Dissecting the interplay between biomechanics and ERK signaling during tumor invasion

剖析肿瘤侵袭过程中生物力学和 ERK 信号传导之间的相互作用

• 批准号: 459686752
• 负责人: Dr. Sandra Lemke
• 金额: --
• 依托单位: Department of Chemical and Biological Engineering
• 依托单位国家: 德国
• 项目类别: WBP Fellowship
• 财政年份: 2021
• 资助国家: 德国
• 起止时间: 2020-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/459686752?language=en
• 关键词: Dissecting; interplay; between; biomechanics; ERK

The ducts of the mammary gland form an extensive branched network that allows the transport of milk. Breast epithelial tissue can also give rise to breast cancer, the most common cancer type in women. Breast cancer-related deaths typically involve the formation of metastases from tumors that invade the surrounding tissue. To block invasive metastasis, it is crucial to understand the mechanisms that drive invasion. In recent years, it has become evident that intercellular heterogeneity and signaling dynamics can profoundly impact gene expression (Davies et al. 2020) and cell behavior, such as proliferation, invasion, and collective migration (Hallou et al. 2017; Bugaj et al. 2018). Dysregulation of the extracellular-signal regulated kinase (ERK) signaling pathway has been widely implicated in breast cancer (McCain 2013), but neoadjuvant chemotherapy, which alters ERK signaling, paradoxically is associated with an increase in metastasis in some patients and animal models (Perelmuter et al. 2019). Characterization of ERK signaling dynamics at the single-cell level in invasive 3D tissues has been technically challenging. Here, we hypothesize that tissue geometry and ERK signaling are integrated via mechanical feedback to regulate invasion. To address this hypothesis, we will combine bioengineering tools with recently developed fluorescent markers, signaling reporters, optogenetic tools, and 3D cell tracking to gain valuable insights into how cells regulate their dynamic signaling behavior according to their positioning within a tissue. In Aim 1, we will create 3D bioengineered tissues with predefined geometries to determine how geometrical constraints affect the ability of mammary epithelial tissues to invade. In Aim 2, we will perform time-lapse confocal microscopy analysis using fluorescent reporters to track ERK signaling dynamics in all cells within these bioengineered tissues and define how cell shape and positioning regulate ERK signaling during invasion. We will also test whether activation of the ERK pathway is sufficient to induce or inhibit invasions by using optogenetic tools that permit spatial and temporal control. In Aim 3, we will apply recently developed optogenetic tools to induce cell contractions specifically in certain cells within 3D tissues to define the effects of mechanical forces on ERK signaling and invasion. In conclusion, this work will reveal how tissue geometry, mechanical forces, and ERK signaling dynamics interact to drive tumor invasion, and thereby define the key steps of the early breast cancer metastatic cascade, which could be interrupted therapeutically. Furthermore, this work will provide new avenues to monitor and manipulate signaling within tumor cells, which is broadly applicable to other types of cancer.

乳腺导管形成一个广泛的分支网络，允许牛奶的运输。乳腺上皮组织也会导致乳腺癌，这是女性最常见的癌症类型。乳腺癌相关死亡通常涉及肿瘤侵袭周围组织的转移形成。为了阻断侵袭性转移，了解驱动侵袭的机制至关重要。近年来，很明显，细胞间异质性和信号传导动力学可以深刻影响基因表达（Davies et al. 2020）和细胞行为，如增殖、侵袭和集体迁移（Hallou et al. 2017; Bugaj et al. 2018）。细胞外信号调节激酶（ERK）信号通路的失调广泛涉及乳腺癌（McCain 2013），但新辅助化疗改变了ERK信号通路，矛盾的是，新辅助化疗与一些患者和动物模型中的转移增加相关（Perelmuter et al. 2019）。在侵袭性3D组织中在单细胞水平上表征ERK信号传导动力学在技术上具有挑战性。在这里，我们假设，组织的几何形状和ERK信号通过机械反馈整合，以调节入侵。为了解决这一假设，我们将联合收割机生物工程工具与最近开发的荧光标记物，信号报告，光遗传学工具和3D细胞跟踪相结合，以获得有价值的见解，细胞如何调节其动态信号行为，根据其在组织内的定位。在目标1中，我们将创建具有预定义几何形状的3D生物工程组织，以确定几何约束如何影响乳腺上皮组织侵入的能力。在目标2中，我们将使用荧光报告分子进行延时共聚焦显微镜分析，以跟踪这些生物工程组织中所有细胞的ERK信号动力学，并确定细胞形状和定位如何在入侵过程中调节ERK信号。我们还将测试ERK通路的激活是否足以通过使用允许空间和时间控制的光遗传学工具来诱导或抑制侵袭。在目标3中，我们将应用最近开发的光遗传学工具来诱导3D组织内特定细胞的细胞收缩，以确定机械力对ERK信号传导和侵袭的影响。总之，这项工作将揭示组织几何形状，机械力和ERK信号动力学如何相互作用以驱动肿瘤侵袭，从而定义早期乳腺癌转移级联的关键步骤，这些步骤可以在治疗上中断。此外，这项工作将提供新的途径来监测和操纵肿瘤细胞内的信号传导，这广泛适用于其他类型的癌症。