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Exosome Diagnostics, Qiagen Seek to Move Biofluid Nucleic Acid Sample Prep to Research Community

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This article has been updated from a previous to clarify language about Exosome's technology.

Exosome Diagnostics and Qiagen this week announced a partnership to develop and commercialize co-branded kits to capture and process RNA and DNA from biofluid exosomes and other microvesicles.

New York-based Exosome is betting that the agreement, which will initially focus on microRNA isolation, will enable the company to disseminate its proprietary exosome technology to the research community for biomarker discovery applications.

Simultaneously, Exosome is also using its technology in house to develop clinical in vitro diagnostic tests, and believes that partnering with Qiagen on the sample prep front may help it advance these programs more quickly.

Meantime, for Qiagen, the partnership provides it with another novel molecular tool that it may eventually be able to use for clinical applications on its current real-time PCR systems or forthcoming next-generation sequencing platform, although the companies are developing kits to prepare exosome-derived nucleic acids for analysis on any such commercial instruments.

"The exosome industry has really exploded, and the time was right to … put out a kit for the research markets," Exosome CEO James McCullough told PCR Insider this week.

"This will enable researchers now to start doing biomarker discovery," McCullough added. "It was very clear to us from our own clinical IVD programs that … you need to have a very high-quality prep in order to see biomarkers. By partnering with Qiagen, the leader in clinical sample prep, this will be a very powerful tool in the research community to support biomarker discovery."

As part of an active biological packaging and distribution mechanism for RNA and DNA, exosomes (and other microvesicles) and their nucleic acid contents are being investigated for their implications and utility in a broad range of diseases including cancer, central nervous system disorders such as Alzheimer's and Parkinson's diseases, cardiovascular disease, maternal/fetal medicine, and chronic kidney disease.

The natural stability of the exosome compartment allows collection of clinical samples without special tubes or preservatives, according to Exosome. As a result, researchers can perform analysis and biomarker discovery on high-quality RNA from fresh and frozen plasma, serum, urine, and cerebrospinal fluid samples. This is of particular interest for analysis of RNA-based biomarkers such as ALK or RET, the companies said.

Exosome started out from "the end game" of seeking out specific biomarkers from exosomes "and worked our way backward to really understand how you get the highest quality RNA preparation out of the microvesicle fraction," McCullough said.

As such, the company has spent "several years" optimizing this process, he added.

"It's not trivial to understand what a microvesicle is, what an exosome is, and efficiently extract the RNA," McCullough said. "There is a very prescriptive way that you need to do it in order to maintain the integrity of the RNA, otherwise you end up with a compromised preparation. We think we are now putting out the highest quality RNA preparation, which directly correlates to sensitivity around the specific [gene] expression or mutation that you're looking for, and that's the key."

Qiagen's molecular biology and sample prep tools will further enhance this offering. A spokesperson for Qiagen told PCR Insider in an email that Qiagen will be contributing "proprietary filtration devices, other proprietary buffer solutions, and modified sample preparation" to the product-development efforts.

The end result will be commercial research kits that will enable researchers to prepare microRNA, and eventually other types of RNA, from exosomes found in bodily fluids like serum plasma, cerebrospinal fluid, and urine.

The researchers will then be able to take these preparations and interrogate them using "widely available" PCR, pyrosequencing, and next-generation sequencing technologies, allowing them to take repeated, real-time genetic "snapshots" of disease without the need for tissue biopsy, the companies said.

"The key here is the quality of the RNA preparation that you get," McCullough said. "The microvesicle compartment … is a very active biological system that we're dipping into. We're getting a real-time snapshot of RNA regulation in specific diseases. Once you get that pure preparation, then you can take it to all sorts of downstream activities."

The companies plan to launch the first combined products in the first half of 2014. The first application for microRNA isolation will be designed to run on undisclosed automated instrument platforms from Qiagen, which will use its global sales and distribution network to sell the kits.

The product portfolio is also expected to create the basis for development and commercialization of clinical in vitro diagnostic products for a range of non-invasive personalized healthcare products, the companies said.

"Together we expect to create the gold standard in this emerging field of exosome-based testing, advancing research and improving healthcare," Dirk Loeffert, vice president of global product development for life sciences at Qiagen, said in a statement.

"We also intend to co-develop exosome-based workflows for routine use in personalized healthcare, a revolutionary improvement compared to today's widespread dependence on tissue biopsies, offering the ability to create a new dimension of utility for our molecular testing assay portfolio," Loeffert added.

Exosome also has its own exosome-based clinical in vitro diagnostics in development for several disease indications, the most advanced of which is prostate cancer.

"We're doing multi-center IVD studies in conjunction with the Prostate Cancer Foundation," McCullough said. "We fully expect there to be a product offering in 2014."

Researchers led by James McKiernan, a professor of urology and the director of urologic oncology at New York-Presbyterian Hospital and Columbia University Medical Center, presented early data on Exosome's prostate cancer diagnostic, EXO166, at the American Urological Association Annual Meeting in San Diego in May.

In the study, urine samples were collected from patients scheduled to undergo prostate biopsy or radical prostatectomy. Exosomes containing RNA shed into the urine from the prostate were analyzed using Exosome's EXO70 Urine RNA Isolation Kit in conjunction with real-time PCR.

The biopsy patients were divided into two cohorts: the first cohort was tested using a known prostate cancer biomarker, while a novel four-gene prostate cancer signature was tested in the second cohort. Both groups were stratified based on whether their biopsies were positive or negative.

Besides predicting positive biopsy outcome, exosome testing was able to distinguish histologically less-aggressive cancers with lower Gleason scores from those with higher, more aggressive Gleason scores, suggesting that urinary exosome-derived RNA can be used to non-invasively evaluate gene expression in the prostate and accurately predict the likelihood of a positive or negative needle biopsy in addition to distinguishing more aggressive cancers.

"The ability to achieve a stable, high-quality RNA preparation directly from a simple urine sample has significant implications for interrogating the prostate, the bladder, and the kidney on a molecular level, all non-invasively," McKiernan said in a statement this week. He noted that in more than 1,500 patient samples measured in clinical studies, his team has shown that the Exosome technology "enables reproducible, sensitive measure of cancer-specific molecular biomarkers directly from a simple, untreated urine sample."

The partnership with Qiagen may help Exosome advance this and other IVD programs by providing it with proven chemistries and automation.

"When you're talking about converting it to clinical in vitro diagnostics — which need to be manufactured under [good manufacturing practices], and need to have sensitivity and reproducibility for a specific biomarker that you can, say, take to [the US Food and Drug Administration] for approval — that's another step," McCullough said. "That's down the road. But right now we're focused with Qiagen on providing the research community with the highest quality RNA prep that you can get from the microvesicle fraction."