Proteolytic Pathways in Progression of Pre-Malignant Breast Disease

乳腺癌前病变进展中的蛋白水解途径

基本信息

批准号：
7666266
负责人：
RAYMOND R MATTINGLY
金额：
$ 31.42万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7666266
关键词：
Algorithms American Cancer Society Architecture Atypical hyperplasia B-Lymphocytes Biological Markers Biopsy Breast Breast Diseases Carcinoma Cathepsins Cathepsins B Cell Line Cell Surface Receptors Cells Characteristics Coculture Techniques Cysteine Cysteine Protease Diagnosis Drug Industry Dysplasia Engineering Exhibits Fibroblasts Genes Goals Human In Vitro Indolent Intervention Lead Lesion Libraries Malignant - descriptor Malignant Neoplasms Mammary Ductal Carcinoma Mammary Gland Parenchyma Mammary gland Matrix Metalloproteinases Mediating Modeling Monomeric GTP-Binding Proteins Morphology Myoepithelial Myoepithelial cell Noninfiltrating Intraductal Carcinoma Pathway interactions Peptide Hydrolases Phenotype Phosphotransferases Premalignant Process Proteolysis Proteomics Reagent Recurrence Role Sampling Screening procedure Signal Transduction Specimen Staging Stromelysin 1 Sum Testing Therapeutic Therapeutic Intervention Woman Work Xenograft procedure cathepsin F cathepsin V cell type human disease implantation in vivo infiltrating duct carcinoma inhibitor/antagonist macrophage malignant breast neoplasm malignant phenotype neoplastic cell p21-activated kinase 1 post gamma-globulins public health relevance research clinical testing serial analysis of gene expression stromelysin 3 treatment strategy tumor

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：乳房X线检查会导致许多妇女被诊断出患有导管癌的原位[DCIS]，但我们无法准确预测哪些病变会发生恶性进展，以使侵袭性导管癌[IDC]或有效阻止这种过渡。人类乳房活检的研究与肿瘤细胞，巨噬细胞以及肌上皮细胞/[myo]成纤维细胞中的巨噬细胞中的Cysteine caltepsins V/L2和B有关。异常信号转导（例如通过P21激活的激酶1 [PAK1]）可能有助于细胞周围蛋白水解增加。我们的工作假设是，从侵入前DCIS到侵袭性癌的过渡和某些DCIS病变的快速发展是通过DCIS细胞和与DCIS相关细胞中蛋白水解途径的改变而介导的，而PAK1则导致这些异常的蛋白质蛋白途径的诱导有助于诱导PAK1。为了检验这一假设，我们将使用体外和体内进展模型来概括从侵入前DCI到侵入性癌的过渡，这些模型已指定为乳腺体系结构微环境工程。在这些模型中，我们将使用ISEGENIC MCF10细胞系[AT1，DCIS1和CA1D]和两个人DCIS细胞系[SUM-102和SUM-225]。我们的具体目的是：1。通过直接靶向和通过在PAK1水平下进行干预，并确定使用体外模仿模型是否改变了侵入性表型。 2。确定使用体外妈妈模型是否可以通过与肌上皮细胞（MyO]成纤维细胞或两种细胞类型一起共培养AIM 1，使用野生型细胞以及使用野生型细胞以及伴半胱胺cantepeine cantepsin F，K或L的表达和活性来改变侵袭性表型； 3。确定使用体内乳mame模型是否可以通过同时植入肌上皮细胞（MyO]成纤维细胞或两种细胞类型，使用野生型细胞和野生型细胞和伴有野生型的表达和活性来改变AIM 1的异种移植物的恶性表型。和4。筛选[通过我们的HU/MU PROTIN芯片]蛋白水解途径的体内乳mame模型，可能有助于从DCIS到IDC的过渡，并使用体外MAME模型来定义来自蛋白水解途径中心试剂库的功能变化。在肿瘤及其微环境中验证蛋白酶是DCI向IDC发展的关键，以及调节它们的激酶途径，应确定治疗干预措施的潜在靶标，以及生物标志物，以区分将快速发展为IDC的DCIS病变。公共卫生相关性：蛋白酶和激酶是制药行业为制定包括癌症在内的人类疾病的新治疗策略的强化努力的主题。我们提出的研究将发现并验证在肿瘤微环境中活跃的蛋白酶途径，并介导过渡到成熟的恶性肿瘤和调节这些蛋白酶途径的激酶途径。我们预计，我们的研究将确定生物标志物以区分前病变，这些病变将发展为侵入性癌症并定义将消除该进展的靶标。