An Unbiased CRISPR-Cas9 Screening Method for the Identification of Positive and Negative Regulatory Proteins of Cell Adhesion
Yvonne Thus,
Martin de Rooij,
Roderick Beijersbergen,
Marcel Spaargaren
Affiliations
Yvonne Thus
Department of Pathology, Amsterdam UMC location University of Amsterdam, Amsterdam, The NetherlandsLymphoma and Myeloma Center Amsterdam (LYMMCARE), Amsterdam, The Netherlands, Cancer Biology and Immunology – Target & Therapy Discovery, Cancer Center Amsterdam (CCA), Amsterdam, The Netherlands
Martin de Rooij
Department of Pathology, Amsterdam UMC location University of Amsterdam, Amsterdam, The NetherlandsLymphoma and Myeloma Center Amsterdam (LYMMCARE), Amsterdam, The Netherlands, Cancer Biology and Immunology – Target & Therapy Discovery, Cancer Center Amsterdam (CCA), Amsterdam, The Netherlands
Roderick Beijersbergen
Division of Molecular Carcinogenesis and Oncode Institute, The Netherlands Cancer Institute, Amsterdam, The NetherlandsNKI Robotics and Screening Center and Genomics Core Facility, The Netherlands Cancer Institute, Amsterdam, The Netherlands
Marcel Spaargaren
Department of Pathology, Amsterdam UMC location University of Amsterdam, Amsterdam, The NetherlandsLymphoma and Myeloma Center Amsterdam (LYMMCARE), Amsterdam, The Netherlands, Cancer Biology and Immunology – Target & Therapy Discovery, Cancer Center Amsterdam (CCA), Amsterdam, The Netherlands
Mature B-cell lymphomas are highly dependent upon the protective lymphoid organ microenvironment for their growth and survival. Targeting integrin-mediated homing and retention of the malignant B cells in the lymphoid organs, using the Bruton’s tyrosine kinase (BTK) inhibitor ibrutinib, is a highly efficacious FDA-approved therapy for chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), and Waldenström macroglobulinemia (WM). Unfortunately, a significant subset of patients is intrinsically resistant to ibrutinib or will develop resistance upon prolonged treatment. Here, we describe an unbiased functional genomic CRISPR-Cas9 screening method to identify novel proteins involved in B-cell receptor–controlled integrin-mediated adhesion, which provides novel therapeutic targets to overcome ibrutinib resistance. This screening method is highly flexible and can be easily adapted to identify cell adhesion–regulatory proteins and signaling pathways for other stimuli, adhesion molecules, and cell types.Graphical abstract:
