The Study of Jag1-Notch in the Extravasation of Triple Negative Breast Cancer Cells in Metastasis.

Jag1-Notch 在三阴性乳腺癌细胞转移中外渗的研究。

• 批准号: 10314566
• 负责人: Benjamin Gordon
• 金额: $ 5.1万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2025-03-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10314566
• 关键词: Study; Jag1; Notch; Extravasation; Triple

Project Summary Triple negative breast cancer (TNBC) makes up to 15-20% of breast cancer diagnoses. While surgery and adjuvant chemotherapy are effective treatment options for early-stage disease, there are very few therapeutic options for advanced metastatic TNBC, and the 11% relative five-year survival rate highlights an urgent need to discover actionable targets. In the hematogenous metastatic cascade, a few aggressive tumor cells are capable of crossing the endothelial barrier at least twice: when entering systemic circulation (intravasation) and then again when exiting circulation to colonize distant organs (extravasation). High expression of JAGGED-1 (JAG1), a Notch ligand, is strongly associated with TNBC metastasis and consequent mortality. Our preliminary work in static culture suggests that JAG1 enhances TNBC binding to the endothelium and transendothelial migration (TEM), thereby promoting dissemination of tumor cells through vascular beds. We propose to study two distinct mechanisms for JAG1 in the metastatic cascade. In Aim 1, we will investigate how tumor JAG1 signals in trans to the endothelium to prime the vascular barrier for invasion. We will examine extravasation of our recently generated TNBC CRISPR JAG1 knockout clones and control cells using a microfluidics system that faithfully models capillary fluid shear forces and permits visualization of multiple critical steps of extravasation in vitro. We will also test the role of JAG1 in vivo by examining the rate of lung capillary extravasation and pulmonary seeding following tail vein injection of JAG1 knockout and control TNBC cells. In Aim 2, we will define the tumor cell- intrinsic transcriptional program regulated by JAG1. Our preliminary RNA sequencing data suggest that novel JAG1 targets promote cell surface interactions distinct from Notch targets. We will determine the metastatic importance of key JAG1 targets by restoring expression and testing for TEM phenotypes. In the training plan of the fellowship, I highlight an integrated scientific, clinical, and educational blueprint to enhance my personal research and doctoring skills as an aspiring physician-scientist in oncology.

项目概要 三阴性乳腺癌 (TNBC) 占乳腺癌诊断的 15-20%。当手术和 辅助化疗是早期疾病的有效治疗选择，但治疗方法很少 晚期转移性 TNBC 的选择，11% 的相对五年生存率凸显了迫切需要 发现可操作的目标。在血行转移级联中，一些侵袭性肿瘤细胞能够 穿过内皮屏障至少两次：进入体循环（内渗）时，然后 当退出循环以定植远处器官时再次发生（外渗）。 JAGGED-1 (JAG1) 高表达， Notch 配体，与 TNBC 转移和随后的死亡率密切相关。我们的前期工作 静态培养表明 JAG1 增强 TNBC 与内皮的结合和跨内皮迁移 （TEM），从而促进肿瘤细胞通过血管床传播。我们建议研究两种不同的 JAG1 在转移级联中的机制。在目标 1 中，我们将研究肿瘤 JAG1 如何在反式中发出信号 到内皮细胞，为血管屏障的侵袭做好准备。我们将检查最近的外渗情况 使用微流体系统生成 TNBC CRISPR JAG1 敲除克隆和对照细胞，忠实地 模拟毛细管流体剪切力并允许体外外渗的多个关键步骤的可视化。我们 还将通过检查肺毛细血管外渗率和肺播种率来测试 JAG1 在体内的作用 尾静脉注射 JAG1 敲除细胞和对照 TNBC 细胞后。在目标 2 中，我们将定义肿瘤细胞—— 由 JAG1 调节的内在转录程序。我们的初步 RNA 测序数据表明，新的 JAG1 靶标促进细胞表面相互作用，与 Notch 靶标不同。我们将确定转移性 通过恢复表达和测试 TEM 表型来了解关键 JAG1 靶标的重要性。在培训计划中 在奖学金中，我强调了一个综合的科学、临床和教育蓝图，以增强我的个人能力 作为一名有抱负的肿瘤学医师科学家的研究和医生技能。