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）在与癌症相关的死亡率中排名第三。 抑制抗肿瘤免疫学的调节T细胞（Tregs）的肿瘤内积累已被鉴定 在HCC中。尽管MicroRNA在HCC中进行了广泛的研究，但它们在确定免疫抑制中的作用是 描述不佳。 Akt的异常激活（V-Akt鼠胸腺瘤病毒癌同源1）和NRA （神经母细胞瘤RAS病毒癌基因同源物）（RAS）在Kupffer细胞（KCS）和肝细胞（HCS）中观察到（HCS） 在74％的HCC患者中。 Akt和NRA（AKT/RAS）活化形式的流体动力注射（HDI） 在6-8周内触发了致命HCC的发展。 miRNA分析表明miR-15a和miR-16- Akt/Ras小鼠和患者的HCC肿瘤的KC中减少了1个簇（miR-15a/16）。 KC特异性 miR-15a/16的表达完全防止了AKT/RAS小鼠的侵略性HCC的生长，而AKT/RAS的100％ 注射后6-8周内，小鼠死于致命的肿瘤燃烧。 MiR-15a/16的KC特异性表达 导致AKT/RAS小鼠的HCC肿瘤显着消退。机械上，AKT/RAS 将KC的转录组重新编程为KC的M2极化，并驱动了串行的显着增加 C-C基序趋化因子22（CCL22）和AKT/RAS小鼠KC中的CCL22的mRNA水平。相反，mir-15a/16 逆转了这一过程，并通过直接靶向激活转录的NF-κB来抑制CCL22过量生产 CCL22。 CCL22通过CCR4（C-C趋化因子受体4型）招募Treg。确实，CCL22的其他处理 在AKT/RAS小鼠中miR-15a/16完全预防的HCC的免疫抑制和生长。 我们假设AKT/RAS教育的KCS通过过量生产CCL22，从而启动Tregs的肝募集 从而促进HCC开发；而miR-15a/16的KC特异性表达抑制了肝募集 通过减弱CCL22生产来削弱Tregs和HCC开发。该项目的目的是阐明 AKT/RAS教育的KC在促进HCC生长的免疫抑制中的作用，而miR-15a/16 通过抑制KC中的NF-κB信号传导来逆转此过程。长期目标是开发KC作为一种疗法。 靶标和miR-15a/16作为针对HCC的新型免疫疗法。三个特定目标旨在测试我们 假设。具体而言，我们将（1）阐明AKT/RAS教育的KC促进的机制 免疫抑制和HCC开发。 （2）建立mir-15a/16驱动器的机制 抗肿瘤免疫； （3）评估miR-15a/16对HCC的治疗潜力 背景。强大的初步数据和基于逻辑和合理的实验设计的组合 使这个项目非常可行。我们希望这项研究将为免疫抑制提供新的见解 HCC中的机制，并促进KCS作为一种新型的治疗靶标，而MiR-15A/16作为理性 针对HCC的免疫疗法。