Genomes and Genes
Proteolytic Pathways in Progression of Pre-Malignant Breast Disease
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
- Functional expression of recombinant human stefin A in mammalian and bacterial cellsCatharine 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...
- Live-cell imaging of tumor proteolysis: impact of cellular and non-cellular microenvironmentJennifer 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...
- Inhibition of cathepsin B activity attenuates extracellular matrix degradation and inflammatory breast cancer invasionBernadette 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...
- Cathepsin B inhibition limits bone metastasis in breast cancerNimali 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...
- Chronic autophagy is a cellular adaptation to tumor acidic pH microenvironmentsJonathan 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...
- RNA-Seq of human breast ductal carcinoma in situ models reveals aldehyde dehydrogenase isoform 5A1 as a novel potential targetHitchintan 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...
- Acidity generated by the tumor microenvironment drives local invasionVeronica 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...
- Tyrosyl phosphorylated PAK1 regulates breast cancer cell motility in response to prolactin through filamin AAlan 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...
- Next-generation sequencing: a powerful tool for the discovery of molecular markers in breast ductal carcinoma in situHitchintan 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...
- Aborted autophagy and nonapoptotic death induced by farnesyl transferase inhibitor and lovastatinJonathan 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...
- Capturing and characterizing immune cells from breast tumor microenvironment: an innovative surgical approachMohamed El-Shinawi
Department of General Surgery, Faculty of Medicine, Ain Shams University, Cairo, Egypt
Ann Surg Oncol 17:2677-84. 2010....
- 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 cellsQuanwen Li
Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA
Neoplasia 10:1444-58. 2008....
- Imaging and quantifying the dynamics of tumor-associated proteolysisMansoureh 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...
- Functional live-cell imaging demonstrates that beta1-integrin promotes type IV collagen degradation by breast and prostate cancer cellsMansoureh 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...
- Disease progression and solid tumor survival: a transcriptome decoherence modelAdrian 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....
- Fibroblast hepatocyte growth factor promotes invasion of human mammary ductal carcinoma in situChristopher Jedeszko
Department of Pharmacology, Wayne State University, Detroit, Michigan, USA
Cancer Res 69:9148-55. 2009....
- Three-dimensional overlay culture models of human breast cancer reveal a critical sensitivity to mitogen-activated protein kinase kinase inhibitorsQuanwen Li
Wayne State University, Department of Pharmacology, 540 East Canfield Avenue, Detroit, MI 48201, USA
J Pharmacol Exp Ther 332:821-8. 2010....
- Chiral porphyrazine near-IR optical imaging agent exhibiting preferential tumor accumulationEvan 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...
- A novel geranylgeranyl transferase inhibitor in combination with lovastatin inhibits proliferation and induces autophagy in STS-26T MPNST cellsKomal M Sane
Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA
J Pharmacol Exp Ther 333:23-33. 2010....
- Interleukin-6 increases expression and secretion of cathepsin B by breast tumor-associated monocytesMona 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...
- Acid-mediated tumor proteolysis: contribution of cysteine cathepsinsJennifer 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. ..
- The Shelf Live Evaluation of Investigational Dosage FormsJonathan 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. ..
- Advancing Systems Approaches to Personal and Population Breast Cancer ScreeningAnna N A Tosteson; Fiscal Year: 2013..abstract_text> ..
- Myoepithelial cell differentiation defects in ductal carcinoma in situ (DCIS)Kornelia Polyak; Fiscal Year: 2013..abstract_text> ..
- Molecular Pathogenesis of Basal-like Breast CancerRichard J Baer; Fiscal Year: 2013..e., metastasis, and develop strategies for therapy. ..
- CELLULAR AND MOLECULAR MECHANISMS OF GASTROINTESTINAL CANCERSLopa 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. ..
- ROLES AND REGULATION OF P53Carol 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. ..
- Growth Control in Multiple MyelomaBart Barlogie; Fiscal Year: 2013..This work will be accomplished with access to 5 shared resource cores. ..
- University of Maryland Greenebaum Cancer Center Support GrantKevin 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. ..
- Mayo Clinic Breast Cancer SPOREJAMES NEWELL INGLE; Fiscal Year: 2013..abstract_text> ..
- Molecular Response and Imaging-based Combination Strategies for Optimal PDTTayyaba 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. ..
- Signaling in Inflammation, Stress, and TumorigenesisGEORGE ROBERT STARK; Fiscal Year: 2013..abstract_text> ..
- NUCLEAR STRUCTURE AND GENE EXPRESSIONJanet L Stein; Fiscal Year: 2013..abstract_text> ..
- INTEGRATIVE PATHOPHYSIOLOGY OF SOLID TUMORSRakesh K Jain; Fiscal Year: 2013..abstract_text> ..
- Mechanistic Pharmacology of Anti-Mitotics and Apoptosis RegulationTimothy 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. ..
- UNMC EPPLEY CANCER CENTER SUPPORT GRANTKenneth H Cowan; Fiscal Year: 2013....
- Model-based predictions of responses RTK Pathway therapiesJoe W Gray; Fiscal Year: 2013..abstract_text> ..
- The Role of Immune Cells in Breast Development and CancerSung Jin Huh; Fiscal Year: 2013....