Characterization of the mechanism of action of RDR01752, a novel corrector of F508del-CFTR

Lopes‐Pacheco, Miquéias; Silva, Iris; Turner, Mark J.; Carlile, Graeme W.; Sondo, Elvira; Thomas, David Y.; Pedemonte, Nicoletta; Hanrahan, John W.; Amaral, Margarida D.

doi:10.1016/j.bcp.2020.114133

Cited by 14 publications

(19 citation statements)

References 38 publications

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“…CF bronchial epithelial (CFBE) cell lines constitutively expressing CFTR variants (WT, DD/AA, F508del, R560S, or carrying G550E, R1070W, or 4RK in cis with F508del) were cultured in Eagle’s minimum essential medium (EMEM, #10-010-CVR, Corning, VA, USA) supplemented with 10% FBS and 2 µg/mL puromycin [ 14 , 24 ]. CFBE cells co-expressing the halide sensitive yellow fluorescence protein (HS-YFP H148Q/I152L) with either WT- or F508del-CFTR were cultured in EMEM supplemented with 10% FBS, and selection antibiotics (0.5 µg/mL puromycin for WT-CFTR-expressing cells and 2 µg/mL puromycin plus 0.6 mg/mL G418 for F508del-expressing ones) [ 31 , 32 ].…”

Section: Methodsmentioning

confidence: 99%

“…This mutation impairs NBD1 thermodynamic stability and its interdomain interactions, causing CFTR protein misfolding that is disqualified from exiting the endoplasmic reticulum (ER) by the quality control (ERQC), being, thus, precluded from PM trafficking and targeted for proteasomal degradation [ 5 , 6 ]. Although F508del-CFTR folding is inefficient, it can be, nevertheless, rescued when cells heterologously expressing this mutant are incubated at a low temperature [ 7 , 8 , 9 , 10 ] or by CFTR genetic revertants, i.e., second-site mutations in cis with F508del that partially rescue this mutant either by correcting folding by filling in its structural pockets (absent in WT-CFTR), or by removing retention signals, thus allowing the protein to evade ERQC [ 11 , 12 , 13 , 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

“…The MoA of MCG1516A was investigated by examining its additive effects to low temperature incubation and to previously established CFTR genetic revertants. These included G550E that restores NBD1:NBD2 dimerization [ 12 , 14 ], R1070W that retrieves NBD1:ICL4 interface [ 13 , 15 ], and 4RK (R29K, R516K, R555K, and R766K) that simultaneously withdraws four arginine-framed tripeptides (AFT) retention signals [ 11 , 13 ]. We also assessed the effects of MCG1516A on a CFTR variant lacking the double diacidic code (DD/AA), which is, thus, precluded from exiting the ER via COPII vesicles, although it does not present a major conformational defect [ 13 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

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Rescue of Mutant CFTR Trafficking Defect by the Investigational Compound MCG1516A

Lopes‐Pacheco

Bacalhau

Ramalho

et al. 2022

Cells

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Although some therapeutic progress has been achieved in developing small molecules that correct F508del-CFTR defects, the mechanism of action (MoA) of these compounds remain poorly elucidated. Here, we investigated the effects and MoA of MCG1516A, a newly developed F508del-CFTR corrector. MCG1516A effects on wild-type (WT) and F508del-CFTR were assessed by immunofluorescence microscopy, and biochemical and functional assays both in cell lines and in intestinal organoids. To shed light on the MoA of MCG1516A, we evaluated its additivity to the FDA-approved corrector VX-661, low temperature, genetic revertants of F508del-CFTR (G550E, R1070W, and 4RK), and the traffic-null variant DD/AA. Finally, we explored the ability of MCG1516A to rescue trafficking and function of other CF-causing mutations. We found that MCG1516A rescues F508del-CFTR with additive effects to VX-661. A similar behavior was observed for WT-CFTR. Under low temperature incubation, F508del-CFTR demonstrated an additivity in processing and function with VX-661, but not with MCG1516A. In contrast, both compounds promoted additional effects to low temperature to WT-CFTR. MCG1516A demonstrated additivity to genetic revertant R1070W, while VX-661 was additive to G550E and 4RK. Nevertheless, none of these compounds rescued DD/AA trafficking. Both MCG1516A and VX-661 rescued CFTR processing of L206W- and R334W-CFTR with greater effects when these compounds were combined. In summary, the absence of additivity of MCG1516A to genetic revertant G550E suggests a putative binding site for this compound on NBD1:NBD2 interface. Therefore, a combination of MCG1516A with compounds able to rescue DD/AA traffic, or mimicking the actions of revertant R1070W (e.g., VX-661), could enhance correction of F508del-CFTR defects.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Rescue of Mutant CFTR Trafficking Defect by the Investigational Compound MCG1516A

Lopes‐Pacheco

Bacalhau

Ramalho

et al. 2022

Cells

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“…Other investigational correctors include FDL-169 (Flatley Discovery) 68 and RDR01752, 78 which also appear to share the rescue mechanism with VX-809 and VX-661. In addition, three small-molecule series were identified in a HTS of ~600,000 drug-like molecules: 6258, 3151 and 4172, which target defects at NBD1, NBD2 and TMD interfaces, respectively.…”

Section: Cftr Modulator Drugs and Personalized Medicinementioning

confidence: 99%

“…76 Abbvie also has two additional investigational correctors (AC1 and AC2) that were shown to rescue processing and trafficking of other class II CFTR mutations, including G85E, M1101K and N1303K. 46,77 Other investigational correctors include FDL-169 (Flatley Discovery) 68 and RDR01752, 78 which also appear to share the rescue mechanism with VX-809 and VX-661. In addition, three small-molecule series were identified in a HTS of ~600,000 drug-like molecules: 6258, 3151 and 4172, which target defects at NBD1, NBD2 and TMD interfaces, respectively.…”

Section: Class II Mutations: Correctorsmentioning

confidence: 99%

Pharmacological Modulation of Ion Channels for the Treatment of Cystic Fibrosis

Pinto¹,

Silva²,

Figueira³

et al. 2021

JEP

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Cystic fibrosis (CF) is a life-shortening monogenic disease caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR) protein, an anion channel that transports chloride and bicarbonate across epithelia. Despite clinical progress in delaying disease progression with symptomatic therapies, these individuals still develop various chronic complications in lungs and other organs, which significantly restricts their life expectancy and quality of life. The development of high-throughput assays to screen drug-like compound libraries have enabled the discovery of highly effective CFTR modulator therapies. These novel therapies target the primary defect underlying CF and are now approved for clinical use for individuals with specific CF genotypes. However, the clinically approved modulators only partially reverse CFTR dysfunction and there is still a considerable number of individuals with CF carrying rare CFTR mutations who remain without any effective CFTR modulator therapy. Accordingly, additional efforts have been pursued to identify novel and more potent CFTR modulators that may benefit a larger CF population. The use of ex vivo individual-derived specimens has also become a powerful tool to evaluate novel drugs and predict their effectiveness in a personalized medicine approach. In addition to CFTR modulators, pro-drugs aiming at modulating alternative ion channels/transporters are under development to compensate for the lack of CFTR function. These therapies may restore normal mucociliary clearance through a mutation-agnostic approach (ie, independent of CFTR mutation) and include inhibitors of the epithelial sodium channel (ENaC), modulators of the calcium-activated channel transmembrane 16A (TMEM16, or anoctamin 1) or of the solute carrier family 26A member 9 (SLC26A9), and anionophores. The present review focuses on recent progress and challenges for the development of ion channel/transporter-modulating drugs for the treatment of CF.

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The use of pharmacological chaperones in rare diseases caused by reduced protein stability

et al. 2022

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Rare diseases are most often caused by inherited genetic disorders that, after translation, will result in a protein with altered function. Decreased protein stability is the most frequent mechanism associated with a congenital pathogenic missense mutation and it implies the destabilization of the folded conformation in favour of unfolded or misfolded states. In the cellular context and when experimental data is available, a mutant protein with altered thermodynamic stability often also results in impaired homeostasis, with the deleterious accumulation of protein aggregates, metabolites and/or metabolic by‐products. In the last decades, a significant effort has enabled the characterization of rare diseases associated to protein stability defects and triggered the development of innovative therapeutic intervention lines, say, the use of pharmacological chaperones to correct the intracellular impaired homeostasis. Here, we review the current knowledge on rare diseases caused by reduced protein stability, paying special attention to the thermodynamic aspects of the protein destabilization, also focusing on some examples where pharmacological chaperones are being tested.

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Characterization of the mechanism of action of RDR01752, a novel corrector of F508del-CFTR

Cited by 14 publications

References 38 publications

Rescue of Mutant CFTR Trafficking Defect by the Investigational Compound MCG1516A

Rescue of Mutant CFTR Trafficking Defect by the Investigational Compound MCG1516A

Pharmacological Modulation of Ion Channels for the Treatment of Cystic Fibrosis

The use of pharmacological chaperones in rare diseases caused by reduced protein stability

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