Heikkilä P, Tibell A, Morita T, Chen Y, Wu G, Sado Y, Ninomiya Y, Pettersson E, Tryggvason K
Division of Matrix Biology, Department of Medical Biochemistry and Biophysics.
Gene Ther. 2001 Jun;8(11):882-90. doi: 10.1038/sj.gt.3301342.
Gene therapy of Alport syndrome (hereditary nephritis) aims at the transfer of a corrected type IV collagen alpha chain gene into renal glomerular cells responsible for production of the glomerular basement membrane (GBM). A GBM network composed of type IV collagen molecules is abnormal in Alport syndrome which leads progressively to kidney failure. The most common X-linked form of the disease is caused by mutations in the gene for the alpha5(IV) chain, the alpha5 chain of type IV collagen. Full-length human alpha5(IV) cDNA was expressed in HT1080 cells with an adenovirus vector, and the recombinant alpha5(IV) chain was shown to assemble into heterotrimers consisting of alpha3(IV) and alpha4(IV) chains, utilizing a FLAG epitope in the recombinant alpha5(IV) chain. The results indicate that correction of the molecular defect in Alport syndrome is possible. Previously, we had developed an organ perfusion method for effective in vivo gene transfer into glomerular cells. In vivo perfusion of pig kidneys with the recombinant adenovirus resulted in expression of the alpha5(IV) chain in kidney glomeruli as shown by in situ hybridization and its deposition into the GBM was shown by immunohistochemistry. The results strongly suggest future possibilities for gene therapy of Alport syndrome.
奥尔波特综合征（遗传性肾炎）的基因治疗旨在将校正后的IV型胶原α链基因导入负责肾小球基底膜（GBM）产生的肾小球细胞。由IV型胶原分子组成的GBM网络在奥尔波特综合征中异常，这会逐渐导致肾衰竭。该疾病最常见的X连锁形式是由IV型胶原α5链（α5(IV)链）基因的突变引起的。全长人α5(IV) cDNA在HT1080细胞中用腺病毒载体表达，并且重组α5(IV)链被证明能与α3(IV)和α4(IV)链组装成异源三聚体，这利用了重组α5(IV)链中的一个FLAG表位。结果表明校正奥尔波特综合征中的分子缺陷是可能的。此前，我们开发了一种用于将基因有效体内导入肾小球细胞的器官灌注方法。用重组腺病毒对猪肾进行体内灌注，原位杂交显示α5(IV)链在肾小球中表达，免疫组织化学显示其沉积到GBM中。这些结果强烈提示了奥尔波特综合征基因治疗的未来可能性。
