Microdroplet-enhanced chip platform for high-throughput immunotherapy marker screening from extracellular vesicle RNAs and membrane proteins

Chuanhao Tang; Zaizai Dong; Shi Yan; Bing Liu; Zhiying Wang; Long Cheng; Feng Liu; Hong Sun; Yimeng Du; Lu Pan; Yuhao Zhou; Zhiyuan Jin; Libo Zhao; Nan Wu; Lingqian Chang; Xiaojie Xu

doi:10.1016/j.bios.2024.116748

Microdroplet-enhanced chip platform for high-throughput immunotherapy marker screening from extracellular vesicle RNAs and membrane proteins

Biosens Bioelectron. 2025 Jan 1:267:116748. doi: 10.1016/j.bios.2024.116748. Epub 2024 Sep 10.

Authors

Chuanhao Tang¹, Zaizai Dong², Shi Yan³, Bing Liu³, Zhiying Wang⁴, Long Cheng⁵, Feng Liu⁴, Hong Sun⁶, Yimeng Du⁷, Lu Pan⁷, Yuhao Zhou⁴, Zhiyuan Jin⁴, Libo Zhao⁸, Nan Wu⁹, Lingqian Chang¹⁰, Xiaojie Xu¹¹

Affiliations

¹ Department of Genetic Engineering, Beijing Institute of Biotechnology, Beijing, 100071, China; Department of Medical Oncology, Peking University International Hospital, Beijing, 102206, China.
² School of Engineering Medicine, Beihang University, Beijing, 100191, China; Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191, China. Electronic address: dongzaizai@buaa.edu.cn.
³ State Key Laboratory of Molecular Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Thoracic Surgery II, Peking University Cancer Hospital and Institute, Beijing, 100142, China.
⁴ Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191, China.
⁵ School of Biomedical Engineering, Anhui Medical University, Hefei, 230032, China.
⁶ Translational Medicine Center, Beijing Chest Hospital, Capital Medical University, Beijing Key Laboratory in Drug Resistant Tuberculosis Research, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing, 101149, China.
⁷ Department of Genetic Engineering, Beijing Institute of Biotechnology, Beijing, 100071, China.
⁸ Echo Biotech Co., Ltd, Beijing, 102206, China.
⁹ State Key Laboratory of Molecular Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Thoracic Surgery II, Peking University Cancer Hospital and Institute, Beijing, 100142, China. Electronic address: nanwu@bjmu.edu.cn.
¹⁰ Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191, China; School of Biomedical Engineering, Anhui Medical University, Hefei, 230032, China. Electronic address: lingqianchang@buaa.edu.cn.
¹¹ Department of Genetic Engineering, Beijing Institute of Biotechnology, Beijing, 100071, China. Electronic address: miraclexxj@126.com.

PMID: 39276441
DOI: 10.1016/j.bios.2024.116748

Abstract

Extracellular vesicles (EVs) are considered as promising candidates for predicting patients who respond to immunotherapy. Nevertheless, simultaneous detection of multiple EVs markers still presents significant technical challenges. In this work, we developed a high-throughput microdroplet-enhanced chip (MEC) platform, which utilizes thousands of individual microchambers (∼pL) as reactors, accelerating the detection efficiency of the CRISPR/Cas systems and increasing the sensitivity by up to 100-fold (aM level). Ten biomarkers (including 5 RNAs and 5 proteins) from patients' EVs are successfully detected on one chip, and the comprehensive markers show increased accuracy (AUC 0.911) than the individual marker for the efficacy prediction of immunotherapy. This platform provides a high-throughput yet sensitive strategy for screening immunotherapy markers in clinical.

Keywords: Aptamer; CRISPR/Cas; Extracellular vesicle; Immunotherapy; Microfluidic chip.

MeSH terms

Biomarkers / analysis
Biomarkers, Tumor
Biosensing Techniques* / instrumentation
Biosensing Techniques* / methods
CRISPR-Cas Systems
Equipment Design
Extracellular Vesicles* / chemistry
High-Throughput Screening Assays / instrumentation
Humans
Immunotherapy*
Lab-On-A-Chip Devices
Membrane Proteins*
RNA / genetics

Substances

Membrane Proteins
RNA
Biomarkers
Biomarkers, Tumor