Complement C3a activates osteoclasts by regulating the PI3K/PDK1/SGK3 pathway in patients with multiple myeloma.
Complement C3a activates osteoclasts by regulating the PI3K/PDK1/SGK3 pathway in patients with multiple myeloma.
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补体 C3a 通过调节多发性骨髓瘤患者的 PI3K/PDK1/SGK3 通路激活破骨细胞
DOI：
10.20892/j.issn.2095-3941.2020.0430
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发表时间：
2021-05-07
影响因子：
5.5
通讯作者：
Fu R
中科院分区：
文献类型：
--
作者：
Jiang F;Liu H;Peng F;Liu Z;Ding K;Song J;Li L;Chen J;Shao Q;Yan S;De Veirman K;Vanderkerken K;Fu R
目的：骨髓瘤骨病（MBD）是多发性骨髓瘤（MM）最常见的并发症。我们前期的研究表明MM患者血清C3/C4水平与骨病的严重程度呈显著正相关。然而，C3 a/C4 a在破骨细胞MM患者中的机制仍不清楚。方法：体外实验观察C3 a/C4 a对破骨细胞形成和功能的影响。利用RNA-seq分析筛选影响破骨细胞的潜在通路，并通过Western blot、qRT-PCR和通路抑制剂对结果进行验证。结果如下：与对照组（0 μg/mL）（34.635 ± 8.916%）相比，1 μg/mL和10 μg/mL C3 a诱导的破骨细胞面积/视野（mean ± SD：50.828 ± 12.984%）（53.663 ± 12.685%）显著增加（分别为P < 0.001和P < 0.001）。1 μg/mL诱导的基因OSCAR/TRAP/RANKL/组织蛋白酶K的相对mRNA表达（中位数：分别为5.041、3.726、1.638和4.752）和10 μg/mL与对照组相比，C3 a（中位数：分别为5.140、3.702、2.250和5.172）显著升高（中位数：分别为3.137、2.004、0.573和2.257）（分别为1 μg/mL P = 0.001、P = 0.003、P < 0.001和P = 0.008;分别为10 μg/mL：P < 0.001、P = 0.019、P < 0.001和P = 0.002）。1 μg/mL和10 μg/mL的C3 a诱导的破骨细胞吸收陷窝的吸收面积（平均值±标准差：51.464 ± 11.983%）和（50.219 ± 12.067%）也显著增加（33.845 ± 8.331%）（分别为P < 0.001和P < 0.001）。C4 a和对照组之间没有差异。RNA-seq分析显示C3 a通过磷酸肌醇3-激酶（PI 3 K）信号通路促进破骨细胞增殖。C3 a组PIK 3CA/磷酸肌醇依赖性激酶1（PDK 1）/血清和糖皮质激素诱导蛋白激酶3（SGK 3）基因及PI 3 K/PDK 1/p-SGK 3蛋白的相对表达均显著高于对照组。SGK抑制剂（EMD 638683）降低了MM患者破骨细胞中C3 a的活化作用。结论：C3 a通过调节MM患者的PI 3 K/PDK 1/SGK 3通路激活破骨细胞，使用SGK抑制剂可降低该通路。总体而言，我们的研究结果确定了MBD患者的潜在治疗靶点和策略。
Objective: Myeloma bone disease (MBD) is the most common complication of multiple myeloma (MM). Our previous study showed that the serum levels of C3/C4 in MM patients were significantly positively correlated with the severity of bone disease. However, the mechanism of C3a/C4a in osteoclasts MM patients remains unclear. Methods: The formation and function of osteoclasts were analyzed after adding C3a/C4a in vitro. RNA-seq analysis was used to screen the potential pathways affecting osteoclasts, and the results were verified by Western blot, qRT-PCR, and pathway inhibitors. Results: The osteoclast area per view induced by 1 μg/mL (mean ± SD: 50.828 ± 12.984%) and 10 μg/mL (53.663 ± 12.685%) of C3a was significantly increased compared to the control group (0 μg/mL) (34.635 ± 8.916%) (P < 0.001 and P < 0.001, respectively). The relative mRNA expressions of genes, OSCAR/TRAP/RANKL/cathepsin K, induced by 1 μg/mL (median: 5.041, 3.726, 1.638, and 4.752, respectively) and 10 μg/mL (median: 5.140, 3.702, 2.250, and 5.172, respectively) of C3a was significantly increased compared to the control group (median: 3.137, 2.004, 0.573, and 2.257, respectively) (1 μg/mL P = 0.001, P = 0.003, P < 0.001, and P = 0.008, respectively; 10 μg/mL: P < 0.001, P = 0.019, P < 0.001, and P = 0.002, respectively). The absorption areas of the osteoclast resorption pits per view induced by 1 μg/mL (mean ± SD: 51.464 ± 11.983%) and 10 μg/mL (50.219 ± 12.067%) of C3a was also significantly increased (33.845 ± 8.331%) (P < 0.001 and P < 0.001, respectively) compared to the control. There was no difference between the C4a and control groups. RNA-seq analysis showed that C3a promoted the proliferation of osteoclasts using the phosphoinositide 3-kinase (PI3K) signaling pathway. The relative expressions of PIK3CA/phosphoinositide dependent kinase-1 (PDK1)/serum and glucocorticoid inducible protein kinases (SGK3) genes and PI3K/PDK1/p-SGK3 protein in the C3a group were significantly higher than in the control group. The activation role of C3a in osteoclasts of MM patients was reduced by the SGK inhibitor (EMD638683). Conclusions: C3a activated osteoclasts by regulating the PI3K/PDK1/SGK3 pathways in MM patients, which was reduced using a SGK inhibitor. Overall, our results identified potential therapeutic targets and strategies for MBD patients.
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影响因子：
5.3
作者：
Jiang F;Liu H;Liu Z;Yan S;Chen J;Shao Q;Li L;Song J;Wang G;Shao Z;Fu R
通讯作者：
Fu R
影响因子：
--
作者：
Oranger A;Carbone C;Izzo M;Grano M
通讯作者：
Grano M
影响因子：
5.4
作者：
Liu Z;Jing Q;Wang Y;Li Y;Mi F;Xiang C;Fu R
通讯作者：
Fu R
影响因子：
3.7
作者：
Andrades, JA;Nimni, ME;Sorgente, N
通讯作者：
Sorgente, N
影响因子：
12.3
作者：
Langmead B;Trapnell C;Pop M;Salzberg SL
通讯作者：
Salzberg SL