A DNA nanomachine chemically resolves lysosomes in live cells
A DNA nanomachine chemically resolves lysosomes in live cells
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DOI：
10.1038/s41565-018-0318-5
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发表时间：
2019-02-01
影响因子：
38.3
通讯作者：
Krishnan, Yamuna
中科院分区：
文献类型：
--
作者：
Leung, KaHo;Chakraborty, Kasturi;Krishnan, Yamuna
溶酶体是多功能的亚细胞器，在质膜修复、自噬、病原体降解和营养感测中起作用。溶酶体功能障碍是阿尔茨海默病、帕金森病和罕见的溶酶体贮积病的基础，但它们对这些病理生理学的贡献尚不清楚。活体成像显示溶酶体亚群具有不同的物理特性，包括动力学，形态学或细胞定位。在这里，我们化学解析溶酶体亚群使用基于DNA的组合报告，定量图像的pH值和氯化物同时在同一个溶酶体，同时保留单溶酶体的活细胞中的信息。我们称这种技术为双离子测量或2-IM。来源于健康个体的原代皮肤成纤维细胞中的溶酶体的2-IM显示两个主要的溶酶体群体，其中一个在来源于尼曼-匹克病患者的原代细胞中不存在。当患者细胞用相关疗法处理时，第二群体重新出现。通过2-IM化学解析溶酶体可以解码溶酶体疾病的机制基础，监测疾病进展或评估治疗效果。
Lysosomes are multifunctional, subcellular organelles with roles in plasma membrane repair, autophagy, pathogen degradation and nutrient sensing. Dysfunctional lysosomes underlie Alzheimer's disease, Parkinson's disease and rare lysosomal storage diseases, but their contributions to these pathophysiologies are unclear. Live imaging has revealed lysosome subpopulations with different physical characteristics including dynamics, morphology or cellular localization. Here, we chemically resolve lysosome subpopulations using a DNA-based combination reporter that quantitatively images pH and chloride simultaneously in the same lysosome while retaining single-lysosome information in live cells. We call this technology two-ion measurement or 2-IM. 2-IM of lysosomes in primary skin fibroblasts derived from healthy individuals shows two main lysosome populations, one of which is absent in primary cells derived from patients with Niemann-Pick disease. When patient cells are treated with relevant therapeutics, the second population re-emerges. Chemically resolving lysosomes by 2-IM could enable decoding the mechanistic underpinnings of lysosomal diseases, monitoring disease progression or evaluating therapeutic efficacy.