Donnelly Centre, University of Toronto, Toronto, ON, Canada.
Department of Biochemistry, University of Toronto, Toronto, ON, Canada.
Mol Syst Biol. 2022 Feb;18(2):e10629. doi: 10.15252/msb.202110629.
Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride and bicarbonate channel in secretory epithelia with a critical role in maintaining fluid homeostasis. Mutations in CFTR are associated with Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder in Caucasians. While remarkable treatment advances have been made recently in the form of modulator drugs directly rescuing CFTR dysfunction, there is still considerable scope for improvement of therapeutic effectiveness. Here, we report the application of a high-throughput screening variant of the Mammalian Membrane Two-Hybrid (MaMTH-HTS) to map the protein-protein interactions of wild-type (wt) and mutant CFTR (F508del), in an effort to better understand CF cellular effects and identify new drug targets for patient-specific treatments. Combined with functional validation in multiple disease models, we have uncovered candidate proteins with potential roles in CFTR function/CF pathophysiology, including Fibrinogen Like 2 (FGL2), which we demonstrate in patient-derived intestinal organoids has a significant effect on CFTR functional expression.
囊性纤维化跨膜电导调节因子（CFTR）是分泌上皮细胞中的氯离子和碳酸氢根离子通道，在维持液体动态平衡中起着关键作用。CFTR 的突变与囊性纤维化（CF）有关，CF 是白种人中最常见的致命常染色体隐性遗传病。虽然最近以直接修复 CFTR 功能障碍的调节剂药物的形式取得了显著的治疗进展，但仍有很大的空间可以提高治疗效果。在这里，我们报告了高通量筛选变体哺乳动物膜双杂交（MaMTH-HTS）的应用，以绘制野生型（wt）和突变 CFTR（F508del）的蛋白质-蛋白质相互作用图谱，努力更好地了解 CF 细胞效应并为患者特异性治疗确定新的药物靶点。结合在多种疾病模型中的功能验证，我们已经发现了一些候选蛋白，它们可能在 CFTR 功能/ CF 病理生理学中发挥作用，包括纤维蛋白原样 2（FGL2），我们在患者来源的肠类器官中证明，它对 CFTR 功能表达有显著影响。
