MIF/CD74 axis is a target for novel therapies in colon carcinomatosis.
MIF/CD74 axis is a target for novel therapies in colon carcinomatosis.
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MIF/CD74轴是结肠癌的新疗法的靶标。
DOI：
10.1186/s13046-016-0475-z
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发表时间：
2017-01-23
期刊：
影响因子：
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通讯作者：
Gariboldi M
中科院分区：
文献类型：
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作者：
Bozzi F;Mogavero A;Varinelli L;Belfiore A;Manenti G;Caccia C;Volpi CC;Beznoussenko GV;Milione M;Leoni V;Gloghini A;Mironov AA;Leo E;Pilotti S;Pierotti MA;Bongarzone I;Gariboldi M
为了提高结直肠癌腹膜癌转移（CRC-pc）患者的长期生存率，需要采取旨在获得完全细胞减灭的策略。我们从两名初治CRC患者的腹膜转移建立了类器官模型。采用标准石蜡包埋法，将其三维结构和免疫组化图谱与相应的手术标本进行比较。通过原位杂交测量CRC干细胞标志物LGR 5的RNA表达水平。通过质谱分析类器官的分泌组。4-IPP和二甲双胍干扰类器官的能量稳态。通过蛋白质印迹和质谱法研究药物治疗后的生化和代谢变化。线粒体损伤通过电子显微镜和mitotraker染色进行评估。这两个类器官在3D结构和免疫组织化学谱方面概括了其相应的临床样本，并且对癌症干细胞标志物LGR 5呈阳性。类器官的蛋白质组学分析强调了它们对能量产生途径的强烈依赖性，这表明它们的靶向可能是一种有效的治疗方法。为了验证这一假设，我们用两种靶向代谢作用于AMP活化蛋白激酶（AMPK）的药物治疗类器官，AMPK是细胞能量稳态的主要调节因子，可能在CRC中起代谢肿瘤抑制剂的作用。用4-IPP（一种激活AMPK功能的MIF/CD 74信号传导轴抑制剂）或二甲双胍（抑制线粒体呼吸链复合物I）处理类器官。作为一个新的发现，我们观察到4-IPP治疗下调AMPK信号活性，减少AKT磷酸化，激活JNK介导的应激信号反应，从而产生线粒体损伤和细胞死亡。代谢处理增强AMPK激活，降低合成代谢因子核糖体蛋白S6和p4 EBP-1的活性，并诱导线粒体去极化。我们提供的证据表明，AMPK活性的调节可能是靶向CRC-pc类器官代谢的策略。本文的在线版本（doi：10.1186/s13046-016-0475-z）包含补充材料，可供授权用户使用。
Strategies aimed at obtaining a complete cytoreduction are needed to improve long-term survival for patients with colorectal cancer peritoneal carcinomatosis (CRC-pc). We established organoid models from peritoneal metastases of two naïve CRC patients. A standard paraffin inclusion was conducted to compare their 3D structure and immunohistochemical profile with that of the corresponding surgical samples. RNA expression levels of the CRC stem cell marker LGR5 was measured by in situ hybridization. The secretome of organoids was profiled by mass spectrometry. Energy homeostasis of organoids was interfered with 4-IPP and metformin. Biochemical and metabolic changes after drug treatments were investigated by western blot and mass spectrometry. Mitochondria impairment was evaluated by electron microscopy and mitotraker staining. The two organoids recapitulated their corresponding clinical samples in terms of 3D structure and immmunoistochemical profile and were positive for the cancer stem cells marker LGR5. Proteomic analyses of organoids highlighted their strong dependence on energy producing pathways, which suggest that their targeting could be an effective therapeutic approach. To test this hypothesis, we treated organoids with two drugs that target metabolism acting on AMP-activated protein kinase (AMPK), the main regulator of cellular energy homeostasis, which may act as metabolic tumour suppressor in CRC. Organoids were treated with 4-IPP, an inhibitor of MIF/CD74 signalling axis which activates AMPK function, or metformin that inhibits mitochondrial respiratory chain complex I. As a new finding we observed that treatment with 4-IPP downregulated AMPK signalling activity, reduced AKT phosphorylation and activated a JNK-mediated stress-signalling response, thus generating mitochondrial impairment and cell death. Metformin treatment enhanced AMPK activation, decreasing the activity of the anabolic factors ribosomal protein S6 and p4EBP-1 and inducing mitochondrial depolarization. We provide evidence that the modulation of AMPK activity may be a strategy for targeting metabolism of CRC-pc organoids. The online version of this article (doi:10.1186/s13046-016-0475-z) contains supplementary material, which is available to authorized users.