组织学上，黑色素瘤组织中与焱死亡相关基因 (PRGs)、GZMA、GSDMB、NLRP1、IL18和CHMP4A阳性细胞的百分比较正常皮肤有所减少。焱死亡是癌症领域的新前沿，对肿瘤微环境和肿瘤免疫治疗有影响。然而，焱死亡的作用尚存在争议，部分原因是由于黑色素瘤中细胞组成的异质性。在本研究中，我们对黑色素瘤标本中焱死亡的单细胞转录组景观进行了全面分析。我们的研究结果显示，PRGs的表达失调，尤其是在免疫细胞中，如CD8+细胞（代表CD8+ T细胞）和CD57+细胞（代表NK细胞）。此外，免疫组织化学和多重免疫荧光染色实验证实了GZMA+细胞和GSDMB+细胞主要在免疫细胞中表达，特别是在CD8+ T细胞和NK细胞中。黑色素瘤标本中，GZMA+融合CD8+ T细胞的存在率为0.11%，GSDMB+融合CD57+细胞的存在率为0.08%，相比之下，对照组的存在率分别为4.02%和0.62%。上述研究结果表明，肿瘤内免疫细胞的减少可能在减弱焱死亡能力方面起作用，从而在抗黑色素瘤特性方面存在潜在风险。为了量化临床相关性，我们构建了一个预后风险模型和一个个体化直观图（C-index=0.58，P=0.002），提示PRGs在恶性黑色素瘤预防中可能发挥作用。总之，我们的综合单细胞和批量RNA测序分析鉴定了免疫细胞集群和免疫基因模块，并进行了实验证实，有助于更好地理解黑色素瘤中的焱死亡。 © 2023. 作者。

Histologically, melanoma tissues had fewer positive cells percentage of pyroptosis-related genes (PRGs), GZMA, GSDMB, NLRP1, IL18, and CHMP4A in epidermal than in normal skin. Pyroptosis, a new frontier in cancer, affects the tumor microenvironment and tumor immunotherapy. Nevertheless, the role of pyroptosis remains controversial, which reason is partly due to the heterogeneity of the cellular composition in melanoma. In this study, we present a comprehensive analysis of the single-cell transcriptome landscape of pyroptosis in melanoma specimens. Our findings reveal dysregulation in the expression of PRGs, particularly in immune cells, such as CD8+ cells (representing CD8+ T cells) and CD57+ cells (representing NK cells). Additionally, the immunohistochemical and multiplex immunofluorescence staining experiments results further confirmed GZMA+ cells and GSDMB+ cells were predominantly expressed in immune cells, especially in CD8 + T cells and NK cells. Melanoma specimens secreted a minimal presence of GZMA+ merged CD8+ T cells (0.11%) and GSDMB+ merged CD57+ cells (0.08%), compared to the control groups exhibiting proportions of 4.02% and 0.62%, respectively. The aforementioned findings indicate that a reduced presence of immune cells within tumors may play a role in diminishing the ability of pyroptosis, consequently posing a potential risk to the anti-melanoma properties. To quantify clinical relevance, we constructed a prognostic risk model and an individualized nomogram (C-index=0.58, P = 0.002), suggesting a potential role of PRGs in malignant melanoma prevention. In conclusion, our integrated single-cell and bulk RNA-seq analysis identified immune cell clusters and immune gene modules with experiment validation, contributing to our better understanding of pyroptosis in melanoma.© 2023. The Author(s).