Terrence T. Roh (University of Illinois Urbana-Champaign, GSK plc) | 한빛사논문 > 한빛사

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University of Illinois Urbana-Champaign, GSK plc

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Redox Biol., Sep 2024, 75 103280 S2213-2317(24)00258-1 | https://doi.org/10.1016/j.redox.2024.103280 Predicting DNA damage response in non-small cell lung cancer organoids via simultaneous label-free autofluorescence multiharmonic microscopy

Terrence T. Roh ^a,b, Aneesh Alex ^a,b, Prasanna M. Chandramouleeswaran ^b, Janet E. Sorrells ^a,e, Alexander Ho ^a,e, Rishyashring R. Iyer ^a,d, Darold R. Spillman Jr. ^a,g, Marina Marjanovic ^a,e,g, Jason E. Ekert^b, BanuPriya Sridharan^b, Balabhaskar Prabhakarpandian ^b, Steve R. Hood ^a,c, Stephen A. Boppart ^a,d,e,f,g

^aGSK Center for Optical Molecular Imaging, Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

^bIn Vitro In Vivo Translation, GSK plc, Collegeville, PA, 19426, USA

^cIn Vitro In Vivo Translation, GSK plc, Stevenage, SG1 2NY, UK

^dDepartment of Electrical and Computer Engineering, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

^eDepartment of Bioengineering, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

^fCancer Center at Illinois, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

^gNIH/NIBIB Center for Label-free Imaging and Multi-scale Biophotonics (CLIMB), University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA

Corresponding author: Stephen A. Boppart

Abstract

The DNA damage response (DDR) is a fundamental readout for evaluating efficacy of cancer therapeutics, many of which target DNA associated processes. Current techniques to evaluate DDR rely on immunostaining for phosphorylated histone H2AX (γH2AX), which is an indicator of DNA double-strand breaks. While γH2AX immunostaining can provide a snapshot of DDR in fixed cell and tissue samples, this method is technically cumbersome due to temporal monitoring of DDR requiring timepoint replicates, extensive assay development efforts for 3D cell culture samples such as organoids, and time-consuming protocols for γH2AX immunostaining and its evaluation. The goal of this current study is to reduce overall burden on assay duration and development in non-small cell lung cancer (NSCLC) organoids by leveraging label-free multiphoton imaging. In this study, simultaneous label-free autofluorescence multiharmonic (SLAM) microscopy was used to provide rich intracellular information based on endogenous contrasts. SLAM microscopy enables imaging of live samples eliminating the need to generate sacrificial sample replicates and has improved image acquisition in 3D space over conventional confocal microscopy. Predictive modeling between label-free SLAM microscopy and γH2AX immunostained images confirmed strong correlation between SLAM image features and γH2AX signal. Across multiple DNA targeting chemotherapeutics and multiple patient-derived NSCLC organoid lines, the optical redox ratio and third harmonic generation channels were used to robustly predict DDR. Imaging via SLAM microscopy can be used to more rapidly predict DDR in live 3D NSCLC organoids with minimal sample handling and without labeling.

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형식Research article
게재일2024년 09월 (BRIC 등록일 2024-08-28)
연구진국외 연구진
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