title: In vivo severity ranking of Ras pathway mutations associated with developmental disorders.
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.
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544.|Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.|Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Granton A
authors.identifier: 0000-0003-2803-9831
authors.initials: GA
authors.lastname: Jindal
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544.|Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.
authors.forename: Yogesh
authors.initials: Y
authors.lastname: Goyal
authors.affiliation: Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.|Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Kei
authors.initials: K
authors.lastname: Yamaya
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544.|Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.
authors.forename: Alan S
authors.initials: AS
authors.lastname: Futran
authors.affiliation: Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.
authors.forename: Iason
authors.initials: I
authors.lastname: Kountouridis
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Courtney A
authors.initials: CA
authors.lastname: Balgobin
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Trudi
authors.initials: T
authors.lastname: Schüpbach
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Rebecca D
authors.initials: RD
authors.lastname: Burdine
authors.affiliation: Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544; stas@princeton.edu.|Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544.|Department of Molecular Biology, Princeton University, Princeton, NJ 08544.
authors.forename: Stanislav Y
authors.initials: SY
authors.lastname: Shvartsman
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002658:Developmental Disabilities / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics; D006801:Humans; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000235:genetics; D009154:Mutation; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D015027:Zebrafish / Q000196:embryology / Q000235:genetics; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018631:ras Proteins / Q000235:genetics*
chemical_list: D029721:Drosophila Proteins; D047428:Protein Kinase Inhibitors; D029961:Zebrafish Proteins; D048369:MAP Kinase Kinase 1; D018631:ras Proteins
keywords: Drosophila; MEK inhibitor; MEK1; RASopathies; zebrafish