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Journal ArticleDOI

Patient-derived cell line models revealed therapeutic targets and molecular mechanisms underlying disease progression of high grade serous ovarian cancer.

TL;DR: Patient-derived cell lines reveal that PRAME is a potential tumor specific therapeutic target in HGSOC and counteracting the down-regulation of p21 caused by loss of wild-type TP53 might be the key to impede disease progression.
About: This article is published in Cancer Letters.The article was published on 2019-09-10 and is currently open access. It has received 14 citations till now. The article focuses on the topics: PRAME & Ovarian cancer.

Summary (1 min read)

1. Introduction

  • Epithelial ovarian cancer (EOC) is a rare disease with a worldwide incidence and mortality of 240000 and 150000, respectively [1].
  • Standard of care includes debulking surgery and platinum-based therapy.
  • Resistance is considered the major course of the patients’ death [2].
  • Unfortunately, by comparing copy number changes, mutations and mRNA profiles with tumors, it was discovered that the cell lines historically used for ovarian cancer research were most unlikely to be representative of HGSOC [5, 6].
  • To address this challenge, the authors established 34 cell lines from 23 HGSOC patients and analyzed the acquired resistance in these tumors and the molecular mechanisms of disease progression.

2. Materials and methods

  • 1. Establishing and maintaining HGSOC cell lines and primary mesothelial cell culture.
  • The methods of establishing and maintaining cell lines were published previously [11].
  • 2. Extraction of DNA and RNA and cDNA synthesis DNA and RNA were extracted with AllPrep DNA/RNA Mini Kit (QIAGEN, Venlo, The Netherlands).
  • The nbinomTest function of the DEseq R package was used after normalization for the identification of differentially expressed transcripts [15].
  • The PRAME antibody was diluted at 1:100 in Dako Antibody-Diluent and incubated at 4°C overnight.

3. Results

  • 1. HGSOC cell lines and patients 34 tumor cell lines from ascites, tumor tissue and pleural fluid from 23 patients with HGSOC were established (Table 1).
  • All cell lines were authenticated to be from the corresponding patients by DNA finger printing analyses (Supplementary Table S1).
  • The only TP53 wild-type cell line 8540 had a heterozygous KRAS mutation.
  • Some had comparable high expression in both tumor cells as well as in the mesothelial cells (e.g. EGFR, ERBB2, MGAT5, MUC16, SPAG9, TPBG, TSPYL1).
  • A few samples showed lower expression in both samples or in one of the matched samples .

4. Discussion

  • The authors successfully established 34 patient-derived HGSOC cell lines and characterized them at the cellular and molecular level.
  • The authors results also showed that PRAME was not expressed in mesothelial cells lining the peritoneal cavity and in fibroblasts, which makes PRAME an optimal target for therapy development.
  • Endocrine therapy was shown to bring benefits to patients with advanced epithelial ovarian cancer [39], probably by blocking the ESR1 promoted tumor cell growth.
  • The authors cell line models showed that recurrent tumor cells did not become more “resistant” to carboplatin than their primary counterparts, which was in accordance with patient response presented by clinical data, suggesting that recurrent tumors resemble their primary ancestors.
  • Taken together, the authors (i) established and molecularly analyzed 34 cell lines from 23 HGSOC patients, (ii) showed that loss of TP53 wild type was the main driving force of tumorigenesis and tumor progression, and (iii) identified PRAME as a potential therapeutic target.

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Citations
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Journal ArticleDOI
TL;DR: This work focuses on the diverse designer self‐assembling peptide hydrogels for instructive cell constructs in tissue‐specific modeling and the precise oncology remodeling for ovarian cancer, which are issued by several research aspects in a 3D context.
Abstract: Designer self-assembling peptides form the entangled nanofiber networks in hydrogels by ionic-complementary self-assembly. This type of hydrogel has realistic biological and physiochemical properties to serve as biomimetic extracellular matrix (ECM) for biomedical applications. The advantages and benefits are distinct from natural hydrogels and other synthetic or semisynthetic hydrogels. Designer peptides provide diverse alternatives of main building blocks to form various functional nanostructures. The entangled nanofiber networks permit essential compositional complexity and heterogeneity of engineering cell microenvironments in comparison with other hydrogels, which may reconstruct the tumor microenvironments (TMEs) in 3D cell cultures and tissue-specific modeling in vitro. Either ovarian cancer progression or recurrence and relapse are involved in the multifaceted TMEs in addition to mesothelial cells, fibroblasts, endothelial cells, pericytes, immune cells, adipocytes, and the ECM. Based on the progress in common hydrogel products, this work focuses on the diverse designer self-assembling peptide hydrogels for instructive cell constructs in tissue-specific modeling and the precise oncology remodeling for ovarian cancer, which are issued by several research aspects in a 3D context. The advantages and significance of designer peptide hydrogels are discussed, and some common approaches and coming challenges are also addressed in current complex tumor diseases.

72 citations

Journal ArticleDOI
TL;DR: The modulation of PRAME might be useful for the treatment of patients with cancer and this review highlights immunotherapeutic strategies that target PRAME in human malignancies.
Abstract: Preferentially expressed antigen in melanoma (PRAME), which belongs to the cancer/testis antigen (CTA) gene family, plays a pivotal role in multiple cellular processes and immunotherapy response in human cancers. PRAME is highly expressed in different types of cancers and is involved in cell proliferation, apoptosis, differentiation and metastasis as well as the outcomes of patients with cancer. In this review article, we discuss the potential roles and physiological functions of PRAME in various types of cancers. Moreover, this review highlights immunotherapeutic strategies that target PRAME in human malignancies. Therefore, the modulation of PRAME might be useful for the treatment of patients with cancer.

57 citations

Journal ArticleDOI
18 Dec 2019-Cells
TL;DR: It is reported that TIMP-1 acts as a double-edged sword in the EOC microenvironment, directly affecting the response to PT treatment on tumor cells and indirectly altering migration and proliferation of endothelial cells.
Abstract: Epithelial Ovarian Cancer (EOC) is the most lethal gynecological cancer in developed countries, and the development of new strategies to overcome chemoresistance is an awaited clinical need. Angiogenesis, the development of new blood vessels from pre-existing vasculature, has been validated as a therapeutic target in this tumor type. The aim of this study is to verify if EOC cells with acquired resistance to platinum (PT) treatment display an altered angiogenic potential. Using a proteomic approach, we identified the tissue inhibitor of metalloproteinases 1 (TIMP-1) as the only secreted factor whose expression was up-regulated in PT-resistant TOV-112D and OVSAHO EOC cells used as study models. We report that TIMP-1 acts as a double-edged sword in the EOC microenvironment, directly affecting the response to PT treatment on tumor cells and indirectly altering migration and proliferation of endothelial cells. Interestingly, we found that high TIMP-1 levels in stage III–IV EOC patients associate with decreased overall survival, especially if they were treated with PT or bevacizumab. Taken together, these results pinpoint TIMP-1 as a key molecule involved in the regulation of EOC PT-resistance and progression disclosing the possibility that it could be used as a new biomarker of PT-resistance and/or therapeutic target.

20 citations


Cites background from "Patient-derived cell line models re..."

  • ...Recently, a European collaborative group established a large number of cell lines from different patients, different locations, or different times of disease progression, and provided the scientific community with characterized models of the most common subgroup of EOC, namely, the high grade serous ovarian cancer (HGSOC) [5,6]....

    [...]

  • ...Yet, these models, although extremely useful to address research questions regarding the progression of PT-sensitive HGSOC, failed to represent a good model of PT-resistant disease....

    [...]

Journal ArticleDOI
11 Dec 2020-Cancers
TL;DR: It is demonstrated for the first time, that HDAC6 over-expression in HGSOC and all ovarian cancers is a favorable prognostic marker and evidence is provided to suggest that inhibition of HDAC 6 catalytic activity with first generation HDac6 inhibitors has limited efficacy as a monotherapy in H GSOC.
Abstract: Histone deacetylase 6 (HDAC6) is a unique histone deacetylating enzyme that resides in the cell cytoplasm and is linked to the modulation of several key cancer related responses, including cell proliferation and migration. The promising anti-cancer response of the first-generation HDAC6 catalytic inhibitors continues to be assessed in clinical trials, although its role in high grade serous ovarian cancer is unclear. This study investigated HDAC6 tumor expression by immunohistochemistry in high-grade serous ovarian cancer (HGSOC) tissue samples and a meta-analysis of HDAC6 gene expression in ovarian cancer from publicly available data. The pharmacological activity of HDAC6 inhibition was assessed in a patient-derived model of HGSOC. HDAC6 was found to be highly expressed in HGSOC tissue samples and in the patient-derived HGSOC cell lines where higher HDAC6 protein and gene expression was associated with a decreased risk of death (hazard ratio (HR) 0.38, (95% confidence interval (CI), 0.16-0.88; p = 0.02); HR = 0.88 (95% CI, 0.78-0.99; p = 0.04)). Similarly, the multivariate analysis of HDAC6 protein expression, adjusting for stage, grade, and cytoreduction/cytoreductive surgery was associated with a decreased risk of death (HR = 0.19 (95% CI, 0.06-0.55); p = 0.002). Knock-down of HDAC6 gene expression with siRNA and protein expression with a HDAC6 targeting protein degrader decreased HGSOC cell proliferation, migration, and viability. Conversely, the selective inhibition of HDAC6 with the catalytic domain inhibitor, Ricolinostat (ACY-1215), inhibited HDAC6 deacetylation of α-tubulin, resulting in a sustained accumulation of acetylated α-tubulin up to 24 h in HGSOC cells, did not produce a robust inhibition of HDAC6 protein function. Inhibition of HGSOC cell proliferation by ACY-1215 was only achieved with significantly higher and non-selective doses of ACY-1215. In summary, we demonstrated, for the first time, that HDAC6 over-expression in HGSOC and all ovarian cancers is a favorable prognostic marker. We provide evidence to suggest that inhibition of HDAC6 catalytic activity with first generation HDAC6 inhibitors has limited efficacy as a monotherapy in HGSOC.

17 citations

Journal ArticleDOI
08 Aug 2020-Cancers
TL;DR: Ten novel spontaneously immortalized patient-derived ovarian cancer cell lines are described, detailing their respective mutational profiles and gene/biomarker expression patterns, as well as their in vitro and in vivo growth characteristics.
Abstract: Cancer cell lines are amongst the most important pre-clinical models. In the context of epithelial ovarian cancer, a highly heterogeneous disease with diverse subtypes, it is paramount to study a wide panel of models in order to draw a representative picture of the disease. As this lethal gynaecological malignancy has seen little improvement in overall survival in the last decade, it is all the more pressing to support future research with robust and diverse study models. Here, we describe ten novel spontaneously immortalized patient-derived ovarian cancer cell lines, detailing their respective mutational profiles and gene/biomarker expression patterns, as well as their in vitro and in vivo growth characteristics. Eight of the cell lines were classified as high-grade serous, while two were determined to be of the rarer mucinous and clear cell subtypes, respectively. Each of the ten cell lines presents a panel of characteristics reflective of diverse clinically relevant phenomena, including chemotherapeutic resistance, metastatic potential, and subtype-associated mutations and gene/protein expression profiles. Importantly, four cell lines formed subcutaneous tumors in mice, a key characteristic for pre-clinical drug testing. Our work thus contributes significantly to the available models for the study of ovarian cancer, supplying additional tools to better understand this complex disease.

10 citations


Cites background from "Patient-derived cell line models re..."

  • ..., histology and protein biomarker analysis is not available for all the patients from which the cell lines were derived [54,90,96]....

    [...]

References
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Abstract: High-throughput sequencing assays such as RNA-Seq, ChIP-Seq or barcode counting provide quantitative readouts in the form of count data. To infer differential signal in such data correctly and with good statistical power, estimation of data variability throughout the dynamic range and a suitable error model are required. We propose a method based on the negative binomial distribution, with variance and mean linked by local regression and present an implementation, DESeq, as an R/Bioconductor package.

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Journal ArticleDOI
Debra A. Bell1, Andrew Berchuck2, Michael J. Birrer3, Jeremy Chien1  +282 moreInstitutions (35)
30 Jun 2011-Nature
TL;DR: It is reported that high-grade serous ovarian cancer is characterized by TP53 mutations in almost all tumours (96%); low prevalence but statistically recurrent somatic mutations in nine further genes including NF1, BRCA1,BRCA2, RB1 and CDK12; 113 significant focal DNA copy number aberrations; and promoter methylation events involving 168 genes.
Abstract: A catalogue of molecular aberrations that cause ovarian cancer is critical for developing and deploying therapies that will improve patients' lives. The Cancer Genome Atlas project has analysed messenger RNA expression, microRNA expression, promoter methylation and DNA copy number in 489 high-grade serous ovarian adenocarcinomas and the DNA sequences of exons from coding genes in 316 of these tumours. Here we report that high-grade serous ovarian cancer is characterized by TP53 mutations in almost all tumours (96%); low prevalence but statistically recurrent somatic mutations in nine further genes including NF1, BRCA1, BRCA2, RB1 and CDK12; 113 significant focal DNA copy number aberrations; and promoter methylation events involving 168 genes. Analyses delineated four ovarian cancer transcriptional subtypes, three microRNA subtypes, four promoter methylation subtypes and a transcriptional signature associated with survival duration, and shed new light on the impact that tumours with BRCA1/2 (BRCA1 or BRCA2) and CCNE1 aberrations have on survival. Pathway analyses suggested that homologous recombination is defective in about half of the tumours analysed, and that NOTCH and FOXM1 signalling are involved in serous ovarian cancer pathophysiology.

5,878 citations

01 Jun 2011
TL;DR: The Cancer Genome Atlas project has analyzed messenger RNA expression, microRNA expression, promoter methylation and DNA copy number in 489 high-grade serous ovarian adenocarcinomas and the DNA sequences of exons from coding genes in 316 of these tumours as mentioned in this paper.
Abstract: A catalogue of molecular aberrations that cause ovarian cancer is critical for developing and deploying therapies that will improve patients’ lives. The Cancer Genome Atlas project has analysed messenger RNA expression, microRNA expression, promoter methylation and DNA copy number in 489 high-grade serous ovarian adenocarcinomas and the DNA sequences of exons from coding genes in 316 of these tumours. Here we report that high-grade serous ovarian cancer is characterized by TP53 mutations in almost all tumours (96%); low prevalence but statistically recurrent somatic mutations in nine further genes including NF1, BRCA1, BRCA2, RB1 and CDK12; 113 significant focal DNA copy number aberrations; and promoter methylation events involving 168 genes. Analyses delineated four ovarian cancer transcriptional subtypes, three microRNA subtypes, four promoter methylation subtypes and a transcriptional signature associated with survival duration, and shed new light on the impact that tumours with BRCA1/2 (BRCA1 or BRCA2) and CCNE1 aberrations have on survival. Pathway analyses suggested that homologous recombination is defective in about half of the tumours analysed, and that NOTCH and FOXM1 signalling are involved in serous ovarian cancer pathophysiology.

5,609 citations

Journal ArticleDOI
TL;DR: The results indicate that the gap between cell lines and tumours can be bridged by genomically informed choices of cell line models for all tumour types.
Abstract: Cancer cell lines are frequently used as in vitro tumour models. Recent molecular profiles of hundreds of cell lines from The Cancer Cell Line Encyclopedia and thousands of tumour samples from the Cancer Genome Atlas now allow a systematic genomic comparison of cell lines and tumours. Here we analyse a panel of 47 ovarian cancer cell lines and identify those that have the highest genetic similarity to ovarian tumours. Our comparison of copy-number changes, mutations and mRNA expression profiles reveals pronounced differences in molecular profiles between commonly used ovarian cancer cell lines and high-grade serous ovarian cancer tumour samples. We identify several rarely used cell lines that more closely resemble cognate tumour profiles than commonly used cell lines, and we propose these lines as the most suitable models of ovarian cancer. Our results indicate that the gap between cell lines and tumours can be bridged by genomically informed choices of cell line models for all tumour types.

1,150 citations

Journal ArticleDOI
TL;DR: This 'roadmap' for HGSOC was determined after extensive discussions at an Ovarian Cancer Action meeting in January 2015 and aims to reduce incidence and improve outcomes for women with this disease.
Abstract: High-grade serous ovarian cancer (HGSOC) accounts for 70-80% of ovarian cancer deaths, and overall survival has not changed significantly for several decades. In this Opinion article, we outline a set of research priorities that we believe will reduce incidence and improve outcomes for women with this disease. This 'roadmap' for HGSOC was determined after extensive discussions at an Ovarian Cancer Action meeting in January 2015.

801 citations

Frequently Asked Questions (1)
Q1. What are the contributions in "Abstract (185) high grade serous ovarian cancer (hgsoc) is the most frequent type of ovarian cancer. most patients have primary response to platinum-based chemotherapy but frequently relapse, which leads to patient death. a lack of well documented and characterized patient-derived hgsoc" ?

A lack of well documented and characterized patient-derived HGSOC cell lines is so far a major barrier to define tumor specific therapeutic targets and to study the molecular mechanisms underlying disease progression. Particularly, the authors demonstrated that a cancer-testis antigen PRAME and Estrogen Receptor could serve as therapeutic targets. Finally, the authors presented that all HGSOC had no or very low CDKN1A ( p21 ) expression due to loss of wild-type TP53, suggesting that loss of cell cycle control is the determinant for tumorigenesis and progression. In conclusion, patient-derived cell lines reveal that PRAME is a potential tumor specific therapeutic target in HGSOC and counteracting the down-regulation of p21 caused by loss of wild-type TP53 might be the key to impede disease progression.