PRECLINICAL MODELS FOR THE DEVELOPMENT OF NEW TREATMENTS FOR MYELOPROLIFERATIVE NEOPLASIA

Preclinical development

MPN
MDS
LMMJ

Targeted therapy
Chemotherapy

Preclinical research (TRL 4-5)

In vitro and in vivo models of myeloproliferative neoplasia for preclinical testing to select and validate drug candidates.

Our team uses in vitro models such as cell lines carrying isogenic MPN initiator mutations. This enables us to test the impact of a mutation with the advantages and disadvantages of the cell lines.

We also use samples from patients, usually blood samples, as diseased cells are circulating in MPN. These cells are studied in liquid culture or clonogenic assays, with or without CD34 sorting. The advantage is that we can compare the fate of cells directly derived from patients, carrying the relevant abnormalities. In the various tests, we can compare the fate and drug sensitivity of diseased cells and residual normal cells from the same patient.

Description

Scope of research activities

Preclinical research using different specific models of myeloid leukemia or MPN to test drug candidates:

Demonstration of mechanism of action
Evaluation of efficacy
Evaluation of toxicity

Conduct of studies

Steps:

Design and drafting of study plan
Organization, implementation and conduct of studies
Analysis and communication of results

Research infrastructure

Experimental and analytical platforms:

Flow cytometry
Genomics
Cell culture

Models:

Murine leukemia models
Primary cells from MPN patients: blood or bone marrow hematopoietic cells, stromal cells.
Leukemia cell lines secondary to MPN

Specifications

The platform

Preclinical studies using in vitro and in vivo MPN models.

The studies

In vitro studies are carried out either in liquid medium or in semi-solid medium (clonogenic culture) on total mononuclear cells or cells sorted on CD34. A mouse model carrying the MPN-initiating JAK2 mutation is also used in this study.

Evaluation of anti-leukemic activity of drug candidates:

Cell survival or apoptosis rate
Number of colonies and ratio to untreated
% of colonies remaining after treatment and carrying the initiating mutation
Allelic load of the initiating mutation and additional mutations after culture with the drug candidate
Mouse model: monitoring of blood parameters and survival of mice

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

Study of signaling pathways activated by initiating mutations

Assessment of drug candidate toxicity:

Same items as for anti-leukemic activity, but on the non-mutated population of patient samples
Survival, apoptosis, clonogenicity on samples from healthy subjects obtained from EFS

Examples of partnerships

Study of the efficacy of Ropeg Interferon alpha on JAK2-mutated MPN cells - Test of antiproliferative action on JAK2-mutated cell lines and inhibition of clonogenic sprouting of JAK2-mutated hematopoietic progenitors. Comparison with the reference form of pegylated interferon alpha.

Partner: AOP Orphan

Testing the efficacy of an MDM2 inhibitor in MPN - In vitro and in vivo tests in JAK2-mutated MPN models of the efficacy of the MDM2 inhibitor.

Partner: KARTOS