Amyloid deposition in a mouse model humanized at the transthyretin and retinol-binding protein 4 loci

Familial amyloidotic polyneuropathy is an autosomal dominant disorder caused by a point mutation in the transthyretin (TTR) gene. The process of TTR amyloidogenesis begins with rate-limiting dissociation of the TTR tetramer. Thus, the TTR stabilizers, such as Tafamidis and Diflunisal, are now in clinical trials. Mouse models will be useful to testing the efficacy of these drugs. Although several mouse models have been generated, they all express mouse Rbp4. Thus, human TTR associates with mouse RBP4, resulting in different kinetic and thermodynamic stability profiles of TTR tetramers. To overcome this problem, we previously produced humanized mouse strains at both the TTR and Rbp4 loci (Ttr ^hTTRVal30 , Ttr ^hTTRMet30 , and Rbp4 ^hRBP4 ). By mating these mice, we produced double-humanized mouse strains, Ttr ^{hTTRVal30/hTTRVal30} :Rbp4 ^hRBP4/hRBP4 and Ttr ^{hTTRVal30/Met30} :Rbp4 ^hRBP4/hRBP4 . We used conventional transgenic mouse strains on a wild-type (Ttr ^+/+ :Tg[6.0hTTRMet30]) or knockout Ttr background (Ttr^-/-:Tg[6.0hTTRMet30]) as reference strains. The double-humanized mouse showed 1/25 of serum hTTR and 1/40 of serum hRBP4 levels. However, amyloid deposition was more pronounced in Ttr ^{hTTRVal30/Met30} :Rbp4 ^hRBP4/hRBP4 than in conventional transgenic mouse strains. In addition, a similar amount of amyloid deposition was also observed in Ttr ^{hTTRVal30/ hTTRVal30} :Rbp4 ^{hRBP4/ hRBP4} mice that carried the wild-type human TTR gene. Furthermore, amyloid deposition was first observed in the sciatic nerve without any additional genetic change. In all strains, anti-TTR antibody-positive deposits were found in earlier age and at higher percentage than amyloid fibril deposition. In double-humanized mice, gel filtration analysis of serum revealed that most hTTR was free of hRBP4, suggesting importance of free TTR for amyloid deposition.