Targeting plasticity in lung cancer

摘要 肺腺癌(LUAD)是非小细胞肺癌最常见的亚型，导致约55,000人死亡 美国每年都有死亡病例。尽管最近在LUAD治疗方面取得了进展，但该病仍然非常严重 很难对付。因此，对于LUAD的新的和有效的治疗策略存在着严重的未得到满足的临床需求 病人。癌细胞的可塑性-分化和适应细胞的能力-外部压力-驱动肿瘤 进展，是LUAD治疗失败的主要原因。因此，LUAD的靶向可塑性是一个很有前途的研究方向 治疗理念。实现靶向癌细胞可塑性的治疗潜力需要基本的 对LUAD中促进可塑性的细胞状态以及驱动的分子机制的理解 他们。使用LUAD基因工程小鼠模型(GEMM)和单细胞mRNA测序 (scRNA-Seq)为了研究LUAD的进化，我们发现了一种高塑性细胞状态(HPCS)，它是由 处于肿瘤进化早期的LUAD细胞的一个亚群。HPCS在小鼠体内普遍存在， 体内人类LUAD不分分期和相当大的瘤内和瘤间遗传和表型 多样性。此外，HPCS基因表达特征与特别差的患者预后相关。 预期分离的HPCS细胞具有强大的分化能力(可塑性)和 化疗后HPCS明显丰富。我们的前期工作力度很大 支撑物的可塑性集中在HPCS中，它与高增长潜力和 化疗耐药。然而，HPCS对LUAD增长、治疗耐药性、 或在LUAD肿瘤内出现新的恶性细胞状态尚不清楚。同样，人们对此知之甚少。 LUAD可塑性的转录驱动因素。我们假设，HPCS对于 癌前肿瘤转化为LUAD以及LUAD生长、细胞状态转变和化疗耐药。至 解决这一假设，我们将询问HPCs在LUAD进展和治疗耐药使用谱系- 使用我们生成的新型报告系统进行消融和血统追踪，我们将与 ScRNA-seq.为了解决分子HPCS驱动因素，我们将失活或过度表达两个候选转录 LUAD GEMM和人类患者来源的异种移植(PDX)LUAD模型中的因素，其次是基因 表达和染色质可及性分析。我们建议的研究将使我们能够建立HPCS，一个 以前未知的细胞状态，是根除LUAD可塑性的关键。这将导致一种新的治疗方法 范式，激发跨实体肿瘤的高可塑性细胞状态的靶向。此外，我们的工作将 为体内肿瘤细胞状态异质性的原位研究提供了一个新的平台。

SUMMARY Lung adenocarcinoma (LUAD), the most common subtype of non-small cell lung cancer, results in ~55,000 deaths in the US every year. Despite the recent advancements in LUAD treatment, the disease remains highly intractable. Thus, a critical unmet clinical need exists for novel and effective therapeutic strategies for LUAD patients. Cancer cell plasticity – the capacity to differentiate and adapt to cell-extrinsic pressure – drives tumor progression and is a major cause of treatment failure in LUAD. Thus, targeting plasticity in LUAD is a promising therapeutic concept. Realizing the therapeutic potential of targeting cancer cell plasticity requires fundamental understanding of the cell states that promote plasticity in LUAD as well as the molecular mechanisms that drive them. Using a genetically engineered mouse model (GEMM) of LUAD and single-cell mRNA sequencing (scRNA-Seq) to investigate LUAD evolution we identified a high-plasticity cell state (HPCS) that is acquired by a subset of LUAD cells in early stages of tumor evolution. The HPCS was ubiquitously maintained in mouse and human LUAD in vivo irrespective of stage and considerable intra- and inter-tumoral genetic and phenotypic diversity. Further, the HPCS gene expression signature correlated with particularly poor patient outcomes. Prospectively isolated HPCS cells were endowed with robust capacity for differentiation (plasticity) and proliferation, and the HPCS was strongly enriched following chemotherapy. Our preliminary work strongly supports plasticity is concentrated in the HPCS and that it is associated with high growth potential and chemoresistance. However, the contribution or essentiality of the HPCS for LUAD growth, treatment resistance, or emergence of new malignant cell states within LUAD tumors is not known. Similarly, little is known of the transcriptional drivers of LUAD plasticity. We hypothesize that the HPCS is essential for progression of premalignant neoplasias to LUAD as well as for LUAD growth, cell state transitions, and chemoresistance. To address this hypothesis, we will interrogate HPCS in LUAD progression and treatment resistance using lineage- ablation and lineage-tracing using a novel reporter system that we have generated, which we will combine with scRNA-seq. To address molecular HPCS drivers, we will inactivate or overexpress two candidate transcription factors in the LUAD GEMM and in human patient-derived xenograft (PDX) LUAD models, followed by gene expression and chromatin accessibility profiling. Our proposed study will allow us to establish the HPCS, a previously unknown cell state, as key to eradicating plasticity in LUAD. This would lead to a new treatment paradigm, motivating targeting of high-plasticity cell states across solid tumors. Furthermore, our work will contribute a novel platform for the in situ investigation of cell state heterogeneity in cancers in vivo.