Conventional therapies for hemophilia A(HA)are prophylactic or on-demand intravenous FⅧinfusions.However,they are expensive and inconvenient to perform.Thus,better strategies for HA treatment must be developed.In this study,a recombinant FⅧ cDNA encoding a human/rat hybrid FⅧ with an enhanced procoagulant potential for adeno-associated virus(AAV)-delivered gene therapy was developed.Plasmids containing human FⅧ heavy chain(hHC),human light chain(hLC),and rat light chain(rLC)were transfected into cells and hydrodynamically injected into HA mice.Purified AAV viruses were intravenously injected into HA mice at two doses.Results showed that the hHC+rLC protein had a higher activity than the hHC+hLC protein at comparable expression levels.The specific activity of hHC+rLC was about 4-to 8-fold higher than that of their counterparts.Hydrodynamic injection experiments obtained consistent results.Notably,the HA mice undergoing the AAV-delivered hHC+rLC treatment exhibited a visibly higher activity than those treated with hHC+hLC,and the therapeutic effects lasted for up to 40 weeks.In conclusion,the application of the hybrid FⅧ(hHC+rLC)via an AAV-delivered gene therapy substantially improved the hemorrhagic diathesis of the HA mice.These data might be of help to the development of optimized FⅧ expression cassette for HA gene therapy.

Jianhua Mao;Yun Wang;Wei Zhang;Yan Shen;Guowei Zhang;Wenda Xi;Qiang Wang;Zheng Ruan;Jin Wang;Xiaodong Xi

Shanghai Institute of Hematology,State Key Laboratory of Medical Genomics,Collaborative Innovation Center of Hematology,Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;Shanghai Institute of Hematology,State Key Laboratory for Medical Genomics and Department of Hematology,Collaborative Innovation Center of Systems Biomedicine,P?le Sino-Fran?ais des Sciences du Vivant et Genomique,Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;Research Center for Experimental Medicine,Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;The School of Medicine,Hangzhou Normal University,Hangzhou 310036,China;Shanghai Institute of Hypertension,Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China

医学前沿

[1]Jianhua Mao;Yun Wang;Wei Zhang;Yan Shen;Guowei Zhang;Wenda Xi;Qiang Wang;Zheng Ruan;Jin Wang;Xiaodong Xi-.Long-term correction of hemorrhagic diathesis in hemophilia A mice by an AAV-delivered hybrid FⅧ composed of the human heavy chain and the rat light chain)[J].医学前沿,2022(04):584-595

diathesis,procoagulant,hHC,hLC,rLC,hydrodynamically,Purified,hHC+rLC,hHC+hLC

Long,term,correction,hemorrhagic,hemophilia,mice,by,AAV,delivered,hybrid,composed,human,heavy,chain,light,Conventional,therapies,HA,are,prophylactic,demand,infusions,However,they,expensive,inconvenient,perform,Thus,better,strategies,treatment,must,developed,In,this,study,recombinant,cDNA,encoding,enhanced,potential,adeno,associated,gene,therapy,was,Plasmids,containing,were,transfected,into,cells,injected,viruses,intravenously,two,doses,Results,showed,that,protein,had,higher,activity,than,comparable,expression,levels,specific,about,fold,their,counterparts,Hydrodynamic,injection,experiments,obtained,consistent,results,Notably,undergoing,exhibited,visibly,those,treated,therapeutic,effects,lasted,up,weeks,conclusion,application,via,substantially,improved,These,data,might,help,development,optimized,cassette

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