Hitomi Masahiro, Venegas Juan, Kang Shin Chung, Eng Charis
Genomic Medicine Institute, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, 44195, USA.
Department of Molecular Medicine, The Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, OH, 44195, USA.
Oncogene. 2023 Dec;42(50):3698-3707. doi: 10.1038/s41388-023-02867-4. Epub 2023 Nov 1.
Individuals with a PTEN germline mutation receive the molecular diagnosis of PTEN hamartoma tumor syndrome (PHTS). PHTS displays a complex spectrum of clinical phenotypes including harmartomas, predisposition to cancers, and autism spectrum disorder (ASD). Clear-cut genotype-phenotype correlations are yet to be established due to insufficient information on the PTEN function being impacted by mutations. To fill this knowledge gap, we compared functional impacts of two selected missense PTEN mutant alleles, G132D and M134R, each respectively being associated with distinct clinical phenotype, ASD or thyroid cancer without ASD using gene-edited human induced pluripotent stem cells (hiPSCs). In homozygous hiPSCs, PTEN expression was severely reduced by M134R mutation due to shortened protein half-life. G132D suppressed PTEN expression to a lesser extent than Μ134R mutation without altering protein half-life. When challenged with γ-irradiation, G132D heterozygous cells exited radiation-induced G2 arrest earlier than wildtype and M134R heterozygous hiPSCs despite the similar DNA damage levels as the latter two. Immunoblotting analyses suggested that γ-irradiation induced apoptosis in G132D heterozygous cells to lesser degrees than in the hiPSCs of other genotypes. These data suggest that ASD-associated G132D allele promotes genome instability by premature cell cycle reentry with incomplete DNA repair.
携带PTEN种系突变的个体被诊断为PTEN错构瘤肿瘤综合征(PHTS)。PHTS表现出一系列复杂的临床表型,包括错构瘤、癌症易感性和自闭症谱系障碍(ASD)。由于关于PTEN功能受突变影响的信息不足,尚未建立明确的基因型-表型相关性。为了填补这一知识空白,我们使用基因编辑的人类诱导多能干细胞(hiPSC)比较了两个选定的错义PTEN突变等位基因G132D和M134R的功能影响,每个等位基因分别与不同的临床表型、ASD或无ASD的甲状腺癌相关。在纯合hiPSC中,由于蛋白质半衰期缩短,M134R突变导致PTEN表达严重降低。G132D对PTEN表达的抑制程度低于M134R突变,且不改变蛋白质半衰期。当受到γ射线照射时,尽管G132D杂合细胞的DNA损伤水平与野生型和M134R杂合hiPSC相似,但它们比后两者更早地退出辐射诱导的G2期阻滞。免疫印迹分析表明,γ射线照射诱导G132D杂合细胞凋亡的程度低于其他基因型的hiPSC。这些数据表明,与ASD相关的G132D等位基因通过不完全DNA修复导致的过早细胞周期重新进入促进基因组不稳定。