GBA1 regulates the CLEAR network through Calcineurin and TFEB in cellular models of Parkinson´s disease

The GBA1 gene encodes the lysosomal enzyme acid-β-glucocerebrosidase (GCase). GBA1 mutations cause the lysosomal storage disorder Gaucher disease (GD) and are a genetic risk factor for Parkinson´s disease (PD). Many GBA1 mutations cause aberrant GCase folding and processing but how these impinge on PD pathogenesis remains unclear. We addressed GCase functions in human dopaminergic (DA) neurons derived from engineered GBA1-deficient (GBA1-/-) embryonic stem cells (hESCs) and GBA1N370S PD patient-derived induced pluripotent cells (hiPSCs). GBA1-mutant DA neurons displayed aberrant morphologies and gene expression that were rescued by recombinant GCase treatment. GBA1-/- neurons have hyperactive Calcineurin (CaN) and nuclear localization of the Transcription Factor EB (TFEB). Expression of TFEB targets and lysosomal CLEAR network components were increased in GBA1-/- DA neurons and rescued by GCase replacement and CaN inhibition. Our findings reveal a link between increased CaN activity and loss of GCase, providing a mechanism for the disturbed lysosomal/autophagy pathways in GBA1-associated PD. The experiment follows a two-factor design: two genotypes (GBA+/+ and GBA-/-) and three treatments (vehicle, 1 μg/ml rGCase, and 10 μg/ml rGCase). Two biological replicates are available for each sample group. All together, 12 samples were available and profiled.

GBA1基因编码溶酶体酶酸性β葡糖脑苷脂酶（acid-β-glucocerebrosidase，简称GCase）。GBA1突变可引发溶酶体贮积病戈谢病（Gaucher disease，简称GD），同时也是帕金森病（Parkinson's disease，简称PD）的遗传风险因素。诸多GBA1突变会导致GCase的折叠与加工异常，但此类异常如何影响帕金森病的发病机制仍不明确。本研究针对源自工程化GBA1缺陷型（GBA1-/-）胚胎干细胞（hESCs）以及GBA1N370S型帕金森病患者诱导多能干细胞（hiPSCs）的人多巴胺能（DA）神经元，探究了GCase的功能。携带GBA1突变的DA神经元呈现出异常的形态与基因表达特征，该异常可通过重组GCase处理得到挽救。GBA1-/-神经元的钙调磷酸酶（Calcineurin，简称CaN）活性过高，且转录因子EB（TFEB）发生异常核定位。GBA1-/- DA神经元中，TFEB靶基因与溶酶体CLEAR网络组分的表达水平上调，且该上调可通过GCase替代疗法与CaN抑制得到逆转。本研究的发现揭示了CaN活性上调与GCase功能缺失之间的关联，为GBA1相关帕金森病中紊乱的溶酶体/自噬通路提供了潜在分子机制。本实验采用双因素设计：两种基因型（GBA+/+与GBA-/-）以及三种处理方式（空白对照、1 μg/ml 重组GCase（rGCase）、10 μg/ml 重组GCase）。每个样本组设置两个生物学重复。总计共获得12份样本并完成了表征分析。