FTO Obesity Variant Circuitry and Adipocyte Browning in Humans.
BACKGROUND
Genomewide association studies can be used to identify disease-relevant genomic regions, but interpretation of the data is challenging. The FTO region harbors the strongest genetic association with obesity, yet the mechanistic basis of this association remains elusive.


METHODS
We examined epigenomic data, allelic activity, motif conservation, regulator expression, and gene coexpression patterns, with the aim of dissecting the regulatory circuitry and mechanistic basis of the association between the FTO region and obesity. We validated our predictions with the use of directed perturbations in samples from patients and from mice and with endogenous CRISPR-Cas9 genome editing in samples from patients.


RESULTS
Our data indicate that the FTO allele associated with obesity represses mitochondrial thermogenesis in adipocyte precursor cells in a tissue-autonomous manner. The rs1421085 T-to-C single-nucleotide variant disrupts a conserved motif for the ARID5B repressor, which leads to derepression of a potent preadipocyte enhancer and a doubling of IRX3 and IRX5 expression during early adipocyte differentiation. This results in a cell-autonomous developmental shift from energy-dissipating beige (brite) adipocytes to energy-storing white adipocytes, with a reduction in mitochondrial thermogenesis by a factor of 5, as well as an increase in lipid storage. Inhibition of Irx3 in adipose tissue in mice reduced body weight and increased energy dissipation without a change in physical activity or appetite. Knockdown of IRX3 or IRX5 in primary adipocytes from participants with the risk allele restored thermogenesis, increasing it by a factor of 7, and overexpression of these genes had the opposite effect in adipocytes from nonrisk-allele carriers. Repair of the ARID5B motif by CRISPR-Cas9 editing of rs1421085 in primary adipocytes from a patient with the risk allele restored IRX3 and IRX5 repression, activated browning expression programs, and restored thermogenesis, increasing it by a factor of 7.


CONCLUSIONS
Our results point to a pathway for adipocyte thermogenesis regulation involving ARID5B, rs1421085, IRX3, and IRX5, which, when manipulated, had pronounced pro-obesity and anti-obesity effects. (Funded by the German Research Center for Environmental Health and others.).
From Beth Israel Deaconess Medical Center and Hebrew SeniorLife, Gerontology Division, Harvard Medical School, Boston (M.C., Y.-H.H.); Massachusetts Institute of Technology (MIT) Computer Science and Artificial Intelligence Laboratory (M.C., G.Q., W.M., N.A.A., M.K.), and Broad Institute of MIT and Harvard, Cambridge (M.C., G.Q., W.M., M.K.); Clinical Cooperation Group "Nutrigenomics and Type 2 Diabetes," Helmholtz Center Munich (M.C., H.H.), and Else Kröner-Fresenius Center for Nutritional Medicine, Klinikum rechts der Isar, ZIEL-Institute for Food and Health, Technische Universität München (M.C., V.G., I.S.S., H.H.), Munich, Germany; KG Jebsen Center for Diabetes Research, Department of Clinical Science, University of Bergen, and Hormone Laboratory, Haukeland University Hospital, Bergen, Norway (S.N.D., C.H., G.M.); Program in Developmental and Stem Cell Biology, Hospital for Sick Children, and Department of Molecular Genetics, University of Toronto (K.-H.K., V.P., J.L., C.-C.H.), and Department of Medicine, Lunenfeld-Tanenbaum Research Institute, Mt. Sinai Hospital (J.L.B., D.J.D.), Toronto; and the Department of Molecular and Clinical Medicine, Institute of Medicine, University of Gothenburg, Gothenburg, Sweden (P.-A.S.).
Melina
M
Claussnitzer
Simon N
SN
Dankel
Kyoung-Han
KH
Kim
Gerald
G
Quon
Wouter
W
Meuleman
Christine
C
Haugen
Viktoria
V
Glunk
Isabel S
IS
Sousa
Jacqueline L
JL
Beaudry
Vijitha
V
Puviindran
Nezar A
NA
Abdennur
Jannel
J
Liu
Per-Arne
PA
Svensson
Yi-Hsiang
YH
Hsu
Daniel J
DJ
Drucker
Gunnar
G
Mellgren
Chi-Chung
CC
Hui
Hans
H
Hauner
Manolis
M
Kellis
D017667:Adipocytes / Q000378:metabolism*; D000483:Alleles; D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; D000818:Animals; D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D057890:Epigenomics; D015870:Gene Expression; D005818:Genetic Engineering; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D009765:Obesity / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D011506:Proteins / Q000235:genetics*; D017393:RNA Editing; D012306:Risk; D022722:Thermogenesis / Q000235:genetics* / Q000502:physiology
D011506:Proteins; D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; C526492:FTO protein, human