De Luca Giada, Petrillo Gianluca, Scognamiglio Iolanda, Pane Katia, Cocca Lorenza, Roscigno Giuseppina, Mascolo Martina, Pignataro Claudia, Verde Sara, Fraticelli Aurelia, Fiore Danilo, Affinito Alessandra, Nuzzo Silvia, Minic Zoran, De Micco Francesca, Thomas Guglielmo, Franzese Monica, Berezovski Maxim V, Fedele Monica, Condorelli Gerolama, Quintavalle Cristina
Institute of Endotypes in Oncology, Metabolism and Immunology "G. Salvatore" (IEOMI), Consiglio Nazionale delle Ricerche (CNR), Naples, Italy.
Department of Molecular Medicine and Medical Biotechnology, "Federico II" University of Naples, V. Tommaso de Amicis 95, Naples, 80131, Italy.
Cell Mol Life Sci. 2025 Jul 25;82(1):287. doi: 10.1007/s00018-025-05781-y.
The intricate interplay between epithelial and fibroblast cells within the tumor microenvironment plays a crucial role in driving triple-negative breast cancer progression. This crosstalk involves the exchange of various signaling molecules, including growth factors, cytokines, extracellular matrix components, and extracellular vesicles. Recently, we demonstrated that triple-negative breast cancer extracellular vesicles carry and release a specific combination of miRs, including miR-185-5p, miR-652-5p, and miR-1246 (from here on, referred as combo-miRs), into normal fibroblasts, effectively reprogramming them into cancer-associated fibroblasts. Here, we show that the conditioned medium from the fibroblasts activated by combo-miRs exerts a pro-tumorigenic effect on epithelial cells, enhancing the viability and migratory potential while driving increased invasiveness in patient-derived breast cancer organoids. A proteomic analysis of conditioned medium from combo-miRs activated fibroblasts revealed 76 significantly upregulated secreted proteins compared to control. Bioinformatic analysis identified the transcriptional factor PATZ1 as a potential regulator of the 12 most highly upregulated proteins. Consistently, in-silico predictions and in vitro experiments confirmed that PATZ1 is a direct target of miR-185-5p and miR-652-5p. The downregulation of PATZ1 by these miRNAs led to increased levels of the secreted proteins in the conditioned medium from combo-miRs activated fibroblasts. Furthermore, the conditioned medium from PATZ1-knockout mesenchymal embryonic fibroblasts and normal fibroblasts with silenced PATZ1 similarly enhanced the migratory potential of MCF10A cells, further supporting the critical role of PATZ1 in regulating tumor-promoting mechanisms. These findings provide valuable insights into the dynamics of the TME in TNBC, highlighting combo-miRs and PATZ1 as promising targets for future therapeutic interventions.
肿瘤微环境中上皮细胞和成纤维细胞之间复杂的相互作用在驱动三阴性乳腺癌进展中起着关键作用。这种相互作用涉及多种信号分子的交换，包括生长因子、细胞因子、细胞外基质成分和细胞外囊泡。最近，我们证明三阴性乳腺癌细胞外囊泡携带并向正常成纤维细胞释放特定组合的微小RNA（miR），包括miR-185-5p、miR-652-5p和miR-1246（以下简称组合微小RNA），有效地将它们重编程为癌症相关成纤维细胞。在此，我们表明，由组合微小RNA激活的成纤维细胞的条件培养基对上皮细胞具有促肿瘤作用，增强了其活力和迁移潜力，同时在患者来源的乳腺癌类器官中增加了侵袭性。对组合微小RNA激活的成纤维细胞的条件培养基进行蛋白质组分析发现，与对照组相比，有76种分泌蛋白显著上调。生物信息学分析确定转录因子PATZ1是12种上调程度最高的蛋白质的潜在调节因子。一致地，计算机模拟预测和体外实验证实PATZ1是miR-185-5p和miR-652-5p的直接靶点。这些微小RNA对PATZ1的下调导致组合微小RNA激活的成纤维细胞的条件培养基中分泌蛋白水平升高。此外，来自PATZ1基因敲除的间充质胚胎成纤维细胞和PATZ1沉默的正常成纤维细胞的条件培养基同样增强了MCF10A细胞的迁移潜力，进一步支持了PATZ1在调节肿瘤促进机制中的关键作用。这些发现为三阴性乳腺癌中肿瘤微环境的动态变化提供了有价值的见解，突出了组合微小RNA和PATZ1作为未来治疗干预的有希望的靶点。
