An altered secretome is an early marker of the pathogenesis of CLN6 Batten disease.
An altered secretome is an early marker of the pathogenesis of CLN6 Batten disease.
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分泌蛋白组的改变是 CLN6 Batten 病发病机制的早期标志。
DOI：
10.1111/jnc.15285
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发表时间：
2021
影响因子：
4.7
通讯作者：
Best HL
中科院分区：
文献类型：
--
作者：
Best HL
神经元蜡样质脂褐质沉积症（NCL）是一组遗传性儿童神经退行性疾病。除了自身荧光储存物质在溶酶体中的积累外，NCL的主要特征是区域特异性神经炎症，可以预测神经元损失。这些表型表明细胞外环境的改变，使分泌组成为一个重要的研究领域。本研究调查了NCL的CLN 6（蜡样脂褐质沉积症神经元蛋白6）变体中的分泌组。为了研究CLN 6分泌组，我们将从Cln 6 ±小鼠的Cln 6 ncl分离的神经元和胶质细胞共培养，并利用质谱法比较条件培养基的蛋白质组分。通过蛋白质印迹和酶活性测定进一步研究了组织蛋白酶中观察到的显著变化。病毒介导的基因治疗用于尝试和拯救野生型表型并恢复分泌组-在体外共培养物和体内小鼠血浆中。在Cln 6 nclf细胞中，蛋白质组学显示分解代谢和细胞骨架相关蛋白质显著增加，揭示了NCL与其他神经退行性疾病的致病特征之间的新相似性。这些变化部分通过基因治疗干预得到纠正，表明这些蛋白质可作为体外生物标志物的候选物。重要的是，这些体外变化显示出体内翻译的前景，在基因治疗后的共培养物和Cln 6 nclf血浆样品中，组织蛋白酶L（CTSL）活性降低。这项工作表明分泌组在CLN 6发病机制中起作用，并突出了其作为体外模型的潜在用途。蛋白质组学的变化提供了一系列候选生物标志物，用于监测疾病和评估未来研究中的潜在治疗方法。
Neuronal ceroid lipofuscinoses (NCLs) are a group of inherited childhood neurodegenerative disorders. In addition to the accumulation of auto‐fluorescent storage material in lysosomes, NCLs are largely characterised by region‐specific neuroinflammation that can predict neuron loss. These phenotypes suggest alterations in the extracellular environment—making the secretome an area of significant interest. This study investigated the secretome in the CLN6 (ceroid‐lipofuscinosis neuronal protein 6) variant of NCL. To investigate the CLN6 secretome, we co‐cultured neurons and glia isolated fromCln6nclfor Cln6±mice, and utilised mass spectrometry to compare protein constituents of conditioned media. The significant changes noted in cathepsin enzymes, were investigated further via western blotting and enzyme activity assays. Viral‐mediated gene therapy was used to try and rescue the wild‐type phenotype and restore the secretome—both in vitro in co‐cultures and in vivo in mouse plasma. InCln6nclfcells, proteomics revealed a marked increase in catabolic and cytoskeletal‐associated proteins—revealing new similarities between the pathogenic signatures of NCLs with other neurodegenerative disorders. These changes were, in part, corrected by gene therapy intervention, suggesting these proteins as candidate in vitro biomarkers. Importantly, these in vitro changes show promise for in vivo translation, with Cathepsin L (CTSL) activity reduced in both co‐cultures andCln6nclfplasma samples post gene‐therapy. This work suggests the secretome plays a role in CLN6 pathogenesis and highlights its potential use as an in vitro model. Proteomic changes present a list of candidate biomarkers for monitoring disease and assessing potential therapeutics in future studies.