Differentiation-dependent EBF1 activity determines <i>CD22</i> transcription and leukemia sensitivity to inotuzumab ozogamicin

Inotuzumab ozogamicin (InO) is an antibody-calicheamicin conjugate with high efficacy in lymphoid malignancies. It targets the B-cell surface protein CD22, which is expressed in most B-cell acute lymphoblastic leukemia (B-ALL) cases, albeit with variable intensity. However, factors governing CD22 expression and thus leukemia sensitivity to InO remain incompletely understood. Using multiomic characterization of 196 human B-ALL samples, coupled with ex vivo InO sensitivity profiling, we showed that early leukemia differentiation arrest at the pre-pro-B stage is associated with resistance to InO. Screening of 1639 transcription factor genes identified early B-cell factor 1 (EBF1) as a key regulator of CD22 expression (false discovery rate of 7.1 × 10-4). When comparing the assay for transposase-accessible chromatin with sequencing profiling results of the most InO-sensitive and -resistant cases (50% lethal concentration <10th vs >90th percentile, n = 18), the binding motif for EBF1 was strikingly enriched in regions with differential open chromatin status (P = 8 × 10-174). CRISPR interference targeting EBF1 binding sites at the CD22 locus led to an ∼50-fold reduction in cell surface CD22 expression and, consequently, an ∼22-fold increase in InO resistance in ALL cell lines. Interestingly, within BCR::ABL1 ALL, we observed intrasubtype heterogeneity linked to EBF1 transcriptional downregulation (P = 1.1 × 10-15) and/or somatic alteration (P = .004), which led to reduced CD22 expression (P = 8.3 × 10-11) and ex vivo and in vivo resistance to InO. Collectively, these findings point to the direct impact of EBF1 on CD22 expression during B-cell development, which, in turn, contributes to interpatient variability in InO response, even within the same subtype of B-ALL.

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Journal
Blood
Published
2025-05-28
DOI
https://doi.org/10.1182/blood.2024028215
Citations
4
Primary Topic
CAR-T cell therapy research
Type
article
Field-Weighted Citation Impact
4.18

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article

Differentiation-dependent EBF1 activity determines CD22 transcription and leukemia sensitivity to inotuzumab ozogamicin

Wendy Stock, Mignon L. Loh, William E. Evans, Megan Walker et al.
4 citations
Blood
CAR-T cell therapy research
4.18
article

Differentiation-dependent EBF1 activity determines CD22 transcription and leukemia sensitivity to inotuzumab ozogamicin

Wendy Stock, Mignon L. Loh, William E. Evans, Megan Walker, Steven Mitchell Kornblau, Ching‐Hon Pui, Marina Konopleva, Wenjian Yang, Carolin S. Escherich, Kelly R. Barnett, Jun Yang, Hiroto Inaba, Elisabeth Paietta, Xin Huang, Mark Robert Litzow, Daniel Savic, Zhenhua Li, Yizhen Li, Satoshi Yoshimura, Jiyang Yu, Elias Jabbour
article en
4 citations

Abstract

Inotuzumab ozogamicin (InO) is an antibody-calicheamicin conjugate with high efficacy in lymphoid malignancies. It targets the B-cell surface protein CD22, which is expressed in most B-cell acute lymphoblastic leukemia (B-ALL) cases, albeit with variable intensity. However, factors governing CD22 expression and thus leukemia sensitivity to InO remain incompletely understood. Using multiomic characterization of 196 human B-ALL samples, coupled with ex vivo InO sensitivity profiling, we showed that early leukemia differentiation arrest at the pre-pro-B stage is associated with resistance to InO. Screening of 1639 transcription factor genes identified early B-cell factor 1 (EBF1) as a key regulator of CD22 expression (false discovery rate of 7.1 × 10-4). When comparing the assay for transposase-accessible chromatin with sequencing profiling results of the most InO-sensitive and -resistant cases (50% lethal concentration <10th vs >90th percentile, n = 18), the binding motif for EBF1 was strikingly enriched in regions with differential open chromatin status (P = 8 × 10-174). CRISPR interference targeting EBF1 binding sites at the CD22 locus led to an ∼50-fold reduction in cell surface CD22 expression and, consequently, an ∼22-fold increase in InO resistance in ALL cell lines. Interestingly, within BCR::ABL1 ALL, we observed intrasubtype heterogeneity linked to EBF1 transcriptional downregulation (P = 1.1 × 10-15) and/or somatic alteration (P = .004), which led to reduced CD22 expression (P = 8.3 × 10-11) and ex vivo and in vivo resistance to InO. Collectively, these findings point to the direct impact of EBF1 on CD22 expression during B-cell development, which, in turn, contributes to interpatient variability in InO response, even within the same subtype of B-ALL.

BloodVol. 146(4)
Albert Einstein College of Medicine (US), St. Jude Children's Research Hospital (US), Mayo Clinic (US), Montefiore Medical Center (US), The University of Texas MD Anderson Cancer Center (US), WinnMed (US), Soochow University (CN), University of Chicago (US), Seattle Children's Hospital (US), Children's Hospital of Suzhou University (CN), Heinrich Heine University Düsseldorf (DE)
Amgen, American College of Radiology Imaging Network, St. Jude Children's Research Hospital, Incyte, ECOG-ACRIN Cancer Research Group, Deutsche Forschungsgemeinschaft, Servier, National Institutes of Health, National Cancer Institute, National Institute of General Medical Sciences
Good health and well-being
Openalex Percentile: Top 6%
CAR-T cell therapy research
4.18
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