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：
8472337
负责人：
Nancy Demore
金额：
$ 28万
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/8472337
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侵袭性血管恶性肿瘤，具有高转移潜力和随后的死亡率(1) 诊断为血管肉瘤时，10¿ 25%的患者已经有所有血管肉瘤患者的 2 年和 5 年总生存率为 50% 和 30%，对于不可切除的血管肉瘤，基于阿霉素的中位总生存期分别为 24 个月。治疗方案的无进展生存期为 5 个月，紫杉醇的无进展生存期为 7 个月因此，迫切需要新的疗法来提高患者的生存率。我们的实验室最近发现了一种导致高度致命疾病的新信号通路。与正常血管细胞相比，对乳腺肿瘤血管细胞进行基因组分析。通过激光捕获显微切割获得的血管，我们鉴定出分泌性卷曲相关蛋白 2 (SFRP2) 肿瘤内皮中表达增加的基因 SFRP2 是一种参与 Wnt 的 33kd 分泌蛋白。通过 IHC，我们的初步数据显示 SFRP2 蛋白存在于 9/9 的人类血管肉瘤中，并且刺激鸡绒毛尿囊膜上的血管生成，诱导内皮细胞迁移和管化 SFRP2 的形成，并防止缺氧诱导的细胞凋亡。导致管形成受到抑制，用 SFRP2 刺激内皮细胞导致管形成增加 NFATc3 是一种转录因子，在介导血管生成中发挥着关键作用。 NFAT 核定位取决于动态的进出口平衡。 Ca2+/钙调蛋白依赖性磷酸酶、钙调神经磷酸酶的活性以及丝氨酸/苏氨酸的活性 NFAT 在去磷酸化之前不能正常进入细胞核，但可以被激活。激活的钙调神经磷酸酶使 NFAT 去磷酸化，然后从细胞质转移到细胞质。细胞核并导致参与细胞生长、分化和细胞周期进展的基因转录。他克莫司 (FK506) 是一种免疫抑制药物，可与淋巴细胞中的亲免蛋白 FKBP12 结合。 FK506-FKBP12 复合物与钙调神经磷酸酶结合并抑制其磷酸酶活性，从而抑制 NFAT(5) 的核转位我们的初步数据表明他克莫司抑制血管肉瘤管。体外形成，全身给药抑制裸鼠SVR血管肉瘤异种移植物的生长总而言之，我们的数据使我们在 20 天时减少了 46%。 SFRP2 将减少血管生成和血管肉瘤生长，并协同改善肿瘤消退本研究的目的是展示与化疗和抗血管生成疗法相结合。这些药物单独治疗血管肉瘤以及与化疗和抗血管生成药物联合治疗的疗效治疗，并阐明其抑制肿瘤生长的分子机制。该项目将在未来的研究中对这些药物在血管肉瘤患者中进行临床试验。