Project description:Multiple myeloma (MM) is a not curable B-cell neoplasia characterised by the accumulation of tumor plasma cells within the bone marrow. As a consequence, 80% of patients develop bone osteolytic lesions that remain even after complete remission. We and others had demonstrated that bone marrow derived Mesenchymal Stromal Cells (MSC) are abnormal in MM and thus could not be used for an autologous treatment to repare bone damages. Adipose Stromal Cells (ASC) represent an interesting alternative to MSC for cellular therapy. Thus, in this paper we asked if they could be a good candidate in repairing MM bone lesions. For the first time, we present a transcriptomic, phenotypic and functional comparison between ASC from MM patients and healthy donors (HD) relying on their autologous MSC counterpart. In contrast to MM MSC, MM ASC do not exhibit major abnormalities. However, the changes observed in MM ASC and the supportive property of ASC on MM cells question their putative and safety uses at autologous or allogenic level.
Project description:Multiple myeloma (MM) is an incurable B cell neoplasia characterized by the accumulation of tumor plasma cells within the bone marrow (BM). As a consequence, bone osteolytic lesions develop in 80% of patients and remain even after complete disease remission. We and others had demonstrated that BM-derived mesenchymal stromal cells (MSCs) are abnormal in MM and thus cannot be used for autologous treatment to repair bone damage. Adipose stromal cells (ASCs) represent an interesting alternative to MSCs for cellular therapy. Thus, in this study, we wondered whether they could be a good candidate in repairing MM bone lesions. For the first time, we present a transcriptomic, phenotypic, and functional comparison of ASCs from MM patients and healthy donors (HDs) relying on their autologous MSC counterparts. In contrast to MM MSCs, MM ASCs did not exhibit major abnormalities. However, the changes observed in MM ASCs and the supportive property of ASCs on MM cells question their putative and safety uses at an autologous or allogenic level.
Project description:It is now well established that bone marrow (BM) constitutes a microenvironment required for differentiation. Bone marrow mesenchymal stromal cells (BM-MSCs) strongly support MM cell growth, by producing a high level of Interleukin-6 (IL-6), a major MM cell growth factor. BM-MSCs also support osteoclastogenesis and angiogenesis. Previous studies have suggested that the direct (VLA-4, VCAM-1, CD44, VLA-5, LFA-1, syndecan-1,M-bM-^@M-&) and indirect interactions (soluble factors) between MM plasma cells and BM-MSCs result in constitutive abnormalities in BM-MSCs. In particular, MM BM-MSCs express less CD106 and fibronectin and more DKK1, IL-1M-NM-2 and TNF-M-NM-1 as compared with normal BM-MSCs. In order to gain a global view of the differences between BM-MSCs from MM patients and healthy donors, we used gene expression profiling to identify genes associated to the transformation of MM BM-MSCs. BM-MSCs were isolated from 3 healthy donors and 4 untreated multiple myeloma patients. Total RNA from BM-MSCs was exctracted and hybridyzed on Affymetrix GeneChipM-BM-. Human Genome U133 Plus 2.0 Array. Amplification, hybridization and scanning were done according to standard Affymetrix protocols (www.affymetrix.com). CEL files were normalized with RMA method.
Project description:Multiple myeloma is hematologic malignancies result from clonal proliferation of plasma cells. Recently, increasing evidence supports the hypothesis that microenvironment cells play important roles in the proliferation, survival, and drug resistance of clonal plasma cells. The aim of this study is to culture stromal cells from bone marrow aspirates of patients with multiple myeloma, and to investigate expression profiles of bone marrow stromal cells and their relationships with the clinical characteristics of patients. RNA was extracted cultured bone marrow stromal cells from 15 patients with plasma cell neoplasms, and bone marrow stromal cells from 13 control patients with 9 B-cell lymphoma patients with no evidence of BM involvement and 4 patients with mild-to-moderate cytopenia without evidence of hematologic malignancies
Project description:Neutrophils are the most abundant nucleated cell type in the bone marrow. A pro-tumor bias in this cell type may have implications for bone-marrow residing malignancies, such as multiple myeloma. Here, we generated single cell transcriptomic overviews of the entire myeloid compartment, including the entire neutrophilic lineage, of the bone marrow of 6 newly diagnosed myeloma patients, 5 treated myeloma patients and 4 non-cancer controls. We find dat mature neutrophils in myeloma patients, both newly diagnosed and treated, have an activated and pro-inflammatory phenotype, accompanied by increased transcription of pro-inflammatory cytokines, such as IL-1β, and myeloma cell survival factors, such as BCMA-ligand BAFF/TNFSF13B. Moreover, inflammatory stromal cells can activate naive neutrophils to acquire an inflammatory phenotype as is seen in patients. Previously, we have shown that inflammatory stromal cells characterized the bone marrow of newly diagnosed myeloma patients. Here, we generate single cell RNA sequencing dataset of non-hematopoietic bone marrow cells of patients after induction treatment, high-dose melphalan, stem cell transplantation and consolidation treatment. We show that this intensive treatment reduced, but did not normalize, stromal inflammation.
Project description:Multiple myeloma is hematologic malignancies result from clonal proliferation of plasma cells. Recently, increasing evidence supports the hypothesis that microenvironment cells play important roles in the proliferation, survival, and drug resistance of clonal plasma cells. The aim of this study is to culture stromal cells from bone marrow aspirates of patients with multiple myeloma, and to investigate expression profiles of bone marrow stromal cells and their relationships with the clinical characteristics of patients.
Project description:Differential gene expression analysis of three multiple myeloma cell lines without stromal co-culture (control) with stromal co-culture (condition) after 72 hours of exposure.
Project description:Differential chromatin accessibility analysis of three multiple myeloma cell lines without stromal co-culture (control) with stromal co-culture (condition) after 72 hours of exposure.
Project description:An experiment was performed in order to compare directly profiles of stromal vascular fraction of adipose tissue, testis and epidydimis, in order to rule out a possible contamination of the stromal vascular fraction of adipose tissue (SVF) by adjacent tissues (i.e. epididymis and testis). This led to the identification of a list of 2358 SVF-specific probes, which was subsequently used for further investigations. 3 dye-swap pairs, comparing 3 distinct samples of total RNA prepared from: (1) stromal vascular fraction of adipose tissue, (2) testis and (3) epidydimis.
Project description:Multiple myeloma (MM) progression is linked to chronic NF-κB activation in myeloma cells. However, the identity and source of autocrine/paracrine signals driving NF-κB activation and the role of the 3D microenvironment have been scarcely investigated both in vivo and in vitro. To investigate them, we knocked-in the Venus (YFP) ORF in the NF-κB p65 gene in both MM and stromal cells. Surprisingly, a large fraction of p65-YFP MM cells engrafted in mouse bone marrow showed overall low levels of NF-κB activation whereas a small fraction was highly activated. To understand these in vivo data, we investigated NF-κB dynamics in MM and stromal cells, both alone and in co-culture. In vitro experiments exploiting microfluidics, bioreactor and microchip cell cultures highlighted crosstalk between the myeloma and stromal components that leads to mild basal activation. In contrast, we found that high-density cultures within 3D scaffolds dampen NF-κB activation in MM and stromal cells, both in basal and inflammatory conditions. It has been recently hypothesized that IL1β, and the inflammatory ME, shape the overall activity of ME components and promote the transition of Mesenchimal Stromal Cells (MSCs) toward an inflammatory NF-κB driven transcriptional phenotype (iMSCs). We tested this hypothesis in our system and found that IL1β strongly activates NF-κB in stromal but not in myeloma cells. In addition, secreted molecules from IL1β-stimulated MSCs strongly activate NF-κB only in a small fraction of MM cells. We propose that the balance between activating stimuli from iMSCs and dampening feedbacks from the 3D ME, maintains a mild NF-κB activation in myeloma cells in the patients’ BM to avoid exceedingly harmful responses.