Extracellular Hsp90 Binds to and Aligns Collagen-1 to Enhance Breast Cancer Cell Invasiveness.
Extracellular Hsp90 Binds to and Aligns Collagen-1 to Enhance Breast Cancer Cell Invasiveness.
复制标题
细胞外HSP90与胶原-1结合并对齐以增强乳腺癌细胞的侵入性。
DOI：
10.3390/cancers15215237
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发表时间：
2023-10-31
期刊：
影响因子：
5.2
通讯作者：
Jay, Daniel G.
中科院分区：
文献类型：
--
作者：
Singh, Pragya;Ramanathan, Varshini;Zhang, Yang;Georgakoudi, Irene;Jay, Daniel G.
关键词：
乳腺癌细胞分泌Hsp 90，这是一种在细胞内调节数百种蛋白质功能的蛋白质，但在细胞外，细胞外Hsp 90（eHsp 90）可以激活促进侵袭的蛋白质子集，这是转移的第一步。在小鼠模型中阻断eHsp 90可以抑制转移，我们试图了解这是如何发生的。先前的研究主要集中在eHsp 90在原发性肿瘤附近的癌症侵袭中，特别是其在上皮隔室外侵袭中的作用。然而，eHsp 90在细胞穿过上皮区室的边界后跨越扩展的结缔组织的癌症侵袭中的作用仍然未知。我们在这里表明，eHsp 90直接结合并排列胶原蛋白-1纤维，胶原蛋白-1纤维是结缔组织的主要结构成分，当排列时，形成允许有效癌症迁移的高速公路。我们的研究表明，热休克蛋白90二聚体，在其开放状态下，结合胶原蛋白-1分子排列的纤维，这导致增强乳腺癌通过胶原蛋白-1基质的侵袭。了解这一点可以帮助我们提出实验来测试eHsp 90抑制剂用于治疗转移性乳腺癌。癌细胞分泌的eHsp 90结合并激活肿瘤微环境中对癌症侵袭至关重要的蛋白质。因此，靶向eHsp 90可以抑制侵袭，防止转移-癌症相关死亡率的主要原因。先前的eHsp 90研究仅关注其通过2D基底膜（BM）在癌症侵袭中的作用，2D基底膜是一种细胞外基质（ECM）形式，其排列在上皮隔室中。然而，其通过3D间质基质（IM）（BM之外的ECM）在癌症侵袭中的作用仍未被探索。使用胶原蛋白-1结合试验和二次谐波产生（SHG）成像，我们证明了eHsp 90直接结合和排列胶原蛋白-1纤维，IM的主要成分。此外，我们表明，eHsp 90增强胶原蛋白-1的乳腺癌细胞的侵袭在Transwell测定。使用Hsp 90构象突变体和抑制剂，我们建立了Hsp 90二聚体通过其N-结构域与胶原蛋白1结合。我们还证明，虽然胶原蛋白-1的结合和排列不受归因于N-结构域的Hsp 90的ATP酶活性的影响，但其开放构象对于增加胶原蛋白-1的排列和促进乳腺癌细胞侵袭至关重要。这些发现揭示了eHsp 90通过IM在侵袭中的新作用，并为抑制Hsp 90以抑制侵袭和转移的潜在治疗方法提供了有价值的机制见解。
Breast cancer cells secrete Hsp90, a protein that, inside of cells, regulates the function of hundreds of proteins, but outside of cells, extracellular Hsp90 (eHsp90) can activate a subset of proteins that promote invasion, the first step of metastasis. Blocking eHsp90 in mouse models inhibits metastasis, and we sought to understand how this occurs. Prior studies have predominantly focused on eHsp90 in cancer invasion within the immediate vicinity of the primary tumor, specifically its role in invading outside the epithelial compartment. However, eHsp90’s role in cancer invasion across the extended connective tissue after the cells have crossed the boundary of the epithelial compartment remains unknown. We show here that eHsp90 directly binds to and aligns Collagen-1 fibers, a major structural component of connective tissues, which, when aligned, form highways that allow efficient cancer migration. Our study suggests that the Hsp90 dimer, in its open state, binds to Collagen-1 molecules to align the fibers, which results in enhanced breast cancer invasion through the Collagen-1 matrix. Knowing this could help us propose experiments to test eHsp90 inhibitors for therapeutically targeting metastatic breast cancer. Cancer cell-secreted eHsp90 binds and activates proteins in the tumor microenvironment crucial in cancer invasion. Therefore, targeting eHsp90 could inhibit invasion, preventing metastasis—the leading cause of cancer-related mortality. Previous eHsp90 studies have solely focused on its role in cancer invasion through the 2D basement membrane (BM), a form of extracellular matrix (ECM) that lines the epithelial compartment. However, its role in cancer invasion through the 3D Interstitial Matrix (IM), an ECM beyond the BM, remains unexplored. Using a Collagen-1 binding assay and second harmonic generation (SHG) imaging, we demonstrate that eHsp90 directly binds and aligns Collagen-1 fibers, the primary component of IM. Furthermore, we show that eHsp90 enhances Collagen-1 invasion of breast cancer cells in the Transwell assay. Using Hsp90 conformation mutants and inhibitors, we established that the Hsp90 dimer binds to Collagen-1 via its N-domain. We also demonstrated that while Collagen-1 binding and alignment are not influenced by Hsp90’s ATPase activity attributed to the N-domain, its open conformation is crucial for increasing Collagen-1 alignment and promoting breast cancer cell invasion. These findings unveil a novel role for eHsp90 in invasion through the IM and offer valuable mechanistic insights into potential therapeutic approaches for inhibiting Hsp90 to suppress invasion and metastasis.
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