Immunoglobulin (Ig) light chain restriction (LCR) detection is an important molecular diagnostic tool in the differential diagnosis that includes lymphoid hyperplasia, atypical lymphoid hyperplasia, chronic inflammation, and B cell neoplasms. LCR can be detected by flow cytometry (FCM) as an abnormal kappa:lambda ratio on the cell surface. However, FCM requires fresh (unfixed) tissue. Some B cell lymphomas lack surface Ig expression detectable by FCM, some demonstrate nonspecific antibody binding. Often only formalin-fixed paraffin-embedded (FFPE) tissue is available for pathologic evaluation. Current methods of LCR identification in FFPE tissue include immunohistochemistry (IHC) and conventional chromogenic in situ hybridization (CISH), which measure kappa or lambda protein and mRNA, respectively. However, both IHC and conventional CISH lack sufficient sensitivity to detect LCR in many non-Hodgkin lymphoma (NHL), including the most common NHL variants - follicular lymphoma, diffuse large B cell lymphoma, small B lymphocytic lymphoma, and extranodal marginal zone lymphoma of MALT type. RNAscope is a novel RNA in situ hybridization technology which has single-molecule detection sensitivity and is fully compatible with FFPE tissue using either chromogenic or fluorescent detection. We have developed a manual bright-field kappa and lambda duplex RNA ISH assay for LCR, which has demonstrated broad applicability to all NHL subtypes. In this project, we will develop a fully automated duplex RNAscope LCR assay with digital pathology interpretation, which should greatly facilitate clinical adoption of this novel assay.

Public Health Relevance

The diagnosis of lymphoma can be difficult based on morphology alone. Detection of restricted immunoglobulin light chain expression is a molecular ancillary tool to resolve ambiguous lymphoproliferative disorders. The proposed product is a fully automated RNA in situ hybridization assay for light chain mRNAs based on the RNAscope technology, which will provide superior sensitivity to current methods.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Small Business Innovation Research Grants (SBIR) - Phase II (R44)
Project #
2R44CA168019-02A1
Application #
8782413
Study Section
Special Emphasis Panel (ZRG1-OTC-H (13))
Program Officer
Lou, Xing-Jian
Project Start
2012-09-14
Project End
2016-08-31
Budget Start
2014-09-26
Budget End
2015-08-31
Support Year
2
Fiscal Year
2014
Total Cost
$834,236
Indirect Cost
Name
Advanced Cell Diagnostics, Inc.
Department
Type
DUNS #
607435869
City
Hayward
State
CA
Country
United States
Zip Code
94545