Changes in cardiac gene expression after pig-to-primate orthotopic xenotransplantation
Changes in cardiac gene expression after pig-to-primate orthotopic xenotransplantation
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DOI：
10.1111/j.1399-3089.2010.00620.x
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发表时间：
2011-01-01
影响因子：
3.9
通讯作者：
McGregor, Christopher G. A.
中科院分区：
文献类型：
--
作者：
Byrne, Guerard W.;Du, Zeji;McGregor, Christopher G. A.
背景资料：基因谱分析方法已广泛用于描绘基因表达的变化，作为深入了解排斥反应或疾病病理机制的方法。在此，我们使用基因谱来比较与不同的原位心脏异种移植（OCXTx）结果相关的基因表达的变化，并确定OCXTx对心脏生理学的潜在影响。方法：我们使用Affyssin基因芯片猪基因组阵列来表征三种类型的原位心脏异种移植结果：1）经历延迟异种移植排斥（DXR）的排斥心脏; 2）存活心脏，其中异种移植物未被排斥，受体死亡是由于模型并发症;和3）在移植的前48小时内未能提供足够的循环支持的心脏，称为“围手术期心脏异种移植物功能障碍”（PCXD）。将每组中的基因表达与对照、未移植的猪心脏进行比较，并分析来自对照样品的> 3个标准偏差（+/-3SD）的基因表达的变化。生物信息学分析用于鉴定参与京都基因和基因组途径百科全书和基因个体发育分子功能的基因的富集。定量RT-PCR.Results：+/- 3SD数据集包含260个探针，与对照猪心脏相比，基因表达的变化最小为3.5倍。分层聚类分析分离拒绝，幸存者和PCXD样品，表明每个组的基因表达的独特变化。所有移植结果共有一组21个探针，具有相似的表达改变，这表明正在进行的心肌炎症和损伤。一些结果特异性的基因表达的变化进行了鉴定。生物信息学分析检测到丰富的基因参与蛋白质，碳水化合物和支链氨基酸代谢，细胞外基质受体相互作用，粘着斑，和细胞communication.Conclusions：这是第一个全基因组的评估后，OCXTx心脏基因表达的变化。分层聚类分析表明每个移植结果的独特基因谱，但需要额外的样品来定义独特的分类器探针集。定量RT-PCR证实，所有移植物均表现出与细胞因子和血管抗体介导的炎症作用一致的持续炎症和心肌损伤的强有力证据。这也与生物信息学分析一致，表明幸存者和PCXD样品中正在进行组织修复。生物信息学分析首次表明，异种移植可能会影响心脏代谢的幸存者和拒绝的样品。这项研究强调了分子分析的潜在效用，以监测异种移植功能，以确定新的分子标记物和了解过程，这可能有助于DXR。
Background: Gene profiling methods have been widely useful for delineating changes in gene expression as an approach for gaining insight into the mechanism of rejection or disease pathology. Herein, we use gene profiling to compare changes in gene expression associated with different orthotopic cardiac xenotransplantation (OCXTx) outcomes and to identify potential effects of OCXTx on cardiac physiology.Methods: We used the Affymetrix GeneChip Porcine Genomic Array to characterize three types of orthotopic cardiac xenograft outcomes: 1) rejected hearts that underwent delayed xenograft rejection (DXR); 2) survivor hearts in which the xenograft was not rejected and recipient death was due to model complications; and 3) hearts which failed to provide sufficient circulatory support within the first 48 h of transplant, termed "perioperative cardiac xenograft dysfunction" (PCXD). Gene expression in each group was compared to control, not transplanted pig hearts, and changes in gene expression > 3 standard deviations (+/- 3SD) from the control samples were analyzed. A bioinformatics analysis was used to identify enrichments in genes involved in Kyoto Encyclopedia of Genes and Genomes pathways and gene ontogeny molecular functions. Changes in gene expression were confirmed by quantitative RT-PCR.Results: The +/- 3SD data set contained 260 probes, which minimally exhibited a 3.5-fold change in gene expression compared to control pig hearts. Hierarchical cluster analysis segregated rejected, survivor and PCXD samples, indicating a unique change in gene expression for each group. All transplant outcomes shared a set of 21 probes with similarly altered expression, which were indicative of ongoing myocardial inflammation and injury. Some outcome-specific changes in gene expression were identified. Bioinformatics analysis detected an enrichment of genes involved in protein, carbohydrate and branched amino acid metabolism, extracellular matrix-receptor interactions, focal adhesion, and cell communication.Conclusions: This is the first genome wide assessment of changes in cardiac gene expression after OCXTx. Hierarchical cluster analysis indicates a unique gene profile for each transplant outcome but additional samples will be required to define the unique classifier probe sets. Quantitative RT-PCR confirmed that all transplants exhibited strong evidence of ongoing inflammation and myocardial injury consistent with the effects of cytokines and vascular antibody-mediated inflammation. This was also consistent with bioinformatic analysis suggesting ongoing tissue repair in survivor and PCXD samples. Bioinformatics analysis suggests for the first time that xenotransplantation may affect cardiac metabolism in survivor and rejected samples. This study highlights the potential utility of molecular analysis to monitor xenograft function, to identify new molecular markers and to understand processes, which may contribute to DXR.