Tanaka Hiroyoshi Y, Nakazawa Takuya, Miyazaki Takuya, Cabral Horacio, Masamune Atsushi, Kano Mitsunobu R
Department of Pharmaceutical Biomedicine, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University, 1-1-1 Tsushima-naka, Kita-ku, Okayama-shi, Okayama 700-8530, Japan.
Department of Pharmaceutical Biomedicine, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University, 1-1-1 Tsushima-naka, Kita-ku, Okayama-shi, Okayama 700-8530, Japan.
J Control Release. 2024 May;369:283-295. doi: 10.1016/j.jconrel.2024.03.041. Epub 2024 Mar 29.
Pancreatic cancer is characterized by a densely fibrotic stroma. The fibrotic stroma hinders the intratumoral penetration of nanomedicine and diminishes therapeutic efficacy. Fibrosis is characterized by an abnormal organization of extracellular matrix (ECM) components, namely the abnormal deposition and/or orientation of collagen and fibronectin. Abnormal ECM organization is chiefly driven by pathological signaling in pancreatic stellate cells (PSCs), the main cell type involved in fibrogenesis. However, whether targeting signaling pathways involved in abnormal ECM organization improves the intratumoral penetration of nanomedicines is unknown. Here, we show that targeting transforming growth factor-β (TGFβ)/Rho-associated kinase (ROCK) 1/2 signaling in PSCs normalizes ECM organization and concomitantly improves macromolecular permeability of the fibrotic stroma. Using a 3-dimensional cell culture model of the fibrotic pancreatic cancer microenvironment, we found that pharmacological inhibition of TGFβ or ROCK1/2 improves the permeation of various macromolecules. By using an isoform-specific pharmacological inhibitor and siRNAs, we show that targeting ROCK2, but not ROCK1, alone is sufficient to normalize ECM organization and improve macromolecular permeability. Moreover, we found that ROCK2 inhibition/knockdown attenuates Yes-associated protein (YAP) nuclear localization in fibroblasts co-cultured with pancreatic cancer cells in 3D. Finally, pharmacological inhibition or siRNA-mediated knockdown of YAP normalized ECM organization and improved macromolecular permeability. Our results together suggest that the TGFβ/ROCK2/YAP signaling axis may be therapeutically targeted to normalize ECM organization and improve macromolecular permeability to augment therapeutic efficacy of nanomedicines in pancreatic cancer.
胰腺癌的特征是存在致密的纤维化基质。这种纤维化基质会阻碍纳米药物在肿瘤内的渗透，并降低治疗效果。纤维化的特征是细胞外基质（ECM）成分的异常组织，即胶原蛋白和纤连蛋白的异常沉积和/或排列。ECM的异常组织主要由胰腺星状细胞（PSC）中的病理信号驱动，PSC是参与纤维化形成的主要细胞类型。然而，靶向参与ECM异常组织的信号通路是否能改善纳米药物在肿瘤内的渗透尚不清楚。在这里，我们表明，靶向PSC中的转化生长因子-β（TGFβ）/Rho相关激酶（ROCK）1/2信号通路可使ECM组织正常化，并同时提高纤维化基质的大分子通透性。使用纤维化胰腺癌微环境的三维细胞培养模型，我们发现对TGFβ或ROCK1/2进行药理抑制可改善各种大分子的渗透。通过使用亚型特异性药理抑制剂和小干扰RNA（siRNA），我们表明单独靶向ROCK2而非ROCK1足以使ECM组织正常化并提高大分子通透性。此外，我们发现抑制/敲低ROCK2会减弱与胰腺癌三维共培养的成纤维细胞中Yes相关蛋白（YAP）的核定位。最后，对YAP进行药理抑制或siRNA介导的敲低可使ECM组织正常化并提高大分子通透性。我们的研究结果共同表明，TGFβ/ROCK2/YAP信号轴可能是一个治疗靶点，可使ECM组织正常化并提高大分子通透性，从而增强纳米药物在胰腺癌中的治疗效果。
