An aberrant NOTCH2-BCR signaling axis in B cells from patients with chronic GVHD

Blood. 2017 Nov 9;130(19):2131-2145. doi: 10.1182/blood-2017-05-782466. Epub 2017 Aug 29.

Abstract

B-cell receptor (BCR)-activated B cells contribute to pathogenesis in chronic graft-versus-host disease (cGVHD), a condition manifested by both B-cell autoreactivity and immune deficiency. We hypothesized that constitutive BCR activation precluded functional B-cell maturation in cGVHD. To address this, we examined BCR-NOTCH2 synergy because NOTCH has been shown to increase BCR responsiveness in normal mouse B cells. We conducted ex vivo activation and signaling assays of 30 primary samples from hematopoietic stem cell transplantation patients with and without cGVHD. Consistent with a molecular link between pathways, we found that BCR-NOTCH activation significantly increased the proximal BCR adapter protein BLNK. BCR-NOTCH activation also enabled persistent NOTCH2 surface expression, suggesting a positive feedback loop. Specific NOTCH2 blockade eliminated NOTCH-BCR activation and significantly altered NOTCH downstream targets and B-cell maturation/effector molecules. Examination of the molecular underpinnings of this "NOTCH2-BCR axis" in cGVHD revealed imbalanced expression of the transcription factors IRF4 and IRF8, each critical to B-cell differentiation and fate. All-trans retinoic acid (ATRA) increased IRF4 expression, restored the IRF4-to-IRF8 ratio, abrogated BCR-NOTCH hyperactivation, and reduced NOTCH2 expression in cGVHD B cells without compromising viability. ATRA-treated cGVHD B cells had elevated TLR9 and PAX5, but not BLIMP1 (a gene-expression pattern associated with mature follicular B cells) and also attained increased cytosine guanine dinucleotide responsiveness. Together, we reveal a mechanistic link between NOTCH2 activation and robust BCR responses to otherwise suboptimal amounts of surrogate antigen. Our findings suggest that peripheral B cells in cGVHD patients can be pharmacologically directed from hyperactivation toward maturity.

Publication types

  • Clinical Trial
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / biosynthesis
  • Adaptor Proteins, Signal Transducing / genetics
  • Adult
  • Aged
  • Allografts
  • B-Lymphocytes / metabolism*
  • B-Lymphocytes / pathology
  • Chronic Disease
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Graft vs Host Disease / genetics
  • Graft vs Host Disease / metabolism*
  • Graft vs Host Disease / pathology
  • Hematologic Neoplasms / genetics
  • Hematologic Neoplasms / metabolism
  • Hematologic Neoplasms / pathology
  • Hematopoietic Stem Cell Transplantation*
  • Humans
  • Interferon Regulatory Factors / biosynthesis
  • Interferon Regulatory Factors / genetics
  • Male
  • Middle Aged
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • Receptor, Notch2 / genetics
  • Receptor, Notch2 / metabolism*
  • Receptors, Antigen, B-Cell / genetics
  • Receptors, Antigen, B-Cell / metabolism*
  • Signal Transduction*
  • Tretinoin / pharmacology

Substances

  • Adaptor Proteins, Signal Transducing
  • B cell linker protein
  • Interferon Regulatory Factors
  • NOTCH2 protein, human
  • Neoplasm Proteins
  • Receptor, Notch2
  • Receptors, Antigen, B-Cell
  • interferon regulatory factor-4
  • interferon regulatory factor-8
  • Tretinoin