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Normal breast tissue of obese women is enriched for macrophage markers and macrophage-associated gene expression

  • Epidemiology
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Abstract

Activation of inflammatory pathways is one plausible mechanism underlying the association between obesity and increased breast cancer risk. However, macrophage infiltration and local biomarkers of inflammation in breast adipose tissue have seldom been studied in association with obesity. Gene expression profiles of normal breast tissue from reduction mammoplasty patients were evaluated by whole genome microarrays to identify patterns associated with obesity status (normal-weight, body mass index (BMI) <25; overweight, BMI 25–29.9; obese, BMI ≥30). The presence of macrophage-enriched inflammatory loci with immunopositivity for CD68 protein was evaluated by immunohistochemistry (IHC). After adjusting for confounding by age, 760 genes were differentially expressed (203 up and 557 down; FDR = 0.026) between normal-weight and obese women. Gene ontology analysis suggested significant enrichment for pathways involving IL-6, IL-8, CCR5 signaling in macrophages and RXRα and PPARα activation, consistent with a pro-inflammatory state and suggestive of macrophage infiltration. Gene set enrichment analysis also demonstrated that the genomic signatures of monocytes and macrophages were over-represented in the obese group with FDR of 0.08 and 0.13, respectively. Increased macrophage infiltration was confirmed by IHC, which showed that the breast adipose tissue of obese women had higher average macrophage counts (mean = 8.96 vs. 3.56 in normal-weight women) and inflammatory foci counts (mean = 4.91 vs. 2.67 in normal-weight women). Obesity is associated with local inflammation and macrophage infiltration in normal human breast adipose tissues. Given the role of macrophages in carcinogenesis, these findings have important implications for breast cancer etiology and progression.

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Acknowledgments

This project was supported by a grant from the National Institutes of Cancer (R01-CA138255), a Breast SPORE (P50CA058223) Career Development Award to M.A.T., a grant from the Avon Foundation, and the University Cancer Research Fund at the University of North Carolina. We are grateful to Melissa Rotunno for helpful comments on this study.

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Authors and Affiliations

  1. Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Xuezheng Sun & Melissa A. Troester

  2. Department of Pathology and Laboratory Medicine, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Patricia Casbas-Hernandez

  3. Program in Epidemiology and Biostatistics, Department of Public Health, University of Massachusetts at Amherst, Amherst, USA

    Carol Bigelow

  4. Department of Nutrition, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Liza Makowski

  5. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Liza Makowski & Melissa A. Troester

  6. Division of Veterinary and Animal Sciences, University of Massachusetts Amherst, Amherst, USA

    D. Joseph Jerry

  7. Pioneer Valley Life Sciences Institute, Springfield, MA, USA

    D. Joseph Jerry & Sallie Smith Schneider

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  1. Xuezheng Sun
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  2. Patricia Casbas-Hernandez
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  3. Carol Bigelow
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  4. Liza Makowski
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Correspondence to Melissa A. Troester.

Electronic supplementary material

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10549_2011_1789_MOESM1_ESM.jpg

Supplementary Fig. 1. Enrichment plots for genomic signatures of monocytes (Du et al. 2006) and macrophages (Klimcakova et al. 2011) in gene expression profile of normal breast tissues from reduction mammoplasty patients with phenotype permutation (obese vs normal-weight, n = 46). (A) Enrichment plot of the genomic profile for human peripheral blood monocytes (27 genes). (B) Enrichment plot of the genomic profile for human adipose tissue macrophage (21 genes). Detailed gene lists are shown in Supplementary Table 3. (JPEG 67.6 kb)

Supplementary Fig. 2. Representative pictures of breast adipose tissues of normal-weight patients.(PDF 429 kb)

10549_2011_1789_MOESM3_ESM.xlsx

Supplementary Table 1. The full gene list and direction of change for the genomic profile that is differentially expressed in normal breast tissue between obese and normal-weight reduction mammoplasty patients (760 genes: 203 genes up-regulated and 557 down-regulated. FDR < 0.026). (XLSX 37 kb)

10549_2011_1789_MOESM4_ESM.xlsx

Supplementary Table 2. Summary of the function analysisal of the obesity-associated signature based on the Ingenuity Pathways Analysis (IPA). (XLSX 36 kb)

10549_2011_1789_MOESM5_ESM.docx

Supplementary Table 3. Genomic signatures of human peripheral monocytes and adipose tissue macrophages adapted from Du et al. (2006) and Klimcakova et al. (2011), respectively (ref. 23 and 24), including all genes in each of these published lists that mapped to data from the reduction mammoplasty specimens on 4 × 44 K Agilent microarrays. (DOCX 19 kb)

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Sun, X., Casbas-Hernandez, P., Bigelow, C. et al. Normal breast tissue of obese women is enriched for macrophage markers and macrophage-associated gene expression. Breast Cancer Res Treat 131, 1003–1012 (2012). https://doi.org/10.1007/s10549-011-1789-3

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