Genetic modification of donor hepatocytes improves therapeutic efficacy for hemophilia B in mice.
Hepatocyte transplantation (Tx) holds promise for curing genetic liver diseases. However, a limited number of donor hepatocytes can be transplanted into the host liver. Recipient preconditioning and donor cell engineering are under investigation to improve cell engraftment. In theory, genetically engineered cells secreting therapeutic proteins with superior function could compensate for poor engraftment efficiency. We have generated a bioengineered human coagulation factor IX (FIX) with augmented specific activity (named FIX-Triple). The aim of this study was to evaluate therapeutic efficacy of cell therapy using hemophilia B (HB) as a disease model by transplanting FIX-Triple-secreting hepatocytes. The donor hepatocytes were isolated from FIX-Triple knock-in (KI) or FIX-WT (wild-type) KI mice and transplanted intrasplenically into FIX knock-out (KO) mice. FIX-Triple KI recipients exhibited fourfold higher plasma FIX clotting activity than FIX-WT KI recipients. By repeated Txs, the clotting activity of FIX-Triple KI recipients even increased to more than 10% of normal mouse plasma. The engraftment and FIX production efficiencies of transplanted cells were equivalent between the FIX-WT KI and FIX-Triple KI donors. A hemostatic function assay showed that FIX-Triple KI recipients with repeated Txs had more enhanced clot kinetics and a greater maximum rate of thrombus generation than those with a single Tx. Moreover, FIX inhibitors in these recipients rarely developed. In conclusion, hepatocyte Tx with genetically engineered donor cells is an effective therapeutic strategy for HB.
Wu
Yao-Ming
YM
Department of Surgery, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
Kao
Chung-Yang
CY
Huang
Yu-Jen
YJ
Yu
I-Shing
IS
Lee
Hsuan-Shu
HS
Lai
Hong-Shiee
HS
Lee
Po-Huang
PH
Lin
Chia-Ni
CN
Lin
Shu-Wha
SW
D000818:Animals; D017690:Cell Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D002836:Hemophilia B / Q000097:blood / Q000235:genetics / Q000601:surgery*; D022781:Hepatocytes / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation*; D006801:Humans; D008099:Liver / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D023822:Tissue Engineering / Q000379:methods
D005164:Factor IX