Nasrallah Ali, Rezvani Hamid-Reza, Kobaisi Farah, Hammoud Ahmad, Rambert Jérôme, Smits Jos P H, Sulpice Eric, Rachidi Walid
Univ. Grenoble Alpes, CEA, Inserm, IRIG, UA13 BGE, Biomics, Grenoble, 38000, France.
Aquiderm, University of Bordeaux, Bordeaux, France.
Sci Rep. 2024 Dec 28;14(1):30879. doi: 10.1038/s41598-024-81675-6.
Xeroderma pigmentosum group C (XPC) is a versatile protein crucial for sensing DNA damage in the global genome nucleotide excision repair (GG-NER) pathway. This pathway is vital for mammalian cells, acting as their essential approach for repairing DNA lesions stemming from interactions with environmental factors, such as exposure to ultraviolet (UV) radiation from the sun. Loss-of-function mutations in the XPC gene confer a photosensitive phenotype in XP-C patients, resulting in the accumulation of unrepaired UV-induced DNA damage. This remarkable increase in DNA damage tends to elevate by 10,000-fold the risk of developing melanoma and non-melanoma skin cancers. To date, creating accurate and reproducible models to study human XP-C disease has been an important challenge. To tackle this, we used CRISPR-Cas9 technology in order to knockout the XPC gene in various human skin cells (keratinocytes, fibroblasts, and melanocytes). After validation of the knockout in these edited skin cells, we showed that they recapitulate the major phenotypes of XPC mutations: photosensitivity and the impairment of UV-induced DNA damage repair. Moreover, these knockout cells demonstrated a reduced proliferative capacity compared to their respective controls. Finally, to better mimic the disease environment, we built a 3D reconstructed skin using these XPC knockout skin cells. This model exhibited an abnormal behavior, showing an extensive remodeling of its extracellular matrix compared to normal skin. Analyzing the composition of the fibroblast secretome revealed a significant augmented shift in the inflammatory response following XPC knockout. Our innovative "disease on a dish" approach can provide valuable insights into the molecular mechanisms underlying XP-C disease, paving the way to design novel preventive and therapeutic strategies to alleviate the disease phenotype. Also, given the high risk of skin cancer onset in XP-C disease, our new approach can serve as a link to draw novel insights into this elusive field.
着色性干皮病C组（XPC）是一种多功能蛋白质，对全球基因组核苷酸切除修复（GG-NER）途径中感知DNA损伤至关重要。该途径对哺乳动物细胞至关重要，是其修复因与环境因素相互作用（如暴露于太阳紫外线（UV）辐射）而产生的DNA损伤的主要方式。XPC基因的功能丧失突变会使XP-C患者出现光敏表型，导致未修复的紫外线诱导的DNA损伤积累。这种DNA损伤的显著增加往往会使患黑色素瘤和非黑色素瘤皮肤癌的风险提高10000倍。迄今为止，创建准确且可重复的模型来研究人类XP-C疾病一直是一项重大挑战。为了解决这个问题，我们使用CRISPR-Cas9技术在各种人类皮肤细胞（角质形成细胞、成纤维细胞和黑素细胞）中敲除XPC基因。在验证这些编辑后的皮肤细胞中的基因敲除后，我们发现它们重现了XPC突变的主要表型：光敏性和紫外线诱导的DNA损伤修复受损。此外，与各自的对照相比，这些基因敲除细胞的增殖能力降低。最后，为了更好地模拟疾病环境，我们使用这些XPC基因敲除的皮肤细胞构建了三维重建皮肤。该模型表现出异常行为，与正常皮肤相比，其细胞外基质发生了广泛重塑。分析成纤维细胞分泌组的组成发现XPC基因敲除后炎症反应有显著增强的变化。我们创新的“培养皿上的疾病”方法可以为XP-C疾病的分子机制提供有价值的见解，为设计减轻疾病表型的新型预防和治疗策略铺平道路。此外，鉴于XP-C疾病中皮肤癌发病风险高，我们的新方法可以作为一个纽带，为这个难以捉摸的领域带来新的见解。
