Circulating tumor cells (CTCs) that undergo epithelial-to-mesenchymal transition (EMT) can provide valuable information regarding metastasis and potential therapies. However, current studies on the EMT overlook alternative splicing. Here, we used single-cell full-length transcriptome data and mRNA sequencing of CTCs to identify stage-specific alternative splicing of partial EMT and mesenchymal states during pancreatic cancer metastasis. We classified definitive tumor and normal epithelial cells via genetic aberrations and demonstrated dynamic changes in the epithelial-mesenchymal continuum in both epithelial cancer cells and CTCs. We provide the landscape of alternative splicing in CTCs at different stages of EMT, uncovering cell-type-specific splicing patterns and splicing events in cell surface proteins suitable for therapies. We show that governs cell fate through alternative splicing independently of changes in gene expression and affects the splicing pattern during EMT. We found a high frequency of events that contained multiple premature termination codons and were enriched with C and G nucleotides in close proximity, which influence the likelihood of stop codon readthrough and expand the range of potential therapeutic targets. Our study provides insights into the EMT transcriptome’s dynamic changes and identifies potential diagnostic and therapeutic targets in pancreatic cancer.

中文翻译：

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胰腺循环肿瘤上皮-间质转化中选择性剪接的功能要求

经历上皮间质转化（EMT）的循环肿瘤细胞（CTC）可以提供有关转移和潜在疗法的有价值的信息。然而，目前关于 EMT 的研究忽视了选择性剪接。在这里，我们使用单细胞全长转录组数据和 CTC 的 mRNA 测序来识别胰腺癌转移过程中部分 EMT 和间质状态的阶段特异性选择性剪接。我们通过遗传畸变对确定的肿瘤和正常上皮细胞进行分类，并证明了上皮癌细胞和 CTC 中上皮间质连续体的动态变化。我们提供了 EMT 不同阶段 CTC 中选择性剪接的情况，揭示了适合治疗的细胞类型特异性剪接模式和细胞表面蛋白中的剪接事件。我们表明，它通过独立于基因表达变化的选择性剪接来控制细胞命运，并影响 EMT 期间的剪接模式。我们发现包含多个提前终止密码子的事件频率很高，并且富含接近的 C 和 G 核苷酸，这影响了终止密码子通读的可能性并扩大了潜在治疗靶点的范围。我们的研究提供了对 EMT 转录组动态变化的见解，并确定了胰腺癌的潜在诊断和治疗靶点。