Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling by array of human multiple myeloma cells and cell lines; bone marrow stromal cells, bone marrow CD3 cells, bone marrow CD14 cells, bone marrow CD15 cells and in vitro generated osteoclastic cells from myeloma patients; and normal plasmablasts and normal memory B cells.

ABSTRACT: Multiple Myeloma primary myeloma cells of 131 patients, 10 human myeloma cell lines, bone marrow stromal cells of 5 myeloma patients, bone marrow CD3 cells of 5 myeloma patients, bone marrow CD14 cells of 5 myeloma patients, bone marrow CD15 cells of 5 myeloma patients, in vitro generated osteoclastic cells of 7 myeloma patients, 7 normal plasmablasts and 6 normal memory B cells.

ORGANISM(S): Homo sapiens

DISEASE(S): multiple myeloma

SUBMITTER: Jerome Moreaux

PROVIDER: E-TABM-937 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Conventional anti-cancer drug screening is typically performed in the absence of accessory cells (e.g. stromal cells) of the tumor microenvironment, which can profoundly alter anti-tumor drug activity. To address this major limitation, we have developed assays (e.g. the tumor cell-specific in vitro bioluminescence imaging (CS-BLI) assay) to selectively quantify tumor cell viability, in presence vs. absence of non-malignant stromal cells or drug treatment. These assays have allowed us to identify that neoplastic cells from diverse malignancies exhibit stroma-induced resistance to different anti-tumor agents. In this analysis, we evaluated the molecular changes triggered in myeloma cells by their in vitro interaction with stromal cells. The transcriptional profile of 3 human multiple myeloma (MM) cell lines (MM.1S, MM.1R, INA-6) co-cultured with stromal cells vs. when cultured alone was characterized by oligonucleotide microarray analysis, using the human U133 plus 2.0 Affymetrix GeneChip. Three human multiple myeloma (MM) cell lines (MM.1S, MM.1R, INA-6) stably expressing green fluorescent protein (GFP) were cultured in vitro for 24hrs either alone or in the presence of the human bone marrow stromal cell line HS-5. Fluorescence activated cell sorting was used to separate the GFP+ MM cells from GFP- stromal cells. The gene expression profiles of MM cells after their co-culture with stromal cells were compared with the profiles of MM cells cultured alone, according to previously described protocols for total RNA extraction and purification; cDNA synthesis; production of biotin-labeled cRNA; hybridization of cRNA with U133plus2.0 Affymetrix gene chips; and scanning of image output files. Scanned image output files were analyzed using DNA-Chip Analyzer (dChip) (www.dchip.org), including conversion to DCP files, normalization and modeling, and averaging of duplicate chips according to standard parameters recommended by the software. For each cell line, the gene expression profile in the presence of stomal cells was compared to the profile of the same cell line cultured in the absence of stromal cells. Technical replicates for each condition were analyzed (total of 12 gene expression profiles).

Project description:Natural Killer cells (NK), a major constituent of innate immune system, have the ability to kill the transformed and infected cells without prior sensitization; can be put to immunotherapeutic use against various malignancies. NK cells discriminate between normal cells and transformed cells via a balance of inhibitory and activating signals induced by interactions between NK cell receptors and target cell ligands. Present study investigates whether expansion of NK cells could augment their anti-myeloma (MM) activity. For NK cell expansion, peripheral blood mononuclear cells from healthy donors and myeloma patients were co-cultured with irradiated K562 cells transfected with 4-1BBL and membrane-bound IL15 (K562-mb15-41BBL). A genome-wide profiling approach was performed to identify gene expression signature in expanded NK (ENK) cells and non-expanded NK cells isolated from healthy donors and myeloma patients. A specific set of genes involved in proliferation, migration, adhesion, cytotoxicity, and activation were up regulated post expansion, also confirmed by flow cytometry. Exp-NK cells killed both allogeneic and autologous primary MM cells more avidly than non-exp-NK cells in vitro. Multiple receptors, particularly NKG2D, natural cytotoxicity receptors, and DNAM-1 contributed to target lysis, via a perforin mediated mechanism. In summary, vigorous expansion and high anti-MM activity both in vitro and in vivo provide the rationale for testing exp-NK cells in a clinical trial for high risk MM. Differential gene expression profile in expanded natural killer (ENK) cells and non-expanded natural killer (NK) cells from healthy donors and myeloma patients Eight healthy donor and eight myeloma patients were used in the study. Non-expanded natural killer (NK) cells were isolated from PBMCs of healthy donors and myeloma patients. Expanded natural killer (ENK) cells were generated from same set of samples as mentioned in expansion protocol. All ENK and NK cells were used for gene expression profiling.

Dataset Information

Transcription profiling by array of human multiple myeloma cells and cell lines; bone marrow stromal cells, bone marrow CD3 cells, bone marrow CD14 cells, bone marrow CD15 cells and in vitro generated osteoclastic cells from myeloma patients; and normal plasmablasts and normal memory B cells.

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