Molecular analysis of dendritic cell based vaccines against liver cancer

基于树突状细胞的肝癌疫苗的分子分析

• 批准号: 17590638
• 负责人: TATSUMI Tomohide
• 金额: $ 2.24万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590638/
• 关键词: Liver cancer; Dendritic cell; Cancer Immunotherapy; Interleukin 12; EphA2; α-galactosylceramide

(1) Efficacy of immunothetrapy with the receptor tyrosine kinase EphA2 derived peptides-pulsed dendritic cells in mouse liver tumor.The receptor tyrosine kinase EphA2,a new tumor-associated antigen, is broadly expressed in diverse cancer, especially in advanced-stage cancer or metastatic cancer. In this study, we evaluated the effectiveness of the immunization with EphA2 derived peptide-pulsed DCs in mouse subcutaneous and liver tumor. IFN-Υ ELISPOT assays revealed that EphA2 derived peptides-specific CTLs could be generated in vivo by immunization of EphA2 peptide pulsed DCs (Eph-DCs). MC38 subcutaneous or liver tumor-bearing mice were treated with Eph-DCs or unpulsed DCs. MC38 subcutaneous tumor growth in the mice treated with Eph-DCs was significantly inhibited compared with that in the-mice treated with unpulsed DCs. In contrast, MC38 liver tumor growth was significantly inhibited in the mice treated with both Eph-DCs or unpulsed DCs. In vivo depletion study revealed that CD8+ T ce … More lls mainly, not NK cells, played critical roles for antitumor effect of Eph-DCs treatment in subcutaneous tumor. In liver tumor model, not only CD8+ T cells but also NK cells played critical roles in antitumor effect of Eph-DCs. These results demonstrated that immunization of Eph-DCs activated acquired immunity in subcutaneous tissue and both innate and acquired immunity in the liver, suggesting the therapeutic potential of Eph-DCs vaccine against EphA2-expressing subcutaneous and liver tumor.(2) Injection of IL-12 gene-transduced dendritic cells into mouse liver tumor lesions activates both innate and acquired immunityDendritic cells (DCs) based vaccines have been applied clinically in the setting of advanced-stage cancer. To date, the clinical efficacy of these vaccines has been limited, possibly due to the impairment of transferred DC function in cancer bearing-patients. In this study, we examined the therapeutic efficacy of IL-12 gene-transfected DCs isolated from tumor bearing hosts against liver tumor. The endogenous DCs isolated from subcutaneous CMS4 tumor-bearing mice (CMS4DC) exhibited decreased DC function compared with DCs isolated from normal mice. Adenoviral transfection of IL-12 gene into CMS4DC (AdIL12DC) restored the decreased DC function. Intratumoral (i.t.) delivery of AdIL12DC resulted in complete rejection of intrahepatic CMS4 tumors and activation of innate and acquired immune cells. I.t. treatment of AdIL12DC also resulted in long-term protection against s.c. rechallenge with CMS4 tumor cells. These results revealed that IL-12 gene transfer is capable of improving the impaired functions of DC isolated from tumor bearing hosts, and support the preclinical therapeutic efficacy of intrahepatic injection of AdIL12DC.(3) Intrahepatic delivery of α-Galactosylceramide pulsed dendritic cells (αGCDC) suppresses liver tumor.α-Galactosylceramide mediates interaction of DCs and NKT cells, leading to activation of both innate and acquired immunity. For cancer treatment, conventional DC-based vaccine has been tried, but its clinical efficacy is limited against liver cancer. Intrahepatic injection of αGCDC has not yet been tested in the liver that contains abundant immune cells such as NK, NKT and T cells. We examined the efficacy of αGCDC administration in comparison with p53 peptide-pulsed DCs using a well-established murine CMS4 tumor model. Injection of αGCDC into CMS4 liver tumors resulted in complete tumor rejection and established long-term survival of the animals, while injection of p53_<232-240>o peptide-pulsed DCs (pepDC) only partially suppressed tumor growth in the liver. Hepatic NK cells were efficiently activated by αGCDC injection and played a critical role in liver tumor rejection as evidenced by an in vivo antibody-mediated NK cell depletion study. Injection of αGCDC into liver tumor led to higher p53_<232-240> peptide-specific CD8+ T cell response than that of pepDC. The mice that had been protected from CMS4 liver tumor by αGCDC injection became resistant to subcutaneous CMS4 rechallenge, but not to Colon26 rechallenge. These results demonstrate that αGCDC injection into the liver can efficiently activate NK cells that in turn reject liver tumors to establish potent acquired immunity against the original tumor. Less

(1)受体酪氨酸激酶EphA2衍生肽脉冲树突状细胞免疫治疗小鼠肝肿瘤的疗效。受体酪氨酸激酶EphA2是一种新的肿瘤相关抗原，在多种癌症中广泛表达，特别是在晚期癌症或转移性癌症中。在这项研究中，我们评估了EphA2衍生肽脉冲dc对小鼠皮下和肝脏肿瘤的免疫效果。IFN-Υ ELISPOT检测结果显示，EphA2肽脉冲dc （EphA2 peptide pulsed DCs）可在体内产生EphA2衍生肽特异性ctl。用epf - dc或无脉冲dc治疗MC38皮下或肝脏荷瘤小鼠。与未脉冲dc处理的小鼠相比，ef - dc处理的小鼠MC38皮下肿瘤生长明显受到抑制。相比之下，用epf - dc或未脉冲dc处理的小鼠MC38肝肿瘤生长均受到显著抑制。体内耗竭实验表明，epf - dc对皮下肿瘤的抗肿瘤作用主要是CD8+ T细胞，而不是NK细胞。在肝脏肿瘤模型中，除了CD8+ T细胞外，NK细胞也参与了epf - dc的抗肿瘤作用。这些结果表明，免疫Eph-DCs可激活皮下组织获得性免疫和肝脏先天免疫和获得性免疫，提示Eph-DCs疫苗对表达epha2的皮下和肝脏肿瘤具有治疗潜力。(2)将IL-12基因转导的树突状细胞注射到小鼠肝脏肿瘤病变处，可激活先天免疫和获得性免疫。基于树突状细胞（dc）的疫苗已在临床上用于晚期癌症的治疗。迄今为止，这些疫苗的临床疗效有限，可能是由于癌症患者转移的DC功能受损。在本研究中，我们检测了从荷瘤宿主分离的转染IL-12基因的dc对肝脏肿瘤的治疗效果。从皮下CMS4荷瘤小鼠（CMS4DC）中分离的内源性DC与正常小鼠分离的DC相比，DC功能下降。腺病毒转染IL-12基因到CMS4DC （AdIL12DC）后，恢复了下降的DC功能。肿瘤内（i.t）递送AdIL12DC导致肝内CMS4肿瘤的完全排斥和先天和获得性免疫细胞的激活。AdIL12DC的治疗也对CMS4肿瘤细胞的sc再攻击产生长期保护作用。这些结果表明IL-12基因转移能够改善荷瘤宿主DC的功能受损，支持肝内注射AdIL12DC的临床前治疗效果。(3) α-半乳糖神经酰胺脉冲树突状细胞（α - gcdc）肝内递送抑制肝脏肿瘤。α-半乳糖神经酰胺介导dc和NKT细胞的相互作用，导致先天免疫和获得性免疫的激活。对于癌症的治疗，传统的基于dc的疫苗已经尝试过，但其对肝癌的临床疗效有限。肝内注射α - gcdc尚未在含有大量NK、NKT和T细胞等免疫细胞的肝脏中进行试验。我们通过建立小鼠CMS4肿瘤模型，比较α - gcdc给药与p53肽脉冲dc的疗效。在CMS4肝肿瘤中注射α - gcdc可引起肿瘤的完全排斥反应并建立动物的长期生存，而注射p53_<232-240>肽脉冲dc （pepDC）仅能部分抑制肝脏肿瘤的生长。一项体内抗体介导的NK细胞耗竭研究证实，α - gcdc可有效激活肝脏NK细胞，并在肝脏肿瘤排斥反应中发挥关键作用。肝肿瘤注射α - gcdc后，p53_<232 ~ 240>肽特异性CD8+ T细胞反应高于pepDC。经α - gcdc保护的小鼠对CMS4肝肿瘤皮下再攻毒产生抗性，但对Colon26无抗性。上述结果表明，α - gcdc注射肝脏可有效激活NK细胞，NK细胞对肝脏肿瘤产生排斥反应，从而建立对原肿瘤的获得性免疫。少

项目成果

Impairment of natural killer cell and dendritic cell functions by the soluble form of MHC class I-related chain A in advanced human hepatocellular carcinoma.

MHC I 类相关链 A 在晚期人肝细胞癌中可溶形式对自然杀伤细胞和树突状细胞功能的损害。

• 发表时间: 2005
• 期刊: Journal of Hepatology 43
• 作者: Jinushi M.;Tatsumi T.;et al.
• 通讯作者: et al.

Expression of EphA2 is prognostic of disease-free interval and overall survivals in surgically treated patients with renal cell carcinoma

EphA2 的表达可预测手术治疗的肾细胞癌患者的无病间隔和总生存率

• 发表时间: 2005
• 期刊: Clinical Cancer Research 11
• 作者: Herrem C.J.;Tatsumi T.;et al.
• 通讯作者: et al.

Injection of IL-12 gene-transduced dendritic cells into mouse liver tumor lesions activates both innate and acquired immunity

• DOI: 10.1038/sj.gt.3302941
• 发表时间: 2007-06
• 期刊: Gene Therapy
• 影响因子: 5.1
• 作者: T. Tatsumi;Tetsuo Takehara;Shinjiro Yamaguchi;Akira Sasakawa;T. Miyagi;M. Jinushi;R. Sakamori;Keisuke Kohga;A. Uemura;K. Ohkawa;W. Storkus;Norio Hayashi

Intrahepatic delivery ofα-galatosylceramide pulsed dendriticcells suppresses liver tumor

α-半乳糖神经酰胺脉冲树突状细胞的肝内递送抑制肝脏肿瘤

• 发表时间: 2007
• 期刊: Hepatology 45
• 作者: Tatsumi T(1番目);Takehara T(2番目);(他7名)
• 通讯作者: (他7名)

Nitric Oxide sensitizes tumor cells to dendritic cells-mediated apoptosis, uptake and cross-presentation

一氧化氮使肿瘤细胞对树突状细胞介导的细胞凋亡、摄取和交叉呈递敏感

• 发表时间: 2005
• 期刊: Cancer Research 65
• 作者: Huang J.;Tatsumi T.;et al.
• 通讯作者: et al.