title: Distinct patterns of B-cell receptor signaling in non-Hodgkin lymphomas identified by single-cell profiling.
abstract: Kinases downstream of B-cell antigen receptor (BCR) represent attractive targets for therapy in non-Hodgkin lymphoma (NHL). As clinical responses vary, improved knowledge regarding activation and regulation of BCR signaling in individual patients is needed. Here, using phosphospecific flow cytometry to obtain malignant B-cell signaling profiles from 95 patients representing 4 types of NHL revealed a striking contrast between chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL) tumors. Lymphoma cells from diffuse large B-cell lymphoma patients had high basal phosphorylation levels of most measured signaling nodes, whereas follicular lymphoma cells represented the opposite pattern with no or very low basal levels. MCL showed large interpatient variability in basal levels, and elevated levels for the phosphorylated forms of AKT, extracellular signal-regulated kinase, p38, STAT1, and STAT5 were associated with poor outcome. CLL tumors had elevated basal levels for the phosphorylated forms of BCR-signaling nodes (Src family tyrosine kinase, spleen tyrosine kinase [SYK], phospholipase Cγ), but had low α-BCR-induced signaling. This contrasted MCL tumors, where α-BCR-induced signaling was variable, but significantly potentiated as compared with the other types. Overexpression of CD79B, combined with a gating strategy whereby signaling output was directly quantified per cell as a function of CD79B levels, confirmed a direct relationship between surface CD79B, immunoglobulin M (IgM), and IgM-induced signaling levels. Furthermore, α-BCR-induced signaling strength was variable across patient samples and correlated with BCR subunit CD79B expression, but was inversely correlated with susceptibility to Bruton tyrosine kinase (BTK) and SYK inhibitors in MCL. These individual differences in BCR levels and signaling might relate to differences in therapy responses to BCR-pathway inhibitors.
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.|Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.|Center for Cancer Biomedicine, University of Oslo, Oslo, Norway.
authors.forename: June H
authors.initials: JH
authors.lastname: Myklebust
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.|Lymphoma Immunotherapy Program, Mount Sinai School of Medicine, New York, NY.
authors.forename: Joshua
authors.initials: J
authors.lastname: Brody
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.
authors.forename: Holbrook E
authors.initials: HE
authors.lastname: Kohrt
authors.affiliation: Cancer Clinic, Department of Oncology, and.
authors.forename: Arne
authors.initials: A
authors.lastname: Kolstad
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.
authors.forename: Debra K
authors.initials: DK
authors.lastname: Czerwinski
authors.affiliation: Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.|Center for Cancer Biomedicine, University of Oslo, Oslo, Norway.|Department of Cellular Therapy, Oslo University Hospital, Oslo, Norway.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Wälchli
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.|Eppley Institute for Cancer Research, University of Nebraska Medical Center, Omaha, NE.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Green
authors.affiliation: Cancer Clinic, Department of Pathology, Oslo University Hospital, Oslo, Norway.
authors.forename: Gunhild
authors.initials: G
authors.lastname: Trøen
authors.affiliation: Center for Cancer Biomedicine, University of Oslo, Oslo, Norway.|Department of Informatics, University of Oslo, Oslo, Norway.
authors.forename: Knut
authors.initials: K
authors.lastname: Liestøl
authors.affiliation: Cancer Clinic, Department of Pathology, Oslo University Hospital, Oslo, Norway.
authors.forename: Klaus
authors.initials: K
authors.lastname: Beiske
authors.affiliation: Service d'Hématologie Clinique, Centre Hospitalier Universitaire de Rennes, Rennes, France.|INSERM U917, Université de Rennes, Rennes, France.
authors.forename: Roch
authors.initials: R
authors.lastname: Houot
authors.affiliation: Laboratory Medicine and Pathology, University of Toronto, Toronto, ON, Canada; and.
authors.forename: Jan
authors.initials: J
authors.lastname: Delabie
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.
authors.forename: Ash A
authors.initials: AA
authors.lastname: Alizadeh
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.|Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, TN.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Irish
authors.affiliation: Oncology Division, Department of Medicine, Stanford University, Stanford, CA.
authors.forename: Ronald
authors.initials: R
authors.lastname: Levy
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D051925:CD79 Antigens / Q000235:genetics / Q000378:metabolism; D003937:Diagnosis, Differential; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008224:Lymphoma, Follicular / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016403:Lymphoma, Large B-Cell, Diffuse / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020522:Lymphoma, Mantle-Cell / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism; D051966:Phospholipase C gamma / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D011947:Receptors, Antigen, B-Cell / Q000235:genetics* / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D059010:Single-Cell Analysis; D000072377:Syk Kinase / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: D051925:CD79 Antigens; C497883:CD79B protein, human; D007075:Immunoglobulin M; D010750:Phosphoproteins; D011947:Receptors, Antigen, B-Cell; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050799:STAT5 Transcription Factor; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human; C000606896:SYK protein, human; D000072377:Syk Kinase; D019061:src-Family Kinases; D051057:Proto-Oncogene Proteins c-akt; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D051966:Phospholipase C gamma