Genetically engineered rat gliomas: PDGF-driven tumor initiation and progression in tv-a transgenic rats recreate key features of human brain cancer.
Genetically engineered rat gliomas: PDGF-driven tumor initiation and progression in tv-a transgenic rats recreate key features of human brain cancer.
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DOI：
10.1371/journal.pone.0174557
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Woodworth GF
中科院分区：
文献类型：
--
作者：
Connolly NP;Stokum JA;Schneider CS;Ozawa T;Xu S;Galisteo R;Castellani RJ;Kim AJ;Simard JM;Winkles JA;Holland EC;Woodworth GF
先前的啮齿动物胶质瘤临床前研究通常涉及将细胞系如C6和9L植入大鼠大脑。最近，小鼠模型已经占据了主导地位，小鼠的遗传可操控性允许创建基因精确的模型，这超过了其较小的大脑尺寸的缺点，即有限的时间允许肿瘤进展。在这里，我们说明了一种方法，允许在大鼠胶质瘤形成使用复制能力的鸟样肉瘤（RCAS）病毒/肿瘤病毒受体-a （tv-a）转基因系统的出生后细胞类型特异性基因转移。RCAS/tv-a模型已经成为一种特别通用和精确的建模技术，通过实现对单个基因转化的空间、时间和细胞类型特异性控制，并提供具有不同分子亚型的新生胶质肿瘤，反映了人类GBM。构建巢蛋白启动子驱动的tv-a （Ntv-a）转基因Sprague-Dawley大鼠方正系，并使用RCAS PDGFA和p53 shRNA构建物启动颅内脑肿瘤形成。通过磁共振成像（MRI）和光谱学证实和观察肿瘤的形成和进展。采用组织病理学和免疫荧光技术对肿瘤进行分析。所有的实验动物都长出了巨大的异质脑瘤，与人类GBM非常相似。肿瘤发生后的中位生存期为92天，MRI显示肿瘤后的中位生存期为62天。通过MRI和神经学检查，每只荷瘤动物都表现出恶性进展为高级别胶质瘤的时间依赖性证据。死后肿瘤分析显示了人类GBM的几个关键特征，包括高水平的肿瘤细胞增殖、假乳样坏死、微血管增殖、肿瘤细胞侵入周围组织、肿瘤周围反应性星形胶质细胞形成、淋巴细胞浸润、大量肿瘤相关小胶质细胞和骨髓源性巨噬细胞的存在，以及肿瘤内干细胞样细胞龛的形成。这种转基因大鼠模型可以对基本的癌症途径和临床相关的实验成像程序和干预进行详细的物种间比较，这些都受到小鼠大脑较小尺寸的限制。
Previously rodent preclinical research in gliomas frequently involved implantation of cell lines such as C6 and 9L into the rat brain. More recently, mouse models have taken over, the genetic manipulability of the mouse allowing the creation of genetically accurate models outweighed the disadvantage of its smaller brain size that limited time allowed for tumor progression. Here we illustrate a method that allows glioma formation in the rat using the replication competent avian-like sarcoma (RCAS) virus / tumor virus receptor-A (tv-a) transgenic system of post-natal cell type-specific gene transfer. The RCAS/tv-a model has emerged as a particularly versatile and accurate modeling technology by enabling spatial, temporal, and cell type-specific control of individual gene transformations and providing de novo formed glial tumors with distinct molecular subtypes mirroring human GBM. Nestin promoter-driven tv-a (Ntv-a) transgenic Sprague-Dawley rat founder lines were created and RCAS PDGFA and p53 shRNA constructs were used to initiate intracranial brain tumor formation. Tumor formation and progression were confirmed and visualized by magnetic resonance imaging (MRI) and spectroscopy. The tumors were analyzed using histopathological and immunofluorescent techniques. All experimental animals developed large, heterogeneous brain tumors that closely resembled human GBM. Median survival was 92 days from tumor initiation and 62 days from the first point of tumor visualization on MRI. Each tumor-bearing animal showed time dependent evidence of malignant progression to high-grade glioma by MRI and neurological examination. Post-mortem tumor analysis demonstrated the presence of several key characteristics of human GBM, including high levels of tumor cell proliferation, pseudopalisading necrosis, microvascular proliferation, invasion of tumor cells into surrounding tissues, peri-tumoral reactive astrogliosis, lymphocyte infiltration, presence of numerous tumor-associated microglia- and bone marrow-derived macrophages, and the formation of stem-like cell niches within the tumor. This transgenic rat model may enable detailed interspecies comparisons of fundamental cancer pathways and clinically relevant experimental imaging procedures and interventions that are limited by the smaller size of the mouse brain.