Integrators of Metastatic Potential

摘要 NCI 精准医学计划的一个关键目标是开发针对癌症的新工具 针对疾病状态和转移风险的治疗——转移性疾病高风险患者将接受 积极的、经常进行分子靶向治疗，而那些转移性疾病风险较低的人会 采用适当的局部疗法进行治疗，避免治疗的毒副作用，同时保持高水平 治愈的可能性。阻碍精准医学转移治疗的主要挑战之一 对赋予小细胞高转移潜力的信号分子和途径的了解有限 较大的异质肿瘤内的癌细胞子集。因此，转移性肿瘤的分子成像 “潜力”是一个尚未克服的挑战。 设计能够检测和测量单个活癌细胞转移“潜力”的生物传感器， 我们对泛癌磷酸蛋白质组进行了全面分析，以寻找肌动蛋白重塑剂 细胞迁移所需的，在癌症中富集，但在正常细胞中排除。只有一种磷蛋白 出现了酪氨酸磷酸化的 CCDC88A (GIV/Girdin)，这是一种跨多种细胞的真正的转移相关蛋白 实体瘤。我们设计的多模块生物传感器部分源自 GIV，并且因为 GIV 通过多种致癌受体整合促转移信号，我们将它们命名为“转移整合器” 潜力（IMP）'。假设使用 IMP 对 GIV 激活进行单细胞成像，而不是简单地 GIV 表达水平，将检测必须消除以防止转移起始细胞的子集 转移性疾病。初步实验表明 IMP 捕获了转移性肿瘤的异质性 在稳态下原发性肺和乳腺肿瘤中的潜力，检测到那些已经获得的少数细胞 最高的转移潜力并追踪其在转移过程中的富集。这些发现提供了证据—— IMP 可以测量敏感肿瘤细胞转移潜能的多样性和可塑性 和公正的方式。展望未来，IMP 将被优化和验证，以用作分子成像工具 通过 3 个目标来确定转移起始细胞：1）对单个乳腺癌细胞的转移潜力进行成像； 2）图像 GIV 激活作为转移起始细胞 (MIC) 的标志； 3) 患者的影像转移潜力- 衍生的异种移植物。为了实现这些目标，这个多 PD/PI 提案利用了非 两个 PI 和两个动物模型系统（斑马鱼和小鼠）的重叠专业知识，新颖的深入生物学 信号通路、体内成像前沿技术。 成功检测转移起始细胞将是癌细胞生物学的革命性进步。 它将为个性化筛查平台的开发铺平道路，以评估抗病毒药物的有效性。 癌症药物能够杀死任何特定肿瘤内的高致瘤性细胞，并最终使临床成为可能 实施精准癌症治疗，以改善患者的治疗和生活质量。

ABSTRACT One key goal of the NCI Precision Medicine Initiative focuses on development of new tools to tailor cancer therapy to disease status and risk of metastasis-- Patients at high risk for metastatic disease would receive aggressive, frequently molecularly-targeted therapy, whereas those with low risk for metastatic disease would be treated with appropriate local therapies, sparing them toxic side effects of therapy while maintaining high likelihood for cure. One of the main challenges preventing implementation of precision medicine for metastasis is limited understanding of signaling molecules and pathways that confer high metastatic potential to a small subset of cancer cells within a larger, heterogeneous tumor. Consequently, molecular imaging of metastatic ‘potential’ is an unvanquished challenge. To engineer biosensors that can detect and measure metastatic 'potential' of single living cancer cells, we carried out a comprehensive analysis of the pan-cancer phosphoproteome to search for actin-remodelers required for cell migration, that are enriched in cancers, but excluded in normal cells. Only one phosphoprotein emerged, tyr-phosphorylated CCDC88A (GIV/Girdin), a bona-fide metastasis-related protein across a variety of solid tumors. We designed multi-modular biosensors that are partly derived from GIV, and because GIV integrates pro-metastatic signaling by multiple oncogenic receptors, we named them ‘Integrator-of-Metastatic- Potential (IMP)'. It is hypothesized that single cell imaging of GIV activation using IMPs, rather than simply levels of GIV expression, will detect the subset of metastasis-initiating cells that must be eliminated to prevent metastatic disease. Preliminary experiments demonstrate that IMPs captured the heterogeneity of metastatic potential within primary lung and breast tumors at steady-state, detected those few cells which have acquired the highest metastatic potential and tracked their enrichment during metastasis. These findings provide proof- of-concept that IMPs can measure the diversity and plasticity of metastatic potential of tumor cells in a sensitive and unbiased way. Going forward, IMPs will be optimized and validated for use as tools for molecular imaging of metastasis-initiating cells via 3 goals: 1) Image metastatic potential of single breast cancer cells; 2) Image GIV activation as a marker of Metastasis Initiating Cells (MICs); and 3) Image metastatic potential in patient- derived xenografts. To accomplish these goals, this multi PD/PI proposal capitalizes on synergy of non- overlapping expertise of two PIs and two animal model systems (zebrafish and mice), in-depth biology of a novel signaling pathway, and cutting-edge technology of in vivo imaging. Success in detecting metastasis-initiating cells will be a transformative advance for cancer cell biology. It will pave the path for the development of personalized screening platforms to assess the effectiveness of anti- cancer drugs in killing the highly tumorigenic cells within any given tumor, and ultimately enable clinical implementation of precision cancer therapy to improve treatment and quality of life for patients.