CPE Peptide-Based Nanoparticles for the Diagnosis and Therapy of Chemotherapy Res

基于CPE肽的纳米粒子用于化疗的诊断和治疗研究

• 批准号: 8165297
• 负责人: Alessandro D Santin
• 金额: $ 34.34万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8165297
• 关键词: Affinity; Amino Acids; Animal Model; Animals; Binding; Breast; Cancer Patient; Cell Line; Cells; Clinical Trials; Clostridium perfringens enterotoxin; Clostridium perfringens enterotoxin receptor; Complex; Coumarins; Cytolysis; Cytotoxic agent; Data; Development; Diagnosis; Disease; Disease Resistance; Doxorubicin; Drug Kinetics; Encapsulated; Epithelial; Epithelial Cells; Fingerprint; Fluorescein; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Foundations; Gene Expression; Genes; Glycolates; Goals; Gynecologic; In Vitro; Label; Length; Local Therapy; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Membrane; Mesothelial Cell; Modeling; Molecular Profiling; Molecular Target; Mus; Normal Cell; Oligonucleotide Microarrays; Operative Surgical Procedures; Organ; Ovarian; Ovarian Carcinoma; Ovarian Diseases; Pancreas; Papillary Neoplasm; Patients; Peptide Fragments; Peptides; Pharmaceutical Preparations; Positioning Attribute; Prostate; Proteins; Public Health; Receptor Cell; Recurrence; Research; Resistance; Serous; Suicide; Testing; Therapeutic; Therapeutic Agents; Tight Junctions; Toxin; Translating; Treatment Efficacy; WFDC2 gene; Xenograft procedure; alpha benzopyrone; base; cancer cell; chemotherapy; claudin 3; claudin 4; clinically relevant; design; domain mapping; effective therapy; genetic analysis; high throughput technology; human tissue; improved; in vitro Assay; in vivo; innovation; mesothelin; mortality; nanoparticle; neoplastic cell; novel; ovarian neoplasm; overexpression; plasmid DNA; preclinical study; prevent; promoter; receptor; research study; therapeutic target; tumor

DESCRIPTION (provided by applicant): Ovarian carcinoma remains the cancer with the highest mortality rate among gynecological tumors. Although many ovarian cancer patients fully respond to the standard combination of surgery and chemotherapy, nearly 90% later develop recurrent chemotherapy-resistant cancer and inevitably succumb to their disease. Thus, development of innovative, effective therapies against recurrent/chemotherapy-resistant ovarian cancer remains a high priority for improving public health. Using high-throughput technologies to analyze genetic fingerprints of ovarian cancer, we have recently discovered extremely high expression of the genes encoding the proteins claudin-3 and claudin-4. Because claudin-3 and -4 are the epithelial receptors for Clostridium perfringens enterotoxin (CPE), and are sufficient to mediate CPE binding, we hypothesize that using biodegradable nanoparticles such as poly (lactic-co-glycolic acid, i.e., PLGA-NP) encapsulating therapeutic agents complexed to the binding domain of CPE, to target ovarian cancer cells, is a novel, potentially highly effective therapeutic approach to treat chemotherapy-resistant ovarian cancer. Consistent with this view, preliminary in vitro data clearly showed that fluorescein (FITC)-labeled-C-CPE effectively binds and rapidly internalizes in chemotherapy-resistant primary OSPC cell lines, suggesting that C- CPE is capable to bind and also of translocating drugs across cell tumor membrane. More importantly, preliminary in vivo results showed that CPE-PLGA-NP encapsulating a fluorescent dye (coumarin) selectively accumulate in vivo in chemotherapy resistant ovarian tumors with minimal nonspecific accumulation in RES organs and only background binding in normal cells of various other organs. Accordingly, this proposal has three related specific aims: 1) Characterize fluorescent PLGA-NP encapsulating model and therapeutic drugs, complexed to CPE peptides of different lengths (i.e., 30aa, 17aa and 9 aa), and evaluate binding activity and therapeutic efficacy of such NP in multiple in vitro assays against primary chemotherapy resistant ovarian carcinoma cell lines, 2) Examine the distribution and pharmacokinetics of PLGA-NP encapsulating coumarin complexed to 125I labeled C-CPE peptide in vivo in clinically relevant animal models of primary chemotherapy resistant ovarian carcinoma and 3) Examine the therapeutic potential of PLGA-NP encapsulating plasmid DNA encoding Diptheria Toxin A suicide protein under the control of promoter sequences of genes highly and preferentially active in ovarian cancer (i.e., mesothelin and HE4/WFDC2), complexed to CPE peptide in vivo in clinically relevant animal models of primary chemotherapy resistant ovarian carcinoma. Upon completion of this project, we will be positioned to quickly translate this research into a novel, highly effective therapeutic treatment for patients with chemotherapy-resistant disease. PUBLIC HEALTH RELEVANCE: Our research group has recently discovered extremely high expression of the genes encoding the proteins claudin-3 and claudin-4 in ovarian cancer, the most lethal gynecologic malignancy. Because claudin-3 and -4 are the epithelial receptors for Clostridium perfringens enterotoxin (CPE), and are sufficient to mediate CPE binding, in the proposed research we plan to evaluate the use of biodegradable polymeric nanoparticles such as poly(lactic-co-glycolic acid, i.e., PLGA NP) encapsulating plasmid DNA encoding Diptheria Toxin A suicide protein under the control of promoter sequences of genes highly and preferentially active in ovarian cancer (i.e., mesothelin and HE4/WFDC2), complexed to the binding domain of CPE, as a novel, potentially highly effective therapeutic approach to treat chemotherapy-resistant ovarian cancer. Upon completion of this project, we will be positioned to quickly translate this research into an innovative and highly targeted therapeutic treatment for patients with chemotherapy-resistant disease.

描述（申请人提供）：卵巢癌仍然是妇科肿瘤中死亡率最高的癌症。尽管许多卵巢癌患者对手术和化疗的标准组合完全有效，但近90%的患者后来发展为复发性化疗耐药癌症，并不可避免地死于疾病。因此，开发针对复发性/化疗耐药卵巢癌的创新、有效疗法仍然是改善公众健康的高度优先事项。利用高通量技术分析卵巢癌的遗传指纹，我们最近发现了编码蛋白质claudin-3和claudin-4的基因的极高表达。由于claudio -3和-4是产气荚膜梭菌肠毒素（CPE）的上皮受体，并且足以介导CPE的结合，我们假设使用可生物降解的纳米颗粒，如聚乳酸-羟基乙酸，即PLGA-NP，包封治疗药物与CPE的结合域，靶向卵巢癌细胞，是一种新的，潜在的高效治疗方法，用于治疗化疗耐药的卵巢癌。与这一观点一致的是，初步的体外实验数据清楚地表明，荧光素（FITC）标记的C-CPE在化疗耐药的原代OSPC细胞系中有效结合并迅速内化，这表明C-CPE能够结合并跨肿瘤细胞膜转运药物。更重要的是，初步的体内实验结果表明，CPE-PLGA-NP包被一种荧光染料（香豆素）在体内选择性地积累在耐化疗卵巢肿瘤中，在RES器官中具有最小的非特异性积累，仅在其他器官的正常细胞中具有背景结合。因此，这项建议有三个相关的具体目标：1)对不同长度CPE肽（30aa、17aa和9aa）的PLGA-NP荧光包封模型和治疗药物进行表征，并通过多次体外实验评价该NP对原发性化疗耐药卵巢癌细胞株的结合活性和治疗效果；2)在原发性化疗耐药卵巢癌临床相关动物模型中，检测PLGA-NP包封香豆素与125I标记的C-CPE肽复合物在体内的分布和药代动力学；3)在卵巢癌高优先活性基因启动子序列（即间皮素和HE4/WFDC2）的控制下，检测PLGA-NP包封编码白喉毒素A自杀蛋白的质粒DNA的治疗潜力。在临床相关的原发性化疗耐药卵巢癌动物模型中与CPE肽络合。在这个项目完成后，我们将定位于快速将这项研究转化为一种新的，高效的治疗化疗耐药疾病的治疗方法。