Enhanced podocyte differentiation and changing drug toxicity sensitivity through pressure-controlled mechanical filtration stress on a glomerulus-on-a-chip
Enhanced podocyte differentiation and changing drug toxicity sensitivity through pressure-controlled mechanical filtration stress on a glomerulus-on-a-chip
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DOI：
10.1039/d2lc00941b
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发表时间：
2022-12-06
期刊：
影响因子：
6.1
通讯作者：
Fujii, Teruo
中科院分区：
文献类型：
--
作者：
Doi, Kotaro;Kimura, Hiroshi;Fujii, Teruo
足细胞，定位于肾小球，是肾病蛋白尿的预后因素，并暴露于不同的生理刺激，从基础到根尖滤过流。在肾小球疾病的药物发现和疾病建模方面的研究开发了肾小球芯片，研究足细胞力学生物学，实现了动物实验的替代方法。然而，滤过刺激对足细胞的影响尚不清楚。在此，我们报告了一种用户友好的过滤培养设备和系统的成功开发，该设备和系统可以通过结合市售的培养插入物来精确控制空气压力控制过滤流量。它允许小鼠足细胞在过滤条件下培养三天，并保证保持足细胞层的完整性。利用我们的系统，本研究成功地再现了由肾小球高血压引起的高滤过引起的足细胞损伤，这是许多肾小球疾病的常见病理生理，并且从形态和基因表达方面来看，滤过刺激促进了足细胞的成熟。此外，我们证明了过滤刺激诱导足细胞的药物反应性与静态条件下不同，并且药物反应性的差异取决于药理机制。总的来说，本研究揭示了滤过刺激的分化和药效学特性，为实现芯片肾小球的足细胞力学生物学研究领域提供了新的见解。
Podocytes, localized in the glomerulus, are a prognostic factor of proteinuria in kidney disease and are exposed to distinct physiological stimuli from basal to apical filtration flow. Research studies on drug discovery and disease modeling for glomerulopathy have developed a glomerulus-on-a-chip and studied podocyte mechanobiology to realize alternative methods to animal experiments. However, the effect of filtration stimulus on podocytes has remained unclear. Herein, we report the successful development of a user-friendly filtration culture device and system that can precisely control the filtration flow using air pressure control by incorporating a commercially available culture insert. It allows mouse podocytes to be cultured under filtration conditions for three days with a guarantee of maintaining the integrity of the podocyte layer. Using our system, this study demonstrated that podocyte damage caused by hyperfiltration resulting from glomerular hypertension, a common pathophysiology of many glomerulopathies, was successfully recapitulated and that filtration stimulus promotes the maturation of podocytes in terms of their morphology and gene expression. Furthermore, we demonstrated that filtration stimulus induced different drug responsiveness in podocytes than those seen under static conditions, and that the difference in drug responsiveness was dependent on the pharmacological mechanism. Overall, this study has revealed differentiating and pharmacodynamic properties of filtration stimulus and brings new insights into the research field of podocyte mechanobiology towards the realization of glomerulus-on-a-chip.