NBD2 Is Required for the Rescue of Mutant F508del CFTR by a Thiazole-Based Molecule: A Class II Corrector for the Multi-Drug Therapy of Cystic Fibrosis

Brandas, Chiara; Ludovico, Alessandra; Parodi, Alice; Morán, Óscar; Millo, Enrico; Cichero, Elena; Baroni, Debora

doi:10.3390/biom11101417

Cited by 9 publications

(24 citation statements)

References 66 publications

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“…Another aspect of the interaction between VX445 and F508del that we sought to address in this work was whether VX445 had an effect on the stability of MSD2. Corroborated by previous results obtained with other CFTR modulators [34,45], we hypothesized that if VX445 acts by stabilizing the region whose expression it increases, then it could be expected that this region would have a slower turnover rate after transiently transfected cells are treated with cycloheximide to inhibit protein synthesis. Therefore, HEK-t cells were transfected with plasmids encoding MSD2 and incubated in the presence of VX445, VX809, VX661, CORR4A, or DMSO.…”

Section: Discussionmentioning

confidence: 83%

“…In this work, we applied different biochemical approaches to identify the CFTR domain mainly involved in the rescue of mutant F508del protein expression promoted by VX445. To achieve this aim, we heterologously expressed plasmid constructs encoding the full-length F508del and MSD1, WT and F508del NBD1, the R domain, and the NBD2 and MSD2 isolated domains into highly transfectable human embryonic kidney 293 (HEK-t) cells and analyzed the effect of VX445 on their expression and stability, comparing it with that exerted by other CFTR correctors whose molecular mechanism of action has been previously described [ 32 , 33 , 34 , 43 , 44 , 45 ]. Our analysis identified MSD2 as the CFTR domain mainly involved in the rescue of F508del CFTR promoted by VX445.…”

Section: Introductionmentioning

confidence: 99%

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Elexacaftor Mediates the Rescue of F508del CFTR Functional Expression Interacting with MSD2

Bongiorno

Ludovico

Morán

et al. 2023

IJMS

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Cystic fibrosis (CF) is one of the most frequent lethal autosomal recessive diseases affecting the Caucasian population. It is caused by loss of function variants of the cystic fibrosis transmembrane conductance regulator (CFTR), a membrane protein located on the apical side of epithelial cells. The most prevalent CF-causing mutation, the deletion of phenylalanine at position 508 (F508del), is characterized by folding and trafficking defects, resulting in the decreased functional expression of the protein on the plasma membrane. Two classes of small-molecule modulators, termed potentiators and correctors, respectively, have been developed to rescue either the gating or the cellular processing of defective F508del CFTR. Kaftrio, a next-generation triple-combination drug, consisting of the potentiator ivacaftor (VX770) and the two correctors tezacaftor (VX661) and elexacaftor (VX445), has been demonstrated to be a life-changing therapeutic modality for the majority of people with CF worldwide. While the mechanism of action of VX770 and VX661 is almost known, the precise mechanism of action and binding site of VX445 have not been conclusively determined. We investigated the activity of VX445 on mutant F508del to identify the protein domains whose expression is mostly affected by this corrector and to disclose its mechanisms of action. Our biochemical analyses revealed that VX445 specifically improves the expression and the maturation of MSD2, heterologously expressed in HEK 293 cells, and confirmed that its effect on the functional expression of defective F508del CFTR is additive either with type I or type II CFTR correctors. We are confident that our study will help to make a step forward in the comprehension of the etiopathology of the CF disease, as well as to give new information for the development and testing of combinations of even more effective correctors able to target mutation-specific defects of the CFTR protein.

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Section: Discussionmentioning

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Elexacaftor Mediates the Rescue of F508del CFTR Functional Expression Interacting with MSD2

Bongiorno

Ludovico

Morán

et al. 2023

IJMS

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“…Corr-4a remains the archetypical example of type II correctors (i.e., targeting NBD2-MSD2) [ 123 ]. Other correctors of this type, such as the aminoarylthiazole derivative, FCG, were recently proposed to bind to NBD2 [ 136 , 137 ]. The MoA of type II correctors is complementary to that of other corrector types and hence allows to be added in conjunction ( Figure 5 ) [ 120 , 123 , 138 , 139 ].…”

Section: Cftr Causal Therapiesmentioning

confidence: 99%

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Ensinck

Carlon

2022

Cells

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Cystic fibrosis (CF) is the most common monogenic disorder, caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Over the last 30 years, tremendous progress has been made in understanding the molecular basis of CF and the development of treatments that target the underlying defects in CF. Currently, a highly effective CFTR modulator treatment (Kalydeco™/Trikafta™) is available for 90% of people with CF. In this review, we will give an extensive overview of past and ongoing efforts in the development of therapies targeting the molecular defects in CF. We will discuss strategies targeting the CFTR protein (i.e., CFTR modulators such as correctors and potentiators), its cellular environment (i.e., proteostasis modulation, stabilization at the plasma membrane), the CFTR mRNA (i.e., amplifiers, nonsense mediated mRNA decay suppressors, translational readthrough inducing drugs) or the CFTR gene (gene therapies). Finally, we will focus on how these efforts can be applied to the 15% of people with CF for whom no causal therapy is available yet.

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“…Since the approval of TEZ/IVA for S945L, the triple combination modulator therapy ELX/TEZ/IVA has expanded treatment access to more than 90% of people with CF and is currently the most effective treatment for improving patient lung function ( 19 , 64 ). Additionally, the potential utility of novel co-potentiators ( 65 , 66 ), correctors ( 67 – 69 ), amplifiers ( 70 ), and a drug targeting the misfolding detection machinery ( 71 ) have been assessed. Since the participant enrolled in this study was receiving treatment with TEZ/IVA, the focus was to compare the participant's in vitro response to TEZ/IVA with their in vivo response to TEZ/IVA .…”

Section: Discussionmentioning

confidence: 99%

S945L-CFTR molecular dynamics, functional characterization and tezacaftor/ivacaftor efficacy in vivo and in vitro in matched pediatric patient-derived cell models

et al. 2022

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Cystic Fibrosis (CF) results from over 400 different disease-causing mutations in the CF Transmembrane Conductance Regulator (CFTR) gene. These CFTR mutations lead to numerous defects in CFTR protein function. A novel class of targeted therapies (CFTR modulators) have been developed that can restore defects in CFTR folding and gating. This study aimed to characterize the functional and structural defects of S945L-CFTR and interrogate the efficacy of modulators with two modes of action: gating potentiator [ivacaftor (IVA)] and folding corrector [tezacaftor (TEZ)]. The response to these modulators in vitro in airway differentiated cell models created from a participant with S945L/G542X-CFTR was correlated with in vivo clinical outcomes of that participant at least 12 months pre and post modulator therapy. In this participants' airway cell models, CFTR-mediated chloride transport was assessed via ion transport electrophysiology. Monotherapy with IVA or TEZ increased CFTR activity, albeit not reaching statistical significance. Combination therapy with TEZ/IVA significantly (p = 0.02) increased CFTR activity 1.62-fold above baseline. Assessment of CFTR expression and maturation via western blot validated the presence of mature, fully glycosylated CFTR, which increased 4.1-fold in TEZ/IVA-treated cells. The in vitro S945L-CFTR response to modulator correlated with an improvement in in vivo lung function (ppFEV1) from 77.19 in the 12 months pre TEZ/IVA to 80.79 in the 12 months post TEZ/IVA. The slope of decline in ppFEV1 significantly (p = 0.02) changed in the 24 months post TEZ/IVA, becoming positive. Furthermore, there was a significant improvement in clinical parameters and a fall in sweat chloride from 68 to 28 mmol/L. The mechanism of dysfunction of S945L-CFTR was elucidated by in silico molecular dynamics (MD) simulations. S945L-CFTR caused misfolding of transmembrane helix 8 and disruption of the R domain, a CFTR domain critical to channel gating. This study showed in vitro and in silico that S945L causes both folding and gating defects in CFTR and demonstrated in vitro and in vivo that TEZ/IVA is an efficacious modulator combination to address these defects. As such, we support the utility of patient-derived cell models and MD simulations in predicting and understanding the effect of modulators on CFTR function on an individualized basis.

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NBD2 Is Required for the Rescue of Mutant F508del CFTR by a Thiazole-Based Molecule: A Class II Corrector for the Multi-Drug Therapy of Cystic Fibrosis

Cited by 9 publications

References 66 publications

Elexacaftor Mediates the Rescue of F508del CFTR Functional Expression Interacting with MSD2

Elexacaftor Mediates the Rescue of F508del CFTR Functional Expression Interacting with MSD2

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

S945L-CFTR molecular dynamics, functional characterization and tezacaftor/ivacaftor efficacy in vivo and in vitro in matched pediatric patient-derived cell models

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