Tumor cell-specific bioluminescence platform to identify stroma-induced changes to anticancer drug activity

Abstract

Conventional anti-cancer drug screening is typically performed in the absence of accessory cells of the tumor microenvironment, which can profoundly alter anti-tumor drug activity. To address this major limitation, we developed the tumor cell-specific in vitro bioluminescence imaging (CS-BLI) assay. Tumor cells (e.g. myeloma, leukemia and solid tumors) stably expressing luciferase are co-cultured with non-malignant accessory cells (e.g. stromal cells) for selective quantification of tumor cell viability, in presence vs. absence of stromal cells or drug treatment. CS-BLI is high-throughput scalable and identifies stroma-induced chemoresistance in diverse malignancies, including imatinib-resistance in leukemic cells. A stromal-induced signature in tumor cells correlates with adverse clinical prognosis and includes signatures for activated Akt, Ras, NF-κB, HIF-1α, myc, hTERT, and IRF4; signatures for biological aggressiveness and for self-renewal. Unlike conventional screening, CS-BLI can also identify agents with increased activity against tumor cells interacting with stroma. One such compound, reversine, exhibits more potent activity in an orthotopic model of diffuse myeloma bone lesions than in conventional subcutaneous xenografts. Use of CS-BLI, therefore, enables refined screening of candidate anti-cancer agents to enrich preclinical pipelines with potential therapeutics that overcome stroma-mediated drug resistance and can act in a synthetic lethal manner in the context of tumor-stromal interactions.

Preferential killing of p53-deficient cancer cells by reversine

“…35,[41][42][43][44][45] These findings indicate that reversine exerts relatively broad anticancer effects. Here, we show that low-dose reversine, 0.5 μM, a concentration at which reversine selectively inhibits MPS1, fails to preferentially kill TP53 -/-HCT 116 colon carcinoma cells.…”

New Insights into Therapeutic Targets in Myeloma

“…For example, up-regulation of the ubiquitin proteasome cascade at both a gene transcript and activity level in tumor cells is triggered by MM-BM binding, 17 making it an even better target in this context for bortezomib, which is able to overcome cell adhesion-mediated drug resistance to conventional therapy. These model systems allow for defining the impact of agents on the tumor versus the microenvironment.…”

The anti‐cancer agent SU4312 unexpectedly protects against MPP+‐induced neurotoxicity via selective and direct inhibition of neuronal NOS

“…SU4312 competes with ATP for binding to VEGFR-2 and is able to completely block VEGF signalling in a noncompetitive manner (Sun et al, 1998). SU4312 specifically inhibited VEGF-dependent angiogenesis without damaging normal cells (Tran et al, 2007;Miki et al, 2010) and also reduced the proliferation of multiple myeloma and leukaemia tumour cells in vitro (McMillin et al, 2010). It is suggested that the anticancer activity of SU4312 is achieved through direct inhibition of the proliferation of cancer cells and indirect suppression of angiogenesis.…”