An advanced ADSC therapy for keloid prevention using identification of functional subgroups by single cell transcriptomic analysis. An advanced ADSC therapy for keloid prevention using identification of functional subgroups by single cell transcriptomic analysis

The clinical treatments for keloids are poor efficacy,There is evidence forADSCs acting as a physiological response to local injury or as rejuvenating mechanisms.But the use of exogenous ADSCs fulfill the general accepted criteria for cell-based therapies, but still need further investigations into their efficiency. In this study, we firstly employed microarray techniques and bioinformatics analysis to screen for whole transcriptome differences involved in keloids, enabling the identification of DEGs and functional molecular associated with keloids mechanism. Secondly, we developed functional clustering of stem cells and marker genes of each cluster through single cell sequencing of ADSCs from healthy adults and pigs by bioinformatics analysis. Finally, we identified ADSC subpopulations with KK and KF therapeutic potential, respectively. anti-fibrotic transcriptomic identification and quantitative clustering was performed, and the function of specific ADSCs cluster was investigated in the hypertrophic scar prophylaxis using the miniature pig model. This study might open the way to a new era of mesenchymal stem cells in the treatment of keloids for promising therapeutic advancements. Overall design: pADSCs were isolated from Bama miniature pigs adipose tissue,and analyzed using scRNAseq.

瘢痕疙瘩（keloids）的临床治疗效果欠佳。现有证据表明，脂肪间充质干细胞（Adipose-derived Mesenchymal Stem Cells, ADSCs）可作为局部损伤的生理应答机制或组织年轻化的调控途径。但外源性ADSCs的应用虽符合细胞治疗的公认标准，其临床疗效仍需进一步研究验证。本研究首先采用微阵列技术与生物信息学分析，筛选瘢痕疙瘩相关的全转录组差异，进而鉴定出与瘢痕疙瘩发病机制相关的差异表达基因（Differentially Expressed Genes, DEGs）及功能分子。其次，本研究通过对健康成人与巴马小型猪来源的ADSCs进行单细胞测序，并结合生物信息学分析，完成了干细胞的功能聚类及各聚类簇的标记基因鉴定。最后，本研究分别鉴定出具备KK与KF治疗潜能的ADSCs亚群。本研究开展了抗纤维化转录组学鉴定与定量聚类分析，并利用小型猪模型探究了特定ADSCs聚类簇在增生性瘢痕预防中的功能。本研究有望为间充质干细胞治疗瘢痕疙瘩的领域开启全新的治疗范式，推动相关治疗的突破性进展。实验整体设计：从巴马小型猪脂肪组织中分离得到猪源脂肪间充质干细胞（porcine ADSCs, pADSCs），并通过单细胞RNA测序（Single-Cell RNA Sequencing, scRNAseq）进行分析。