Examination of Gene Therapy by Bone Marrow Derived Stem Cells to Alport Syndrome

骨髓干细胞对阿尔波特综合征的基因治疗研究

• 批准号: 16390296
• 负责人: UCHIYAMA Makoto
• 金额: $ 4.86万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390296/
• 关键词: Alport syndrome; Alport syndrome model mice; bone marrow transplantation; gene therapy; Alport症候群; Col4a4ノックアウトマウス; トランスジェニックマウス

1.Examination of gene therapy by transplantating bone marrow derived stem cells of GFP mouse to Col4a4 knockout mice(1)We demonstrated that allogeic bone marrow transplatation into irradiated COL4a4 knockout mice prepared as autosomal chromosome type Alport syndrome model mice by COL4A4 mutation, leaded, to recruitment of resident glomerular cells. In the present study, transplantation of green fluorescent protein (GFP) mouse's BM into 4-week-old and 8-week-old Col4a4 knockout mice leaded to recruitment of resident glomerular cells at 12 weeks of age.(2)Type IV collagen a3 (COL4A3), a4 (COL4A4) and a5 (COLA5) chains were not detected in the glomerular basement membrane (GBM) after BMT by the immunohistochemistry.2.Generating the transgenic mice overexpressing type IV collagen a3,4, and a5 chains in the bone marrow stem cells and transplantion of the bone marrow derived stem cell of the transgenic mice to Col4a4 knockout mice(1)We generated the transgenic mice overexpressing type IV collagen a3,4, and a5 chains in the bone marrow cells. The survival and growth of the transgenic mice are not different from those of wild-type mice at present.(2)Type IV collagen a3,a4 and a5 chains were not only detected in the GBM of the Col4a4 knockout mouse by the bone marrow transplantation of the GFP mouse but also the transgenic mouse's BMT.3.ConclusionsType IV collagen a3,a4 and a5 chains were not detected in the GBM after BMT. The bone marrow derived stem cells could recruit as endothelial cells and mesangial cells, but couldn't recruit as podocytes. Therefore it is thought that only podcytes may form the a3- a4- a5(IV) network in the GBM, or BMT leads to recruitment of very few glomerular cells.

1.将GFP小鼠的骨髓源性干细胞移植到Col 4a 4基因敲除小鼠的基因治疗的检验（1）我们证明，将同种异体骨髓移植到通过COL 4A 4突变制备的常染色体型Alport综合征模型小鼠的经辐射的COL 4a 4基因敲除小鼠中，导致了固有肾小球细胞的募集。在本研究中，将绿色荧光蛋白（GFP）小鼠的BM移植到4周龄和8周龄的Col 4a 4基因敲除小鼠中，导致12周龄的常驻肾小球细胞的募集。（2）通过免疫组化检测，在骨髓移植后的肾小球基底膜（GBM）中未检测到IV型胶原α 3（COL 4A 3）、α 4（COL 4A 4）和α 5（COLA 5）链。骨髓干细胞中的α 4和α 5链以及将转基因小鼠的骨髓来源的干细胞移植到Col 4a 4敲除小鼠（1）我们产生了在骨髓细胞中过表达IV型胶原α 3、4和α 5链的转基因小鼠。目前，转基因小鼠的存活和生长与野生型小鼠没有区别。（2）Col 4a 4基因敲除小鼠骨髓移植后的GBM中可检测到Ⅳ型胶原α 3、α 4和α 5链，转基因小鼠骨髓移植后的GBM中也可检测到Ⅳ型胶原α 3、α 4和α 5链。骨髓源性干细胞可募集为内皮细胞和系膜细胞，但不能募集为足细胞。因此，认为只有足细胞可以在GBM中形成α 3-α 4-α 5（IV）网络，或者BMT导致非常少的肾小球细胞的募集。