用于癌症检测的单个小细胞外囊泡的表面等离子体纳米孔无标记成像

Plasmonic nano-aperture label-free imaging of single small extracellular vesicles for cancer detection.

作者信息

Ohannesian Nareg, Mallick Mohammad Sadman, He Jianzhong, Qiao Yawei, Li Nan, Shaitelman Simona F, Tang Chad, Shinn Eileen H, Hofstetter Wayne L, Goltsov Alexei, Hassan Manal M, Hunt Kelly K, Lin Steven H, Shih Wei-Chuan

机构信息

Department of Electrical and Computer Engineering, University of Houston, 4800 Martin Luther King Blvd., Houston, TX, 77204, USA.

Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd, Houston, TX, 77030, USA.

出版信息

Commun Med (Lond). 2024 May 25;4(1):100. doi: 10.1038/s43856-024-00514-x.

DOI:10.1038/s43856-024-00514-x

PMID:38796532

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11128000/

Abstract

BACKGROUND

Small extracellular vesicle (sEV) analysis can potentially improve cancer detection and diagnostics. However, this potential has been constrained by insufficient sensitivity, dynamic range, and the need for complex labeling.

METHODS

In this study, we demonstrate the combination of PANORAMA and fluorescence imaging for single sEV analysis. The co-acquisition of PANORAMA and fluorescence images enables label-free visualization, enumeration, size determination, and enables detection of cargo microRNAs (miRs).

RESULTS

An increased sEV count is observed in human plasma samples from patients with cancer, regardless of cancer type. The cargo miR-21 provides molecular specificity within the same sEV population at the single unit level, which pinpoints the sEVs subset of cancer origin. Using cancer cells-implanted animals, cancer-specific sEVs from 20 µl of plasma can be detected before tumors were palpable. The level plateaus between 5-15 absolute sEV count (ASC) per µl with tumors ≥8 mm. In healthy human individuals (N = 106), the levels are on average 1.5 ASC/µl (+/- 0.95) without miR-21 expression. However, for stage I-III cancer patients (N = 205), nearly all (204 out of 205) have levels exceeding 3.5 ASC/µl with an average of 12.2 ASC/µl (±9.6), and a variable proportion of miR-21 labeling among different tumor types with 100% cancer specificity. Using a threshold of 3.5 ASC/µl to test a separate sample set in a blinded fashion yields accurate classification of healthy individuals from cancer patients.

CONCLUSIONS

Our techniques and findings can impact the understanding of cancer biology and the development of new cancer detection and diagnostic technologies.

摘要

背景

小细胞外囊泡（sEV）分析可能会改善癌症检测和诊断。然而，这一潜力受到灵敏度不足、动态范围有限以及需要复杂标记的限制。

方法

在本研究中，我们展示了全景成像（PANORAMA）和荧光成像相结合用于单个sEV分析。同时采集PANORAMA和荧光图像能够实现无标记可视化、计数、尺寸测定，并能检测囊泡携带的微小RNA（miR）。

结果

在癌症患者的人血浆样本中观察到sEV数量增加，无论癌症类型如何。囊泡携带的miR-21在单个单位水平上为同一sEV群体提供分子特异性，从而确定癌症来源的sEV亚群。使用植入癌细胞的动物模型，在肿瘤可触及之前就能检测到20微升血浆中癌症特异性的sEV。当肿瘤≥8毫米时，每微升的绝对sEV计数（ASC）在5 - 15之间趋于平稳。在健康个体（N = 106）中，平均水平为1.5 ASC/微升（±0.95），且无miR-21表达。然而，对于I - III期癌症患者（N = 205），几乎所有患者（205例中的204例）的水平超过3.5 ASC/微升，平均为12.2 ASC/微升（±9.6），并且不同肿瘤类型中miR-21标记的比例各不相同，癌症特异性为100%。以3.5 ASC/微升为阈值，对一个单独的样本集进行盲法检测，能够准确区分健康个体和癌症患者。