Therapeutic Management of Lineage- and Differentiation-state Plasticity

摘要-项目1 肿瘤内的异质性是乳腺癌治疗抵抗的一个主要原因。研究表明， 证明了异质肿瘤中的一部分癌细胞可以逃脱治疗诱导的死亡 由于先天或适应性抵抗，导致复发、疾病进展和患者生存不良。 三阴性乳腺癌是一种侵袭性疾病，具有高度的肿瘤内异质性。 和糟糕的病人结局。在初步实验中，我们确定了原发肿瘤细胞的亚群。 以及在基底样的TNBC细胞系中也是如此，这些细胞系的特征是腔，基， 和间充质分化状态标志物。我们观察到不同类别的靶向 治疗药物有能力消除或丰富这些区域内特定的分化状态亚群 株系，引导不同种类的癌细胞群体朝着更加同质性的方向发展。重要的是，我们确定了 以主调节器预测的任一途径依赖为靶点的协同组合治疗 对残留细胞或表观遗传调节因子的分析，发现有助于细胞向抗性状态的转变。 因此，这个项目的总体目标是了解细胞对治疗反应的内在调节。 表型不同的TNBC，以便开发目标策略，以杀死所有共存的 亚群。我们关注的是表型异质性，因为这可以代表遗传和 表观遗传因素，我们将利用临床相关的治疗方法来驱动异质性 种群趋向同质化。我们假设一种系统生物学的方法来测量和 对响应表型状态变化潜在的功能通路进行计算模拟 状态聚集疗法将揭示共同的逃逸途径和细胞可塑性的调节， 这将使我们能够预测有效的联合治疗策略，消除所有癌症 亚群。我们将通过(1)检查和计算建模表型来解决这一假设 多个遗传多样性、异质性的TNBC细胞系对靶向反应的状态变化 使用高含量成像和单细胞表达分析诱导同质性的治疗方法，(2) 确定克隆扩张或分化状态可塑性是否驱动动态表型变化 在有针对性的治疗和模拟这些转变背后的分子网络变化之后，以及(3) 确定状态转换背后的表观遗传调控并开发组合策略 克服异种TNBC细胞在体内外的治疗耐药。总而言之，这些目标 支持我们的目标，即测量和模拟细胞对临床相关靶向治疗的内在反应，并 预测能更有效地控制异种TNBC的协同药物组合。这一点的整合 与M2CH-CSBC中的项目2和3合作将使我们能够结合这些内在因素的外部调节 并反复改进对这一毁灭性疾病的控制策略。 好了！

ABSTRACT – Project 1 Intratumoral heterogeneity is a major cause of therapeutic resistance in breast cancer. Studies have demonstrated that a subset of cancer cells within a heterogeneous tumor can escape therapy-induced death due to innate or adaptive resistance, resulting in recurrence, disease progression, and poor patient survival. Triple negative breast cancer (TNBC) is an aggressive disease characterized by high intratumor heterogeneity and poor patient outcome. In preliminary experiments, we identified subpopulations of tumor cells in primary TNBC as well as in basal-like TNBC cell lines that are characterized by differential expression of luminal, basal, and mesenchymal differentiation-state markers. We have observed that distinct classes of targeted therapeutics have the capacity to eliminate or enrich specific differentiation-state subpopulations within these lines, steering heterogeneous cancer cell populations toward increased homogeneity. Importantly, we identified synergistic combinatorial treatments that targeted either pathway dependencies predicted by master regulator analysis of residual cells or epigenetic regulators found to contribute to a cell's transition to a resistant state. The overall goal of this project, therefore, is to understand cell intrinsic regulation of therapeutic response in phenotypically heterogeneous TNBC in order to develop targeting strategies to kill all co-existing subpopulations. We focus on phenotypic heterogeneity, as this can represent the combination of genetic and epigenetic factors, and we will take advantage of clinically relevant therapeutics that drive heterogeneous populations toward homogeneity. We hypothesize that a systems biology approach of measuring and computationally modeling the functional pathways underlying phenotypic state changes in response to state-aggregating therapeutics will reveal common escape routes and regulators of cell plasticity, which will allow us to predict effective combinatorial therapeutic strategies that eliminate all cancer subpopulations. We will address this hypothesis by (1) examining and computationally modeling phenotype state changes in multiple genetically diverse, heterogeneous TNBC cell lines in response to targeted therapeutics that induce homogeneity using high-content imaging and single cell expression analysis, (2) determining whether clonal expansion or differentiation state plasticity drives the dynamic phenotype changes following targeted therapy and modeling the molecular network changes that underlie these transitions, and (3) determining epigenetic regulation underlying state transitions and developing combinatorial strategies that overcome therapeutic resistance in heterogeneous TNBC cells in vitro and in vivo. Together, these aims support our goal to measure and model cell intrinsic responses to clinically relevant targeted therapeutics and to predict synergistic drug combinations that more effectively control heterogeneous TNBC. Integration of this work with Projects 2 and 3 in the M2CH-CSBC will allow us to incorporate extrinsic regulators of these intrinsic mechanisms and to iteratively refine control strategies for this devastating disease. !