Single-cell RNA-Sequencing of neurons labeled by retrograde FastBlue tracing (BarcodeSeq)

Most cancers including pancreatic ductal adenocarcinoma (PDAC) are infiltrated by PNS neurons participating in their complex tumor microenvironment. However, their cell bodies and nuclei are located in the para- and pre-vertebral PNS ganglia located far from the tumor mass itself. Thus, molecular information on healthy organ- vs. cancer-infiltrating neurons is currently lacking in any sequencing dataset of healthy or tumor tissue. To specifically identify and molecularly characterize the identity and transcriptomes of PDAC-infiltrating neurons at single cell resolution, we developed "Trace-n-seq". This method is based on retrograde tracing of axons from target tissues to their respective ganglia, followed by individual FACS-isolation and transcriptomic analysis. We characterized >2000 sympathetic and sensory neurons that infiltrate PDAC, healthy pancreas, or other abdominal organs.包括胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）在内的多数癌症，其复杂肿瘤微环境中均存在外周神经系统（Peripheral Nervous System, PNS）神经元浸润。然而此类神经元的胞体与细胞核均位于远离肿瘤本体的椎旁及椎前外周神经节内。因此，当前的健康组织或肿瘤组织测序数据集中，仍缺乏健康器官来源与癌症浸润神经元的分子特征对比信息。为在单细胞分辨率下精准鉴定胰腺导管腺癌浸润神经元的身份特征并解析其转录组，我们开发了"Trace-n-seq"技术。该技术基于逆向示踪靶组织轴突至对应神经节的原理，后续辅以单个细胞的荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS）分离与转录组分析。我们已对浸润胰腺导管腺癌、健康胰腺或其他腹部器官的逾2000个交感与感觉神经元进行了特征表征。