Project description:Cancer stem cells (CSCs) that display tumor-initiating properties have recently been identified. We herein identify and characterize CSCs in human uterine carcinosarcoma, a highly aggressive and therapy-resistant gynecologic malignancy, which is considered to be of mesodermal origin. FU-MMT-1, a cell-line, which was established by us (Emoto M, Cancer 1992) from a patient with uterine carcinosarcoma, was evaluated. FU-MMT-1 contained a high population of CD133, CD44, CD90, and CD29 positive cells. Using the magnetic bead cell separation method, we isolated CD133+ cells, which predominantly form spheres in culture. These CD133+ cells form transplantable tumors in vivo. A qRT-PCR analysis of the genes implicated in stem cell maintenance revealed that CD133+ cells express significantly higher levels of OCT4, NANOG, and BMI-1 than CD133M-oM-<M- cells. Moreover, CD133+ cells showed a high expression of PAX2 and WNT4, which are the essential genes in Mullerian duct formation. The tumor derived from CD133+ cells replicated vimentin, ERM-NM-1, ERM-NM-2, and PR expressionsM-cM-^@M-^@of the parent tumor. These findings suggest that CD133+ FU-MMT-1 cells have the characteristics of CSCs and Mullerian mesenchymal progenitors. CD133+ and CD133- population of FU-MMT-1 cells were analyzed by microarray.
Project description:Cancer stem cells (CSCs) that display tumor-initiating properties have recently been identified. We herein identify and characterize CSCs in human uterine carcinosarcoma, a highly aggressive and therapy-resistant gynecologic malignancy, which is considered to be of mesodermal origin. FU-MMT-1, a cell-line, which was established by us (Emoto M, Cancer 1992) from a patient with uterine carcinosarcoma, was evaluated. FU-MMT-1 contained a high population of CD133, CD44, CD90, and CD29 positive cells. Using the magnetic bead cell separation method, we isolated CD133+ cells, which predominantly form spheres in culture. These CD133+ cells form transplantable tumors in vivo. A qRT-PCR analysis of the genes implicated in stem cell maintenance revealed that CD133+ cells express significantly higher levels of OCT4, NANOG, and BMI-1 than CD133- cells. Moreover, CD133+ cells showed a high expression of PAX2 and WNT4, which are the essential genes in Mullerian duct formation. The tumor derived from CD133+ cells replicated vimentin, ERα, ERβ, and PR expressions of the parent tumor. These findings suggest that CD133+ FU-MMT-1 cells have the characteristics of CSCs and Mullerian mesenchymal progenitors.
Project description:When assembling a nephron during development a multipotent stem cell pool becomes restricted as differentiation ensues. A faulty differentiation arrest in this process leads to transformation and initiation of a Wilms' tumor. Mapping these transitions with respective surface markers affords accessibility to specific cell subpopulations. NCAM1 and CD133 have been previously suggested to mark human renal progenitor populations. Herein, using cell sorting, RNA sequencing, in vitro studies with serum-free media and in vivo xenotransplantation we demonstrate a sequential map that links human kidney development and tumorigenesis; In nephrogenesis, NCAM1+CD133- marks SIX2+ multipotent renal stem cells transiting to NCAM1+CD133+ differentiating segment-specific SIX2- epithelial progenitors and NCAM1-CD133+ differentiated nephron cells. In tumorigenesis, NCAM1+CD133- marks SIX2+ blastema that includes the ALDH1+ WT cancer stem/initiating cells, while NCAM1+CD133+ and NCAM1-CD133+ specifying early and late epithelial differentiation, are severely restricted in tumor initiation capacity and tumor self-renewal. Thus, negative selection for CD133 is required for defining NCAM1+ nephron stem cells in normal and malignant nephrogenesis. Human fetal kidney mRNA profiles of 3 cell populations (NCAM1+/CD133-, NCAM+/CD133+, NCAM-/CD133+) were generated by deep sequencing using Illumina HiSeq.
Project description:When assembling a nephron during development a multipotent stem cell pool becomes restricted as differentiation ensues. A faulty differentiation arrest in this process leads to transformation and initiation of a Wilms' tumor. Mapping these transitions with respective surface markers affords accessibility to specific cell subpopulations. NCAM1 and CD133 have been previously suggested to mark human renal progenitor populations. Herein, using cell sorting, RNA sequencing, in vitro studies with serum-free media and in vivo xenotransplantation we demonstrate a sequential map that links human kidney development and tumorigenesis; In nephrogenesis, NCAM1+CD133- marks SIX2+ multipotent renal stem cells transiting to NCAM1+CD133+ differentiating segment-specific SIX2- epithelial progenitors and NCAM1-CD133+ differentiated nephron cells. In tumorigenesis, NCAM1+CD133- marks SIX2+ blastema that includes the ALDH1+ WT cancer stem/initiating cells, while NCAM1+CD133+ and NCAM1-CD133+ specifying early and late epithelial differentiation, are severely restricted in tumor initiation capacity and tumor self-renewal. Thus, negative selection for CD133 is required for defining NCAM1+ nephron stem cells in normal and malignant nephrogenesis.
Project description:In this study, we present a case of parotid gland de novo carcinosarcoma. Salivary gland carcinosarcoma (or true malignant mixed tumor) is a rare biphasic neoplasm, composed of both malignant epithelial and malignant mesenchymal components. It is yet unclear whether these two phenotypes occur by collision of two independent tumors or if they are of clonal origin. To analyze the clonality of the different morphologic tumor components, oligonucleotide microarray-based comparative genomic hybridization (oaCGH) was performed on the carcinoma and the sarcoma entity separately. This technique enables a high-resolution, genome-wide overview of the chromosomal alterations in the distinct tumor elements. Analysis of both fractions showed a high number of DNA copy number changes. Losses were more prevalent than gains (82 and 49, respectively). The carcinomatous element displayed more chromosomal aberrations than the sarcomatous component. Specific amplifications of MUC20 (in mesenchymal element) and BMI-1 (in both elements) loci were observed. Overall homology between the two genomic profiles was 75%. DNA copy number profiles of the epithelial and mesenchymal components in this salivary gland carcinosarcoma displayed extensive overlap, indicating a monoclonal origin. Since losses are shared to a larger extent than gains, they seem to be more essential for initial oncogenic events. Furthermore, specific amplifications of a mucin and a Polycomb group gene imply these proteins in the tumorigenesis of carcinosarcomas.
Project description:Carcinosarcoma of the uterus or ovary is a rare, biphasic tumor comprising epithelial and mesenchymal elements, and exhibits aggressive clinical features. Four molecular subtypes of carcinosarcoma (POLE, MSI, CNH, and CNL) were recently established and shown to be associated with multiple clinicopathological parameters including patient outcomes. Immune microenvironment analyses on CS samples was performed using immune cell profiling with T-cell receptor repertoire assay. Carcinoma and sarcoma elements from CS samples were also assessed separately.
Project description:Cancer stem-like cells are hypothesized to be the major tumor initiating cell (TIC) population of human cutaneous squamous cell carcinoma (hcSCC), but the molecular alterations underpinning their cellular phenotype remain undefined. Here, a small CD133+CD31-CD45-CD61-CD24- (CD133+) cell population enriched for self-renewing and TIC features was isolated by sorting from primary hcSCC tumors. CD133+ cells show enhanced spheroid formation in vitro and tumor generation in vivo. Gene expression profiling comparing CD133+ with CD133- cells revealed that CD133+ cells expressed enriched stem cell-like and cancer-related gene signatures. Eighty differentially expressed stem cell-related genes were identified in CD133+ cells, where the top pathways include Notch, Notch1-mediated NF-κB, and WNT signaling. Overexpression of growth factor receptors, PI3K/mTOR, and STAT pathway genes and inactivation of genes regulating epigenetic modification of chromatin implicated in cancer were also identified. Verification of these gene signatures was conducted with Nanostring in an independent tumor set. Pharmacologic or genetic modulation of Notch1, IKKα, RELA and RELB modulated NF-κB transactivation, the CD133+ population, and phenotype. Immunofluorescent staining confirmed co-localization of CD133+ and IKKα expression in SCC tumor specimens. Our data support the linkage and importance of non-canonical Notch1 and IKK-mediated NF-κB activation in promoting CD133+ population and TIC phenotype in hcSCC.
Project description:The neural stem cell marker CD133 is reported to identify cells within glioblastoma (GBM) that can initiate neurosphere growth and tumor formation, however, instances of CD133- cells exhibiting similar properties have also been reported. Here, we show that some PTEN-deficient GBM tumors produce a series of CD133+ and CD133- self-renewing tumor-initiating cell types and provide evidence that these cell types constitute a lineage hierarchy. Our results show that the capacities for self-renewal and tumor initiation in GBM need not be restricted to a uniform population of stem-like cells, but can be shared by a lineage of self-renewing cell types expressing a range of markers of forebrain lineage. Keywords: Expression and copy number analysis of glioblastomas and neurosphere forming derivative cell lines of same.
Project description:CD133 is a marker of cancer stem cells. DAP5 is a is a translation initiation factor. The goal of the experiment was to characterize the proteomic differences between CD133+/- cells in the WT vs DAP5 depleted background. To this aim, 4 populations of human cells were FACS sorted: shNT_CD133-, shNT CD133+, shDAP5_ CD133-, shDAP5_CD133+. The collected cell pellets were subjected to LC-MS/MS analysis.