ZLN005通过PGC-1α调控糖酵解及线粒体代谢诱导多发性骨髓瘤凋亡及其机制研究

Aberrant aerobic glycolysis and mitochondrial metabolism are important features of multiple myeloma(MM) metabolism, and closely correlated with efficacy and prognosis of MM. As a regulator of metabolism, ZLN005 regulates metabolism of normal tissues and cells, however, its effect on tumor metabolism has not been reported. Our preliminary study found that ZLN005 could effectively inhibit the sugar, lactic acid and glutamine metabolism of MM, reduce expression of peroxisome proliferator-activated receptor γ coactivator-1 α(PGC-1α) and significantly induce apoptosis of MM cells. Meanwhile, our previous studies have revealed that PGC-1α play important roles in regulation of MM abnormal metabolisms. Accordingly, we speculate that ZLN005 regulates glycolysis and mitochondrial metabolism via targeting PGC-1α, leading to apoptosis of MM. Next, we plan to evaluate the impact of ZLN005 on abnormal metabolism and apoptosis in MM by analyzing clinical samples and performing in-vitro and in-vivo studies, to study whether ZLN005 exerts its effects on mitochondrial metabolism and glycolysis via regulation of PGC-1α by CRISPR/Cas9 gene editing technology, aiming to demonstrate that the impaired metabolism and apoptosis of MM cells caused by ZLN005 is realized by PGC-1α regulated expressions of several key factors of metabolism pathways, and the feasibility and possible mechanism of ZLN005 targeting metabolism to improve efficacy and prognosis of MM will finally be evaluated. Our work will provide a new treatment strategy for treating MM.

异常糖酵解与线粒体代谢是多发性骨髓瘤（MM）重要代谢特点，与疗效预后密切相关。ZLN005是一种代谢调节药物，调控正常组织代谢，但对肿瘤代谢有何影响，目前尚无报道。本课题预实验发现ZLN005可显著抑制MM糖、乳酸及谷氨酰胺代谢，降低过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)表达，并诱导MM细胞凋亡。而本课题前期研究发现PGC-1α是调控MM异常代谢的关键因子。据此推测ZLN005通过PGC-1α调控糖酵解及线粒体代谢，导致MM凋亡。本课题拟结合临床资料及体内外实验分析ZLN005对MM异常代谢及凋亡的影响，采用CRISPR/Cas9等基因技术研究ZLN005是否通过PGC-1α对线粒体代谢及糖酵解发挥调控，证实ZLN005通过PGC-1α调控多个代谢通路关键因子影响代谢并导致MM凋亡，探讨ZLN005干预代谢实现改善MM疗效、预后的可行性及潜在机制，为MM治疗提供新策略

异常糖酵解与线粒体代谢是多发性骨髓瘤（MM）重要代谢特点，与疗效预后密切相关。本课题前期研究发现过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)是调控MM异常代谢的关键因子。ZLN005是一种代谢调节药物，调控正常组织代谢，但对肿瘤代谢有何影响，目前报道较少。本课题推测ZLN005可显著抑制MM糖、乳酸及谷氨酰胺代谢，降低PGC-1α表达，并诱导MM细胞凋亡。本课题通过体外实验分析了ZLN005对MM异常代谢及凋亡的影响，发现ZLN005处理后的RPMI8226细胞凋亡水平较对照组显著升高，药物处理组的细胞增殖受到抑制并阻滞于G2/M期，药物组细胞伴随着葡萄糖、谷氨酰胺消耗和乳酸生成的减缓，线粒体排列紊乱，形态肿胀，线粒体发生自噬。进一步研究发现ZLN005可降低MM细胞的线粒体膜电位，下调线粒体呼吸链复合物I活力，抑制活性氧生成，减少ATP的合成。基因芯片结果发现有1147个基因上调，786个基因下调，富集分析发现ZLN005明显影响细胞凋亡相关通路、细胞周期相关通路、代谢相关通路、PI3K/Akt信号通路、MAPK信号通路等；Western-blotting法检测显示经ZLN005处理RPMI8226细胞24h后BAX水平显著增高，PGC-1α、BCL-2、mtTFA、LDHA、GLUT1水平显著降低，差异有统计学意义（p＜0.05）。随后的体内实验发现ZLN005处理后的肿瘤组织生长缓慢，体积显著小于对照组（p＜0.05），但体重与对照组无显著差异。为进一步验证结果，采用质粒转染等基因技术研究ZLN005通过PGC-1α对线粒体代谢及糖酵解发挥调控，并证实了ZLN005通过PGC-1α调控多个代谢通路关键因子影响代谢并导致MM凋亡，为ZLN005干预代谢实现改善MM疗效、预后的可行性提供了理论依据，为MM的治疗提供了新策略。

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