基于单细胞测序的新型与 DNA 损伤修复相关基因标志物预测宫颈癌患者生存、免疫浸润和药物敏感性

A novel DNA damage repair-related gene signature predicting survival, immune infiltration and drug sensitivity in cervical cancer based on single cell sequencing.

机构信息

Department of Internal Medicine, Medical College of Hunan Normal University, Changsha, Hunan, China.

The High School Attached to Hunan Normal University, Changsha, China.

出版信息

Front Immunol. 2023 Jun 28;14:1198391. doi: 10.3389/fimmu.2023.1198391. eCollection 2023.

DOI:10.3389/fimmu.2023.1198391

PMID:37449209

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10337997/

Abstract

BACKGROUND

Aberrant DNA damage repair (DDR) is one of the hallmarks of tumors, and therapeutic approaches targeting this feature are gaining increasing attention. This study aims to develop a signature of DDR-related genes to evaluate the prognosis of cervical cancer (CC).

METHODS

Differentially expressed genes were identified between high and low DDR groups of cells from the single-cell RNA sequencing dataset GSE168652 based on DDR scores. Using the ssGSEA and WGCNA methods, DDR-related differentially expressed genes were identified from different patients within the TCGA-CESC cohort. Using Cox analysis and LASSO regression analysis, a DDR-related gene signature was constructed based on the intersection of two groups of differentially expressed genes and DDR-related genes from WGCNA, and validated in GSE52903. Immune cell infiltration analysis, mutation analysis, survival analysis, drug sensitivity analysis, etc., were performed in different groups which were established based on the DDR gene signature scoring. A key gene affecting prognosis was selected and validated through biological experiments such as wound healing, migration, invasion, and comet assays.

RESULTS

A novel DDR-related signature was constructed and the nomogram results showed this signature performed better in predicting prognosis than other clinical features for CC. The high DDR group exhibited poorer prognosis, weaker immune cell infiltration in the immune microenvironment, lower expression of immune checkpoint-related genes, lower gene mutation frequencies and more sensitivity to drugs such as BI.2536, Bleomycin and etc. , , and were expressed at higher levels in CaSki and HeLa cells compared to ECT1 cells. Compared with the native CaSki and HeLa cells, the proliferation, migration, invasion and DDR capabilities of CaSki and HeLa cell lines with suppressed expression were significantly decreased.

CONCLUSION

The 7 DDR-related gene signature was an independent and powerful prognostic biomarker that might effectively evaluate the prognosis of CC and provide supplementary information for a more personalized evaluation and precision therapy. was a potential candidate gene that may affect the DDR capacity of CC cells, and its mechanism of action was worth further in-depth study.

摘要

背景

异常的 DNA 损伤修复（DDR）是肿瘤的标志之一，针对这一特征的治疗方法越来越受到关注。本研究旨在开发 DDR 相关基因特征，以评估宫颈癌（CC）的预后。

方法

基于 DDR 评分，从单细胞 RNA 测序数据集 GSE168652 中鉴定出高和低 DDR 细胞组之间的差异表达基因。使用 ssGSEA 和 WGCNA 方法，从 TCGA-CESC 队列中的不同患者中鉴定 DDR 相关差异表达基因。使用 Cox 分析和 LASSO 回归分析，基于 WGCNA 中两组差异表达基因和 DDR 相关基因的交集构建 DDR 相关基因特征，并在 GSE52903 中进行验证。基于 DDR 基因特征评分，在不同组中进行免疫细胞浸润分析、突变分析、生存分析、药物敏感性分析等。选择并通过划痕愈合、迁移、侵袭和彗星试验等生物学实验验证影响预后的关键基因。

结果

构建了一个新的 DDR 相关特征，列线图结果表明，该特征在预测 CC 预后方面优于其他临床特征。高 DDR 组预后较差，免疫微环境中免疫细胞浸润较弱，免疫检查点相关基因表达较低，基因突变频率较低，对 BI.2536、博来霉素等药物更敏感，，，在 ECT1 细胞中表达水平高于 CaSki 和 HeLa 细胞。与天然 CaSki 和 HeLa 细胞相比，表达下调的 CaSki 和 HeLa 细胞系的增殖、迁移、侵袭和 DDR 能力显著降低。