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CTC RNA, ccfDNA and gDNA extraction and analysis from a single blood sample, , PreAnalytiX
High demand for liquid biopsy tests calls for combined approaches that allow multimodal analysis of circulating tumor cells (CTCs), circulating cell-free DNA (ccfDNA) and leukocyte-derived genomic DNA (gDNA) from a single blood sample. In particular, analysis of CTC RNA is sensitive to stabilization and storage of blood samples. Join my webinar and learn how the non-crosslinking stabilization reagent of PAXgene Blood ccfDNA Tubes* overcomes these challenges.
* The workflow presented in this webinar is for Research Use Only. Not for use in diagnostic procedures.
Gain new insights by discovering the power of CTCs, , QIAGEN.
The release of CTCs from tumors into blood is driven by the epithelial to mesenchymal transition (EMT) phenomenon, and their survival and metastatic capacity are correlated with a phenotype change into so-called tumor stem cells. Furthermore, molecular characterization of CTCs enables insights into drug resistance mechanisms, changes in mutational profile, and adaptive survival strategies. QIAGEN's AdnaTest can give you added insights that may help you make your next big discovery.
Single-cell CTC analysis, , University Hospital of Essen
In this Science Talk, introduced by Siegfried Hauch and joined by Norbert Hochstein, two alternative workflows for single-cell CTC analysis from blood samples of patients with metastatic breast cancer (MBC) and primary ovarian cancer (POC) are presented, starting with single-cell CTC isolation followed by either a whole transcriptome amplification (WTA) and targeted next-generation sequencing (NGS) workflow or a single-cell transcriptome analysis workflow applying 3' RNA sequencing for high-throughput gene expression analysis.
Be the one of the first to use our new AllPrep DNA/mRNA Nano Kit: Apply for a free field test kit by sending an email to the QIAGEN Cancer Research Team.
AllPrep DNA/mRNA Nano Kit for isolation of DNA and mRNA from very low numbers of cells (rare cells/CTCs):
Send us your Questions we will give our best answer
Working with cell-free DNA, extracellular vesicles or CTCs? Have you ever asked yourself:
ELIMA stands for: Evaluation of Multiple Liquid Biopsy Analytes In Metastatic Breast Cancer Patients All from One Blood Sample
Abstract: Cancer patients are usually treated according to the so-called predictive markers on their primary tumors. However, these markers can change during the molecular evolution of distant metastases, finally leading to death. Primary tumor tissue is only available at the time of first diagnosis and metastatic tissue is often difficult to obtain. Even if accessible, a single tumor biopsy is mostly a single snapshot in time and does not always mirror the characteristics of the disease, mostly due to the known heterogeneity of individual lesions. In this regard, the use of biological fluids such as blood as a non-invasive source of cellular and nucleic acid biomarkers would be an ideal "surrogate tissue" to identify and monitor prognostic and predictive factors that will help in selecting the optimal therapeutic strategy for each individual patient. We approached this challenge and compared and analyzed, all from one blood sample, RNA profiles enclosed in circulating tumor cells or extracellular vesicles and performed mutational analysis of cell-free DNA (cfDNA) in plasma samples (next-generation sequencing) in the follow up of the disease to get insights into their feasibility for therapy stratification and to predict therapeutic options.
Abstract:Cancer patients are usually treated according to the so-called predictive markers on Blood analytes as liquid biopsies, are discussed to be surrogate markers for therapy stratification. Furthermore, the analysis of RNA enclosed in circulating tumor cells (CTCs) or extracellular vesicles (EVs) may be sensitive enough to detect disease progression earlier than contemporary visual staging methods. Repeated analysis is enabled by the minimal invasive nature of blood draw. Here we compared RNA profiles of CTCs and EVs in metastatic breast cancer (MBC) patients to get insights about their feasibility for therapy stratification.
Expression profiling in CTCs as well as in EVs is enabled by the described workflows and may be an alternative for therapy monitoring. Data indicated great differences in RNA profiles of EVs and CTCs.
Abstract:Blood analytes are discussed to be surrogate markers for therapy stratification, as serial sampling enabled by the minimal invasive nature of blood draw facilitates monitoring of clonal evolution. Cell-free DNA (cfDNA) is described to mirror intra-tumoral heterogeneity and gives insights about clonal evolution for improved therapeutic decisions. We sequenced cfDNA of a hormone receptorpositive, HER2-negative metastatic breast cancer (MBC) cohort with a high coverage to examine the prevalence and relevance of any detected variant.
Unique molecular identifiers enable the identification of true positive mutations in cfDNA. The identification of new variants with high prevalence, prognostic value, and dynamics under treatment by deep sequencing of cfDNA might empower sensitive monitoring and personalized therapeutic decisions in the future.
Abstract:Liquid biopsy analytes such as cell-free DNA (cfDNA) and circulating tumor cells (CTCs) exhibit great potential for personalized treatment. Since cfDNA and CTCs are considered to give additive information and blood specimens are limited, isolation of cfDNA and CTC in an “all from one tube” format is desired. We investigated whether cfDNA variant sequencing from CTC-depleted blood impacts the results compared to cfDNA variant sequencing from matched whole blood (WB).
We present a feasible workflow for CTC and ctDNA evaluation for expression and mutation analysis from the same blood sample. Matched cfDNA mutational and CTC transcriptional analyses might empower a comprehensive liquid biopsy analysis to enhance the identification of actionable targets for individual therapy strategies of MBC patients in the future.
Abstract: In this live discussion with QIAGEN research experts, we will summarize the findings of the ELIMA study that have been described in detail in the podcast and science talks over the course of this Cancer Insights Session. We will also look at each part of the workflow for analyzing extracellular vehicles, circulating tumor cells and cfDNA in parallel, and the tools we can use for further analysis of the data. This will include targeted DNA sequencing using UMIs, bioinformatics solutions, targeted methylated-DNA sequencing, miRNA sequencing and simultaneous DNA and RNA profiling by NGS. We will put a spotlight on collection and stabilization of blood samples to enable multianalyte approaches in liquid biopsy and will discuss the need for standardization of methods for validation of biomarkers from basic research to clinical research – standardization is key. Throughout the discussion you will be invited to ask questions using the chat function, or you can ask questions in advance by email.
† QIAGEN products shown here are intended for molecular biology applications. These products are not intended for the diagnosis, prevention or treatment of a disease.
The reference to patients in this podcast refers to the source of clinical samples used for scientific insight. No diagnostic or therapeutic action was taken based on the research findings in this study.
For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at www.QIAGEN.com or can be requested from QIAGEN Technical Services or your local distributor.
‡ This offer is valid until May 4, 2020 and only when order is placed online through QIAGEN web shop, your organization’s purchasing system, your sales rep, or by email using the ELIMA promo code. Offer only applies to selected products listed above and is available only to registered members of the QIAGEN Cancer Research Community. Offer not valid in all countries. Offer is subject to availability of products in respective countries. Offer may not be combined with any other promotions. Void where prohibited. It is the customer's responsibility to ensure that acceptance of this offer will not violate any internal policies of their organization. Offer limited to one order per person. Products in offer are for use in molecular biology research applications only. Not for use in diagnostic procedures.