Development of a 2D leukaemia-stroma model for analyses of chemo-toxicity protection using a novel bacterial bioluminescent biosensor

Gynn, Liana; May, Jennifer; Anderson, Elizabeth; Robinson, Gareth

Abstract

This study aimed to develop a 2D leukaemia-stroma model of the bone marrow for analysis of stromal-mediated chemo-protection; known to contribute to chemotherapy resistance. The model must facilitate integration of a bioluminescent biosensor HA1 for chemo sensitivity testing and evaluation of clinical resistance. Leukaemic (HL-60) cells were exposed to conditioned media from stromal (HS-5) cells for 24h prior to treatment with 25 µM cytarabine (ara-C) and/or 5 µM daunorubicin (DA); typical induction therapy for AML. Chemo protection facilitated by HS-5 was determined by cytotoxicity assays and validated against active ara-C metabolite generation using the biosensor. Ara-C induced 40.4% cytotoxicity in HL-60 AML cells, which was reduced to 14% when co-cultured with 25% HS-5 conditioned media (p< 0.01). Preliminary investigations suggest this to be comparable to reduced ara-C metabolite chemo-sensitivity of 23% using co-culture inserts (biosensor assay). This is promising indication for development of a model that will assist in the ongoing evaluation of the biosensor’s physiological and clinical relevance. Future work will further optimise these assays, in addition to genotoxicity assessment to define chemo-protection mechanisms. Combining established interaction and toxicity methodologies with this biosensor assay is a novel way of determining drug-specific chemo-protection in tumour micro-environments.

• Presentation Conference Type: Conference Abstract
• Conference Name: 40th Annual Meeting of the United Kingdom Environmental Mutagen Society
• Acceptance Date: May 1, 2017
• Online Publication Date: Feb 8, 2018
• Publication Date: Nov 1, 2017
• Deposit Date: Jan 20, 2020
• Print ISSN: 0267-8357
• Publisher: Oxford University Press (OUP)
• Peer Reviewed: Peer Reviewed
• Volume: 32
• Issue: 6
• Pages: 619
• DOI: https://doi.org/10.1093/mutage/gex033
• Public URL: https://uwe-repository.worktribe.com/output/5190399