NCI Awards $1.4 Million Grant to Develop Test for B-Cell NHLs

Advanced Cell Diagnostics, Inc. (ACD) and Cleveland Clinic were jointly awarded a 2-year, $1.4 million grant from the National Cancer Institute (NCI) to develop and validate a diagnostic test to identify B-cell non-Hodgkin lymphomas (NHLs) from benign lymphoproliferative diseases. The test will use ACD’s proprietary in situ hybridization assay, called RNAscope technology.

The grant was awarded to ACD and Cleveland Clinic to develop “a fully automated assay and advanced image analysis algorithms for objective interpretation to facilitate clinical adoption,” according a press release from the company.

“This award is a further validation of the clinical utility of RNAscope technology,” Yuling Luo, PhD, President and CEO of ACD, said in a statement. “We are very pleased that NCI has recognized the diagnostic potential of RNAscope technology and are grateful for its continued support.”

Data valuable to the diagnosis of NHLs can be drawn from the restricted expression of kappa or lambda immunoglobulin light chains. Formalin-fixed paraffin-embedded tissue (FFPE) is typically the only sample type available for diagnostic testing, though it is difficult to detect light chain mRNAs in these tissues.

Conventional immunohistochemistry and chromogenic in situ hybridization are not adequate methods to determine clonality in the majority of NHLs, James Cook, MD, PhD, co-investigator at Cleveland Clinic, said in a statement. Researchers believe RNAscope technology can overcome this challenge that has existed for over 20 years.

With its single-molecule sensitivity and single-cell resolution, RNAscope technology can allow for direct visualization of kappa and lambda mRNA in FFPE tissue section under a standard bright-field microscope.

“In phase I, we developed a manual RNAscope-based assay for light chain mRNAs and demonstrated that it allowed determination of light chain restriction in virtually all types of NHL using FFPE with 99% concordance with the current gold standard assay flow cytometry using fresh tissue,” Xiao-Jun Ma, PhD, ACD’s Chief Scientific Officer and principal investigator, said in a statement.

Clonal expansion of B-lymphocytes with immunoglobulin gene rearrangements is a distinctive feature of a B-cell malignancy, and is commonly manifested as expression of kappa or lambda immunoglobulin light chain mRNAs and proteins.

RNAscope technology uses a novel probe design to amplify target-specific signals and ignore background noise from non-specific hybridization. The design is similar to fluorescence resonance energy transfer, according to ACD, and uses two independent double Z probes to “hybridize to the target sequence in tandem in order for signal amplification to occur.” This design selectively amplifies target-specific signals, as independent probes will not hybridize to non-specific targets next to one another.

This grant was awarded under the NCI’s Small Business Innovation Research (SBIR) Phase II program. Phase II SBIR grants are typically given in the amount of $1 million over 2 years, but with justification to complete the project, can exceed that amount over a longer period.

RNAscope technology has been explored in other human cancers, such as breast cancer, head and neck squamous cell carcinoma, phosphaturic mesenchymal tumors, non-small cell lung cancer, and more.