权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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：
8244681
负责人：
RAYMOND R MATTINGLY
金额：
$ 5.16万
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/8244681
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 Situ Lesion 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 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(申请人提供)：乳房X光检查导致许多女性被诊断为导管原位癌[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]或有效阻止这一转变。对人类乳腺活检的研究表明，半胱氨酸组织蛋白V/L2和B在肿瘤细胞和巨噬细胞中存在，组织蛋白F、K和L在肌上皮细胞/成纤维细胞中存在。异常的信号转导，例如通过p21激活的激酶1[PAK1]，可能有助于细胞周围蛋白分解的增加。我们的工作假设是，从DCIS前期到浸润性癌的转变以及某些DCIS病变的快速进展是通过DCIS细胞和DCIS相关细胞中蛋白分解途径的改变来调节的，而调控异常的PAK1参与了这些异常蛋白分解途径的诱导。为了验证这一假说，我们将使用我们指定的乳腺微环境工程MAME体外和体内进展模型来概括从浸润性癌前期到浸润性癌的转变。在这些模型中，我们将使用等基因的MCF10细胞系[AT1，DCIS1和CA1d]和两个人类DCIS细胞系[SUM-102和SUM-225]。我们的具体目的是：1.通过直接靶向和在PAK1水平上的干预，调节半胱氨酸组织蛋白酶V或B在等基因和全基因DCIS细胞系中的表达和活性，并利用体外MAME模型确定是否可以改变侵袭性表型；2.利用体外MAME模型确定是否可以将修饰后的AIM 1细胞与肌上皮细胞、成纤维细胞或两者共同培养，用野生型细胞和半胱氨酸组织蛋白酶F、K或L的表达和活性受到调控的细胞来改变侵袭性表型；3.使用体内MAME模型，利用野生型细胞和半胱氨酸组织蛋白F、K或L表达和活性已被调控的细胞，确定是否可以通过同时移植肌上皮细胞、成纤维细胞或两种细胞类型来改变AIM 1修饰细胞的异种移植的恶性表型；以及4.[通过我们的Hu/Mu Protin芯片]筛选可能有助于从DCIS向IDC转变的体内MAME模型，并使用体外MAME模型与Proteolytic Path Center的试剂库一起确定功能变化。在肿瘤及其微环境的背景下，验证DCIS向IDC进展的关键的蛋白水解酶，以及调节它们的激酶途径，应该确定潜在的治疗干预靶点以及生物标记物，以区分将迅速进展到IDC的DCIS病变。与公共卫生相关：蛋白酶和激酶是制药业为开发包括癌症在内的人类疾病的新治疗策略而进行的密集努力的主题。我们拟议的研究将发现并验证在肿瘤微环境中活跃的、介导向全面恶性转化的蛋白水解酶途径，以及调节这些蛋白水解酶途径的激酶途径。我们预计，我们的研究将识别生物标记物，以区分将进展为浸润性癌症的癌前病变，并确定将消除这种进展的靶点。