Factor XIII A subunit-deficient mice developed severe uterine bleeding events and subsequent spontaneous miscarriages

Abstract

IntroductionCoagulation factor XIII (FXIII) is a heterotetramer consisting of A subunits (FXIIIA) and B subunits (FXIIIB). FXIII catalyzes intermolecular cross-linking reactions between fibrin monomers, ␣ 2 -plasmin inhibitor, and fibronectin. These reactions increase the mechanical strength of the fibrin clot and its resistance to proteolytic degradation and enhance the assembly of the extracellular matrix. Accordingly, a deficiency of FXIII results in defective cross-linking reactions of these substrates and is thus associated with a number of diseases. 1 Congenital FXIII deficiency is caused by defects in the F13A or F13B genes, 2 leading to a bleeding tendency and abnormal wound healing in affected patients and spontaneous miscarriage in female patients. [3][4][5] To understand the molecular pathology of these deficiencies, we have identified a number of mutations in the F13A and F13B genes in patients' DNA and have analyzed the molecular mechanisms of FXIII deficiency using in vitro procedures. 6-15 Because it is not possible to understand completely the clinical pathologic mechanisms of this disease in vivo, we performed functional analyses on FXIIIA knockout (KO) mice. Study designFXIIIA KO mice were created by replacing exon VII with a neomycinresistance cassette as described previously. 16 All KO and wild-type CBA mice were housed in a specific pathogen-free facility. Experimental procedures were approved by the Animal Care and Use Committee of Yamagata University and were carried out in accordance with the guidelines of this committee and that of Japanese governmental law.Using tail biopsy DNA, genotyping of the mice was performed by polymerase chain reaction (PCR) with 2 forward primers designed from the neomycin-resistant gene (5Ј-CAC TGC ATT CTA GTT GTG GTT TGT CC-3Ј) and exon VII of the mouse F13A gene (5Ј-GCC AAG GAT GAT GAA GGT GTT CTT-3Ј), respectively, and a common reverse primer from intron G (5Ј-CCC TGA GAC TTA CGG ATG AAG AAG-3Ј).Tissues collected for histologic analyses were placed in 10% neutralbuffered formalin, processed into paraffin, and 3-m sections were prepared and stained with hematoxylin and eosin by standard techniques.The distribution of events between the study groups was analyzed using 4 ϫ 2 contingency tables for individual study groups as well as 2 ϫ 2 tables for a combined group of female wild-type mice and female homozygous FXIIIA KO mice. The 2 test was also carried out by using StatView software (Abacus Concepts, Berkeley, CA). Differences were considered statistically significant at P values less than .05. Results and discussionWestern blot analysis and amine-incorporation enzymatic assay confirmed the complete absence of FXIIIA in the uterus and plasma (where FXIIIA is normally expressed/present) of the homozygous FXIIIA KO mice identified by genotyping (data not shown). Despite the complete loss of FXIIIA, all homozygous FXIIIA KO mice had appeared normal at birth as previously reported. 16 Whereas all 20 of the female homozygous KO mice survived, 3 of 20 male KO mice...

Factor XIII Deficiency Causes Cardiac Rupture, Impairs Wound Healing, and Aggravates Cardiac Remodeling in Mice With Myocardial Infarction

“…Breeding of these mice is difficult because of intrauterine bleeding, with a resulting maternal mortality rate of 50% 9 ; however, the feasibility of obtaining up to 2.2 pups per homozygous breeding pair has been reported. 11 Mice underwent left coronary artery ligation or sham surgery as described previously. 12 Sham surgery comprised thoracotomy and passing of a suture under the coronary artery without actual ligation of the artery.…”

Roles of transglutaminases in cardiac and vascular diseases

“…TG2 not only enhances signaling of subtypes of alpha 1 -AR, but also thromboxane A2 receptor α subtype 135 and the oxytocin receptor (136,137). TG2 also associates with the cytoplasmic tails of several integrins including alpha 5 , alpha v , and alpha IIb (138).…”

“…Human Factor XIII deficiency is characterized by a life-long severe bleeding disorder, with intracranial hemorrhage (2) and recurrent abortions (3) being a significant risk. Factor XIII subunit A−/− mice have a similar phenotype with impaired clot formation and reduced clot stability (4), and also suffer from spontaneous miscarriages due to uterine bleeding (5).…”

Transglutaminases and Disease: Lessons From Genetically Engineered Mouse Models and Inherited Disorders

“…Further studies with TG2 In women, habitual abortion, as in afibrinogenemia and in some cases of hypodysfibrinogenemia, is a frequent manifestation of FXIII deficiency (170). Similarly, mixed strain (SVJ129-C57BL/6-Swiss albino-129O1a-CBACa) FXIII-A Ϫ/Ϫ mice are fertile, but reproduction is greatly impaired (148), with female knockout animals frequently developing overt genital bleeding around the 10th day of gestation (regardless of the genotype of the male mating partners), resulting in fetal abortion. About 50% of pregnancies result in maternal death.…”