1996 Fiscal Year Final Research Report Summary
Molecular Approach to an Amyloidosis by Using Gene Targeting
| Project/Area Number |
07044242
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Yamanashi Medical University |
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Principal Investigator |
MAEDA Shuichiro Yamanashi Medical University Professor, 医学部, 教授 (10117244)
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| Co-Investigator(Kenkyū-buntansha) |
EPISKOPOU Vasso Royal Postgraduate Medical School, Mouse Embryology, マウス発生学研究室, 室長
SARAIVA Maria.J.M Institute de Ciencias Biomedicas, Universidade do Porto, 医科学研究所, 教授
BUXBAUM J.N. ニューヨーク大学, 医学部, 教授
COSTANTINI F コロンビア大学, 医学部, 教授
GOTTESMAN M. コロンビア大学, 癌研究所, 所長
SHIMADA Kazunori Research Institute for Microbial Diseases, Osaka Universiy, 微生物病研究所, 教授 (40037354)
GOTTESMAN Max E Institute of Cancer Research, Columbia University
COSTANTINI Franklin D Department of Genetics and Development, Columbia University
BUXBAUM Joel N Laboratory of Molecular Pathogenesis, New York Department of Veterans Affairs Me
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| Project Period (FY) |
1995 – 1996
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| Keywords | amyloidosis / transthyretin / mouse model of human disease / familial amyloidotic polyneuropathy / gene targeting / embryonic Stem cells / serum amyloid P component / transgenic mouse |
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| Research Abstract |
1, The introduction of the human mutant transthyretin (ttr) gene (6.0-hMet 30) which is responsible for familial amyloidotic polyneuropathy (FAP) into the TTR-deficient (ttr^<-/->) mice significantly increased their depressed serum levels of thyroxine (T_4) and plasma retinol-binding protein (RBP), suggesting that human TTR Met 30 binds T_4 and RBP in vivo.The T_4-binding ability of human TTR Met 30 was confirmed by the analysis of T_4-binding proteins in the sera of ttr^<-/-> transgenic mice expressing 6.0-hMet 30. The T_4-binding studies also demonstrated the presence of hybrid tetramers between mouse and human TTR subunits in the ttr^<+/+> transgenic mice expressing 6.0-hMet 30.
2, The gene encoding serum amyloid P component (SAP) was disrupted in CCE embryonic stem (ES) cells by homologous recombination. After injection of the ES cell clones into C57BL/6 host blastocysts two germ-line chimeras were obtained. The chimeras were bred with C57BL/6 females and the mice heterozygous for t
… More
he disrupted sap allele were intercrossed. Genotyping of the resulting progeny of 30 different litters with an average litter size of 7 showed that live-born mice homozygous for the disrupted sap allele (sap^<-/->) were recovered at the predicted frequency. The absence of SAP in the serum of sap^<-/-> mice was confirmed by Western blot analysis. The SAP-deficient mice display no obvious phenotypic abnormalities and their fertility is normal. To investigate the relationship between SAP and amyloid deposition, we asked if experimental AA amyloidosis can be induced in the SAP-deficient mice. AA amyloid deposition occurred in the absence of SAP.We are now in the process of comparing the onset and progression of AA amyloid deposition in the sap^<-/-> and wild-type mice
3, Using a novel gene targeting procedure, we introduced an amyloidogenic point mutation (TTR Met 30) into the endogenous ttr gene in ES cells. After injection of the ES cell clones into C57BL/6 host blastocysts several chimeras were obtained. The chimeras are now being bred with C57BL/6 femals. Less
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[Publications] Nagata,Y.,Tashiro,F.,Yi,S.,Murakami,T.,Maeda,S.,Takahashi,K.,Shimada,K.,Okamura,H.and Yamamura,K.: "A 6-kb upstream region of the human transthyretin gene can direct developmental,tissue-specific,and quantitatively normal exprssion in transgenic mouse." J.Biochem.117. 169-175 (1995)
Description
「研究成果報告書概要(和文)」より
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[Publications] Wei,S.,Episkopou,V.,Piantedosi,R.,Maeda,S.,Shimada,K.,Gottesman,M.E.and Blaner,W.S.: "Studies on the metabolism of retinol and retinol-binding protein in transthyretin-deficient mice produced by homologous recombination." J.Biol.Chem.270. 866-870 (1995)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kohno K.,Palha J.A.,Miyakawa K.,Saraiva M.J.M.,Ito S.,Mabuchi T.,Blaner W.S.,Iijima H.,Tsukahara S.,Episkopou V.,Gottesman M.E.,Shimada K.,Takahashi K.,Yamamura K.,and Maeda S.: "Analysis of amyloid deposition in a transgenic mouse model of homozygous familial amyloidotic polyneuropathy." Am.J.Pathol. (印刷中). (1997)
Description
「研究成果報告書概要(和文)」より
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[Publications] Nagata, Y., Tashiro, F., Yi, S., Murakami, T., Maeda, S., Takahashi, K., Shimada, K., Okamura, H.and Yamamura, K.: "A 6-kb upstream region of the human transthyretin gene can direct developmental, tissue-specific, and quantitatively normal exprssion in transgenic mouse." J.Biochem.117. 169-175 (1995)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Wei, S., Episkopou, V., Piantedosi, R., Maeda, S., Shimada, K., Gottesman, M.E.and Blaner.W.S.: "Studies on the metabolism of retinol and retinol-binding protein in transthyretin-deficient mice produced by homologous recombination." J.Biol.Chem.270. 866-870 (1995)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kohno K., Palha J.A., Miyakawa K., Saraiva M.J.M., Ito S., Mabuchi T., Blaner W.S., Iijima H., Tsukahara S., Episkopou V., Gottesman M.E., Shimada K., Takahashi K., Yamamura K., and Maeda S.: "Analysis of amyloid deposition in a transgenic mouse model of homozygous familial amyloidotic polyneuropathy." Am.J.Pathol. (in press). (1997)
Description
「研究成果報告書概要(欧文)」より
