scholarly journals Evaluating Tumor Evolution via Genomic Profiling of Individual Tumor Spheroids in a Malignant Ascites from a Patient with Ovarian Cancer Using a Laser-aided Cell Isolation Technique

2018 ◽  
Author(s):  
Sungsik Kim ◽  
Soochi Kim ◽  
Jinhyun Kim ◽  
Boyun Kim ◽  
Se Ik Kim ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cells ◽  
Primary Tumor ◽  
Phylogenetic Trees ◽  
Malignant Ascites ◽  
Tumor Evolution ◽  
Single Nucleotide Variants ◽  
Tumor Spheroids ◽  
Isolation Technique ◽  
Somatic Alteration

AbstractBackgroundEpithelial ovarian cancer (EOC) is a silent but mostly lethal gynecologic malignancy. Most patients present with malignant ascites and peritoneal seeding at diagnosis. In the present study, we used a laser-aided isolation technique to investigate the clonal relationship between the primary tumor and tumor spheroids found in the malignant ascites of an EOC patient. Somatic alteration profiles of ovarian cancer-related genes were determined for eight spatially separated samples from primary ovarian tumor tissues and ten tumor spheroids from the malignant ascites using next-generation sequencing.ResultsWe observed high levels of intra-tumor heterogeneity (ITH) in copy number alterations (CNAs) and single-nucleotide variants (SNVs) in the primary tumor and the tumor spheroids. As a result, we discovered that tumor cells in the primary tissues and the ascites were genetically different lineages. We categorized the CNAs and SNVs into clonal and subclonal alterations according to their distribution among the samples. Also, we identified focal amplifications and deletions in the analyzed samples. For SNVs, a total of 171 somatic mutations were observed, among which 66 were clonal mutations present in both the primary tumor and the ascites, and 61 and 44 of the SNVs were subclonal mutations present in only the primary tumor or the ascites, respectively.ConclusionsBased on the somatic alteration profiles, we constructed phylogenetic trees and inferred the evolutionary history of tumor cells in the patient. The phylogenetic trees constructed using the CNAs and SNVs showed that two branches of the tumor cells diverged early from an ancestral tumor clone during an early metastasis step in the peritoneal cavity. Our data support the monophyletic spread of tumor spheroids in malignant ascites.

Clinical relevance of VEGF-receptor status in primary ovarian cancer: A pilot study for future biomarker analyses.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. 5556-5556
Author(s):  
Pauline Wimberger ◽  
Issam Chebouti ◽  
Jan Dominik Kuhlmann ◽  
Rainer Kimmig ◽  
Eberhart Kuhlisch ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cells ◽  
Primary Tumor ◽  
Prognostic Significance ◽  
Multivariable Analysis ◽  
Progression Free Survival ◽  
Cancer Tissue ◽  
Vegf Receptor ◽  
Antiangiogenic Treatment ◽  
Primary Ovarian Cancer

5556 Background: Antiangiogenic treatment in addition to platinum/taxane based chemotherapy was shown to improve progression-free survival in first- and second-line chemotherapy in ovarian cancer patients. Predictive markers for antiangiogenic treatment are of high interest. Therefore we investigated VEGF-receptor (VEGFR) expression as possible biomarker candidate in primary ovarian cancer tissue and its clinical impact. Methods: A total of 82 patients with primary ovarian cancer were enrolled into this study. Primary tumor tissue was analyzed for the expression of VEGFR1, VEGFR2 and VEGFR3 by immunhistochemistry. Moreover, the presence of circulating tumor cells (CTC) in the blood was detected by immunomagnetic enrichment and multiplex reverse transcriptase-polymerase chain reaction (Adnatest Ovarian Cancer, Adnagen). Disseminated tumor cells (DTC) in the BM were identified by immunocytochemistry using the pancytokeratin antibodyA45B/B3 and subsequent automatic detection based on staining and cytomorphology. Results: Positivity for at least one VEGFR was observed in 43% of cases. The positivity rates for VEGFR1, -2 and -3 were 34%, 17% and 27%, respectively. VEGFR-status of the primary tumor neither correlated with established clinicopathogical parameters (age, FIGO-stage, nodal status, grading, histological subtype) nor with the presence of CTC or DTC. In addition, VEGFR-status does not provide prognostic significance in regard to progression-free and overall survival (OS). Nevertheless, a trend was observed that patients, being positive for VEGFR3 at primary diagnosis, were more likely to experience suboptimal debulking (p = 0.074). CTC-positivity after adjuvant therapy significantly correlated with OS, but multivariable analysis showed only residual tumor as prognostic factor for OS. Conclusions: The VEGFR-family, albeit frequently expressed in our patient cohort, provided neither prognostic nor predictive relevance, but could probably be a predictor for debulking efficiency. The implementation of VEGFR-status into future biomarker studies should carefully be considered.

Immunotherapy with the trifunctional antibody removab leads to significant elimination of tumor cells from malignant ascites in ovarian cancer: Results of a phase I/II study

2004 ◽  
Vol 22 (14_suppl) ◽  
pp. 2504-2504 ◽  
Author(s):  
M. Jaeger ◽  
M. A. Stroehlein ◽  
A. Schoberth ◽  
A. Burges ◽  
M. M. Heiss ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Phase I ◽  
Tumor Cells ◽  
Malignant Ascites

Abstract 2217: Primary tumor loss of heterozygosity proximal to M6P/IGF2R locus is predictive for the presence and persistence of disseminated tumor cells in the bone marrow of ovarian cancer patients

2011 ◽  
Author(s):  
Jan D. Kuhlmann ◽  
Heidi Schwarzenbach ◽  
Friedrich Otterbach ◽  
Martin Heubner ◽  
Pauline Wimberger ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Bone Marrow ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Loss Of Heterozygosity ◽  
Primary Tumor ◽  
Disseminated Tumor Cells

Gene modification of primary tumor cells for active immunotherapy of human breast and ovarian cancer

1995 ◽  
Vol 31 ◽  
pp. S35
Author(s):  
Ramila Philip ◽  
Elisa Brunette ◽  
Bryan Clary ◽  
Lydia Kilinski ◽  
Deepa Murugesh ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cells ◽  
Primary Tumor ◽  
Active Immunotherapy ◽  
Human Breast ◽  
Gene Modification ◽  
Breast And Ovarian Cancer

scholarly journals Neoantigens elicit T cell responses in breast cancer

2021 ◽  
Author(s):  
Takafumi Morisaki ◽  
Makoto Kubo ◽  
Masayo Umebayashi ◽  
Poh Yew ◽  
Sachiko Yoshimura ◽  
...  
Keyword(s):  
Breast Cancer ◽  
T Cell ◽  
Tumor Cells ◽  
Ex Vivo ◽  
T Cell Responses ◽  
Malignant Ascites ◽  
Single Nucleotide Variants ◽  
Synonymous Mutations ◽  
Cell Responses ◽  
Immune Related Genes

Abstract Neoantigens are tumor-specific antigens that arise from non-synonymous mutations in tumor cells. However, their effect on the immune responses in tumor microenvironment are still unclear in breast cancer.We performed whole exome and RNA sequencing of 31 fresh breast cancer tissues and neoantigen prediction on the non-synonymous single nucleotide variants (nsSNVs) among exonic mutations. Neoantigen profiles were determined by predictive HLA binding affinity (IC50<500nM) and mRNA expression with a read count≥1. We evaluated the association between neoantigen load and expression levels of immune-related genes. Moreover, using primary tumor cells established from a breast cancer patient with malignant ascites, we tried to induce cytotoxic T lymphocytes (CTLs) by co-culturing neoantigen peptide-pulsed dendritic cells (DCs) and autologous peripheral lymphocytes. The functions of CTLs were examined by cytotoxicity and IFN-γ ELISpot assay.. Neoantigen load ranged from 6 to 440 (mean, 95) and was positively correlated with the total number of nsSNVs. Although no associations between neoantigen load and mRNA expressions of T cell markers were observed, but the co-culture of neoantigen-pulsed DCs and lymphocytes successfully induced CTLs ex vivo. These results suggest that neoantigen analysis may show utility in developing strategies to elicit T cell responses.

scholarly journals Carrier-Free Multifunctional Nanomedicine For Intraperitoneal Disseminated Ovarian Cancer Therapy

2022 ◽  
Author(s):  
Xiuyu Huang ◽  
Miaojuan Qiu ◽  
Tianqi Wang ◽  
Binbin Li ◽  
Shiqiang Zhang ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cell ◽  
Tumor Cells ◽  
Self Assembly ◽  
Gynecological Cancer ◽  
Advanced Ovarian Cancer ◽  
Malignant Ascites ◽  
Tumor Cell Migration ◽  
Average Size

Abstract Background: Ovarian cancer is the most lethal gynecological cancer which is characterized by extensive peritoneal implantation metastasis and malignant ascites. Despite advances in diagnosis and treatment in recent years, the five-year survival rate is only 25 - 30%. Therefore, developing multifunctional nanomedicine with abilities of promoting apoptosis and inhibiting migration on tumor cells would be a promising strategy to improve the antitumor effect.Methods and results: In this study, we developed a novel ACaT nanomedicine composed of alendronate, calcium ions and cyclin-dependent kinase 7 (CDK7) inhibitor THZ1. With the average size of 164 nm and zeta potential of 12.4 mV, the spherical ACaT nanoparticles were selectively internalized by tumor cells and effectively accumulated in the tumor site. Results of RNA-sequencing and in vitro experiments showed that ACaT promoted tumor cell apoptosis and inhibited tumor cell migration by arresting the cell cycle, increasing ROS and affecting calcium homeostasis. Weekly intraperitoneally administered of ACaT for 8 cycles significantly inhibited the growth of tumor and prolonged the survival of intraperitoneal xenograft mice.Conclusion: In summary, this study presents a new self-assembly nanomedicine with favorable tumor targeting, antitumor activity and good biocompatibility, providing a novel therapeutic strategy for advanced ovarian cancer.

scholarly journals Image-Based Identification and Genomic Analysis of Single Circulating Tumor Cells in High Grade Serous Ovarian Cancer Patients

Cancers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 3748
Author(s):  
Carolin Salmon ◽  
Janina Levermann ◽  
Rui P. L. Neves ◽  
Sven-Thorsten Liffers ◽  
Jan Dominik Kuhlmann ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Genomic Analysis ◽  
Type A ◽  
Tumor Evolution ◽  
High Grade ◽  
C Cells ◽  
Type B ◽  
A Cells

In Ovarian Cancer (OC), the analysis of single circulating tumor cells (sCTCs) might help to investigate genetic tumor evolution during the course of treatment. Since common CTC identification features failed to reliably detect CTCs in OC, we here present a workflow for their detection and genomic analysis. Blood of 13 high-grade serous primary OC patients was analyzed, using negative immunomagnetic enrichment, followed by immunofluorescence staining and imaging for Hoechst, ERCC1, CD45, CD11b and cytokeratin (CK) and sCTC sorting with the DEPArrayTM NxT. The whole genome of single cells was amplified and profiled for copy number variation (CNV). We detected: Type A-cells, epithelial (Hoechstpos, ERCC1pos, CD45neg, CD11bpos, CKpos); Type B-cells, potentially epithelial (Hoechstpos, ERCC1pos, CD45neg, CD11bpos, CKneg) and Type C-cells, potentially mesenchymal (Hoechstpos, ERCC1pos, CD45neg, CD11bneg, CKneg). In total, we identified five (38.5%) patients harboring sCTCs with an altered CN profile, which were mainly Type A-cells (80%). In addition to inter-and intra-patient genomic heterogeneity, high numbers of Type B- and C-cells were identified in every patient with their aberrant character only confirmed in 6.25% and 4.76% of cases. Further identification markers and studies in the course of treatment are under way to expand sCTC analysis for the identification of tumor evolution in OC.

Immunotherapy with the trifunctional antibody removab leads to significant elimination of tumor cells from malignant ascites in ovarian cancer: Results of a phase I/II study

2004 ◽  
Vol 22 (14_suppl) ◽  
pp. 2504-2504 ◽  
Author(s):  
M. Jaeger ◽  
M. A. Stroehlein ◽  
A. Schoberth ◽  
A. Burges ◽  
M. M. Heiss ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Phase I ◽  
Tumor Cells ◽  
Malignant Ascites

scholarly journals Author Correction: Evaluating Tumor Evolution via Genomic Profiling of Individual Tumor Spheroids in a Malignant Ascites

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Sungsik Kim ◽  
Soochi Kim ◽  
Jinhyun Kim ◽  
Boyun Kim ◽  
Se Ik Kim ◽  
...  
Keyword(s):  
Malignant Ascites ◽  
Tumor Evolution ◽  
Genomic Profiling ◽  
Tumor Spheroids

ESTABLISHMENT AND CHARACTERIZATION OF NEW BREAST AND OVARIAN CANCER CELL LINES AS A MODEL FOR STUDYING CELLULAR PLASTICITY IN VITRO

2016 ◽  
Vol 38 (2) ◽  
pp. 94-100 ◽  
Author(s):  
N Bezdieniezhnykh ◽  
A Lykhova ◽  
N Semesiuk ◽  
R Okhrimenko ◽  
Yu Kudryavets
Keyword(s):  
Ovarian Cancer ◽  
Cell Line ◽  
Tumor Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Primary Tumor ◽  
Ovarian Cancer Cell ◽  
Cancer Cell Line ◽  
Breast And Ovarian Cancer

Aim: To compare biological properties of primary tumor cells isolated from malignant effusion of cancer patients with the same cells of permanent lines established during their long-term cultivation in vitro and to assess the impact of phenotypic conversion that was caused by changes in their microenvironment on their behavioral characteristics. Materials and Methods: The study was performed on primary cell cultures from pleural effusion or ascites of breast and ovarian cancer and permanent cell lines derived from them, namely permanent ovarian cancer cell line I, permanent ovarian cancer cell line II and permanent breast cancer cell line I. Biological characteristics were studied using standard cell culture methods and immunocytochemical assays. Results: Three new cell lines were established from breast and ovarian cancer and cell morphology, migration activity, the kinetics of growth, colony forming activity in semisolid agar and sensitivity to anticancer drug were examined. These characteristics were compared with those of the primary tumor cells. It has been shown that among the primary tumor cells from malignant effusion, cells with mesenchymal characteristics were the most prevalent. Cultivation of primary cancer cells in vitro leads to a phenotypic change of their population: it becomes more homogeneous in morphology with predominantly epithelial-like cells. Also, later after a number of cell doublings in vitro, the cell population changes to include cells primarily with immunophenotypic properties characteristic of epithelial cells. These changes include increase in number of E-cadherin-positive cells and a decrease in number of vimentin and α- smooth muscle actin-positive cells. It was found that significant changes in expression of epithelial-mesenchymal transition associated proteins in cells during their cultivation in vitro in new microenvironment are accompanied by a rapid change in their sensitivity to anticancer drugs. Conclusions: The new breast and ovarian cancer cell lines were established and characterized. The induction of phenotypic transdifferentiation in malignant cells from pleural effusion and ascites can be an important approach for suppressing the progression of neoplastic process.

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