Bay11-7082纳米胶束通过CARD8抑制新型髓核祖细胞焦亡再生退变椎间盘的双元调控机制

髓核细胞消亡是DDD关键致病因素，聚焦种子细胞和改善其凋亡微环境的前沿研究及预实验提示：焦亡微环境是被忽视的制约细胞治疗疗效的“盲区”。申请人研发了新型髓核祖细胞(NPPCs)库平台，发现Bay11-7082纳米胶束可通过经典炎症小体介导的快速焦亡通路24h内明显抑制了mNPPCs焦亡，而其通过CARD8介导的慢速焦亡通路48h后才显著抑制hNPPCs焦亡的种属差异现象。我们提出Bay11-7082分别通过NLRP1/ASC/pro-CASP-1/CASP-1/GSDMD/IL-1β或CARD8/pro-CASP-1/GSDMD通路，抑制mNPPCs或hNPPCs焦亡的假说。本项目计划，验证Bay11-7082协同NPPCs体内再生退变椎间盘的双元疗效，结合基因编辑NPPCs模式细胞和转基因小鼠，阐明其抑制不同种属NPPCs焦亡的具体机制。验证生物活性载体协同种子细胞双元再生DDD的策略。

The degeneration of nucleus pulposus (NP) is not only the main cause of degenerative disc disease (DDD), but also considered to be its initial factor. One frontier focus of research is to find optimum seed cells and improve its apoptotic microenvironment, and our preliminary experiment indicated that pyroptosis microenvironment severely dampens the effect of cell therapy but largely neglected as a "blind spot" of research. We have established a new nucleus pulposus progenitor cell (NPPC) line, and intriguingly observed a species-specific distinction. We found that the Bay11-7082 nano-micelles significantly inhibited mNPPCs pyroptosis within 24 hours of treatment by targeting the inflammasome-mediated classical rapid pyroptosis pathway, while it could significantly inhibit hNPPCs pyroptosis only after 48 hours via the Caspase Recruitment Domain Family Member 8 (CARD8) mediated slow pyroptosis pathway. We propose that Bay11-7082 inhibits the pyroptosis of mNPPCs by modulating NLRP1/ASC/ pro-CASP-1 / CASP-1 /GSDMD/ Il-1β pathway and CARD8/ pro-CASP-1 /GSDMD pathway in hNPPCs, respectively. This project plans to verify the efficacy of Bay11-7082 in synergy with NPPCs for intervertebral disc regeneration in vivo, and employ both gene-edited NPPCs cells and transgenic mice to explore the underlying mechanism of inhibiting the caustic death of different NPPCs species. It is expected to provide new research model cells and shed light on the prevention and treatment of DDD.