Mechanism of SPARC peptide FSEC inhibition of angiogenesis in neuroblastoma

SPARC肽FSEC抑制神经母细胞瘤血管生成的机制

基本信息

批准号：
8512433
负责人：
SUSAN L. COHN
金额：
$ 23.7万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8512433
关键词：
Affinity Chromatography Amino Acids Angiogenesis Inhibition Binding Biological Blood Vessels C-terminal Cells Child Clinical Cysteine Development Diazomethane Drug Targeting EGF gene Endothelial Cells Engineering Extracellular Matrix Extracellular Matrix Proteins Follistatin Goals Grant Growth Human Incubated Lead Length Malignant - descriptor Malignant Childhood Neoplasm Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Maps Mass Spectrum Analysis Migration Assay Modality Neuroblastoma Outcome Pancreas Patients Peptide Synthesis Peptides Phase Plant Resins Pre-Clinical Model Property Proteins Proteomics Recurrent disease Research Risk Schwann Cells Solid Streptavidin Structure Technology Testing Therapeutic Tumor Suppressor Proteins Tumor Tissue Xenograft procedure analog angiogenesis base cancer cell cancer therapy crosslink design effective therapy high risk improved insight malignant breast neoplasm mortality neoplastic neoplastic cell neuroblast new technology new therapeutic target novel novel therapeutics pre-clinical preclinical study public health relevance tumor tumor growth tumor microenvironment tumor xenograft ultraviolet irradiation

Affinity Chromatography Amino Acids Angiogenesis Inhibition Binding Biological Blood Vessels C-terminal Cells Child Clinical Cysteine Development Diazomethane Drug Targeting EGF gene Endothelial Cells Engineering Extracellular Matrix Extracellular Matrix Proteins Follistatin Goals Grant Growth Human Incubated Lead Length Malignant - descriptor Malignant Childhood Neoplasm Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Maps Mass Spectrum Analysis Migration Assay Modality Neuroblastoma Outcome Pancreas Patients Peptide Synthesis Peptides Phase Plant Resins Pre-Clinical Model Property Proteins Proteomics Recurrent disease Research Risk Schwann Cells Solid Streptavidin Structure Technology Testing Therapeutic Tumor Suppressor Proteins Tumor Tissue Xenograft procedure analog angiogenesis base cancer cell cancer therapy crosslink design effective therapy high risk improved insight malignant breast neoplasm mortality neoplastic neoplastic cell neuroblast new technology new therapeutic target novel novel therapeutics pre-clinical preclinical study public health relevance tumor tumor growth tumor microenvironment tumor xenograft ultraviolet irradiation

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项目摘要

DESCRIPTION (provided by applicant): Despite great progress in the treatment of pediatric cancers, mortality among children with high-risk neuroblastoma remains high, with nearly 60% of the patients developing recurrent disease that is incurable with existing therapies. Traditional cancer treatment use drugs that target the neoplastic cells. However, it is now established that the non-neoplastic components of the tumor (i.e., the tumor stroma/microenvironment) can either promote or inhibit tumor growth depending upon its cellular composition and the organization of the extracellular matrix. Matricellular proteins are natural regulators of the interactions between cancer cells and their microenvironment. We and others have shown that the matricellular protein Secreted Protein Acidic and Rich in Cysteine (SPARC) regulates the assembly and composition of the extracellular matrix, blocks angiogenesis, and inhibits the growth of neuroblastoma and other neoplastic tumors. We have conducted structure-functional studies by testing the anti-angiogenic activity of SPARC constructs with deletions in the 3 major domains, and a number of SPARC peptides. We have further demonstrated that peptide FS-E (22 amino acids), which corresponds to an EGF module of a follistatin domain, has anti-tumor activity. Most recently, we have designed a smaller SPARC peptide (11 amino acids), FSEC, corresponding to the C-terminal part of FS- E, and have demonstrated that it also has potent anti-angiogenic activity and inhibits the growth of neuroblastoma in preclinical models. Further, our preliminary studies show that FSEC inhibits the growth of pancreatic, lung, and breast cancer xenografts, indicating that FSEC may have therapeutic activity in a broad range of tumor types. The specific aim of this study is to elucidate the mechanisms by which peptide FSEC inhibits angiogenesis and neuroblastoma tumor growth by identifying FSEC-binding partners through: 1) cross-linking FSEC engineered with a diazirine motif with human neuroblastoma tumor tissue proteins; 2) purifying binding partners by affinity chromatography, and 3) identifying protein partners with mass-spectrometry. We will use a systemic proteomic approach to isolate the proteins that interact with FSEC. To engineer FSEC probes, we will use new technology that incorporates a highly reactive diazirine group into synthesized peptide probes. These probes will be incubated with human neuroblastoma tumor tissue, and cross-linking will be performed under UV-irradiation. Binding partners will be purified by affinity chromatography and identified by mass-spectrometry. Identification of the proteins that interact with FSEC will lead to a better understanding of how FSEC inhibits angiogenesis and tumor growth, and will provide insight for the development of novel therapeutics that target non-neoplastic cells in the tumor by creating a non- permissive microenvironment. The long-term goal of our research is to improve the outcome of children with high-risk neuroblastoma by developing effective therapeutic strategies that target the microenvironment.

描述（由申请人提供）：尽管在儿科癌症治疗方面取得了长足的进展，但具有高危神经母细胞瘤的儿童的死亡率仍然很高，其中近60％的患者患有复发性疾病，而现有疗法则无法治愈。传统的癌症治疗使用针对肿瘤细胞的药物。但是，现在已经确定肿瘤的非肿瘤成分（即肿瘤基质/微环境）可以根据其细胞组成和细胞外基质的组织来促进或抑制肿瘤的生长。母细胞蛋白是癌细胞及其微环境之间相互作用的自然调节剂。我们和其他人表明，女生分泌的蛋白质酸性并富含半胱氨酸（SPARC）调节细胞外基质的组成和组成，阻断血管生成，并抑制神经母细胞瘤和其他肿瘤肿瘤的生长。我们通过测试3个主要结构域中缺失的SPARC构建体的抗血管生成活性和许多SPARC肽进行了结构功能研究。我们进一步证明，对应于Follistatin结构域的EGF模块的肽FS-E（22个氨基酸）具有抗肿瘤活性。最近，我们设计了与FS-E的C末端部分相对应的较小的SPARC肽（11个氨基酸），FSEC，并证明它也具有有效的抗血管生成活性并抑制史努比模型中神经母细胞瘤的生长。此外，我们的初步研究表明，FSEC抑制胰腺癌，肺和乳腺癌异种移植物的生长，表明FSEC可能在广泛的肿瘤类型中具有治疗活性。这项研究的具体目的是阐明肽FSEC抑制血管生成和神经母细胞瘤肿瘤生长的机制，通过以下方式鉴定FSEC结合伴侣：1）通过以下：1）用人神经母细胞瘤肿瘤组织蛋白进行十字齐瑞碱基序的交联FSEC; 2）通过亲和力色谱法净化结合伙伴，3）识别具有质谱法的蛋白质合作伙伴。我们将使用一种系统性的蛋白质组学方法来隔离与FSEC相互作用的蛋白质。对于工程FSEC探针，我们将使用将高度反应性重氮嗪组纳入合成肽探针中的新技术。这些探针将与人类神经母细胞瘤肿瘤组织一起孵育，并将在紫外线辐射下进行交联。结合伴侣将通过亲和力色谱法纯化，并通过质谱法鉴定。鉴定与FSEC相互作用的蛋白质将更好地理解FSEC如何抑制血管生成和肿瘤生长，并将为新的疗法的发展提供洞察力，这些新疗法的发展，这些疗法靶向肿瘤中的非肿瘤细胞，通过创建非渗透性微环境。我们研究的长期目标是通过制定针对微环境的有效治疗策略来改善具有高危神经母细胞瘤的儿童的结果。