Cancer Res Treat.  2019 Jul;51(3):1117-1127. 10.4143/crt.2018.405.

Genetic Profiles Associated with Chemoresistance in Patient-Derived Xenograft Models of Ovarian Cancer

• Li LY
• Kim HJ
• Park SA
• Lee SH
• Kim LK
• Lee JY
• Kim S
• Kim YT
• Kim SW
• Nam EJ

• KMID: 2454303
• DOI: http://doi.org/10.4143/crt.2018.405

Abstract

PURPOSE
Recurrence and chemoresistance (CR) are the leading causes of death in patients with high-grade serous carcinoma (HGSC) of the ovary. The aim of this study was to identify genetic changes associated with CR mechanisms using a patient-derived xenograft (PDX) mouse model and genetic sequencing.
MATERIALS AND METHODS
To generate a CR HGSC PDX tumor, mice bearing subcutaneously implanted HGSC PDX tumors were treated with paclitaxel and carboplatin. We compared gene expression and mutations between chemosensitive (CS) and CR PDX tumors with whole exome and RNA sequencing and selected candidate genes. Correlations between candidate gene expression and clinicopathological variables were explored using the Cancer Genome Atlas (TCGA) database and the Human Protein Atlas (THPA).
RESULTS
Three CR and four CS HGSC PDX tumor models were successfully established. RNA sequencing analysis of the PDX tumors revealed that 146 genes were significantly up-regulated and 54 genes down-regulated in the CR group compared with the CS group. Whole exome sequencing analysis showed 39 mutation sites were identified which only occurred in CR group. Differential expression of SAP25,HLA-DPA1, AKT3, and PIK3R5 genes and mutation of TMEM205 and POLR2A may have important functions in the progression of ovarian cancer chemoresistance. According to TCGA data analysis, patients with high HLA-DPA1 expression were more resistant to initial chemotherapy (p=0.030; odds ratio, 1.845).
CONCLUSION
We successfully established CR ovarian cancer PDX mouse models. PDX-based genetic profiling study could be used to select some candidate genes that could be targeted to overcome chemoresistance of ovarian cancer.

Keyword

Ovarian neoplasms; Patient-derived xenografts; Chemoresistance; RNA sequence analysis; Whole exome sequencing

MeSH Terms

Animals
Carboplatin
Cause of Death
Drug Therapy
Exome
Female
Gene Expression
Genome
Heterografts*
Humans
Mice
Odds Ratio
Ovarian Neoplasms*
Ovary
Paclitaxel
Recurrence
Sequence Analysis, RNA
Statistics as Topic
Carboplatin
Paclitaxel

Figure

• 

  Fig. 1. Different chemoresponses in six ovarian patient-derived xenograft models. Three cases (CS1, CS2, and CS3) showed regression of tumor, three cases (CR1, CR2, and CR3) showed consistent growth of tumor. CS, chemosensitive; CR, chemoresistant; T, paclitaxel; C, carboplatin; ▲, chemotherapy injection.

• 

  Fig. 2. Genetic expression of frequently expressed genes and validation of candidate genes in patient-derived xenograft (PDX). (A) Unsupervised hierarchical clustering analysis using 200 selected genes differentially expressed between chemosensitive (CS) and chemoresistant (CR) groups. FPKM, fragments per kilobase million. (B) Immunohistochemical (IHC) staining of AKT3, PIK3R5, HLA-DPA1, and SAP25 in PDX (×400). Expression levels of AKT3, PIK3R5, HLA-DPA1, and SAP25 were much lower in CS cases than in CR (BC, before chemotherapy; PC, post chemotherapy). (C) IHC staining intensity of AKT3, PIK3R5, HLA-DPA1, and SAP25 in CS and CR group. The protein expression levels of four genes were significantly higher in each CR group in comparison with CS. (D) Quantitative real-time polymerase chain reaction analysis of AKT3, PIK3R5, HLA-DPA1, and SAP25 expression in PDX. The mRNA expression level of four genes were significantly increased in each CR group in comparison with CS group. **p < 0.01, ***p < 0.001.

• 

  Fig. 3. Somatic mutation in five patient-derived xenograft (PDX) models. (A) The proportion of mutational type in five PDX tissues. (B) Number of mutated genes by pathway. The phosphoinositide 3-kinase (PI3K)-AKT and mitogen-activated protein kinase (MAPK) signaling pathways exhibited relatively more mutated genes than other pathways in all PDXs. CR, chemoresistant; CS, chemosensitive.

Reference

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