Development of An Innovative TEE Technology for Mutation Detection

开发用于突变检测的创新 TEE 技术

基本信息

批准号：
10757697
负责人：
Qipan Deng
金额：
$ 107.16万
依托单位：
GLC BIOTECHNOLOGY, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-18 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10757697
关键词：
Aftercare Alleles BRAF gene Biological Models Blood Cancer Detection Cancer Patient Clinical Codon Nucleotides DNA DNA Mutational Analysis DNA Sequence Detection Development Diagnostic tests Dideoxy Chain Termination DNA Sequencing Drug Targeting Effectiveness Eligibility Determination Epidermal Growth Factor Receptor FDA approved Foundations Future KRAS2 gene Malignant Neoplasms Managed Care Medicine Methods Monitor Mutate Mutation Mutation Analysis Mutation Detection Patients Performance Pharmaceutical Preparations Phase Plasma Recovery Reproducibility Sampling Serum Small Business Innovation Research Grant Specificity Specimen System Systems Analysis Technology Testing Tissue Sample Tissues Tumor Tissue Validation Work cancer care clinical application commercialization companion diagnostics cost detection test digital improved innovation liquid biopsy novel success targeted treatment technology platform tumor validation studies

项目摘要

Abstract The objective of this project is to further develop a breakthrough Toehold-Enrichment- Extraction (TEE) technology and validate TEE-based tests for use in cancer detection and care management. TEE is a novel DNA extraction method. Unlike conventional extraction methods, TEE extracts mutated DNA while enriching it with high recovery. Moreover, unlike current PCR- based enrichment methods, TEE enriches mutated DNA without altering original DNA sequences. We have shown that TEE could enrich mutated DNA by as much as 1000 fold. Therefore, when TEE is used to extract DNA for a mutation detection test, it significantly increases analytical sensitivity of the test, leading to accurate detection of mutated DNA at a much lower concentration level, which could not be achieved if a conventional method is used to extract DNA. To illustrate, when a conventional method is used to extract DNA, Sanger sequencing cannot detect mutated DNA if its concentration is less than 20%. In contrast, when TEE is used to extract mutated DNA, it enables Sanger sequencing to detect 0.1% mutated DNA. In other words, Sanger-sequencing becomes as sensitive as digital PCR or NGS when we incorporate TEE into Sanger sequencing (we term this combination TEE-Sanger sequencing) for mutation analysis. Furthermore, we have demonstrated that TEE-Sanger sequencing could detect mutated DNA from clinical specimen like FFPE tumor tissue and blood (serum) samples, indicating that TEE-based testing is compatible with clinical applications. Since TEE is a DNA extraction method, it can replace conventional methods to extract DNA for mutation analysis. Moreover, TEE is not only a breakthrough technology, but also a platform on which various TEE-based tests can be developed for different clinical applications. Inspired by the success of our preliminary study and potential of the TEE technology, we propose this SBIR Phase II project to further study the TEE technology, providing a strong foundation for its commercialization. This project has four specific aims.

抽象的该项目的目标是进一步开发突破性的立足点-丰富- 提取 (TEE) 技术并验证基于 TEE 的测试在癌症检测和护理中的使用管理。 TEE是一种新颖的DNA提取方法。与传统的提取方法不同， TEE 提取突变 DNA，同时以高回收率富集它。此外，与目前的PCR不同的是基于富集方法，TEE 富集突变 DNA，而不改变原始 DNA 序列。我们已经证明 TEE 可以将突变 DNA 富集多达 1000 倍。因此，当 TEE用于提取DNA进行突变检测测试，它显着提高了分析能力测试灵敏度高，可在低得多的浓度下准确检测突变 DNA 这是用传统方法提取DNA无法达到的水平。为了说明这一点，当使用常规方法提取DNA时，桑格测序无法检测到突变 DNA，如果其浓度低于 20%。相比之下，当使用TEE提取突变DNA时，它使桑格测序能够检测 0.1% 的突变 DNA。换句话说，桑格测序当我们将 TEE 纳入桑格测序时，变得与数字 PCR 或 NGS 一样灵敏（我们将这种组合称为 TEE-Sanger 测序）用于突变分析。此外，我们还有证明 TEE-Sanger 测序可以检测临床样本中的突变 DNA，例如 FFPE肿瘤组织和血液（血清）样本，表明基于TEE的检测是兼容的与临床应用。由于TEE是一种DNA提取方法，因此可以替代传统的提取方法用于突变分析的 DNA。而且，TEE不仅是一项突破性的技术，更是一种平台上可以针对不同的临床应用开发各种基于 TEE 的测试。受到我们初步研究的成功和 TEE 技术潜力的启发，我们建议此次SBIR二期项目进一步研究TEE技术，为它的商业化。该项目有四个具体目标。