McGregor Hugh, Sun Zhengda, McCoy David, Kumar Vishal, Conrad Miles, Wilson Mark, Cooke Daniel
Department of Radiology and Biomedical Imaging, University of California, 505 Parnassus Avenue, M-391, San Francisco, California 94143-0628.
Department of Radiology and Biomedical Imaging, University of California, 505 Parnassus Avenue, M-391, San Francisco, California 94143-0628.
J Vasc Interv Radiol. 2018 Oct;29(10):1403-1409.e2. doi: 10.1016/j.jvir.2018.04.034. Epub 2018 Aug 31.
To demonstrate feasibility of endothelial cell (EC) biopsy from dialysis arteriovenous fistulas (AVFs) with the use of guidewires and to characterize gene expression differences between ECs from stenotic and nonstenotic outflow vein segments.
Nine consecutive patients undergoing fistulography for AVF dysfunction from June to August 2016 were enrolled. ECs were biopsied with the use of guidewires from venous outflow stenoses and control outflow veins central to the stenoses. ECs were sorted with the use of flow cytometry, and the Fluidigm Biomark HD system was used for single-cell quantitative polymerase chain reaction (qPCR) analysis of gene expression. Forty-eight genes were assessed and were selected based on different cellular functions and previous literature. Linear mixed models (LMMs) were used to identify differential gene expression between the groups, and self-organizing maps (SOMs) were used to identify cell clusters based on gene coexpression profiles.
A total of 219 and 213 ECs were sampled from venous outflow stenoses and control vein segments, respectively. There were no immediate biopsy-related complications. Forty-eight cells per patient were sorted for qPCR analysis. LMM identified 7 genes with different levels of expression at stenotic segments (P < .05), including AGTR-2, HMOX-2, MTHFR, SERPINC-1, SERPINE-1, SMAD-4, and VWF. SOM analysis identified 4 cell clusters with unique gene expression profiles, each containing stenotic and control ECs.
EC biopsy from dialysis AVFs with the use of guidewires is feasible. Gene expression data suggest that genes involved in multiple cellular functions are dysregulated in stenotic areas. SOMs identified 4 unique clusters of cells, indicating EC phenotypic heterogeneity in outflow veins.
利用导丝证明从透析动静脉内瘘（AVF）获取内皮细胞（EC）活检样本的可行性，并描述狭窄和非狭窄流出静脉段的EC之间的基因表达差异。
纳入2016年6月至8月连续9例因AVF功能障碍接受瘘管造影的患者。使用导丝从静脉流出狭窄处及狭窄中央的对照流出静脉获取EC活检样本。使用流式细胞术对EC进行分选，并使用Fluidigm Biomark HD系统对基因表达进行单细胞定量聚合酶链反应（qPCR）分析。评估了48个基因，这些基因基于不同的细胞功能和既往文献进行选择。使用线性混合模型（LMM）识别两组之间的差异基因表达，并使用自组织映射（SOM）根据基因共表达谱识别细胞簇。
分别从静脉流出狭窄处和对照静脉段采集了219个和213个EC样本。未发生与活检直接相关的并发症。对每位患者48个细胞进行分选以进行qPCR分析。LMM识别出7个在狭窄段表达水平不同的基因（P < 0.05），包括AGTR-2、HMOX-2、MTHFR、SERPINC-1、SERPINE-1、SMAD-4和VWF。SOM分析识别出4个具有独特基因表达谱的细胞簇，每个簇均包含狭窄处和对照EC。
利用导丝从透析AVF获取EC活检样本是可行的。基因表达数据表明，参与多种细胞功能的基因在狭窄区域失调。SOM识别出4个独特的细胞簇，表明流出静脉中EC存在表型异质性。
