IN VITRO EVALUATION OF THE EFFECTS OF ANTILEUKEMIC DRUGS IN A MODELED LEUKEMIC NICHE WITH OPTIMIZED BONE MARROW MICROENVIRONMENT (MSCS, HYPOXIA, PRESSURE) (MPN/ALL/AML/MDS)

Preclinical development

Targeted therapy
Precision medicine
Chemotherapy

Preclinical research (TRL 4-5)

In vitro models for assessing the functional impact of antileukemic drugs in a modeled leukemic niche with optimized bone marrow microenvironment (mesenchymal stromal cells, hypoxia, intramedullary pressure).

These models enable us to assess the impact of drug candidates on leukemia cells within the bone marrow, particularly with regard to oxidative and energetic niche metabolism in relation to chemoresistance and pharmaco-modulation.

Description

Scope of research activities

Preclinical research using various in vitro models to test the action of metabolic drugs on the medullary niche:

Assessment of toxicity

Conduct of studies

Steps:

Analysis of innovative product development strategy
Study design based on scientific data and mechanistic hypotheses
Drafting of study plans
Analysis and communication of results

Research infrastructure

Experimental and analysis platforms:

Hypoxia and intramedullary pressure modeling stations (Avatar®)
Flow cytometry: FACS (BD Melody®)
Real-time metabolic analysis (Seahorse®, Omnilog®)
Small animal irradiator (Faxitron®)
Single cell analysis systems (including Chromium Controller 10X Genomics)
Genomic, transcriptomic, metabolomic and lipidomic analyses

Models:

Tumor niche models with primary MSCs and cell lines (MS5, HS27a, HS5)
Primary patient cells
Leukemia cell lines (over 20 available)
Murine leukemias (FLA2, FLB1)

Technical personnel:

4 PhD engineers
3 technicians

Specifications

The platform

Set of preclinical studies using in vitro leukemia models for functional evaluation of the impact of metabolic drugs on the tumor environment.

The studies

Assessment of drug candidate toxicity:

Analysis of cell cycle, apoptosis and DNA breaks
Oxidative (ROS quantification by CMF, antioxidograms, NRF2 pathway, MAPK signaling) and energetic (glycolysis, oxidative phosphorylation) metabolism analyses
Transcriptomic (RNAseq, scRNAseq, RT-qPCR), genomic (NGS mutational profiles) and epigenomic (DNA methylation) analyses
Functional analyses (hematopoietic progenitor cultures, LTC-IC in limiting dilution, MSC differentiation).
Mouse models of stem cell leukemias (FLA2 & FLB1 leukemias / Cf. Herault O et al, J Exp Med 2012, 209:895-901)