State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center (ChemBIC), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
Department of Biochemistry, Albert Einstein College of Medicine of Yeshiva University, 1301 Morris Park Avenue, Bronx, New York 10461, United States.
J Am Chem Soc. 2023 Jun 14;145(23):12701-12716. doi: 10.1021/jacs.3c02602. Epub 2023 Jun 5.
Despite the rich information about the physiological state of a cell encoded in the dynamic changes of cell-surface glycans, chemical methods to capture specific glycan epitopes at the single-cell level are quite limited. Here, we report a chemoenzymatic method for the single-cell detection of N-acetyllactosamine (LacNAc) by labeling LacNAc with a specific DNA barcode. The chemoenzymatic labeling does not alter the transcriptional status of immune cells and is compatible with multiple scRNA-seq platforms. Integrated analysis of LacNAc and the transcriptome of T cells at the single-cell level reveals that the amount of cell-surface LacNAc is significantly upregulated in activated CD8 T cells but maintained at basal levels in resting CD8 T cells (i.e., naive and central memory T cells). Further analysis confirms that LacNAc levels are positively correlated with the glycolytic activity of CD8 T cells during differentiation. Taken together, our study demonstrates the feasibility of the chemoenzymatic detection of cell-surface glycan in single-cell RNA sequencing-based multiomics with TCR sequence and cell-surface epitope information (i.e., scTCR and CITE-seq), and provides a new way to characterize the biological role of glycan in diverse physiological states.
尽管细胞表面糖链的动态变化中蕴含着丰富的细胞生理状态信息，但在单细胞水平上捕捉特定糖基表位的化学方法相当有限。在这里，我们报告了一种通过用特定的 DNA 条码标记乳糖胺 (LacNAc) 来实现单细胞检测 N-乙酰乳糖胺 (LacNAc) 的化学生物酶方法。该化学生物酶标记法不会改变免疫细胞的转录状态，并且与多个 scRNA-seq 平台兼容。在单细胞水平上对 LacNAc 和 T 细胞的转录组进行综合分析表明，在激活的 CD8 T 细胞中，细胞表面 LacNAc 的含量显著上调，但在静止的 CD8 T 细胞（即幼稚和中央记忆 T 细胞）中保持基础水平。进一步的分析证实，LacNAc 水平与 CD8 T 细胞在分化过程中的糖酵解活性呈正相关。综上所述，我们的研究证明了在基于 TCR 序列和细胞表面表位信息（即 scTCR 和 CITE-seq）的单细胞 RNA 测序的多组学中进行细胞表面聚糖的化学生物酶检测的可行性，并为在不同生理状态下糖在生物中的作用提供了一种新的表征方法。
