title: Breast Cancer Cell Colonization of the Human Bone Marrow Adipose Tissue Niche.
abstract: OBJECTIVE
  Bone is a preferred site of breast cancer metastasis, suggesting the presence of tissue-specific features that attract and promote the outgrowth of breast cancer cells. We sought to identify parameters of human bone tissue associated with breast cancer cell osteotropism and colonization in the metastatic niche.
  
  
  METHODS
  Migration and colonization patterns of MDA-MB-231-fLuc-EGFP (luciferase-enhanced green fluorescence protein) and MCF-7-fLuc-EGFP breast cancer cells were studied in co-culture with cancellous bone tissue fragments isolated from 14 hip arthroplasties. Breast cancer cell migration into tissues and toward tissue-conditioned medium was measured in Transwell migration chambers using bioluminescence imaging and analyzed as a function of secreted factors measured by multiplex immunoassay. Patterns of breast cancer cell colonization were evaluated with fluorescence microscopy and immunohistochemistry.
  
  
  RESULTS
  Enhanced MDA-MB-231-fLuc-EGFP breast cancer cell migration to bone-conditioned versus control medium was observed in 12/14 specimens (P = .0014) and correlated significantly with increasing levels of the adipokines/cytokines leptin (P = .006) and IL-1β (P = .001) in univariate and multivariate regression analyses. Fluorescence microscopy and immunohistochemistry of fragments underscored the extreme adiposity of adult human bone tissues and revealed extensive breast cancer cell colonization within the marrow adipose tissue compartment.
  
  
  CONCLUSIONS
  Our results show that breast cancer cells migrate to human bone tissue-conditioned medium in association with increasing levels of leptin and IL-1β, and colonize the bone marrow adipose tissue compartment of cultured fragments. Bone marrow adipose tissue and its molecular signals may be important but understudied components of the breast cancer metastatic niche.
authors.affiliation: Department of Pediatrics, 150E Clark Center, 318 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5427. Electronic address: zstemple@stanford.edu.
authors.forename: Zach S
authors.initials: ZS
authors.lastname: Templeton
authors.affiliation: EMD Millipore Corporation, 14 Research Park Drive, St Charles, MO 63304-5618. Electronic address: Wen-rong.lie@emdmillipore.com.
authors.forename: Wen-Rong
authors.initials: WR
authors.lastname: Lie
authors.affiliation: Department of Immunology, Fairchild Science Building, D033, 299 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5124. Electronic address: Weiqi@stanford.edu.
authors.forename: Weiqi
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Immunology, Fairchild Science Building, D033, 299 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5124. Electronic address: Yaelhr@stanford.edu.
authors.forename: Yael
authors.initials: Y
authors.lastname: Rosenberg-Hasson
authors.affiliation: Department of Pediatrics, 150E Clark Center, 318 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5427. Electronic address: rvalluri@stanford.edu.
authors.forename: Rajiv V
authors.initials: RV
authors.lastname: Alluri
authors.affiliation: Department of Biomedical Data Science, Room T101F, Redwood Building, 150 Governor's Lane, Stanford University School of Medicine, Stanford, CA 94305-5405. Electronic address: John.s.tamaresis@stanford.edu.
authors.forename: John S
authors.initials: JS
authors.lastname: Tamaresis
authors.affiliation: Department of Pediatrics, 150E Clark Center, 318 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5427. Electronic address: mhb@stanford.edu.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Bachmann
authors.affiliation: Cell Sciences Imaging Facility, Beckman Center (B050B), Stanford University School of Medicine, Stanford, CA 94305-5301. Electronic address: microscopy@stanford.edu.
authors.forename: Kitty
authors.initials: K
authors.lastname: Lee
authors.affiliation: Department of Orthopaedic Surgery, 450 Broadway Street, Pavillion C, 4th Floor, Stanford University School of Medicine, Redwood City, CA 94063-6342. Electronic address: wmaloney@stanford.edu.
authors.forename: William J
authors.initials: WJ
authors.lastname: Maloney
authors.affiliation: Department of Pediatrics, 150E Clark Center, 318 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5427. Electronic address: ccontag@stanford.edu.
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
authors.affiliation: Department of Pediatrics, 150E Clark Center, 318 Campus Drive, Stanford University School of Medicine, Stanford, CA 94305-5427. Electronic address: bonnie.king@stanford.edu.
authors.forename: Bonnie L
authors.initials: BL
authors.lastname: King
mesh_terms: D017667:Adipocytes / Q000378:metabolism; D000273:Adipose Tissue / Q000378:metabolism / Q000473:pathology*; D001853:Bone Marrow / Q000378:metabolism / Q000473:pathology*; D001842:Bone and Bones / Q000378:metabolism / Q000473:pathology; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D060833:Cellular Microenvironment; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D053583:Interleukin-1beta / Q000378:metabolism; D020738:Leptin / Q000378:metabolism; D016015:Logistic Models; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D061986:MCF-7 Cells; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015999:Multivariate Analysis
chemical_list: D017077:Culture Media, Conditioned; D053583:Interleukin-1beta; D020738:Leptin; D049452:Green Fluorescent Proteins; D008156:Luciferases