Zhang Beibei, Shimada Yasuhito, Hirota Tomokazu, Ariyoshi Michiko, Kuroyanagi Junya, Nishimura Yuhei, Tanaka Toshio
Department of Molecular and Cellular Pharmacology, Mie University Graduate School of Medicine, Mie, Japan; Department of Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Mie, Japan.
Department of Molecular and Cellular Pharmacology, Mie University Graduate School of Medicine, Mie, Japan; Department of Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Mie, Japan; Department of Systems Pharmacology, Mie University Graduate School of Medicine, Mie, Japan; Mie University Medical Zebrafish Research Center, Mie, Japan; Department of Bioinformatics, Mie University Life Science Research Center, Mie, Japan; Department of Omics Medicine, Mie University Industrial Technology Innovation Institute, Mie, Japan.
Transl Res. 2016 Apr;170:89-98.e3. doi: 10.1016/j.trsl.2015.12.007. Epub 2015 Dec 18.
Immune deficiency or suppression in host animals is an essential precondition for the success of cancer cell xenotransplantation because the host immune system has a tendency to reject implanted cells. However, in such animals, the typical tumor microenvironment seen in cancer subjects does not form because of the lack of normal immunity. Here, we developed a novel zebrafish (Danio rerio) model based on 2 rounds of cancer cell xenotransplantation that achieved cancer-specific immunologic tolerance without immunosuppression. We irradiated human cancer cells (PC-3, K562 and HepG2) to abolish their proliferative abilities and implanted them into zebrafish larvae. These cells survived for 2 weeks in the developing host. Three months after the first implantation, the zebrafish were implanted with the same, but nonirradiated, cell lines. These cancer cells proliferated and exhibited metastasis without immune suppression. To reveal the transcriptional mechanism of this immune tolerance, we conducted dual RNA-seq of the tumor with its surrounding tissues and identified several regulatory zebrafish genes that are involved in immunity; the expression of plasminogen activator, urokinase, and forkhead box P3 was altered in response to immunologic tolerance. In conclusion, this xenograft method has potential as a platform for zebrafish-based anticancer drug discovery because it can closely mimic human clinical cancers without inducing immune suppression.
宿主动物的免疫缺陷或抑制是癌细胞异种移植成功的必要前提，因为宿主免疫系统有排斥植入细胞的倾向。然而，在这类动物中，由于缺乏正常免疫力，癌症患者体内典型的肿瘤微环境无法形成。在此，我们基于两轮癌细胞异种移植开发了一种新型斑马鱼（Danio rerio）模型，该模型在不进行免疫抑制的情况下实现了癌症特异性免疫耐受。我们对人类癌细胞（PC-3、K562和HepG2）进行辐照以消除其增殖能力，然后将其植入斑马鱼幼体中。这些细胞在发育中的宿主体内存活了2周。首次植入三个月后，将相同但未辐照的细胞系植入斑马鱼体内。这些癌细胞在没有免疫抑制的情况下增殖并发生转移。为了揭示这种免疫耐受的转录机制，我们对肿瘤及其周围组织进行了双RNA测序，并鉴定了几个参与免疫的斑马鱼调控基因；纤溶酶原激活剂、尿激酶和叉头框P3的表达因免疫耐受而发生改变。总之，这种异种移植方法有潜力作为基于斑马鱼的抗癌药物发现平台，因为它可以在不诱导免疫抑制的情况下紧密模拟人类临床癌症。
