Immunology and Reproduction Biology Laboratory & State Key Laboratory of Analytical Chemistry for Life Science, Medical School, Nanjing University, Nanjing, 210093, China.
Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing, 210093, China.
Theranostics. 2021 Jan 9;11(7):3244-3261. doi: 10.7150/thno.54217. eCollection 2021.
(Myo)fibroblasts are the ultimate effector cells responsible for the production of collagen within alveolar structures, a core phenomenon in the pathogenesis of idiopathic pulmonary fibrosis (IPF). Although (myo)fibroblast-targeted therapy holds great promise for suppressing the progression of IPF, its development is hindered by the limited drug delivery efficacy to (myo)fibroblasts and the vicious circle of (myo)fibroblast activation and evasion of apoptosis. Here, a dual small interfering RNA (siRNA)-loaded delivery system of polymeric micelles is developed to suppress the development of pulmonary fibrosis via a two-arm mechanism. The micelles are endowed with (myo)fibroblast-targeting ability by modifying the Fab' fragment of the anti-platelet-derived growth factor receptor-α (PDGFRα) antibody onto their surface. Two different sequences of siRNA targeting protein tyrosine phosphatase-N13 (PTPN13, a promoter of the resistance of (myo)fibroblasts to Fas-induced apoptosis) and NADPH oxidase-4 (NOX4, a key regulator for (myo)fibroblast differentiation and activation) are loaded into micelles to inhibit the formation of fibroblastic foci. We demonstrate that Fab'-conjugated dual siRNA-micelles exhibit higher affinity to (myo)fibroblasts in fibrotic lung tissue. This Fab'-conjugated dual siRNA-micelle can achieve remarkable antifibrotic effects on the formation of fibroblastic foci by, on the one hand, suppressing (myo)fibroblast activation via siRNA-induced knockdown of NOX4 and, on the other hand, sensitizing (myo)fibroblasts to Fas-induced apoptosis by siRNA-mediated PTPN13 silencing. In addition, this (myo)fibroblast-targeting siRNA-loaded micelle did not induce significant damage to major organs, and no histopathological abnormities were observed in murine models. The (myo)fibroblast-targeting dual siRNA-loaded micelles offer a potential strategy with promising prospects in molecular-targeted fibrosis therapy.
肌成纤维细胞是肺泡结构中产生胶原的最终效应细胞，这是特发性肺纤维化（IPF）发病机制的核心现象。虽然肌成纤维细胞靶向治疗具有抑制 IPF 进展的巨大潜力，但由于向肌成纤维细胞输送药物的效率有限，以及肌成纤维细胞的激活和逃避细胞凋亡的恶性循环，其发展受到了阻碍。 在这里，开发了一种负载双重小干扰 RNA（siRNA）的聚合物胶束递药系统，通过双管齐下的机制来抑制肺纤维化的发展。通过将抗血小板衍生生长因子受体-α（PDGFRα）抗体的 Fab'片段修饰在胶束表面，使胶束具有肌成纤维细胞靶向能力。将两种不同序列的靶向蛋白酪氨酸磷酸酶-N13（PTPN13，肌成纤维细胞对 Fas 诱导的细胞凋亡的抗性的启动子）和 NADPH 氧化酶-4（NOX4，肌成纤维细胞分化和激活的关键调节剂）的 siRNA 负载到胶束中，以抑制成纤维细胞灶的形成。 我们证明，在纤维化的肺组织中，与 Fab'缀合的双重 siRNA-胶束对肌成纤维细胞具有更高的亲和力。这种 Fab'缀合的双重 siRNA-胶束通过一方面通过 siRNA 诱导的 NOX4 敲低抑制肌成纤维细胞的激活，另一方面通过 siRNA 介导的 PTPN13 沉默使肌成纤维细胞对 Fas 诱导的细胞凋亡敏感，从而对成纤维细胞灶的形成产生显著的抗纤维化作用。此外，这种肌成纤维细胞靶向的 siRNA 负载胶束不会对主要器官造成明显的损伤，在小鼠模型中也没有观察到组织病理学异常。 肌成纤维细胞靶向的双重 siRNA 负载胶束为靶向纤维化治疗提供了一种有前途的策略。
