Understanding the roles of PTM's in modulating molecular functions of lysyl oxidase-like 2 in breast cancer cells

了解 PTM 在调节乳腺癌细胞赖氨酰氧化酶样 2 分子功能中的作用

• 批准号: 8931006
• 负责人: Minae Mure
• 金额: $ 28.72万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-22 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8931006
• 关键词: Animal Model; Antibodies; Biochemical; Biological; Biological Markers; Breast Cancer Cell; Breast Cancer Patient; Breast cancer metastasis; Cell Nucleus; Cell Proliferation; Cells; Cellular biology; Collagen; Distant; Extracellular Matrix; Family; Female; Fibroblasts; Glycobiology; Goals; In Vitro; Kansas; Kinetics; Knowledge; Lysine; Mass Spectrum Analysis; Metastatic breast cancer; Methodology; Methods; Modification; Molecular; N-terminal; Neoplasm Metastasis; Nuclear; Nuclear Translocation; Patients; Pharmaceutical Preparations; Polysaccharides; Post-Translational Protein Processing; Procedures; Process; Production; Property; Protein Chemistry; Protein Family; Protein Isoforms; Protein-Lysine 6-Oxidase; Proteins; Research; Role; Scavenger Receptor Cysteine-Rich Domain; Series; Side; Staging; Structure; Substrate Specificity; Survival Rate; Testing; Tissues; Universities; aggressive therapy; amine oxidase; base; cancer cell; crosslink; demethylation; design; diamino oxhydrase; epithelial to mesenchymal transition; insight; malignant breast neoplasm; member; new therapeutic target; public health relevance; success; targeted cancer therapy; targeted treatment; therapeutic target; tissue culture; transcription factor; tumor progression; uptake

DESCRIPTION (provided by applicant): Lysyl oxidase-like 2 (LOXL2) is anticipated to be a promising novel therapeutic target in basal- like breast cancers, because it is highly upregulated in these cancer cells and tissues, and because inhibition of the production of LOXL2 by shRNAs or treatment with its specific antibody have been shown to retard tumor progression and metastasis/invasion in tissue culture and in animal models. LOXL2 belongs to the lysyl oxidase (LOX) family, which consists of lysine tyrosylquinone (LTQ)-dependent copper amine oxidases. LOXL2 is generally considered to catalyze ECM stiffening by its amine oxidase activity. However, intracellular functions of LOXL2 have also recently been proposed. To date, there are no structures for LOXL2 or any member of the LOX family, and very limited fundamental biochemical and biological study of LOXL2 has been conducted. We have discovered that nuclear (unglycosylated) LOXL2 induces epithelial- to-mesenchymal transition (EMT, the first step of metastasis) of breast cancer cells and promotes cell proliferation and invasion much more effectively than secreted (N-glycosylated) LOXL2 does in vitro. We also found that secreted LOXL2 undergoes nuclear-translocation to induce EMT. Our central hypothesis is that unglycosylated LOXL2 localizes to the nucleus, and there induces EMT and invasion by stabilizing Snail1 transcription factor in an amine oxidase activity-dependent fashion. Therefore, we wish to develop strategies to inhibit the production, nuclear accumulation, and/or activity of LOXL2, which could potentially be developed into a targeted therapy for cancers expressing nuclear LOXL2. In this proposal we will define the post-translational modifications (PTMs) of LOXL2 and decipher their roles in directing distinct molecular functions of LOXL2 in breast cancer metastasis/invasion. Successful completion of the proposed studies will provide the first substantial insight into the structure- function correlation of LOXL2. Ultimately, the success of this study will inform the design of targeted therapies for a subtype of basal-like breast cancers expressing elevated levels of nuclear LOXL2.

描述（由申请人提供）：赖氨酰氧化酶样 2 (LOXL2) 预计将成为基底样乳腺癌的一个有前途的新型治疗靶点，因为它在这些癌细胞和组织中高度上调，并且因为通过 shRNA 抑制 LOXL2 的产生或用其特异性抗体治疗已被证明可以在组织培养和动物模型中延缓肿瘤进展和转移/侵袭。 LOXL2 属于赖氨酰氧化酶 (LOX) 家族，该家族由依赖赖氨酸酪氨酰醌 (LTQ) 的铜胺氧化酶组成。 LOXL2 通常被认为通过其胺氧化酶活性来催化 ECM 硬化。然而，LOXL2 的细胞内功能最近也被提出。迄今为止，还没有 LOXL2 或 LOX 家族任何成员的结构，并且对 LOXL2 进行的基础生化和生物学研究非常有限。我们发现核（未糖基化）LOXL2 在体外比分泌型（N-糖基化）LOXL2 更有效地诱导乳腺癌细胞上皮间质转化（EMT，转移的第一步）并促进细胞增殖和侵袭。我们还发现分泌的 LOXL2 经历核转位以诱导 EMT。我们的中心假设是，未糖基化的 LOXL2 定位于细胞核，并通过以胺氧化酶活性依赖的方式稳定 Snail1 转录因子来诱导 EMT 和侵袭。因此，我们希望开发抑制LOXL2的产生、核积累和/或活性的策略，这有可能开发成表达核LOXL2的癌症的靶向治疗。在本提案中，我们将定义 LOXL2 的翻译后修饰 (PTM)，并破译它们在指导 LOXL2 在乳腺癌转移/侵袭中的独特分子功能中的作用。成功完成拟议的研究将为 LOXL2 的结构-功能相关性提供第一个实质性的见解。最终，这项研究的成功将为针对核 LOXL2 表达水平升高的基底样乳腺癌亚型的靶向治疗设计提供信息。