title: Cellular heterogeneity profiling by hyaluronan probes reveals an invasive but slow-growing breast tumor subset.
abstract: Tumor heterogeneity confounds cancer diagnosis and the outcome of therapy, necessitating analysis of tumor cell subsets within the tumor mass. Elevated expression of hyaluronan (HA) and HA receptors, receptor for HA-mediated motility (RHAMM)/HA-mediated motility receptor and cluster designation 44 (CD44), in breast tumors correlates with poor outcome. We hypothesized that a probe for detecting HA-HA receptor interactions may reveal breast cancer (BCa) cell heterogeneity relevant to tumor progression. A fluorescent HA (F-HA) probe containing a mixture of polymer sizes typical of tumor microenvironments (10-480 kDa), multiplexed profiling, and flow cytometry were used to monitor HA binding to BCa cell lines of different molecular subtypes. Formulae were developed to quantify binding heterogeneity and to measure invasion in vivo. Two subsets exhibiting differential binding (HA(-/low) vs. HA(high)) were isolated and characterized for morphology, growth, and invasion in culture and as xenografts in vivo. F-HA-binding amounts and degree of heterogeneity varied with BCa subtype, were highest in the malignant basal-like cell lines, and decreased upon reversion to a nonmalignant phenotype. Binding amounts correlated with CD44 and RHAMM displayed but binding heterogeneity appeared to arise from a differential ability of HA receptor-positive subpopulations to interact with F-HA. HA(high) subpopulations exhibited significantly higher local invasion and lung micrometastases but, unexpectedly, lower proliferation than either unsorted parental cells or the HA(-/low) subpopulation. Querying F-HA binding to aggressive tumor cells reveals a previously undetected form of heterogeneity that predicts invasive/metastatic behavior and that may aid both early identification of cancer patients susceptible to metastasis, and detection/therapy of invasive BCa subpopulations.
authors.affiliation: Life Sciences Division, E. O. Lawrence Berkeley National Laboratories, Berkeley, CA 94720.
authors.forename: Mandana
authors.initials: M
authors.lastname: Veiseh
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Kwon
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Borowsky
authors.forename: Cornelia
authors.initials: C
authors.lastname: Tolg
authors.forename: Hon S
authors.initials: HS
authors.lastname: Leong
authors.forename: John D
authors.initials: JD
authors.lastname: Lewis
authors.forename: Eva A
authors.initials: EA
authors.lastname: Turley
authors.forename: Mina J
authors.initials: MJ
authors.lastname: Bissell
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000145:classification* / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000378:metabolism; D003199:Computer Systems; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005260:Female; D005453:Fluorescence; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid; D051379:Mice; D016513:Mice, SCID; D015335:Molecular Probes; D009361:Neoplasm Invasiveness; D010641:Phenotype; D064726:Triple Negative Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D023041:Xenograft Model Antitumor Assays
chemical_list: D016326:Extracellular Matrix Proteins; D018960:Hyaluronan Receptors; D015335:Molecular Probes; C098946:hyaluronan-mediated motility receptor; D006820:Hyaluronic Acid
keywords: heterogeneity index; hyaluronan binding; tumor cell heterogeneity
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