Proteolytic Pathways in Progression of Pre-Malignant Breast Disease

Summary

Principal Investigator: Bonnie F Sloane
Abstract: Mammographic screening leads to many women being diagnosed with ductal carcinoma in situ [DCIS], yet we cannot accurately predict which lesions will undergo malignant progression to invasive ductal carcinomas [IDC] or effectively block this transition. Studies of human breast biopsies have implicated in this process cysteine cathepsins V/L2 and B in tumor cells and macrophages and cathepsins F, K and L in myoepithelial cells/[myo]fibroblasts. Aberrant signal transduction, for example through p21-activated kinase 1 [PAK1], may contribute to increased pericellular proteolysis. Our working hypothesis is that the transition from pre-invasive DCIS to invasive carcinomas and the rapid progression of some DCIS lesions are mediated through alterations in proteolytic pathways in DCIS cells and DCIS-associated cells, and that dysregulated PAK1 contributes to the induction of these aberrant proteolytic pathways. To test this hypothesis, we will recapitulate the transition from pre-invasive DCIS to invasive carcinoma using in vitro and in vivo progression models that we have designated MAME for mammary architecture microenvironment engineering. In these models, we will use isogenic MCF10 cell lines [AT1, DCIS1 and CA1d] and two human DCIS cell lines [SUM-102 and SUM-225]. Our specific aims are to: 1. Modulate expression and activity of cysteine cathepsin V or B in the isogenic and SUM DCIS cell lines, both by direct targeting and through intervention at the level of PAK1, and determine using the in vitro MAME model whether the invasive phenotype is altered;2. Determine using the in vitro MAME model whether the invasive phenotype can be altered by co-culturing modified cells from Aim 1 with myoepithelial cells, [myo]fibroblasts or both cell types, using wild-type cells and ones in which expression and activity of cysteine cathepsin F, K or L have been modulated;3. Determine using the in vivo MAME model whether the malignant phenotype of xenografts of modified cells from Aim 1 can be altered by simultaneous implantation of myoepithelial cells, [myo]fibroblasts or both cell types, using wild-type cells and ones in which expression and activity of cysteine cathepsin F, K or L have been modulated;and 4. Screen [via our Hu/Mu ProtIn chip] the in vivo MAME model for proteolytic pathways that may contribute to the transition from DCIS to IDC and use the in vitro MAME model to define functional changes with libraries of reagents from the Center on Proteolytic Pathways. Validating, in the context both of the tumor and its microenvironment, proteases key to progression of DCIS to IDC, and kinase pathways that regulate them, should identify potential targets for therapeutic intervention as well as biomarkers to distinguish DCIS lesions that will rapidly progress to IDC. PUBLIC HEALTH RELEVANCE: Proteases and kinases are the subject of intensive efforts by the pharmaceutical industry to develop new treatment strategies for human diseases, including cancer. Our proposed studies will discover and validate protease pathways that are active in the tumor microenvironment and that mediate the transition to a full-blown malignancy, and kinase pathways that regulate these protease pathways. We anticipate that our studies will identify biomarkers to distinguish premalignant lesions that will progress to invasive cancers and define targets that will abrogate that progression.
Funding Period: 2008-08-01 - 2014-05-31
more information: NIH RePORT

Top Publications

  1. pmc Functional expression of recombinant human stefin A in mammalian and bacterial cells
    Catharine C Calkins
    Department of Pharmacology, Wayne State University School of Medicine, 540 East Canfield, Detroit, MI 48201, USA
    Protein Expr Purif 52:463-71. 2007
  2. pmc Live-cell imaging of tumor proteolysis: impact of cellular and non-cellular microenvironment
    Jennifer M Rothberg
    Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA
    Biochim Biophys Acta 1824:123-32. 2012
  3. pmc Inhibition of cathepsin B activity attenuates extracellular matrix degradation and inflammatory breast cancer invasion
    Bernadette C Victor
    Department of Pharmacology, Wayne State University, Detroit, Michigan 48201, USA
    Breast Cancer Res 13:R115. 2011
  4. pmc Cathepsin B inhibition limits bone metastasis in breast cancer
    Nimali P Withana
    Research Division, Peter MacCallum Cancer Centre, East Melbourne, Australia
    Cancer Res 72:1199-209. 2012
  5. pmc Chronic autophagy is a cellular adaptation to tumor acidic pH microenvironments
    Jonathan W Wojtkowiak
    Department of Cancer Imaging and Metabolism, Analytic Microscopy Core Facility, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, Florida 33612, USA
    Cancer Res 72:3938-47. 2012
  6. pmc RNA-Seq of human breast ductal carcinoma in situ models reveals aldehyde dehydrogenase isoform 5A1 as a novel potential target
    Hitchintan Kaur
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan, USA
    PLoS ONE 7:e50249. 2012
  7. pmc Acidity generated by the tumor microenvironment drives local invasion
    Veronica Estrella
    Departments of Cancer Imaging and Metabolism, Radiology, and Analytic Microscopy Laboratory, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA
    Cancer Res 73:1524-35. 2013
  8. pmc Tyrosyl phosphorylated PAK1 regulates breast cancer cell motility in response to prolactin through filamin A
    Alan Hammer
    Department of Biological Sciences, University of Toledo, Toledo, OH 43606 3390, USA
    Mol Endocrinol 27:455-65. 2013
  9. pmc Next-generation sequencing: a powerful tool for the discovery of molecular markers in breast ductal carcinoma in situ
    Hitchintan Kaur
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201, USA
    Expert Rev Mol Diagn 13:151-65. 2013
  10. pmc Aborted autophagy and nonapoptotic death induced by farnesyl transferase inhibitor and lovastatin
    Jonathan W Wojtkowiak
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA
    J Pharmacol Exp Ther 337:65-74. 2011

Detail Information

Publications22

  1. pmc Functional expression of recombinant human stefin A in mammalian and bacterial cells
    Catharine C Calkins
    Department of Pharmacology, Wayne State University School of Medicine, 540 East Canfield, Detroit, MI 48201, USA
    Protein Expr Purif 52:463-71. 2007
    ..Expression in the mammalian cells, on the other hand, can provide a significant research tool to study the functional roles of stefin A in mammalian systems such as regulation of cysteine proteases...
  2. pmc Live-cell imaging of tumor proteolysis: impact of cellular and non-cellular microenvironment
    Jennifer M Rothberg
    Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA
    Biochim Biophys Acta 1824:123-32. 2012
    ..Here we will discuss the background that led to our present studies as well as the techniques and models that we employ. This article is part of a Special Issue entitled: Proteolysis 50 years after the discovery of lysosome...
  3. pmc Inhibition of cathepsin B activity attenuates extracellular matrix degradation and inflammatory breast cancer invasion
    Bernadette C Victor
    Department of Pharmacology, Wayne State University, Detroit, Michigan 48201, USA
    Breast Cancer Res 13:R115. 2011
    ..Breast cancer invasion has been linked to proteolytic activity at the tumor cell surface. Here we explored a role for active cathepsin B on the cell surface in the invasiveness of IBC...
  4. pmc Cathepsin B inhibition limits bone metastasis in breast cancer
    Nimali P Withana
    Research Division, Peter MacCallum Cancer Centre, East Melbourne, Australia
    Cancer Res 72:1199-209. 2012
    ..Together, our findings established a prometastatic role for cathepsin B in distant metastasis and illustrated the therapeutic benefits of its selective inhibition in vivo...
  5. pmc Chronic autophagy is a cellular adaptation to tumor acidic pH microenvironments
    Jonathan W Wojtkowiak
    Department of Cancer Imaging and Metabolism, Analytic Microscopy Core Facility, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, Florida 33612, USA
    Cancer Res 72:3938-47. 2012
    ..Taken together, these results argue that acidic conditions in the tumor microenvironment promote autophagy, and that chronic autophagy occurs as a survival adaptation in this setting...
  6. pmc RNA-Seq of human breast ductal carcinoma in situ models reveals aldehyde dehydrogenase isoform 5A1 as a novel potential target
    Hitchintan Kaur
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan, USA
    PLoS ONE 7:e50249. 2012
    ..These results suggest that ALDH5A1 may play an important role in DCIS and potentially serve as a novel molecular therapeutic target...
  7. pmc Acidity generated by the tumor microenvironment drives local invasion
    Veronica Estrella
    Departments of Cancer Imaging and Metabolism, Radiology, and Analytic Microscopy Laboratory, H Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA
    Cancer Res 73:1524-35. 2013
    ..Cancer Res; 73(5); 1524-35. ©2012 AACR...
  8. pmc Tyrosyl phosphorylated PAK1 regulates breast cancer cell motility in response to prolactin through filamin A
    Alan Hammer
    Department of Biological Sciences, University of Toledo, Toledo, OH 43606 3390, USA
    Mol Endocrinol 27:455-65. 2013
    ..Down-regulation of PAK1 or filamin A abolishes the effect of PRL on cell migration. Thus, our data presented here bring some insight into the mechanism of PRL-stimulated motility of breast cancer cells...
  9. pmc Next-generation sequencing: a powerful tool for the discovery of molecular markers in breast ductal carcinoma in situ
    Hitchintan Kaur
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201, USA
    Expert Rev Mol Diagn 13:151-65. 2013
    ..Here, the authors review the applications and challenges of NGS in discovering novel potential therapeutic targets and candidate biomarkers in the premalignant progression of breast cancer...
  10. pmc Aborted autophagy and nonapoptotic death induced by farnesyl transferase inhibitor and lovastatin
    Jonathan W Wojtkowiak
    Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA
    J Pharmacol Exp Ther 337:65-74. 2011
    ..These studies indicate that cotreatment of STS-26T cells with lovastatin and FTI-1 induces an abortive autophagic program and nonapoptotic cell death...
  11. pmc Capturing and characterizing immune cells from breast tumor microenvironment: an innovative surgical approach
    Mohamed El-Shinawi
    Department of General Surgery, Faculty of Medicine, Ain Shams University, Cairo, Egypt
    Ann Surg Oncol 17:2677-84. 2010
    ....
  12. pmc Restoration of E-cadherin cell-cell junctions requires both expression of E-cadherin and suppression of ERK MAP kinase activation in Ras-transformed breast epithelial cells
    Quanwen Li
    Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA
    Neoplasia 10:1444-58. 2008
    ....
  13. pmc Imaging and quantifying the dynamics of tumor-associated proteolysis
    Mansoureh Sameni
    Department of Pharmacology, School of Medicine, Wayne State University, 540 E Canfield, Detroit, MI 48201, USA
    Clin Exp Metastasis 26:299-309. 2009
    ..The assays and models described here could serve as screening platforms for the identification of proteolytic pathways that are potential therapeutic targets and for further development of technologies and imaging probes for in vivo use...
  14. pmc Functional live-cell imaging demonstrates that beta1-integrin promotes type IV collagen degradation by breast and prostate cancer cells
    Mansoureh Sameni
    Department of Pharmacology and Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA
    Mol Imaging 7:199-213. 2008
    ..This is the first study to demonstrate through functional live-cell imaging that downregulation of beta1-integrin expression and function reduces proteolysis of collagen IV by breast and prostate cancer cells...
  15. pmc Disease progression and solid tumor survival: a transcriptome decoherence model
    Adrian E Platts
    The Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI 48201, USA
    Mol Cell Probes 24:53-60. 2010
    ....
  16. pmc Fibroblast hepatocyte growth factor promotes invasion of human mammary ductal carcinoma in situ
    Christopher Jedeszko
    Department of Pharmacology, Wayne State University, Detroit, Michigan, USA
    Cancer Res 69:9148-55. 2009
    ....
  17. pmc Three-dimensional overlay culture models of human breast cancer reveal a critical sensitivity to mitogen-activated protein kinase kinase inhibitors
    Quanwen Li
    Wayne State University, Department of Pharmacology, 540 East Canfield Avenue, Detroit, MI 48201, USA
    J Pharmacol Exp Ther 332:821-8. 2010
    ....
  18. pmc Chiral porphyrazine near-IR optical imaging agent exhibiting preferential tumor accumulation
    Evan R Trivedi
    Departments of Chemistry, Northwestern University, Evanston, IL 60208, USA
    Proc Natl Acad Sci U S A 107:1284-8. 2010
    ..As the need for cholesterol, and thus LDL, is elevated in highly proliferative tumor cells over nontumorigenic cells, 247 has potential application for all such tumors...
  19. pmc A novel geranylgeranyl transferase inhibitor in combination with lovastatin inhibits proliferation and induces autophagy in STS-26T MPNST cells
    Komal M Sane
    Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA
    J Pharmacol Exp Ther 333:23-33. 2010
    ....
  20. pmc Interleukin-6 increases expression and secretion of cathepsin B by breast tumor-associated monocytes
    Mona M Mohamed
    Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt
    Cell Physiol Biochem 25:315-24. 2010
    ..Our data suggest a role for IL-6 in increased monocyte expression and secretion of CTSB in response to soluble factors secreted by breast cancer cells...
  21. pmc Acid-mediated tumor proteolysis: contribution of cysteine cathepsins
    Jennifer M Rothberg
    Cancer Biology Program, Wayne State University, Detroit, MI Department of Pharmacology, Wayne State University, Detroit, MI
    Neoplasia 15:1125-37. 2013
    ..Our results are consistent with the acid-mediated invasion hypothesis and with a role for cathepsin B in promoting degradation of a basement membrane protein substrate, i.e., type IV collagen, in an acidic peritumoral environment. ..

Research Grants30

  1. The Shelf Live Evaluation of Investigational Dosage Forms
    Jonathan White; Fiscal Year: 2013
    ..This contract is essential for continued assurance of the quality of drugs undergoing clinical investigation for different types of cancer by Cancer Therapeutics Evaluation Program. ..
  2. Advancing Systems Approaches to Personal and Population Breast Cancer Screening
    Anna N A Tosteson; Fiscal Year: 2013
    ..abstract_text> ..
  3. Myoepithelial cell differentiation defects in ductal carcinoma in situ (DCIS)
    Kornelia Polyak; Fiscal Year: 2013
    ..abstract_text> ..
  4. Molecular Pathogenesis of Basal-like Breast Cancer
    Richard J Baer; Fiscal Year: 2013
    ..e., metastasis, and develop strategies for therapy. ..
  5. CELLULAR AND MOLECULAR MECHANISMS OF GASTROINTESTINAL CANCERS
    Lopa Mishra; Fiscal Year: 2013
    ..Significantly, this program promises to yield new therapies targeted at these difficult-to-treat lethal cancers at the bench, and then to translate the results rapidly into clinical care, at the bedside. ..
  6. ROLES AND REGULATION OF P53
    Carol Prives; Fiscal Year: 2013
    ..The projects are even more interdependent and interactive than before and as a result much of the proposed research cannot be done effectively without the support of this program. ..
  7. Growth Control in Multiple Myeloma
    Bart Barlogie; Fiscal Year: 2013
    ..This work will be accomplished with access to 5 shared resource cores. ..
  8. University of Maryland Greenebaum Cancer Center Support Grant
    Kevin J Cullen; Fiscal Year: 2013
    ..Reflecting our remarkable and continued growth, UMGCC seeks to renew its CCSG to enhance and expand its efforts in high-quality and clinically relevant cancer research. ..
  9. Mayo Clinic Breast Cancer SPORE
    JAMES NEWELL INGLE; Fiscal Year: 2013
    ..abstract_text> ..
  10. Molecular Response and Imaging-based Combination Strategies for Optimal PDT
    Tayyaba Hasan; Fiscal Year: 2013
    ..NMSC on the other hand has many options but the high incidence puts a heavy burden on society in terms of cost and suffering. ..
  11. Signaling in Inflammation, Stress, and Tumorigenesis
    GEORGE ROBERT STARK; Fiscal Year: 2013
    ..abstract_text> ..
  12. NUCLEAR STRUCTURE AND GENE EXPRESSION
    Janet L Stein; Fiscal Year: 2013
    ..abstract_text> ..
  13. INTEGRATIVE PATHOPHYSIOLOGY OF SOLID TUMORS
    Rakesh K Jain; Fiscal Year: 2013
    ..abstract_text> ..
  14. Mechanistic Pharmacology of Anti-Mitotics and Apoptosis Regulation
    Timothy J Mitchison; Fiscal Year: 2013
    ..In aim 4 we will pursue several approaches towards translating mechanistic understanding from aims 1-3 into improved patient care. ..
  15. UNMC EPPLEY CANCER CENTER SUPPORT GRANT
    Kenneth H Cowan; Fiscal Year: 2013
    ....
  16. Model-based predictions of responses RTK Pathway therapies
    Joe W Gray; Fiscal Year: 2013
    ..abstract_text> ..
  17. The Role of Immune Cells in Breast Development and Cancer
    Sung Jin Huh; Fiscal Year: 2013
    ....