Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators
The combination therapies ORKAMBI<sup>TM</sup> and TRIKAFTA<sup>TM</sup> are approved for people who have the F508del mutation on at least one allele. In this study we examine the effects of potentiator and corrector combinations on the rare mutation c.3700A>G. This mutati...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-11-01
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| Series: | Journal of Personalized Medicine |
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| Online Access: | https://www.mdpi.com/2075-4426/10/4/209 |
| _version_ | 1797548739903094784 |
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| author | Onofrio Laselva Jacqueline McCormack Claire Bartlett Wan Ip Tarini N. A. Gunawardena Hong Ouyang Paul D. W. Eckford Tanja Gonska Theo J. Moraes Christine E. Bear |
| author_facet | Onofrio Laselva Jacqueline McCormack Claire Bartlett Wan Ip Tarini N. A. Gunawardena Hong Ouyang Paul D. W. Eckford Tanja Gonska Theo J. Moraes Christine E. Bear |
| author_sort | Onofrio Laselva |
| collection | DOAJ |
| description | The combination therapies ORKAMBI<sup>TM</sup> and TRIKAFTA<sup>TM</sup> are approved for people who have the F508del mutation on at least one allele. In this study we examine the effects of potentiator and corrector combinations on the rare mutation c.3700A>G. This mutation produces a cryptic splice site that deletes six amino acids in NBD2 (I1234-R1239del). Like F508del it causes protein misprocessing and reduced chloride channel function. We show that a novel cystic fibrosis transmembrane conductance regulator CFTR modulator triple combination (AC1, corrector, AC2-2, co-potentiator and AP2, potentiator), rescued I1234-R1239del-CFTR activity to WT-CFTR level in HEK293 cells. Moreover, we show that although the response to ORKAMBI was modest in nasal epithelial cells from two individuals homozygous for I1234-R1239del-CFTR, a substantial functional rescue was achieved with the novel triple combination. Interestingly, while both the novel CFTR triple combination and TRIKAFTA<sup>TM</sup> treatment showed functional rescue in gene-edited I1234-R1239del-CFTR-expressing HBE cells and in nasal cells from two CF patients heterozygous for I1234-R1239del/W1282X, nasal cells homozygous for I1234-R1239del-CFTR showed no significant response to the TRIKAFTA<sup>TM</sup> combination. These data suggest a potential benefit of CFTR modulators on the functional rescue of I1234-R1239del -CFTR, which arises from the rare CF-causing mutation c.3700A>G, and highlight that patient tissues are crucial to our full understanding of functional rescue in rare CFTR mutations. |
| first_indexed | 2024-03-10T15:05:01Z |
| format | Article |
| id | doaj.art-52ee0e2f5fce41848629c9293ab25784 |
| institution | Directory Open Access Journal |
| issn | 2075-4426 |
| language | English |
| last_indexed | 2024-03-10T15:05:01Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Personalized Medicine |
| spelling | doaj.art-52ee0e2f5fce41848629c9293ab257842023-11-20T19:51:49ZengMDPI AGJournal of Personalized Medicine2075-44262020-11-0110420910.3390/jpm10040209Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR ModulatorsOnofrio Laselva0Jacqueline McCormack1Claire Bartlett2Wan Ip3Tarini N. A. Gunawardena4Hong Ouyang5Paul D. W. Eckford6Tanja Gonska7Theo J. Moraes8Christine E. Bear9Programme in Molecular Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Molecular Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Translational Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Translational Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Molecular Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Translational Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Molecular Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Translational Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Translational Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaProgramme in Molecular Medicine, Hospital for Sick Children, Toronto, ON M5G 8X4, CanadaThe combination therapies ORKAMBI<sup>TM</sup> and TRIKAFTA<sup>TM</sup> are approved for people who have the F508del mutation on at least one allele. In this study we examine the effects of potentiator and corrector combinations on the rare mutation c.3700A>G. This mutation produces a cryptic splice site that deletes six amino acids in NBD2 (I1234-R1239del). Like F508del it causes protein misprocessing and reduced chloride channel function. We show that a novel cystic fibrosis transmembrane conductance regulator CFTR modulator triple combination (AC1, corrector, AC2-2, co-potentiator and AP2, potentiator), rescued I1234-R1239del-CFTR activity to WT-CFTR level in HEK293 cells. Moreover, we show that although the response to ORKAMBI was modest in nasal epithelial cells from two individuals homozygous for I1234-R1239del-CFTR, a substantial functional rescue was achieved with the novel triple combination. Interestingly, while both the novel CFTR triple combination and TRIKAFTA<sup>TM</sup> treatment showed functional rescue in gene-edited I1234-R1239del-CFTR-expressing HBE cells and in nasal cells from two CF patients heterozygous for I1234-R1239del/W1282X, nasal cells homozygous for I1234-R1239del-CFTR showed no significant response to the TRIKAFTA<sup>TM</sup> combination. These data suggest a potential benefit of CFTR modulators on the functional rescue of I1234-R1239del -CFTR, which arises from the rare CF-causing mutation c.3700A>G, and highlight that patient tissues are crucial to our full understanding of functional rescue in rare CFTR mutations.https://www.mdpi.com/2075-4426/10/4/209cystic fibrosisCFTRrare mutationsc.3700A>GTRIKAFTA |
| spellingShingle | Onofrio Laselva Jacqueline McCormack Claire Bartlett Wan Ip Tarini N. A. Gunawardena Hong Ouyang Paul D. W. Eckford Tanja Gonska Theo J. Moraes Christine E. Bear Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators Journal of Personalized Medicine cystic fibrosis CFTR rare mutations c.3700A> G TRIKAFTA |
| title | Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators |
| title_full | Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators |
| title_fullStr | Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators |
| title_full_unstemmed | Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators |
| title_short | Preclinical Studies of a Rare CF-Causing Mutation in the Second Nucleotide Binding Domain (c.3700A>G) Show Robust Functional Rescue in Primary Nasal Cultures by Novel CFTR Modulators |
| title_sort | preclinical studies of a rare cf causing mutation in the second nucleotide binding domain c 3700a g show robust functional rescue in primary nasal cultures by novel cftr modulators |
| topic | cystic fibrosis CFTR rare mutations c.3700A> G TRIKAFTA |
| url | https://www.mdpi.com/2075-4426/10/4/209 |
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