Multichannel DNA Sensor Array Fingerprints Cell States and Identifies Pharmacological Effectors of Catabolic Processes
Multichannel DNA Sensor Array Fingerprints Cell States and Identifies Pharmacological Effectors of Catabolic Processes
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DOI：
10.1021/acssensors.9b01009
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发表时间：
2019-12-01
期刊：
影响因子：
8.9
通讯作者：
Agasti, Sarit S.
中科院分区：
文献类型：
--
作者：
Das Saha, Nilanjana;Sasmal, Ranjan;Agasti, Sarit S.
发病时的细胞往往与单个或几个分子成分表达水平的细微变化有关，这使得传统使用的生物标记物驱动的临床诊断成为一项具有挑战性的任务。我们在这里展示了一种具有多通道输出的DNA纳米传感器阵列的设计，该阵列基于细胞整体蛋白质组特征的改变来识别细胞的正常或病理状态。将荧光团编码的单链DNA(SsDNA)通过超分子相互作用与表面功能化的金纳米颗粒猝灭剂偶联，形成了这种集成的传感器阵列。在这个设计中，单链DNA序列表现出双重作用，它们既提供与受体金纳米颗粒的不同亲和力，又充当转导元件。分析物分子的独特相互作用模式破坏了非共价超分子络合作用，产生同时的多通道荧光输出，通过基于线性判别分析的机器学习算法实现基于签名的分析物识别。不同类型的细胞，特别是正常细胞和癌细胞，可以利用其荧光指纹有效地区分开来。此外，这种DNA传感器阵列在识别与化学调节分解代谢过程相关的细胞变化方面表现出极高的灵敏度。重要的是，可以调节自噬通量的药理效应器已经通过根据其全球蛋白质签名产生反应而被有效地区分开来。综上所述，这些研究表明，我们的多通道DNA纳米传感器非常适合于快速识别复杂混合物中的细微变化，因此可以很容易地扩展用于临床点诊断、高通量药物筛选或从有限的样本量预测治疗结果。
Cells at disease onset are often associated with subtle changes in the expression level of a single or few molecular components, making traditionally used biomarker-driven clinical diagnosis a challenging task. We demonstrate here the design of a DNA nanosensor array with multichannel output that identifies the normal or pathological state of a cell based on the alteration of its global proteomic signature. Fluorophore-encoded single-stranded DNA (ssDNA) strands were coupled via supramolecular interaction with a surface-functionalized gold nanoparticle quencher to generate this integrated sensor array. In this design, ssDNA sequences exhibit dual roles, where they provide differential affinities with the receptor gold nanoparticle as well as act as transducer elements. The unique interaction mode of the analyte molecules disrupts the noncovalent supramolecular complexation, generating simultaneous multichannel fluorescence output to enable signature-based analyte identification via a linear discriminant analysis-based machine learning algorithm. Different cell types, particularly normal and cancerous cells, were effectively distinguished using their fluorescent fingerprints. Additionally, this DNA sensor array displayed excellent sensitivity to identify cellular alterations associated with chemical modulation of catabolic processes. Importantly, pharmacological effectors, which could modulate autophagic flux, have been effectively distinguished by generating responses from their global protein signatures. Taken together, these studies demonstrate that our multichannel DNA nanosensor is well suited for rapid identification of subtle changes in a complex mixture and thus can be readily expanded for point-of-care clinical diagnosis, high-throughput drug screening, or predicting the therapeutic outcome from a limited sample volume.