Targeted delivery of miR-99b reprograms tumor-associated macrophage phenotype leading to tumor regression.
Targeted delivery of miR-99b reprograms tumor-associated macrophage phenotype leading to tumor regression.
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miR-99b 的靶向递送可重新编程肿瘤相关巨噬细胞表型，从而导致肿瘤消退。
DOI：
10.1136/jitc-2019-000517
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发表时间：
2020-09
影响因子：
10.9
通讯作者：
Qin HY
中科院分区：
文献类型：
--
作者：
Wang L;Hu YY;Zhao JL;Huang F;Liang SQ;Dong L;Chen Y;Yu HC;Bai J;Yang JM;Fan JY;Feng L;Li SZ;Han H;Qin HY
背景越来越多的证据表明，肿瘤相关巨噬细胞(TAMs)在肿瘤进展中起着关键作用。靶向TAMS是一种潜在的肿瘤免疫治疗策略。然而，的表型和功能背后的机制还需要解决。我们以前的研究已经证明miR-125a可以逆转的抗肿瘤表型。同时，我们发现miR-125a和miR-99b聚集在同一宿主基因的第一内含子中，并在脂多糖+干扰素γ刺激下在骨髓来源的巨噬细胞中同时转录。然而，miR-99b本身是否能通过调节巨噬细胞表型发挥抗肿瘤作用尚不清楚。方法将miR-99b和/或miR-125a分别导入原位肝细胞癌(HCC)和皮下Lewis肺癌(LLC)小鼠体内。治疗效果通过活体显像、原位末端标记物染色和存活试验进行评价。采用定量逆转录聚合酶链式反应、酶联免疫吸附试验、免疫印迹和流式细胞仪检测免疫细胞表型。流式细胞仪和免疫荧光染色检测miR-99b介导的巨噬细胞吞噬功能和抗原提呈能力。通过qRT-PCR、报告基因分析和蛋白质印迹分析，进一步验证了其潜在的分子机制。检测肝癌患者肿瘤组织及癌旁组织中miR-99b及其靶基因的表达。结果靶向转导miR-99b和/或miR-125a可显著抑制肝癌和LLC的生长，尤以miR-99b转导后更为明显。更重要的是，miR-99b的交付使重新接受了抗肿瘤表型的教育，增强了免疫监视。进一步的机制研究表明，巨噬细胞特异性过表达miR-99b分别通过靶向κB-RAS2和/或mTOR促进M1而抑制M2巨噬细胞极化。MIR-99b高表达的M1巨噬细胞具有较强的吞噬能力和抗原提呈能力。此外，将siMIR或siκB-RAS2导入TAMS可抑制miR-99b反义寡核苷酸引发的肿瘤生长。肝癌组织中miR-99b的表达低于癌旁组织，而κB-RAS2和mTOR的表达则相反。结论我们的结果揭示了miR-99b介导的表型的机制，表明靶向转导miR-99b是一种潜在的肿瘤免疫治疗策略。
Background Accumulating evidence has shown that tumor-associated macrophages (TAMs) play a critical role in tumor progression. Targeting TAMs is a potential strategy for tumor immunotherapy. However, the mechanism underlying the TAM phenotype and function needs to be resolved. Our previous studies have demonstrated that miR-125a can reverse the TAM phenotype toward antitumor. Meanwhile, we have found that miR-125a and miR-99b cluster in the first intron of the same host gene, and are transcribed simultaneously in bone marrow-derived macrophages (BMDMs) following LPS+IFNγ stimulation. However, it remains unclear whether miR-99b by itself can exert an antitumor effect by regulating macrophage phenotype. Methods miR-99b and/or miR-125a were delivered into TAMs of orthotopic hepatocellular carcinoma (HCC) or subcutaneous Lewis lung cancer (LLC) mice. The effect of treatment was evaluated by live imaging, TUNEL staining and survival tests. The phenotype of the immune cells was determined by qRT-PCR, ELISA, western blot and FACS. The capability of miR-99b-mediated macrophage phagocytosis and antigen presentation was detected by FACS and immunofluorescence staining. The underlying molecular mechanism was examined by qRT-PCR, reporter assay and western blot, and further verified in the tumor model. The expression of miR-99b and its target genes was determined in TAMs sorted from tumor and adjacent tissues in patients with liver cancer. Results Targeted delivery of miR-99b and/or miR-125a into TAMs significantly impeded the growth of HCC and LLC, especially after miR-99b delivery. More importantly, the delivery of miR-99b re-educated TAM toward antitumor phenotype with enhanced immune surveillance. Further investigation of mechanisms showed that macrophage-specific overexpression of miR-99b promoted M1 while suppressing M2 macrophage polarization by targeting κB-Ras2 and/or mTOR, respectively. miR-99b-overexpressed M1 macrophage was characterized by stronger capability of phagocytosis and antigen presentation. Additionally, delivery of simTOR or siκB-Ras2 into TAMs inhibited miR-99b antagomir-triggered tumor growth. Finally, miR-99b expression was lower in TAMs of patients with liver cancer than that in adjacent tissues, while the expression of κB-Ras2 and mTOR was reversed. Conclusions Our results reveal the mechanism of miR-99b-mediated TAM phenotype, indicating that TAM-targeted delivery of miR-99b is a potential strategy for cancer immunotherapy.
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影响因子：
3.7
作者：
Petty RD;McCarthy NE;Le Dieu R;Kerr JR
通讯作者：
Kerr JR
影响因子：
3.3
作者：
Saric A;Hipolito VE;Kay JG;Canton J;Antonescu CN;Botelho RJ
通讯作者：
Botelho RJ
影响因子：
29
作者：
Baker RG;Hayden MS;Ghosh S
通讯作者：
Ghosh S
miR-148a-3p 介导 notch 信号传导以促进巨噬细胞的分化和 M1 激活
影响因子：
7.3
作者：
Huang F;Zhao JL;Wang L;Gao CC;Liang SQ;An DJ;Bai J;Chen Y;Han H;Qin HY
通讯作者：
Qin HY
DOI：
10.1038/nrclinonc.2016.217
发表时间：
2017-07
期刊：
Nature reviews. Clinical oncology
影响因子：
--
作者：
Mantovani A;Marchesi F;Malesci A;Laghi L;Allavena P
通讯作者：
Allavena P