A Novel Method for Efficiently and Robustly Retrieving Circulating miRNAs

一种高效、稳健地检索循环 miRNA 的新方法

• 批准号: 10013246
• 负责人: Qipan Deng
• 金额: $ 75万
• 依托单位: GLC BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-09 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013246
• 关键词: Adsorption; Biological Assay; Biological Markers; Biological Models; Blood; Cardiovascular Diseases; Clinical; DNA; Detection; Development; Diagnosis; Disease; FDA approved; Foundations; Health; Human; Individual; Interruption; Malignant Neoplasms; Methods; MicroRNAs; Monitor; Nucleic Acids; Phase; Plasma; Protocols documentation; RNA; Sampling; Serum; Small Business Innovation Research Grant; Specimen; Surface; Technology; Testing; base; cancer biomarkers; circulating microRNA; clinical application; colorectal cancer screening; commercialization; cost; cost effective; disease diagnosis; improved; microRNA biomarkers; new technology; novel; success

Abstract This direct SBIR Phase II project aims to develop a novel technology and associated assays for efficiently and robustly extracting circulating miRNAs from blood (i.e. plasma or serum), which could transform the utility of miRNA testing in the diagnosis and monitoring of major diseases such as cancer and cardiovascular disease. Our competitive advantage lies in the ability to retrieve circulating miRNAs more efficiently and robustly than current extraction methods, which has been a critical barrier in implementing miRNA testing clinically. This advantage is achieved by implementing sequence specific capture in conjunction with a propriety method of making novel capture beads. Circulating miRNAs are potential disease biomarkers. However, to date, there are no FDA approved miRNA tests available. A bottleneck problem is unreliable circulating miRNA extraction. Current extraction methods are based on adsorption of polar molecules on polar surfaces, which were originally developed to extract large DNA and RNA molecules. However, because miRNAs are so small (~22nt), their interaction with polar surfaces is much weaker, therefore their adsorption on polar surfaces can be easily interrupted by other molecules present in sample. Since the weak adsorption of miRNAs on polar surfaces is an inherent problem that cannot be fully solved even if all other conditions were optimized, new methods for retrieving circulating miRNAs based on a different mechanism are clearly needed. Sequence-specific capture (SSC) is another method to extract nucleic acids, but historically it performs poorly when used to extract circulating nucleic acids from clinical samples due to the lack of effective capture beads. In addition, the cost of making capture beads by current methods is very high, making SSC too expensive for clinical use. Recently, we made a major breakthrough by developing a proprietary method of making capture beads that can transform SSC from a concept to a practical method for extracting circulating miRNAs in an efficient, robust, and cost-effective manner. Therefore, we employed our capture beads to develop our own SSC assays, and conducted a systematic study to examine the feasibility of using our SSC assay in extracting circulating miRNAs. Our SSC assay was indeed found to efficiently and robustly extract circulating miRNAs. In the study, we also found that extraction efficiency of current kits varied by as much as 60-fold by plasma sample, further confirming that current methods are not robust. Considering the problem of current methods, unparalleled features of our SSC technology, and success of our early study, we propose this Phase II SBIR project to further develop and validate our SSC technology for extracting circulating miRNAs from plasma/serum, providing a foundation for its commercialization.

抽象的 这个直接的SBIR II期项目旨在开发一种新颖的技术和相关的测定，以有效和 从血液（即血浆或血清）中鲁棒提取循环miRNA，这可能会改变 在诊断和监测主要疾病（例如癌症和心血管疾病）中的miRNA测试。 我们的竞争优势在于能够更有效，更强大地检索循环miRNA 当前的提取方法，这是在临床上实施miRNA测试的关键障碍。这 通过与专有方法结合实现序列特定捕获来实现优势 制作新颖的捕获珠。 循环miRNA是潜在的疾病生物标志物。但是，迄今为止，还没有FDA批准的miRNA 可用测试。瓶颈问题是不可靠的循环miRNA提取。当前提取方法 基于极性分子在极面上的吸附，这些分子最初是为了提取大型而开发的 DNA和RNA分子。但是，由于miRNA是如此小（〜22nt），因此它们与极性的相互作用 表面要弱得多，因此它们在极地表面上的吸附很容易被其他 样品中存在的分子。由于miRNA在极地表面上的吸附弱是一个固有的问题 即使所有其他条件都已优化，这也无法完全解决，这是检索循环的新方法 显然需要基于不同机制的miRNA。序列特异性捕获（SSC）是另一个 提取核酸的方法，但从历史上看，它用来提取循环核的方法很差 由于缺乏有效的捕获珠，来自临床样品的酸。另外，捕获的成本 当前方法的珠子非常高，因此SSC对于临床使用而言太昂贵了。 最近，我们通过开发一种专有的方法来制作捕获珠，取得了重大突破 可以将SSC从概念转变为一种实用方法，用于在有效，健壮， 和成本效益的方式。因此，我们采用捕获珠来开发自己的SSC分析，并 进行了一项系统的研究，以检查使用我们的SSC分析提取循环的可行性 mirnas。确实发现我们的SSC分析可以有效且可靠地提取循环miRNA。在研究中， 我们还发现，当前试剂盒的提取效率通过等离子体样本而变化多达60倍 确认当前方法不强大。 考虑当前方法的问题，我们的SSC技术的无与伦比的特征以及我们的成功 早期研究，我们建议该II阶段SBIR项目，以进一步开发和验证我们的SSC技术 从血浆/血清中提取循环miRNA，为其商业化提供了基础。