Characterization of two rat models of cystic fibrosis—KO and F508del CFTR—Generated by Crispr-Cas9

Animal Models and Experimental Medicine

article

Elise Dreano¹  Marc Bacchetta²  Juliette Simonin²  Louise Galmiche³  Claire Usal⁴  Lotfi Slimani⁶  Jérémy Sadoine⁶  Laurent Tesson⁴  Ignacio Anegon⁴  Jean-Paul Concordet⁷  Aurélie Hatton¹  Lucile Vignaud¹  Danielle Tondelier¹  Isabelle Sermet-Gaudelus¹  Marc Chanson²  Charles-Henry Cottart¹ 

[1] INSERM 1151, INEM, Université de Paris;Département de Pédiatrie, Gynécologie & Obstétrique et Département de Physiologie Cellulaire & Métabolisme, Université de Genève;Département de Pathologie, APHP, CHU Necker-Enfants Malades;Centre de Recherche en Transplantation & Immunologie, Université de Nantes;Plateforme Trangénèse Rat & ImmunoPhénomique, INSERM 1064 & SFR François Bonamy;Pathologie, Imagerie & Biothérapies Orofaciales;INSERM U1154, CNRS UMR7196, MNHN;Centre Maladie Rare Mucoviscidose et Maladies du CFTR;Faculté de Médecine de Paris, Université de Paris;AP-HP, Centre Maladie Rare Mucoviscidose et Maladies du CFTR, Assistance Publique Hôpitaux de Paris, Hôpital Necker-Enfants Malades;Faculté de Pharmacie de Paris, Université de Paris

关键词: animal models;    CFTR channel activity;    CFTR modulators;    cystic fibrosis;    primary cultures;    rat;   

DOI  :  10.1002/ame2.12091

学科分类：机械工程学

来源: Wiley

【 摘 要 】

Background: 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. Methods: We describe the generation of a CF rat model (F508del) homozygous for the p.Phe508del mutation in the transmembrane conductance regulator (Cftr) gene. This model was compared to new Cftr−/− 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.Results: 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 vas deferens 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. Conclusions: The F508del rats reproduce the phenotypes observed in CFTR KO animals and represent a novel resource to advance the development of CF therapeutics.

【 授权许可】

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