A F Chambers

The University of Western Ontario, London, Ontario, Canada

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Publications (260)1180.1 Total impact

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    ABSTRACT: Tumor-derived matricellular proteins such as osteopontin (OPN) and tenascin-C (TN-C) have been implicated in tumor growth and metastasis. However, the molecular basis of how these proteins contribute to tumor progression remains to be elucidated. Importantly, these matricellular proteins are known to interact with α9β1 integrin. Therefore, we hypoth- esized that tumor-derived α9β1 integrin may contribute to tumor progression. To clarify the roles of α9β1 integrin in tumor growth and lymphatic metastasis, we used an inhibitory anti-human α9β1 integrin antibody (anti-hα9β1 antibody) and a α9β1 integrin-positive human breast cancer cell line, MDA-MB-231 luc-D3H2LN (D3H2LN), in vitro functional assays, and an in vivo orthotopic xenotransplantation model.
    Journal of Molecular Medicine 01/2015; · 4.77 Impact Factor
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    ABSTRACT: Planar cell polarity (PCP) signaling has been shown in different studies to either promote or inhibit the malignancy of breast cancer. Using the 21T cell lines, which were derived from an individual patient and represent distinct stages of progression, we show that the prototypical PCP ligand, WNT5A, is expressed highest in 21MT-1 cells (invasive mammary carcinoma) and lowest in 21PT (atypical ductal hyperplasia) and 21NT (ductal carcinoma in situ) cells. Overexpression of WNT5A decreased spherical colony formation and increased invasion and in vivo extravasation only in 21NT cells; whereas overexpression increased migration of both 21PT and 21NT cells. WNT5A overexpression also increased RHOA expression of both cell lines and subsequent RHOA knockdown blocked WNT5A-induced migration, but only partially blocked WNT5A-induced invasion of 21NT cells. PCP can signal through VANGL1 to modulate AP-1 target genes (e.g. MMP3) and induce invasion. VANGL1 knockdown inhibited WNT5A-induced invasion of 21NT cells, but had no effect on WNT5A-induced migration of either 21PT or 21NT cells. WNT5A-induced MMP3 expression was seen only in 21NT cells, an effect that was VANGL1 dependent, but independent of AP-1. We thus provide evidence that PCP signaling can act in a context dependent manner to promote breast cancer progression.
    Scientific Reports 09/2014; 4:6315. · 5.08 Impact Factor
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    ABSTRACT: Tumor cell extravasation is a key step during cancer metastasis, yet the precise mechanisms that regulate this dynamic process are unclear. We utilized a high-resolution time-lapse intravital imaging approach to visualize the dynamics of cancer cell extravasation in vivo. During intravascular migration, cancer cells form protrusive structures identified as invadopodia by their enrichment of MT1-MMP, cortactin, Tks4, and importantly Tks5, which localizes exclusively to invadopodia. Cancer cells extend invadopodia through the endothelium into the extravascular stroma prior to their extravasation at endothelial junctions. Genetic or pharmacological inhibition of invadopodia initiation (cortactin), maturation (Tks5), or function (Tks4) resulted in an abrogation of cancer cell extravasation and metastatic colony formation in an experimental mouse lung metastasis model. This provides direct evidence of a functional role for invadopodia during cancer cell extravasation and distant metastasis and reveals an opportunity for therapeutic intervention in this clinically important process.
    Cell Reports 08/2014; · 7.21 Impact Factor
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    ABSTRACT: Tumor cells have unstable genomes relative to non-tumor cells. Decreased DNA integrity resulting from tumor cell instability is important in generating favorable therapeutic indices, and intact DNA repair mediates resistance to therapy. Targeting DNA repair to promote the action of anti-cancer agents is therefore an attractive therapeutic strategy. BRCA2 is involved in homologous recombination repair. BRCA2 defects increase cancer risk but, paradoxically, cancer patients with BRCA2 mutations have better survival rates. We queried TCGA data and found that BRCA2 alterations led to increased survival in patients with ovarian and endometrial cancer. We developed a BRCA2-targeting second-generation antisense oligonucleotide (ASO), which sensitized human lung, ovarian, and breast cancer cells to cisplatin by as much as 60%. BRCA2 ASO treatment overcame acquired cisplatin resistance in head and neck cancer cells, but induced minimal cisplatin sensitivity in non-tumor cells. BRCA2 ASO plus cisplatin reduced respiration as an early event preceding cell death, concurrent with increased glucose uptake without a difference in glycolysis. BRCA2 ASO and cisplatin decreased metastatic frequency in vivo by 77%. These results implicate BRCA2 as a regulator of metastatic frequency and cellular metabolic response following cisplatin treatment. BRCA2 ASO, in combination with cisplatin, is a potential therapeutic anti-cancer agent.
    Molecular oncology. 06/2014;
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    ABSTRACT: Our previous work showed that Cyclin A2 deficiency promotes cell invasion in fibroblasts. Given that the majority of cancers emerge from epithelia, we explored novel functions for Cyclin A2 by depleting it in normal mammary epithelial cells. This caused an epithelial to mesenchymal transition (EMT) associated with loss of cell-to-cell contacts, decreased E-Cadherin expression and increased invasive properties characterized by a reciprocal regulation of RhoA and RhoC activities, where RhoA-decreased activity drove cell invasiveness and E-Cadherin delocalization, and RhoC-increased activity only supported cell motility. Phenotypes induced by Cyclin A2 deficiency were exacerbated upon oncogenic activated-Ras expression, which led to an increased expression of EMT-related transcriptional factors. Moreover, Cyclin A2-depleted cells exhibited stem cell-like properties and increased invasion in an in vivo avian embryo model. Our work supports a model where Cyclin A2 downregulation facilitates cancer cell EMT and metastatic dissemination.
    Cellular and Molecular Life Sciences CMLS 05/2014; · 5.62 Impact Factor
  • International Society of Magnetic Resonance in Medicine (ISMRM) 22nd Scientific Meeting and Exhibition, Milan, Italy; 05/2014
  • Acta oncologica (Stockholm, Sweden) 01/2014; · 2.27 Impact Factor
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    ABSTRACT: Osteopontin (OPN) is a malignancy-associated glycoprotein that contributes functionally to tumor aggressiveness. In metastatic breast cancer, we previously demonstrated that elevated OPN in primary tumor and blood was associated with poor prognosis. We measured OPN in plasma by ELISA, and in tumor by immunohistochemistry, in 624 (94%) and 462 (69%) respectively of 667 postmenopausal women with hormone responsive early breast cancer treated by surgery followed by adjuvant treatment with tamoxifen +/- octreotide in a randomized trial (NCIC CTG MA.14). Plasma OPN was measured in 2,540 samples; 688 at baseline and 1,852 collected during follow-up. Mean baseline plasma OPN was 46 ng/ml (range 22.6 - 290) which did not differ from normal levels. Mean percentage OPN tumor cell positivity was 33.9 (95% CI 30.2 - 37.9). There was no correlation between plasma and tumor OPN values. In multivariate analysis, neither was associated with event-free survival (EFS), relapse-free survival (RFS), overall survival (OS), bone RFS or non-bone RFS. An exploratory analysis in patients with recurrence showed higher mean OPN plasma levels 60.7 ng/ml (23.9 - 543) in the recurrence period compared with baseline levels. The hypothesis that OPN tumor expression would have independent prognostic value in early breast cancer was not supported by multivariate analysis of this study population. Plasma OPN levels in women with hormone responsive early breast cancer in the MA.14 trial were not elevated and there was no evidence for prognostic value of plasma OPN in this defined group of patients. However, our finding of elevated mean OPN plasma level around the time of recurrence warrants further study.Trial registration: NCT00002864.
    Breast cancer research: BCR 01/2014; 16(1):R8. · 5.87 Impact Factor
  • Hon S Leong, Ann F Chambers
    Proceedings of the National Academy of Sciences 01/2014; · 9.81 Impact Factor
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    ABSTRACT: Objectives Osteopontin (OPN) plays a role in tumor progression. This study aimed to determine the expression of OPN, CD44 and integrin αvβ3, in pleomorphic adenoma (PA), acinic cell carcinoma (ACA), and mucoepidermoid carcinoma (MEC). Study Design Immunohistochemistry was used to semi-quantify the level of OPN expression and its receptors in normal salivary glands (NSG; n = 20), PA (n = 20), ACA (n = 11) and MEC (n = 29). Results OPN expression was increased in ACA and MEC compared to PA and NSG (median scores 6, 6, 4 and 4 respectively). CD44 expression was increased in ACA, and reduced in MEC and PA compared with NSG (median scores 8, 4, 3 and 5 respectively) Integrin αvβ3 median scores were 5 in ACA, 1 in MEC and 0 in PA and NSG. Conclusions OPN is expressed in salivary gland tumors, and is at higher levels in ACA and MEC.
    Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology 01/2014; · 1.50 Impact Factor
  • Donna H Murrell, Paula J Foster, Ann F Chambers
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    ABSTRACT: Breast cancer that has metastasized to the brain presents difficult clinical challenges. This diagnosis comes with high mortality rates, largely due to complexities in early detection and ineffective therapies associated with both dormancy and impermeability of the blood-brain barrier (BBB). Magnetic resonance imaging (MRI) is the current gold standard for diagnosis and assessment of brain tumors. It has been used clinically to investigate metastatic development as well as monitor response to therapy. Here, we describe preclinical imaging strategies that we have used to study the development of brain metastases due to breast cancer. Using this approach, we have identified three subsets of metastatic disease: permeable metastases, nonpermeable metastases, and solitary, dormant cancer cells, which likely have very different biology and responses to therapy. The ability to simultaneously monitor the spatial and temporal distribution of dormant cancer cells, metastatic growth, and associated tumor permeability can provide great insight into factors that contribute to malignant proliferation. Our preclinical findings suggest that standard clinical detection strategies may underestimate the true metastatic burden of breast cancer that has metastasized to the brain. A better understanding of true metastatic burden in brains will be important to assist in the development of more effective chemotherapeutics-particularly those targeted to cross the BBB-as well as detection of small nonpermeable metastases.
    Journal of Molecular Medicine 12/2013; · 4.77 Impact Factor
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    ABSTRACT: Osteopontin (OPN) plays a role in tumor progression. This study aimed to determine the expression of OPN, CD44, and integrin αvβ3 in pleomorphic adenoma (PA), polymorphous low-grade adenocarcinoma (PLGA), and adenoid cystic carcinoma (ACC). Immunohistochemistry was used to semiquantify the level of expression of OPN and its receptors in normal salivary glands (NSG; n = 20), PA (n = 20), PLGA (n = 16), and ACC (n = 22). OPN expression was increased in PLGA and intermediate-/high-grade ACC compared with PA and NSG (median scores, 6, 5, 4, and 4, respectively). CD44 expression was reduced in PA, PLGA, and ACC. OPN expression levels were moderately correlated with CD44 in PLGA. Integrin αvβ3 was not expressed in PA and ACC and was seen in only 1 case of PLGA. OPN is expressed in salivary gland tumors but does not correlate well with CD44 and αvβ3.
    Oral surgery, oral medicine, oral pathology and oral radiology. 12/2013; 116(6):743-51.
  • World Molecular Imaging Congress, 2013, Savannah, USA; 09/2013
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    ABSTRACT: Flow quantification with high-frequency power Doppler ultrasound can be performed using the wall-filter selection curve (WFSC) method [Elfarnawany et al., Ultrasound Med. Biol. 38, 1429-1439 (2012)]. The WFSC method plots color pixel density (CPD) as a function of wall filter cut-off velocity as a means of objectively selecting an operating point cut-off velocity. In this study, an in vivo video microscopy (IVVM) system was used to measure the size of small (140-400 μm diameter) mouse testicular vessels immediately after the vessels were imaged with 30 MHz power Doppler. The mouse remained on the same platform throughout ultrasound and IVVM imaging. Measurements in four image planes from three mice demonstrated that, similar to previously reported flow-phantom data, in vivo WFSCs exhibit distinct, sloped "characteristic intervals" at cut-off velocities where the CPD approaches the gold-standard IVVM estimate of vascular volume fraction. A wide range of operating point cut-off velocities (4.5 to 12 mm/s) was obtained, which indicates that use of a predetermined cut-off can produce substantial errors in cross-sectional studies that employ power Doppler to quantify vascularity. The WFSC method is a promising strategy for adapting the cut-off velocity to intersubject and longitudinal variations in blood flow during microvascular imaging experiments.
    The Journal of the Acoustical Society of America 05/2013; 133(5):3540. · 1.65 Impact Factor
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    ABSTRACT: Contrast-enhanced ultrasound (CEUS) serves oncology by imaging tumor blood supply to enable quantification of longitudinal vascular changes and monitoring of treatment responses. Unfortunately, the linear subtraction methods commonly used for preclinical imaging are susceptible to registration errors and motion artifacts that lead to reduced contrast-to-tissue ratios. In this presentation, an alternative approach is proposed to improve discrimination between the contrast and tissue signals by comparing the first-order speckle statistics of images acquired before and after injection of microbubbles. The microbubble signal component is modeled as a temporally varying random process superimposed on a Rayleigh-distributed speckle signal representing backscatter from tissue. Images were acquired at 18 MHz from a murine orthotopic (mammary fat pad) xenograft breast cancer model following a bolus injection of microbubbles. Images were processed using gold-standard pulse inversion nonlinear CEUS, conventional linear subtraction, and the proposed statistical method. In comparison to conventional linear CEUS, the statistical method produced a wash-in curve that showed closer agreement to the gold-standard nonlinear CEUS data. The statistical method eliminates baseline image subtraction from linear CEUS processing, which should streamline the imaging workflow, improve the robustness of image quantification, and enable real-time perfusion imaging with linear CEUS.
    The Journal of the Acoustical Society of America 05/2013; 133(5):3263. · 1.65 Impact Factor
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    ABSTRACT: High plasma osteopontin (OPN) has been linked to tumour hypoxia, metastasis, and poor prognosis. This study aims to assess whether plasma osteopontin was a biomarker of increasing progression within prostate cancer (PCa) prognostic groups and whether it reflected treatment response to local and systemic therapies. Baseline OPN was determined in men with localised (n=199), locally recurrent (n=9) and castrate-resistant, metastatic PCa (CRPC-MET; n=37). Receiver-operating curves (ROC) were generated to describe the accuracy of OPN for distinguishing between localised risk groups or localised vs metastatic disease. We also measured OPN pre- and posttreatment, following radical prostatectomy, external beam radiotherapy (EBRT), androgen deprivation (AD) or taxane-based chemotherapy. The CRPC-MET patients had increased baseline values (mean 219; 56-513 ng ml(-1); P<0.0001) compared with the localised, non-metastatic group (mean 72; 12-438 ng ml(-1)). The area under the ROC to differentiate localised vs metastatic disease was improved when OPN was added to prostate-specific antigen (PSA) (0.943-0.969). Osteopontin neither distinguished high-risk PCa from other localised PCa nor correlated with serum PSA at baseline. Osteopontin levels reduced in low-risk patients after radical prostatectomy (P=0.005) and in CRPC-MET patients after chemotherapy (P=0.027), but not after EBRT or AD. Plasma OPN is as good as PSA at predicting treatment response in CRPC-MET patients after chemotherapy. Our data do not support the use of plasma OPN as a biomarker of increasing tumour burden within localised PCa.
    British Journal of Cancer 08/2012; 107(5):840-6. · 5.08 Impact Factor
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    ABSTRACT: The overall goal of this study was to assess the utility of three-dimensional magnetic resonance imaging (MRI) for monitoring the temporal and spatial development of experimental brain metastasis in mice. Brain metastatic human breast cancer cells (231-BR or 231-BR-HER2) were injected intracardially in nude mice for delivery to the brain. Mouse brains were imaged in vivo at different time points using a balanced steady-state-free precession (bSSFP) pulse sequence at 1.5 T. Brains were categorized into four regions: cortex, central brain, olfactory, and posterior. The number of metastases and their volumes were quantified for both cell lines. There was no difference in the mean number of metastases for either cell line. The volumes of metastases in mice injected with 231-BR-HER2 cells were significantly larger than those for mice injected with 231-BR cells. The growth rate for 231-BR-HER2 metastases was 67.5% compared with 54.4% for the 231-BR metastases. More than 50% of metastases were located in the cortex and 25% to 30% of metastases were identified in the central brain for each time point and for mice injected with either cell line. The volumes of metastases were significantly larger in mice with fewer metastases at end point. SIGNIFICANT CONCLUSIONS: MRI provided a comprehensive accounting of the number and size of experimental brain metastases in the whole mouse brain at multiple time points. This approach has provided new information about the temporal and spatial development of metastases in the brain not possible by other histopathologic or imaging methods.
    Translational oncology 06/2012; 5(3):217-25. · 3.40 Impact Factor
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    ABSTRACT: Lymphatic metastasis is a common occurrence in human breast cancer, mechanisms remaining poorly understood. MDA-MB-468LN (468LN), a variant of the MDA-MB-468GFP (468GFP) human breast cancer cell line, produces extensive lymphatic metastasis in nude mice. 468LN cells differentially express α9β1 integrin, a receptor for lymphangiogenic factors VEGF-C/-D. We explored whether (1) differential production of VEGF-C/-D by 468LN cells provides an autocrine stimulus for cellular motility by interacting with α9β1 and a paracrine stimulus for lymphangiogenesis in vitro as measured with capillary-like tube formation by human lymphatic endothelial cells (HMVEC-dLy); (2) differential expression of α9 also promotes cellular motility/invasiveness by interacting with macrophage derived factors; (3) stable knock-down of VEGF-D or α9 in 468LN cells abrogates lymphangiogenesis and lymphatic metastasis in vivo in nude mice. A comparison of expression of cyclo-oxygenase (COX)-2 (a VEGF-C/-D inducer), VEGF-C/-D and their receptors revealed little COX-2 expression by either cells. However, 468LN cells showed differential VEGF-D and α9β1 expression, VEGF-D secretion, proliferative, migratory/invasive capacities, latter functions being stimulated further with VEGF-D. The requirement of α9β1 for native and VEGF-D-stimulated proliferation, migration and Erk activation was demonstrated by treating with α9β1 blocking antibody or knock-down of α9. An autocrine role of VEGF-D in migration was shown by its impairment by silencing VEGF-D and restoration with VEGF-D. 468LN cells and their soluble products stimulated tube formation, migration/invasiveness of HMVEC-dLy cell in a VEGF-D dependent manner as indicated by the loss of stimulation by silencing VEGF-D in 468LN cells. Furthermore, 468LN cells showed α9-dependent stimulation of migration/invasiveness by macrophage products. Finally, capacity for intra-tumoral lymphangiogenesis and lymphatic metastasis in nude mice was completely abrogated by stable knock-down of either VEGF-D or α9 in 468LN cells. Differential capacity for VEGF-D production and α9β1 integrin expression by 468LN cells jointly contributed to their lymphatic metastatic phenotype.
    PLoS ONE 01/2012; 7(4):e35094. · 3.53 Impact Factor
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    ABSTRACT: The analysis of dynamic events in the tumor microenvironment during cancer progression is limited by the complexity of current in vivo imaging models. This is coupled with an inability to rapidly modulate and visualize protein activity in real time and to understand the consequence of these perturbations in vivo. We developed an intravital imaging approach that allows the rapid induction and subsequent depletion of target protein levels within human cancer xenografts while assessing the impact on cell behavior and morphology in real time. A conditionally stabilized fluorescent E-cadherin chimera was expressed in metastatic breast cancer cells, and the impact of E-cadherin induction and depletion was visualized using real-time confocal microscopy in a xenograft avian embryo model. We demonstrate the assessment of protein localization, cell morphology and migration in cells undergoing epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors. This technique allows for precise control over protein activity in vivo while permitting the temporal analysis of dynamic biophysical parameters.
    PLoS ONE 01/2012; 7(1):e30177. · 3.53 Impact Factor
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    Hon Sing Leong, Ann F Chambers, John D Lewis
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    ABSTRACT: Cell migration and metastasis are key features of aggressive tumors. These processes can be difficult to study, as they often occur deep within the body of a cancer patient or an experimental animal. In vitro assays are able to model some aspects of these processes, and a number of assays have been developed to assess cancer cell motility, migration, and invasion. However, in vitro assays have inherent limitations that may miss important aspects of these processes as they occur in vivo. The chick embryo provides a powerful model for studying these processes in vivo, facilitated by the external and accessible nature of the chorioallantoic membrane (CAM), a well-vascularized tissue that surrounds the embryo. When coupled with multiple fluorescent approaches to labeling both cancer cells and the embryonic vasculature, along with image analysis tools, the chick CAM model offers cost-effective, rapid assays for studying cancer cell migration and metastasis in a physiologically-relevant, in vivo setting. Here, we present recent developments of detailed procedures for using shell-less chick embryos, coupled with fluorescent labeling of cancer cells and/or chick vasculature, to study cancer cell migration and metastasis in vivo.
    Methods in molecular biology (Clifton, N.J.) 01/2012; 872:1-14. · 1.29 Impact Factor

Publication Stats

12k Citations
1,180.10 Total Impact Points

Institutions

  • 1985–2014
    • The University of Western Ontario
      • • Schulich School of Medicine and Dentistry
      • • Department of Medical Biophysics
      • • Department of Oncology
      • • Department of Microbiology and Immunology
      • • Department of Biochemistry
      London, Ontario, Canada
  • 2006–2012
    • Robarts Research Institute
      • Imaging Research Laboratories
      London, Ontario, Canada
    • Tom Baker Cancer Centre
      Calgary, Alberta, Canada
  • 1987–2012
    • Regional Integration Cancer Center
      Мендоса, Mendoza, Argentina
  • 2010–2011
    • University of Haifa
      • Faculty of Natural Sciences
      Haifa, Haifa District, Israel
    • University of Toronto
      Toronto, Ontario, Canada
    • Massachusetts General Hospital
      Boston, Massachusetts, United States
  • 1997–2011
    • London Health Sciences Centre
      • Department of Pathology
      London, Ontario, Canada
    • The University of Warwick
      Coventry, England, United Kingdom
  • 2008
    • Harvard Medical School
      • Department of Medicine
      Boston, Massachusetts, United States
  • 1994–2004
    • Rutgers, The State University of New Jersey
      • • Department of Genetics
      • • Federated Departments of Biological Sciences
      New Brunswick, NJ, United States
  • 2003
    • Biomedical Research Institute, Rockville
      Maryland, United States
    • Moffitt Cancer Center
      Tampa, Florida, United States
  • 2002–2003
    • University of South Florida
      • Department of Pathology and Cell Biology
      Tampa, FL, United States
  • 1996–2002
    • Hokkaido University
      • • Institute for Genetic Medicine
      • • Institute of Immunological Science
      Sapporo-shi, Hokkaido, Japan
    • Nigerian Institute for Oil Palm Research
      Benim, Edo, Nigeria
  • 1992–2001
    • McGill University
      • Department of Surgery
      Montréal, Quebec, Canada
  • 2000
    • University of Alabama at Birmingham
      Birmingham, Alabama, United States
  • 1998
    • Salk Institute
      • Molecular Neurobiology Laboratory
      La Jolla, CA, United States
  • 1995
    • Cancer Centre London - Parkside
      Londinium, England, United Kingdom