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：
7984479
负责人：
Nancy Demore
金额：
$ 30.71万
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/7984479
关键词：
Angiogenesis Inducing Agents Angiogenesis Inhibitors Angiogenic Factor Antibodies Apoptosis Binding Biological Assay Biology Blood Vessels 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 public health relevance receptor receptor binding response therapeutic target transcription factor transcription factor NF-AT c3 tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Angiogenesis is the growth of new capillary blood vessels, and is a critical component of solid tumor growth. We have discovered a novel protein, secreted-frizzled related protein 2, that is overexpressed in human angiosarcoma that is a potent stimulator of angiogenesis. The objective of this proposal is to characterize the molecular and cellular mechanism of this protein on angiosarcoma biology, and evaluate the efficacy of blockade of this signaling pathway on angiosarcoma xenograft growth.

描述（由适用提供）：血管肉瘤是一种具有较高转移潜力和随后死亡率的生物侵袭性血管恶性肿瘤（1）。在诊断为血管肉瘤时，有10 25％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年总生存期分别为50％和30％，中位总生存期为24个月。对于不可切除的血管肉瘤，基于阿霉素的治疗方案的无进展生存期为5个月，紫杉醇可实现7个月的无进展生存期（2）。因此，迫切需要新的疗法来改善这种致命疾病的患者的生存。我们的实验室最近发现了一种负责血管肉瘤生长的新型信号通路。与通过激光捕获显微解剖获得的正常血管相比，对乳腺肿瘤血管细胞进行基因组分析时，我们确定了分泌的毛躁相关蛋白2（SFRP2）为一种基因，在肿瘤森植物中表达增加。 SFRP2是参与WNT信号传导的33KD分泌蛋白。我们的初步数据表明，SFRP2蛋白通过IHC在9/9的人体肉瘤中呈现，并刺激鸡绒毛绒毛膜膜上的血管生成，可诱导内皮细胞迁移和试管形成，并预防低氧诱导的凋亡。 SFRP2在SVR血管肉瘤细胞中的沉默导致导管形成抑制，并用SFRP2刺激内皮细胞导致核NFATC3的增加。 NFAT是转录因子，在介导血管生成反应中起关键作用（3; 4）。 NFAT核定位取决于Ca2+/钙调蛋白依赖性磷酸酶，钙调蛋白的活性与丝氨酸/苏氨酸激酶的活性之间的动态进出口平衡（20）。 NFAT在核脱磷酸化之前通常无法进入核，但可以被钙调蛋白激活。活化的钙调蛋白去磷酸化NFAT，然后从细胞质转移到核，并导致与细胞生长，分化和细胞周期进程有关的基因转录。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，他克莫司在体外抑制了血管肉瘤的形成，全身给药抑制了20天在裸鼠中SVR Angiosarcoma eenographogrogroticon的生长，在20天时就会抑制46％。综上所述，我们的数据导致我们提出以下假设：封锁SFRP2将降低血管生成和血管肉瘤的生长，并将协同改善肿瘤回归，并结合化学疗法和抗血管生成疗法。这项研究的目的是显示这些药物在血管肉瘤中的效率，并与化学疗法和抗血管生成疗法结合使用，并阐明它们在抑制肿瘤生长中的分子机制。该项目的成功完成将导致这些药物在未来研究中的血管肉瘤患者中进行临床试验。公共卫生相关性：血管生成是新毛细血管的生长，是实体瘤生长的关键组成部分。我们发现了一种新型的蛋白质，分泌渗透的相关蛋白2，该蛋白质在人类肉瘤中过表达，它是一种潜在的血管生成刺激剂。该提案的目的是表征该蛋白在血管肉瘤生物学上的分子和细胞机制，并评估该信号途径对血管肉瘤造影术增长的oopakade有效性。