Barison Ilaria, Perazzolo Diego, Castellani Chiara, Giarraputo Alessia, Rossi Elisabetta, Vedovelli Luca, Minuzzo Sonia Anna, Tessari Chiara, Pradegan Nicola, Toscano Giuseppe, Tona Francesco, Basso Cristina, Gerosa Gino, Mandruzzato Susanna, Abate Davide, Gregori Dario, Angelini Annalisa, Fedrigo Marny
Cardiovascular Pathology, Department of Cardiac, Thoracic and Vascular Sciences and Public Health, University of Padova, Padova, Italy.
Department of Surgery, Oncology and Gastroenterology, Oncology Section, University of Padova, Padova, Italy.
Front Immunol. 2025 May 16;16:1581151. doi: 10.3389/fimmu.2025.1581151. eCollection 2025.
Cytomegalovirus (CMV) infection is a relevant threat to heart-transplanted patients during the first year after surgery, leading to increased morbidity and, in some cases, mortality. This study aims to assess the transcriptomic profile of CMV infection in cardiac transplanted patients as a new diagnostic approach to discriminate infection and Acute Cellular Rejection (ACR) on EMB specimens.
We performed a microarray-based messenger RNA (mRNA) and micro-RNA (miRNA) profiling. We analyzed three patient groups in the setting of CMV viremia and inflammatory infiltrate: a control group (n=5), an ACR group (n=5), and an infection group (n=6). Differentially expressed mRNA and miRNA were further investigated through bioinformatic pathway analysis.
Focusing on infection vs rejection comparison, we investigated the role of the 18 differentially expressed mRNAs and the 12 miRNAs with the most significative p-value (gene level fold change, FC <-2 or >2, p-value <0.05). Based on the bioinformatic analysis, we explored the regulatory effects of these miRNAs on the mRNA pathways independently identified in the same samples. The results showed that two genes, IL7R and GZMK (-38.63 and -3.15 FC, respectively), and two miRNAs, mir-93-5p and mir-345-5p (-2.63 and -2.18 FC, respectively), are differentially expressed in infection and can be exploited to differentiate CMV-positive from ACR-positive EMB specimens, reaching an AUC of 0.87 and an accuracy of 91% at cross-validation.
We have identified a distinctive combined molecular profile of mRNAs and miRNAs for infection in post-cardiac transplant follow-up. Based on IL7R, GZMK, mir-93-5p, and mir-345-5p we suggest a novel possible workflow to distinguish infection, where those markers are downregulated, from rejection, where they are overexpressed, on EMB specimens. This analysis showed good accuracy and promising predictive performance. The future combined analysis of these genes and these miRNAs through user-friendly techniques, such as quantitative PCR, could reduce turn-around time and improve our diagnostic power for distinguishing CMV infection from ACR in EMB specimens.
巨细胞病毒（CMV）感染是心脏移植患者术后第一年面临的一个重要威胁，会导致发病率增加，在某些情况下还会导致死亡。本研究旨在评估心脏移植患者CMV感染的转录组特征，作为一种新的诊断方法，用于区分心内膜活检（EMB）标本中的感染和急性细胞排斥反应（ACR）。
我们进行了基于微阵列的信使核糖核酸（mRNA）和微小核糖核酸（miRNA）分析。我们分析了三组处于CMV病毒血症和炎性浸润状态的患者：一个对照组（n = 5）、一个ACR组（n = 5）和一个感染组（n = 6）。通过生物信息通路分析进一步研究差异表达的mRNA和miRNA。
聚焦于感染与排斥的比较，我们研究了18个差异表达的mRNA和12个p值最显著的miRNA（基因水平的倍数变化，FC < -2或> 2，p值< 0.05）的作用。基于生物信息分析，我们探索了这些miRNA对在同一样本中独立鉴定出的mRNA通路的调控作用。结果显示，两个基因，IL7R和GZMK（FC分别为-38.63和-3.15），以及两个miRNA，mir-93-5p和mir-345-5p（FC分别为-2.63和-2.18），在感染中差异表达，可用于区分CMV阳性和ACR阳性的EMB标本，交叉验证时曲线下面积（AUC）为0.87，准确率为91%。
我们已经确定了心脏移植术后随访中感染的一种独特的mRNA和miRNA联合分子特征。基于IL7R、GZMK、mir-93-5p和mir-345-5p，我们提出了一种新的可能的工作流程，用于区分EMB标本中感染（这些标志物下调）和排斥（这些标志物上调）。该分析显示出良好的准确性和有前景的预测性能。未来通过诸如定量PCR等用户友好技术对这些基因和这些miRNA进行联合分析，可能会减少周转时间，并提高我们在EMB标本中区分CMV感染和ACR的诊断能力。
