Extracellular Vesicle Proteomic Fingerprinting of Ovarian Cancer for Early Detection with a Nanoengineered Microsystem

卵巢癌细胞外囊泡蛋白质组指纹图谱用于纳米工程微系统的早期检测

• 批准号: 10621734
• 负责人: ANDREW K. GODWIN
• 金额: $ 59.3万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621734
• 关键词: 3-Dimensional; Advanced Malignant Neoplasm; Advisory Committees; Area; BRCA mutations; Benign; Biological Assay; Biological Markers; Biology; Biopsy; Blood; Blood capillaries; Blood specimen; Body Fluids; CA-125 Antigen; Cancer Control; Cancer Detection; Cancer Diagnostics; Cancer Patient; Cancer cell line; Carcinoma in Situ; Cell Line; Circulation; Clinical; Clinical Sensitivity; Colon Carcinoma; Communication; Companions; Detection; Development; Diagnosis; Diagnostic Sensitivity; Disease; Drops; Early Diagnosis; Engineering; Epithelial ovarian cancer; Excision; Fingerprint; General Population; Generations; Goals; Human; Immune response; Immunoassay; Immunoprecipitation; Intervention; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of ovary; Mammalian Oviducts; Mass Spectrum Analysis; Messenger RNA; Methods; MicroRNAs; Microfluidics; Nanochip Analytical Device; Neoplasm Circulating Cells; Operative Surgical Procedures; Outcome; Ovarian; Ovary; Pathway interactions; Patients; Pattern; Plasma; Play; Population Control; Printing; Proteins; Proteomics; Public Health; Reporting; Reproducibility; Risk; Role; Sampling; Screening for Ovarian Cancer; Screening for cancer; Sensitivity and Specificity; Serous; Serum Markers; Signaling Protein; Solid Neoplasm; Source; Specificity; Specimen; Symptoms; Technology; Testing; Time; Tissues; Training; Tumor Antigens; Tumor Markers; Tumor stage; Tumor-Derived; Ultrasonography; Untranslated RNA; Vagina; Validation; Vesicle; Woman; biomarker panel; cancer diagnosis; candidate marker; candidate validation; clinical diagnosis; clinical translation; cost; curative treatments; detection assay; diagnostic platform; disorder risk; early detection biomarkers; effusion; exosome; extracellular vesicles; high risk population; imaging modality; innovation; liquid biopsy; malignant breast neoplasm; microchip; microfluidic technology; microsystems; mortality; mutation carrier; nano; nanoengineering; nanomaterials; nanopattern; neoplastic cell; novel; novel marker; patient prognosis; pre-clinical; presymptomatic testing; prevention service; protein biomarkers; protein profiling; routine screening; screening; self assembly; tool; treatment response; tumor; tumor progression

PROJECT SUMMARY Ovarian cancer is a silent killer that strikes with few, if any, symptoms. By the time a woman knows she has it, the cancer is often advanced and the outlook grim. However, if epithelial ovarian cancer is caught early the prognosis for the patient is excellent. Developing non-invasive and highly specific blood-based tests for pre- symptomatic screening and early detection of ovarian cancer is therefore crucial. This is especially essential since obtaining a biopsy is difficult, costly, and sometimes not even an option. In addition, most blood biomarkers to date lack the necessary sensitivity and specificity for early detection of this silent killer. A fundamental challenge is the extremely low concentrations of circulating biomolecules released from the developing tumors at pre-clinical stages which can be 10,000-fold lower than their clinically detectable levels. Therefore, there is a pressing need to uncover novel biomarkers, apply new strategies, and develop robust technologies to propel the advancement of cancer diagnostics, especially in a disease such as ovarian cancer. We have focused our efforts on small extracellular vesicles (sEVs), primarily small and large exosomes derived from the endolysosomal pathway, which play important roles in cellular communication, immune response, and cancer progression via transfer of a selective repertoire of biomolecules. sEVs/exosome release is significantly increased in most neoplastic cells, including ovarian cancer and occurs continuously at all stages of tumor development. Tumor- derived sEVs accumulate in human blood and malignant effusions. These vesicles carry enriched subsets of biomolecules mirroring the tumor cells of origin, such as signaling proteins, tumor antigens, and functional RNAs (mRNA and miRNAs), which offer a new strategy to surmount the challenge in reliable detection of intrinsically low-level serum markers during early malignant transformation. Thus, the constitutive release and enrichment of certain tumor markers within sEVs present distinctive opportunities for early cancer diagnosis. We hypothesize that circulating sEVs, much akin to circulating tumor cells but more robust due to their active release and incredible stability in bodily fluids, represent a greater source for the discovery of exo-biomarkers for early detection, potentially while still confined to the fallopian tube. In addition, sEVs can serve as a 'liquid biopsy’ to assess benefits and treatment responses in real time in cancer patients. Our innovative application merges EV biology with nano-material/microfluidic technology to develop an advanced microfluidic platform to capture and detect ovarian cancer-derived sEVs with high specificity and sensitivity from the circulation. This approach will integrate validated fallopian tube and ovarian cancer associated exo-protein biomarkers (SA1), which will be applied to a second-generation nano-engineered EV analysis chip (SA2). These studies will be followed by clinical validation using longitudinal samples collected from asymptomatic women who later develop epithelial ovarian cancer (SA3). Our ultimate goal is to develop a reliable blood-based assay that, when used in conjunction with current screening approaches will decrease the mortality from ovarian cancer.

项目摘要 卵巢癌是一个沉默的杀手，几乎没有任何症状。当一个女人知道她拥有它的时候， 癌症往往是晚期，前景黯淡。然而，如果上皮性卵巢癌早期发现， 患者预后良好。开发非侵入性和高度特异性的血液检测， 因此，对卵巢癌进行症状筛查和早期发现至关重要。这一点尤为重要 因为获得活组织检查是困难的、昂贵的，并且有时甚至不是选择。此外，大多数血液生物标志物 到目前为止，缺乏必要的敏感性和特异性来早期发现这种沉默的杀手。一项基本 挑战是从发展中的肿瘤释放的循环生物分子的浓度极低 在临床前阶段，其可能比其临床可检测水平低10，000倍。因此有 迫切需要发现新的生物标志物，应用新的策略，并开发强大的技术，以推动 癌症诊断的进步，特别是在卵巢癌等疾病中。我们集中精力 在小的细胞外囊泡（sEV）上，主要是来自内溶酶体的小的和大的外泌体， 在细胞通讯、免疫应答和癌症进展中发挥重要作用， 转移生物分子的选择性库。sEV/外泌体释放在大多数人中显著增加， 肿瘤细胞，包括卵巢癌，并在肿瘤发展的所有阶段连续发生。肿瘤- 衍生的sEV在人血液和恶性积液中积累。这些囊泡携带丰富的亚群 反映肿瘤细胞起源的生物分子，如信号蛋白、肿瘤抗原和功能性RNA (mRNA和miRNAs），这提供了一种新的策略，以克服在可靠的检测固有的 低水平的血清标志物在早期恶性转化。因此，组成性释放和富集 sEV中某些肿瘤标志物的减少为早期癌症诊断提供了独特的机会。我们 假设循环sEV非常类似于循环肿瘤细胞，但由于其活性释放而更稳健， 以及体液中令人难以置信的稳定性，代表了早期发现外生物标志物的更大来源。 检测，潜在地同时仍然局限于输卵管。此外，sEV可以作为“液体活检”， 评估癌症患者的真实的时间获益和治疗反应。我们的创新应用融合了EV 生物学与纳米材料/微流体技术，以开发先进的微流体平台， 以高特异性和灵敏度从循环中检测卵巢癌来源的sEV。这种方法将 整合经验证的输卵管和卵巢癌相关外蛋白生物标志物（SA 1）， 应用于第二代纳米工程EV分析芯片（SA 2）。这些研究之后， 临床验证使用纵向样本收集无症状妇女谁后来发展上皮 卵巢癌（SA 3）。我们的最终目标是开发一种可靠的基于血液的检测方法， 与当前的筛查方法结合将降低卵巢癌的死亡率。