In utero therapy for hemophilia A using EGFP amniotic fluid stem cells

Abstract

Hemophilia A is an X chromosome recessive genetic disease, that causes clotting factor VIII (FVIII) deficiency leading to blood coagulation dysfunction. The activity of coagulation factor in serious hemophilia patients is less than one percent. The current therapeutic methods of hemophilia A are mainly for giving either the coagulation factor supplementary injection or gene therapy. However, both methods have some shortcomings. Therefore, an alternative potential treatment might be desired. The amniotic fluid stem cells (AFSCs) are derived from amniotic fluid. The amniotic fluid is the medical waste and it don't involve in ethical issues. Moreover, AFSCs can differentiate to diverse cells of ectoderm, mesoderm, and endoderm lineages. In utero transplantation (IUT) of AFSCs is an approach used to cure inherited genetic fetal defects during the gestation period of pregnant. In human, certain disease, such as osteogenesis imperfect, was successfully treated by using this method. By transplanting AFSCs into fetus, the AFSCs can directly involve in the development of the fetus and achieve therapeutic effect. The aim of this research is to assess therapeutic effect of hemophilia A after in utero transplantation by amniotic fluid‐derived EGFP‐stem cells (EGFP‐AFSCs) into the liver of FVIII knockout mice. In this study, the B6;129S‐F8tm1Kaz/J knockout mice was used as a hemophilia A animal model. The C57BL/6 mice was used as a normal control and untreated hemophilia A mice was used as a negative control. To establish IUT animal model, the pregnant CD‐1 mice (14.5 days) underwent abdominal surgery and transplanted EGFP‐AFSCs (5 × 104 cells per fetus) into the liver of fetus. Three weeks after birth, the mice were sacrificed and the EGFP signal in organs were analyzed by RT‐PCR, histological examination and flow cytometry. Results showed that EGFP signal can be detected in the liver and intestine. Furthermore, we used several methods including activated partial thromboplastin time, FVIII activity assay, tail clip survival rate, and hemoglobin concentration to analyze whether the FVIII activity in the hemophilia A mice can be restored or not after IUT operation by EGFP‐AFSCs. By conducting this research, we expect to solve the hemophilia patient's therapeutic problems and increase better quality of life.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.