Liang Xiaoyan, Schnaper H William, Matsusaka Taiji, Pastan Ira, Ledbetter Steve, Hayashida Tomoko
Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, United States of America.
Department of Internal Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
PLoS One. 2016 May 17;11(5):e0155534. doi: 10.1371/journal.pone.0155534. eCollection 2016.
Fibrosis is a final common pathway leading to loss of kidney function, in which the fibrogenic cytokine, transforming growth factor β (TGF-β), plays a central role. While previous studies showed that TGF-β antagonism by various means prevents fibrosis in mouse models, clinical approaches based on these findings remain elusive. 1D11 is a neutralizing antibody to all three isoforms of TGF-β. In both adriamycin (ADR)-induced nephropathy and NEP25 podocyte ablation nephropathy, thrice-weekly intraperitoneal administration of 1D11 from the day of disease induction until the mice were sacrificed (day 14 for ADR and day 28 for NEP25), significantly reduced glomerular COL1A2 mRNA accumulation and histological changes. Consistent with our previous findings, proteinuria remained overt in the mice treated with 1D11, suggesting distinct mechanisms for proteinuria and fibrogenesis. Podocyte numbers determined by WT1 staining were significantly reduced in NEP25-model glomeruli as expected, while WT1-positive cells were preserved in mice receiving 1D11. Even when 1D11 was administered after the onset of proteinuria on day 3, 1D11 preserved WT1-positive cell numbers in glomeruli and significantly reduced glomerular scar score (2.5 ± 0.2 [control IgG] vs. 1.8 ± 0.2 [1D11], P < 0.05) and glomerular COL1A2 mRNA expression (19.3 ± 4.4 [control IgG] vs. 8.4 ± 2.4 [1D11] fold increase over the healthy control, P < 0.05). Transmission electron microscopy revealed loss of podocytes and denuded glomerular basement membrane in NEP25 mice with disease, whereas podocytes remained attached to the basement membrane, though effaced and swollen, in those receiving 1D11 from day 3. Together, these data suggest that TGF-β neutralization by 1D11 prevents glomerular fibrosis even when started after the onset of proteinuria. While overt proteinuria and podocyte effacement persist, 1D11 prevents total podocytes detachment, which might be a key event activating fibrogenic events in glomeruli.
纤维化是导致肾功能丧失的最终共同途径,其中促纤维化细胞因子转化生长因子β(TGF-β)起着核心作用。虽然先前的研究表明,通过各种手段拮抗TGF-β可在小鼠模型中预防纤维化,但基于这些发现的临床方法仍然难以捉摸。1D11是一种针对TGF-β所有三种亚型的中和抗体。在阿霉素(ADR)诱导的肾病和NEP25足细胞消融性肾病中,从疾病诱导之日起直至小鼠处死(ADR为第14天,NEP25为第28天),每周三次腹腔注射1D11,可显著减少肾小球COL1A2 mRNA积累和组织学变化。与我们之前的研究结果一致,接受1D11治疗的小鼠蛋白尿仍然明显,这表明蛋白尿和纤维化的机制不同。如预期的那样,通过WT1染色确定的NEP25模型肾小球中的足细胞数量显著减少,而在接受1D11的小鼠中WT1阳性细胞得以保留。即使在第3天蛋白尿发作后给予1D11,1D11也能保留肾小球中WT1阳性细胞数量,并显著降低肾小球瘢痕评分(2.5±0.2[对照IgG]对1.8±0.2[1D11],P<0.05)和肾小球COL1A2 mRNA表达(相对于健康对照,19.3±4.4[对照IgG]对8.4±2.4[1D11]倍增加,P<0.05)。透射电子显微镜显示,患病的NEP25小鼠中足细胞丢失且肾小球基底膜裸露,而从第3天开始接受1D11治疗的小鼠中,足细胞虽然扁平且肿胀,但仍附着于基底膜。总之,这些数据表明,即使在蛋白尿发作后开始使用1D11中和TGF-β也可预防肾小球纤维化。虽然明显的蛋白尿和足细胞扁平仍然存在,但1D11可防止足细胞完全脱离,这可能是激活肾小球纤维化事件的关键事件。