DNA methylation and gene expression of HIF3A: cross-tissue validation and associations with BMI and insulin resistance.
Associations between BMI and DNA methylation of hypoxia-inducible factor 3-alpha (HIF3A) in both blood cells and subcutaneous adipose tissue (SAT) have been reported. In this study, we investigated associations between BMI and HIF3A DNA methylation in the blood and SAT from the same individuals, and whether HIF3A gene expression in SAT and skeletal muscle biopsies showed associations with BMI and insulin resistance. Furthermore, we aimed to investigate gender specificity and heritability of these traits.



We studied 137 first-degree relatives of type 2 diabetes (T2D) patients from 48 families, from whom we had SAT and muscle biopsies. DNA methylation of four CpG sites in the HIF3A promoter was analyzed in the blood and SAT by pyrosequencing, and HIF3A gene expression was analyzed in SAT and muscle by qPCR. An index of whole-body insulin sensitivity was estimated from oral glucose tolerance tests.



BMI was associated with HIF3A methylation at one CpG site in the blood, and there was a positive association between the blood and SAT methylation levels at a different CpG site within the individuals. The SAT methylation level did not correlate with HIF3A gene expression. Interestingly, HIF3A expression in SAT, but not in muscle, associated negatively with BMI and whole-body insulin resistance. We found a significant effect of familiality on HIF3A methylation levels in the blood and HIF3A expression levels in skeletal muscle.



Our findings are in line with the previously reported link between BMI and DNA methylation of HIF3A in the blood. The tissue-specific results of HIF3A gene expression indicate that SAT is the more functional tissue in which a low expression may adversely affect whole-body insulin sensitivity.
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.
Ailsa Maria
AM
Main
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.|Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Linn
L
Gillberg
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.
Anna Louisa
AL
Jacobsen
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.|Department of Clinical Sciences, Lund University Diabetes Centre, Malmö, Sweden.
Emma
E
Nilsson
Section of Metabolic Genetics, The Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Anette Prior
AP
Gjesing
Section of Metabolic Genetics, The Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Torben
T
Hansen
Section of Metabolic Genetics, The Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Oluf
O
Pedersen
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.|The Danish Diabetes Academy, Odense University Hospital, Odense, Denmark.
Rasmus
R
Ribel-Madsen
Department of Endocrinology, Rigshospitalet, Section 7652, Tagensvej 20, DK-2200 Copenhagen, Denmark.|Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Allan
A
Vaag
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D051017:Apoptosis Regulatory Proteins; D051792:Basic Helix-Loop-Helix Proteins / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D015992:Body Mass Index; D018899:CpG Islands; D019175:DNA Methylation; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D044127:Epigenesis, Genetic; D005260:Female; D006801:Humans; D007333:Insulin Resistance; D008297:Male; D008875:Middle Aged; D009928:Organ Specificity; D010375:Pedigree; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins; D017422:Sequence Analysis, DNA / Q000379:methods*; D050151:Subcutaneous Fat / Q000378:metabolism*
D051017:Apoptosis Regulatory Proteins; D051792:Basic Helix-Loop-Helix Proteins; D012097:Repressor Proteins; C116973:HIF3A protein, human
Epigenetics; Heritability; Insulin sensitivity; Obesity; Type 2 diabetes