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Malignant ascites-derived organoid (MADO) cultures for gastric cancer in vitro modelling and drug screening

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Malignant ascites (MA) is a common manifestation in advanced gastric cancer with peritoneal carcinomatosis and usually indicates a poor prognosis. However, lack of in vitro models that can faithfully recapitulate the characteristics of tumour cells in ascites hinders related researches. Tumour organoids have emerged as a robust in vitro model for tumour research and drug screening. Hence, we aimed to generate a 3-D in vitro organoid cultures from malignant ascites of gastric cancer for disease modelling and drug screening.

Methods

Eleven MADOs were generated from the MA tumour cells of gastric cancer patients. We made comparisons between MADOs and original MA tumour cells in histopathology by immunohistochemistry and genomics by whole-exome sequencing. In order to evaluate MADOs as functional in vitro disease models, we tested whether MADOs could be used for drug sensitivity screens.

Results

Eleven MADO cultures from human gastric cancer were established. MADOs demonstrated divergent growth characteristics and morphologies. MADO cultures preserve the histological architecture, genomic landscape of the corresponding MA tumour cells. MADOs exhibited heterogeneous responses to standard-of-care chemotherapeutics.

Conclusions

We generated MADOs modelling characteristics and mutated genes of MA tumour cells. A broad range of intrinsic MADO response to conventional chemotherapeutics suggests MADOs are amenable to drug screening.

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Availability of data and materials

The data and materials supporting the conclusions of the current study are available from the corresponding author on reasonable request.

Abbreviations

CNV:

Copy number variation

GSEA:

Gene set enrichment analysis

HIPEC:

Hyperthermic intraperitoneal chemotherapy

H&E:

Haematoxylin and eosin

ICGC:

International cancer genome consortium

IHC:

Immunohistochemistry

MA:

Malignant ascites

MADO:

Malignant-ascites derived organoid

NCCN:

National comprehensive cancer Network

PBMC:

Peripheral blood mononuclear cell

PCA:

Principal component analysis

PDO:

Patient-derived organoid SNV: single-nucleotide variants

WES:

Whole-exome sequencing

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Acknowledgements

We thank professor Li Gao for providing assistance in immunohistochemistry staining and Huan Xu, Guanglang Zhu as well as Kaijian Wang for providing help in statistical analysis.

Funding

This work was supported by grants from the National Natural Science Foundation of China (31470826), National Major Scientific and Technological Special Project for “Significant New Drugs Development” (2017ZX09304030) and Scientific Research Program of Shanghai Municipal Science and Technology Commission (19411970700).

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Author notes

  1. Jie Li, Huawei Xu and Lixing Zhang are contributed equally to this work.

Authors and Affiliations

  1. Department of Oncology, Changhai Hospital, Second Military Medical University, Shanghai, 200433, China

    Jie Li, Huawei Xu, Lele Song, Dan Feng, Xiaobo Peng, Meihong Wu, Bin Wang & Xianbao Zhan

  2. Research and Early Development, Shanghai 121Biomed Inc, Shanghai, 200235, China

    Lixing Zhang

  3. Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, Institute of Biomedical Sciences, East China Normal University, Shanghai, 200241, China

    Yang Zou

  4. CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China

    Lixing Zhan

  5. Institute of Radiation Medicine and Fudan University Shanghai Cancer Center, Fudan University, Shanghai, 230032, China

    Guoqiang Hua

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  1. Jie Li
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Contributions

JL: design, collection of samples and data. HX: data analysis and interpretation, manuscript writing. LZ: collection of data, revision of the manuscript. LS and LZ: revision of the manuscript and data analysis. XP, DF, MW and BW: collection of samples and data analysis. YZ: genetic analysis. XZ and GH: conception and design, study supervision, data analysis and revision of the manuscript.

Corresponding authors

Correspondence to Guoqiang Hua or Xianbao Zhan.

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Conflicts of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participate

This study was approved by the Ethics Committee of Changhai Hospital of Second Military Medical University (CHEC2016-157), and written informed consent was obtained from all patients or their guardians.

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Informed consent was obtained from all participants for publication.

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Figure S1.

Presentative images of organoids derived from normal gastro and primary gastric cancer. (A) Organoid derived from normal gastric tissue. (B) Organoid derived from gastric cancer (PDF 384 kb)

Figure S2.

Morphological variation of MADOs and CDH gene mutations. (A) H&E staining of 5 MADOs lines. The top row (P6, P7, P10) shows MADOs with a dense structure, while the bottom row (P3, P9) shows MADOs with a loose structure. (B) Mutations of CDH gene family of 6 MADO lines based on WES. All of these CDH family genes were confirmed as a missense mutation (PDF 6154 kb)

Figure S3.

Additional diagram of genomic analysis, Related to Figure 3. (A) Principal component analysis (PCA) of tumour cells from MA (T, cross-shaped), blood (PBMC, triangle), and MADOs (O, circular) based on the principal component analysis. (B) Hierarchical clustering heat-map visualising the genetic distance of 100 gDNA samples from 6 patients. (C) OncoPlot describing entire recurrent mutated cancer-related genes (PDF 439 kb)

Figure S4.

Dose-response curves and half-maximal inhibitory concentrations (IC50) of 9 MADOs to 7 conventional chemotherapy drugs, related to Figure 4. Dose-response curves of MADOs (P1, P2, P3) and gastric cancer organoids (GC1, GC2, GC3, GC4, GC5, GC6) treated with 7 chemical drugs. GC, gastric cancer organoid (PDF 493 kb)

Table S1.

All clinical data were collected from oncologists. The diagnosis was based on imaging, exploratory surgery and biopsy (gastroscope or percutaneous peritoneal biopsy) (PDF 407 kb)

Supplementary material 6 (PDF 293 kb)

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Li, J., Xu, H., Zhang, L. et al. Malignant ascites-derived organoid (MADO) cultures for gastric cancer in vitro modelling and drug screening. J Cancer Res Clin Oncol 145, 2637–2647 (2019). https://doi.org/10.1007/s00432-019-03004-z

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