未经证实:Bruck综合征(BS)是一种罕见的常染色体隐性遗传性成骨不全症,其特征是骨脆性和关节挛缩增加。I型BS的致病基因是FKBPl0,而II型BS的致病基因是PLOD2。两种类型的BS之间的临床表型没有发现显着差异。在这项研究中,我们对一个由PLOD2变异体引起的BS谱系进行了遗传分析,并研究了相应的细胞功能.
未经批准:血清生物化学,甲状旁腺激素(PTH),25-羟基维生素D[25-(OH)D],骨钙蛋白,并评估了患有BS的家庭成员的24小时尿钙水平。通过第二代测序和外显子捕获技术分析了先证者的基因。还对家族成员的可疑负责变体进行Sanger测序。通过基因克隆构建野生型和变异型慢病毒质粒并转染到HEK293T细胞中。通过实时定量聚合酶链反应验证细胞功能,西方印迹,和免疫荧光检测。
未经证实:在这个谱系中,发现先证者具有纯合变体c.1856G>A(p。Arg619His)在PLOD2(NM_182943.3)的外显子17中。他的近亲父母和姐妹是p.Arg619他的杂合携带者。构建的p.Arg619His变异细胞中PLOD2的mRNA表达显著上调,而细胞裂解物中PLOD2和I型胶原蛋白的表达显著下调。免疫荧光显示野生型PLOD2主要位于细胞质中,c.1856G>A变体后PLOD2蛋白的表达显著下调,几乎没有表情,与蛋白质印迹结果对齐。血清钠,钾,钙,磷,镁,碱性磷酸酶,PTH,25-(OH)D,骨钙蛋白,和24小时先证者的尿钙水平,他的父母,姐妹们都很正常.
未经授权:通过基因和细胞功能分析,初步证实PLOD2Arg619His错义变体通过减少蛋白质表达而引起BS。
UNASSIGNED: Bruck syndrome (BS) is a rare autosomal recessive inherited osteogenesis imperfecta disease characterized by increased bone fragility and joint contracture. The pathogenic gene of type I BS is FKBPl0, whereas that of type II BS is PLOD2. No significant difference has been found in the clinical phenotype between the two types of BS. In this
study, we performed genetic analysis of a BS pedigree caused by PLOD2 variant and studied the corresponding cellular function.
UNASSIGNED: Serum biochemistry, parathyroid hormone (PTH), 25-hydroxyvitamin D [25-(OH) D], osteocalcin, and 24-h urinary calcium levels of a family member with BS was assessed. The genes of the proband were analyzed by second-generation sequencing and exon capture techniques. Sanger sequencing was also performed for the suspected responsible variant of the family member. Wild- and variant-type lentivirus plasmids were constructed by gene cloning and transfected into HEK293T cells. Cell function was verified by real-time quantitative polymerase chain reaction, western blotting, and immunofluorescence detection.
UNASSIGNED: In this pedigree, the proband was found to have a homozygous variant c.1856G > A (p.Arg619His) in exon 17 of PLOD2 (NM_182943.3). His consanguineous parents and sisters were p.Arg619His heterozygous carriers. The mRNA expression of PLOD2 in the constructed p.Arg619His variant cells was significantly upregulated, while the expression of PLOD2 and collagen I protein in the cell lysate was significantly downregulated. Immunofluorescence revealed that the wild-type PLOD2 was mainly located in the cytoplasm, and the expression of the PLOD2 protein after c.1856G > A variant was significantly downregulated, with almost no expression, aligning with the western blot results. The serum sodium, potassium, calcium, phosphorus, magnesium, alkaline phosphatase, PTH, 25-(OH) D, osteocalcin, and 24 h urinary calcium levels of the proband, his parents, and sisters were normal.
UNASSIGNED: Through gene and cell function analyses, PLOD2 Arg619His missense variant was preliminarily confirmed to cause BS by reducing protein expression.