SFRP2 and NFAT are therapeutic targets in angiosarcoma

SFRP2 和 NFAT 是血管肉瘤的治疗靶点

基本信息

批准号：
8267128
负责人：
Nancy Demore
金额：
$ 29.79万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8267128
关键词：
Angiogenesis Inducing Agents Angiogenesis Inhibitors Angiogenic Factor Antibodies Apoptosis Binding Biological Assay Biology Blood Vessels Blood capillaries Calcineurin Calcineurin inhibitor Calmodulin Cell Cycle Progression Cell Nucleus Cell physiology Cells Clinical Trials Complex Cytoplasm Data Diagnosis Disease Dose Doxorubicin Drug Combinations Drug Kinetics Endothelial Cells Endothelium Equilibrium FDA approved FK506 Future Genes Genetic Transcription Genomics Graft Rejection Growth Hemangiosarcoma Human Hypoxia Immunohistochemistry Immunosuppressive Agents In Vitro Knowledge Laboratories Lead Ligands Lymphocyte Malignant - descriptor Malignant Neoplasms Maximum Tolerated Dose Mediating Molecular Molecular Target Monoclonal Antibodies Mus Nuclear Nuclear Translocation Nude Mice Organ Transplantation Paclitaxel Paraffin Embedding Pathway interactions Patients Pharmaceutical Preparations Phase Phase II Clinical Trials Phosphoric Monoester Hydrolases Play Progression-Free Survivals Protein-Serine-Threonine Kinases Proteins Publishing Regimen Role Signal Pathway Signal Transduction Solid Neoplasm Staining method Stains Tacrolimus Tacrolimus Binding Protein 1A Testing Time Toxic effect Tube Tumor Angiogenesis Tumor Volume Unresectable Vascular Breast Neoplasm Vascular Endothelial Growth Factors Xenograft procedure angiogenesis arm base bevacizumab calcineurin phosphatase capillary cell growth cell motility chemotherapy chorioallantoic membrane design drug efficacy efficacy testing human SFRP4 protein improved in vivo laser capture microdissection matrigel migration mortality new growth novel outcome forecast overexpression polyclonal antibody prevent receptor receptor binding response therapeutic target transcription factor transcription factor NF-AT c3 tumor tumor growth

项目摘要

Angiosarcoma is a biologically aggressive vascular malignancy with a high metastatic potential and subsequent mortality(1). At the time of diagnosis of angiosarcoma, 10¿ 25% of patients already have metastatic disease. The 2- and 5-year overall survival for all patients with angiosarcoma is 50 and 30%, respectively, with a median overall survival of 24 months. For unresectable angiosarcoma, doxorubicin based regimens yield progression-free survival of 5 months, and paclitaxel achieves a progression-free survival of 7 months(2). Therefore there is a desperate need for novel therapies to improve survival in patients with this highly lethal disease. Our laboratory has recently discovered a novel signaling pathway responsible for angiosarcoma growth. While conducting genomic profiling of breast tumor vascular cells compared to normal vessels obtained by laser capture microdissection, we identified secreted frizzled related protein 2 (SFRP2) as a gene with increased expression in tumor endothelium. SFRP2 is a 33kd secreted protein involved in Wnt signaling. Our preliminary data shows that SFRP2 protein is present in 9/9 human angiosarcomas by IHC, and stimulates angiogenesis on the chick chorioallantoic membrane, induces endothelial cell migration and tube formation, and protects against hypoxia induced apoptosis. Silencing of SFRP2 in SVR angiosarcoma cells resulted in inhibition of tube formation, and stimulation of endothelial cells with SFRP2 resulted in an increase in nuclear NFATc3. NFAT is a transcription factor that plays a critical role in mediating angiogenic responses(3;4). NFAT nuclear localization is dependent on a dynamic import-export balance between the activity of the Ca2+/calmodulin-dependent phosphatase, calcineurin, and the activity of serine/threonine kinases(20). NFAT cannot normally enter the nucleus until it is dephosphorylated, but can be activated by calcineurin. Activated calcineurin dephosphorylates NFAT, which then translocates from the cytoplasm to the nucleus and results in transcription of genes involved in cell growth, differentiation, and cell cycle progression. Tacrolimus (FK506) is an immunosuppressive drug that binds to the immunophlin FKBP12 in lymphocytes. The FK506-FKBP12 complex associates with calcineurin and inhibits its phosphatase activity, which inhibits nuclear translocation of NFAT(5). Our preliminary data shows that tacrolimus inhibits angiosarcoma tube formation in vitro, and systemic administration inhibits the growth of the SVR angiosarcoma xenograft in nude mice by 46% at 20 days. Taken together, our data leads us to propose the following hypothesis: Blockade of SFRP2 will decrease angiogenesis and angiosarcoma growth and will synergistically improve tumor regression in combination with both chemotherapy and antiangiogenic therapy. The objective of this study is to show the efficacy of these drugs in angiosarcoma alone, and in combination with chemotherapy and antiangiogenic therapy, and to elucidate their molecular mechanism in inhibiting tumor growth. Successful completion of the project will lead to clinical trials of these agents in patients with angiosarcoma in future studies.

血管肉瘤是一种生物学上攻击性的血管恶性肿瘤，具有高转移潜力和随后的死亡率（1）。在诊断为血管肉瘤时，有102％的患者已经患有转移性疾病。所有血管肉瘤患者的2年和5年总生存期为50％和30％，中位总生存期分别为24个月。对于不可切除的血管肉瘤，基于阿霉素方案的无进展生存率为5个月，紫杉醇可实现无进展的生存率为7 月（2）。因此，迫切需要新的疗法来改善此患者的生存率高致命的疾病。我们的实验室最近发现了一个新的信号途径血管肉瘤的生长。在进行乳腺肿瘤血管细胞基因组分析的同时通过激光捕获微分解获得的血管，我们确定了分泌的毛躁相关蛋白2（SFRP2）为在肿瘤内皮中表达增加的基因。 SFRP2是参与Wnt的33KD分泌蛋白信号。我们的初步数据表明，SFRP2蛋白是由IHC在9/9的人类肉瘤中呈现的，而SFRP2蛋白则显示刺激鸡绒毛膜膜上的血管生成，诱导内皮细胞迁移和管形成，预防缺氧诱导的凋亡。 SVR血管肉瘤细胞中SFRP2的沉默导致抑制管形成，并用SFRP2刺激内皮细胞导致增加在核NFATC3中。 NFAT是转录因子，在介导血管生成中起着至关重要的作用响应（3; 4）。 NFAT核定位取决于动态进出口的平衡 Ca2+/钙调蛋白依赖性磷酸酶，钙调蛋白和丝氨酸/苏氨酸的活性激酶（20）。 NFAT直到被脱磷酸化之前通常无法进入核，但可以通过钙调神经素。激活的钙调神经蛋白去磷酸化NFAT，然后从细胞质转移到细胞核和导致涉及细胞生长，分化和细胞周期进程的基因转录。他克莫司（FK506）是一种免疫抑制性药物，与淋巴细胞中的免疫链接FKBP12结合。 FK506-FKBP12复合物与钙调神经酶相关并抑制其磷酸酶活性，从而抑制 NFAT的核易位（5）。我们的初步数据表明他克莫司抑制血管肉瘤管体外形成，全身给药抑制了裸体中SVR血管肉瘤异种移植的生长小鼠在20天时的小鼠46％。综上所述，我们的数据导致我们提出以下假设：封锁 SFRP2将减少血管生成和血管肉瘤的生长，并协同改善肿瘤回归结合化学疗法和抗血管生成疗法。这项研究的目的是表明这些药物仅在血管肉瘤中的功效，并结合化学疗法和抗血管生成治疗，并在抑制肿瘤生长中阐明其分子机制。成功完成项目将导致这些药物在未来研究中的血管肉瘤患者中进行这些药物的临床试验。