Treatment of breast cancer tumor growth and metastasis with an anti-MMP-9 DNAzyme

用抗 MMP-9 DNAzyme 治疗乳腺癌肿瘤生长和转移

基本信息

批准号：
8225421
负责人：
Miranda Alvina Marie Hallett
金额：
$ 1.68万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-12-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8225421
关键词：
Achievement Affect Animal Model Animals Basement membrane Binding Biological Assay Biological Sciences Brain Breast Cancer Treatment Breast Carcinoma Cancer Patient Catalytic DNA Cause of Death Cell Proliferation Cell Transplants Cleaved cell Cytokine Degradation Development Diagnosis Diffusion Dose Down-Regulation Early Diagnosis Enzymes Epithelial Cells Extracellular Matrix Extravasation Fatty acid glycerol esters Female Gelatinase B Goals Heart Image In Vitro Injection of therapeutic agent Kidney Label Lead Life Liver Luc Gene Luciferases Lung Lung nodule Malignant Epithelial Cell Malignant Neoplasms Mammary Neoplasms Mammary gland Mastectomy Matrix Metalloproteinases Measurement Measures Mediating Messenger RNA Metabolic Clearance Rate Metalloproteases Microscopy Monitor Morbidity - disease rate Mouse Mammary Tumor Virus Mus Neoplasm Metastasis Neoplasm Transplantation Oligonucleotides Operative Surgical Procedures Organ Palpable Patients Play Proteins Quality of life Reporting Research Role Series Slice Survival Rate System Tail Techniques Time Tissues Toxic effect Transgenic Mice Transplantation Tumor Burden Tumor Cell Invasion Tumor Tissue Veins Woman angiogenesis bone cell stroma chemotherapy cytotoxicity efficacy testing in vivo innovation lung development lymph nodes malignant breast neoplasm mortality mouse model neoplastic cell novel outcome forecast prevent protein expression research study tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Despite continued improvements in diagnosis, surgical techniques, and chemotherapy, breast cancer patients are still being overcome by cancer metastasis. Breast cancer metastasizes to brain, lung, bone, liver and regional lymph nodes. Early detection and advancements in research have increased survival and enhanced quality of life. However, there Is still a need for improvement. Tumor cell proliferation, invasion, and metastasis are known to be mediated, at least in part, through degradation of basement membrane by neutral metalloproteinases (MMP) produced by tumor and stroma cells. Evidence suggests that MMP-9 plays a significant role in breast tumor cell invasion and metastasis. We have shown that DNAzyme generated against MMP-9 mRNA was able to reduce the expression of the enzyme in vitro and its intratumoral injection reduced the size of mammary tumor, generated in a MMTV- PyMT transgenic mouse model, by 50%. DNAzymes are catalytic DNA molecules that specifically bind and cleave mRNA of the targeted protein, resulting in decreased protein expression. Given their potential for systemic administration and targeting, anti-MMP-9 DNAzyme (AM9D) could prove useful as a therapy against breast carcinoma invasion. We hypothesize the down regulation of MMP-9, by AM9D, will lead to the inhibition of tumor growth, decrease in lung metastasis, block experimental metastasis and increase survival in breast carcinoma animal model. Towards the achievement of this goal we propose to (1) determine the minimum effective dose of AM9D needed to significantly decrease tumor load in the PyMT MTEC transplant mouse model without causing significant local toxicity. Toxicity will be determined by histopathological analysis of tissue slices. (2) assess the affect of AM9D on development of secondary tumor. Luciferase expressing PyMT MTECs will be transplanted into female mice mammary glands of female mice and tumors will be treated with an optimum concentration of AM9D, control DNAzyme or PBS for 4 weeks. Tumors will then be removed and animal will be maintained for an additional 8 weeks to develop lung secondary tumors. In addition, experimental metastasis will be performed by injecting PyMT MTEC with or without AM9D or control DNazyme into tail vein of FVB/Nj mice. Formation and development of lung metastasis will then be followed by in vivo Xenogen bioimager system. (3) The effect of DNAzyme treatment on the survival rate of animals bearing breast carcinomas will be assessed. Thus, in this proposal we are taking an innovative approach to the treatment of breast carcinomas by studying the effect of the AM9D therapy on tumor growth and metastasis, the major mechanism of morbidity and mortality.

描述（由申请人提供）：尽管诊断、手术技术和化疗不断改进，但乳腺癌患者仍然受到癌症转移的困扰。乳腺癌转移至脑、肺、骨、肝和区域淋巴结。早期发现和研究进展提高了生存率并提高了生活质量。然而，仍然需要改进。已知肿瘤细胞增殖、侵袭和转移至少部分是通过肿瘤和基质细胞产生的中性金属蛋白酶（MMP）降解基底膜来介导的。有证据表明MMP-9在乳腺肿瘤细胞侵袭和转移中发挥重要作用。我们已经证明，针对 MMP-9 mRNA 生成的 DNAzyme 能够在体外降低该酶的表达，并且其瘤内注射可将 MMTV-PyMT 转基因小鼠模型中产生的乳腺肿瘤的大小减小 50%。 DNAzyme 是催化 DNA 分子，可特异性结合并切割目标蛋白的 mRNA，从而导致蛋白表达降低。鉴于其全身给药和靶向的潜力，抗 MMP-9 DNAzyme (AM9D) 可能被证明可用作对抗乳腺癌侵袭的疗法。我们假设 AM9D 对 MMP-9 的下调将导致乳腺癌动物模型中肿瘤生长的抑制、肺转移的减少、实验性转移的阻断并增加存活率。为了实现这一目标，我们建议 (1) 确定显着降低 PyMT MTEC 移植小鼠模型中肿瘤负荷而不引起显着局部毒性所需的 AM9D 最小有效剂量。毒性将通过组织切片的组织病理学分析来确定。 (2)评估AM9D对继发性肿瘤发生的影响。将表达荧光素酶的 PyMT MTEC 移植到雌性小鼠的乳腺中，并用最佳浓度的 AM9D、对照 DNAzyme 或 PBS 处理肿瘤 4 周。然后将切除肿瘤并将动物再饲养8周以形成肺部继发性肿瘤。此外，通过将含有或不含AM9D或对照DNAzyme的PyMT MTEC注射到FVB/Nj小鼠的尾静脉中来进行实验转移。体内 Xenogen 生物成像系统将跟踪肺转移的形成和发展。 (3)将评估DNAzyme治疗对患有乳腺癌的动物的存活率的影响。因此，在本提案中，我们通过研究 AM9D 疗法对肿瘤生长和转移（发病率和死亡率的主要机制）的影响，采取创新的方法来治疗乳腺癌。