NEW YORK (GenomeWeb) – Johns Hopkins spinout Circulomics this month received a two-year, $1.5 million Phase II Small Business Innovation Research grant from the National Institutes of Health to finalize development of a portfolio of microRNA-profiling technologies that the company hopes will address a need for mid-range multiplex analyses of the small, non-coding RNAs.
Circulomics's first product — an assay kit based on a semi-custom panel of fixed and user-selected miRNAs — is expected to hit the market this year, with additional products rolling out next year and beyond. And while these will initially be for research use only, Circulomics CEO Kelvin Liu told GenomeWeb that the company ultimately hopes to be able to apply its technologies to the clinical diagnostics space.
Circulomics's core technology is dubbed Ligo-miR, which involves a proprietary two-step ligation process to generate encoded miRNA signatures that can be analyzed using different types of instrumentation. According to Liu, the technology is geared for research applications where PCR and microarrays aren't ideal.
While PCR is highly sensitive and works well when studying a small number of miRNAs, "if you want to look at larger panels of five, 10, 15, or 30 microRNAs … across many samples, that quickly becomes a lot of work."
On the other end of the spectrum, microarrays are very effective for analyzing hundreds or thousands of miRNAs relatively quickly, he noted. But for researchers who have identified their miRNAs of interest already and are focused on only 10 or 20 of them, microarrays can be needlessly complex and expensive.
"We saw a void in the marketplace for technologies that can do targeted profiling once you've already identified [miRNAs of interest] … and [want to] screen lots of samples quickly and easily," Liu said.
The company's most advanced product candidate is dubbed Ligo-miR EZ and is geared for routine miRNA profiling in cells and tissues without the need for specialized equipment. Using a thermocycler and polyacrylamide gel electrophoresis (PAGE) apparatus, the product enables 26-plex profiling of 96 samples a day at attomole-level sensitivity.
Ligo-miR EZ kits will feature 19 fixed miRNAs, including five control miRNAs and 12 specific for a particular application, and the user will be able to request five additional miRNAs of their choosing.
Liu said that the first commercially available Ligo-miR EZ kit will likely include miRNAs generally associated with cancer, and that follow-on kits will be specific to particular cancers such as breast and lung. Eventually, Circulomics hopes to offer kits for applications outside of oncology.
He added that Ligo-miR EZ, as well as a DNA/RNA extraction technology called Nanobind that was also developed with NIH funding, are expected to be commercialized this year.
Also under development with the latest NIH grant is Ligo-miR CE, which is targeted to researchers performing high-throughput clinical analyses in single cells and biofluids. This technology involves capillary electrophoresis rather than PAGE in order to increase sample throughput to thousands a day, and includes post-amplification to improve sensitivity to less than 100 total copies. It is designed to analyze 96 single cells at 80-plex coverage using a single 96-well plate.
Ligo-miR CE is expected to reach the market in 2016.
Lastly, Circulomics is developing Ligo-miR HD, which uses the company's PicoSep single-molecular analysis instrument. Because PicoSep works with as little as 5 picoliters of sample and less than 100 DNA molecules, it has near-zero sample consumption, making it ideal for use with rare and limited samples, according to the company.
Ligo-miR HD is expected to have an 18-plex capability and run 24 samples a day. Liu declined to provide a specific timeline for the product's commercialization.
Liu said that Circulomics is aiming to initially introduce all three Ligo-miR products solely for research applications, but the company is hoping to eventually enter the diagnostics field once it has begun generating revenues that can be used to support such an expansion.
Even though a move into the clinic is years away, he said that the company is already laying the groundwork by setting up collaborations with academic groups in order to identify miRNA signatures with diagnostic potential.
"On the clinical side, the most important thing is going to be the biomarker content," he said.