Joshi Bharat H, Suzuki Akiko, Fujisawa Toshio, Leland Pamela, Varrichio Frederick, Lababidi Samir, Lloyd Ricardo, Kasperbauer Jan, Puri Raj K
Tumor Vaccines and Biotechnology Branch, Division of Cellular and Gene Therapies, Office of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993, USA.
Office of Biostatistics and Epidemiology, Mayo Clinic, Minnesota, MN 55905, USA.
Discov Med. 2015 Nov;20(111):273-84.
Thyroid cancer is a rapidly increasing endocrine cancer. Since interleukin-4 receptor (IL-4R) is overexpressed in human solid cancer, we examined expression of IL-4R in 50 cases of anaplastic thyroid cancer (ATC), 37 well-differentiated papillary cancer (WDPC), 35 well-differentiated follicular cancer of thyroid (WDFC), and 37 normal thyroid specimens by immunohistochemistry (IHC) and in-situ hybridization (ISH) techniques. We demonstrated that IL-4Rα was overexpressed in 36/50 (72%) ATC, 20/35 (57%) WDFC, and 11/37 (30%) WDPC tumors. Other two subunits of IL-4R, interleukin-13 receptor α1 (IL-13Rα1) and interleukin-2 receptor gamma (IL-2RγC), were either weakly expressed or absent. As ATC is a highly aggressive cancer with higher incidence of IL-4Rα expression, we characterized IL-4R in 3 ATC cell lines. RT-qPCR and IFA results showed that IL-4Rα is overexpressed while IL-13Rα1 is weakly expressed. Control human umbilical vein endothelial cell line (HUVEC) showed weak expression of IL-4Rα. Binding and competition studies with 125I-IL-4 in ATC cell lines demonstrated that IL-4 specifically bound to IL-4Rα on cell surface. ATC cell lines were highly sensitive to a chimeric fusion cytotoxin consisting of circularly permuted IL-4 and truncated Pseudomonas exotoxin (IL-4-PE), which killed them in a concentration dependent manner. IL-4-PE also blocked colony formation of ATC cell lines in clonogenic assays. IL-4-PE mediated a significant antitumor activity in mouse models of ATC. Intratumoral administration of IL-4-PE caused significant regression of established tumors in a dose dependent manner and increased the overall survival without any visible toxicity. Thus, IL-4Rα in ATC may represent a novel therapeutic target and IL-4-PE may serve as an investigational therapeutic option for ATC.
甲状腺癌是一种发病率迅速上升的内分泌癌。由于白细胞介素4受体(IL-4R)在人类实体癌中过度表达,我们采用免疫组织化学(IHC)和原位杂交(ISH)技术检测了50例间变性甲状腺癌(ATC)、37例高分化乳头状癌(WDPC)、35例高分化甲状腺滤泡癌(WDFC)以及37例正常甲状腺标本中IL-4R的表达情况。我们发现,IL-4Rα在36/50(72%)的ATC、20/35(57%)的WDFC以及11/37(30%)的WDPC肿瘤中过度表达。IL-4R的其他两个亚基,即白细胞介素13受体α1(IL-13Rα1)和白细胞介素2受体γ(IL-2RγC),表达较弱或缺失。由于ATC是一种侵袭性很强的癌症,且IL-4Rα表达率较高,我们对3种ATC细胞系中的IL-4R进行了特性分析。逆转录定量聚合酶链反应(RT-qPCR)和免疫荧光分析(IFA)结果显示,IL-4Rα过度表达,而IL-13Rα1表达较弱。对照人脐静脉内皮细胞系(HUVEC)中IL-4Rα表达较弱。对ATC细胞系进行的125I-IL-4结合及竞争研究表明,IL-4能特异性结合细胞表面的IL-4Rα。ATC细胞系对由环化排列的IL-4和截短的绿脓杆菌外毒素组成的嵌合融合细胞毒素(IL-4-PE)高度敏感,该毒素能以浓度依赖的方式杀死细胞。在克隆形成试验中IL-4-PE还能阻断ATC细胞系的集落形成。在ATC小鼠模型中,IL-4-PE介导了显著的抗肿瘤活性。瘤内注射IL-4-PE能使已形成的肿瘤以剂量依赖的方式显著消退,并延长总生存期,且无任何明显毒性。因此,ATC中的IL-4Rα可能代表一种新的治疗靶点,而IL-4-PE可能作为ATC的一种试验性治疗选择。