Cellular or Extracellular Targeting of Lysyl Oxidase Propeptide for Oral Cancer

赖氨酰氧化酶前肽的细胞或细胞外靶向治疗口腔癌

• 批准号: 8768580
• 负责人: PHILIP C TRACKMAN
• 金额: $ 24.56万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8768580
• 关键词: Anabolism; Apoptosis; Attention; Binding; Cancer Cell Growth; Cancer Model; Cancer cell line; Cell Proliferation; Cell surface; Cells; Cessation of life; Chimeric Proteins; Collagen; Data; Diagnosis; Drug Formulations; Ectopic Expression; Effectiveness; Elastin; Engineering; Enzyme Precursors; Enzymes; Extracellular Matrix; Extracellular Matrix Proteins; Extracellular Protein; FGFR1 gene; Fc domain; Fibroblast Growth Factor 2; Fibroblast Growth Factor Receptors; Fostering; Genes; Growth; Heat-Shock Proteins 70; Human; IgG4; Implant; In Vitro; Incidence; Knowledge; Laboratories; Location; Malignant Neoplasms; Mediating; Modeling; Modification; Mouth Neoplasms; Mus; Oral; Palmitic Acids; Pathway interactions; Patients; Phenotype; Process; Property; Protein-Lysine 6-Oxidase; Proteins; Proteolytic Processing; Publishing; RAS inhibition; Recombinants; Relative (related person); Research; Serum; Signal Transduction; Site; Testing; Therapeutic; Tongue; Tongue Neoplasms; Toxic effect; Tubulin; Tumor Markers; Tumor Suppressor Proteins; Work; anti-cancer therapeutic; base; cancer cell; cell growth; cell transformation; chemotherapeutic agent; crosslink; design; extracellular; improved; in vivo; insight; malignant mouth neoplasm; mouth squamous cell carcinoma; neoplastic cell; oral tissue; outcome forecast; procollagen C-endopeptidase; protein aminoacid sequence; public health relevance; receptor binding; treatment strategy; tumor; tumor growth; tumorigenesis; uptake

DESCRIPTION (provided by applicant): Oral cancer incidence is rising and is the now the 8th most common form of cancer. Prognosis is poor; at least 50% of patients die within five years of diagnosis. Lysyl oxidase (LOX) is a critically important extracellular matrix protein and enzyme. LOX is synthesized as a 50 kDa pre-proenzyme (Pro-LOX) that is secreted, and then processed extracellularly by procollagen C-proteinases to form active 30 kDa lysyl oxidase enzyme, and the 18 kDa lysyl oxidase propeptide (LOX-PP). The tumor suppressor activity originally attributed to LOX enzyme depends instead on LOX-PP. LOX-PP is a naturally occurring extracellular matrix protein, and likely has limited toxicity while having important anti-cancer activities. Different regions of LOX-PP bind to different targets in its inhibition of RAS-dependen cancer pathways. Some targets are intracellular (c-RAF and tubulin for example), while others are extracellular (FGFR1, for example). It is currently unknown which location and targets of LOX-PP are most important for its tumor suppressor activity. Knowledge regarding the functional location (intracellular or extracellular) is important for formulation of therapeutics based on rLOX-PP. The proposed research will establish which location of rLOX-PP is functional in its ability to inhibit oral cancer phenotype in vitro (Aim 1), and orthotopic oral tuor growth in mice in vivo (Aim 2). Particular attention will be paid to the HSP70 cancer cell marker which is a rLOX-PP binding partner and occurs both extracellularly and intracellularly and participates in RAS-dependent signal transduction and cell transformation. Fusion protein derivatives which are designed to either remain extracellular by fusion with the Fc-domain of IgG4, or are designed for increased cell uptake by fusion with cell penetrating peptide sequences, or palmitic acid, will be created and tested for anti-cancer activities and extracellular/intracellular location in vitro. These rLOX-PP derivatives are expected to have increased in vivo stability. The most active derivative will be systemically administered to mice i which tongue orthotopic tumors have been implanted using UMSCC2 (aggressive) and CAL 27 (less aggressive) oral cancer cell lines and tumor growth and expression of tumor markers and active RAS activity effectors will be determined. It is expected that at least one rLOX-PP derivative with both increased stability and anti-oral tumor growth effectiveness will be identifie. The importance of extracellular compared to intracellular interactions of the HSP70-binding domain of rLOX-PP in particular will be evaluated. Important information will be gained to further refine the design of anti-tumor molecules.

描述（申请人提供）：口腔癌的发病率正在上升，目前是第8大最常见的癌症。预后差；至少50%的患者在确诊后五年内死亡。赖氨酸氧化酶（LOX）是一种重要的细胞外基质蛋白和酶。LOX以50 kDa的前酶（Pro-LOX）的形式分泌合成，然后由前胶原c蛋白酶在细胞外加工形成活性的30 kDa赖氨酸氧化酶和18 kDa赖氨酸氧化酶前肽（LOX- pp）。最初归因于LOX酶的肿瘤抑制活性取决于LOX- pp。LOX-PP是一种天然存在的细胞外基质蛋白，可能具有有限的毒性，但具有重要的抗癌活性。LOX-PP的不同区域在抑制ras依赖的癌症途径中结合不同的靶点。一些靶点是细胞内的（例如c-RAF和微管蛋白），而其他靶点是细胞外的（例如FGFR1）。目前尚不清楚LOX-PP的哪个位置和靶点对其肿瘤抑制活性最重要。关于功能位置（细胞内或细胞外）的知识对于制定基于rLOX-PP的治疗方法非常重要。拟议的研究将确定rLOX-PP的哪个位置在体外抑制口腔癌表型（Aim 1）和小鼠体内原位口腔肿瘤生长（Aim 2）方面发挥作用。我们将特别关注HSP70癌细胞标记物，它是rLOX-PP的结合伙伴，在细胞外和细胞内都存在，参与ras依赖的信号转导和细胞转化。融合蛋白衍生物通过与IgG4的fc结构域融合而保持在细胞外，或者通过与细胞穿透肽序列或棕榈酸融合而增加细胞摄取，将被创建并在体外测试抗癌活性和细胞外/细胞内定位。这些rLOX-PP衍生物有望提高体内稳定性。使用UMSCC2（侵袭性）和CAL 27（侵袭性较低）口腔癌细胞系植入舌原位肿瘤的小鼠，将系统给予最有效的衍生物，并测定肿瘤生长和肿瘤标志物和活性RAS活性效应器的表达。预计将发现至少一种具有更高稳定性和抗口腔肿瘤生长有效性的rLOX-PP衍生物。将特别评估rLOX-PP的hsp70结合域的细胞外相互作用与细胞内相互作用的重要性。将获得进一步完善抗肿瘤分子设计的重要信息。