Supplementary Figure 1: Mouse generation and GSEA enrichment analysis of Sca1-BCR-ABLp190 mice. Supplementary Figure 2: Time course of the percentage of B220+ cells in PB of Sca1-BCR-ABLp190 and Sca1-BCR-ABLp190+Pax5+/- mice. Supplementary figure 3: pB-ALL is transplantable to secondary recipients. Supplementary figure 4: Sca1-BCR-ABLp190+Pax5+/- mice housed in SPF develop similar pB-ALL than Sca1-BCR-ABLp190+Pax5+/- mice housed in CF Supplementary Figure 5: transcriptome analysis of Sca1-BCR-ABLp190+Pax5+/- mice. Supplementary Figure 6: Mouse tumor exome sequencing data identified recurrent Pax5 mutations. Supplementary Figure 7: Polyclonal VDJ recombination events in pre-leukemic B220-positive cells from the BM of Sca1-BCR-ABLp190+Pax5+/- mice Supplementary Figure 8: Altered pathways in leukemic Sca1-BCR-ABLp190+Pax5+/-. Supplementary Figure 9: Quantitative differences of mRNA levels of glycolytic related genes between leukemic Sca1-BCR-ABLp190+Pax5+/-, Sca1-BCR-ABLp190 and wild-type B cells. Supplementary Figure 10: QRT-PCR validation of microarray data. Supplementary Figure 11: Metabolic signature in human pB-ALLs. Supplementary Figure 12: Analysis of ChIP-seq data revealed novel targets of PAX5 in human B lymphocytes. Supplementary Table 1: Percentage and total numbers of hematopoietic progenitor compartments in young Sca1-BCR-ABLp190 and wild type mice. Supplementary Table 3: pB-ALL genotype and phenotype of Sca1-BCR-ABLp190+Pax5+/- and Sca1-BCR-ABLp190 mice.
Funding
German Cancer Aid
Translational Oncology Program
German Jose Carreras Foundation
Hochschule Bonn-Rhein-Sieg
NIH/NCI
Wellcome Trust
Leukemia and Lymphoma Society
Norman and Sadie Lee Foundation
Dr. Ralph and Marian Falk Medical Research Trust
Cancer Research Institute through a Clinic and Laboratory Integration Program
California Institute for Regenerative Medicine (CIRM)
German Children's Cancer Foundation and the Federal Ministry of Education and Research, Bonn, Germany
FEDER
MINECO
Instituto de Salud Carlos III
ISCIII- Plan de Ayudas IBSAL 2015 Proyectos Integrados
Junta de Castilla y León
European Union's Seventh Framework Programme
German Carreras Foundation
Ministerio de Economía y Competitividad
FSE-Conserjería de Educación de la Junta de Castilla y León
Deutsche Forschungsgemeinschaft (DFG) fellowship
ARTICLE ABSTRACT
Preleukemic clones carrying BCR-ABLp190 oncogenic lesions are found in neonatal cord blood, where the majority of preleukemic carriers do not convert into precursor B-cell acute lymphoblastic leukemia (pB-ALL). However, the critical question of how these preleukemic cells transform into pB-ALL remains undefined. Here, we model a BCR-ABLp190 preleukemic state and show that limiting BCR-ABLp190 expression to hematopoietic stem/progenitor cells (HS/PC) in mice (Sca1-BCR-ABLp190) causes pB-ALL at low penetrance, which resembles the human disease. pB-ALL blast cells were BCR-ABL–negative and transcriptionally similar to pro-B/pre-B cells, suggesting disease onset upon reduced Pax5 functionality. Consistent with this, double Sca1-BCR-ABLp190+Pax5+/− mice developed pB-ALL with shorter latencies, 90% incidence, and accumulation of genomic alterations in the remaining wild-type Pax5 allele. Mechanistically, the Pax5-deficient leukemic pro-B cells exhibited a metabolic switch toward increased glucose utilization and energy metabolism. Transcriptome analysis revealed that metabolic genes (IDH1, G6PC3, GAPDH, PGK1, MYC, ENO1, ACO1) were upregulated in Pax5-deficient leukemic cells, and a similar metabolic signature could be observed in human leukemia. Our studies unveil the first in vivo evidence that the combination between Sca1-BCR-ABLp190 and metabolic reprogramming imposed by reduced Pax5 expression is sufficient for pB-ALL development. These findings might help to prevent conversion of BCR-ABLp190 preleukemic cells.Significance: Loss of Pax5 drives metabolic reprogramming, which together with Sca1-restricted BCR-ABL expression enables leukemic transformation. Cancer Res; 78(10); 2669–79. ©2018 AACR.
