Division of Vascular Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510800, China.
National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Disease, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, China.
Mol Biol Rep. 2024 Nov 7;51(1):1127. doi: 10.1007/s11033-024-10050-5.
Developing a molecular signature associated with CD4 + T cell infiltration is essential for identifying biomarkers in abdominal aortic aneurysms (AAA). Establishing such a signature is vital for improving diagnostic accuracy and therapeutic strategies for AAA. This study focuses on CD4 + T cells, which are pivotal in the immune microenvironment of AAA, to pinpoint key targets.
We identified CD4 + T cell-related biomarkers in AAA using bulk and single-cell RNA sequencing data from the GEO database. We employed CIBERSORT to assess immune cell infiltration and applied weighted gene co-expression network analysis and differential expression analyses to pinpoint key genes. A nomogram and related score were developed based on these genes. Single-cell RNA sequencing further analyzed their expression across cell types, and KEGG/GO analyses were conducted for candidate genes. Four genes (NUPR1, CCL4L2, CCL3L3, MMP9) were identified finally. Validation via qPCR and immunohistochemistry showed NUPR1 downregulation in aneurysms and an inverse relationship with CD4 + T cells infiltration. Immunofluorescence results indicated NUPR1 was mainly expressed in the cytoplasm of vascular smooth muscle cells (VSMCs). After VSMCs-specific overexpression of NUPR1 via adeno-associated virus, the AAA diameter decreased, while treatment with the NUPR1 nuclear translocation inhibitor ZZW-115 hydrochloride had no effect on AAA size. Overexpression of NUPR1 in VSMCs suppresses the migration of CD4 + T cells.
The four-gene signature accurately predicts CD4 + T cell infiltration in AAA patients and may serve as a clinical index. NUPR1 could be a therapeutic target for the interaction between CD4 + T cells and AAA.
开发与 CD4+T 细胞浸润相关的分子特征对于鉴定腹主动脉瘤 (AAA) 的生物标志物至关重要。建立这样的特征对于提高 AAA 的诊断准确性和治疗策略至关重要。本研究聚焦于 CD4+T 细胞，它们在 AAA 的免疫微环境中起着关键作用，以确定关键靶点。
我们使用 GEO 数据库中的批量和单细胞 RNA 测序数据鉴定了 AAA 中与 CD4+T 细胞相关的生物标志物。我们使用 CIBERSORT 评估免疫细胞浸润，并应用加权基因共表达网络分析和差异表达分析来确定关键基因。基于这些基因开发了列线图和相关评分。单细胞 RNA 测序进一步分析了它们在细胞类型中的表达，对候选基因进行了 KEGG/GO 分析。最终确定了四个基因 (NUPR1、CCL4L2、CCL3L3、MMP9)。通过 qPCR 和免疫组织化学验证显示 NUPR1 在动脉瘤中下调，与 CD4+T 细胞浸润呈负相关。免疫荧光结果表明 NUPR1 主要表达在血管平滑肌细胞 (VSMCs) 的细胞质中。通过腺相关病毒特异性过表达 NUPR1 后，AAA 直径减小，而 NUPR1 核转位抑制剂 ZZW-115 盐酸盐处理对 AAA 大小没有影响。VSMCs 中 NUPR1 的过表达抑制 CD4+T 细胞的迁移。
该四基因特征准确预测 AAA 患者中 CD4+T 细胞浸润，可能作为临床指标。NUPR1 可能是 CD4+T 细胞与 AAA 相互作用的治疗靶点。
