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04.05.2020 | Original Article

MLH1 intronic variants mapping to + 5 position of splice donor sites lead to deleterious effects on RNA splicing

verfasst von: Tamara Alejandra Piñero, Omar Soukarieh, Marion Rolain, Karin Alvarez, Francisco López-Köstner, Giovana Tardin Torrezan, Dirce Maria Carraro, Ivana Lucia De Oliveira Nascimento, Thaís Ferreira Bomfim, Taísa Manuela Bonfim Machado-Lopes, Juliana Côrtes Freitas, Maria Betânia Toralles, Kiyoko Abe Sandes, Benedito Mauro Rossi, Samuel Aguiar Junior, Joanna Meira, Mev Dominguez-Valentin, Pål Møller, Carlos Alberto Vaccaro, Alexandra Martins, Walter Hernán Pavicic

Erschienen in: Familial Cancer | Ausgabe 4/2020

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Abstract

Germline pathogenic variants in the DNA mismatch repair genes (MMR): MLH1, MSH2, MSH6, and PMS2, are causative of Lynch syndrome (LS). However, many of the variants mapping outside the invariant splice site positions (IVS ± 1, IVS ± 2) are classified as variants of unknown significance (VUS). Three such variants (MLH1 c.588+5G>C, c.588+5G>T and c.677+5G>A) were identified in 8 unrelated LS families from Argentina, Brazil and Chile. Herein, we collected clinical information on these families and performed segregation analysis and RNA splicing studies to assess the implication of these VUS in LS etiology. Pedigrees showed a clear pattern of variant co-segregation with colorectal cancer and/or other LS-associated malignancies. Tumors presented deficient expression of MLH1-PMS2 proteins in 7/7 of the LS families, and MSI-high status in 3/3 cases. Moreover, RNA analyses revealed that c.588+5G>C and c.588+5G>T induce skipping of exon 7 whereas c.677+5G>A causes skipping of exon 8. In sum, we report that the combined clinical findings in the families and the molecular studies provided the evidences needed to demonstrate that the three MLH1 variants are causative of LS and to classify c.588+5G>C and c.677+5G>A as class 5 (pathogenic), and c.588+5G>T as class 4 (likely-pathogenic). Our findings underline the importance of performing clinical and family analyses, as well as RNA splicing assays in order to determine the clinical significance of intronic variants, and contribute to the genetic counseling and clinical management of patients and their relatives.

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Titel: MLH1 intronic variants mapping to + 5 position of splice donor sites lead to deleterious effects on RNA splicing
verfasst von: Tamara Alejandra Piñero
Omar Soukarieh
Marion Rolain
Karin Alvarez
Francisco López-Köstner
Giovana Tardin Torrezan
Dirce Maria Carraro
Ivana Lucia De Oliveira Nascimento
Thaís Ferreira Bomfim
Taísa Manuela Bonfim Machado-Lopes
Juliana Côrtes Freitas
Maria Betânia Toralles
Kiyoko Abe Sandes
Benedito Mauro Rossi
Samuel Aguiar Junior
Joanna Meira
Mev Dominguez-Valentin
Pål Møller
Carlos Alberto Vaccaro
Alexandra Martins
Walter Hernán Pavicic
Publikationsdatum: 04.05.2020
Verlag: Springer Netherlands
Erschienen in: Familial Cancer / Ausgabe 4/2020
Print ISSN: 1389-9600
Elektronische ISSN: 1573-7292
DOI: https://doi.org/10.1007/s10689-020-00182-5

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MLH1 intronic variants mapping to + 5 position of splice donor sites lead to deleterious effects on RNA splicing

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