Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice
Cantu syndrome (CS) is caused by gain-of-function (GOF) mutations in pore-forming (Kir6.1, KCNJ8) and accessory (SUR2, ABCC9) ATP-sensitive potassium (KATP) channel subunits, the most common mutations being SUR2[R1154Q] and SUR2[R1154W], carried by approximately 30% of patients. We used CRISPR/Cas9...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Clinical investigation
2021-03-01
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| Series: | JCI Insight |
| Subjects: | |
| Online Access: | https://doi.org/10.1172/jci.insight.145934 |
| _version_ | 1818858866779095040 |
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| author | Haixia Zhang Alex Hanson Tobias Scherf de Almeida Christopher Emfinger Conor McClenaghan Theresa Harter Zihan Yan Paige E. Cooper G. Schuyler Brown Eric C. Arakel Robert P. Mecham Atilla Kovacs Carmen M. Halabi Blanche Schwappach Maria S. Remedi Colin G. Nichols |
| author_facet | Haixia Zhang Alex Hanson Tobias Scherf de Almeida Christopher Emfinger Conor McClenaghan Theresa Harter Zihan Yan Paige E. Cooper G. Schuyler Brown Eric C. Arakel Robert P. Mecham Atilla Kovacs Carmen M. Halabi Blanche Schwappach Maria S. Remedi Colin G. Nichols |
| author_sort | Haixia Zhang |
| collection | DOAJ |
| description | Cantu syndrome (CS) is caused by gain-of-function (GOF) mutations in pore-forming (Kir6.1, KCNJ8) and accessory (SUR2, ABCC9) ATP-sensitive potassium (KATP) channel subunits, the most common mutations being SUR2[R1154Q] and SUR2[R1154W], carried by approximately 30% of patients. We used CRISPR/Cas9 genome engineering to introduce the equivalent of the human SUR2[R1154Q] mutation into the mouse ABCC9 gene. Along with minimal CS disease features, R1154Q cardiomyocytes and vascular smooth muscle showed much lower KATP current density and pinacidil activation than WT cells. Almost complete loss of SUR2-dependent protein and KATP in homozygous R1154Q ventricles revealed underlying diazoxide-sensitive SUR1-dependent KATP channel activity. Surprisingly, sequencing of SUR2 cDNA revealed 2 distinct transcripts, one encoding full-length SUR2 protein; and the other with an in-frame deletion of 93 bases (corresponding to 31 amino acids encoded by exon 28) that was present in approximately 40% and approximately 90% of transcripts from hetero- and homozygous R1154Q tissues, respectively. Recombinant expression of SUR2A protein lacking exon 28 resulted in nonfunctional channels. CS tissue from SUR2[R1154Q] mice and human induced pluripotent stem cell–derived (hiPSC-derived) cardiomyocytes showed only full-length SUR2 transcripts, although further studies will be required in order to fully test whether SUR2[R1154Q] or other CS mutations might result in aberrant splicing and variable expressivity of disease features in human CS. |
| first_indexed | 2024-12-19T09:03:06Z |
| format | Article |
| id | doaj.art-49205ba65db14c0c92f39428e2cf79af |
| institution | Directory Open Access Journal |
| issn | 2379-3708 |
| language | English |
| last_indexed | 2024-12-19T09:03:06Z |
| publishDate | 2021-03-01 |
| publisher | American Society for Clinical investigation |
| record_format | Article |
| series | JCI Insight |
| spelling | doaj.art-49205ba65db14c0c92f39428e2cf79af2022-12-21T20:28:25ZengAmerican Society for Clinical investigationJCI Insight2379-37082021-03-0165Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified miceHaixia ZhangAlex HansonTobias Scherf de AlmeidaChristopher EmfingerConor McClenaghanTheresa HarterZihan YanPaige E. CooperG. Schuyler BrownEric C. ArakelRobert P. MechamAtilla KovacsCarmen M. HalabiBlanche SchwappachMaria S. RemediColin G. NicholsCantu syndrome (CS) is caused by gain-of-function (GOF) mutations in pore-forming (Kir6.1, KCNJ8) and accessory (SUR2, ABCC9) ATP-sensitive potassium (KATP) channel subunits, the most common mutations being SUR2[R1154Q] and SUR2[R1154W], carried by approximately 30% of patients. We used CRISPR/Cas9 genome engineering to introduce the equivalent of the human SUR2[R1154Q] mutation into the mouse ABCC9 gene. Along with minimal CS disease features, R1154Q cardiomyocytes and vascular smooth muscle showed much lower KATP current density and pinacidil activation than WT cells. Almost complete loss of SUR2-dependent protein and KATP in homozygous R1154Q ventricles revealed underlying diazoxide-sensitive SUR1-dependent KATP channel activity. Surprisingly, sequencing of SUR2 cDNA revealed 2 distinct transcripts, one encoding full-length SUR2 protein; and the other with an in-frame deletion of 93 bases (corresponding to 31 amino acids encoded by exon 28) that was present in approximately 40% and approximately 90% of transcripts from hetero- and homozygous R1154Q tissues, respectively. Recombinant expression of SUR2A protein lacking exon 28 resulted in nonfunctional channels. CS tissue from SUR2[R1154Q] mice and human induced pluripotent stem cell–derived (hiPSC-derived) cardiomyocytes showed only full-length SUR2 transcripts, although further studies will be required in order to fully test whether SUR2[R1154Q] or other CS mutations might result in aberrant splicing and variable expressivity of disease features in human CS.https://doi.org/10.1172/jci.insight.145934Muscle biologyVascular biology |
| spellingShingle | Haixia Zhang Alex Hanson Tobias Scherf de Almeida Christopher Emfinger Conor McClenaghan Theresa Harter Zihan Yan Paige E. Cooper G. Schuyler Brown Eric C. Arakel Robert P. Mecham Atilla Kovacs Carmen M. Halabi Blanche Schwappach Maria S. Remedi Colin G. Nichols Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice JCI Insight Muscle biology Vascular biology |
| title | Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice |
| title_full | Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice |
| title_fullStr | Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice |
| title_full_unstemmed | Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice |
| title_short | Complex consequences of Cantu syndrome SUR2 variant R1154Q in genetically modified mice |
| title_sort | complex consequences of cantu syndrome sur2 variant r1154q in genetically modified mice |
| topic | Muscle biology Vascular biology |
| url | https://doi.org/10.1172/jci.insight.145934 |
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