This year the SelectBIO Annual Circulating Biomarkers World Congress 2019 took place on Coronado Island in San Diego, California. Researchers from all over the world who are interested in the most up-to-date technologies gathered to share and advance their expertise on the various classes of circulating biomarkers and their emerging applications.
Visitors were able to listen to over 60 different keynote presentations focusing on multiple topics regarding exosomes and microvesicles, circulating nucleic acids, such as circulating cell-free DNA (cfDNA) and RNAs, as well as circulating tumor cells (CTCs) and cancer stem cells (CSCs). Speakers from academia, as well as industry, addressed technologies for single vesicle isolation, characterization and analysis and also focused on the role of extracellular vesicles (EVs) in cancer and other diseases. The importance of EVs as potential biomarkers in therapeutic and diagnostic technologies was discussed. High-level panel discussions about “Circulating Biomarkers & Liquid Biopsy” with experts from the John Wayne Cancer Institute, Natera, Pfizer and QIAGEN highlighted the in-depth interest of the scientific community at this congress.
Paving the way for liquid biopsy
As one of the biggest sponsors of the congress, QIAGEN presented new workflow solutions for biomarker analysis, including manual and automated sample processing solutions for all major liquid biopsy samples – crossing the bridge from discovery to insight.
Isolation of high-quality RNA from urinary EVs
Isolation of circulating cell-free biomarkers from different types of body fluids poses diverse challenges that need to be addressed. For isolation of exosomal RNA from urine, QIAGEN recently released new solutions that include protocols for isolation of short and long RNAs from EVs in urine, or isolation of cell-free miRNA in higher throughput. During his presentation and throughout the poster sessions, Martin Schlumpberger, Associate Director Scientific Applications, presented optimized workflows for isolation of both intact mRNA (and other long RNAs) as well as miRNA (and other short RNA species) from urine and demonstrated their use for biomarker detection. In the research study Martin presented, intact EVs from urine were bound to an affinity membrane in spin-column format and lysed in situ for RNA isolation prior to separation of long- and short-RNA fractions. For analyzing clinical research samples, qRT-PCR was used to quantify prostate cancer-specific TMPRSS2:ERG (T2:E) fusion transcripts and compared to expression of KLK3 (PSA) in 20 ml urine from 16 individuals scheduled for radical prostatectomy.
The novel workflow to isolate exosomal RNA from urinary EVs is shown to avoid co-purification of inhibitors from the samples and recover RNA with high reproducibility. Applying the extraction to a clinical research study, T2:E fusion transcripts from prostate cancer can be detected consistently in urine from 10 out of 16 samples, which is the expected frequency for this population.
Single blood draw to insight – a liquid biopsy workflow to analyze several biomarkers in parallel
Isolation of urinary exosomes for RNA biomarker discovery was in focus during the three days conference, but isolation and analysis of CTCs were also addressed. To gain comprehensive insights about the complex molecular mechanisms of different individuals with metastatic breast cancer (MBC), Martin demonstrated on his poster how to isolate and analyze CTCs, EVs and cfDNA from the same blood sample in one streamlined workflow. He minimized the volume of blood needed, and minimized sample-to-sample bias while getting complete information about transcriptomic and genomic content.
If you’d like to know more about new methods to detect and characterize CTCs, visit us here.
For purification of exosomal RNA, including miRNA from urine, download the protocol here.
We had three intense and interesting days at the Circulating Biomarker World Congress 2019 in San Diego, California and we’re looking forward to next year’s meeting.