Unveiling the role of particulate matter (PM) and micro-/nanoplastic (MNP) in glomerular kidney disease with focus on membranous glomerulonephritis (Acronym: UNPLOK)

膜性肾小球肾炎(MGN)是由足细胞抗原结合的自身抗体(AABS)引起的。最常见的是LA2R抗体。然而，目前尚不清楚是什么触发了AAB的产生，以及ABBS如何到达MGN的上皮下间隙，因为对于AABS来说，肾小球滤过屏障(GFB)通常几乎是不通透的。在过去，我们可以证明足细胞和肾小球内皮细胞来源的针对肾连素的microRNAs(一种足细胞产生的基底膜细胞外基质蛋白)可能在MGN中发挥作用。我们假设颗粒物(PM)或微/纳米塑料(MNP)诱导针对GBM分子的MIR，使AABS到达下足细胞间隙并抑制MGN的自噬。这一假设应该在三维肾小球细胞培养模型、转基因斑马鱼模型和小鼠(包括蛋白尿和自噬报告系)、可诱导的足细胞特异性miR过表达系和转基因Pla2r小鼠中进行验证。细胞培养模型以及斑马鱼和小鼠将通过细胞培养液、水和多功能气溶胶浓缩浓缩系统暴露于PM和MNP。在我们的斑马鱼模型中，应该在PM/MNP暴露后研究miR对映体的治疗潜力。样品将使用miR/mRNA阵列、Western Blot、蛋白尿和自噬分析，以及拉曼光谱、纳米GPS技术、扫描电子和离子显微镜(FIB/SEM)和扫描离子电导显微镜(SCICM)等多模式、规模桥接分析的独特组合进行分析。MGN患者的肾脏活检也将包括在内。综合数据将用统计和机器学习方法进行关联和定量评估，并可能预测未来的临床诊断和治疗方案。

Membranous glomerulonephritis (MGN) is caused by autoantibodies (AABs) binding to podocyte antigens. The most frequent AAB is LA2R antibody. However, it remains unknown what triggers AAB production and how ABBs reach the subepithelial space in MGN, because the glomerular filtration barrier (GFB) is normally almost impermeable for AABs. In the past, we could show that podocyte and glomerular endothelial cell derived microRNAs (miRs) that target nephronectin - an extracellular matrix protein of the GBM produced by podocytes - might play a role in MGN. We hypothesize that particulate matter (PM) or Micro-/ nanoplastic (MNP) induce miRs that target GBM molecules enabling AABs to reach the subpodocyte space and inhibit autophagy in MGN. This hypothesis should be tested in three dimensional glomerular cell culture models, transgenic zebrafish models and mice including proteinuria and autophagy reporter lines, inducible podocyte specific miR overexpressing lines and transgenic Pla2r mice. Cell culture models as well as zebrafish and mice will be exposed to PM and MNP via cell culture medium, water and versatile aerosol concentration enrichment system. The therapeutic potential of miR antagomirs should be investigated in our zebrafish model after PM/ MNP exposure. Samples will be analyzed using miR/ mRNA arrays, Western Blot, proteinuria and autophagy assays together with a unique combination of multi-modal, scale-bridging analytics such as Raman spectroscopy, nanoGPS technology, scanning electron- and ion microscopies (FIB/SEM) and scanning ion conductance microscopy (SCICM). Kidney biopsies of patients with MGN will also be included. The comprehensive data will be correlated, quantitatively evaluated with statistical and machine learning methods and might predict future clinical diagnostic and therapeutic options.