MAKING OF MOUSE MODEL OF ADPKD AND GENE THERAPY

Research Project

Project/Area Number	10671462
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Urology
Research Institution	The University of Tokyo, Branch Hospital
Principal Investigator	HORIE Shigeo UNIVERSITY OF TOKYO BRANCH HOSPITAL, ASSISTANT PROFES., 医学部・附属病院分院, 講師 (40190243)
Co-Investigator(Kenkyū-buntansha)	AIBA Atsu MEDICAL SCIENCE CENTER, IMSUT, UNIVERSITY OF TOKYO, ASSOCIATE PROFES., 医科学研究所・ヒト疾患モデル研究センター, 助教授 (20271116)
Project Period (FY)	1998 – 1999
Project Status	Completed (Fiscal Year 1999)
Budget Amount *help	¥1,600,000 (Direct Cost: ¥1,600,000) Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000) Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywords	ADPKD / Polycystin 2 / ES cell / 多発性嚢胞腎 / Polycystin1 / ターゲッティング / ノックアウトマウス / PKD2 / ジーンターゲッティング
Research Abstract	The majority of autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations to either the PKD1 or PKD2 gene. Recent studies suggest that signaling would occur by the interaction between polycystin-1 and polycystin-2. Previously reported targeted mutation of Pkd1, the mouse homologue of PKD1 (Nat Genet. 17 : 179, Proc Natl Acad Sci USA.97 : 1731), disrupted the middle of 3'end of the gene. They revealed that truncated polycystin-1 is required in maintaining the structural integrity of the epithelium and vasculature and that and that truncated product may have a detrimental effect. We have generated mice carrying a mutation at the 5'end of the Pkd1 by gene targeting. Exons 2-6 of Pkd1 Were replaced by the neo cassette in the E14 line of ES cells. Two independent ES clones heterozygous for the mutant allele were identified, and injected into blastocysts from C57B6 mice. The male chimeras were mated with normal females to generate heterozgous mice. Embryos homozygous for ti … More me mutant allele (Pkd1^<-1->) showed cardiac defects as double-outletc right ventricle and ventricle septum defects at embryonic day 12.5 (E12.5). Abnormal faces and focal hemorrhage were observed in some embryos. Profound subculaneous edema and the increase in the amount of amniotic fluid were exhibited at E13.5 or later. Renal cysts appeared at E14.5. Pkd1-/- mice die in utero between E13.5 and 16.5. Adult Pkd1- heterozygous (+/-) mice developed renal cysts by 8 months, which was earlier than reported in the del34 mice. These phenotypes were similar to those of Pkd2^<-1-> mice (Nat Genet. 24 : 75), which would provide evidence that polycystin-1 and -2 share the common signaling and that polycystin-1 is expressed in neural-crest derived tissue. The cardiac defects and abnormal faces observed in these mice may have a common etiology that is based on the effect of polycystin-1 on neural crest cells. Conclusions : Our results indicate that signaling mediated by the interaction of polycystin-1 and -2 plays a critical role for normal cardiac development. Less