瘢痕疙瘩放疗前后关键细胞亚群的变化及相互作用  

The changes and interactions of key cell subpopulations in keloids before and after radiotherapy

作　　者：李伟[1] 凃贝霖 李晓倩 许学文[1] 肖海涛[1] 张艳阁[1] 张舒羽 Li Wei;Tu Beilin;Li Xiaoqian;Xu Xuewen;Xiao Haitao;Zhang Yange;Zhang Shuyu(Department of Plastic and Burn Surgery,West China Hospital,Sichuan University,Chengdu 610041,China;West China School of Basic Medical Sciences&Forensic Medicine,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院整形外科/烧伤科,成都610041 [2]四川大学华西基础与法医学院,成都610041

出　　处：《中华放射医学与防护杂志》2024年第11期917-923,共7页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金(82073477);四川省科技计划项目(2022JDJQ0051,2023NSFSC0648)。

摘　　要：目的采用单细胞转录组测序技术探索瘢痕疙瘩放疗前后细胞及胞内基因表达,解析放疗前后瘢痕疙瘩及正常皮肤组织细胞间的异质性,识别放疗前后细胞亚群、差异表达基因以及细胞相互作用的变化。方法本研究纳入了4例患者共12个样本,收集每例患者放疗前、后的瘢痕疙瘩及其周围未放疗的正常皮肤组织样本各1个。胸部瘢痕疙瘩从中线处分为左右两侧,分别采取不同放疗方案。左侧采用传统放疗方案,即瘢痕疙瘩切除术后当天开始连续电子线照射4 d(单次5 Gy,共20 Gy)。右侧放疗方案为术前2 d(单次5 Gy,共10 Gy),术后2 d(单次5 Gy,共10 Gy)。结果共分析了25573个成纤维细胞,分为9个异质性亚群(fibroblasts 1~9)。Ⅱ型成纤维细胞比例在放疗后增加(t=4.70,P<0.05)。放疗后经典单核细胞及巨噬细胞的数目增加,但差异无统计学意义(P>0.05)。进一步将巨噬细胞(4723个细胞)分为4类,细胞通讯分析(CellPhoneDB)提示,Ⅲ型巨噬细胞与各大类成纤维细胞的相互作用明显更为密切(F=64.30,P<0.05),主要涉及Collagen及Chemerin信号通路,相互作用在放疗后的瘢痕疙瘩样本中更为突出(P<0.05)。结论Ⅲ型巨噬细胞与成纤维细胞的相互作用可能是未来瘢痕疙瘩放疗增敏研究的一个重要切入点。ObjectiveTo explore the heterogeneity among keloids before and after radiotherapy and identify the changes of key cell subpopulations and their interactions utilizing single cell RNA sequencing technology.MethodsFour patients provided a total of 12 samples,each consisting of keloid tissue before and after radiotherapy and the normal skin tissue adjacent to the untreated keloid.The keloid was divided into left and right sides from the midline,and the left-side keloid was fractionally irradiated with 20 Gy electron beam in total in 4 consecutive days.The right-side keloid was irradiated with 10 Gy in 2 fractions before surgery and 10 Gy in 2 fractions after surgery.ResultsA total of 25573 fibroblasts were analyzed and categorized into nine subgroups(fibroblasts 1-9).The proportion of fibroblast-2 increased after radiotherapy(t=4.70,P<0.05).The number of classical monocytes and macrophages increased after radiotherapy,but there was no significant difference due to the shorter time of sample taking at 2 d after radiotherapy(P>0.05).Macrophages(4723 cells)were further divided into four categories.CellPhoneDB analysis showed that type-3 macrophages interacted significantly more closely with fibroblasts than type-1 and type-2 macrophages.The most prominent signaling pathways for the interactions between type-3 macrophages and major fibroblast subtypes were the collagen signaling pathway and the chemerin signaling pathway.These interactions were more pronounced in the keloid samples after radiotherapy.ConclusionsThe interactions between type-3 macrophages and fibroblasts(such as fibroblast-2)may serve as an important point for future studies on radio-sensitization of keloids.

关 键 词：单细胞转录组测序 瘢痕疙瘩 放疗 成纤维细胞 巨噬细胞 

分 类 号：R622[医药卫生—整形外科] R815[医药卫生—外科学]