Integrin β-8, But Not β-5 or-6, Protein Expression Is Increased in Livers of Children With Biliary Atresia
Integrin β-8, But Not β-5 or-6, Protein Expression Is Increased in Livers of Children With Biliary Atresia
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DOI：
10.1097/mpg.0000000000000518
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发表时间：
2014-12-01
影响因子：
2.9
通讯作者：
Nadler, Evan P.
中科院分区：
文献类型：
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作者：
Iordanskaia, Tatiana;Koeck, Emily;Nadler, Evan P.
目的：我们之前的工作通过对胆道闭锁 (BA) 患者的公开数据进行计算机分析，证明整合素 beta-5 和 -8 的信使 RNA 表达发生了改变；然而，由于样本量的原因，我们无法证明蛋白质表达具有统计学上的显着差异。在本研究中，我们重复分析了更多 BA 患者的肝纤维化和整合素蛋白表达，并将其与接受肝活检进行其他诊断的患者进行比较，假设 >= 1 个整合素会出现差异表达。方法：从 2 个合作机构获取肝脏标本。将患有 BA 的婴儿 (n = 23) 的样本与接受新生儿肝炎肝活检的婴儿 (n = 9) 的样本进行比较。所有标本均由 2 位对诊断不知情的病理学家（C.R. 和 R.A.）进行分析。采用标准 Ishak 评分来评估纤维化和炎症，免疫组织化学 (IHC) 阳性分为 0 到 4 级。BA 组和对照组的 IHC 阳性和 Ishak 评分之间的比较使用 Student t 检验进行，由于多重比较，P < 0.01 被认为是显着的。通过组内相关性 (ICC) 评估观察者间变异性。结果：根据 Ishak 评分，BA 患者标本的汇总分析显示纤维化程度明显高于对照组（3.21 +/- 1.82 vs 1.17 +/- 1.00，P < 0.005）。 IHC 评估显示，与对照相比，整合素 α nu β 8 蛋白表达增加（2.67 +/- 0.81 vs 1.72 +/- 0.62，P < 0.005）；然而，整合素α nu beta 5（1.93 +/- 0.84 vs 1.50 +/- 0.90，P = 0.23）或整合素α nu beta 6（0.85 +/- 1.20 vs 0.94 +/- 0.85，P = 0.82）表达没有显着差异。这些数据在个体分析中得到证实。整合素 α nu beta 5 (ICC 0.52) 的观察者间一致性较好，整合素 α nu beta 6 (ICC 0.72) 良好，整合素 α nu beta 8 (ICC 0.79) 和纤维化 (ICC 0.89) 的观察者间一致性非常好。 结论：我们的数据显示，整合素 α nu beta 8，但不是整合素 α nu beta 5 或整合素 α nu beta如图6所示，BA患者的肝脏标本中蛋白质表达增加。这些数据支持越来越多的证据表明转化生长因子-β (TGF-β) 激活是导致 BA 纤维化的原因。抗整合素 alpha nu beta 8 或更多整体整合素阻断策略可能是 BA 的治疗选择，但显然还需要进一步的工作。
Objectives: Our previous work demonstrated altered messenger RNA expression of integrin beta-5 and -8, using an in silico analysis of publically available data from patients with biliary atresia (BA); however, we were unable to demonstrate statistically significant differences in protein expression because of sample size. In the present study, we repeated the analysis of liver fibrosis and protein expression of the integrins in a larger cohort of patients with BA and compared them with patients undergoing liver biopsy for other diagnoses, with the hypothesis that >= 1 of the integrins would be differentially expressed.Methods: Liver specimens were obtained at 2 collaborating institutions. Samples from infants with BA (n = 23) were compared with samples from those who underwent liver biopsy for neonatal hepatitis (n = 9). All of the specimens were analyzed by 2 pathologists (C.R. and R.A.), who were blinded to the diagnoses. Standard Ishak scoring was performed to evaluate fibrosis and inflammation, and immunohistochemical (IHC) positivity was graded from 0 to 4. Comparisons between the IHC positivity and Ishak scoring for the BA and control groups were performed using the Student t test with P < 0.01 considered significant because of the multiple comparisons. Interobserver variability was assessed by intraclass correlation (ICC).Results: Pooled analysis from specimens from patients with BA showed significantly more fibrosis than controls based on Ishak scores (3.21 +/- 1.82 vs 1.17 +/- 1.00, P < 0.005). IHC evaluation showed increased integrin alpha nu beta 8 protein expression when compared with controls (2.67 +/- 0.81 vs 1.72 +/- 0.62, P < 0.005); however, there were no significant differences in integrin alpha nu beta 5 (1.93 +/- 0.84 vs 1.50 +/- 0.90, P = 0.23) or integrin alpha nu beta 6 (0.85 +/- 1.20 vs 0.94 +/- 0.85, P = 0.82) expression. These data were confirmed on individual analysis. Interobserver agreement was fair for integrin alpha nu beta 5 (ICC 0.52), good for integrin alpha nu beta 6 (ICC 0.72), and excellent for integrin alpha nu beta 8 (ICC 0.79) and fibrosis (ICC 0.89).Conclusions: Our data show that integrin alpha nu beta 8, but not integrin alpha nu beta 5 or integrin alpha nu beta 6, protein expression is increased in liver specimens of patients with BA. These data support the mounting evidence that transforming growth factor-beta (TGF-beta) activation is responsible for the fibrosis found in BA. Anti-integrin alpha nu beta 8 or more global integrin blocking strategies may be therapeutic options in BA, but further work is clearly needed.