Long-term in vitro maintenance of clonal abundance and leukaemia-initiating potential in acute lymphoblastic leukaemia

Abstract

Lack of suitable in vitro culture conditions for primary acute lymphoblastic leukaemia (ALL) cells severely impairs their experimental accessibility and the testing of new drugs on cell material reflecting clonal heterogeneity in patients. We show that Nestin-positive human mesenchymal stem cells (MSCs) support expansion of a range of biologically and clinically distinct patient-derived ALL samples. Adherent ALL cells showed an increased accumulation in the S phase of the cell cycle and diminished apoptosis when compared with cells in the suspension fraction. Moreover, surface expression of adhesion molecules CD34, CDH2 and CD10 increased several fold. Approximately 20% of the ALL cells were in G0 phase of the cell cycle, suggesting that MSCs may support quiescent ALL cells. Cellular barcoding demonstrated long-term preservation of clonal abundance. Expansion of ALL cells for >3 months compromised neither feeder dependence nor cancer initiating ability as judged by their engraftment potential in immunocompromised mice. Finally, we demonstrate the suitability of this co-culture approach for the investigation of drug combinations with luciferase-expressing primograft ALL cells. Taken together, we have developed a preclinical platform with patient-derived material that will facilitate the development of clinically effective combination therapies for ALL.

Pro-inflammatory-Related Loss of CXCL12 Niche Promotes Acute Lymphoblastic Leukemic Progression at the Expense of Normal Lymphopoiesis

“…The cell–cell intercommunication as key factor for tumor progression has became a research priority, and increasing evidence of leukemic blasts supported by Nestin + MSC indicates their ability for long-term maintenance at the disease onset (47, 48). MSC are part of the complex tissue network of the BM microenvironment (42) that establishes an essential niche for hematopoiesis (49).…”

Targeting the thioredoxin system as a novel strategy against B‐cell acute lymphoblastic leukemia

“…Isolated BM‐MSC were cultured according to the protocol described in Pal et al . (). The identity and quality of the BM‐MSC was confirmed by the assessment of the positive (CD73, CD105, CD90) and negative (CD45, CD34, CD19) markers by flow cytometry as well as by their potential to differentiate into adipocytes, osteoblasts, and chondroblasts.…”

Stability of patient-specific features of altered DNA replication timing in xenografts of primary human acute lymphoblastic leukemia

“…In addition, conditions that support or stimulate ALL cell survival and proliferation ex vivo are generally lacking or poorly understood, although some progress in this regard has been recently reported (Pal et al, 2016). Genetically engineered mouse models of human leukemia offer an experimental alternative, but these face the same cell growth problems and rarely recapitulate the genetic complexity associated with the clinical examples of the human leukemias they are intended to model (Beer and Eaves, 2015).…”