背景:我们的目标是使用离体人体微生理系统(MPS)发现抗生素相关肾毒性的新型尿生物标志物,并将这些发现转化为接受氨基糖苷类和/或多粘菌素E(粘菌素)治疗肺加重的前瞻性纳入囊性纤维化(CF)人群。
方法:我们用来自三个供体的原发性人肾近端小管上皮细胞(PTEC)填充MPS,并通过暴露于50µg/mL多粘菌素E72小时来模拟肾毒素损伤。我们通过RNAseq分析了基因转录反应并测试了MPS流出物。我们通过分析之前收集的尿液将候选生物标志物翻译为CF队列,在使用抗生素期间和使用抗生素后2周,对患者进行中位随访3年.
结果:多粘菌素E处理导致促凋亡Fas基因相对于对照在MPS的RNAseq中的统计学显著增加:倍数变化=1.63,FDRq值=7.29×10-5。流出物分析显示与细胞损伤相关的可溶性Fas(sFas)浓度的急性升高。在16例CF患者中,在抗生素治疗期间,尿sFas浓度显着升高,不管AKI的发展。经过三年的平均随访,我们确定了7例慢性肾脏病(CKD)偶发病例.抗生素治疗期间的尿sFas浓度与随后发生CKD显著相关(未调整的相对风险=2.02每增加一次尿sFas,95%CI=1.40,2.90,p<0.001)。
结论:使用离体MPS,我们发现了一种新的近端小管上皮细胞损伤的生物标志物,sFas,并将这些发现转化为CF患者的临床队列。
BACKGROUND: Our objective was to discover novel urinary biomarkers of antibiotic-associated nephrotoxicity using an ex-vivo human microphysiological system (MPS) and to translate these findings to a prospectively enrolled cystic fibrosis (CF) population receiving aminoglycosides and/or polymyxin E (colistin) for a pulmonary exacerbation.
METHODS: We populated the MPS with primary human kidney proximal tubule epithelial cells (PTECs) from three donors and modeled nephrotoxin injury through exposure to 50 µg/mL polymyxin E for 72 h. We analyzed gene transcriptional responses by RNAseq and tested MPS effluents. We translated candidate biomarkers to a CF cohort via analysis of urine collected prior to, during and two weeks after antibiotics and patients were followed for a median of 3 years after antibiotic use.
RESULTS: Polymyxin E treatment resulted in a statistically significant increase in the pro-apoptotic Fas gene relative to control in RNAseq of MPS: fold-change = 1.63, FDR q-value = 7.29 × 10-5. Effluent analysis demonstrated an acute rise of soluble Fas (sFas) concentrations that correlated with cellular injury. In 16 patients with CF, urinary sFas concentrations were significantly elevated during antibiotic treatment, regardless of development of AKI. Over a median of three years of follow up, we identified seven cases of incident chronic kidney disease (CKD). Urinary sFas concentrations during antibiotic treatment were significantly associated with subsequent development of incident CKD (unadjusted relative risk = 2.02 per doubling of urinary sFas, 95 % CI = 1.40, 2.90, p < 0.001).
CONCLUSIONS: Using an ex-vivo MPS, we identified a novel biomarker of proximal tubule epithelial cell injury, sFas, and translated these findings to a clinical cohort of patients with CF.