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김윤진Yoon-Jin Kim

한양대학교

ACS Nano, Feb 06 2024, | https://doi.org/10.1021/acsnano.3c10421 3D Amplified Single-Cell RNA and Protein Imaging Identifies Oncogenic Transcript Subtypes in B-Cell Acute Lymphoblastic Leukemia

Suyeon Shin ¹, Yoon-Jin Kim¹, Hyo Geun Yun¹, Haerim Chung ², Hyunsoo Cho ², Sungyoung Choi ^1,3,4

¹Department of Electronic Engineering, Hanyang University, Seoul 04763, Republic of Korea.

²Division of Hematology, Department of Internal Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea.

³Department of Biomedical Engineering, Hanyang University, Seoul 04763, Republic of Korea.

⁴Department of Healthcare Digital Engineering, Hanyang University, Seoul 04763, Republic of Korea.

S.S. and Y.-J.K. contributed equally to this work.

Corresponding Authors : Hyunsoo Cho, Sungyoung Choi

Abstract

Simultaneous in situ detection of transcript and protein markers at the single-cell level is essential for gaining a better understanding of tumor heterogeneity and for predicting and monitoring treatment responses. However, the limited accessibility to advanced 3D imaging techniques has hindered their rapid implementation. Here, we present a 3D single-cell imaging technique, termed 3D digital rolling circle amplification (4DRCA), capable of the multiplexed and amplified simultaneous digital quantification of single-cell RNAs and proteins using standard fluorescence microscopy and off-the-shelf reagents. We generated spectrally distinguishable DNA amplicons from molecular markers through an integrative protocol combining single-cell RNA and protein assays and directly enumerated the amplicons by leveraging an open-source algorithm for 3D deconvolution with a custom-built automatic gating algorithm. With 4DRCA, we were able to simultaneously quantify surface protein markers and cytokine transcripts in T-lymphocytes. We also show that 4DRCA can distinguish BCR-ABL1 fusion transcript positive B-cell acute lymphoblastic leukemia cells with or without CD19 protein expression. The accessibility and extensibility of 4DRCA render it broadly applicable to other cell-based diagnostic workflows, enabling sensitive and accurate single-cell RNA and protein profiling.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-02-20)
연구진국내 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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