Abstract：
Nephropathic cystinosis is a rare and severe disease caused by disruptions in the CTNS gene. Cystinosis is characterized by lysosomal cystine accumulation, vesicle trafficking impairment, oxidative stress, and apoptosis. Additionally, cystinotic patients exhibit weakening and leakage of the proximal tubular segment of the nephrons, leading to renal Fanconi syndrome and kidney failure early in life. Current in vitro cystinotic models cannot recapitulate all clinical features of the disease which limits their translational value. Therefore, the development of novel, complex in vitro models that better mimic the disease and exhibit characteristics not compatible with 2-dimensional cell culture is of crucial importance for novel therapies development. In this study, we developed a 3-dimensional bioengineered model of nephropathic cystinosis by culturing conditionally immortalized proximal tubule epithelial cells (ciPTECs) on hollow fiber membranes (HFM). Cystinotic kidney tubules showed lysosomal cystine accumulation, increased autophagy and vesicle trafficking deterioration, the impairment of several metabolic pathways, and the disruption of the epithelial monolayer tightness as compared to control kidney tubules. In particular, the loss of monolayer organization and leakage could be mimicked with the use of the cystinotic kidney tubules, which has not been possible before, using the standard 2-dimensional cell culture. Overall, bioengineered cystinotic kidney tubules recapitulate better the nephropathic phenotype at a molecular, structural, and functional proximal tubule level compared to 2-dimensional cell cultures.
摘要：
肾病性胱氨酸病是一种由CTNS基因破坏引起的罕见且严重的疾病。胱抑素病以溶酶体胱氨酸积累为特征，囊泡运输损害，氧化应激，和凋亡。此外,囊肿患者表现出肾单位近端管状节段的减弱和渗漏，在生命早期导致肾性范可尼综合征和肾衰竭。目前的体外膀胱囊肿模型无法概括该疾病的所有临床特征，这限制了其翻译价值。因此,小说的发展,复杂的体外模型可以更好地模拟疾病并表现出与二维细胞培养不相容的特征，这对于新疗法的开发至关重要。在这项研究中,我们通过在中空纤维膜（HFM）上培养有条件永生化的近端小管上皮细胞（ciPTEC），建立了肾病性膀胱炎的3维生物工程模型。囊肿性肾小管显示溶酶体胱氨酸积聚，自噬增加和囊泡运输恶化，几种代谢途径的损害，与对照肾小管相比，上皮单层紧密度的破坏。特别是,单层组织的丧失和渗漏可以通过使用囊肿性肾小管来模仿，这在以前是不可能的，使用标准的二维细胞培养。总的来说,生物工程制囊性肾小管在分子上更好地概括了肾病表型，结构,与二维细胞培养物相比，功能近端小管水平。
