Details

Autor(en) / Beteiligte

• Majithia, Amit R
• Tsuda, Ben
• Agostini, Maura
• Gnanapradeepan, Keerthana
• Rice, Robert
• Peloso, Gina
• Patel, Kashyap A
• Zhang, Xiaolan
• Broekema, Marjoleine F
• Patterson, Nick
• Duby, Marc
• Sharpe, Ted
• Kalkhoven, Eric
• Rosen, Evan D
• Barroso, Inês
• Ellard, Sian
• Kathiresan, Sekar
• O'Rahilly, Stephen
• Chatterjee, Krishna
• Florez, Jose C
• Mikkelsen, Tarjei
• Savage, David B
• Altshuler, David

Titel

Prospective functional classification of all possible missense variants in PPARG

Ist Teil von

• Nature genetics, 2016-12, Vol.48 (12), p.1570-1575

Ort / Verlag

United States: Nature Publishing Group

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• Clinical exome sequencing routinely identifies missense variants in disease-related genes, but functional characterization is rarely undertaken, leading to diagnostic uncertainty. For example, mutations in PPARG cause Mendelian lipodystrophy and increase risk of type 2 diabetes (T2D). Although approximately 1 in 500 people harbor missense variants in PPARG, most are of unknown consequence. To prospectively characterize PPARγ variants, we used highly parallel oligonucleotide synthesis to construct a library encoding all 9,595 possible single-amino acid substitutions. We developed a pooled functional assay in human macrophages, experimentally evaluated all protein variants, and used the experimental data to train a variant classifier by supervised machine learning. When applied to 55 new missense variants identified in population-based and clinical sequencing, the classifier annotated 6 variants as pathogenic; these were subsequently validated by single-variant assays. Saturation mutagenesis and prospective experimental characterization can support immediate diagnostic interpretation of newly discovered missense variants in disease-related genes.