Single cell RNA sequencing of paw skin from healthy and Col7a1 knockout (RDEB) mice. Single cell RNA sequencing of paw skin from healthy and Col7a1 knockout (RDEB) mice

Recessive dystrophic epidermolysis bullosa (RDEB) is a genetic disorder caused by loss of function of the Col7a1 gene that encodes collagen VII, a critical structural protein that anchors the epidermis to the dermis. Manifestations of this disease include chronic wounding, blistering, immune dysfunction, and eventually squamous cell carcinoma. Symptoms are likely due to the disruption and dysregulation of the dermal microenvironment. Mice with RDEB symptoms, due to Col7a1 knockout, have been developed to study the disease's development and pathological manifestations in an in vivo model. Single cell RNA sequencing was performed on a single cell suspension of the paw skin of 2 week old RDEB mice, along with their wild-type littermates, to create a transcriptomic view of their dermal microenvironment. We analyzed the sequencing data at different resolutions. One focusing on the overall tissue level as well as closer analysis of specific cell types (fibroblasts, keratinocytes, immune cells, and vascular cells). Overall design: Single cell supsensions were extracted from the paw skin of four, 2 week old mice. Two mice had Col7a1 knocked-out and displayed symptoms characteristic of RDEB, while the other two were normal.

隐性遗传性营养不良性大疱性表皮松解症（RDEB）是一类遗传性疾病，由编码Ⅶ型胶原（collagen VII）的Col7a1基因功能缺失引发；Ⅶ型胶原是将表皮锚定至真皮的关键结构蛋白。该疾病的临床表现包括慢性创面形成、水疱生成、免疫功能异常，最终可进展为鳞状细胞癌（squamous cell carcinoma）。其症状大概率源于真皮微环境的破坏与调控失衡。科研人员已构建Col7a1基因敲除的RDEB症状小鼠模型，用于在活体模型中研究该病的发病进程与病理表现。本研究针对2周龄RDEB小鼠及其野生型同窝幼鼠的爪部皮肤单细胞悬液开展单细胞RNA测序（Single cell RNA sequencing），以解析其真皮微环境的转录组全景。我们以不同分辨率对测序数据进行分析：一方面聚焦整体组织水平，另一方面针对成纤维细胞、角质形成细胞、免疫细胞及血管细胞等特定细胞类型开展精细化分析。实验整体设计如下：从4只2周龄小鼠的爪部皮肤提取单细胞悬液，其中2只为Col7a1基因敲除小鼠，表现出RDEB的典型症状，剩余2只为野生型正常小鼠。