Abstract A25: Evolving functional heterogeneity in B-acute lymphoblastic leukemia

Abstract

Abstract Despite high survival rates for children with acute lymphoblastic leukemia (ALL), only 40% of adult patients will achieve long-term disease-free survival, and relapses in both pediatric and adult ALL are often fatal. B-ALL leukemic blasts exhibit considerable subclonal genomic diversity. In 50% of patients, the clones present at relapse are not the dominant clone at diagnosis, but have evolved from a minor or ancestral clone. B-ALL subclonal diversity also exists in leukemia-initiating cells (L-IC) that are functionally capable of initiating xenografts. In order to investigate the clonal evolution during disease progression and link genetic diversity with functional characteristics, including differentiation, migratory properties, L-IC frequency and therapy resistance, we performed in depth genomic and functional analysis of 14 paired diagnosis/relapse samples from adult and pediatric B-ALL patients of varying cytogenetics. Time to relapse ranged from 6 to 97 months. Patient samples were subjected to whole exome/genome sequencing, SNP analysis and RNA sequencing. Diagnosis-specific, relapse-specific and shared variants at both clonal and subclonal frequencies were identified. Limiting dilution analysis by transplantation of CD19+ leukemic blasts into immune-deficient mice (xenografts) identified no significant trend in enrichment in L-IC frequency between paired patient samples with a median frequency of 1 in 2691. Despite similar frequencies of L-IC, functional differences within identically sourced patient xenografts were observed, including increased leukemic dissemination of relapse cells to the spleen and/or central nervous system, differences in engraftment levels and differences in immunophenotypes. Copy number analysis and ongoing variant sequencing of the xenografts, in comparison to the patient sample from which they were derived, has uncovered clonal variation and the outgrowth of ‘relapse-like’ subclones in xenografts transplanted with the diagnostic sample. Interrogation of the therapeutic response of these subclones in secondary xenografts displayed evidence of resistance to standard chemotherapeutic agents (vincristine and L-asparaginase). Therefore we have shown evidence that relapse subclones/ ancestral clones present at diagnosis possess functional advantages over other diagnostic clones. Overall, this work will provide further understanding of the heterogeneity identified in B-ALL and how it contributes to lymphoid leukemogenesis, therapy failure, and disease recurrence. Citation Format: Stephanie M. Dobson, Robert Vanner, Esmé Waanders, Jessica McLeod, Olga I. Gan, Zhaohui Gu, Debbie Payne-Turner, Xiaotu Ma, Yiping Fan, Pankaj Gupta, Michael Rusch, John Easton, Cynthia J. Guidos, Jayne S. Danska, Jinghui Zhang, Mark D. Minden, Charles G. Mullighan, John E. Dick. Evolving functional heterogeneity in B-acute lymphoblastic leukemia. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A25.