Tumor microenvironment landscapes supporting EGFR-mutant NSCLC are modulated at the single cell interaction level by Unesbulin treatment

Lung cancer is the leading cause of cancer deaths. Lethal pulmonary adenocarcinomas (ADCs) present with frequent mutations in the Epidermal Growth Factor Receptor (EGFR). Genetically-engineered murine models of lung cancer expedited comprehension of the molecular mechanisms driving tumorigenesis and drug response. Here, we systematically analyzed the evolution of tumor heterogeneity in the context of dynamic interactions occurring with the intermingled tumor microenvironment by high resolution transcriptomics. Our effort identified vulnerable tumor-specific epithelial cells, as well as their cross-talk with niche components (endothelial cells, fibroblasts and tumor-infiltrating immune cells), whose symbiotic interface shapes tumor aggressiveness and is almost completely abolished by treatment with Unesbulin, a tubulin binding agent that reduces BMI-1 activity. Simultaneous MRI analysis demonstrated decreased tumor growth, setting the stage for future investigations into the potential of novel therapeutic strategies for EGFR-mutant ADCs. Overall design: Single cell RNA sequencing (scRNAseq) of lung tissues from EGFR-mutant model mice using InDrops-seq. The dataset includes five samples, each sequenced with distinct index sequences. Upon demultiplexing with these indexes, the dataset includes a total of 17 samples, comprising 2 untreated healthy samples, 2 healthy samples treated with a vehicle, 2 healthy samples treated with Unesbulin, 8 tumor samples treated with a vehicle, and 3 tumor samples treated with Unesbulin.

肺癌是癌症相关死亡的首要病因。致死性肺腺癌（pulmonary adenocarcinomas, ADCs）常携带表皮生长因子受体（Epidermal Growth Factor Receptor, EGFR）高频突变。基因工程构建的肺癌小鼠模型，极大推动了肿瘤发生与药物应答相关分子机制的解析。本研究通过高分辨率转录组学技术，系统分析了肿瘤微环境动态互作背景下的肿瘤异质性演化过程。研究鉴定出具有脆弱性的肿瘤特异性上皮细胞，及其与微环境组分（内皮细胞、成纤维细胞及肿瘤浸润免疫细胞）的交叉互作；二者形成的共生界面可调控肿瘤侵袭性，而使用微管结合剂Unesbulin（可降低BMI-1活性）进行治疗，几乎可完全阻断该界面的功能。同步磁共振成像（Magnetic Resonance Imaging, MRI）分析显示肿瘤生长受到显著抑制，为后续探索EGFR突变型肺腺癌的新型治疗策略奠定了基础。实验整体设计：采用InDrops-seq技术对EGFR突变模型小鼠的肺组织开展单细胞RNA测序（single cell RNA sequencing, scRNAseq）。本数据集初始包含5份样本，每份样本均使用独特的索引序列进行测序。通过上述索引序列进行解复用后，数据集最终包含17份样本，其中包括2份未处理的健康对照样本、2份赋形剂处理的健康样本、2份Unesbulin处理的健康样本、8份赋形剂处理的肿瘤样本，以及3份Unesbulin处理的肿瘤样本。