RealSeq Wins $300K NHGRI Grant for RNA Fragment Analysis

NEW YORK – RealSeq Biosciences said on Monday that it has received a Phase I Small Business Innovation Research grant from the National Human Genome Research Institute.

The $300,000 award will help the firm develop its RealSeq-RF technology for preparing libraries of small RNA (sRNA) fragments for analysis with next-generation sequencing and to advance its work in small RNA biomarker discovery.

"The development of RealSeq-RF is a key part of our strategy to excel in the RNA fragmentome field," RealSeq CSO Sergei Kazakov said in a statement. "This new approach advances the capability of RealSeq to allow specific detection of all small RNA types. RealSeq-RF provides researchers with the data they need to better understand the role of small RNAs in health and disease."

The RealSeq-RF platform is the basis for the Santa Cruz, California-based firm's RiboMarker RNA fragment-based diagnostic programs, including for Valley fever.

In the grant abstract, RealSeq said that its technology will help analyze the "full complement of the RNA fragmentome, most of which is represented by sRNA and sRNA fragments that are less than 50 nucleotides in size and possess 3'-phosphate or 2',3'-cyclic phosphate along with 5'-hydroxyl or 5'-phosphate termini." 

The firm plans to develop RealSeq-RF using model synthetic sRNAs then validate the approach by analyzing and comparing the sRNA fragmentome from plasma of healthy donors and leukemia patients, according to the grant abstract.

"Because many types of leukemia show no obvious symptoms early in the disease, the development of minimally invasive, early-stage detection of leukemia would be critical for its successful treatment," the firm added.

The SBIR grant follows a $1.5 million seed financing round closed in July.

RealSeq Secures Seed Financing to Advance the RiboMarkerTM Diagnostic Platform

Santa Cruz, CA: RealSeq Biosciences, Inc., a company developing RNA fragmentomics as real-time diagnostics for disease, today announced the close of its oversubscribed seed round. The investment allows RealSeq to deploy the RiboMarkerTM molecular diagnostics platform toward its first indication, Valley Fever, a fungal disease related to the effects of climate change.

The funding round was co-led by Tech Coast Angels Los Angeles and Tech Coast Angels Orange County.  Other investors include Chemical Angel Network, Angel Star Ventures, members of the Berkeley Angel Network, Virtual Angels Network, MEDA Angels, and TiE SoCal Angels.

“RealSeq’s RNA fragmentomics platform has the potential to achieve new diagnostics capabilities allowing earlier detection supporting more informed treatment decisions and better outcomes,” says John Grant, Ph.D., Tech Coast Angels, Los Angeles.  “The RiboMarker Valley Fever test validates the platform for future partnerships and market opportunities in other areas including oncology.”

“RiboMarkers can dynamically report biological states revolutionizing how we detect, monitor and make treatment decisions in real-time,” says Sergio Barberan-Soler, Ph.D., CEO at RealSeq. “This new funding provides critical support to our technology development and launch objectives over the next 24 months.”

The seed round advances the RiboMarker program initially focusing on debilitating and difficult to detect fungal diseases where early treatment is critical to patient outcomes. Specifically, the areas that the company will focus on are:

·      Validating the RiboMarker platform

·      Advancing the Valley Fever LDT

·      Expanding R&D and business operations

 As part of this funding, John Grant, Ph.D., will be joining the RealSeq board of directors. Dr. Grant is an experienced investor and business advisor with extensive corporate management skills including 30+ years at 3M leading global commercialization, strategic planning, and corporate development in several 3M businesses.  He is active in several investment groups and startup accelerators.

RealSeq Biosciences is a commercial stage company and achieved record revenue in 2022 while generating strong customer validation. The Company’s technology is extensively used by researchers in pharmaceutical and biotechnology companies and academic institutions.

About RealSeq Biosciences: RealSeq Biosciences, Inc. is a privately held biotech company with offices and laboratories in Santa Cruz, California.  The company specializes in developing groundbreaking and innovative technologies focusing on RNA fragmentomics, small RNA analysis, NGS research tools and biomarkers. RealSeq’s visionary approach to RNA fragmentomics-based diagnostics is making development and use of dynamic markers of disease a new reality.

https://www.realseqbiosciences.com

Extracellular RNA cross-talk between spermatozoa and oviductal cells

In a paradigm shift for RNA biology, thanks to the discovery and characterization of extracellular RNA (exRNA), it is now known that RNA plays an important role in cell-to-cell communication. Several groups have reported on the different roles that exRNA communication plays in fertilization. In a recent study using low-bias RealSeq-AC libraries to profile ncRNAs from extracellular vesicles (EVs), Reshi and collaborators reported that spermatozoa alter the cargo of extracellular vesicles from oviductal cells. This study further demonstrates the power of accurate exRNA profiling to further study RNA communication.

This study also dissects the RNA cross-talk between oviduct and spermatozoa. The experiments reported use a method that allows the collection of EVs generated upon direct contact and non-contact interaction of spermatozoa with oviductal cells. Using RealSeq-AC libraries to profile ncRNAs from low-yield RNA preps of isolated EVs, the authors further identified distinct cargo for EVs generated by direct vs non-direct spermatozoa contact. Interestingly, mRNA fragments included as EVs cargo, also varied within the different groups with a high proportion of the RNA comprising intron-derived transcripts. This highlights the importance of profiling RNA fragments to identify changes in cellular transcriptome. Additionally, as stated by the authors when talking about RNA fragments with no known function: “Such material, even if it has no functional role in recipient cells, may therefore comprise a rich pool of biomarkers.”

The main conclusion from the study is that the oviduct is not merely a passageway for spermatozoa but a dynamic organ, where RNA cross-talk between spermatozoa and oviductal epithelial cells plays an important role. The RealSeq-AC technology facilitated this novel finding  by allowing the accurate profiling of exRNAs from low input samples.

MicroRNA biomarkers of equine osteoarthritis

Baker et al. 2022

Osteoarthritis is the most common cause of musculoskeletal disorders in aging horses. Unfortunately, current detection methods only identify later stages of the disease when irreversible cartilage degeneration has already occurred. In a recent publication from The University of Edinburgh, researchers use RealSeq-Biofluids technology to identify promising miRNA biomarkers from synovial samples for the early detection of osteoarthritis. Their sequencing data shows how three miRNAs (miR-25, miR-16 and miR-92a) are significantly downregulated in samples from severe osteoarthritis. RealSeq-Biofluids allowed the authors to detect a record 384 miRNAs in total for horse synovial fluid.

One key finding from the paper was that their small sample size -five mild osteoarthritis and twelve severe osteoarthritis samples- severely limited the sensitivity of their study. The authors further performed a power calculation using the sequencing data to determine that up to 36 samples would be needed to demonstrate a power of 95% (significance level = 0.05). This result further highlights the importance of obtaining a sufficient sample size for any project aimed at identifying accurate biomarkers.

Despite the limitations of a small sample size, the authors studied the role that miR-92a (one of the miRNAs differentially expressed when measured by NGS) has on osteoarthritis development. Gain and loss of function experiments were performed to demonstrate the role of miR-92a in equine osteoarthritis. Ultimately, the authors demonstrated that miR-92a influences chondrogenic and hypertrophic markers that are important for osteoarthritis in equine chondrocytes.

At RealSeq Biosciences we are excited that accurate miRNA profiling using RealSeq products is facilitating biomarker development! Contact us to learn more about our products and services.