应用前景。然而IL-32的细胞膜表面受体尚未被鉴定，其信号转导的核心途径也未被阐明，这极大地限制了IL-32的临床应用。本项目拟利用最新的SWATH-MS技术对纯化的IL-32复合体进行定性定量分析，得到候选蛋白，再通过相互作用分析、细胞定位分析、基因敲除检测生物学功能等方法进行验证，从而鉴定出IL-32的细胞膜表面受体及其信号转导过程中的关键蛋白，并对其作用机理进行初步探究。本项目的完成将完善IL-32信号通路的基础研究，为IL-32的临床应用提供重要的理论依据。

白细胞介素-32 （Interleukin-32, IL-32）是一种主要由免疫细胞分泌的促炎性细胞因子。它可以有效地激活多种免疫细胞的NF-κB和MAPK信号通路，诱导产生TNFα, IL-1等细胞因子，进而参与到多种免疫性疾病中。然而IL-32的细胞膜表面受体尚未被鉴定，其信号转导的核心途径也未被阐明。为鉴定IL-32的细胞膜表面受体，我们开发了最新的APEX2临近标记技术，联合定量质谱技术，鉴定出膜蛋白CD14是介导IL-32信号的受体复合物中的关键组分。我们在人源和鼠源多种细胞系统里研究发现，CD14对IL-32的信号转导必不可少。我们的研究还初步揭示了IL-32下游信号通路，发现IL-32通过TAK1-IKKβ-NF-κB信号轴诱导TNFα等细胞因子的产生。另外，我们还发现Src激酶蛋白对于IL-32信号传导也至关重要。该项目的研究鉴定了IL-32的受体成分，将极大地推动IL-32信号通路的研究，并为临床上开发针对 IL-32 的治疗方案提供重要的理论依据。联合利用APEX2临近标记和定量质谱技术鉴定膜蛋白组分的研究方法也将为同类研究提供方法学上的参考。