title: Intermittent energy restriction induces changes in breast gene expression and systemic metabolism.
abstract: Observational studies suggest weight loss and energy restriction reduce breast cancer risk. Intermittent energy restriction (IER) reduces weight to the same extent as, or more than equivalent continuous energy restriction (CER) but the effects of IER on normal breast tissue and systemic metabolism as indicators of breast cancer risk are unknown.
  
  
  
  We assessed the effect of IER (two days of 65 % energy restriction per week) for one menstrual cycle on breast tissue gene expression using Affymetrix GeneChips, adipocyte size by morphometry, and systemic metabolism (insulin resistance, lipids, serum and urine metabolites, lymphocyte gene expression) in 23 overweight premenopausal women at high risk of breast cancer. Unsupervised and supervised analyses of matched pre and post IER biopsies in 20 subjects were performed, whilst liquid and gas chromatography mass spectrometry assessed corresponding changes in serum and urine metabolites in all subjects after the two restricted and five unrestricted days of the IER.
  
  
  
  Women lost 4.8 % (±2.0 %) of body weight and 8.0 % (±5.0 %) of total body fat. Insulin resistance (homeostatic model assessment (HOMA)) reduced by 29.8 % (±17.8 %) on the restricted days and by 11 % (±34 %) on the unrestricted days of the IER. Five hundred and twenty-seven metabolites significantly increased or decreased during the two restricted days of IER. Ninety-one percent of these returned to baseline after 5 days of normal eating. Eleven subjects (55 %) displayed reductions in energy restriction-associated metabolic gene pathways including lipid synthesis, gluconeogenesis and glycogen synthesis. Some of these women also had increases in genes associated with breast epithelial cell differentiation (secretoglobulins, milk proteins and mucins) and decreased collagen synthesis (TNMD, PCOLCE2, TIMP4). There was no appreciable effect of IER on breast gene expression in the other nine subjects. These groups did not differ in the degree of changes in weight, total body fat, fat cell size or serum or urine metabolomic markers. Corresponding gene changes were not seen in peripheral blood lymphocytes.
  
  
  
  The transcriptional response to IER is variable in breast tissue, which was not reflected in the systemic response, which occurred in all subjects. The mechanisms of breast responsiveness/non-responsiveness require further investigation.
  
  
  
  ISRCTN77916487 31/07/2012.
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK. michelle.harvie@manchester.ac.uk.
authors.forename: Michelle N
authors.initials: MN
authors.lastname: Harvie
authors.affiliation: Applied Bioinformatics of Cancer, University of Edinburgh, Cancer Research UK Centre, Institute of Genetics and Molecular Medicine, Carrington Crescent, Edinburgh, EH4 2XR, UK.
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Sims
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK.
authors.forename: Mary
authors.initials: M
authors.lastname: Pegington
authors.affiliation: Breast Cancer Now Research Unit, Institute of Cancer Sciences, Academic Health Science Centre, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
authors.forename: Katherine
authors.initials: K
authors.lastname: Spence
authors.affiliation: Breast Cancer Now Research Unit, Institute of Cancer Sciences, Academic Health Science Centre, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
authors.forename: Adam
authors.initials: A
authors.lastname: Mitchell
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Princess St, Manchester, M1 7DN, UK.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Vaughan
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Princess St, Manchester, M1 7DN, UK.
authors.forename: J William
authors.initials: JW
authors.lastname: Allwood
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Princess St, Manchester, M1 7DN, UK.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Princess St, Manchester, M1 7DN, UK.
authors.forename: Nicolas J W
authors.initials: NJ
authors.lastname: Rattray
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Princess St, Manchester, M1 7DN, UK.
authors.forename: Royston
authors.initials: R
authors.lastname: Goodacre
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK.|The Christie NHS Foundation Trust, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
authors.forename: D Gareth R
authors.initials: DG
authors.lastname: Evans
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK.
authors.forename: Ellen
authors.initials: E
authors.lastname: Mitchell
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK.
authors.forename: Debbie
authors.initials: D
authors.lastname: McMullen
authors.affiliation: Breast Cancer Now Research Unit, Institute of Cancer Sciences, Academic Health Science Centre, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
authors.forename: Robert B
authors.initials: RB
authors.lastname: Clarke
authors.affiliation: Genesis Breast Cancer Prevention Centre, University Hospital of South Manchester NHS Foundation Trust, Southmoor Road, Manchester, M23 9LT, UK.|Breast Cancer Now Research Unit, Institute of Cancer Sciences, Academic Health Science Centre, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.|The Christie NHS Foundation Trust, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
authors.forename: Anthony
authors.initials: A
authors.lastname: Howell
mesh_terms: D000328:Adult; D015415:Biomarkers; D001706:Biopsy; D001823:Body Composition; D001835:Body Weight; D001943:Breast Neoplasms / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D031204:Caloric Restriction; D016000:Cluster Analysis; D004734:Energy Metabolism; D005260:Female; D005786:Gene Expression Regulation; D006728:Hormones / Q000097:blood; D006801:Humans; D007333:Insulin Resistance; D008055:Lipids / Q000097:blood; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D042361:Mammary Glands, Human / Q000378:metabolism*; D008597:Menstrual Cycle; D055432:Metabolomics / Q000379:methods; D008875:Middle Aged; D019655:Quantitative Trait, Heritable
chemical_list: D015415:Biomarkers; D006728:Hormones; D008055:Lipids
keywords: Energy restriction; Gene expression; Metabolomics