Genotype and Tissue-Specific Effects on Alternative Splicing of the Transcription Factor 7-Like 2 Gene in Humans

• Published in: The journal of clinical endocrinology and metabolism 2010-03, Vol.95 (3), p.1450-1457
• Main Authors: Mondal, Ashis K, Das, Swapan K, Baldini, Giulia, Chu, Winston S, Sharma, Neeraj K, Hackney, Oksana G, Zhao, Jianhua, Grant, Struan F. A, Elbein, Steven C
• Format: Article
• Language: English
• Subjects: Adipose Tissue - metabolism; Adolescent; Adult; Alternative Splicing - genetics; Analysis of Variance; Biological and medical sciences; Diabetes Mellitus, Type 2 - genetics; Diabetes Mellitus, Type 2 - metabolism; Endocrinopathies; Exons; Feeding. Feeding behavior; Female; Fundamental and applied biological sciences. Psychology; Genetic Predisposition to Disease - genetics; Genetic Variation; Genotype; Hep G2 Cells; Humans; Insulin - metabolism; Liver - metabolism; Male; Medical sciences; Middle Aged; Muscle, Skeletal - metabolism; Organ Specificity; Original; Pancreas - metabolism; Polymorphism, Single Nucleotide - genetics; Protein Isoforms - genetics; Protein Isoforms - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; TCF Transcription Factors - genetics; TCF Transcription Factors - metabolism; Transcription Factor 7-Like 2 Protein; Vertebrates: anatomy and physiology, studies on body, several organs or systems; Vertebrates: endocrinology
• ISSN: 0021-972X; 1945-7197
• DOI: 10.1210/jc.2009-2064

Context: Noncoding single-nucleotide polymorphisms (SNPs) within the TCF7L2 gene are confirmed risk factors for type 2 diabetes, but the mechanism by which they increase risk is unknown. Objective: We hypothesized that associated SNPs alter TCF7L2 splicing and that splice forms have altered biological roles. Design: Splice forms and 5′ and 3′ untranslated regions were characterized in sc adipose, muscle, liver, HepG2 cells, pancreas, and islet. Isoform-specific transcript levels were quantified in sc adipose. Alternative splice forms were characterized in HepG2 liver cells under glucose and insulin conditions and in SGBS cells with differentiation. Major isoforms were characterized by transfection. Setting: The study was conducted at an ambulatory general clinical research center. Patients: Patients included 78 healthy, nondiabetic study subjects characterized for insulin sensitivity and secretion. Results: We identified 32 alternatively spliced transcripts and multiple-length 3′ untranslated region transcripts in adipose, muscle, islet, and pancreas. Alternative exons 3a, 12, 13, and 13a were observed in all tissues, whereas exon 13b was islet specific. Transcripts retaining exons 13 and 13a but not total TCF7L2 transcripts were significantly correlated with both obesity measures (P < 0.01) and rs7903146 genotype (P < 0.026) in sc adipose. Insulin (5–10 nm) suppressed all TCF7L2 isoforms in SGBS cells but suppressed exon 13a-containing isoforms most significantly (P < 0.001). The isoform distribution differed throughout SGBS cell differentiation. Isoforms with predicted early stop codons yielded stable proteins of the predicted size, bound β-catenin, and targeted correctly to the nucleus. Conclusions: Intronic TCF7L2 variants may regulate alternative transcript isoforms, which in turn may have distinct physiologic roles. Intronic TCF7L2 variants associated with type 2 diabetes may regulate expression of alternative transcript isoforms, which in turn likely have distinct physiologic roles.