Establishment of an experimental model of normal dog bladder organoid using a three-dimensional culture method
Establishment of an experimental model of normal dog bladder organoid using a three-dimensional culture method
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DOI：
10.1016/j.biopha.2022.113105
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发表时间：
2022-05-20
影响因子：
7.5
通讯作者：
Sasaki, Kazuaki
中科院分区：
文献类型：
--
作者：
Elbadawy, Mohamed;Fujisaka, Kodai;Sasaki, Kazuaki
犬膀胱癌（BC）大多为肌层浸润性（MI），预后不良，其发病机制与人类肌层浸润性膀胱癌（MIBC）相近。三维（3D）类器官培养为包括膀胱癌在内的癌症疾病研究提供了新的知识。近期，我们利用犬的尿液样本建立了犬膀胱癌类器官（BCO）。BCO重现了患膀胱癌犬的上皮结构、特征以及药物敏感性。然而，犬正常膀胱上皮的类器官尚未建立。因此，本研究旨在建立犬正常膀胱类器官（NBO），以进一步了解犬膀胱癌和人类MIBC的发病机制。所建立的NBO进行了多种分析，包括细胞标志物表达、组织病理结构、癌症相关基因表达模式以及药物敏感性。NBO能够通过无创方式培养且可连续培养，并重现了犬正常膀胱黏膜组织的结构和特征。在每个NBO中观察到了不同的药物敏感性。RNA测序分析显示，与BCO相比，NBO中有几个新基因发生了变化。与BCO相比，NBO中p53和E - 钙黏蛋白表达较高，而MDM2和Twist1表达较低。这些结果表明，NBO可能是一个有前景的用于研究犬膀胱癌和人类MIBC发生机制的实验性3D模型。
Dog bladder cancer (BC) is mostly muscle-invasive (MI) with poor prognosis, and its pathogenesis is close to human MIBC. Three-dimensional (3D) organoid culture ensures novel knowledge on cancer diseases including BC. Recently, we have established dog BC organoids (BCO) using their urine samples. BCO recapitulated the epithelial structures, characteristics, and drug sensitivity of BC-diseased dogs. However, organoids from dog normal bladder epithelium are not established yet. Therefore, the present study aimed to establish dog normal bladder organoids (NBO) for further understanding the pathogenesis of dog BC and human MIBC. The established NBO underwent various analyzes including cell marker expressions, histopathological structures, cancer-related gene expression patterns, and drug sensitivity. NBO could be produced non-invasively with a continuous culturing and recapitulated the structures and characteristics of the dog's normal bladder mucosal tissues. Different drug sensitivities were observed in each NBO. The analysis of RNA sequencing revealed that several novel genes were changed in NBO compared with BCO. NBO showed a higher expression of p53 and E-cadherin, but a lower expression of MDM2 and Twist1 compared with BCO. These results suggest that NBO could be a promising experimental 3D model for studying the developmental mechanisms of dog BC and human MIBC.