美国哈佛大学尹鹏研究组开发出通过循环延伸放大信号实现高灵敏度单细胞质量细胞术。该研究于2024年7月29日在线发表于国际一流学术期刊《自然—生物技术》。

研究人员提出了一种信号放大技术，称为循环扩展放大（ACE），它结合了基于热循环的DNA原位连接和基于3-cyanovinylcarbazole phosphoramidite的DNA交联，以同时在超过30种蛋白表位上实现信号放大。研究人员展示了ACE在低丰度蛋白定量中的实用性，利用悬浮质谱细胞术来表征上皮到间质转化以及间质到上皮转化过程中的分子重编程。

研究人员展示了ACE在定量人T淋巴细胞信号网络反应动态方面的能力。研究人员进一步展示了ACE在基于成像质谱细胞术的多参数组织成像中的应用，以识别组织区室并描述与多囊肾组织中病理状态相关的空间方面。

据悉，质谱细胞术利用金属同位素标记抗体标记感兴趣的靶标，这使得能够在数百万个单细胞中同时测量约50种蛋白质或蛋白质修饰，但其灵敏度有限。

附：英文原文

Title: Signal amplification by cyclic extension enables high-sensitivity single-cell mass cytometry

Author: Lun, Xiao-Kang, Sheng, Kuanwei, Yu, Xueyang, Lam, Ching Yeung, Gowri, Gokul, Serrata, Matthew, Zhai, Yunhao, Su, Hanquan, Luan, Jingyi, Kim, Youngeun, Ingber, Donald E., Jackson, Hartland W., Yaffe, Michael B., Yin, Peng

Issue&Volume: 2024-07-29

Abstract: Mass cytometry uses metal-isotope-tagged antibodies to label targets of interest, which enables simultaneous measurements of ~50 proteins or protein modifications in millions of single cells, but its sensitivity is limited. Here, we present a signal amplification technology, termed Amplification by Cyclic Extension (ACE), implementing thermal-cycling-based DNA in situ concatenation in combination with 3-cyanovinylcarbazole phosphoramidite-based DNA crosslinking to enable signal amplification simultaneously on >30 protein epitopes. We demonstrate the utility of ACE in low-abundance protein quantification with suspension mass cytometry to characterize molecular reprogramming during the epithelial-to-mesenchymal transition as well as the mesenchymal-to-epithelial transition. We show the capability of ACE to quantify the dynamics of signaling network responses in human T lymphocytes. We further present the application of ACE in imaging mass cytometry-based multiparametric tissue imaging to identify tissue compartments and profile spatial aspects related to pathological states in polycystic kidney tissues.

DOI: 10.1038/s41587-024-02316-x

Source: https://www.nature.com/articles/s41587-024-02316-x