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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光检查导致许多女性被诊断患有导管原位癌[DCIS]，但我们无法准确预测哪些病变将恶性进展为浸润性导管癌[IDC]或有效阻止这种转变。对人类乳腺活检的研究表明，肿瘤细胞和巨噬细胞中的半胱氨酸组织蛋白酶 V/L2 和 B 以及肌上皮细胞/[肌]成纤维细胞中的组织蛋白酶 F、K 和 L 参与了这一过程。异常信号转导，例如通过 p21 激活激酶 1 [PAK1]，可能会导致细胞周蛋白水解作用增加。我们的工作假设是，从浸润前 DCIS 到浸润性癌的转变以及一些 DCIS 病变的快速进展是通过 DCIS 细胞和 DCIS 相关细胞中蛋白水解途径的改变介导的，并且 PAK1 失调有助于诱导这些异常的蛋白水解途径。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浸润前 DCIS 到浸润性癌的转变，我们已将这些模型指定为用于乳腺结构微环境工程的 MAME。在这些模型中，我们将使用同基因 MCF10 细胞系 [AT1、DCIS1 和 CA1d] 和两种人类 DCIS 细胞系 [SUM-102 和 SUM-225]。我们的具体目标是： 1. 通过直接靶向和通过 PAK1 水平的干预来调节等基因和 SUM DCIS 细胞系中半胱氨酸组织蛋白酶 V 或 B 的表达和活性，并使用体外 MAME 模型确定侵袭表型是否改变； 2. 使用体外 MAME 模型确定是否可以通过将来自 Aim 1 的修饰细胞与肌上皮细胞、[肌]成纤维细胞或这两种细胞类型共培养来改变侵袭表型，使用野生型细胞和半胱氨酸组织蛋白酶 F、K 或 L 的表达和活性已被调节的细胞； 3.使用体内MAME模型确定来自Aim 1的修饰细胞的异种移植物的恶性表型是否可以通过使用野生型细胞和半胱氨酸组织蛋白酶F、K或L的表达和活性已被调节的细胞同时植入肌上皮细胞、[肌]成纤维细胞或两种细胞类型来改变； 4. [通过我们的 Hu/Mu ProtIn 芯片] 筛选可能有助于从 DCIS 向 IDC 转变的蛋白水解途径的体内 MAME 模型，并使用体外 MAME 模型通过蛋白水解途径中心的试剂库来定义功能变化。在肿瘤及其微环境的背景下验证 DCIS 进展为 IDC 的关键蛋白酶以及调节它们的激酶途径，应确定治疗干预的潜在靶标以及区分将迅速进展为 IDC 的 DCIS 病变的生物标志物。公共健康相关性：蛋白酶和激酶是制药行业为开发人类疾病（包括癌症）新治疗策略而大力努力的主题。我们提出的研究将发现并验证在肿瘤微环境中活跃并介导向成熟恶性肿瘤转变的蛋白酶途径，以及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来区分将进展为侵袭性癌症的癌前病变，并确定消除这种进展的目标。