Research Laboratory, Dental School, University of Sevilla, Seville, Spain; Department of Periodontology, Dental School, University of Sevilla, Seville, Spain.
Research Laboratory, Dental School, University of Sevilla, Seville, Spain.
J Allergy Clin Immunol. 2018 Oct;142(4):1131-1143.e7. doi: 10.1016/j.jaci.2018.01.018. Epub 2018 Feb 2.
Cathepsin C (CatC) is a lysosomal enzyme involved in activation of serine proteases from immune and inflammatory cells. Several loss-of-function mutations in the CatC gene have been shown to be the genetic mark of Papillon-Lefèvre syndrome (PLS), a rare autosomal recessive disease characterized by severe early-onset periodontitis, palmoplantar hyperkeratosis, and increased susceptibility to infections. Deficiencies or dysfunction in other cathepsin family proteins, such as cathepsin B or D, have been associated with autophagic and lysosomal disorders.
Here we characterized the basis for autophagic dysfunction in patients with PLS by analyzing skin fibroblasts derived from patients with several mutations in the CatC gene and reduced enzymatic activity.
Skin fibroblasts were isolated from patients with PLS assessed by using genetic analysis. Authophagic flux dysfunction was evaluated by examining accumulation of p62/SQSTM1 and a bafilomycin assay. Ultrastructural analysis further confirmed abnormal accumulation of autophagic vesicles in mutant cells. A recombinant CatC protein was produced by a baculovirus system in insect cell cultures.
Mutant fibroblasts from patients with PLS showed alterations in oxidative/antioxidative status, reduced oxygen consumption, and a marked autophagic dysfunction associated with autophagosome accumulation. These alterations were accompanied by lysosomal permeabilization, cathepsin B release, and NLR family pyrin domain containing 3 (NLRP3) inflammasome activation. Treatment of mutant fibroblasts with recombinant CatC improved cell growth and autophagic flux and partially restored lysosomal permeabilization.
Our data provide a novel molecular mechanism underlying PLS. Impaired autophagy caused by insufficient lysosomal function might represent a new therapeutic target for PLS.
组织蛋白酶 C(CatC)是一种参与免疫和炎症细胞中丝氨酸蛋白酶激活的溶酶体酶。已经发现 CatC 基因的几种功能丧失突变是 Papillon-Lefèvre 综合征(PLS)的遗传标志,这是一种罕见的常染色体隐性遗传病,其特征是严重的早发性牙周炎、手掌足底过度角化和对感染的易感性增加。其他组织蛋白酶家族蛋白(如组织蛋白酶 B 或 D)的缺乏或功能障碍与自噬和溶酶体疾病有关。
通过分析来自 CatC 基因突变和酶活性降低的患者的皮肤成纤维细胞,我们研究了 PLS 患者自噬功能障碍的基础。
通过遗传分析评估来自 PLS 患者的皮肤成纤维细胞。通过检查 p62/SQSTM1 的积累和巴弗洛霉素测定来评估自噬通量功能障碍。超微结构分析进一步证实了突变细胞中异常的自噬小体积累。通过杆状病毒系统在昆虫细胞培养物中产生重组 CatC 蛋白。
来自 PLS 患者的突变成纤维细胞显示氧化/抗氧化状态改变、耗氧量降低以及与自噬体积累相关的明显自噬功能障碍。这些改变伴随着溶酶体通透性增加、组织蛋白酶 B 释放和 NLR 家族 pyrin 结构域包含 3(NLRP3)炎症小体激活。用重组 CatC 处理突变细胞可改善细胞生长和自噬通量,并部分恢复溶酶体通透性。
我们的数据提供了 PLS 潜在的新分子机制。溶酶体功能不足引起的自噬受损可能是 PLS 的一个新的治疗靶点。