Characterization of a genetically engineered mouse model of hemophilia A with complete deletion of the F8 gene.
Characterization of a genetically engineered mouse model of hemophilia A with complete deletion of the F8 gene.
复制标题
DOI：
10.1111/jth.13202
复制
发表时间：
2016-02
期刊：
影响因子：
--
通讯作者：
Lenardo MJ
中科院分区：
文献类型：
--
作者：
Chao BN;Baldwin WH;Healey JF;Parker ET;Shafer-Weaver K;Cox C;Jiang P;Kanellopoulou C;Lollar P;Meeks SL;Lenardo MJ
A型血友病治疗中最重要的并发症是患者在FVIII治疗后出现抑制性抗因子VIII （FVIII）抗体。不表达内源性FVIII即交叉反应物质（CRM）的严重血友病患者抑制剂形成的发生率最高。然而，目前的严重血友病A小鼠模型产生低水平的截断FVIII。缺乏相应的小鼠模型阻碍了在完全缺乏FVIII蛋白时抑制剂形成的研究。我们旨在建立一种新的严重血友病a小鼠模型（命名为F8TKO菌株）并对其进行表征，该模型缺乏F8和任何FVIII CRM的完整编码序列。以C57BL/6为背景，通过Cre-Lox重组构建小鼠，并通过体内出血试验、凝血和显色试验测量FVIII活性和ELISA检测抗FVIII抗体产生来对小鼠进行鉴定。F8TKO小鼠基因组中所有F8外显子编码区均被删除，未检测到F8 mRNA。F8TKO小鼠的出血表型与E16小鼠相似。F8TKO和E16小鼠注射重组FVIII后抗FVIII抗体滴度相近。我们描述了一种新的C57BL/6小鼠模型，用于缺乏CRM的严重血友病a患者。这些小鼠可以直接繁殖到许多C57BL/6基因工程小鼠，这对于在确定的遗传背景下研究多种基因对FVIII抑制剂形成的影响是有价值的。
The most important complication in hemophilia A treatment is the development of inhibitory anti-Factor VIII (FVIII) antibodies in patients after FVIII therapy. Patients with severe hemophilia who express no endogenous FVIII, i.e. cross-reacting material (CRM), have the greatest incidence of inhibitor formation. However, current mouse models of severe hemophilia A produce low levels of truncated FVIII. The lack of a corresponding mouse model hampers the study of inhibitor formation in the complete absence of FVIII protein. We aimed to generate and characterize a novel mouse model of severe hemophilia A (designated the F8TKO strain) lacking the complete coding sequence of F8 and any FVIII CRM. Mice were created on a C57BL/6 background using Cre-Lox recombination and characterized using in vivo bleeding assays, measurement of FVIII activity by coagulation and chromogenic assays, and anti-FVIII antibody production using ELISA. All F8 exonic coding regions were deleted from the genome and no F8 mRNA was detected in F8TKO mice. The bleeding phenotype of F8TKO mice was comparable to E16 mice by measurements of factor activity and tail snip assay. Similar levels of anti-FVIII antibody titers after recombinant FVIII injections were observed between F8TKO and E16 mice. We describe a new C57BL/6 mouse model for severe hemophilia A patients lacking CRM. These mice can be directly bred to the many C57BL/6 strains of genetically engineered mice making it valuable for studying the impact of a wide variety of genes on FVIII inhibitor formation on a defined genetic background.