Identification and functional characterization of a missense mutation in resistin in two patients with severe obesity and insulin resistance
| Autor: | Wim Van Hul, Frida Peiffer, Doreen Zegers, Ilse Mertens, Fenna de Freitas, Kristine Desager, Luc Van Gaal, Sigri Beckers, Armand V. Peeters, Guy Massa, Stijn Verhulst, An Verrijken |
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| Rok vydání: | 2011 |
| Předmět: |
Adult
Male Untranslated region Proband Heterozygote medicine.medical_specialty Adolescent Endocrinology Diabetes and Metabolism DNA Mutational Analysis Genetic Vectors Green Fluorescent Proteins Mutant Mutation Missense Adipokine Enzyme-Linked Immunosorbent Assay Transfection Body Mass Index Mice Young Adult Endocrinology Mutant protein 3T3-L1 Cells Internal medicine Animals Humans Medicine Missense mutation Family Resistin RNA Messenger Child 3' Untranslated Regions business.industry Body Weight Exons General Medicine Middle Aged Obesity Morbid Mutation testing Female Human medicine Insulin Resistance business |
| Zdroj: | European journal of endocrinology |
| ISSN: | 1479-683X 0804-4643 |
| Popis: | ObjectiveIn this study, we hypothesized that mutations in the resistin encoding gene, RETN, may cause a monogenic form of obesity.Design/methodsWe screened the coding region of RETN in 81 morbidly obese adults, 263 overweight and obese children/adolescents, and 116 healthy lean subjects. In vitro experiments include qPCR, ELISA, and western blot for WT and mutant resistin transfected into 3T3-L1 adipocytes.ResultsMutation analysis identified five sequence variants in our patient populations: 3′-UTR +87 G/A, 3′-UTR +100 A/G, T73T, IV3-61 C/A, and C78S. In our control population, we only found the 3′-UTR +87 G/A variant. We started functional experiments for the C78S mutation that was found in a 20-year-old obese male (body mass index (BMI)=39.7 kg/m2) and his obese mother (BMI=31.9 kg/m2). In vitro testing demonstrated that the mutation does not impair mRNA expression. We identified a 100-fold lower extracellular protein concentration for mutant resistin compared with WT levels using a resistin ELISA on cell culture medium (P=4.87×10−6). We also detected a decreased intracellular concentration for the mutant protein (tenfold lower relative levels, P=0.007). The plasma resistin levels of the proband and his mother, however, did not differ significantly from lean control individuals.ConclusionsIn conclusion, we identified the first missense mutation in resistin in a morbidly obese proband and his obese mother. Functional testing of the mutant protein suggests that the C78S mutant protein is degraded, possibly resulting in a decreased extracellular concentration, which may predispose to obesity. |
| Databáze: | OpenAIRE |
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