Progression of DCIS to invasive breast cancer through CCR2 chemokine signaling

通过 CCR2 趋化因子信号传导将 DCIS 进展为浸润性乳腺癌

• 批准号: 9297227
• 负责人: FARIBA BEHBOD
• 金额: $ 41.35万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9297227
• 关键词: Affect; Apoptotic; Biochemical; Biology; Breast; Breast Cancer cell line; Breast Carcinoma; CC chemokine receptor 2; CCL2 gene; Carcinoma; Cell Culture System; Cell Line; Cell Survival; Cells; Development; Disease Progression; Ductal Carcinoma; Fibroblasts; Goals; Human; Injection of therapeutic agent; Lead; Lesion; Ligand Binding; MAP Kinase Gene; Malignant Epithelial Cell; Mammary Neoplasms; Mammary gland; Mediating; Modeling; Molecular; Mus; Non-Invasive Lesion; Noninfiltrating Intraductal Carcinoma; Normal tissue morphology; Pathway interactions; Patients; Phenotype; Proteins; Receptor Signaling; Recruitment Activity; Risk; Role; Sampling; Signal Transduction; Therapeutic; Transplantation; Woman; Xenograft Model; autocrine; base; breast cancer diagnosis; capsule; cell motility; chemokine; chemokine receptor; in vivo; infiltrating duct carcinoma; insight; knock-down; macrophage; malignant breast neoplasm; mouse model; novel; outcome forecast; overexpression; paracrine; personalized approach; pro-apoptotic protein; protein expression; public health relevance; three dimensional cell culture; tumor; tumor progression

DESCRIPTION (provided by applicant): Ductal carcinoma in situ (DCIS) is the most common type of non-invasive breast cancer diagnosed in women and an immediate precursor to invasive ductal carcinoma (IDC). Current cyto- and histopathological approaches do not accurately predict disease progression, resulting in patients being under-treated or over-treated for DCIS. Our long-term goals are to identify key factors that lead to IDC that will enable the development of a molecular based approach to predict the risk of IDC, and a more tailored approach to treat DCIS. CCL2/CCR2 chemokine receptor signaling in DCIS is a potentially important mechanism for DCIS progression to IDC. CCR2 and its binding ligand CCL2, are overexpressed in breast ductal carcinomas, and correlate with tumor grade and poor patient prognosis. CCL2/CCR2 chemokine signaling is best known for regulating macrophage recruitment. However, knockdown of CCL2 significantly inhibits mammary tumor progression without significantly affecting macrophage recruitment. Overexpression of CCR2 in ductal carcinoma cells enhances cell survival and invasiveness, associated with increased expression of pro-survival and pro-invasive proteins (ALDH1A1) and decreased expression of pro-apoptotic proteins (Htra2). These studies indicate an important role for CCR2 signaling in breast ductal carcinoma progression. To clarify the in vivo role of CCL2/CCR2 signaling, we have developed novel xenograft models through mouse Mammary intraductal injection (MIND) of human breast carcinoma cell lines, Sum225 and DCIS.com. The Sum225 MIND model form stable non-invasive lesions, while DCIS lesions progress to IDC in the DCIS.com MIND model. Compared to conventional transplant models, these MIND models more closely mimic the biology of DCIS found in patients. Compared to Sum225 lesions, DCIS.com lesions show higher expression of CCL2, CCR2 and ALDH1A1 proteins and lower expression of Htra2, correlating with progression to invasive carcinoma. Based on preliminary studies, we hypothesize that CCL2/CCR2 signaling in ductal carcinoma cells enhances progression of DCIS to IDC. Aim 1: To characterize the role of autocrine CCL2/CCR2 signaling in ductal carcinoma cells during DCIS by analyzing effects of CCR2 overexpression and knockdown in MIND models, 3D cell culture models and biochemical approaches, using primary and established breast cancer cell lines. Aim 2: To determine the functional contribution of stromal derived CCL2 on CCR2 mediated progression of DCIS, by analyzing the effects of CCL2 derived from fibroblasts on DCIS progression, using subrenal graft transplant models, MIND models, 3D cell culture models and biochemical approaches, using primary and established breast cancer cell lines. Through a multiple PI plan, the objective is to determine how CCL2/CCR2 signaling regulates expression of pro-invasive and pro-survival factors (ALDH1A1) and pro-apoptotic factors (Htra2) to promote ductal carcinoma progression. These studies will reveal mechanistic insight into the role of chemokine receptor signaling in DCIS, with important therapeutic implications. .

描述（由申请人提供）：原位导管癌（DCIS）是在女性中诊断出的最常见的非侵入性乳腺癌类型，也是侵入性导管癌（IDC）的直接前体。当前的细胞病理学方法和组织病理学方法无法准确预测疾病的进展，导致患者在DCIS治疗或治疗过度治疗。我们的长期目标是确定导致IDC的关键因素，从而能够开发基于分子的方法来预测IDC风险，并采用更量身定制的DCIS方法。 DCIS中的CCL2/CCR2趋化因子受体信号传导是DCIS向IDC发展的潜在重要机制。 CCR2及其结合配体CCL2在乳腺导管癌中过表达，并且与肿瘤级和患者预后不良相关。 CCL2/CCR2趋化因子信号传导以调节巨噬细胞募集而闻名。然而，CCL2的敲低显着抑制乳腺肿瘤的进展，而不会显着影响巨噬细胞的募集。 CCR2在导管癌细胞中的过表达增强了细胞的存活和侵袭性，与促寿使和促侵入性蛋白的表达增加有关（ALDH1A1）和促凋亡蛋白的表达降低（HTRA2）。这些研究表明CCR2信号在乳腺导管癌进展中的重要作用。为了阐明CCL2/CCR2信号的体内作用，我们通过人类乳腺癌细胞系，sum225和dcis.com的小鼠乳腺内导管内注射（思维）开发了新型的异种移植模型。 SUM225思维模型形成稳定的非侵入性病变，而DCIS病变在DCIS.com Mind Model中发展为IDC。与常规的移植模型相比，这些思维模型更加亲密地模仿患者中发现的DCI的生物学。与SUM225病变相比，DCIS.com病变显示CCL2，CCR2和ALDH1A1蛋白的表达更高，HTRA2的表达较低，与侵入性癌的发展相关。基于初步研究，我们假设导管癌细胞中的CCL2/CCR2信号传导增强了DCIS向IDC的进展。目标1：通过使用原发性和既定的乳腺癌细胞中CCR2的过表达和敲低的作用来表征DCIS在DCIS期间自分泌CCL2/CCR2信号传导在DCIS期间的导管癌细胞中的作用，并使用原发性和既定的乳腺癌细胞在心理模型，3D细胞培养模型和生化方法中的作用。目的2：通过分析使用下肾移植物模型，心理模型，3D细胞培养模型和生物化学模型和生物化学模型，使用初级和建立的乳腺癌细胞细胞系列，使用肾上腺移植物模型，使用肾上腺培养物模型和生物化学方法来确定源自成纤维细胞对DCIS进展的CCL2的影响，以确定基质衍生的CCL2对CCR2介导的DCI的功能贡献。通过多个PI计划，目的是确定CCL2/CCR2信号传导如何调节促侵入性和促生物学因子（ALDH1A1）和促凋亡因子（HTRA2）的表达以促进导管癌的进展。这些研究将揭示对趋化因子受体信号在DCI中的作用的机理洞察力，具有重要的治疗意义。 。