Therapeutic Effect of Cyclin-Dependent Kinase 4/6 Inhibitor on Dermal Fibrosis in Murine Models of Systemic Sclerosis.
Therapeutic Effect of Cyclin-Dependent Kinase 4/6 Inhibitor on Dermal Fibrosis in Murine Models of Systemic Sclerosis.
复制标题
细胞周期蛋白依赖性激酶 4/6 抑制剂对系统性硬化症小鼠模型真皮纤维化的治疗作用。
DOI：
10.1002/art.42042
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Kohsaka H.
中科院分区：
文献类型：
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作者：
Yamamoto A;Saito T;Hosoya T;Kawahata K;Asano Y;Sato S;Mizoguchi F;Yasuda S;Kohsaka H.
系统性硬化症（systemic sclerosis，SSc）的组织学特征之一是真皮肌成纤维细胞数量增加，转化生长因子β（transforming growth factor β，TGFβ）在促进成纤维细胞向肌成纤维细胞分化，导致真皮纤维化中起关键作用。本研究旨在：1）检查用细胞周期蛋白依赖性激酶4/6（CDK 4/6）抑制剂抑制细胞周期是否会抑制成纤维细胞的增殖及其向肌成纤维细胞的分化，以及2）评估CDK 4/6抑制剂作为单一疗法或与TGFβ受体（TGFβR）抑制剂联合给药的治疗效果，方法将从SSc患者皮肤中获得的成纤维细胞在存在或不存在TGFβ的情况下培养。使用溴脱氧尿苷摄取试验以及免疫荧光和免疫印迹分析，检查了palbociclib（一种CDK 4/6抑制剂）对成纤维细胞增殖和TGFβ诱导分化为肌成纤维细胞的影响。HOCl和博来霉素诱导的皮肤纤维化的小鼠模型用于研究CDK 4/6抑制剂对皮肤纤维化的影响，CDK 4/6抑制剂治疗作为单药治疗或与galunisertib（一种TGFβR抑制剂）联合给药。结果向细胞培养物中加入CDK 4/6抑制剂可抑制人皮肤SSc成纤维细胞的增殖及其TGFβ诱导的肌成纤维细胞分化，而不抑制典型和非典型TGFβ信号。在皮肤纤维化的鼠模型中，用CDK 4/6抑制剂治疗小鼠降低了真皮厚度和胶原蛋白含量，以及真皮成纤维细胞增殖和肌成纤维细胞的数量。CDK 4/6抑制剂和TGFβR抑制剂的联合治疗产生了相加的抗纤维化作用。从机制上讲，CDK 4/6抑制剂抑制了细胞通讯网络2和钙粘蛋白-11的表达，这是在纤维化的发展和进展中起重要作用的蛋白质。结论这项研究的结果表明，CDK 4/6抑制剂对真皮纤维化的治疗效果时，作为单一药物或与TGFβR抑制剂联合给药。CDK 4/6抑制剂，包括本研究中使用的palbociclib，可能代表治疗SSc的新型药物，如果与TGFβR抑制剂联合使用，可能会增加疗效。
ObjectiveOne of the histologic characteristics of systemic sclerosis (SSc) is an increased number of dermal myofibroblasts, and transforming growth factor β (TGFβ) plays a crucial role in the promotion of myofibroblast differentiation from fibroblasts, leading to dermal fibrosis. This study was undertaken to 1) examine whether inhibition of the cell cycle with a cyclin‐dependent kinase 4/6 (CDK4/6) inhibitor suppresses the proliferation of fibroblasts and their differentiation into myofibroblasts, and 2) assess the therapeutic effects of a CDK4/6 inhibitor, administered as monotherapy or in combination with a TGFβ receptor (TGFβR) inhibitor, on dermal fibrosis in murine models of SSc.MethodsFibroblasts obtained from the skin of patients with SSc were cultured in the presence or absence of TGFβ. The effects of palbociclib, a CDK4/6 inhibitor, on fibroblast proliferation and TGFβ‐induced differentiation into myofibroblasts were examined using bromodeoxyuridine uptake assays as well as immunofluorescence and immunoblotting analyses. Murine models of HOCl‐ and bleomycin‐induced dermal fibrosis were used to study the effect of a CDK4/6 inhibitor on dermal fibrosis, with the CDK4/6 inhibitor treatment administered as monotherapy or in combination with galunisertib, a TGFβR inhibitor.ResultsAddition of a CDK4/6 inhibitor to the cell cultures suppressed the proliferation of human dermal SSc fibroblasts and their TGFβ‐induced differentiation into myofibroblasts, without inhibiting canonical and noncanonical TGFβ signals. In murine models of dermal fibrosis, treatment of mice with a CDK4/6 inhibitor decreased dermal thickness and collagen content, as well as dermal fibroblast proliferation and the numbers of myofibroblasts. Combination therapy with the CDK4/6 inhibitor and TGFβR inhibitor resulted in additive antifibrotic effects. Mechanistically, the CDK4/6 inhibitor suppressed the expression of cellular communication network 2 and cadherin‐11, which are proteins that have important roles in the development and progression of fibrosis.ConclusionResults of this study demonstrate the therapeutic effect of a CDK4/6 inhibitor on dermal fibrosis when administered as monotherapy or in combination with a TGFβR inhibitor. CDK4/6 inhibitors, including palbociclib used in the present study, may represent novel agents for the treatment of SSc, which, if used in combination with a TGFβR inhibitor, might result in increased efficacy.