The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Department of Neurology, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Autophagy. 2022 Sep;18(9):2198-2215. doi: 10.1080/15548627.2022.2025570. Epub 2022 Mar 28.
FYCO1 (FYVE and coiled-coil domain containing 1) is an adaptor protein, expressed ubiquitously and required for microtubule-dependent, plus-end-directed transport of macroautophagic/autophagic vesicles. We have previously shown that loss-of-function mutations in cause cataracts with no other ocular and/or extra-ocular phenotype. Here, we show homozygous knockout () mice recapitulate the cataract phenotype consistent with a critical role of FYCO1 and autophagy in lens morphogenesis. Transcriptome coupled with proteome and metabolome profiling identified many autophagy-associated genes, proteins, and lipids respectively perturbed in mice lenses. Flow cytometry of (c.2206C>T) knock-in (KI) human lens epithelial cells revealed a decrease in autophagic flux and autophagic vesicles resulting from the loss of FYCO1. Transmission electron microscopy showed cellular organelles accumulated in (c.2206C>T) KI lens-like organoid structures and in mice lenses. In summary, our data confirm the loss of FYCO1 function results in a diminished autophagic flux, impaired organelle removal, and cataractogenesis. CC: congenital cataracts; DE: differentially expressed; ER: endoplasmic reticulum; FYCO1: FYVE and coiled-coil domain containing 1; hESC: human embryonic stem cell; KI: knock-in; OFZ: organelle-free zone; qRT-PCR: quantitative real-time PCR; PE: phosphatidylethanolamine; RNA-Seq: RNA sequencing; SD: standard deviation; sgRNA: single guide RNA; shRNA: shorthairpin RNA; TEM: transmission electron microscopy; WT: wild type.
FYCO1（FYVE 和卷曲螺旋结构域包含蛋白 1）是一种衔接蛋白，广泛表达，是微管依赖性、正极定向的大自噬/自噬囊泡运输所必需的。我们之前已经表明， 功能丧失突变导致白内障，没有其他眼部和/或眼外表型。在这里，我们展示 FYCO1 缺失纯合子敲除 () 小鼠重现白内障表型，这与 FYCO1 和自噬在晶状体形态发生中的关键作用一致。转录组与蛋白质组和代谢组谱分析分别鉴定了许多自噬相关基因、蛋白质和脂质在 小鼠晶状体中受到干扰。对 (c.2206C>T) 敲入 (KI) 人晶状体上皮细胞的流式细胞术显示，由于 FYCO1 的缺失，自噬通量和自噬囊泡减少。透射电子显微镜显示，细胞细胞器在 (c.2206C>T) KI 晶状体样类器官结构和 小鼠晶状体中积累。总之，我们的数据证实 FYCO1 功能的丧失导致自噬通量降低、细胞器清除受损和白内障形成。CC：先天性白内障；DE：差异表达；ER：内质网；FYCO1：FYVE 和卷曲螺旋结构域包含蛋白 1；hESC：人胚胎干细胞；KI：敲入；OFZ：细胞器自由区；qRT-PCR：实时定量 PCR；PE：磷脂酰乙醇胺；RNA-Seq：RNA 测序；SD：标准差；sgRNA：单引导 RNA；shRNA：短发夹 RNA；TEM：透射电子显微镜；WT：野生型。
