Human Multiple Myeloma Cell lines & Molecular Heterogeneity

Human Multiple Myeloma Cell lines & Molecular Heterogeneity 2016-11-04T17:47:51+00:00

Our cellular models

The study of biology of multiple myeloma cells is not easy because plasma cells are in a bone marrow niche and interactions with the microenvironnement are essential for their proliferation and survival.  This is even more difficult for normal plasma cells because they are rare and difficult to harvest; they represent approximately 0.5% of mononuclear cells.

It is from this perspective that the team has developed different unique models to study the biology of the multiple myeloma:

HMCLs (Human Multiple Cell Lines) : 

We have developed a unique large collection of myeloma cell lines (n=40) with 20 cell lines those are dependent of the adding of exogene IL-6 for the growing and survival. These cell lines are representative of the molecular heterogeneity described in patients.

We are characterizing the response of our cell lines collection to 20 drugs including alkylating agents, proteasome inhibitors, IMiDs, corticoid, antimetabolites, NFkappaB inhibitors, CDK inhibitors, aurora kinase inhibitors, epigenetic-targeted treatments (HDACi, BRDi, HMTi…), DNA repair inhibitors and PI3-kinase inhibitors as previously described.

We also derived drug resistant myeloma cell lines (dexamethasone, bortezomib, IMiDs, melphalan, HDACi, …) to explore the development of the myeloma cells resistance of different drug.

The panel of cell lines is also representative of the heterogeneity of patients in the response to the treatment.

This myeloma cell line collection and developed resistant-cell lines have characterized by GEP (Gene Expression Profilling) and recently at different molecular levels using RNA-seq, Exome-seq, Whole genome seq, Chip-seq (6 different histone marks), SNP analysis, DNA methylation analysis and miRNA-seq.

HMCLs are an optimal model to access drug efficiency and to study mechanisms of actions that may play a role in cell death, clonogenicity, proliferation and treatment resistances.

Others hematological cancer cell lines :

In additions of multiple myeloma cell lines, we have 20 diffuse large B-cell lymphoma human cell lines and some human myeloid leukemia cell lines.  These cell lines have used to evaluate effects of different molecule and to validate our patented “DNA repair” risk score build in DLBCL.

In Vitro model of differentiation of memory B cells into plasmablasts and mature plasma cells :

This model has developed by the research team of Jerome Moreaux and allows obtaining CD138+ and immunoglobulin secreting plasma cells starting from memory B cells in a 3-steps and 10 days culture. For each step, the detailed phenotypic characterization of B memory cells and in vitro-generated activated B cells, pre-plasmablasts, plasmablasts and plasma cells had shown by multicolor-cytometry. The different cells of the model were also characterized by gene expression profiling and by next generation sequencing.

Recently, our scientific team has used the same approaches to develop an in vitro model of differentiation of memory B cells into long-lived plasma cells (LLPCs) surviving for at least 4 months in vitro. These LLPCs had produced immunoglobulins continuously, are out of the cell cycle and express highly transcription factors and surface markers of plasma cells.

This original model is perfectly designed to offer the effect of graded concentration of a drug on the ability to generate in vitro activated B cells, plasmablasts and mature plasma cells, on the perturbation of genomic stability of cells but also to explore the development of the cells resistance of different drugs.

Model of chemogram :

We have taken advantage of our expertise in the understanding of MM cells and bone marrow microenvironment interactions and have develop an in vitro coculture model allowing primary MM cells culture in a supportive environment with the major growth factors cell to cell communication signals (in house coculture model – not divulgated). Starting from bone marrow sample, this automatizing system allows testing in vitro primary myeloma cell drug response to 5 – 10 molecules.

Our goal will be to create a new tool to quickly test the sensitivity/resistance of MM cells from patients to new-targeted molecules and improving treatment strategies to develop precision medicine in multiple myeloma.