Extracellular Matrix Remodeling and the Control of Invasive Lobular Carcinoma Progression

细胞外基质重塑和侵袭性小叶癌进展的控制

• 批准号: 10677993
• 负责人: Adam Wallace Olson
• 金额: $ 4.04万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677993
• 关键词: Accounting; Address; Affect; Alleles; Automobile Driving; Behavior; Binding; Biological; Carcinoma; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen; Collagen Type I; Complex; Cre driver; Data; Deposition; Diffuse; E-Cadherin; Early Diagnosis; Engraftment; Extracellular Matrix; Fibroblasts; Gene Expression Profile; Genetic Transcription; Growth; Housing; Human; Individual; Invaded; Invasive Lesion; Laboratories; Left; Lobular Carcinoma; LoxP-flanked allele; MMP14 gene; Malignant Epithelial Cell; Malignant Neoplasms; Matrix Metalloproteinases; Mechanics; Mediating; Membrane; Mesenchymal; Modeling; Molecular; Morbidity - disease rate; Mus; Mutation; Nature; Neoplasm Metastasis; Neoplasms; Normal Cell; PIK3CG gene; Patients; Pattern; Peptide Hydrolases; Play; Process; Radiation therapy; Research; Resistance; Role; Signal Transduction; Stromal Cells; System; Tissues; Xenograft Model; arm; cancer cell; cell type; chemotherapy; density; ductal breast carcinoma; experimental study; improved; in vivo; individualized medicine; inducible Cre; insight; malignant breast neoplasm; mammary epithelium; mechanotransduction; migration; mortality; mouse model; neoplastic cell; new therapeutic target; novel; programs; recruit; single-cell RNA sequencing; therapeutic target; transcriptomics; tumor; tumor progression; tumorigenic

Proposal Summary Invasive lobular carcinoma (ILC) is the second most common subtype of breast cancer, and accounts for ~10% of all breast cancers. ILCs, highlighted by lost E-cadherin expression and diffuse, non-cohesive growth, are highly invasive, traversing the surrounding extracellular matrix (ECM) in a streaking pattern of single cells. While ILCs are relatively slow growing, preclusion of early detection, resistance to common chemo- and radiotherapies, and their highly invasive nature make them exceptionally difficult to eradicate, leading to significant patient morbidity and mortality. For ILCs to invade and metastasize, they must navigate the type I collagen-rich ECM surrounding the mammary epithelia, a process recently suggested to require proteolytic degradation, despite prior notions of proteolytic-independent migration. A specific membrane-bound matrix metalloproteinase capable of degrading type I collagen, MT1-MMP, has further been demonstrated to play a particularly crucial role in carcinoma invasion. To this end, MT1-MMP has been shown to be expressed in both ILC cells and surrounding cancer-associated fibroblasts (CAFs). Inversely, ILCs display an extensive accumulation of type I collagen deposition directly surrounding invasive lesions. However, it remains the subject of debate whether this increase in local ECM density confers pro- or anti-tumorigenic effects on ILCs and whether ECM deposition is primarily ILC cell- or CAF-derived. Further complicating the effects of ILC cell-ECM interactions in vivo, changes in MT1- MMP expression and ECM composition, in other systems, have been demonstrated to induce changes in transcription profiles. Therefore, this proposal aims to individually address each arm of the MT1-MMP/collagen axis characterizing its effects on ILC growth, invasion, and metastasis in vivo, in a manner specific to ILC cells and their surrounding CAFs. We hypothesize that both ILC cell- and CAF-derived MT1-MMP activity and ECM deposition influence ILC progression through physical remodeling of the ECM as well as via tumor-ECM induced transcriptomic changes. Experiments in Aim 1 will assess the role of ILC cell- and CAF-derived MT1-MMP during ILC progression using a combination of spontaneous and engrafted ILC models housing an Mt1-mmp floxed allele in mice and CRISPR-mediated MT1-MMP deletion in human ILC cells. Meanwhile, Aim 2 will separately assess the effects of altered ECM deposition in a cell-type specific manner, similarly, using spontaneous and engrafted ILC mouse models where a floxed Col1a1 allele has been inserted to disrupt collagen deposition by either carcinoma cells or the surrounding stroma. The experiments in Aims 1 & 2 will be further interrogated using advanced transcriptomic approaches including bulk and single-cell RNA sequencing to unbiasedly investigate effects on both targeted and surrounding cells. Collectively, this proposal aims to characterize the influences of the MT1-MMP/collagen axis on ILC progression and its associated transcriptional programs. Such insights will improve our understanding of the mechanisms underlying ILC growth, invasion, and metastasis, while simultaneously identifying potential targets for the tailored treatment of ILC in affected patients.

提案摘要 侵入性小叶癌（ILC）是乳腺癌的第二常见亚型，占约10％ 在所有乳腺癌中。 ILC是由丧失的电子钙粘蛋白表达和弥漫性，非粘性生长突出的，是 高度侵入性，以单个细胞的条纹模式横穿周围的细胞外基质（ECM）。尽管 ILC的生长相对较慢，对早期检测的排除，对常见化学疗法和放射性疗法的抗性， 它们的高度侵入性使他们难以消除，导致了重要的患者 发病率和死亡率。为了使ILC入侵和转移，他们必须导航I型胶原蛋白丰富的ECM 围绕乳腺上皮，尽管 蛋白水解独立迁移的先前概念。特定膜结合的基质金属蛋白酶 MT1-MMP的I型I型胶原蛋白的降解，进一步被证明在 癌入侵。为此，已显示MT1-MMP在ILC细胞和周围都表达 癌症相关的成纤维细胞（CAF）。成反比，ILC显示了I型胶原蛋白的广泛积累 直接围绕侵入性病变的沉积。但是，这仍然是辩论的主题 在局部ECM密度中，对ILC的促或抗氧化作用以及ECM沉积是否主要是 ILC细胞或CAF衍生。进一步使ILC细胞ECM相互作用在体内的影响，MT1-的变化变化 在其他系统中，MMP表达和ECM组成已被证明可以引起变化 转录曲线。因此，该建议旨在单独解决MT1-MMP/胶原蛋白的每个臂 轴心表征其对体内ILC生长，侵袭和转移的影响，以ILC细胞为特定的方式 和他们周围的咖啡馆。我们假设ILC细胞和CAF衍生的MT1-MMP活性和ECM都 沉积通过ECM的物理重塑以及通过肿瘤诱导的物理重塑影响ILC的进展 转录组变化。 AIM 1中的实验将评估ILC细胞和CAF衍生的MT1-MMP的作用 使用自发和植入的ILC模型组合的ILC进展 小鼠和CRISPR介导的MT1-MMP缺失的等位基因在人ILC细胞中的缺失。同时，AIM 2将分别 同样，使用自发和 植入了ILC小鼠模型，其中已插入了Floxed Col1a1等位基因以破坏胶原蛋白沉积 癌细胞或周围基质。目标1和2中的实验将进一步审问 使用先进的转录组方法，包括散装和单细胞RNA测序 研究对靶向细胞和周围细胞的影响。总的来说，该建议旨在表征 MT1-MMP/胶原蛋白轴对ILC进展及其相关转录程序的影响。这样的 见解将提高我们对ILC增长，入侵和转移基础机制的理解， 同时确定了受影响患者的ILC量身定制治疗的潜在靶标。