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 浸润性小叶癌 (ILC) 是乳腺癌第二常见的亚型，约占 10% 所有乳腺癌。 ILC 的特点是 E-钙粘蛋白表达缺失和弥漫性、非粘性生长， 具有高度侵入性，以单细胞的条纹模式穿过周围的细胞外基质（ECM）。尽管 ILC 生长相对缓慢，无法早期检测，对常见的化疗和放疗有抵抗力， 它们的高度侵入性使它们极难根除，导致大量患者死亡 发病率和死亡率。对于 ILC 的侵袭和转移，它们必须导航富含 I 型胶原蛋白的 ECM 围绕乳腺上皮细胞，最近提出一个过程需要蛋白水解降解，尽管 不依赖于蛋白水解的迁移的先前概念。一种特定的膜结合基质金属蛋白酶 降解 I 型胶原蛋白 MT1-MMP 已被进一步证明在 癌侵袭。为此，MT1-MMP 已被证明在 ILC 细胞和周围细胞中表达。 癌症相关成纤维细胞（CAF）。相反，ILC 显示 I 型胶原蛋白的广泛积累 直接沉积在侵袭性病变周围。然而，这种增加是否会增加仍然是一个争论的话题。 局部 ECM 密度对 ILC 具有促肿瘤作用还是抗肿瘤作用，以及 ECM 沉积是否主要是 ILC 细胞或 CAF 衍生。 MT1-的变化使ILC细胞-ECM体内相互作用的影响进一步复杂化 在其他系统中，MMP 表达和 ECM 成分已被证明可引起 转录概况。因此，该提案旨在单独解决 MT1-MMP/胶原蛋白的每个臂 轴以 ILC 细胞特有的方式表征其对体内 ILC 生长、侵袭和转移的影响 及其周边的 CAF。我们假设 ILC 细胞和 CAF 衍生的 MT1-MMP 活性和 ECM 沉积通过 ECM 的物理重塑以及肿瘤 ECM 诱导影响 ILC 进展 转录组变化。目标 1 中的实验将评估 ILC 细胞和 CAF 衍生的 MT1-MMP 在 使用含有 Mt1-mmp floxed 的自发和移植 ILC 模型组合进行 ILC 进展 小鼠中的等位基因和人类 ILC 细胞中 CRISPR 介导的 MT1-MMP 缺失。同时，目标2将分别 类似地，使用自发和 移植的 ILC 小鼠模型，其中插入了 floxed Col1a1 等位基因以破坏胶原蛋白沉积 癌细胞或周围的间质。目标 1 和 2 中的实验将受到进一步质疑 使用先进的转录组学方法，包括批量和单细胞 RNA 测序，以公正地 研究对目标细胞和周围细胞的影响。总的来说，该提案旨在描述 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 生长、侵袭和转移机制的理解， 同时确定针对受影响患者进行 ILC 定制治疗的潜在目标。