Pathogenomics of collagen disease using synthetic polymorphic proteins and BAG transgenic mice

使用合成多态蛋白和 BAG 转基因小鼠进行胶原病的病理基因组学

• 批准号: 13557018
• 负责人: NOSE Masato
• 金额: $ 8.58万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557018/
• 关键词: collagen discase; linkago analysis; collagen disease-susceptibility loci; protein synthesis in cell free system; synthetic polymorphic proteins; recombinant inbred strains of mice; BAC transgenic mice; pathogenomics; 自己抗体; ポリジーン; リンケージ解析; 位置的候補

Collagen disease involving SLE and RA shows a complex pathological manifestation such as glomerulonephritis, vasculitis, arthritis and/or sialoadenitis, which seems to result from the cumulative effect of multiple gene loci with an allelic combination. In this research, we focused on to identify these genes in MRL model mice.MRL/MpJ-lpr/lpr mice bearing a Fas deletion mutant gene lprr (MRL/lpr), spontaneously develop various forms of collagen disease in the same individuals, including glomerulonephritis, polyarteritis, arthritis and sialoadenitis, associated with an MRL background. Previously, we mapped susceptibility loci to each lesion with polymorphic microsatellite markers using N2 backcross and F2 intercross mice with C3H/lpr mice, and clarified that gene loci responsible for each lesion exist at different chromosomal positions and they have additive and hierarchical properties in a polygenic manner. To identify the candidate genes for each locus, we performed novel strategies. 1) functional analyses of synthetic polymorphic proteins corresponding to the candidate genes in vitro, 2) development of recombinant inbred strains of mice between MRL/lpr and C3H/lpr mice and the strain distribution pattern table, followed by the pathogenimics studies, 3) development of transgenic mice with BAG (bacterial artificial chromosomes) corresponding to the positional candidate loci, followed by the pathological analyses.We conclude that these studies were efficient to clarify the susceptibility genes to polygenic diseases involving collagen disease based on functional genomics.

涉及SLE和RA的胶原病表现为复杂的病理表现，如肾小球肾炎、血管炎、关节炎和/或涎腺炎，这似乎是多个基因座位和等位基因组合累积作用的结果。在本研究中，我们致力于在MRL模型小鼠中识别这些基因。携带Fas缺失突变基因LPR(MRL/LPR)的MRL/MPJ-LPR/LPR小鼠在同一个体中自发地发生各种形式的胶原病，包括肾小球肾炎、多动脉炎、关节炎和涎腺炎，与MRL背景相关。在此之前，我们利用N_2回交和F_2与C3H/LPR小鼠杂交，利用多态微卫星标记定位了每个病变的易感基因座，并阐明了每个病变的基因座存在于不同的染色体位置，它们以多基因的方式具有加性和等级性。为了确定每个基因座的候选基因，我们采取了新的策略。1)与候选基因相对应的人工合成多态蛋白的体外功能分析；2)在MRL/LPR和C3 H/LPR小鼠之间构建重组近交系小鼠及其菌株分布模式表，然后进行致病机理研究；3)建立与候选基因定位相对应的BAG(细菌人工染色体)转基因小鼠，并进行病理学分析。

项目成果

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Yuasa T、Ono M、Watanabe T、Takai F.：“Lyn 对于 Fcγ 受体 III 介导的全身性过敏反应至关重要，但对于 Arthus 反应则不然”J Exp Med 193. 563-571 (2001)

Takai T, Ono M.: "Activating and inhibitory nature of the murine paired immunoglobulin-like receptor family"Immunol Rev. 181. 215-222 (2001)

Takai T，Ono M.：“鼠配对免疫球蛋白样受体家族的激活和抑制性质”Immunol Rev. 181. 215-222 (2001)

Tsuneyama K, Nose M, Nisihara M, Katayanagi K, Harada K, Nakanuma Y: "Spontaneous occurrence of chronic non-suppurative destructive cholangitis and antimitochondrial autoantibodies in MRL/lpr mice : possible animal model for primary billary cirrhosis"Path

Tsuneyama K、Nose M、Nisihara M、Katayanagi K、Harada K、Nakanuma Y：“MRL/lpr 小鼠自发发生慢性非化脓性破坏性胆管炎和抗线粒体自身抗体：原发性胆汁性肝硬化的可能动物模型”路径

Sawasaki T, Ogasawara T, Morishita R, Endo Y.: "A cell-free protein synthesis system for high-throughput proteomics"Proc Natl Acad Sci U S A. 99. 4652-4657 (2002)

Sawasaki T、Ogasawara T、Morishita R、Endo Y.：“用于高通量蛋白质组学的无细胞蛋白质合成系统”Proc Natl Acad Sci U S A. 99. 4652-4657 (2002)

Xiu Y, Nakamura K, Abe M, Li N, Wen XS, Jiang Y, Zhang D, Tsurui H, Matsuoka S, Hamano Y, Fujii H, Ono M, Takai T, Shimokawa T, Ra C, Shirai T, Hirose S.: "Transcriptional regulation of Fcvgr2b gene by polymorphic promoter region and its contribution to h

Xiu Y, Nakamura K, Abe M, Li N, Wen XS, Jiang Y, 张 D, Tsurui H, Matsuoka S, Hamano Y, Fujii H, Ono M, Takai T, Shimokawa T, Ra C, Shirai T, Hirose S