Defining the genetic and immune landscape of precursor myeloma progression with single-cell sequencing

Multiple Myeloma is universally preceded by a pre-malignant state, however, the genetic and immune mechanisms associated with transformation are incompletely understood thus hindering development of preventative treatment strategies. Using single-cell RNA sequencing, we profiled the transcriptomes of tumor and immune microenvironment cells derived from transgenic mice whose disease spanned the myeloma progression spectrum. This work in turn provides unique and previously unappreciated insight into the biology of precursor disease progression. Overall design: BM were extracted from hind-leg bones of VkMYC/non-transgenic control mice, dissociated into a single cell suspension using mechanical disruption (syringe) and RBC's lysed. Single-cell RNA-sequencing libraries were then generated using the 10x Genomics Chromium Single Cell 3' Solution V2 protocol

多发性骨髓瘤（Multiple Myeloma）的发生均以癌前病变状态为前驱阶段，但其恶性转化相关的遗传与免疫机制尚未完全阐明，这一现状阻碍了预防性治疗策略的开发。本研究采用单细胞RNA测序技术，对疾病状态覆盖骨髓瘤进展全谱系的转基因小鼠来源的肿瘤细胞与免疫微环境细胞的转录组进行了分析。该研究进而为癌前病变进展的生物学机制提供了独特且此前未被关注的全新见解。实验设计概述：从VkMYC转基因小鼠与非转基因对照小鼠的后腿骨骼中提取骨髓（BM），通过机械破碎（注射器研磨）解离为单细胞悬液，并裂解红细胞。随后采用10x Genomics Chromium Single Cell 3' Solution V2实验流程构建单细胞RNA测序文库。