A Photoacoustic Contrast Agent for miR-21 via NIR Fluorescent Hybridization Chain Reaction,Bioconjugate Chemistry

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A Photoacoustic Contrast Agent for miR-21 via NIR Fluorescent Hybridization Chain Reaction
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2021-08-18 , DOI: 10.1021/acs.bioconjchem.1c00375
Raina M Borum ₁ , Colman Moore ₁ , Soo Khim Chan ₁ , Nicole F Steinmetz _{1,

2,

3,

4,

5,

6} , Jesse V Jokerst _{1,

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Affiliation

Nucleic acids are well-established biomarkers of cancer with immense value in diagnostics and basic research. However, strategies to monitor these species in tissue can be challenging due to the need for amplification of imaging signal from low analyte concentrations with high specificity. Photoacoustic (PA) imaging is gaining traction for molecular imaging of proteins, small biomolecules, and nucleic acids by coupling pulsed near-infrared (NIR) excitation with broadband acoustic detection. This work introduces a PA nucleic acid contrast agent that harnesses NIR fluorophore and quencher-tagged hybridization chain reaction (HCR) for signal amplification. This HCR probe was designed to enable contact quenching between NIR dye-quencher pairs by coercing their direct alignment when miR-21, a microRNA cancer biomarker, is detected. The probe demonstrated a ratiometric PA limit of detection of 148 pM miR-21, sequence specificity against one- and two-base mutations, and selectivity over other microRNAs. It was further tested in live human ovarian cancer (SKOV3) and noncancerous (HEK 293T) cells to exemplify in situ PA activation based on differences in endogenous miR-21 regulation (p = 0.0002). The probe was lastly tested in tissue mimicking phantoms to exemplify sustained contrast in centimeter-range depths and 85.3% photostability after 15 min of laser irradiation. The probe’s miR-21-specific activation and its ability to maintain contrast in biologically relevant absorbing and scattering media support its consideration for live-cell PA microscopy and potential cancer diagnostics. Results from this probe also underscore the combined detection power between ratiometric PA signaling and strand amplification for more sensitive DNA-based PA sensors.

中文翻译：

通过 NIR 荧光杂交链反应的 miR-21 光声对比剂

核酸是公认的癌症生物标志物，在诊断和基础研究中具有巨大价值。然而，由于需要以高特异性放大来自低分析物浓度的成像信号，因此监测组织中这些物种的策略可能具有挑战性。通过将脉冲近红外 (NIR) 激发与宽带声学检测耦合，光声 (PA) 成像在蛋白质、小生物分子和核酸的分子成像方面越来越受欢迎。这项工作介绍了一种 PA 核酸造影剂，该造影剂利用 NIR 荧光团和猝灭剂标记的杂交链式反应 (HCR) 进行信号放大。这种 HCR 探针旨在通过在检测到 miR-21（一种 microRNA 癌症生物标志物）时强制它们直接对齐来实现 NIR 染料-猝灭剂对之间的接触猝灭。该探针展示了 148 pM miR-21 的比例 PA 检测限、针对一个和两个碱基突变的序列特异性以及对其他 microRNA 的选择性。它在活的人卵巢癌 (SKOV3) 和非癌性 (HEK 293T) 细胞中进行了进一步测试，以举例说明基于内源性 miR-21 调节差异的原位PA 激活 ( p = 0.0002)。该探针最后在组织模拟幻影中进行了测试，以举例说明在激光照射 15 分钟后，厘米范围深度的持续对比度和 85.3% 的光稳定性。该探针的 miR-21 特异性激活及其在生物学相关吸收和散射介质中保持对比度的能力支持其用于活细胞 PA 显微镜和潜在癌症诊断的考虑。该探针的结果还强调了比例 PA 信号和链放大之间的组合检测能力，用于更灵敏的基于 DNA 的 PA 传感器。

更新日期：2021-08-18

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