抗TNF治疗改变JIA患者PBMC基因表达谱，可预测疗效

Moorthy LN, et al. ACR 2007. Presentation No:1713.

背景：我们假设儿童期发生的特发性关节炎(JIA)和SLE的基因表达谱是独特的，抗细胞因子或细胞毒药物将改变之，并可能有预测疗效的价值。

目的：利用核酸微阵列技术，分析Etanercept治疗JIA以及环孢霉素/Rituximab(COME)联合治疗SLE，对患者外周血单个核细胞(PBMC)基因表达谱的影响。

方法：共 有4例活动性JIA[3例多关节型(PoJIA)，1例系统型(SoJIA)]以及1例活动性SLE，生物制剂治疗前和治疗后(90±30天)采集患者的 外周血。另设2例年龄匹配患者作为对照(1例SoJIA，1例链球菌感染后反应性关节炎)，疾病无活动，也不接受任何治疗。除了分析治疗前后配对数据，还 比较治疗有效患者(3例PoJIA中的2例)与无效者(1例PoJIA，1例SoJIA)以及1例SLE。

结果：PoJIA、 SoJIA、SLE以及对照组患者均有独特的基因表达印章。共发现157个差异表达基因。Etanercept治疗后，80个基因下调，77个基因上调 (p=0.01-0.0001)。3例PoJIA治疗后，参与炎症通路的以及已知受TNF调控的基因出现下调，这与体内TNF受到功能性抑制相一致。治疗 后上调基因有：UBE2V1，参与NFkB途径；CD74，MHC-II偶联的不变链；HSP90AB1，PKR的一种调控子，是I型干扰素途径中的成 员；TLN1，维系正常整合素的功能。SLE中过表达的某些基因也见于PoJIA治疗无效者(CALR、IFNG、STAT1)以及 SoJIA(SLC16A3、MMP9、VSIG4、DEFA1和4、ARG1、CYP453、CEACAM8和6、ANXA3、OLFM4)。

结论：初步分析提示Etanercept可以改变JIA患者PBMC的基因表达谱，基因表达特征可以区分出治疗有效与无效者。JIA患者中如有一种狼疮样的表达印章可能预示抗TNF治疗疗效不佳。

原文文摘如下。

Effect on Anti-TNF Agents on Gene Expression in Children with Juvenile Arthritis

Present ID: 1713.

We hypothesize that gene expression patterns for childhood juvenile idiopathic arthritis (JIA) and systemic lupus erythematosus (SLE) are unique, will be altered by anti-cytokine/cytotoxic agents, and may predict response to therapy.
Purpose: To examine the effect of etanercept in JIA and of cyclophosphamide/rituximab combination (COMB) in SLE on PBMC gene expression using microarray-based methods.
Methods: Paired blood samples were collected from 4 children with active JIA [3 polyarticular (PoJIA), 1 systemic onset (SoJIA)] and one with active SLE prior to and 90±30 days after initiation of therapy with etanercept for the JIA patients and COMB therapy for the SLE patient. Paired blood samples from 2 age-matched controls (1 SoJIA, 1 post-streptococcal reactive arthritis) with inactive disease on no therapy were also collected. PBMCs were separated, total RNA isolated and microarray experiments were conducted using Affymetrix chips (HGU133 plus 2). Pre and post treatment data were compared. The t-test (cut-off alpha 0.01), fold change tests and change expression analysis using Affymetrix MAS software were performed to compare the 2 groups. Change analysis compares the pre and post treatment samples from the same patient and is done on the level of probes (as opposed to probe-sets). In addition, data from responders (2 of 3 PoJIA) and non-responders (1 PoJIA, the SoJIA) and the SLE patient were compared.
Results: PoJIA, SoJIA, SLE and control subjects had distinct gene expression signatures. Probes not scored as present in at least 3 of 4 JIA samples at either time point were removed; 157 differentially expressed genes remained. After etanercept, 80 genes were downregulated and 77 upregulated (p=0.01-0.0001). Genes involved in inflammatory pathways and known to be regulated by TNF (CEBPD, SOCS3, and PBEF1) were decreased after treatment in 3 PoJIA patients, consistent with functional inhibition of TNF in vivo. UBE2V1, involved in the NFkB pathway; CD74, the MHC class II-associated invariant chain; HSP90AB1, a regulator of PKR, a component of the type I interferon pathway; and TLN1, a contributor to normal integrin function, were increased after treatment. Some genes over expressed in the SLE patient were also detected in the non-responder with PoJIA (CALR, IFNG, STAT1) and the non-responder with SoJIA (SLC16A3, MMP9, VSIG4, DEFA1 and 4, ARG1, CYP453, CEACAM8 and 6, ANXA3, OLFM4). Many of these are components or targets of pro-inflammatory cytokine pathways.
Conclusion: Our preliminary analysis suggests that gene expression is modified by etanercept in JIA patients and separates responders from non-responders. A lupus-like signature in JIA patients may predict lack of response to anti-TNF therapy.

L.N. Moorthy, Arthritis Foundation Investigator Award 2007-2009; G. Schemmann, NIH support on colon cancer-related work (unrelated), 2; UMDNJ, Princeton University in Mechanical Engineering Dept, 3; M.K. Crow, NIH, Alliance for Lupus Research, 2; Hospital For Special Surgery, 3; E. Zachariah, None; M. Peterson, Co-investigator on about 3 NIH grants, 2; Hospital For Special Surgery, 3; Dr. Moorthy's Pfizer clinical scholars grant on QOL and SLE; Consultant on 2 NIH grants; Consultant Burke Hospital, 5; Consultant on this project (no- fees), 9; T. Lehman, Hospital For Special Surgery, 3; Genentech, Abbott, Wyeth, 8; D. Notterman, NIH (NCI) support, 2.