Discovery of novel compounds regulating traffic to lysosomes and application for development of therapeutics

我们对溶酶体β-己糖胺酶(Hex)缺陷包括Tay-Sachs病(TSD)和Sandhoff病(SD)的细胞内交通异常进行了研究，以开发新的酶替代疗法，该疗法基于在细胞外给药时促进重组Hex向溶酶体输送的技术。我们开发了高含量甘露糖6-磷酸(M6P)残基的修饰Hex，通过细胞表面M6P受体有效地掺入了疾病模型培养细胞。此外，我们建立了一株来源于TSD患者的iPS细胞系，并研究了从TSD iPS细胞诱导出神经干细胞(NSC)和神经前体细胞(NPC)的条件。此外，我们还开发了新的荧光探针，包括Rhodol beta-GlcNAc作为细胞内Hex活性的人工底物，以及酸性pH激活的荧光Rh-PM用于体内成像，以检测重组Hex的溶酶体分布。

We studied on intracellular traffic abnormality in lysosomal beta-hexosaminidase (Hex) deficiencies including Tay-Sachs disease (TSD) and Sandhoff diseases (SD) to develop novel enzyme replacement therapy based on techniques to promote delivery of recombinant Hex to lysosomes when extracellularly administrated. We developed modified Hex with high content of mannose 6-phospahte (M6P) residues, which were efficiently incorporated by disease-model cultured cells via cell surface M6P receptors. In addition, we established an iPS cell line derived from a TSD patient, and examined conditions under which neural cells including neural stem cells (NSC) and neural progenitor cells (NPC) could be induced from the TSD iPS cells. Furthermore, we developed novel fluorescent probes including Rhodol beta-GlcNAc as an artificial substrate for intracellular Hex activity and acidic pH-activatable fluorescent Rh-PM for in vivo imaging to detect lysosomal distribution of recombinant Hex.