Colistin is essential for treating multidrug-resistant Gram-negative bacterial infections but has significant nephrotoxic side effects. Traditional approaches for studying colistin's nephrotoxicity are challenged by the rapid metabolism of its prodrug, colistin methanesulfonate and the difficulty of obtaining adequate plasma from critically ill patients. To address these challenges, we developed the Spheroid Nephrotoxicity Assessing Platform (SNAP), a microfluidic device that efficiently detects colistin-induced toxicity in renal proximal tubular epithelial cell (RPTEC) spheroids within 48 hours using just 200 μL of patient plasma. Our findings demonstrate that SNAP not only promotes higher expression of kidney-specific markers aquaporin-1 (AQP1) and low-density lipoprotein receptor-related protein 2 (LRP2) compared to traditional two-dimensional (2D) cultures, but also exhibits increased sensitivity to colistin, with significant toxicity detected at concentrations of 50 μg ml-1 and above. Notably, SNAP's non-invasive method did not identify nephrotoxicity in plasma from healthy donors, thereby confirming its physiological relevance and showcasing superior sensitivity over 2D cultures, which yielded false-positive results. In clinical validation, SNAP accurately identified patients at risk of colistin-induced nephrotoxicity with 100% accuracy for both early and late onset and demonstrated a 75% accuracy rate in predicting the non-occurrence of nephrotoxicity. These results underline the potential of SNAP in personalized medicine, offering a non-invasive, precise and efficient tool for the assessment of antibiotic-induced nephrotoxicity, thus enhancing the safety and efficacy of treatments against resistant bacterial infections.

中文翻译：

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高通量微流体球状体技术，用于通过基于梯度的分析对粘菌素诱导的肾毒性进行早期检测。


粘菌素对于治疗多重耐药革兰氏阴性菌感染至关重要，但具有明显的肾毒性副作用。研究粘菌素肾毒性的传统方法受到其前药粘菌素甲磺酸盐的快速代谢以及难以从危重患者获得足够血浆的挑战。为了应对这些挑战，我们开发了球状肾毒性评估平台 （SNAP），这是一种微流控装置，可在 48 小时内仅使用 200 μL 患者血浆即可有效检测粘菌素诱导的肾近端肾小管上皮细胞 （RPTEC） 球状体中的毒性。我们的研究结果表明，与传统的二维 （2D） 培养物相比，SNAP 不仅促进肾脏特异性标志物水通道蛋白 1 （AQP1） 和低密度脂蛋白受体相关蛋白 2 （LRP2） 的更高表达，而且还表现出对粘菌素的敏感性增加，在 50 μg ml-1 及以上的浓度下检测到显着毒性。值得注意的是，SNAP 的非侵入性方法未识别健康供体血浆中的肾毒性，从而证实了其生理相关性，并显示出优于 2D 培养的敏感性，后者产生了假阳性结果。在临床验证中，SNAP 准确识别了有粘菌素诱导的肾毒性风险的患者，早发和晚发准确率均为 100%，预测肾毒性未发生率为 75%。这些结果强调了 SNAP 在个性化医疗中的潜力，为评估抗生素诱导的肾毒性提供了一种无创、精确和有效的工具，从而提高了针对耐药细菌感染的治疗的安全性和有效性。