Abstract
Genetic variants that cause haploinsufficiency account for many autosomal dominant (AD) disorders. Gene-based diagnosis classifies variants that alter canonical splice signals as pathogenic, but due to imperfect understanding of RNA splice signals other variants that may create or eliminate splice sites are often clinically classified as variants of unknown significance (VUS). To improve recognition of pathogenic splice-altering variants in AD disorders, we used computational tools to prioritize VUS and developed a cell-based minigene splicing assay to confirm aberrant splicing. Using this two-step procedure we evaluated all rare variants in two AD cardiomyopathy genes, lamin A/C (LMNA) and myosin binding protein C (MYBPC3). We demonstrate that 13 LMNA and 35 MYBPC3 variants identified in cardiomyopathy patients alter RNA splicing, representing a 50% increase in the numbers of established damaging splice variants in these genes. Over half of these variants are annotated as VUS by clinical diagnostic laboratories. Familial analyses of one variant, a synonymous LMNA VUS, demonstrated segregation with cardiomyopathy affection status and altered cardiac LMNA splicing. Application of this strategy should improve diagnostic accuracy and variant classification in other haploinsufficient AD disorders.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 7689-7694 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 114 |
| Issue number | 29 |
| DOIs | |
| State | Published - Jul 18 2017 |
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All Science Journal Classification (ASJC) codes
- General
Cite this
Identification of pathogenic gene mutations in LMNA and MYBPC3 that alter RNA splicing. / Ito, Kaoru; Patel, Parth N.; Gorham, Joshua M.; McDonough, Barbara; DePalma, Steven R.; Adler, Emily E.; Lam, Lien; MacRae, Calum A.; Mohiuddin, Syed M.; Fatkin, Diane; Seidman, Christine E.; Seidman, J. G.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 29, 18.07.2017, p. 7689-7694.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of pathogenic gene mutations in LMNA and MYBPC3 that alter RNA splicing
AU - Ito, Kaoru
AU - Patel, Parth N.
AU - Gorham, Joshua M.
AU - McDonough, Barbara
AU - DePalma, Steven R.
AU - Adler, Emily E.
AU - Lam, Lien
AU - MacRae, Calum A.
AU - Mohiuddin, Syed M.
AU - Fatkin, Diane
AU - Seidman, Christine E.
AU - Seidman, J. G.
PY - 2017/7/18
Y1 - 2017/7/18
N2 - Genetic variants that cause haploinsufficiency account for many autosomal dominant (AD) disorders. Gene-based diagnosis classifies variants that alter canonical splice signals as pathogenic, but due to imperfect understanding of RNA splice signals other variants that may create or eliminate splice sites are often clinically classified as variants of unknown significance (VUS). To improve recognition of pathogenic splice-altering variants in AD disorders, we used computational tools to prioritize VUS and developed a cell-based minigene splicing assay to confirm aberrant splicing. Using this two-step procedure we evaluated all rare variants in two AD cardiomyopathy genes, lamin A/C (LMNA) and myosin binding protein C (MYBPC3). We demonstrate that 13 LMNA and 35 MYBPC3 variants identified in cardiomyopathy patients alter RNA splicing, representing a 50% increase in the numbers of established damaging splice variants in these genes. Over half of these variants are annotated as VUS by clinical diagnostic laboratories. Familial analyses of one variant, a synonymous LMNA VUS, demonstrated segregation with cardiomyopathy affection status and altered cardiac LMNA splicing. Application of this strategy should improve diagnostic accuracy and variant classification in other haploinsufficient AD disorders.
AB - Genetic variants that cause haploinsufficiency account for many autosomal dominant (AD) disorders. Gene-based diagnosis classifies variants that alter canonical splice signals as pathogenic, but due to imperfect understanding of RNA splice signals other variants that may create or eliminate splice sites are often clinically classified as variants of unknown significance (VUS). To improve recognition of pathogenic splice-altering variants in AD disorders, we used computational tools to prioritize VUS and developed a cell-based minigene splicing assay to confirm aberrant splicing. Using this two-step procedure we evaluated all rare variants in two AD cardiomyopathy genes, lamin A/C (LMNA) and myosin binding protein C (MYBPC3). We demonstrate that 13 LMNA and 35 MYBPC3 variants identified in cardiomyopathy patients alter RNA splicing, representing a 50% increase in the numbers of established damaging splice variants in these genes. Over half of these variants are annotated as VUS by clinical diagnostic laboratories. Familial analyses of one variant, a synonymous LMNA VUS, demonstrated segregation with cardiomyopathy affection status and altered cardiac LMNA splicing. Application of this strategy should improve diagnostic accuracy and variant classification in other haploinsufficient AD disorders.
UR - http://www.scopus.com/inward/record.url?scp=85024406213&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85024406213&partnerID=8YFLogxK
U2 - 10.1073/pnas.1707741114
DO - 10.1073/pnas.1707741114
M3 - Article
C2 - 28679633
AN - SCOPUS:85024406213
VL - 114
SP - 7689
EP - 7694
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 29
ER -