Varying the RGD concentration on a hyaluronic acid hydrogel influences dormancy versus proliferation in brain metastatic breast cancer cells
Varying the RGD concentration on a hyaluronic acid hydrogel influences dormancy versus proliferation in brain metastatic breast cancer cells
复制标题
改变透明质酸水凝胶上的 RGD 浓度会影响脑转移性乳腺癌细胞的休眠与增殖
DOI：
10.1002/jbm.a.37651
复制
发表时间：
2023
影响因子：
4.9
通讯作者：
Rao, Shreyas S.
中科院分区：
文献类型：
--
作者：
Goodarzi, Kasra;Lane, Rachel;Rao, Shreyas S.
大多数乳腺癌死亡是由于癌细胞转移到远处器官造成的。特别是，脑转移的侵袭性很强，存活率极低。转移到大脑的乳腺癌细胞可以进入休眠状态，这使它们能够逃脱死亡。脑微环境提供生物物理、生化和细胞线索，并在决定处于休眠状态的癌细胞的命运中发挥重要作用。然而，这些线索如何影响休眠仍然知之甚少。在这里，我们使用硬度为~0.4RGD的透明质酸(HA)水凝胶作为体外仿生平台，研究生化信号，特别是RGD浓度的变化，对 -MB-231BR脑转移癌细胞休眠和增殖的影响。我们将不同浓度的RGD肽(0、1、2或4 mg/mL)应用于HA水凝胶表面，并使用荧光标记的RGD肽证实了不同程度的表面功能化。功能化后，将10,000个MDAMB-231BR细胞接种于水凝胶上，培养5 d。我们发现，随着RGD浓度的增加，细胞形态发生了变化，细胞从圆形转变为纺锤形，细胞铺展面积增加。此外，RGD浓度的增加会导致细胞增殖的增加。用磷酸化细胞外信号调节激酶1/2(p-ERK)与磷酸化p38(p-p38)的比值来评估细胞休眠，与RGD浓度较高的水凝胶相比，不含RGD的水凝胶和RGD浓度最低的水凝胶中的p-ERK/p38阳性率显著降低。我们还证明了HA水凝胶诱导的细胞休眠是可逆的。最后，我们在我们的水凝胶平台中证明了β-1整合素参与了细胞表型的调节。总体而言，我们的结果提供了对生化信号在调节脑转移乳腺癌细胞休眠和增殖中的作用的洞察。
A majority of breast cancer deaths occur due to metastasis of cancer cells to distant organs. In particular, brain metastasis is very aggressive with an extremely low survival rate. Breast cancer cells that metastasize to the brain can enter a state of dormancy, which allows them to evade death. The brain microenvironment provides biophysical, biochemical, and cellular cues, and plays an important role in determining the fate of dormant cancer cells. However, how these cues influence dormancy remains poorly understood. Herein, we employed hyaluronic acid (HA) hydrogels with a stiffness of ~0.4 kPa as an in vitro biomimetic platform to investigate the impact of biochemical cues, specifically alterations in RGD concentration, on dormancy versus proliferation in MDA‐MB‐231Br brain metastatic breast cancer cells. We applied varying concentrations of RGD peptide (0, 1, 2, or 4 mg/mL) to HA hydrogel surfaces and confirmed varying degrees of surface functionalization using a fluorescently labeled RGD peptide. Post functionalization, ~10,000 MDA‐MB‐231Br cells were seeded on top of the hydrogels and cultured for 5 days. We found that an increase in RGD concentration led to changes in cell morphology, with cells transitioning from a rounded to spindle‐like morphology as well as an increase in cell spreading area. Also, an increase in RGD concentration resulted in an increase in cell proliferation. Cellular dormancy was assessed using the ratio of phosphorylated extracellular signal‐regulated kinase 1/2 (p‐ERK) to phosphorylated p38 (p‐p38) positivity, which was significantly lower in hydrogels without RGD and in hydrogels with lowest RGD concentration compared to hydrogels functionalized with higher RGD concentration. We also demonstrated that the HA hydrogel‐induced cellular dormancy was reversible. Finally, we demonstrated the involvement of β1 integrin in mediating cell phenotype in our hydrogel platform. Overall, our results provide insight into the role of biochemical cues in regulating dormancy versus proliferation in brain metastatic breast cancer cells.