多发性骨髓瘤(MM) 是一种高度难治的血液癌症。针对性免疫疗法对MM显示出了希望，但仍受到确定特定但广泛代表性肿瘤标记物的挑战的限制。我们使用单细胞转录组学方法，对41名MM患者的53个骨髓穿刺样本进行了疗法靶点发现的无偏和高通量的管道分析，得到了38个编码细胞表面蛋白和15个编码细胞内蛋白的MM标记基因。其中，突出了20个目前未在临床研究中的候选基因，其中11个之前未被表征为治疗靶点。使用MM细胞系和患者骨髓的批量RNA测序、流式细胞术和蛋白质质谱分析进行交叉验证，表明了数据类型之间的高度一致性。使用批量RNA测序进行独立验证重申了顶级候选基因，进一步证实了单细胞转录组学方法能够准确捕捉到标记表达，尽管样本量或测序深度有限。使用转录组和免疫成像方法进一步研究了靶标的动态和异质性。总之，这项研究提出了一种强大且广泛适用的策略，以确定肿瘤标记物，从而更好地指导靶向癌症治疗的发展。

Multiple myeloma (MM) is a highly refractory hematological cancer. Targeted immunotherapy has shown promise in MM but remains hindered by the challenge of identifying specific yet broadly representative tumor markers. We analyzed 53 bone marrow (BM) aspirates from 41 MM patients using an unbiased, high-throughput pipeline for therapeutic target discovery via single cell transcriptomic profiling, yielding 38 MM marker genes encoding cell-surface proteins and 15 encoding intracellular proteins. Of these, 20 candidate genes were highlighted that are not yet under clinical study, 11 of which are previously uncharacterized as therapeutic targets. The findings were cross-validated using bulk RNA-sequencing, flow cytometry, and proteomic mass spectrometry of MM cell lines and patient BM, demonstrating high overall concordance across data types. Independent discovery using bulk RNA-sequencing reiterated top candidates, further affirming the ability of single cell transcriptomics to accurately capture marker expression despite limitations in sample size or sequencing depth. Target dynamics and heterogeneity were further examined using both transcriptomic and immuno-imaging methods. In summary, this study presents a robust and broadly applicable strategy for identifying tumor markers to better inform the development of targeted cancer therapy.