Department of Molecular Biotechnology and Health Sciences, University of Turin, Via Nizza 52, Turin, 10125, Italy.
Department of Medical Sciences, University of Turin, Turin, Italy.
J Transl Med. 2024 Aug 14;22(1):762. doi: 10.1186/s12967-024-05575-z.
Personalized disease models are crucial for evaluating how diseased cells respond to treatments, especially in case of innovative biological therapeutics. Extracellular vesicles (EVs), nanosized vesicles released by cells for intercellular communication, have gained therapeutic interest due to their ability to reprogram target cells. We here utilized urinary podocytes obtained from children affected by steroid-resistant nephrotic syndrome with characterized genetic mutations as a model to test the therapeutic potential of EVs derived from kidney progenitor cells (nKPCs).
EVs were isolated from nKPCs derived from the urine of a preterm neonate. Three lines of urinary podocytes obtained from nephrotic patients' urine and a line of Alport syndrome patient podocytes were characterized and used to assess albumin permeability in response to nKPC-EVs or various drugs. RNA sequencing was conducted to identify commonly modulated pathways after nKPC-EV treatment. siRNA transfection was used to demonstrate the involvement of SUMO1 and SENP2 in the modulation of permeability.
Treatment with the nKPC-EVs significantly reduced permeability across all the steroid-resistant patients-derived and Alport syndrome-derived podocytes. At variance, podocytes appeared unresponsive to standard pharmacological treatments, with the exception of one line, in alignment with the patient's clinical response at 48 months. By RNA sequencing, only two genes were commonly upregulated in nKPC-EV-treated genetically altered podocytes: small ubiquitin-related modifier 1 (SUMO1) and Sentrin-specific protease 2 (SENP2). SUMO1 and SENP2 downregulation increased podocyte permeability confirming the role of the SUMOylation pathway.
nKPCs emerge as a promising non-invasive source of EVs with potential therapeutic effects on podocytes with genetic dysfunction, through modulation of SUMOylation, an important pathway for the stability of podocyte slit diaphragm proteins. Our findings also suggest the feasibility of developing a non-invasive in vitro model for screening regenerative compounds on patient-derived podocytes.
个性化疾病模型对于评估患病细胞对治疗的反应至关重要，特别是在创新的生物治疗药物的情况下。细胞间通讯释放的纳米大小的囊泡细胞外囊泡 (EVs)，由于能够重新编程靶细胞，因此具有治疗潜力。我们利用从患有类固醇耐药性肾病综合征的儿童中获得的尿足细胞作为模型，来测试源自肾祖细胞 (nKPC) 的 EV 的治疗潜力。
从早产儿尿液中分离出 nKPC 衍生的 EV。对从肾病患者尿液中获得的 3 条尿足细胞系和一条 Alport 综合征患者足细胞系进行了特征描述，并用于评估对 nKPC-EV 或各种药物的白蛋白通透性。进行 RNA 测序以鉴定 nKPC-EV 处理后共同调节的途径。使用 siRNA 转染来证明 SUMO1 和 SENP2 在通透性调节中的作用。
用 nKPC-EV 处理可显著降低所有类固醇耐药患者来源和 Alport 综合征来源的足细胞的通透性。相比之下，除了一条与患者在 48 个月时的临床反应一致的线外，足细胞对标准药物治疗似乎没有反应。通过 RNA 测序，只有两个基因在 nKPC-EV 处理的遗传改变的足细胞中共同上调：小泛素相关修饰物 1 (SUMO1) 和 Sentrin 特异性蛋白酶 2 (SENP2)。SUMO1 和 SENP2 的下调增加了足细胞的通透性，证实了 SUMOylation 途径的作用。
nKPC 作为 EV 的一种有前途的非侵入性来源，通过调节 SUMOylation，对具有遗传功能障碍的足细胞具有潜在的治疗效果，SUMOylation 是足细胞裂孔隔膜蛋白稳定性的重要途径。我们的研究结果还表明，开发用于在患者来源的足细胞上筛选再生化合物的非侵入性体外模型是可行的。
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