Molecular and clinical characterization of two independent Chinese families with protein C deficiency

doi:10.1007/s00277-024-06156-2

科研成果详情

题名	Molecular and clinical characterization of two independent Chinese families with protein C deficiency
作者	Wen, Mengzhen; Lu, Yifan; Xie, Haixiao; Qin, Langyi; Ye, Longying; Zhang, Ke; Wang, Mingshan; Yang, Lihong
发表日期	2024-12-26
发表期刊	ANNALS OF HEMATOLOGY 影响因子和分区
语种	英语
原始文献类型	Article ; Early Access
关键词	Protein C deficiency Novel mutation Bioinformatics Thrombin generation assay In vitro expression
其他关键词	PLASMA
摘要	This study aims to investigate the clinical characterization and molecular pathogenic basis of hereditary protein C (PC) deficiency in two independent Chinese families, and conduct in vitro expression studies on the newly discovered p.Trp444Arg mutation. The PC activity (PC: A) was tested using the chromogenic substrate, and PC antigen (PC: Ag) was detected via enzyme-linked immunosorbent assay (ELISA). To identify the mutation sites, nine exons of the PROC gene were amplified by PCR, and the products were directly sequenced. The conservation and pathogenicity of the mutations, as well as changes in the spatial structure of PC proteins before and after mutations, were analyzed using ClustalX-2.1-win, online bioinformatics software, and PyMOL. The function of the mutant proteins was detected using the thrombin generation assay (TGA). Recombinant PC was ectopically expressed in HEK293T cells, with mRNA levels quantified by RT-qPCR. The recombinant protein was further characterized using Western blotting, ELISA, and immunofluorescence microscopy. Proband A and B, aged 39 and 63 respectively, are both diagnosed with deep vein thrombosis (DVT) in both lower limbs and pulmonary embolism (PE). Two missense mutations, p.Arg440Cys and p.Trp444Arg, were identified in the probands. Bioinformatics and protein modeling analyses revealed that the two mutations probably affected the normal function of PC. The thrombin generation assay revealed impaired thrombin generation capacity in both probands, with proband B showing more severe impairment. In vitro expression experiments demonstrated that p.Trp444Arg do not significantly affect mRNA expression levels of PC protein compared to wild-type, but result in lower PC: Ag content and protein expression in the supernatant and higher levels in the lysate. These two mutations may be the causes of reduced PC in two independent Chinese families. Notably, this is the first reported instance of the p.Trp444Arg mutation.
资助项目	Wenzhou Basic Medical and Health Science and Technology Project
出版者	SPRINGER
ISSN	0939-5555
EISSN	1432-0584
DOI	10.1007/s00277-024-06156-2
页数	10
WOS类目	Hematology
WOS研究方向	Hematology
WOS记录号	WOS:001383469200001
收录类别	SCIE ; PUBMED
URL	查看原文
PubMed ID	39724247
通讯作者地址	[Yang, Lihong]Wenzhou Med Univ, Dept Clin Lab, Key Lab Clin Lab Diag & Translat Res Zhejiang Prov, Affiliated Hosp 1, Wenzhou 325000, Peoples R China.
引用统计
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/225001
专题	附属第一医院
通讯作者	Yang, Lihong
作者单位	Wenzhou Med Univ, Dept Clin Lab, Key Lab Clin Lab Diag & Translat Res Zhejiang Prov, Affiliated Hosp 1, Wenzhou 325000, Peoples R China
第一作者单位	附属第一医院
通讯作者单位	附属第一医院
第一作者的第一单位	附属第一医院
推荐引用方式 GB/T 7714	Wen, Mengzhen,Lu, Yifan,Xie, Haixiao,et al. Molecular and clinical characterization of two independent Chinese families with protein C deficiency[J]. ANNALS OF HEMATOLOGY,2024.
APA	Wen, Mengzhen., Lu, Yifan., Xie, Haixiao., Qin, Langyi., Ye, Longying., ... & Yang, Lihong. (2024). Molecular and clinical characterization of two independent Chinese families with protein C deficiency. ANNALS OF HEMATOLOGY.
MLA	Wen, Mengzhen,et al."Molecular and clinical characterization of two independent Chinese families with protein C deficiency".ANNALS OF HEMATOLOGY (2024).