Cancer under pressure: Mechanisms of adaptation to compressive stress

项目总结 物理压力对癌症生物学至关重要，但它的影响仍然知之甚少。什么时候 实体瘤生长在周围组织内，它们会产生压应力。考虑到细胞的进化 要在稳定的机械环境中发挥作用，即使是压力的轻微变化也会扰乱生理。正常细胞 当压力增加时，早期癌细胞就会停止生长。相比之下，在晚期癌症中，压迫 可以改变细胞行为，推动癌细胞向其他器官迁移或对化疗产生抵抗力。 心理治疗。这种不同意味着癌细胞以某种方式适应了物理压力。工具的缺乏已经减缓了 了解压缩、细胞物理性质和癌症之间关系的进展 行为。我们开发了两种新技术来克服这一限制：首先，我们创造了一种基因，可以使 细胞产生稳定的荧光纳米颗粒，作为细胞内物理变化的警示 属性。其次，我们开发了微流控装置来控制压应力，无论是快是慢， 同时保持恒定的化学环境。我们将结合这些创新来测试最重要的 一种假说认为，赋予机械压缩抵抗力的突变使胰腺癌细胞能够 适应他们的高压环境，并推动他们的致癌进化。目标1：我们将确定如何 压迫对野生型和突变型胰腺细胞的影响不同。我们将使用宝石纳米颗粒 为了量化身体对压力的反应，并检验致癌基因突变改变 对压力的生理和生理反应。目标2：我们将确定压缩对 相分离。我们将研究在压力作用下细胞体积减少会导致细胞内 应力颗粒的相分离变得更加明显。我们将评估分子拥挤作为这些问题的一种机制 效果。我们将确定应力颗粒形成对机械适应和药物再适应的重要性。 抵抗。目的3：我们将确定压力适应的遗传机制。我们会跟进之前的 赋予抗压性的亚胺类突变体，使用CRISPR修饰物筛选来确定机制- 适应的习性。我们将过度表达已知的癌基因，以寻找进一步的适应途径。 我们的基因纳米粒子和微流控方法的创新组合，以及我们的专业知识 桥梁生物物理学、机械生物学和细胞生物学使我们唯一有资格连接压缩， 细胞的物理化学性质和癌症生理学。我们的研究承诺揭示关键的网络文化- 对于癌细胞在压力下的生长和存活至关重要。了解这些适应机制 有希望提出利用高成分的肿瘤的异常机械性能的治疗方法。 压力过大。

Project summary Physical pressure is fundamentally important for cancer biology, but its effects remain poorly understood. When solid tumors grow confined within surrounding tissue, they build up compressive stress. Given that cells evolved to function in a stable mechanical environment, even slight changes in pressure perturb physiology. Normal cells and early stage cancer cells stop growing when pressure builds up. In contrast, in advanced cancer, compression can change cellular behavior to drive migration of cancer cells to other organs or confer resistance to chemo- therapy. This difference implies that cancer cells somehow adapt to physical pressure. A lack of tools has slowed progress in understanding the relationships between compression, the physical properties of cells, and cancer behavior. We developed two new technologies to overcome this limitation: First, we created a gene that enables cells to produce a steady supply of fluorescent nanoparticles that act as tell-tales for shifts in intracellular physical properties. Second, we developed microfluidic devices to control compressive stress, either quickly or slowly, while maintaining a constant chemical environment. We will combine these innovations to test the overarching hypothesis that mutations that confer resistance to mechanical compression enable pancreatic cancer cells to adapt to their high-pressure environment and drive their oncogenic evolution. Aim 1: We will determine how compression differentially impacts wildtype and mutant pancreatic cells. We will use GEM nanoparticles to quantify the physical response to pressure and test the hypothesis that oncogenic mutations alter both the physical and physiological response to pressure. Aim 2: We will determine the effects of compression on phase separation. We will investigate the hypothesis that decreased cell volume under pressure leads to in- creased phase separation of stress granules. We will evaluate molecular crowding as a mechanism for these effects. We will determine the importance of stress granule formation for mechanical adaptation and drug re- sistance. Aim 3: We will determine genetic mechanisms of pressure adaptation. We will follow up on pre- liminary mutants that confer resistance to compression, using a CRISPR modifier screen to determine mecha- nisms of adaptation. We will overexpress known oncogenes to find further adaptation pathways. Our innovative combination of genetic nanoparticles and microfluidic approaches, and our expertise that bridges biophysics, mechanobiology and cell biology make us uniquely qualified to connect compression, the physicochemical properties of cells, and cancer physiology. Our studies promise to reveal key network pertur- bations essential to cancer cell growth and survival under pressure. Understanding these adaptive mechanisms promises to suggest treatments that exploit the aberrant mechanical properties of tumors caused by high com- pressive stress.