Dynamic modulation of matrix adhesiveness induces epithelial-to-mesenchymal transition in prostate cancer cells in 3D.
Dynamic modulation of matrix adhesiveness induces epithelial-to-mesenchymal transition in prostate cancer cells in 3D.
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DOI：
10.1016/j.biomaterials.2023.122180
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发表时间：
2023-05
期刊：
影响因子：
14
通讯作者：
Mugdha Pol;Hanyuan Gao;He Zhang;Olivia J George;Joseph M Fox;Xinqiao Jia
中科院分区：
文献类型：
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作者：
Mugdha Pol;Hanyuan Gao;He Zhang;Olivia J George;Joseph M Fox;Xinqiao Jia
动态呈现细胞导向线索的合成基质是开发生理相关的体外肿瘤模型所需要的。为了模拟前列腺癌进展和转移的目标，我们设计了一种可调的基于透明质酸的水凝胶平台，其具有蛋白酶可降解和细胞粘附特性，采用生物正交四嗪与应变烯烃连接。合成基质首先通过缓慢的四嗪-异辛烯反应制造，然后通过扩散控制方法使用反式环辛烯进行临时修改，反式环辛烯是一种激烈的亲二烯体，与四嗪反应速度异常快。包封的DU 145前列腺癌单细胞在培养7天后自发形成多细胞类肿瘤，通过共价标记细胞粘附性RGD肽的合成基质的原位修饰诱导类肿瘤解压缩和细胞突起的发展。RGD标记不损害整体细胞活力，也不诱导细胞凋亡。为了响应增加的基质粘附，DU 145细胞动态地放松细胞-细胞粘附并加强细胞-基质相互作用以促进侵袭性表型。通过免疫细胞化学和基因表达分析表征的3D培养物表明，细胞通过间充质样迁移侵入基质，主要间充质标志物上调，上皮标志物下调。类瘤形成coronin阳性的侵入伪足样结构，表明活跃的基质重塑。总的来说，工程肿瘤模型可用于识别潜在的分子靶点和测试药理学抑制剂，从而加速癌症治疗创新策略的设计。
Synthetic matrices with dynamic presentation of cell guidance cues are needed for the development of physiologically relevant in vitro tumor models. Towards the goal of mimicking prostate cancer progression and metastasis, we engineered a tunable hyaluronic acid-based hydrogel platform with protease degradable and cell adhesive properties employing bioorthogonal tetrazine ligation with strained alkenes. The synthetic matrix was first fabricated via a slow tetrazine-norbornene reaction, then temporally modified via a diffusion-controlled method usingtrans-cyclooctene, a fierce dienophile that reacts with tetrazine with an unusually fast rate. The encapsulated DU145 prostate cancer single cells spontaneously formed multicellular tumoroids after 7 days of culture.In situmodification of the synthetic matrix via covalent tagging of cell adhesive RGD peptide induced tumoroid decompaction and the development of cellular protrusions. RGD tagging did not compromise the overall cell viability, nor did it induce cell apoptosis. In response to increased matrix adhesiveness, DU145 cells dynamically loosen cell-cell adhesion and strengthen cell-matrix interactions to promote an invasive phenotype. Characterization of the 3D cultures by immunocytochemistry and gene expression analyses demonstrated that cells invaded into the matrix via a mesenchymal like migration, with upregulation of major mesenchymal markers, and down regulation of epithelial markers. The tumoroids formed cortactin positive invadopodia like structures, indicating active matrix remodeling. Overall, the engineered tumor model can be utilized to identify potential molecular targets and test pharmacological inhibitors, thereby accelerating the design of innovative strategies for cancer therapeutics.