CDKN2A Unclassified Variants in Familial Malignant Melanoma: Combining Functional and Computational Approaches for Their Assessment

Maria Chiara Scaini, Giovanni Minervini, Lisa Elefanti, Paola Ghiorzo, Lorenza Pastorino, Silvia Tognazzo, Simona Agata, Monica Quaggio, Daniela Zullato, Giovanna Bianchi-Scarrà, Marco Montagna, Emma D'Andrea, Chiara Menin, Silvio C E Tosatto

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

CDKN2A codes for two oncosuppressors by alternative splicing of two first exons: p16INK4a and p14ARF. Germline mutations are found in about 40% of melanoma-prone families, and most of them are missense mutations mainly affecting p16INK4a. A growing number of p16INK4a variants of uncertain significance (VUS) are being identified but, unless their pathogenic role can be demonstrated, they cannot be used for identification of carriers at risk. Predicting the effect of these VUS by either a "standard" in silico approach, or functional tests alone, is rather difficult. Here, we report a protocol for the assessment of any p16INK4a VUS, which combines experimental and computational tools in an integrated approach. We analyzed p16INK4a VUS from melanoma patients as well as variants derived through permutation of conserved p16INK4a amino acids. Variants were expressed in a p16INK4a-null cell line (U2-OS) and tested for their ability to block proliferation. In parallel, these VUS underwent in silico prediction analysis and molecular dynamics simulations. Evaluation of in silico and functional data disclosed a high agreement for 15/16 missense mutations, suggesting that this approach could represent a pilot study for the definition of a protocol applicable to VUS in general, involved in other diseases, as well.

Original languageEnglish
Pages (from-to)828-840
Number of pages13
JournalHuman Mutation
Volume35
Issue number7
DOIs
Publication statusPublished - 2014

Fingerprint

Computer Simulation
Melanoma
Missense Mutation
Tumor Suppressor Protein p14ARF
Null Lymphocytes
Aptitude
Germ-Line Mutation
Alternative Splicing
Molecular Dynamics Simulation
Exons
Amino Acids
Cell Line
Cutaneous Malignant Melanoma

Keywords

  • CDKN2A
  • Familial melanoma
  • P16INK4a
  • Variants of uncertain significance
  • VUS

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Medicine(all)

Cite this

CDKN2A Unclassified Variants in Familial Malignant Melanoma : Combining Functional and Computational Approaches for Their Assessment. / Scaini, Maria Chiara; Minervini, Giovanni; Elefanti, Lisa; Ghiorzo, Paola; Pastorino, Lorenza; Tognazzo, Silvia; Agata, Simona; Quaggio, Monica; Zullato, Daniela; Bianchi-Scarrà, Giovanna; Montagna, Marco; D'Andrea, Emma; Menin, Chiara; Tosatto, Silvio C E.

In: Human Mutation, Vol. 35, No. 7, 2014, p. 828-840.

Research output: Contribution to journalArticle

Scaini, MC, Minervini, G, Elefanti, L, Ghiorzo, P, Pastorino, L, Tognazzo, S, Agata, S, Quaggio, M, Zullato, D, Bianchi-Scarrà, G, Montagna, M, D'Andrea, E, Menin, C & Tosatto, SCE 2014, 'CDKN2A Unclassified Variants in Familial Malignant Melanoma: Combining Functional and Computational Approaches for Their Assessment', Human Mutation, vol. 35, no. 7, pp. 828-840. https://doi.org/10.1002/humu.22550
Scaini, Maria Chiara ; Minervini, Giovanni ; Elefanti, Lisa ; Ghiorzo, Paola ; Pastorino, Lorenza ; Tognazzo, Silvia ; Agata, Simona ; Quaggio, Monica ; Zullato, Daniela ; Bianchi-Scarrà, Giovanna ; Montagna, Marco ; D'Andrea, Emma ; Menin, Chiara ; Tosatto, Silvio C E. / CDKN2A Unclassified Variants in Familial Malignant Melanoma : Combining Functional and Computational Approaches for Their Assessment. In: Human Mutation. 2014 ; Vol. 35, No. 7. pp. 828-840.
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