Patient-derived organoids of lung cancer based on organoids-on-a-chip: enhancing clinical and translational applications

Abstract

Lung cancer is one of the most common malignant tumors worldwide, with high morbidity and mortality due to significant individual characteristics and genetic heterogeneity. Personalized treatment is necessary to improve the overall survival rate of the patients. In recent years, the development of patient-derived organoids (PDOs) enables lung cancer diseases to be simulated in the real world, and closely reflects the pathophysiological characteristics of natural tumor occurrence and metastasis, highlighting their great potential in biomedical applications, translational medicine, and personalized treatment. However, the inherent defects of traditional organoids, such as poor stability, the tumor microenvironment with simple components and low throughput, limit their further clinical transformation and applications. In this review, we summarized the developments and applications of lung cancer PDOs and discussed the limitations of traditional PDOs in clinical transformation. Herein, we looked into the future and proposed that organoids-on-a-chip based on microfluidic technology are advantageous for personalized drug screening. In addition, combined with recent advances in lung cancer research, we explored the translational value and future development direction of organoids-on-a-chip in the precision treatment of lung cancer.

Keywords: drug screening; lung cancer; microfluidic chip; organoids-on-a-chip; patient-derived organoids.

FIGURE 1

The construction of biobanks containing complex biological characteristics of several kinds of tumor subtypes by utilizing different methods. (A): a. Bright-field microscopy images of LCOs. b. Representative images of long-term cultured LCOs. c. Representative images of successful and failed 2D and 3D cultures derived from lung cancers with different tissue composition. d. The establishment rate of each cancer models according to lung cancer subtypes. e. The subtypes of established 80 LCOs for the lung cancer biobank (Kim et al., 2019). “Adapted with permission from (Kim et al., 2019). Copyright The Author(s) 2019.” (B): PDOs of NSCLC recapitulate the parental tumor. a. The PDO phenotype was observed with a bright field microscope. b. Morphological changes of PDOs. c. HE-stained and IHC images of PDOs and their parental tumor tissues (Chen et al., 2020). “Adapted with permission from (Chen et al., 2020). Copyright 2020 American Chemical Society.” (C): Establishment of tumor organoids derived from SCLC. a. Tumor tissues were obtained from SCLC patients and cultured under various conditions to form organoids. b. Brightfield images of SPDTO formation in short-term culture under five different culture conditions (Choo et al., 2021). “Adapted with permission from (Choi S. Y. et al., 2021). Copyright 2021 by the authors.” (D): Procedure for generating 2D PDOs. a. Procedure for generating 2D PDOs. b. Comparison of IC50 values to each TKI between 3D and 2D PDOs. c. DNA chromatograms showing EGFR L747P mutation. d. Scheme for model switching (Kim S. Y. et al., 2021). “Adapted with permission from (Kim S. Y. et al., 2021). Copyright 2021 The author.” (E): Characterization of CTOSs from patients’ lung tumors with H&E staining and immunohistochemistry of E-cadherin, α-SMAand CD68 (Endo et al., 2013). “Adapted with permission from (Endo et al., 2013). Copyright 2013 International Association for the Study of Lung Cancer.“Abbreviations: PDOs, patient-derived organoids; LCOs, lung cancer organoids; NSCLC, non-small cell lung cancer; SCLC, small cell lung cancer; SPDTO, SCLC patient-derived tumor organoid; TKI, tyrosine kinase inhibitor; CTOSs, cancer tissue-originated spheroid; EGFR, epidermal growth factor receptor.

FIGURE 2

Comparison of traditional organoid and organoids on-a-chip in the developments and applications of PDOs. Abbreviations: PDOs, patient-derived organoids; TME, tumor microenvironment.

FIGURE 3

Microfluidic Organoids-on-a-Chip platform. (A): Diagram of the in situ freeze–thaw cycle and the subsequent drug sensitivity test on the SMAR-chip (Liu et al., 2021). “Adapted with permission from (Liu et al., 2021). Copyright 2021by the authors.” (B): Schematic illustration of brain metastatic niche of NSCLC cells and microfluidic device for recapitulating the niche. a. The brain metastatic niche involving bTME with bECs and astrocytes. b. Representative photography and drawing of the microfluidic device with seven channels (Kim et al., 2022). “Adapted with permission from (Kim et al., 2022). Copyright 2022 The Authors.” (C): Schematic diagram of the microfluidic device for lung cancer organoids. a. Patient-derived LCOs were cultured and expanded in Matrigel droplets in 24-well plates. b. The YFR passive microflow device allowed for a uniform distribution of the organoids (Jung et al., 2019). “Adapted with permission from (Jung et al., 2019). Copyright The Royal Society of Chemistry 2019.” (D): Characterization of the InSMAR-chip. a. Schematics of the InSMAR-chip and the cross-section view of the chip. b. Photograph of an InSMAR-chip with a droplet array in the microwells. c. Photographs of the droplets in the microwells. d. Schematics of the reagent delivery methods on the InSMAR-chip (Hu et al., 2021). “Adapted with permission from (Hu et al., 2021). Copyright The Author(s) 2021.” (E): a. A schematic diagram of the experimental principle. b. A schematic diagram and image of the lung-on-a-chip (Tan et al., 2022). “Adapted with permission from (Tan et al., 2022). Copyright 2022 by the authors.” (F): Schematic illustration showing the overall experimental strategy for delivery of miR-497-loaded exosome in an in vitro NSCLC model (Jeong et al., 2020). “Adapted with permission from (Jeong et al., 2020). Copyright The Royal Society of Chemistry 2020.” (G): 3D lung spheroid cultures for evaluation of PDT procedures in microfluidic Lab-on-a-Chip system. a. The fabricated microchip. b. The scheme of cell aggregation and spheroid formation in the microwell. c. The scheme of microchamber arrangement. d. The chip holder with the microchip placed and comparison with the 384-well plate (Zuchowska et al., 2017). “Adapted with permission from (Zuchowska et al., 2017). Copyright 2017 Elsevier B.V.” (H): This model reproduces the organ microenvironment-specific cancer growth, tumor dormancy and response to TKI treatment observed in human patients (Hassell et al., 2017). “Adapted with permission from (Hassell et al., 2017). Copyright 2017 The Authors.” (I): A microengineered in vitro 3D culture platform to produce self-assembled and perfusable microvascular beds (Paek et al., 2019). “Adapted with permission from (Paek et al., 2019). Copyright 2019, American Chemical Society.” (J): Schematic illustration of the multi-organ microfluidic chip, which includes an upstream “lung organ” and three downstream “distant organs” (Xu et al., 2016). “Adapted with permission from (Xu et al., 2016). Copyright 2016, American Chemical Society.” (K): 3D-CMOM platform. a. Schematic diagram displays the functional description of each area of the platform. b. A diagram of the multilayer structure that comprises the platform. c. The details of the function of each hole in the platform. d. Image of the microfluidic platform (Zheng et al., 2021). “Adapted with permission from (Zheng et al., 2021). Copyright 2021, American Chemical Society.” (L): Diagram of the expansion of CTCs from early stage lung cancer patients using a microfluidic co-culture model (Zhang et al., 2014). “Adapted with permission from (Zheng et al., 2021). Copyright 2014 Zhang et al.” Abbreviations: NSCLC, non-small cell lung cancer; bTME, brain tumor microenvironment; bECs, brain endothelial cells; BM, brain metastases; LCOs, lung cancer organoids; PDMS, prepared polydimethylsiloxane; EDTA tube: Anticoagulation tube; CTC, circulating tumor cells; 3D-CMOM, three-dimensional-culture multiorgan microfluidic; PMMA, polymethylmethacrylate; YFR, yarn flow resistor; InSMAR-chip, integrated superhydrophobic microwell array chip; PDT, photodynamic therapy; TKI, tyrosine kinase inhibitor.