Targeting autophagy as a novel strategy for facilitating the therapeutic action of potentiators on ΔF508 cystic fibrosis transmembrane conductance regulator
Targeting autophagy as a novel strategy for facilitating the therapeutic action of potentiators on ΔF508 cystic fibrosis transmembrane conductance regulator
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DOI：
10.4161/auto.21483
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发表时间：
2012-11-01
期刊：
影响因子：
13.3
通讯作者：
Maiuri, Luigi
中科院分区：
文献类型：
--
作者：
Luciani, Alessandro;Villella, Valeria Rachela;Maiuri, Luigi
囊性纤维化（CF）跨膜传导调节因子（CFTR）的通道激活剂（增强剂）可用于治疗携带质膜驻留CFTR突变体的CF患者的小子集。然而，大约90%的CF患者携带错误折叠的Delta F508-CFTR，并且对增效剂反应不良，因为Delta F508-CFTR在质膜（PM）处本质上是不稳定的，即使被药理学校正剂挽救。我们已经证明，人类和小鼠CF气道由于BECN 1的功能性隔离而缺乏自噬，并且组织转氨酶-2抑制剂、胱胺或抗氧化剂恢复BECN 1依赖性自噬并降低SQSTM 1/p62水平，从而有利于Delta F508-CFTR运输至上皮表面。在这里，我们研究了这些治疗是否可以促进增效剂对Delta F508-CFTR纯合气道的有益作用。胱胺或超氧化物歧化酶（SOD）/过氧化氢酶模拟物EUK-134稳定气道上皮细胞质膜处的Delta F508-CFTR，并维持上皮表面处CFTR的表达远远超过药物戒断，过表达BECN 1并消耗SQSTM 1。这促进了增效剂在控制离体Delta F508-CFTR纯合人鼻活组织检查和体内小鼠Delta F508-CFTR肺中的炎症中的有益作用。在Delta F508-CFTR小鼠体内通过shRNA直接消耗Sqstm 1与增效剂协同维持表面CFTR表达并抑制炎症。在来自Delta F508-CFTR纯合患者的新鲜分离的刷过的鼻上皮细胞中，胱胺预处理恢复了Delta F508-CFTR对CFTR增强剂染料木素、VEGF-532或VEGF-770的应答。这些发现描绘了一种新的治疗策略，用于治疗具有Delta F508-CFTR突变的CF患者，其中患者首先用胱胺治疗，随后用CFTR增效剂脉冲。
Channel activators (potentiators) of cystic fibrosis (CF) transmembrane conductance regulator (CFTR), can be used for the treatment of the small subset of CF patients that carry plasma membrane-resident CFTR mutants. However, approximately 90% of CF patients carry the misfolded Delta F508-CFTR and are poorly responsive to potentiators, because Delta F508-CFTR is intrinsically unstable at the plasma membrane (PM) even if rescued by pharmacological correctors. We have demonstrated that human and mouse CF airways are autophagy deficient due to functional sequestration of BECN1 and that the tissue transglutaminase-2 inhibitor, cystamine, or antioxidants restore BECN1-dependent autophagy and reduce SQSTM1/p62 levels, thus favoring Delta F508-CFTR trafficking to the epithelial surface. Here, we investigated whether these treatments could facilitate the beneficial action of potentiators on Delta F508-CFTR homozygous airways. Cystamine or the superoxide dismutase (SO D)/catalase-mimetic EUK-134 stabilized Delta F508-CFTR at the plasma membrane of airway epithelial cells and sustained the expression of CFTR at the epithelial surface well beyond drug withdrawal, overexpressing BECN1 and depleting SQSTM1. This facilitates the beneficial action of potentiators in controlling inflammation in ex vivo Delta F508-CFTR homozygous human nasal biopsies and in vivo in mouse Delta F508-CFTR lungs. Direct depletion of Sqstm1 by shRNAs in vivo in Delta F508-CFTR mice synergized with potentiators in sustaining surface CFTR expression and suppressing inflammation. Cystamine pre-treatment restored Delta F508-CFTR response to the CFTR potentiators genistein, Vrx-532 or Vrx-770 in freshly isolated brushed nasal epithelial cells from Delta F508-CFTR homozygous patients. These findings delineate a novel therapeutic strategy for the treatment of CF patients with the Delta F508-CFTR mutation in which patients are first treated with cystamine and subsequently pulsed with CFTR potentiators.