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  2. A facile liquid biopsy assay for highly efficient CTCs capture and reagent-less monitoring of immune checkpoint PD-L1 expression on CTCs with non-small cell lung cancer patients

A facile liquid biopsy assay for highly efficient CTCs capture and reagent-less monitoring of immune checkpoint PD-L1 expression on CTCs with non-small cell lung cancer patients

  • Biosens Bioelectron. 2025 May 1:275:117236. doi: 10.1016/j.bios.2025.117236.
Yuping Liu 1 Beibei Zhang 1 Xueyuan Wu 2 Fan Wang 1 Zhiyi Yang 1 Mengyi Li 1 Kaixuan Sheng 1 Yue Yan 1 Liang Zhu 3 Hui Jing 4 Yanmin Wu 4 Lili Hu 5 Yanyan Yu 6 Chenglin Li 7
Affiliations

Affiliations

  • 1 Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, PR China.
  • 2 Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, PR China; Department of Pharmacy, Xuzhou Tongshan District Traditional Chinese Medicine Hospital, Xuzhou, 221116, Jiangsu, PR China.
  • 3 Department of Pharmacy, The Affiliated Huaian No.1 People's Hospital of Nanjing Medical University, Huaian, 223300, Jiangsu, PR China.
  • 4 Department of Respiratory and Critical Care Medicine, The Xuzhou School of Clinical Medicine of Nanjing Medical University, Xuzhou Central Hospital, Xuzhou, 221009, Jiangsu, PR China.
  • 5 Department of Pharmacy, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, 221004, Jiangsu, PR China.
  • 6 Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, PR China. Electronic address: yyyxzmc@163.com.
  • 7 Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, 221004, Jiangsu, PR China. Electronic address: licl@xzhmu.edu.cn.
Abstract

Programmed cell death 1 ligand 1 (PD-L1) immunotherapy holds a pivotal role in lung Cancer treatment. However, current methods for monitoring PD-L1 expression exhibit several limitations, including hysteresis and the invasive nature of tissue sampling. Circulating tumor cells (CTCs), the important biomarkers in liquid biopsy, are minimally invasive and facilitate continuous monitoring. Consequently, integrating CTC counting with PD-L1 expression analysis offers more comprehensive insights for the development of personalized treatment strategies development and efficacy evaluations. In this study, we presented a facile liquid biopsy assay designed for the dynamic monitoring of PD-L1 expression on CTCs captured from non-small cell lung Cancer (NSCLC) patients. This assay was achieved by fabricating two high-performance probes: EpCAM and Vimentin dual-aptamer modified nitrogen-doped carbon quantum dots probe (E/V-apt-N-CQDs) and hairpin PD-L1 aptamer coupled with gold nanoparticles (PD-L1-apt-AuNPs). The E/V-apt-N-CQDs probe effectively captured two types of CTC models (H1299 and A549) exhibiting differential PD-L1 expression. Additionally, reagent-less detection of PD-L1 levels on CTCs was achieved using a portable magnetic electrochemical sensor with excellent specificity and sensitivity, which was capable of measuring PD-L1 concentrations as low as 2 ng/mL. Finally, this assay was applied in 41 NSCLC patients to investigate the correlation between CTC numbers or PD-L1 expression and disease progression and immunotherapy efficacy. The results indicated a significant association between elevated CTC counts or reduced PD-L1 levels and clinical progression. Moreover, this liquid assay successfully monitored dynamic changes in CTCs and PD-L1 expression in NSCLC patients receiving immunotherapy, indicating its potential for clinical application.

Keywords

CTCs capture; Clinical investigations; Lung cancer; Nitrogen-doped carbon quantum dots; PD-L1 expression; Reagent-less electrochemistry.

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