Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, Ontario K1H 8L6, Canada.
Departments of Biochemistry, Microbiology, & Immunology, Ottawa, Ontario K1H 8M5, Canada.
Hum Mol Genet. 2021 May 12;30(7):575-594. doi: 10.1093/hmg/ddab081.
The PHF6 mutation c.1024C > T; p.R342X, is a recurrent cause of Börjeson-Forssman-Lehmann Syndrome (BFLS), a neurodevelopmental disorder characterized by moderate-severe intellectual disability, truncal obesity, gynecomastia, hypogonadism, long tapering fingers and large ears (MIM#301900). Here, we generated transgenic mice with the identical substitution (R342X mice) using CRISPR technology. We show that the p.R342X mutation causes a reduction in PHF6 protein levels, in both human and mice, from nonsense-mediated decay and nonsense-associated alternative splicing, respectively. Magnetic resonance imaging studies indicated that R342X mice had a reduced brain volume on a mixed genetic background but developed hydrocephaly and a high incidence of postnatal death on a C57BL/6 background. Cortical development proceeded normally, while hippocampus and hypothalamus relative brain volumes were altered. A hypoplastic anterior pituitary was also observed that likely contributes to the small size of the R342X mice. Behavior testing demonstrated deficits in associative learning, spatial memory and an anxiolytic phenotype. Taken together, the R342X mice represent a good preclinical model of BFLS that will allow further dissection of PHF6 function and disease pathogenesis.
PHF6 突变 c.1024C>T;p.R342X 是 Börjeson-Forssman-Lehmann 综合征(BFLS)的一个常见致病原因,BFLS 是一种神经发育障碍,其特征为中度至重度智力残疾、躯干肥胖、男性乳房发育、性腺功能减退、长而逐渐变细的手指和大耳朵(MIM#301900)。在这里,我们使用 CRISPR 技术生成了具有相同替换(R342X 小鼠)的转基因小鼠。我们表明,p.R342X 突变分别通过无意义介导的衰变和无意义相关的可变剪接导致 PHF6 蛋白水平在人和小鼠中降低。磁共振成像研究表明,R342X 小鼠在混合遗传背景下脑体积减小,但在 C57BL/6 遗传背景下发生脑积水和高比例的产后死亡。皮质发育正常进行,而海马体和下丘脑的相对脑体积发生改变。还观察到发育不全的垂体前叶,这可能导致 R342X 小鼠体积较小。行为测试表明,R342X 小鼠在联想学习、空间记忆和焦虑样表型方面存在缺陷。综上所述,R342X 小鼠代表了 BFLS 的一个很好的临床前模型,将允许进一步剖析 PHF6 的功能和疾病发病机制。