背景:脂蛋白脂肪酶(LPL)在甘油三酯水解中起着至关重要的作用。导致功能完全或接近完全丧失的LPL基因中的罕见双等位基因变异导致常染色体隐性家族性乳糜微粒血症综合征。然而,罕见的双等位基因LPL变异导致显著但部分功能丧失的文献很少记载。这项研究报道了在怀孕期间患有高甘油三酯血症诱导的急性胰腺炎(HTG-AP)的中国患者中这种罕见的双等位基因LPL变体的新发生,并提供了深入的功能表征。
方法:LPL的完整编码序列和相邻内含子区域,通过Sanger测序分析APOC2、APOA5、LMF1和GPIHBP1基因。目的是识别罕见的变异,包括胡说八道,移码,错觉,小的框内缺失或插入,和典型的剪接位点突变。鉴定的LPL错义变体对蛋白质表达的功能影响,分泌,通过单次和共转染实验在HEK293T细胞中评估活性,有和没有肝素治疗。
结果:在患者中发现了两种罕见的LPL错义变异:先前报道的c.809G>A(p。Arg270His)和小说c.331G>C(p。Val111Leu)。遗传测试证实这些变体是双等位基因遗传的。功能分析显示,p.Arg270His变体由于对蛋白质合成/稳定性的影响而导致LPL功能几乎完全丧失,分泌,和酶活性。相比之下,p.Val111Leu变体保留了大约32.3%的野生型活性,在不影响蛋白质合成的情况下,稳定性,或分泌物。共转染实验表明,联合活性水平为20.7%,表明变体之间没有显性的负相互作用。患者的肝素后血浆LPL活性约为对照水平的35%。
结论:本研究提供了一个在妊娠期间患有HTG-AP的患者出现部分但显著的双等位基因LPL变异的新病例。我们的发现增强了对LPL基因型和临床表型之间细微差别关系的理解。强调残余LPL功能在疾病表现和严重程度中的重要性。此外,我们的研究强调了根据美国医学遗传学和基因组学学院(ACMG)的变异分类指南对经典孟德尔疾病基因中的部分功能缺失变异进行分类的挑战.
BACKGROUND: Lipoprotein lipase (LPL) plays a crucial role in triglyceride hydrolysis. Rare biallelic variants in the LPL gene leading to complete or near-complete loss of function cause autosomal recessive familial chylomicronemia syndrome. However, rare biallelic LPL variants resulting in significant but partial loss of function are rarely documented. This study reports a novel occurrence of such rare biallelic LPL variants in a Chinese patient with hypertriglyceridemia-induced acute pancreatitis (HTG-AP) during pregnancy and provides an in-depth functional characterization.
METHODS: The complete coding sequences and adjacent intronic regions of the LPL, APOC2, APOA5, LMF1, and GPIHBP1 genes were analyzed by Sanger sequencing. The aim was to identify rare variants, including nonsense, frameshift, missense, small in-frame deletions or insertions, and canonical splice site mutations. The functional impact of identified LPL missense variants on protein expression, secretion, and activity was assessed in HEK293T cells through single and co-transfection experiments, with and without heparin treatment.
RESULTS: Two rare LPL missense variants were identified in the patient: the previously reported c.809G > A (p.Arg270His) and a novel c.331G > C (p.Val111Leu). Genetic testing confirmed these variants were inherited biallelically. Functional analysis showed that the p.Arg270His variant resulted in a near-complete loss of LPL function due to effects on protein synthesis/stability, secretion, and enzymatic activity. In contrast, the p.Val111Leu variant retained approximately 32.3% of wild-type activity, without impacting protein synthesis, stability, or secretion. Co-transfection experiments indicated a combined activity level of 20.7%, suggesting no dominant negative interaction between the variants. The patient\'s post-heparin plasma LPL activity was about 35% of control levels.
CONCLUSIONS: This study presents a novel case of partial but significant loss-of-function biallelic LPL variants in a patient with HTG-AP during pregnancy. Our findings enhance the understanding of the nuanced relationship between LPL genotypes and clinical phenotypes, highlighting the importance of residual LPL function in disease manifestation and severity. Additionally, our study underscores the challenges in classifying partial loss-of-function variants in classical Mendelian disease genes according to the American College of Medical Genetics and Genomics (ACMG)\'s variant classification guidelines.