Laboratory of Molecular and Cell Biology, Istituto Dermopatico dell'Immacolata, IDI-IRCCS, Rome, Italy.
Istituto di Genetica Molecolare CNR, Pavia, Italy.
J Invest Dermatol. 2019 Jan;139(1):38-50. doi: 10.1016/j.jid.2018.06.181. Epub 2018 Sep 25.
Defects in Cockayne syndrome type A (CSA), a gene involved in nucleotide excision repair, cause an autosomal recessive syndrome characterized by growth failure, progressive neurological dysfunction, premature aging, and skin photosensitivity and atrophy. Beyond its role in DNA repair, the CSA protein has additional functions in transcription and oxidative stress response, which are not yet fully elucidated. Here, we investigated the role of CSA protein in primary human keratinocyte senescence. Primary keratinocytes from three patients with CS-A displayed premature aging features, namely premature clonal conversion, high steady-state levels of reactive oxygen species and 8-OH-hydroxyguanine, and senescence-associated secretory phenotype. Stable transduction of CS-A keratinocytes with the wild-type CSA gene restored the normal cellular sensitivity to UV irradiation and normal 8-OH-hydroxyguanine levels. Gene correction was also characterized by proper restoration of keratinocyte clonogenic capacity and expression of clonal conversion key regulators (p16 and p63), decreased NF-κB activity and, in turn, the expression of its targets (NOX1 and MnSOD), and the secretion of senescence-associated secretory phenotype mediators. Overall, the CSA protein plays an important role in protecting cells from senescence by facilitating DNA damage processing, maintaining physiological redox status and keratinocyte clonogenic ability, and reducing the senescence-associated secretory phenotype-mediated inflammatory phenotype.
Cockayne 综合征 A 型(CSA)缺陷是一种核苷酸切除修复相关基因的疾病,导致常染色体隐性遗传综合征,其特征为生长发育迟缓、进行性神经功能障碍、早衰以及皮肤光敏感性和萎缩。CSA 蛋白除了在 DNA 修复中发挥作用外,在转录和氧化应激反应中还有其他功能,但尚未完全阐明。在这里,我们研究了 CSA 蛋白在原代人角质形成细胞衰老中的作用。来自 3 名 CSA-A 患者的原代角质形成细胞表现出过早衰老的特征,即过早的克隆转化、高水平的活性氧和 8-OH-羟基鸟嘌呤以及衰老相关的分泌表型。用野生型 CSA 基因稳定转导 CSA-A 角质形成细胞可恢复正常的细胞对紫外线照射的敏感性和正常的 8-OH-羟基鸟嘌呤水平。基因校正还表现为角质形成细胞集落形成能力的适当恢复和克隆转化关键调节剂(p16 和 p63)的表达、NF-κB 活性的降低,以及其靶标(NOX1 和 MnSOD)的表达和衰老相关的分泌表型介导物的分泌。总之,CSA 蛋白通过促进 DNA 损伤处理、维持生理氧化还原状态和角质形成细胞集落形成能力以及减少衰老相关的分泌表型介导的炎症表型,在保护细胞免于衰老方面发挥重要作用。