LEUKEMIC NICHE MODELING (AML)

Preclinical development

Targeted therapy
Chemotherapy

Preclinical research (TRL 4-5)

Tissue niche model by co-culture in physioxia (physiological oxygen concentration) for the selection and validation of drug candidates for the treatment of acute myeloid leukemia (AML).

The CellOxia cell culture platform offers equipment (XVivo Biosphérix) enabling studies to be carried out under controlled atmospheric conditions, to reproduce in vitro and ex vivo the physiological or pathophysiological parameters existing within the tissue niche, as in the case of leukemic bone marrow.

Description

Scope of research activities

Preclinical research using an in vitro and ex vivo leukemia niche model for the selection and validation of new molecules for the treatment of AML:

Screening
Demonstration of mechanism of action
Efficacy evaluation
In vivo validation in PDX AML

Conduct of studies

Steps:

Study design after definition of the question raised and the services required
Proposal of specifications according to the service required (service or simple use of equipment)
Implementation of services and experiments
Conducts of experiments and analyses of results
Study report for services or experimenters' support

Research infrastructure

Experimental and analysis platforms:

XVivo model X1 (Biosphérix)
P2 culture laboratory
PAULA intelligent cell imaging system (Leica)
Nikon Ti inverted microscope with phase-contrast and fluorescence imaging

Cell models :

Primary cells from AML patients (leukemic blasts)
Mesenchymal or adipocytic cells
Human AML cell lines and human and murine stromal cells

Technical personnel:

Specialized engineer for design and implementation of hypoxia studies

Quality assurance

The CellOxia platform is part of the research support unit (UAR) CNRS 3427/US005 Inserm comprising 11 platforms. Created in 2016 and accredited by Siric Brio, it joined the UAR in 2021. To date, it has 12 academic laboratories and two companies that have or are still using its services and equipment.

Specifications

The platform

A set of in vitro tests, reproducing the pathophysiological conditions of the leukemic niche, for selection and validation of drug candidates for the treatment of AML.

The studies

Screening of drug candidates:

Proliferation and apoptosis assays +/- drugs in hypoxia
Dose-response of agents to different oxygen concentrations
Molecule screening

Evaluation of anti-leukemic activity of drug candidates:

Test in vitro (cell lines), ex vivo (primary cells) or in vivo (xenograft with response and survival monitoring)

Mechanism of action of candidate's anti-leukemic activity:

Analysis of signaling and key points using appropriate techniques (Western-blot, cytometry, PCR-Q, RNASeq)
Modeling by generation of modified cell lines

Examples of partnerships

Search for genes involved in Giltritinib resistance in AML - To identify genes involved in Giltritinib resistance, a pan-genomic CrispR screen was performed under niche conditions (coculture and 3% oxygen).

Partner: Astellas Pharmaceutical

Search for resistance genes to therapeutic combinations in AML - In order to identify the genes involved in resistance to the various combinations used to date in AML (Ara-C, (-aza, Giltéritinib, vénétoclax), a CrispR pan-genomic screening was performed under niche conditions (coculture and 3% oxygen) with these agents alone or in combination.

Partner: Bristol Meyrs Squibb