Composing CODAs to cervical cancer screening through an integrated CRISPR and fluorescent nucleic acid approach

通过集成 CRISPR 和荧光核酸方法将 CODA 应用于宫颈癌筛查

• 批准号: 10647930
• 负责人: Cesar M Castro
• 金额: $ 57.47万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647930
• 关键词: Acetic Acids; Adopted; Africa South of the Sahara; Algorithms; Anisotropy; Automobile Driving; Benchmarking; Bioinformatics; Biological Assay; Biomedical Engineering; Budgets; CDKN2A gene; Cause of Death; Cervical; Cervical Cancer Screening; Cessation of life; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complement; Country; Cytology; Cytopathology; DNA; DNA Markers; Decentralization; Detection; Developed Countries; Diagnostic; Disease; Early identification; Eligibility Determination; Equipment; Feedback; Fluorescence Anisotropy; Fruit; General Hospitals; Genes; Ghana; Guidelines; Gynecologic Oncology; HPV-High Risk; Human; Human Papillomavirus; Human Resources; Human papilloma virus infection; Incidence; International; Intervention; Laboratories; Machine Learning; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Massachusetts; Measurement; Messenger RNA; Motion; Nobel Prize; Nucleic Acids; Oncogenic; Optics; Pap smear; Pathology; Performance; Positioning Attribute; Provider; Public Health; RNA; RNA marker; Reporter; Resource-limited setting; Resources; Risk; Sampling; Science; Site; Specificity; Specimen; System; Technical Expertise; Technology; Temperature; Testing; Time; Traction; Triage; UPK2 gene; Uganda; Universities; Viral; Visual; Woman; cancer invasiveness; clinical diagnostics; cost; detection method; empowerment; expectation; global health; high risk; innovation; interest; iterative design; low and middle-income countries; member; multidisciplinary; multimodality; news; novel; nucleic acid detection; operation; overtreatment; pandemic disease; pilot test; point of care; portability; response; screening; user-friendly

ABSTRACT Challenges. Despite being curable when caught early, cervical cancer remains the second leading cause of death in women living in sub-Saharan Africa. Even with available screening technologies (cytology, visual inspection with acetic acid (VIA), and sometimes high-risk Human Papillomavirus [hrHPV] testing), only 10-20% of eligible women in low-and-middle-income countries (LMIC) are currently screened. Barriers cited include technical expertise, laboratory capacity, cost, and access. Most developed countries have adopted PCR-based HPV screening, furthering the resource divide with LMICs, where 80% of cancer deaths occur. Innovation. In dire response to the diagnostic gaps unveiled by the pandemic, team members within our group developed and validated in human specimens a robust and frugal technology to detect nucleic acid targets. The approach, CRISPR Optical Detection of Anisotropy or CODA, combines CRISPR/Cas (a Nobel Prize- winning technology for highly precise gene editing) with fluorescence anisotropy (differential rotational motion of fluorescent molecules). Automated nucleic acid readouts are generated in < 30 minutes. We have adopted this technology for: a) the detection of DNA and RNA markers of CIN2+; and b) reliable use by clinicians and clinical lab personnel in point-of-care settings. Our plan for this proposal is to operationalize this technology for low resource settings, notably our partner sites in Uganda and Ghana, with close LMIC input. We will then leverage CODA and other parameters to create a panel of tests to optimally detect CIN2+ in a single encounter. Aim 1: Construct a robust CODA platform for comprehensive HPV screening. Aim 2: Further examine CODA performance on human specimens and refine for LMIC operations. Aim 3: Develop a novel and rapid multi-modal algorithm for screen and treat in LMICs. Impact. Given CODA's core strengths in nucleic acid analyses (e.g., DNA or RNA), we envision an approach that yields fully quantitive readouts of high-risk HPV DNA and E6, E7, and p16 mRNA. Since the CODA assay underlies all readouts driving this proposal, a high potential exists for end-user-friendly, practical, and rapid triage of high-risk cervical disease or invasive cancer. These benefits could help decentralize and harmonize screening efforts with those guidelines currently endorsed by resource-rich countries.

抽象的 挑战。尽管早期发现可以治愈，但宫颈癌仍然是导致宫颈癌的第二大原因。 生活在撒哈拉以南非洲的妇女死亡。即使有可用的筛查技术（细胞学、视觉 仅用醋酸 (VIA) 检查，有时还进行高危人乳头瘤病毒 [hrHPV] 检测） 目前，中低收入国家 (LMIC) 10-20% 的符合资格的女性接受了筛查。引用的障碍 包括技术专长、实验室能力、成本和访问权限。大多数发达国家已采用 基于 PCR 的 HPV 筛查，进一步扩大了与中低收入国家的资源差距，80% 的癌症死亡发生在这些国家。 创新。为了对这一流行病所揭示的诊断差距做出严峻的反应，我们小组内的团队成员 开发并在人体样本中验证了一种强大且节俭的技术来检测核酸目标。 该方法被称为 CRISPR 各向异性光学检测（CODA），结合了 CRISPR/Cas（诺贝尔奖 具有荧光各向异性（微分旋转运动）的高精度基因编辑获胜技术 荧光分子）。不到 30 分钟即可生成自动核酸读数。我们已经采用了 该技术用于：a) CIN2+的DNA和RNA标记物的检测； b) 临床医生的可靠使用和 护理点环境中的临床实验室人员。我们对此提案的计划是将这项技术投入使用 低资源环境，特别是我们在乌干达和加纳的合作伙伴站点，有密切的中低收入国家投入。我们随后将 利用 CODA 和其他参数创建一组测试，以在单个测试中最佳地检测 CIN2+ 遇到。目标 1：构建强大的 CODA 平台，用于全面的 HPV 筛查。目标 2：进一步 检查 CODA 在人体样本上的性能并针对 LMIC 操作进行改进。目标 3：创作一部小说 用于中低收入国家筛查和治疗的快速多模式算法。影响。鉴于CODA在核酸方面的核心优势 酸分析（例如 DNA 或 RNA），我们设想了一种能够完全定量读出高风险的方法 HPV DNA 和 E6、E7 和 p16 mRNA。由于 CODA 测定是推动该提案的所有读数的基础， 对最终用户友好、实用且快速的高危宫颈疾病或侵入性疾病进行分类的潜力巨大 癌症。这些好处可以帮助分散筛查工作并使其与当前的指南相协调 受到资源丰富国家的认可。