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: 9134843
• 负责人: Minae Mure
• 金额: $ 28.7万
• 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/9134843
• 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; LOXL2 gene; 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; 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; LOXL2 gene; 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.

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属于赖氨酸氧化酶（LOX）家族，该家族由赖氨酸酪氨酸（LTQ）依赖性铜胺氧化酶组成。通常认为LOXL2通过其胺氧化酶活性催化ECM僵硬。但是，最近还提出了LOXL2的细胞内功能。迄今为止，尚无LOXL2或LOX家族成员的任何结构，并且已经进行了非常有限的LOXL2生化和生物学研究。我们已经发现，核（未糖基化的）LOXL2会诱导乳腺癌细胞的上皮to-中质质转变（EMT，转移的第一步，并促进细胞的增殖和浸润比分泌的（N-糖基化）LOXL2在体外更有效。我们还发现，分泌的LOXL2经历核转移以诱导EMT。我们的中心假设是，未糖基化的LOXL2定位于细胞核，并且通过以胺氧化酶活性依赖性方式稳定Snail1转录因子，可以通过稳定Snail1转录因子诱导EMT和侵袭。因此，我们希望制定抑制LOXL2的生产，核积累和/或活性的策略，这些策略有可能发展为表达核LOXL2的癌症的靶向疗法。在此提案中，我们将定义LOXL2的翻译后修饰（PTM），并在指导LOXL2在乳腺癌转移/侵袭中指导LOXL2的不同分子功能中的作用。成功完成拟议的研究将为LOXL2的结构函数相关性提供首次实质性见解。最终，这项研究的成功将为靶向疗法的设计提供了基础样乳腺癌的靶向疗法的设计，表达核LOXL2水平升高。