Project description:We performed whole genome profiling in order to determine the landscape of genetic alterations assoicated with a subset of CLL that is characterized by deletions in 17p The number of copy number alterations predicted shorter time to treatment among patients untreated at sampling. Chromosome 3p, 4p, and 9p were frequently deleted in del(17p) CLL and strongly associated with shorter OS. We conclude that del(17p) has a unique genomic profile characterized typically by TP53 mutation with novel CNAs and novel drivers, with increasing genomic complexity of any type associated with worse overall survival.
Project description:Malignant melanoma is a highly metastatic disease disseminating to several distant sites. This potential is also of great clinical impact for patient survival and therapeutic success. Knowledge about melanoma genomics is mainly based on lymphatic or skin metastases derived data, whereas data from distant sites is rather limited. Therefore, an autopsy-based visceral metastasis biobank was established and an array-based CNV analysis was performed focusing primarily on major organs (brain, lung and liver) on a total of 38 samples (10 primary and 28 organ metastasse). A unique picture emerged about organ specific CNV type distributions or gene alterations including the frequent loss of DNA damage error genes in brain metastases, the presence of HGF/MET autocrine loop in brain and lung metastases, the traces of immunogenic mimicry exclusive for lung metastases or the correlation of BRAF copy number and mutant allele frequency especially in lung metastases. All these above phenomena have a great influence on therapy efficacy or resistance.
Project description:Acute myeloid leukemia (AML) is a clonal hematopoietic malignancy, characterized by expansion of immature leukemic blasts in the bone marrow. In AML, specific tyrosine kinases have been implicated in leukemogenesis, and are associated with poor treatment outcome. However, targeted therapy using kinase inhibitors (KIs) has had limited success, and may be improved by proper patient selection. We performed phosphotyrosine (pY) based, label-free phosphoproteomics to identify hyperphosphorylated, active kinases in AML cell lines as targets and predictive biomarkers to select KIs for treatment. We identified 3605 class I phosphorylation sites in 16 AML cell lines (EOL-1, KG-1a, MM6, KG-1, ME-1, NB-4, Kasumi-3, MV4-11, THP-1, HEL, HL-60, Kasumi-1, Kasumi-6, ML-2, OCI-AML3, MOLM-13) that exhibited large variation in the number and level of phosphopeptides per cell line (241-2764). Ranking analyses successfully pinpointed the hyperactive kinases PDGFRA, FGFR1, KIT, and FLT3 in eight cell lines with a corresponding kinase mutation. Additionally, we identified unexpected drivers in two more cell lines (PDGFRA in Kasumi-3 and FLT3 in MM6) which proved sensitive to specific kinase inhibitors. Six cell lines without a clear receptor tyrosine kinase (RTK) driver showed evidence of MAPK1/3 activation, consistent with the presence of activating RAS mutations. Our data show the potential of pY phosphoproteomics to identify key drivers in AML cells, and the predictive value of the phosphoproteome profiles in TKi selection for targeted treatment.
Project description:Pediatric patients with de novo acute myeloid leukemia. To define genomic architecture, we performed genome-wide copy number abberation analysis in 460 paired diagnostic-remission bone marrow aspirates.
Project description:Neuroblastomas are characterized by recurrent segmental and/or numerical chromosomal abberations such as MYCN-amplification or 11q-deletion. To further elucidate recurrent chromosomal alterations, 16 neuroblastoma cell lines were investigated.
Project description:Development of B-acute lymphoblastic leukemia accompanies with multiple variable mutations. Beside the structural and chromosomal alterations, especially mutations in the regulators of B cell differentiation are common. Around 60% of the B-ALL show deletions of these genes.
Project description:Cell lines. The myeloid cell lines CMK, HEL, HL-60, K-562, KASUMI-1, KG-1, LAMA-84, M-07e, MONO-MAC-1, MV4-11, NB-4, OCI-AML2, OCI-AML5, SIG-M5, THP-1, and UT-7 were obtained from the DSMZ in Braunschweig, Germany (http://www.dsmz.de). The cell line ME-1 was kindly provided by Dr. Ikuya Sakai, Ehime University, Japan. Cell lines were cultured under standard conditions in RPMI 1640 containing 10% FCS, 2 mM L-glutamine, 100 units/ml penicillin and 100 units/ml streptomycin. For subsequent DNA and RNA isolation 1x107 cells of the respective cell lines were washed twice in phosphate-buffered saline, and stored as cell pellets at 800C. Gene expression profiling. Gene expression profiling (GEP) was performed essentially as described using the previously described cDNA microarray platform (Bullinger et al., N Engl J Med 2004;350:1605-1616). A reference experiement design type is where all samples are compared to a common reference. Keywords: reference_design
