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Identification of prognostic claudins signature in hepatocellular carcinoma from a hepatocyte differentiation model

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Abstract

Background

Loss of terminal differentiation markers and gain of stem cell-like properties are a major hallmark of cancer malignant progression. Identification of novel biomarkers representing tumor developmental progeny and predictive of patients’ prognosis would greatly benefit clinical cancer management.

Methods

Human embryonic stem cells were induced to differentiate into hepatocytes along hepatic lineages. Transcriptomic data from different liver developmental stages were analyzed combining with the RNA-seq data from The Cancer Genome Atlas (TCGA) project. Kaplan–Meier survival analysis and Cox regression analyses were used to analyze the clinical significance in HCC patients.

Results

A shifted expression pattern of claudin (CLDN) family genes were identified to be closely associated with liver development and tumor progression. Claudins with hepatic features were found to be significantly down-regulated and predicted better prognosis in HCC patients. Conversely, another set of claudins with embryonic stem cell features were found to be significantly up-regulated and predicted worse prognosis in HCC patients. A claudin signature score system was further established by combining the two sets of claudin genes. The newly established claudins signature could robustly predict HCC patients’ prognosis in the training, testing, and independent validation cohorts.

Conclusions

In the present study, we developed a novel embryonic developmental claudins signature to monitor the extent of tumor dedifferentiation in HCC from an in vitro hepatocyte differentiation model. The claudins signature might present a great potential in predicting prognostic significance in HCC as cell surface biomarkers, and provide novel therapeutic targets for precision oncology further in the clinic.

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Abbreviations

HCC:

Hepatocellular carcinoma

RNA-seq:

RNA sequencing

TCGA:

The cancer genome atlas

ICGC:

International Cancer Genome Consortium

hESCs:

Human embryonic stem cells

ROC:

Receiver operating characteristic

AUC:

Area under curve

HCS:

Hepatic claudins signature

ECS:

Embryonic claudins signature

CSS:

Claudins signature scores

PPI:

Protein–protein interaction

HR:

Hazard ratio

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Acknowledgements

We thank Prof. Lu and Prof. Lin’s group from the Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China, for kindly establish the in vitro hepatocyte differentiation model for us.

Funding

This work was supported by National Natural Science Foundation of China (81702400); Guangdong Province Universities and Colleges Pear River Scholar Funded Scheme (2018). The funders had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.

Author information

Author notes

  1. Fan-En Kong and Yun-Qiang Tang contributed equally.

Authors and Affiliations

  1. Affiliated Cancer Hospital and Institute of Guangzhou Medical University, Guangzhou, 510095, China

    Fan-En Kong, Yun-Qiang Tang, Yuan-Feng Gong, Mei-Mei Li, Wei Cheng, Hao-Long Li, Wen-Jie Zhu, Shan-Shan Liu, Li Huang, Xin-Yuan Guan, Ning-Fang Ma & Ming Liu

  2. Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, School of Basic Medical Science, Guangzhou Medical University, Guangzhou, 511436, China

    Fan-En Kong, Mei-Mei Li, Wei Cheng, Hao-Long Li, Wen-Jie Zhu, Shan-Shan Liu, Li Huang, Ning-Fang Ma & Ming Liu

  3. Department of Hepatopancreatobiliary Surgery, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China

    Jia-Qiang Mo

  4. General, Visceral and Cancer Surgery, University Hospital of Cologne, Cologne, Germany

    Yue Zhao

  5. Department of Clinical Oncology, State Key Laboratory for Liver Research, The University of Hong Kong, Pok Fu Lam, Hong Kong

    Xin-Yuan Guan

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  1. Fan-En Kong
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Contributions

ML and N-FM initiated and designed the project; F-EK, Y-QT, Y-FG, J-QM, YZ performed statistical analyses; F-EK, Y-QT, M-ML, WC, W-JZ, H-LL performed bioinformatic analyses; S-SL and LH provided the HCC clinical samples and the relevant clinical information; N-FM, and X-YG provided valuable comments; F-EK and ML wrote the manuscript, and all authors reviewed and approved the manuscript.

Corresponding authors

Correspondence to Ning-Fang Ma or Ming Liu.

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Conflict of interest

The authors declare no conflicts of interest in this work.

Ethical approval

The study was approved by the Research Ethics Committees of Affiliated Cancer Hospital & Institute of Guangzhou Medical University. The experiments regarding the establishment of human embryonic stem cells and induced in vitro differentiation were approved and guided by the ethical committee of CITIC-Xiangya Reproductive & Genetic Hospital. The blastocysts, which were donated for the study after obtaining written informed consents from the patients undergoing assisted reproductive technology treatment, came from various clinical useless embryos, including poor quality embryos, abnormal embryos after pre-implantation genetic diagnosis (PGD), zygotes with the abnormal pronuclear number and partheno-activated embryos [11].

Informed consent

Informed consent was obtained from all patients for being included in the study.

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Electronic supplementary material

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12072_2020_10035_MOESM1_ESM.tif

Supplementary file1 Supplementary Fig. S1 Flowchart of selecting claudins signature genes in predicting HCC prognosis (TIF 6588 kb)

12072_2020_10035_MOESM2_ESM.tif

Supplementary file2 Supplementary Fig. S2 Protein interaction of the claudins signatures genes and relevant enrichment analysis. (a) PPI network of claudins signatures genes and the 100 closest interaction genes were formed by using STRING database in Cytoscape 3.6.1. (b) Gene ontology and enrichment analysis also revealed the signaling pathways and function categories significantly associated with claudins signatures related genes, which included pathways or biological processes closely related to embryonic development (TIF 13711 kb)

12072_2020_10035_MOESM3_ESM.tif

Supplementary file3 Supplementary Fig. S3 Forest map (Logistic analysis) of claudins signatures and common clinical character in multiple HCC clinical cohorts. (a) Forest map (Logistic analysis) of hepatic and embryonic claudins signatures and other clinical characteristics in the training cohort (TCGA-LIHC Cohort I, n=189). (b) Similar analysis was presented in the testing cohort (TCGA-LIHC Cohort II, n=182) (c) and validated in an independent validation cohort (LIRI-JP Cohort, n=232). HR less than 1 was considered as low risk and HR more than 1 was considered as high risk. P value less than 0.05 was considered statistically significant. (TIF 10632 kb)

12072_2020_10035_MOESM4_ESM.tif

Supplementary file4 Supplementary Fig. S4 Kaplan-Meier survival analysis of overall survival and disease-free survival in HCC patients treated with sorafenib from the TCGA-LIHC database. (a) The sub-group of patients treated with sorafenib form the TCGA-LIHC database were extracted, and Kaplan-Meier survival analysis was performed according to the “Embryonic claudins signature” (ECS) score (b) “Hepatic claudins signature” (HCS) score, and (c) “Claudins signature score” (CSS) (TIF 946 kb)

12072_2020_10035_MOESM5_ESM.tif

Supplementary file5 Supplementary Fig. S5 Kaplan-Meier survival analysis of overall survival in HCC patients with or without HCV infection from the TCGA-LIHC database (TIF 4125 kb)

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Supplementary file11 (DOCX 51 kb)

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Kong, FE., Tang, YQ., Gong, YF. et al. Identification of prognostic claudins signature in hepatocellular carcinoma from a hepatocyte differentiation model. Hepatol Int 14, 521–533 (2020). https://doi.org/10.1007/s12072-020-10035-z

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  • DOI: https://doi.org/10.1007/s12072-020-10035-z

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