Dreano Elise, Bacchetta Marc, Simonin Juliette, Galmiche Louise, Usal Claire, Slimani Lotfi, Sadoine Jérémy, Tesson Laurent, Anegon Ignacio, Concordet Jean-Paul, Hatton Aurélie, Vignaud Lucile, Tondelier Danielle, Sermet-Gaudelus Isabelle, Chanson Marc, Cottart Charles-Henry
INSERM 1151 INEM Université de Paris Paris France.
Département de Pédiatrie Gynécologie & Obstétrique et Département de Physiologie Cellulaire & Métabolisme Université de Genève Genève Switzerland.
Animal Model Exp Med. 2019 Nov 25;2(4):297-311. doi: 10.1002/ame2.12091. eCollection 2019 Dec.
Genetically engineered animals are essential for gaining a proper understanding of the disease mechanisms of cystic fibrosis (CF). The rat is a relevant laboratory model for CF because of its zootechnical capacity, size, and airway characteristics, including the presence of submucosal glands.
We describe the generation of a CF rat model (F508del) homozygous for the p.Phe508del mutation in the transmembrane conductance regulator () gene. This model was compared to new rats (CFTR KO). Target organs in CF were examined by histological staining of tissue sections and tooth enamel was quantified by micro-computed tomography. The activity of CFTR was evaluated by nasal potential difference (NPD) and short-circuit current measurements. The effect of VX-809 and VX-770 was analyzed on nasal epithelial primary cell cultures from F508del rats.
Both newborn F508del and Knock out (KO) animals developed intestinal obstruction that could be partly compensated by special diet combined with an osmotic laxative. The two rat models exhibited CF phenotypic anomalies such as agenesis and tooth enamel defects. Histology of the intestine, pancreas, liver, and lungs was normal. Absence of CFTR function in KO rats was confirmed ex vivo by short-circuit current measurements on colon mucosae and in vivo by NPD, whereas residual CFTR activity was observed in F508del rats. Exposure of F508del CFTR nasal primary cultures to a combination of VX-809 and VX-770 improved CFTR-mediated Cl transport.
The F508del rats reproduce the phenotypes observed in CFTR KO animals and represent a novel resource to advance the development of CF therapeutics.
基因工程动物对于正确理解囊性纤维化（CF）的疾病机制至关重要。大鼠因其畜牧学能力、体型以及气道特征（包括存在黏膜下腺），是一种与CF相关的实验模型。
我们描述了一种跨膜传导调节因子（CFTR）基因中p.Phe508del突变纯合的CF大鼠模型（F508del）的构建。将该模型与新的CFTR基因敲除（CFTR KO）大鼠进行比较。通过组织切片的组织学染色检查CF中的靶器官，并通过微计算机断层扫描对牙釉质进行定量分析。通过鼻电位差（NPD）和短路电流测量评估CFTR的活性。分析了VX - 809和VX - 770对F508del大鼠鼻上皮原代细胞培养物的影响。
新生的F508del和基因敲除（KO）动物均出现肠梗阻，特殊饮食联合渗透性泻药可部分缓解。这两种大鼠模型均表现出CF表型异常，如唾液腺发育不全和牙釉质缺陷。肠、胰腺、肝脏和肺的组织学检查正常。通过结肠黏膜的短路电流测量在体外证实了KO大鼠中CFTR功能的缺失，通过NPD在体内证实了这一点，而在F508del大鼠中观察到了残余的CFTR活性。将F508del CFTR鼻原代培养物暴露于VX - 809和VX - 770的组合中可改善CFTR介导的氯离子转运。
F508del大鼠再现了CFTR KO动物中观察到的表型，是推进CF治疗药物开发的一种新资源。
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