Abstract：
OBJECTIVE: Calcineurin inhibitors (CNIs) are the backbone for immunosuppression after solid organ transplantation. Although successful in preventing kidney transplant rejection, their nephrotoxic side effects contribute to allograft injury. Renal parenchymal lesions occur for cyclosporine A (CsA) as well as for the currently favored tacrolimus (Tac). We aimed to study whether chronic CsA and Tac exposures, before reaching irreversible nephrotoxic damage, affect renal compartments differentially and whether related pathogenic mechanisms can be identified.
METHODS: CsA and Tac were administered chronically in wild type Wistar rats using osmotic minipumps over 4 weeks. Functional parameters were controlled. Electron microscopy, confocal, and 3D-structured illumination microscopy were used for histopathology. Clinical translatability was tested in human renal biopsies. Standard biochemical, RNA-seq, and proteomic technologies were applied to identify implicated molecular pathways.
RESULTS: Both drugs caused significant albeit differential damage in vasculature and nephron. The glomerular filtration barrier was more affected by Tac than by CsA, showing prominent deteriorations in endothelium and podocytes along with impaired VEGF/VEGFR2 signaling and podocyte-specific gene expression. By contrast, proximal tubule epithelia were more severely affected by CsA than by Tac, revealing lysosomal dysfunction, enhanced apoptosis, impaired proteostasis and oxidative stress. Lesion characteristics were confirmed in human renal biopsies.
CONCLUSIONS: We conclude that pathogenetic alterations in the renal compartments are specific for either treatment. Considering translation to the clinical setting, CNI choice should reflect individual risk factors for renal vasculature and tubular epithelia. As a step in this direction, we share protein signatures identified from multiomics with potential pathognomonic relevance.
METHODS: CsA and Tac were administered chronically in wild type Wistar rats using osmotic minipumps over 4 weeks. Functional parameters were controlled. Electron microscopy, confocal, and 3D-structured illumination microscopy were used for histopathology. Clinical translatability was tested in human renal biopsies. Standard biochemical, RNA-seq, and proteomic technologies were applied to identify implicated molecular pathways.
RESULTS: Both drugs caused significant albeit differential damage in vasculature and nephron. The glomerular filtration barrier was more affected by Tac than by CsA, showing prominent deteriorations in endothelium and podocytes along with impaired VEGF/VEGFR2 signaling and podocyte-specific gene expression. By contrast, proximal tubule epithelia were more severely affected by CsA than by Tac, revealing lysosomal dysfunction, enhanced apoptosis, impaired proteostasis and oxidative stress. Lesion characteristics were confirmed in human renal biopsies.
CONCLUSIONS: We conclude that pathogenetic alterations in the renal compartments are specific for either treatment. Considering translation to the clinical setting, CNI choice should reflect individual risk factors for renal vasculature and tubular epithelia. As a step in this direction, we share protein signatures identified from multiomics with potential pathognomonic relevance.
摘要：
目的：钙调神经磷酸酶抑制剂（CNIs）是实体器官移植后免疫抑制的骨架。尽管成功地预防了肾移植排斥反应，它们的肾毒性副作用导致同种异体移植损伤。对于环孢菌素A（CsA）以及目前偏爱的他克莫司（Tac），都会发生肾实质病变。我们的目的是研究慢性CsA和Tac暴露是否，在达到不可逆的肾毒性损伤之前，不同程度地影响肾脏区室，以及是否可以确定相关的致病机制。
方法:使用渗透微型泵在4周内对野生型Wistar大鼠长期施用CsA和Tac。控制功能参数。电子显微镜,共焦,和3D结构照明显微镜用于组织病理学。在人肾活检中测试了临床可翻译性。标准生化，RNA-seq,和蛋白质组学技术被用来鉴定相关的分子途径。
结果：两种药物均对血管和肾单位造成明显的不同程度的损害。肾小球滤过屏障受Tac影响大于CsA,显示内皮和足细胞的显著恶化以及VEGF/VEGFR2信号传导和足细胞特异性基因表达受损。相比之下,CsA对近端小管上皮的影响比Tac更严重，揭示溶酶体功能障碍，细胞凋亡增强，受损的蛋白稳定和氧化应激。在人肾活检中证实了病变特征。
结论：我们得出的结论是，肾脏小室的病因改变对两种治疗都是特异性的。考虑到临床环境的翻译，CNI的选择应反映肾血管和肾小管上皮细胞的个体危险因素。作为朝这个方向迈出的一步,我们分享了从多组学中鉴定出的具有潜在病理标记相关性的蛋白质特征.
方法:使用渗透微型泵在4周内对野生型Wistar大鼠长期施用CsA和Tac。控制功能参数。电子显微镜,共焦,和3D结构照明显微镜用于组织病理学。在人肾活检中测试了临床可翻译性。标准生化，RNA-seq,和蛋白质组学技术被用来鉴定相关的分子途径。
结果：两种药物均对血管和肾单位造成明显的不同程度的损害。肾小球滤过屏障受Tac影响大于CsA,显示内皮和足细胞的显著恶化以及VEGF/VEGFR2信号传导和足细胞特异性基因表达受损。相比之下,CsA对近端小管上皮的影响比Tac更严重，揭示溶酶体功能障碍，细胞凋亡增强，受损的蛋白稳定和氧化应激。在人肾活检中证实了病变特征。
结论：我们得出的结论是，肾脏小室的病因改变对两种治疗都是特异性的。考虑到临床环境的翻译，CNI的选择应反映肾血管和肾小管上皮细胞的个体危险因素。作为朝这个方向迈出的一步,我们分享了从多组学中鉴定出的具有潜在病理标记相关性的蛋白质特征.
