Abstract：
OBJECTIVE: Oocyte maturation defect (OOMD) is a rare cause of in vitro fertilization failure characterized by the production of immature oocytes. Compound heterozygous or homozygous PATL2 mutations have been associated with oocyte arrest at the germinal vesicle (GV), metaphase I (MI), and metaphase II (MII) stages, as well as morphological changes.
METHODS: In this study, we recruited three OOMD cases and conducted a comprehensive multiplatform laboratory investigation.
RESULTS: Whole exome sequence (WES) revealed four diagnostic variants in PATL2, nonsense mutation c.709C > T (p.R237*) and frameshift mutation c.1486_1487delinsT (p.A496Sfs*4) were novel mutations that have not been reported previously. Furthermore, the pathogenicity of these variants was predicted using in silico analysis, which indicated detrimental effects. Molecular dynamic analysis suggested that the A496S variant disrupted the hydrophobic segment, leading to structural changes that affected the overall protein folding and stability. Additionally, biochemical and molecular experiments were conducted on cells transfected with wild-type (WT) or mutant PATL2 (p.R237* and p.A496Sfs*4) plasmid vectors.
CONCLUSIONS: The results demonstrated that PATL2A496Sfs*4 and PATL2R237* had impacts on protein size and expression level. Interestingly, expression levels of specific genes involved in oocyte maturation and early embryonic development were found to be simultaneously deregulated. The findings in our study expand the variation spectrum of the PATL2 gene, provide solid evidence for counseling on future pregnancies in affected families, strongly support the application of in the diagnosis of OOMD, and contribute to the understanding of PATL2 function.
METHODS: In this study, we recruited three OOMD cases and conducted a comprehensive multiplatform laboratory investigation.
RESULTS: Whole exome sequence (WES) revealed four diagnostic variants in PATL2, nonsense mutation c.709C > T (p.R237*) and frameshift mutation c.1486_1487delinsT (p.A496Sfs*4) were novel mutations that have not been reported previously. Furthermore, the pathogenicity of these variants was predicted using in silico analysis, which indicated detrimental effects. Molecular dynamic analysis suggested that the A496S variant disrupted the hydrophobic segment, leading to structural changes that affected the overall protein folding and stability. Additionally, biochemical and molecular experiments were conducted on cells transfected with wild-type (WT) or mutant PATL2 (p.R237* and p.A496Sfs*4) plasmid vectors.
CONCLUSIONS: The results demonstrated that PATL2A496Sfs*4 and PATL2R237* had impacts on protein size and expression level. Interestingly, expression levels of specific genes involved in oocyte maturation and early embryonic development were found to be simultaneously deregulated. The findings in our study expand the variation spectrum of the PATL2 gene, provide solid evidence for counseling on future pregnancies in affected families, strongly support the application of in the diagnosis of OOMD, and contribute to the understanding of PATL2 function.
摘要：
目的：卵母细胞成熟缺陷（OOMD）是体外受精失败的罕见原因，其特征是未成熟卵母细胞的产生。复合杂合子或纯合子PATL2突变与卵母细胞在生发囊泡（GV）的停滞有关，中期I（MI），和中期II(MII)阶段，以及形态变化。
方法：在本研究中，我们招募了3例OOMD病例,并进行了全面的多平台实验室调查.
结果：整个外显子组序列（WES）揭示了PATL2中的四个诊断变体，无义突变c.709C>T（p。R237*）和移码突变c.1486_1487delinsT（p。A496Sfs*4）是以前没有报道的新突变。此外，使用计算机分析预测这些变异的致病性，这表明了有害的影响。分子动力学分析表明，A496S变体破坏了疏水片段，导致影响整体蛋白质折叠和稳定性的结构变化。此外,在用野生型（WT）或突变型PATL2转染的细胞上进行了生化和分子实验（p。R237*和p.A496Sfs*4)质粒载体。
结论：结果表明，PATL2A496Sfs*4和PATL2R237*对蛋白质大小和表达水平有影响。有趣的是,发现参与卵母细胞成熟和早期胚胎发育的特定基因的表达水平同时失调。我们的研究结果扩展了PATL2基因的变异谱，为受影响家庭的未来怀孕提供可靠的咨询证据，强烈支持在OOMD诊断中的应用，并有助于对PATL2函数的理解。
方法：在本研究中，我们招募了3例OOMD病例,并进行了全面的多平台实验室调查.
结果：整个外显子组序列（WES）揭示了PATL2中的四个诊断变体，无义突变c.709C>T（p。R237*）和移码突变c.1486_1487delinsT（p。A496Sfs*4）是以前没有报道的新突变。此外，使用计算机分析预测这些变异的致病性，这表明了有害的影响。分子动力学分析表明，A496S变体破坏了疏水片段，导致影响整体蛋白质折叠和稳定性的结构变化。此外,在用野生型（WT）或突变型PATL2转染的细胞上进行了生化和分子实验（p。R237*和p.A496Sfs*4)质粒载体。
结论：结果表明，PATL2A496Sfs*4和PATL2R237*对蛋白质大小和表达水平有影响。有趣的是,发现参与卵母细胞成熟和早期胚胎发育的特定基因的表达水平同时失调。我们的研究结果扩展了PATL2基因的变异谱，为受影响家庭的未来怀孕提供可靠的咨询证据，强烈支持在OOMD诊断中的应用，并有助于对PATL2函数的理解。
