To the Novel Paradigm of Proteome-Based Cell Therapy of Tumors: Through Comparative Proteome Mapping of Tumor Stem Cells and Tissue-Specific Stem Cells of Humans

Abstract

We performed proteome mapping (PM), cataloging, and bioinformation analysis of protein lysates of human neural (CD133+) progenitor and stem cells (NPSCs) isolated from the olfactory sheath of a nose, multipotent mesenchymal (CD29+, CD44+, CD73+, CD90+, CD34-) stromal cells (MMSCs) isolated from human bone marrow, and tumor (CD133+) stem cells (TSCs) isolated from the human U87 glioblastoma (GB) cell line. We identified 1,664 proteins in the examined lysates of stem cells (SCs), 1,052 (63.2%) of which are identical in NPSCs and TSCs and 607 proteins (36.47%) of which are identical in MMSCs and TSCs. Other proteins in U87 GB TSCs are oncospecific or carcinogenesis associated. The biological processes, molecular functions, cell localization, and protein signal pathways of the proteins available in all three proteomes were annotated by PubMed (http://www.ncbi.nlm.nih.gov/pubmed/), PANTHER (http://www.pantherdb.org/), GeneOntology (http://www.geneontology.org/), and KEGG (http://www.genome.jp/kegg/) databases. It was shown that gliomaspheres of U87 GB had only 10 intracellular signal transduction pathways (ISTP) that were not modified by the neoplastic process, but only two of them (integrin and focal adhesion pathways) were accessible for regulatory action on gene candidates in the TSC nucleus. Carcinogenesis-free membrane proteins, IPST, and genes expressing proteins of these pathways in U87 GB TSCs can be viewed as main targets for regulatory effects on TSCs. We offer a novel concept of proteome-based complex therapy of tumors. This manuscript is published as part of the International Association of Neurorestoratology (IANR) special issue of Cell Transplantation.

Molecular mechanisms of the effect of TGF-β1 on U87 human glioblastoma cells

“…Tumor cell lysates were extracted and low molecular weight compounds were removed using a previously described method (18).…”

“…Ammonium bicarbonate was used as LC-MS eluent additive (buffer). Trypsin (Thermo Fisher Scientific) was used for trypsinization, as described previously (18). Following completion of this reaction, the samples were concentrated at 30˚С in a Labconco CentriVap centrifugal concentrator to remove all ammonium bicarbonate.…”

“…Tryptic peptides were diluted during the mobile phase (30% acetonitrile, 70% water and 0.1% formic acid; рН 2.7) and divided into 24 fractions using a Dionex UltiMate 3000 chromatograph (Dionex, Sunnyvale, CA, USA) equipped with a fraction collector and MIC-10-CP cation-exchange column (Poros 10S material, 1 mm x 10 cm; Thermo Fisher Scientific, Inc.) (18). The resulting fractions were concentrated at 30˚С in the centrifugal concentrator and re-diluted with 100 µl 0.1% formic acid.…”

“…The analysis of tryptic peptides was performed using a nano-HPLC Dionex Ultimate 3000 and LTQ Orbitrap XL™ mass spectrometer (Thermo Fisher Scientific, Inc.) with a NanoSpray Ionization source (18 The detection sensitivity range, also termed dynamic range, of the mass-spectrometer for the identified proteins was 6-fold (between 4.1x10 8 and 363), which allowed for the detection of proteins with low-copy number, including cofilin 2 and tumor necrosis factor (ligand) superfamily, member 13. Additionally, specific markers of mesenchymal (cluster of differentiation (CD)44, integrin α-5 and β-1) and progenitor stem cells of glioblastoma (aldehyde dehydrogenase 1 family, member A3), and proneural differentiation (tubulin, β3 and nestin) of neural progenitor (nestin) stem cells, were identified.…”

Cancer stem cells and microglia in the processes of glioblastoma multiforme invasive growth

“…One of the possible ways to improve the treatment of glioblastoma is to develop biomedical drugs that are able to control the proliferative and reproductive functions of CSCs. The development of such medication is more successful when using patients' own stem cells (SCs) as a basic material (4–6). …”