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Pairing Finger Stick Samples With NGS, Circulogene Joins Liquid Biopsy Field's Growing Ranks

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This article has been updated with additional comments from Circulogene to clarify the company's future plans and its relationship to Atherotech, where its technology was intially developed.

NEW YORK (GenomeWeb) – Hot on the heels of several other new liquid biopsy companies, Circulogene — a new firm offering NGS detection of cancer mutations using finger stick-volume blood samples — announced recently that it has begun commercial operations out of its CLIA-certified Birmingham, Alabama lab.

The company hopes to distinguish itself in a crowding field by enabling testing of much smaller sample volumes than other technologies via a proprietary method for the isolation and enrichment of circulating tumor DNA.

Chen-Hsiung Yeh, Circulogene's chief scientific officer, told GenomeWeb that the company is currently processing samples, though not at a very high volume yet.

The firm's initial offering is a pan-cancer sequencing assay that covers approximately 3,000 hotspot mutations in 50 of the most clinically relevant cancer genes. Though the company runs the same panel for any cancer patient, specific subsets of genes may have more relevance for one or another tumor type. Overall, the test is marketed for melanoma, breast, colorectal, lung, GIST, hematological, ovarian, pancreatic, gastric, and thyroid cancers.

Circulogene's liquid biopsy testing methodology was originally developed by several of the company's founders while employed by cardiovascular testing company Atherotech Diagnostics, Yeh told GenomeWeb.

Previously scientific director of hematology and oncology at Quest Diagnostics, Yeh joined Atherotech to spearhead a new direction for the company in the molecular testing space. Initially, he and other collaborators developed a method for increasing the sensitivity of liquid biopsy analysis in small sample volumes using Atherotech's ultracentrifugation technology in house.

When they launched Circulogene as its own company, the group then further developed their ctDNA isolation technology — using fractionation and enzyme manipulation, Yeh said — which is now the proprietary heart of the firm's liquid biopsy testing service.

Circulogene has demonstrated in research data that its proprietary method can collect more than 100 times more cfDNA while using about one-tenth as much blood as used in the QIAamp Circulating Nucleic Acid kit, a tool used in a variety of other liquid biopsy applications.

"Our competitors in this field … they all need a tube of blood, which is 10 to 20 ml or ccs," Yeh said. "The reason for that is that all these companies are using the same methods to extract or isolate cell-free DNA from plasma, and because of that, they all lose a lot of DNA, so they require more starting material to compensate."

In essence, he argued, you can't detect DNA that isn’t there. "You are losing 70 percent of your starting material, no matter how sensitive your downstream technology, whether that is digital PCR or digital NGS, or other methods trying to reduce the noise background," Yeh said.

In addition to making public its launch last month, Circulogene also announced last week that it is using informatics firm Station X's GenePool platform to analyze and interpret its liquid biopsy sequencing data.

Servicing a clinical diagnostics partner is new for Station X, Richard Goold, the company's CEO and co-founder, told GenomeWeb.

"Initially, we were building this for organizations doing things like biomarker discovery, with large amounts of NGS data. This was a few years ago. And at that point those were the guys with the problem, and those were the main users," Goold said.

"But what we've seen is that clinical applications for genomics have come center stage much quicker than most people anticipated … customers have been coming to us asking if we can do other things as well … and that's actually how we came to Circulogene," he said. "They are hungry to use their technology and [to bring] their test to market quickly, and they didn't want to be messing around with building infrastructure to manage data and build reports."

According to Yeh, the reporting system Station X's platform has allowed includes a breakdown of actionable findings as well as a clear delineation of potential therapies linked to those identified mutations.

"It can help doctors know what kind of mutations are present in what genes and in what percentage," he said, "and it also provides physicians with up to date FDA-cleared drugs against the patient's own cancer type and in other indications, and finally an up to date list of active clinical trials."

Unlike the company's unique sample preparation component, Yeh said its sequencing strategy isn't anything exclusive. The company sequences isolated DNA using Thermo Fisher Scientific's Ion Torrent Proton.

According to Yeh, the company's technical validation experiments have shown that its technology allows for detection of mutations down to a limit of detection of 1 percent allele frequency.

Despite the advantages of being able to extract more DNA from a smaller sample, this sensitivity is actually higher than the lowest limit of detection claimed by some other companies. Guardant Health, for example, reports that its test is sensitive down to a LoD of 0.1 percent.

Circulogene hopes to share additional validation data at next year's annual meeting of the American Association for Cancer Research, including results from deep sequencing of 95 clinical specimens showing high sensitivity, Yeh said.

And though Circulogene only offers a blood-based testing service right now, Yeh said the firm is looking into other sample types as well, such as urine and cerebral spinal fluid. He also noted that an unnamed team of researchers in the UK recently sent the company a difficult CSF sample in which they could not isolate circulating tumor DNA using other methods — but Circulogene was able to successfully extract cell-free DNA from the sample.

According to Yeh, the near-term plan for the company is to focus its business locally. Scott Rezek, the company's co-founder, COO, and head of marketing, clarified though that there are also oncologists and researchers nationwide that use Circulogene's laboratory services.

In the longer term, Yeh said, Circulogene is considering shifting from a laboratory-developed test model to potentially develop kits and seek clearance from the US Food and Drug Administration, but Rezek said that the company has no direct plans in that direction yet.