Teague Warwick J, Jones Matthew L M, Hawkey Leanne, Smyth Ian M, Catubig Angelique, King Sebastian K, Sarila Gulcan, Li Ruili, Hutson John M
F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia.
Department of Paediatrics, The University of Melbourne, Melbourne, VIC, Australia.
Front Genet. 2018 Nov 9;9:530. doi: 10.3389/fgene.2018.00530. eCollection 2018.
Duodenal atresia (DA) is a congenital obstruction of the duodenum, which affects 1 in 7000 pregnancies and requires major surgery in the 1st days of life. Three morphological DA types are described. In humans, the association between DA and Down syndrome suggests an underlying, albeit elusive, genetic etiology. In mice, interruption of fibroblast growth factor 10 () gene signaling results in DA in 30-50% of embryos, supporting a genetic etiology. This study aims to validate the spectrum of DA in two novel strains of knock-out mice, in preparation for future and translational research. Two novel CRISPR knock-out mouse strains were derived and embryos generated by heterozygous plug-mating. E15.5-E19.5 embryos were genotyped with respect to and micro-dissected to determine the presence and type of DA. One twenty seven embryos (32 wild-type, 34 heterozygous, 61 null) were analyzed. No wild-type or heterozygous embryos had DA. However, 74% of null embryos had DA (49% type 1, 18% type 2, and 33% type 3). Our CRISPR-derived strains showed higher penetrance of DA due to single-gene deletion of in mice than previously reported. Further, the DA type distribution in these mice more closely reiterated that observed in humans. Future experiments will document RNA and protein expression of FGF10 and its key downstream signaling targets in normal and atretic duodenum. This includes exploitation of modern, high-fidelity developmental tools, e.g., -tomato mice.
十二指肠闭锁（DA）是一种十二指肠先天性梗阻疾病，在每7000例妊娠中会出现1例，患病婴儿在出生后的头几天就需要进行大手术。DA有三种形态学类型。在人类中，DA与唐氏综合征之间的关联表明其存在潜在的遗传病因，尽管这种病因难以捉摸。在小鼠中，成纤维细胞生长因子10（FGF10）基因信号中断会导致30%-50%的胚胎出现DA，这支持了遗传病因的观点。本研究旨在验证两种新型FGF10基因敲除小鼠品系中DA的谱系，为未来的基础研究和转化研究做准备。我们获得了两种新型的CRISPR FGF10基因敲除小鼠品系，并通过杂合子交配产生胚胎。对E15.5-E19.5期胚胎进行FGF10基因分型，并进行显微解剖以确定DA的存在和类型。共分析了127个胚胎（32个野生型、34个杂合子、61个纯合缺失型）。没有野生型或杂合子胚胎出现DA。然而，74%的纯合缺失型胚胎出现了DA（49%为1型，18%为2型，33%为3型）。我们通过CRISPR技术获得的品系显示，由于小鼠中FGF10单基因缺失，DA的发生率比之前报道的更高。此外，这些小鼠中DA类型的分布更接近人类中观察到的情况。未来的实验将记录正常和闭锁十二指肠中FGF10及其关键下游信号靶点的RNA和蛋白质表达情况。这包括利用现代的、高保真的发育工具，例如Rosa26-tdTomato小鼠。
