In vivo severity ranking of Ras pathway mutations associated with developmental disorders. Author Granton Jindal, Yogesh Goyal, Kei Yamaya, Alan Futran, Iason Kountouridis, Courtney Balgobin, Trudi Schüpbach, Rebecca Burdine, Stanislav Shvartsman Publication Year 2017 Type Journal Article Abstract Germ-line mutations in components of the Ras/MAPK pathway result in developmental disorders called RASopathies, affecting about 1/1,000 human births. Rapid advances in genome sequencing make it possible to identify multiple disease-related mutations, but there is currently no systematic framework for translating this information into patient-specific predictions of disease progression. As a first step toward addressing this issue, we developed a quantitative, inexpensive, and rapid framework that relies on the early zebrafish embryo to assess mutational effects on a common scale. Using this assay, we assessed 16 mutations reported in MEK1, a MAPK kinase, and provide a robust ranking of these mutations. We find that mutations found in cancer are more severe than those found in both RASopathies and cancer, which, in turn, are generally more severe than those found only in RASopathies. Moreover, this rank is conserved in other zebrafish embryonic assays and Drosophila-specific embryonic and adult assays, suggesting that our ranking reflects the intrinsic property of the mutant molecule. Furthermore, this rank is predictive of the drug dose needed to correct the defects. This assay can be readily used to test the strengths of existing and newly found mutations in MEK1 and other pathway components, providing the first step in the development of rational guidelines for patient-specific diagnostics and treatment of RASopathies. Keywords Animals, Drosophila Proteins, Dose-Response Relationship, Drug, Humans, Mutation, Phenotype, Animals, Genetically Modified, Drosophila melanogaster, MAP Kinase Signaling System, ras Proteins, Zebrafish, Zebrafish Proteins, Protein Kinase Inhibitors, Developmental Disabilities, MAP Kinase Kinase 1 Journal Proc Natl Acad Sci U S A Volume 114 Issue 3 Pages 510-515 Date Published 2017 Jan 17 ISSN Number 1091-6490 DOI 10.1073/pnas.1615651114 Alternate Journal Proc Natl Acad Sci U S A PMCID PMC5255624 PMID 28049852 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML