Discovering proteins that explain single-cell heterogeneity in fibrillar migration

项目摘要 乳腺癌的异质性令人望而生畏，即使在单个患者的肿瘤内也是如此。这个 临床结果是，针对某些癌细胞的靶向治疗作用仍然无效。 与那些已经进化出替代途径来完成疾病驱动功能的同行形成对比。 因此，迫切需要单细胞方法来发现跨越 异质癌细胞转移的互补途径。转移的早期步骤 包括癌细胞通过原发肿瘤微环境侵袭到附近的血管和 淋巴网络，它们通过这个网络传播到次级站点。乳腺癌的一种常见类型 原发肿瘤微环境的侵袭涉及癌细胞沿胶原纤维的迁移。这个 调控眼球移动的蛋白质知之甚少，以及调控眼球运动的各种途径 乳腺癌亚型实现了这种侵袭性的迁移模式，这一点还有待阐明。在 提出的工作，我们寻求开发，优化和应用单细胞迁移和蛋白质组学(ScMAP)， 一种对同一个体的迁移特性和蛋白质组进行量化的创新方法 手机。因此，scMAP能够收集直接连接细胞迁移和 在单细胞水平上存在数以千计的蛋白质。我们将应用scMAP来发现蛋白质 其丰度与一组异质性乳腺癌的纤毛转移增强有关 细胞系。用总体最小二乘回归和互信息分析这一新的数据集 将揭示与纤毛虫迁移呈定量线性和非线性关系的蛋白质 属性。这些结果将确定不同的蛋白质组，以解释不同的纤毛虫迁移 异质乳腺癌细胞的特性。识别的蛋白质为未来的功能提供了管道 在复杂的3D睫毛基质中进行测试和随后潜在的治疗应用来抑制睫毛 侵入性迁徙。此外，scMAP为蛋白质组规模的单细胞分辨率洞察打开了大门 在其他情况下进入细胞迁移，包括转移中的其他运动模式。

Project Summary The daunting heterogeneity of breast cancer is apparent even within a tumor in an individual patient. The clinical consequence is that a targeted therapeutic e ective against some cancer cells remains impotent against counterparts that have evolved alternative pathways to accomplish disease-driving functions. Thus, there is an urgent need for single-cell approaches to discover molecular targets that span the complementary pathways by which heterogeneous cancer cells metastasize. An early step in metastasis involves cancer cells invading through the primary tumor microenvironment to reach nearby vascular and lymphatic networks through which they disseminate to secondary sites. A common mode of breast cancer invasion in the primary tumor microenvironment involves cancer cell migration along collagen bers. The proteins that regulate brillar migration are poorly understood, and the diversity of pathways by which breast cancer subtypes accomplish this invasive mode of migration remains to be elucidated. In the proposed work, we seek to develop, optimize and apply single-cell migration and proteomics (scMAP), an innovative method to quantify both the migration properties and the proteome of the same individual cell. Thus, scMAP enables the collection of a novel data set that directly links cell migration and the abundance of thousands of proteins at the single-cell level. We will apply scMAP to discover proteins whose abundance is associated with enhanced brillar migration of a panel of heterogenous breast cancer cell lines. Analysis of this novel data set by total least squares regression and mutual information will reveal proteins that exhibit quantitative linear and non-linear relationships to brillar migration properties. These results will identify distinct sets of proteins that explain the disparate brillar migration properties of heterogeneous breast cancer cells. The identi ed proteins feed a pipeline for future functional testing in complex 3d brillar matrices and subsequent potential therapeutic application to curb brillar invasive migration. In addition, scMAP opens the door to proteomic-scale, single-cell resolution insights into cell migration in other contexts, including other modes of motility in metastasis.