No Association between FTO or HHEX and Endometrial Cancer Risk

Obesity and type II diabetes are major risk factors for endometrial cancer. Genetic variation that is associated with obesity and diabetes might provide clues to the molecular pathways mediating endometrial carcinogenesis. A genome-wide association study of body mass index (BMI) identified a 47 kb region on chromosome 16 encompassing the FTO gene intron 1-exon 2-intron 2 that is marked by tag-single nucleotide polymorphisms (SNP) rs8050136 (and the correlated SNP rs9939609; r² = 1) to be associated with BMI (1). Another genome-wide association study of type II diabetes (2) identified the HHEX gene region on chromosome 10 marked by rs1111875 that is also associated with BMI. To examine whether genetic variants of FTO and HHEX are associated with endometrial cancer risk, we genotyped tagSNPs and candidate SNPs in the Polish Endometrial Case-Control Study (PECS) of 417 endometrial cancer cases and 406 population-based controls (3). Significant findings in this set were then selected for validation in a replication set of approximately 2,347 cases and 3,140 controls from three additional studies.

Genotyping of 189 tagSNPs in FTO (including rs8050136) and 5 tagSNPs in HHEX (including rs1111875) were done as part of an Illumina Infinium custom iSelect chip using a SNP selection strategy described previously (4). For FTO, four SNPs were excluded due to violations of quality control measures: concordance of 1% replicates, completion proportions, and departure from Hardy-Weinberg proportions (P < 0.05). An additional 14 SNPs were excluded due to minor allele frequencies (MAF) of <0.05 among controls. For HHEX, all five SNPs passed quality control filters. Odds ratios (OR) and 95% confidence intervals (CI) were calculated to estimate the association between SNPs and endometrial cancer risk using unconditional logistic regression models, controlling for matching factors, age, and study site. We also conducted analyses of haplotypes, including the sequential haplotype scan (5) and the variable-sized sliding window–regularized regression association analysis (6) to localize a set of adjacent markers associated with risk. Due to the size of the FTO gene, the sequential haplotype scan was performed in three overlapping sections [section 1 (SNPs 1-65), rs8055834-rs17820875; section 2 (SNPs 60-115), rs10521303-rs8056199; and section 3 (SNPs 110-171), rs16952730-rs16953089].

Among PECS controls, carrying an increased number of copies of the minor A allele of rs8050136 in FTO was associated with increased mean BMI (Kruskal-Wallis P = 0.015) but not with the prevalence of diabetes (χ², P = 0.26). rs1111875 in HHEX was not associated with BMI (P = 0.16) or diabetes (P = 0.56). In the PECS case-control analyses, the minor A allele of rs8050136 was not associated with endometrial cancer risk (per allele OR, 1.05; 95% CI, 0.86-1.28; P = 0.64). However, 20 of the remaining 171 FTO tagSNPs were significantly associated with endometrial cancer risk (per allele P values ranged from 0.00068 to 0.027) and represented independent SNPs (n = 4) or clustered into three linkage disequilibrium blocks. Haplotype analyses identified strong signals (haplotype P < 10⁻³) in two of these regions (Figs. 1 and 2). The first region resides in intron 4 and is marked by SNP rs8063241 (OR, 0.71; 95% CI, 0.56-0.88, P = 1.7 × 10⁻³; Fig. 1). The second region in intron 8 is marked by three correlated tagSNPs that also had the lowest P values in the single locus analysis [rs2689264 (MAF = 0.17): OR, 1.57; 95% CI, 1.22-2.02; P = 4.5 × 10⁻⁴; rs12927155 (MAF = 0.17): OR, 1.56; 95% CI, 1.21-2.01; P = 5.8 × 10⁻⁴; rs2540776 (MAF = 0.17): OR, 1.54; 95% CI, 1.19-1.99; P = 8.7 × 10⁻⁴; Fig. 2]. The candidate SNP in HHEX, rs1111875, was associated with a 20% lower risk of endometrial cancer for each minor T allele (per allele OR, 0.80; 95% CI, 0.66-0.97; P = 0.026). No other HHEX loci were associated with risk.

In an attempt to replicate our findings for FTO SNP rs12927155 and HHEX SNP rs1111875, we approached three independent case-control studies of women of European ancestry (5,522 subjects; Table 1), including the Study of Epidemiology and Risk Factors in Cancer Heredity (SEARCH) with 1,494 endometrial cancer cases and 1,600 community controls, the Australian National Endometrial Cancer Study (ANECS) with 1,048 endometrial cancer cases and 1,010 population-based controls (7), and the Leuven Endometrial Cancer Study (LES; ref. 8) with 206 endometrial cancer cases and 649 hospital-based controls. The distribution of age and BMI were similar for the three studies [age range (median): SEARCH, 26-71 (56); ANECS, 26-80 (62); LES, 20-80 (48); and BMI range (median): SEARCH, not reported; ANECS, 15.1-67.3 (28.0); LES, 16.4-89.0 (24.9)] and with the PECS. We excluded controls with a history of hysterectomy (including 249 for SEARCH, 95 for ANECS, and 1 for LES). The SEARCH samples were genotyped using TaqMan assays, ANECS and LES samples were genotyped using the Sequenom iPLEX platform.

Among these three studies, the pooled per allele ORs, adjusted for continuous age and study for FTO was rs12927155 (OR, 0.94; 95% CI, 0.83-1.06; P = 0.29) and for HHEX, it was rs1111875 (OR, 1.00; 95% CI, 0.92-1.10; P = 0.96; Table 1). Between-study heterogeneity was not evident among these studies (P = 0.23 and 0.74, respectively), and the CIs for both SNPs excluded the ORs from the PECS.

Our data indicate that common genetic variants in two genes previously related to obesity (FTO) and diabetes (HHEX) by genome-wide association scans were not associated with endometrial cancer risk.

No potential conflicts of interest were disclosed.

For the PECS, the authors thank Neonila Szeszenia-Dabrowska of the Nofer Institute of Occupational Medicine (Lodz, Poland) and Witold Zatonski of the M. Sklodowska-Curie Institute of Oncology and Cancer Center (Warsaw, Poland) for their contribution to the Polish Endometrial Cancer Study. Anita Soni (Westat, Rockville, MD) and Pei Chao (IMS, Silver Spring, MD) have been invaluable to the management of the study. This work would not be possible without the dedicated efforts of the physicians, nurses, interviewers, and study participants. The authors thank all the women who participated in the SEARCH study as well as Caroline Baynes, Don Conroy, Craig Luccarini, and Mitul Shah for work within the SEARCH study. The ANECS Group comprises: A.B. Spurdle, P. Webb, J. Young (Queensland Institute of Medical Research); Consumer representative: L. McQuire; Clinical Collaborators: NSW: S. Baron-Hay, D. Bell, A. Bonaventura, A. Brand, S. Braye, J. Carter, F. Chan, C. Dalrymple, A. Ferrier (deceased), G. Gard, N. Hacker, R. Hogg, R. Houghton, D. Marsden, K. McIlroy, G. Otton, S. Pather, A. Proietto, G. Robertson, J. Scurry, R. Sharma, G. Wain, F. Wong; Qld: J. Armes, A. Crandon, M. Cummings, R. Land, J. Nicklin, L. Perrin, A. Obermair, B. Ward; SA: M. Davy, T. Dodd, J. Miller, M. Oehler, S. Paramasivum, J. Pierides, F. Whitehead; Tas: P. Blomfield, D. Challis; Vic: D. Neesham, J. Pyman, M. Quinn, R. Rome, M. Weitzer; WA: B. Brennan, I. Hammond, Y. Leung, A. McCartney, C. Stewart, J. Thompson; Project Managers: S. O'Brien, S. Moore; Laboratory Manager: K. Ferguson; Pathology Support: M. Walsh; Admin Support: R. Cicero, L. Green, J. Griffith, L. Jackman, B. Ranieri; Laboratory Assistants: M. O'Brien, P. Schultz; Research Nurses: B. Alexander, C. Baxter, H. Croy, A. Fitzgerald, E. Herron, C. Hill, M. Jones, J. Maidens, A. Marshall, K. Martin, J. Mayhew, E. Minehan, D. Roffe, H. Shirley, H. Steane, A. Stenlake, A. Ward, S. Webb, J. White. ANECS would also like to gratefully acknowledge the cooperation of the following institutions: NSW: John Hunter Hospital, Liverpool Hospital, Mater Misericordiae Hospital (Sydney), Mater Misericordiae Hospital (Newcastle), Newcastle Private Hospital, North Shore Private Hospital, Royal Hospital for Women, Royal Prince Alfred Hospital, Royal North Shore Hospital, Royal Prince Alfred Hospital, St. George Hospital, Westmead Hospital, Westmead Private Hospital; Qld: Brisbane Private Hospital, Greenslopes Hospital, Mater Misericordiae Hospitals, Royal Brisbane and Women's Hospital, Wesley Hospital, Queensland Cancer Registry; SA: Adelaide Pathology Partners, Burnside Hospital, Calvary Hospital, Flinders Medical Centre, Queen Elizabeth Hospital, Royal Adelaide Hospital, South Australian Cancer Registry; Tas: Launceston Hospital, North West Regional Hospitals, Royal Hobart Hospital; Vic: Freemasons Hospital, Melbourne Pathology Services, Mercy Hospital for Women, Royal Women's Hospital, Victorian Cancer Registry; WA: King Edward Memorial Hospital, St. John of God Hospitals Subiaco & Murdoch, Western Australian Cancer Registry. We also thank Felicity Lose, Jyotsna Batra, Xiaoqing Chen, and Jonathan Beesley from The Molecular Cancer Epidemiology and Cancer Genetic laboratories at QIMR for technical assistance. We also thank the Australian Red Cross Blood Services (ARCBS) donors who participated as healthy controls in this study. We are grateful to the staff at ARCBS for their assistance with the collection of risk factor information and blood samples, and Melanie Higgins, Kimberley Hinze, Felicity Lose, and members of the Molecular Cancer Epidemiology Laboratory for their assistance with collection and processing of blood samples. The Leuven Endometrial Cancer Study gratefully acknowledges the contribution of Drs. Evelyn Despierre and Gilian Peuteman for assistance.

Grant Support: PECS was funded by the intramural research program at the National Cancer Institute, Division of Cancer Epidemiology and Genetics in the Hormonal and Reproductive Epidemiology Branch. Analysis and conduct of the SEARCH study was funded by Cancer Research UK grants (C20/A3084, C1287/A10118, C490/A11021, C8197/A10123, C1287/A7497). Cancer Research UK grant (C8197/A10865; A.M. Dunning). ANECS was supported by a project grants from the National Health and Medical Research Council (NHMRC) of Australia (ID no. 339435), the Cancer Council Queensland (ID no. 4196615) and Cancer Council Tasmania (ID no. 403031 and ID no. 457636). NHMRC Senior Research Fellowship (A. Spurdle). Australian Postgraduate Award, an Institute of Health and Biomedical Innovation Ph.D. Top-Up and a Smart State Ph.D. Award (T. O'Mara). The Leuven Endometrial Cancer Study was supported by the Verelst Foundation for Endometrial Cancer.

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