PLATFORM FOR SCREENING MOLECULES ABLE OF REPROGRAMMING PRIMARY HUMAN MACROPHAGES (AML/Myelofibrosis/MDS/CMML)

Preclinical development

AML
Myelofibrosis
MDS
CMML

Targeted therapy
Immunotherapy
Alternative therapy

Preclinical research (TRL 4-5)

Analytical platform (MACROTOOLS) for the identification of compounds with the ability to modulate or reprogram macrophage phenotype and function.

The immunomodulatory effect of compounds is evaluated in ex vivo human macrophage models from healthy subjects or patients, using an integrative analysis approach. The platform can test some thirty different compounds every day.

Description

Scope of research activities

Preclinical research using ex vivo models to select macrophage immunomodulating compounds:

Screening
Evaluation of toxicity, reprogramming

Conduct of studies

Steps:

Study strategy analysis
Study design
Drafting of study plans
Organization, implementation and conduct of studies
Analysis and communication of results

Research infrastructure

Experimental and analysis platforms:

Flow cytometry
RT-qPCR
ELISA (Simple PlexTM assays)
Metabolomics: Seahorse, Ysi, Omnilog
T-cell polarization
Phagocytosis tests

Cellular models:

Ex vivo macrophage models: immature macrophages (M0-type macrophages), pro-inflammatory macrophages (M1-type macrophages) and anti-inflammatory macrophages (M2-type macrophages),

Technical personnel:

2 engineers

Specifications

The platform

Ex vivo analysis platform for assessing the effect of compounds on the reprogramming of human macrophages.

The studies

Screening:

Flow cytometry: analysis of the expression of pro- or anti-inflammatory membrane markers (CD80, CD86, HLA-DR, CD163, CD206, CD209, CD200R)
RT-qPCR: analysis of mRNA expression coding for pro- or anti-inflammatory cytokines
ELISA (Simple PlexTM assays): Analysis of the expression of pro- or anti-inflammatory cytokines secreted into the cytokine culture medium
Metabolomics (Seahorse, Ysi, Omnilog): detect and quantify oxidative and glycolytic metabolic processes at the cellular level to assess the metabolic effects of molecules of interest
T-cell polarization: the immunosuppressive capacity of macrophages treated or untreated with molecules of interest is assessed by co-culturing macrophages with naive T cells. The impact of co-culture on T lymphocytes is analyzed by flow cytometry, focusing on the expression of membrane or intracytoplasmic markers specific to Treg, Th1 or Th2 lymphocytes
Phagocytosis tests: the aim of this test is to determine the ability of the molecules of interest to modulate the capacity of macrophages to phagocytose leukemia cells

Toxicity assessment:

The toxicity of molecules of interest is assessed by DAPI labeling, enabling us to evaluate the non-toxic dose to be used on macrophages

Terms

Research services contract

OPALE entity

C3M (UMR-1065)

Contact

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