Development of aptamer-based detection and therapy strategies for ovarian cancer

开发基于适体的卵巢癌检测和治疗策略

• 批准号: 9433931
• 负责人: Rebecca Jean Whelan
• 金额: $ 3.7万
• 依托单位: OBERLIN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9433931
• 关键词: Adverse effects; Affinity; Antibodies; Aptamer Technology; Back; Binding; Biological; Biological Assay; Biological Markers; Blood; CA-125 Antigen; Capillary Electrophoresis; Cell surface; Cells; Cessation of life; Complement; Detection; Development; Diagnosis; Disease; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Epididymis; Fluorescence Polarization; General Population; Goals; Gold; Health Benefit; Home environment; Human; Hyperthermia; Image; Immobilization; In Vitro; Institution; Label; Ligands; Localized Malignant Neoplasm; Malignant neoplasm of ovary; Measures; Mediating; Mentors; Metals; Methods; Nucleic Acids; Ovary; Patients; Pharmaceutical Preparations; Phototherapy; Physicians; Process; Proteins; Public Health; Research; Research Project Grants; Sampling; Scanning Electron Microscopy; Screening for Ovarian Cancer; Screening for cancer; Sensitivity and Specificity; Serum; Single-Stranded DNA; Spatial Distribution; Specificity; Spectrum Analysis; Surface; Surface Plasmon Resonance; Survival Rate; Techniques; Temperature; Testing; Therapeutic; Time; Training; Translating; Tumor Markers; United States; Universities; Wisconsin; Woman; advanced disease; analytical method; aptamer; assay development; base; cancer biomarkers; cancer cell; cancer diagnosis; cancer therapy; cell killing; cellular imaging; chemotherapy; college; diagnostic assay; experimental study; high resolution imaging; imaging agent; improved; improved outcome; infancy; innovation; insight; interest; killings; medical schools; mesothelin; mortality; nanomaterials; nanoparticle; neoplastic cell; novel; ovarian neoplasm; protein biomarkers; resilience; sabbatical; screening; statistics; tumor; undergraduate student

DESCRIPTION (provided by applicant): The need persists for better early detection of ovarian cancer. Currently, detection of ovarian cancer relies in part on the recognition of the tumor marker protein CA125 by antibody molecules. CA125 is the most widely assayed and best validated biomarker of ovarian cancer, but because available assays lack sufficient sensitivity and specificity, CA125 is not currently recommended for screening in the general population. More informative assays will result from the simultaneous detection of multiple cancer biomarkers, including those whose biological abundance complements CA125. Mesothelin and human epididymis protein 4 (HE4) are two relatively new biomarkers that may complement CA125 in serum tests. Currently, most assays for ovarian cancer markers rely on antibody molecules for selective recognition. Antibodies are limited in terms of their stability, reusability, and ability to be modified with imaging labels or immobilization tags. Assays incorporating aptamers (functional nucleic acid ligands) are an attractive alternative to antibody-based techniques. Aptamers are robust, easily labeled, and prepared entirely in vitro. In previous studies, some aptamers have been shown surpass antibodies in affinity for their target, which may translate into improved detection methods for cancer markers in serum tests. An additional application in which aptamers may replace antibodies is in targeting metal nanoparticle imaging agents or therapeutic payloads directly to surfaces of ovarian cancer cells. Such cell targeting may be useful for imaging the distribution of biomarkers on ovarian tumors and may enable localized cell killing by hyperthermia, reducing the need for systemic chemotherapy. The research plan proposed here aims to develop aptamers for three important ovarian cancer biomarkers. Assays that use these aptamers in conjunction with sensitive instrumental methods will then be developed. We will also perform proof-of-principle experiments in which aptamer-functionalized nanoparticles mediate high-resolution imaging and targeted hyperthermia. Public health will be positively impacted by the development of new CA125 assays to improve cancer screening and by innovative approaches to tumor cell eradication. The PI will conduct the metal nanoparticle assays during a one-year research sabbatical hosted at the University of Wisconsin Medical School (Year 1). Two additional years of support are requested during which the PI, back at her home institution of Oberlin College, will train two undergraduate students per year and mentor them in aptamer selection and assay development. A recently graduated Oberlin undergraduate will be employed as a full-time research technician in Year 3.

描述（由申请人提供）：对于更好地早期检测卵巢癌的需求仍然存在。目前，卵巢癌的检测部分依赖于抗体分子对肿瘤标志蛋白CA125的识别。 CA125 是检测最广泛、验证最好的卵巢癌生物标志物，但由于现有检测缺乏足够的敏感性和特异性，目前不建议在一般人群中使用 CA125 进行筛查。同时检测多种癌症生物标志物，包括那些生物丰度与 CA125 互补的生物标志物，将产生信息更丰富的检测结果。间皮素和人附睾蛋白 4 (HE4) 是两种相对较新的生物标志物，可以在血清测试中补充 CA125。目前，大多数卵巢癌标记物的检测依赖于抗体分子进行选择性识别。抗体的稳定性、可重复使用性以及使用成像标签或固定标签进行修饰的能力受到限制。结合适体（功能性核酸配体）的检测是基于抗体的技术的一种有吸引力的替代方案。适体坚固耐用，易于标记，并且完全在体外制备。在之前的研究中，一些适配体已被证明对其靶点的亲和力超过了抗体，这可能会转化为血清测试中癌症标志物检测方法的改进。适体可以替代抗体的另一个应用是将金属纳米颗粒成像剂或治疗有效负载直接靶向卵巢癌细胞的表面。这种细胞靶向可能有助于对卵巢肿瘤上生物标志物的分布进行成像，并且可以通过热疗实现局部细胞杀伤，从而减少全身化疗的需要。这里提出的研究计划旨在开发三种重要卵巢癌生物标志物的适体。然后将开发使用这些适体与敏感仪器方法结合的测定方法。我们还将进行原理验证实验，其中适配体功能化纳米粒子介导高分辨率成像和靶向热疗。改进癌症筛查的新 CA125 检测方法的开发以及肿瘤细胞根除的创新方法将对公共健康产生积极影响。 PI 将在威斯康星大学医学院（第一年）举办的为期一年的研究休假期间进行金属纳米颗粒测定。需要额外两年的支持，在此期间，PI 将回到她所在的欧柏林学院，每年培训两名本科生，并指导他们进行适体选择和检测开发。刚毕业的欧柏林本科生将在第三年受聘担任全职研究技术员。

项目成果

Selection of DNA Aptamers for Ovarian Cancer Biomarker CA125 Using One-Pot SELEX and High-Throughput Sequencing.

使用一锅 SELEX 和高通量测序选择卵巢癌生物标志物 CA125 的 DNA 适体。

• 发表时间: 2017
• 作者: Scoville, Delia J;Uhm, Tae Kyu Brian;Shallcross, Jamie A;Whelan, Rebecca J
• 通讯作者: Whelan, Rebecca J

Effects of Cationic Proteins on Gold Nanoparticle/Aptamer Assays.

阳离子蛋白对金纳米粒子/适体测定的影响。

• 发表时间: 2017-11-30
• 影响因子: 4.1
• 作者: Pires, Thomas A;Narovec, Conor M;Whelan, Rebecca J
• 通讯作者: Whelan, Rebecca J

Selection of DNA aptamers for ovarian cancer biomarker HE4 using CE-SELEX and high-throughput sequencing.

使用 CE-SELEX 和高通量测序选择卵巢癌生物标志物 HE4 的 DNA 适体。

• 发表时间: 2015-09
• 影响因子: 4.3
• 作者: Eaton, Rachel M;Shallcross, Jamie A;Mael, Liora E;Mears, Kepler S;Minkoff, Lisa;Scoville, Delia J;Whelan, Rebecca J
• 通讯作者: Whelan, Rebecca J

Experimental and mathematical evidence that thrombin-binding aptamers form a 1 aptamer:2 protein complex.

实验和数学证据表明凝血酶结合适体形成 1 适体：2 蛋白质复合物。

• 发表时间: 2018
• 作者: Mears, Kepler S;Markus, Daniel L;Ogunjimi, Oluwadamilare;Whelan, Rebecca J
• 通讯作者: Whelan, Rebecca J