MicroRNA-15a/16-mediated cytokine/chemokine reprogramming in Kupffer cells prevents the development of hepatocellular carcinoma

MicroRNA-15a/16 介导的库普弗细胞细胞因子/趋化因子重编程可预防肝细胞癌的发展

• 批准号: 10518314
• 负责人: Guisheng Song
• 金额: $ 37.25万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-22 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10518314
• 关键词: Attenuated; Automobile Driving; Blood; CCL22 gene; CRISPR/Cas technology; Cell Therapy; Cells; Country; Cytotoxic T-Lymphocytes; Data; Development; Diethylnitrosamine; Etiology; Experimental Designs; Genetic Transcription; Goals; Grant; Growth; Hepatic; Hepatocyte; Homologous Gene; Immune; Immunosuppression; Immunotherapy; Impairment; In Vitro; Incidence; Injections; Kupffer Cells; Lead; Liver; Liver neoplasms; Malignant Neoplasms; Mediating; Messenger RNA; MicroRNAs; Mus; NRAS gene; Obesity; Oncogenes; Oncogenic; Patients; Primary carcinoma of the liver cells; Process; Production; Proto-Oncogene Proteins c-akt; Regulatory T-Lymphocyte; Research; Role; Serum; Signal Transduction; T-Cell Depletion; Techniques; Testing; Therapeutic; Thymoma; Time; Transplantation; Tumor Burden; Tumor Immunity; Viral Oncogene; base; beta-Chemokines; c-myc Genes; chemokine; chemokine receptor; cytokine; design; in vivo Model; ineffective therapies; insight; knock-down; liver cancer model; migration; mortality; mouse model; new therapeutic target; novel; prevent; recruit; success; therapeutic miRNA; therapeutic target; transcriptome; tumor

Abstract Hepatocellular carcinoma (HCC) ranks the third in cancer-related mortality because of ineffective therapy. Intratumoral accumulation of regulatory T-cells (Tregs), which suppresses antitumor immunity, has been identified in HCC. Despite the extensive research of microRNAs in HCC, their roles in regulating immunosuppression are poorly described. Aberrant activation of AKT (v-akt murine thymoma viral oncogene homolog 1) and Nras (neuroblastoma ras viral oncogene homolog) (Ras) was observed in Kupffer cells (KCs) and hepatocytes (HCs) in 74% of HCC patients. Hydrodynamic injection (HDI) of activated forms of AKT and Nras (AKT/Ras) into mice triggered the development of lethal HCC within 6-8 weeks. MiRNA profiling revealed that miR-15a and miR-16- 1 cluster (miR-15a/16) was reduced in KCs from HCC tumors of AKT/Ras mice and patients. KC-specific expression of miR-15a/16 fully prevented growth of aggressive HCC in AKT/Ras mice, while 100% of AKT/Ras mice died from lethal tumor burden within 6-8 weeks post-injection. KC-specific expression of miR-15a/16 also led to a significant regression of HCC tumors in AKT/Ras mice bearing tumors. Mechanistically, AKT/Ras reprogrammed the transcriptome of KCs toward M2 polarization of KCs and drove a significant increase in serum C-C motif chemokine 22 (CCL22) and mRNA levels of Ccl22 in KCs of AKT/Ras mice. In contrast, miR-15a/16 reversed this process and inhibited CCL22 overproduction by directly targeting NF-κB that activates transcription of Ccl22. CCL22 recruits Tregs via CCR4 (C-C chemokine receptor type 4). Indeed, additional treatment of CCL22 recovered immunosuppression and growth of HCC that was fully prevented by miR-15a/16 in AKT/Ras mice. We hypothesize that AKT/Ras-educated KCs initiate hepatic recruitment of Tregs by overproducing CCL22, thereby promoting HCC development; while KC-specific expression of miR-15a/16 suppresses hepatic recruitment of Tregs and HCC development by attenuating CCL22 production. The objective of this project is to elucidate the role of AKT/Ras-educated KCs in promoting immune suppression that promotes growth of HCC, while miR-15a/16 reverses this process by inhibiting NF-κB signaling in KCs. The long-time goal is to develop KCs as a therapeutic target and miR-15a/16 as a novel immunotherapy against HCC. Three specific aims are designed to test our hypothesis. Specifically, we will (1) unravel the mechanism(s) by which AKT/Ras-educated KCs promote immunosuppression and HCC development. (2) establish the mechanism(s) by which miR-15a/16 drives antitumor immunity; and (3) evaluate the therapeutic potential of miR-15a/16 against HCC with divergent backgrounds. The combination of strong preliminary data and a logical and rationally based experimental design make this project highly feasible. We expect that this study will provide a novel insight into the immunosuppression mechanisms in HCC and facilitate the design of KCs as a novel therapeutic target and miR-15a/16 as a rational immunotherapy against HCC.

抽象的 由于治疗无效，肝细胞癌（HCC）在癌症相关死亡率中排名第三。 已发现调节性 T 细胞 (Treg) 在肿瘤内积聚，可抑制抗肿瘤免疫 在肝癌中。尽管对 HCC 中的 microRNA 进行了广泛的研究，但它们在调节免疫抑制方面的作用仍不清楚。 描述得很差。 AKT（v-akt 鼠胸腺瘤病毒癌基因同源物 1）和 Nras 的异常激活 在库普弗细胞 (KC) 和肝细胞 (HC) 中观察到（神经母细胞瘤 ras 病毒癌基因同源物）(Ras) 74% 的 HCC 患者。将活化形式的 AKT 和 Nras (AKT/Ras) 流体动力注射 (HDI) 至小鼠体内 在 6-8 周内引发致命性 HCC 的发展。 miRNA 分析显示 miR-15a 和 miR-16- AKT/Ras 小鼠和患者 HCC 肿瘤的 KC 中 1 个簇 (miR-15a/16) 减少。 KC 特定 miR-15a/16 的表达完全阻止 AKT/Ras 小鼠中侵袭性 HCC 的生长，而 100% AKT/Ras 小鼠在注射后 6-8 周内死于致命的肿瘤负荷。 miR-15a/16 的 KC 特异性表达 导致荷瘤 AKT/Ras 小鼠的 HCC 肿瘤显着消退。从机制上讲，AKT/Ras 将 KC 的转录组重新编程为 KC 的 M2 极化，并导致血清显着增加 AKT/Ras 小鼠 KC 中的 C-C 基序趋化因子 22 (CCL22) 和 Ccl22 的 mRNA 水平。相比之下，miR-15a/16 通过直接靶向激活转录的 NF-κB，逆转了这一过程并抑制了 CCL22 的过量产生 Ccl22。 CCL22 通过 CCR4（C-C 趋化因子受体 4 型）招募 Tregs。事实上，CCL22 的额外治疗 在 AKT/Ras 小鼠中，miR-15a/16 完全阻止了免疫抑制和 HCC 生长的恢复。 我们假设 AKT/Ras 训练的 KC 通过过量产生 CCL22 来启动肝脏募集 Tregs， 从而促进HCC的发展；而 miR-15a/16 的 KC 特异性表达抑制肝脏募集 通过减少 CCL22 的产生来抑制 Tregs 和 HCC 的发展。该项目的目标是阐明 AKT/Ras 教育的 KC 在促进免疫抑制中的作用，从而促进 HCC 的生长，而 miR-15a/16 通过抑制 KC 中的 NF-κB 信号传导来逆转这一过程。长期目标是将 KC 开发为一种治疗方法 靶点和 miR-15a/16 作为针对 HCC 的新型免疫疗法。设计了三个具体目标来测试我们的 假设。具体来说，我们将 (1) 揭示受过 AKT/Ras 教育的 KC 促进的机制 免疫抑制和 HCC 的发展。 (2)建立miR-15a/16驱动的机制 抗肿瘤免疫力； (3) 评估 miR-15a/16 对 HCC 的治疗潜力 背景。强有力的初步数据与逻辑合理的实验设计相结合 使这个项目具有高度的可行性。我们期望这项研究将为免疫抑制提供新的见解 HCC 的机制并促进 KC 的设计作为新的治疗靶点和 miR-15a/16 作为合理的治疗靶点 针对 HCC 的免疫疗法。