HAYWARD, Calif.—The U.S. National Cancer Institute (NCI) has awarded a two-year, $1.4-million grant under its Small Business Innovation Research Phase II Program to Advanced Cell Diagnostics Inc. (ACD), a company with a focus on in-situ nucleic acid detection for life-sciences research and clinical diagnosis.
The grant will allow ACD, along with its academic partner Cleveland Clinic, to work toward the development and validation of a diagnostic test based on the company’s proprietary RNAscope technology for discriminating various B-cell non-Hodgkin lymphomas (NHLs) from benign lymphoproliferative diseases. The test would be an important advance in diagnosing B-cell lymphomas due to the shortcomings of conventional methods of establishing clonality in the majority of NHLs.
“Reliable in-situ detection of any RNA in routine clinical specimens has been an unmet need for over 40 years despite many efforts and improvements,” says Dr. Xiao-Jun Ma, chief scientific officer of ACD. “Traditional RNA in-situ hybridization (ISH) techniques are limited to the small fraction of highly expressed genes, leaving 95 percent of the expressed genes undetectable. ACD’s RNAscope technology addresses the need of detecting that 95 percent of the transcriptome.”
RNAscope is said to be the first automated multiplex chromogenic and fluorescent in-situ hybridization platform capable of detecting and quantifying RNA biomarkers in situ at single-molecule sensitivity.
A prime example of the potential of the RNAscope technology is in the detection of Ig kappa/lambda light chain mRNAs, which are expressed at extremely low levels in most B-cell lymphomas, falling into the undetectable 95 percent category for conventional techniques. Clinical laboratory detection of Ig light chain restriction (LCR) is a helpful tool in the differential diagnosis that includes lymphoid hyperplasia, atypical lymphoid hyperplasia, chronic inflammation and B-cell neoplasia.
When fresh tissue is available for examination, LCR can be readily detected as an abnormal kappa/lambda surface immunoglobulin ratio using flow cytometry. However these samples are often unavailable in many clinical settings. Existing solutions, including chromogenic in-situ hybridization and immunohistochemistry (IHC), only address a small fraction of B-cell lymphomas, such as multiple myelomas and those lymphomas with plasmacytic differentiation. ACD’s assay reportedly will be applicable to essentially all B-cell lymphoma variants.
“This breakthrough is achieved through a proprietary probe design and signal amplification strategy that allows robust signal generation for true target detection but not for nonspecific background,” says Ma. “This is in contrast to previous efforts focusing mainly on signal enhancement and little on the background problem.”
“Developing and validating an RNAscope-based diagnostic test is similar to that of the more familiar PCR or IHC-based assays,” he notes. “In some ways, it is actually simpler due to the rapid assay development time (new probes can be had within two weeks) and the assurance of probe sensitivity and specificity.”
“The biggest challenge may be that we need to be more vigilant about how samples are fixed and processed since we are detecting RNA, which is more labile than DNA and protein,” Ma says. “We have developed our technology to be compatible with established standards such as CAP/ASCO guidelines for clinical sample preparation. We also strongly recommend the inclusion of positive and negative control probes in the assay to assess RNA adequacy and sample quality. In our experience, most routinely processed clinical specimens, including archival materials that are more than 10 years old, are adequate for RNAscope staining.”
Cleveland Clinic will provide their expertise in pathology and clinical medicine to guide the development and validation of the assay to help to ensure the final product is well-validated and suited for everyday clinical use by pathologists. Cleveland Clinic will also provide access to patient samples and corresponding reference data generated by standard of care testing.
The Small Business Innovation Research grant is a highly competitive program that encourages domestic small businesses to engage in research and development that holds promise for commercialization. ACD had previously received a one-year Phase I grant and completed the project in 2013, which made it eligible to apply for Phase II funding. The Phase II award will be applied to expenses to cover personnel, materials and supplies and facilities related to the proposed research.
“This award is a further validation of the clinical utility of RNAscope technology,” Dr. Yuling Luo, president and CEO of ACD, said in a news release announcing the receipt of the grant. “We are very pleased that NCI has recognized the diagnostic potential of RNAscope technology and are grateful for its continued support.”
