46 results on '"Ansén S"'
Search Results
2. Whole-body positron emission tomography using 18F-fluorodeoxyglucose for initial staging of patients with Hodgkin's disease
- Author
-
Weihrauch, M., Re, D., Bischoff, S., Dietlein, M., Scheidhauer, K., Krug, B., Textoris, F., Ansén, S., Franklin, J., Bohlen, H., Wolf, J., Schicha, H., Diehl, V., and Tesch, H.
- Published
- 2002
- Full Text
- View/download PDF
3. Cancer genome diagnostics as prerequisite for an individualized treatment of lung cancer patients: V884
- Author
-
Ansén, S., Querings, S., Saal, B., Gabler, F., Seidel, D., Engel-Riedel, W., Stoelben, E., Markert, E., Zander, T., Wolf, J., and Thomas, R.
- Published
- 2009
4. Blood-based gene expression signatures for early detection of lung cancer: V607
- Author
-
Zander, T., Gaarz, A., Staratschek-Jox, A., Classeb, S., Henseler, M., Debey-Pascher, S., Eggle, D., Ansén, S., Massoudi, N., Stecki, M., Kraut, M., Frommolt, P., Hahn, M., Beyer, M., Thomas, R., Popov, A., Mauch, C., Delank, S., Stoelben, E., Engel-Riedel, W., Wichmann, E., Nagel, G., Linseise, J., Chang-Claude, J., Boffetta, P., Schultze, J. L., and Wolf, J.
- Published
- 2009
5. Chemical genomics of RAD001 in Non-Small Cell Lung Cancer: V606
- Author
-
Seidel, D., Ansén, S., Thomas, R., and Wolf, J.
- Published
- 2009
6. Myocardial Calcification following Septic Shock
- Author
-
Schellhammer, F., Ansén, S., Arnold, G., Brochhagen, H. G., and Lackner, K.
- Published
- 2002
- Full Text
- View/download PDF
7. Deutsche Hodgkin-Lymphom-Studiengruppe (DHSG): 20 Jahre Diagnostik, Therapie und Nachsorge von Patienten mit Morbus Hodgkin
- Author
-
Ansén, S., Engert, A., Wolf, J., Sieber, M., Paulus, U., and Diehl, V.
- Published
- 2001
8. Whole-body positron emission tomography using 18F-fluorodeoxyglucose for initial staging of patients with Hodgkin's disease
- Author
-
Weihrauch, M., primary, Re, D., additional, Bischoff, S., additional, Dietlein, M., additional, Scheidhauer, K., additional, Krug, B., additional, Textoris, F., additional, Ansén, S., additional, Franklin, J., additional, Bohlen, H., additional, Wolf, J., additional, Schicha, H., additional, Diehl, V., additional, and Tesch, H., additional
- Published
- 2001
- Full Text
- View/download PDF
9. Myocardial Calcification following Septic Shock.
- Author
-
Ansén, S., Arnold, G., Brochhagen, H.G., Lackner, K., and Schellhammer, F.
- Subjects
- *
LETTERS to the editor , *CARDIAC calcification - Abstract
Presents a letter to the editor in response to the article "Myocardial Calcification following Septic Shock" by F. Schellhammer, S. Ansén, G. Arnold, H.G. Brochhagen, and K. Lackner in the journal of Cardiology.
- Published
- 2002
- Full Text
- View/download PDF
10. Integrative genome analyses identify key somatic driver mutations of small-cell lung cancer
- Author
-
Jo Vandesompele, Peter Nürnberg, Shantanu Banerji, Lukas C. Heukamp, Stefanie Heynck, Matthias Fischer, Daniel Rauh, Sylvie Lantuejoul, Ingelore Baessmann, Holger Moch, Matthew Meyerson, Reinhard Büttner, Kwon-Sik Park, Ines Wilkening, Steinar Solberg, Stefan A. Haas, Egber Smit, Dennis Plenker, Zoe Wainer, Prudence A. Russell, Ilona Dahmen, William Pao, Erik Thunnissen, C. Ligorio, Bram De Wilde, Paul K. Brindle, Diana Böhm, Vito Michele Fazio, Vincenzo Di Cerbo, Benjamin Solomon, Stefania Damiani, Walburga Engel-Riedel, Erich Stoelben, Corinna Ludwig, Hannie Sietsma, Daniëlle A M Heideman, Jürgen Wolf, Thomas Muley, Elisabeth Brambilla, Ruping Sun, Wim Timens, Jay Shendure, Laura Pasqualucci, Kristian Cibulskis, Julien Sage, Gavin M. Wright, Mirjam Koker, Pierre Validire, Danila Seidel, Johannes M. Heuckmann, Harry J.M. Groen, Christian Becker, Philippe Lorimier, Peter J.F. Snijders, Sven Perner, Michael Brockmann, Xin Lu, Franziska Gabler, Scott L. Carter, Marius Lund-Iversen, Lucia Anna Muscarella, Jörg Sänger, Benjamin Besse, Hans Ulrich Schildhaus, Frauke Leenders, John K. Field, Odd Terje Brustugun, Christian Brambilla, Philipp A. Schnabel, Sascha Ansén, Christian Grütter, Michael Hallek, Gad Getz, Yuan Chen, Roopika Menon, Roman K. Thomas, Joachim H. Clement, Janine Altmüller, Martin L. Sos, Hans Hoffmann, Peter M. Schneider, Julie George, Christian Müller, Iver Petersen, Federico Cappuzzo, Lawryn H. Kasper, Robert Schneider, Martin Peifer, Lynnette Fernandez-Cuesta, Jean-Charles Soria, Alex Soltermann, Thomas Zander, Walter Weder, Pathology, Pulmonary medicine, CCA - Oncogenesis, Damage and Repair in Cancer Development and Cancer Treatment (DARE), Guided Treatment in Optimal Selected Cancer Patients (GUTS), Groningen Research Institute for Asthma and COPD (GRIAC), Peifer M, Fernández-Cuesta L, Sos ML, George J, Seidel D, Kasper LH, Plenker D, Leenders F, Sun R, Zander T, Menon R, Koker M, Dahmen I, Müller C, Di Cerbo V, Schildhaus HU, Altmüller J, Baessmann I, Becker C, de Wilde B, Vandesompele J, Böhm D, Ansén S, Gabler F, Wilkening I, Heynck S, Heuckmann JM, Lu X, Carter SL, Cibulskis K, Banerji S, Getz G, Park KS, Rauh D, Grütter C, Fischer M, Pasqualucci L, Wright G, Wainer Z, Russell P, Petersen I, Chen Y, Stoelben E, Ludwig C, Schnabel P, Hoffmann H, Muley T, Brockmann M, Engel-Riedel W, Muscarella LA, Fazio VM, Groen H, Timens W, Sietsma H, Thunnissen E, Smit E, Heideman DA, Snijders PJ, Cappuzzo F, Ligorio C, Damiani S, Field J, Solberg S, Brustugun OT, Lund-Iversen M, Sänger J, Clement JH, Soltermann A, Moch H, Weder W, Solomon B, Soria JC, Validire P, Besse B, Brambilla E, Brambilla C, Lantuejoul S, Lorimier P, Schneider PM, Hallek M, Pao W, Meyerson M, Sage J, Shendure J, Schneider R, Büttner R, Wolf J, Nürnberg P, Perner S, Heukamp LC, Brindle PK, Haas S, and Thomas RK.
- Subjects
Mutation rate ,EPH-RECEPTOR ,Genome ,Article ,lung ,03 medical and health sciences ,0302 clinical medicine ,E-CADHERIN ,Genetics ,PTEN ,EP300 ,small cell carcinoma ,neoplasms ,Exome sequencing ,ACUTE LYMPHOBLASTIC-LEUKEMIA ,030304 developmental biology ,P53 REGULATION ,0303 health sciences ,biology ,EGFR MUTATIONS ,MOUSE MODEL ,GENE ,humanities ,PROSTATE-CANCER ,respiratory tract diseases ,3. Good health ,FREQUENT MUTATION ,Gene expression profiling ,Histone ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Human genome ,NEUROENDOCRINE TUMORS - Abstract
Small-cell lung cancer (SCLC) is an aggressive lung tumor subtype with poor prognosis(1-3). We sequenced 29 SCLC exomes, 2 genomes and 15 transcriptomes and found an extremely high mutation rate of 7.4 +/- 1 protein-changing mutations per million base pairs. Therefore, we conducted integrated analyses of the various data sets to identify pathogenetically relevant mutated genes. In all cases, we found evidence for inactivation of TP53 and RB1 and identified recurrent mutations in the CREBBP, EP300 and MLL genes that encode histone modifiers. Furthermore, we observed mutations in PTEN, SLIT2 and EPHA7, as well as focal amplifications of the FGFR1 tyrosine kinase gene. Finally, we detected many of the alterations found in humans in SCLC tumors from Tp53 and Rb1 double knockout mice(4). Our study implicates histone modification as a major feature of SCLC, reveals potentially therapeutically tractable genomic alterations and provides a generalizable framework for the identification of biologically relevant genes in the context of high mutational background.
- Published
- 2012
11. Integrative genomic profiling of large-cell neuroendocrine carcinomas reveals distinct subtypes of high-grade neuroendocrine lung tumors.
- Author
-
George J, Walter V, Peifer M, Alexandrov LB, Seidel D, Leenders F, Maas L, Müller C, Dahmen I, Delhomme TM, Ardin M, Leblay N, Byrnes G, Sun R, De Reynies A, McLeer-Florin A, Bosco G, Malchers F, Menon R, Altmüller J, Becker C, Nürnberg P, Achter V, Lang U, Schneider PM, Bogus M, Soloway MG, Wilkerson MD, Cun Y, McKay JD, Moro-Sibilot D, Brambilla CG, Lantuejoul S, Lemaitre N, Soltermann A, Weder W, Tischler V, Brustugun OT, Lund-Iversen M, Helland Å, Solberg S, Ansén S, Wright G, Solomon B, Roz L, Pastorino U, Petersen I, Clement JH, Sänger J, Wolf J, Vingron M, Zander T, Perner S, Travis WD, Haas SA, Olivier M, Foll M, Büttner R, Hayes DN, Brambilla E, Fernandez-Cuesta L, and Thomas RK
- Subjects
- DNA Mutational Analysis, Genomics methods, High-Throughput Nucleotide Sequencing, Humans, Immunohistochemistry, In Situ Hybridization, Fluorescence, In Vitro Techniques, Lung Neoplasms genetics, Carcinoma, Neuroendocrine genetics, Carcinoma, Non-Small-Cell Lung genetics, Neuroendocrine Tumors genetics, Small Cell Lung Carcinoma genetics
- Abstract
Pulmonary large-cell neuroendocrine carcinomas (LCNECs) have similarities with other lung cancers, but their precise relationship has remained unclear. Here we perform a comprehensive genomic (n = 60) and transcriptomic (n = 69) analysis of 75 LCNECs and identify two molecular subgroups: "type I LCNECs" with bi-allelic TP53 and STK11/KEAP1 alterations (37%), and "type II LCNECs" enriched for bi-allelic inactivation of TP53 and RB1 (42%). Despite sharing genomic alterations with adenocarcinomas and squamous cell carcinomas, no transcriptional relationship was found; instead LCNECs form distinct transcriptional subgroups with closest similarity to SCLC. While type I LCNECs and SCLCs exhibit a neuroendocrine profile with ASCL1
high /DLL3high /NOTCHlow , type II LCNECs bear TP53 and RB1 alterations and differ from most SCLC tumors with reduced neuroendocrine markers, a pattern of ASCL1low /DLL3low /NOTCHhigh , and an upregulation of immune-related pathways. In conclusion, LCNECs comprise two molecularly defined subgroups, and distinguishing them from SCLC may allow stratified targeted treatment of high-grade neuroendocrine lung tumors.- Published
- 2018
- Full Text
- View/download PDF
12. Systematic Kinase Inhibitor Profiling Identifies CDK9 as a Synthetic Lethal Target in NUT Midline Carcinoma.
- Author
-
Brägelmann J, Dammert MA, Dietlein F, Heuckmann JM, Choidas A, Böhm S, Richters A, Basu D, Tischler V, Lorenz C, Habenberger P, Fang Z, Ortiz-Cuaran S, Leenders F, Eickhoff J, Koch U, Getlik M, Termathe M, Sallouh M, Greff Z, Varga Z, Balke-Want H, French CA, Peifer M, Reinhardt HC, Örfi L, Kéri G, Ansén S, Heukamp LC, Büttner R, Rauh D, Klebl BM, Thomas RK, and Sos ML
- Subjects
- Cell Cycle Proteins, Cell Line, Tumor, Cyclin T metabolism, Cyclin-Dependent Kinase 9 antagonists & inhibitors, Cyclin-Dependent Kinase 9 metabolism, HEK293 Cells, High-Throughput Screening Assays, Humans, Neoplasms genetics, Nuclear Proteins antagonists & inhibitors, Nuclear Proteins metabolism, Protein Kinase Inhibitors chemistry, RNA Polymerase II metabolism, Transcription Elongation, Genetic drug effects, Transcription Factors antagonists & inhibitors, Transcription Factors metabolism, Transcription, Genetic drug effects, Molecular Targeted Therapy, Neoplasms enzymology, Neoplasms pathology, Protein Kinase Inhibitors pharmacology
- Abstract
Kinase inhibitors represent the backbone of targeted cancer therapy, yet only a limited number of oncogenic drivers are directly druggable. By interrogating the activity of 1,505 kinase inhibitors, we found that BRD4-NUT-rearranged NUT midline carcinoma (NMC) cells are specifically killed by CDK9 inhibition (CDK9i) and depend on CDK9 and Cyclin-T1 expression. We show that CDK9i leads to robust induction of apoptosis and of markers of DNA damage response in NMC cells. While both CDK9i and bromodomain inhibition over time result in reduced Myc protein expression, only bromodomain inhibition induces cell differentiation and a p21-induced cell-cycle arrest in these cells. Finally, RNA-seq and ChIP-based analyses reveal a BRD4-NUT-specific CDK9i-induced perturbation of transcriptional elongation. Thus, our data provide a mechanistic basis for the genotype-dependent vulnerability of NMC cells to CDK9i that may be of relevance for the development of targeted therapies for NMC patients., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
13. Mechanisms of Primary Drug Resistance in FGFR1 -Amplified Lung Cancer.
- Author
-
Malchers F, Ercanoglu M, Schütte D, Castiglione R, Tischler V, Michels S, Dahmen I, Brägelmann J, Menon R, Heuckmann JM, George J, Ansén S, Sos ML, Soltermann A, Peifer M, Wolf J, Büttner R, and Thomas RK
- Subjects
- Animals, Antineoplastic Agents pharmacology, Biomarkers, Tumor, Cell Line, Tumor, Disease Models, Animal, GTP Phosphohydrolases genetics, Humans, Lung Neoplasms diagnosis, Lung Neoplasms drug therapy, Membrane Proteins genetics, Mice, Protein Binding, Protein Kinase Inhibitors pharmacology, Proto-Oncogene Proteins c-met, Receptor, Fibroblast Growth Factor, Type 1 antagonists & inhibitors, Tomography, X-Ray Computed, Translocation, Genetic, Xenograft Model Antitumor Assays, Drug Resistance, Neoplasm genetics, Gene Amplification, Lung Neoplasms genetics, Receptor, Fibroblast Growth Factor, Type 1 genetics
- Abstract
Purpose: The 8p12-p11 locus is frequently amplified in squamous cell lung cancer (SQLC); the receptor tyrosine kinase fibroblast growth factor receptor 1 (FGFR1) being one of the most prominent targets of this amplification. Thus, small molecules inhibiting FGFRs have been employed to treat FGFR1 -amplified SQLC. However, only about 11% of such FGFR1 -amplified tumors respond to single-agent FGFR inhibition and several tumors exhibited insufficient tumor shrinkage, compatible with the existence of drug-resistant tumor cells. Experimental Design: To investigate possible mechanisms of resistance to FGFR inhibition, we studied the lung cancer cell lines DMS114 and H1581. Both cell lines are highly sensitive to three different FGFR inhibitors, but exhibit sustained residual cellular viability under treatment, indicating a subpopulation of existing drug-resistant cells. We isolated these subpopulations by treating the cells with constant high doses of FGFR inhibitors. Results: The FGFR inhibitor-resistant cells were cross-resistant and characterized by sustained MAPK pathway activation. In drug-resistant H1581 cells, we identified NRAS amplification and DUSP6 deletion, leading to MAPK pathway reactivation. Furthermore, we detected subclonal NRAS amplifications in 3 of 20 (15%) primary human FGFR1 -amplified SQLC specimens. In contrast, drug-resistant DMS114 cells exhibited transcriptional upregulation of MET that drove MAPK pathway reactivation. As a consequence, we demonstrate that rational combination therapies resensitize resistant cells to treatment with FGFR inhibitors. Conclusions: We provide evidence for the existence of diverse mechanisms of primary drug resistance in FGFR1 -amplified lung cancer and provide a rational strategy to improve FGFR inhibitor therapies by combination treatment. Clin Cancer Res; 23(18); 5527-36. ©2017 AACR ., (©2017 American Association for Cancer Research.)
- Published
- 2017
- Full Text
- View/download PDF
14. Heterogeneous Mechanisms of Primary and Acquired Resistance to Third-Generation EGFR Inhibitors.
- Author
-
Ortiz-Cuaran S, Scheffler M, Plenker D, Dahmen L, Scheel AH, Fernandez-Cuesta L, Meder L, Lovly CM, Persigehl T, Merkelbach-Bruse S, Bos M, Michels S, Fischer R, Albus K, König K, Schildhaus HU, Fassunke J, Ihle MA, Pasternack H, Heydt C, Becker C, Altmüller J, Ji H, Müller C, Florin A, Heuckmann JM, Nuernberg P, Ansén S, Heukamp LC, Berg J, Pao W, Peifer M, Buettner R, Wolf J, Thomas RK, and Sos ML
- Subjects
- Acrylamides therapeutic use, Adenocarcinoma of Lung, Aged, Aniline Compounds therapeutic use, ErbB Receptors antagonists & inhibitors, Female, Humans, Male, Middle Aged, Pyrimidines therapeutic use, Adenocarcinoma drug therapy, Antineoplastic Agents therapeutic use, Drug Resistance, Neoplasm genetics, Lung Neoplasms drug therapy
- Abstract
Purpose: To identify novel mechanisms of resistance to third-generation EGFR inhibitors in patients with lung adenocarcinoma that progressed under therapy with either AZD9291 or rociletinib (CO-1686)., Experimental Design: We analyzed tumor biopsies from seven patients obtained before, during, and/or after treatment with AZD9291 or rociletinib (CO-1686). Targeted sequencing and FISH analyses were performed, and the relevance of candidate genes was functionally assessed in in vitro models., Results: We found recurrent amplification of either MET or ERBB2 in tumors that were resistant or developed resistance to third-generation EGFR inhibitors and show that ERBB2 and MET activation can confer resistance to these compounds. Furthermore, we identified a KRAS
G12S mutation in a patient with acquired resistance to AZD9291 as a potential driver of acquired resistance. Finally, we show that dual inhibition of EGFR/MEK might be a viable strategy to overcome resistance in EGFR-mutant cells expressing mutant KRAS CONCLUSIONS: Our data suggest that heterogeneous mechanisms of resistance can drive primary and acquired resistance to third-generation EGFR inhibitors and provide a rationale for potential combination strategies. Clin Cancer Res; 22(19); 4837-47. ©2016 AACR., (©2016 American Association for Cancer Research.)- Published
- 2016
- Full Text
- View/download PDF
15. PD-L1 expression in non-small cell lung cancer: Correlations with genetic alterations.
- Author
-
Scheel AH, Ansén S, Schultheis AM, Scheffler M, Fischer RN, Michels S, Hellmich M, George J, Zander T, Brockmann M, Stoelben E, Groen H, Timens W, Perner S, von Bergwelt-Baildon M, Büttner R, and Wolf J
- Abstract
Inhibition of the PD-1/PD-L1 pathway may induce anticancer immune responses in non-small cell lung cancer (NSCLC). Two PD-L1 immunohistochemistry (IHC) assays have been approved as companion diagnostic tests for therapeutic anti-PD-1 antibodies. However, many aspects of PD-L1 prevalence and association with genetically defined subtypes have not been addressed systematically. Here, we analyzed PD-L1 expression in 436 genetically annotated NSCLC specimens enriched for early stages using PD-L1 antibody 5H1. Expression of PD-L1 was detected in the tumor cells (TC) (34% of cases) and in associated immune cells (IC) (49%) across all stages of NSCLC, either alone or in combination. PD-L1 IHC-positive TC, but not IC showed significantly higher PD-L1 RNA expression levels. Expression in TC was associated with TP53, KRAS and STK11 mutational status in adenocarcinomas (AD) and with NFE2L2 mutations in squamous cell carcinomas (SQ). No correlations with histological subtype, clinical characteristics and overall survival were found. The presence of PD-L1-positive IC was significantly associated with patients' smoking status in AD. The findings are in agreement with the emerging concept that tumors with high mutational burden are more likely to benefit from immunotherapy, since TP53 and KRAS mutations are linked to smoking, increased numbers of somatic mutations and expression of neoantigens. Current clinical studies focus on stage IIIB and IV NSCLC; however, PD-L1 expression occurs in earlier stages and might be a predictive biomarker in clinical trials testing (neo-) adjuvant strategies.
- Published
- 2016
- Full Text
- View/download PDF
16. Comprehensive genomic profiles of small cell lung cancer.
- Author
-
George J, Lim JS, Jang SJ, Cun Y, Ozretić L, Kong G, Leenders F, Lu X, Fernández-Cuesta L, Bosco G, Müller C, Dahmen I, Jahchan NS, Park KS, Yang D, Karnezis AN, Vaka D, Torres A, Wang MS, Korbel JO, Menon R, Chun SM, Kim D, Wilkerson M, Hayes N, Engelmann D, Pützer B, Bos M, Michels S, Vlasic I, Seidel D, Pinther B, Schaub P, Becker C, Altmüller J, Yokota J, Kohno T, Iwakawa R, Tsuta K, Noguchi M, Muley T, Hoffmann H, Schnabel PA, Petersen I, Chen Y, Soltermann A, Tischler V, Choi CM, Kim YH, Massion PP, Zou Y, Jovanovic D, Kontic M, Wright GM, Russell PA, Solomon B, Koch I, Lindner M, Muscarella LA, la Torre A, Field JK, Jakopovic M, Knezevic J, Castaños-Vélez E, Roz L, Pastorino U, Brustugun OT, Lund-Iversen M, Thunnissen E, Köhler J, Schuler M, Botling J, Sandelin M, Sanchez-Cespedes M, Salvesen HB, Achter V, Lang U, Bogus M, Schneider PM, Zander T, Ansén S, Hallek M, Wolf J, Vingron M, Yatabe Y, Travis WD, Nürnberg P, Reinhardt C, Perner S, Heukamp L, Büttner R, Haas SA, Brambilla E, Peifer M, Sage J, and Thomas RK
- Subjects
- Alleles, Animals, Cell Line, Tumor, Chromosome Breakpoints, Cyclin D1 genetics, DNA-Binding Proteins genetics, Disease Models, Animal, Female, Gene Expression Profiling, Humans, Lung Neoplasms metabolism, Lung Neoplasms pathology, Male, Mice, Neurosecretory Systems metabolism, Neurosecretory Systems pathology, Nuclear Proteins genetics, Receptors, Notch genetics, Receptors, Notch metabolism, Retinoblastoma Protein genetics, Signal Transduction genetics, Small Cell Lung Carcinoma metabolism, Small Cell Lung Carcinoma pathology, Tumor Protein p73, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Proteins genetics, Genome, Human genetics, Genomics, Lung Neoplasms genetics, Mutation genetics, Small Cell Lung Carcinoma genetics
- Abstract
We have sequenced the genomes of 110 small cell lung cancers (SCLC), one of the deadliest human cancers. In nearly all the tumours analysed we found bi-allelic inactivation of TP53 and RB1, sometimes by complex genomic rearrangements. Two tumours with wild-type RB1 had evidence of chromothripsis leading to overexpression of cyclin D1 (encoded by the CCND1 gene), revealing an alternative mechanism of Rb1 deregulation. Thus, loss of the tumour suppressors TP53 and RB1 is obligatory in SCLC. We discovered somatic genomic rearrangements of TP73 that create an oncogenic version of this gene, TP73Δex2/3. In rare cases, SCLC tumours exhibited kinase gene mutations, providing a possible therapeutic opportunity for individual patients. Finally, we observed inactivating mutations in NOTCH family genes in 25% of human SCLC. Accordingly, activation of Notch signalling in a pre-clinical SCLC mouse model strikingly reduced the number of tumours and extended the survival of the mutant mice. Furthermore, neuroendocrine gene expression was abrogated by Notch activity in SCLC cells. This first comprehensive study of somatic genome alterations in SCLC uncovers several key biological processes and identifies candidate therapeutic targets in this highly lethal form of cancer.
- Published
- 2015
- Full Text
- View/download PDF
17. Identification of novel fusion genes in lung cancer using breakpoint assembly of transcriptome sequencing data.
- Author
-
Fernandez-Cuesta L, Sun R, Menon R, George J, Lorenz S, Meza-Zepeda LA, Peifer M, Plenker D, Heuckmann JM, Leenders F, Zander T, Dahmen I, Koker M, Schöttle J, Ullrich RT, Altmüller J, Becker C, Nürnberg P, Seidel H, Böhm D, Göke F, Ansén S, Russell PA, Wright GM, Wainer Z, Solomon B, Petersen I, Clement JH, Sänger J, Brustugun OT, Helland Å, Solberg S, Lund-Iversen M, Buettner R, Wolf J, Brambilla E, Vingron M, Perner S, Haas SA, and Thomas RK
- Subjects
- Base Sequence, Cell Line, Tumor, Cluster Analysis, Gene Silencing, Genomics, Humans, In Situ Hybridization, Fluorescence, Lung Neoplasms genetics, Oncogene Proteins, Fusion genetics, Tumor Suppressor Proteins genetics, Chromosome Breakpoints, Computational Biology methods, High-Throughput Nucleotide Sequencing, Oncogene Fusion, Transcriptome, Translocation, Genetic
- Abstract
Genomic translocation events frequently underlie cancer development through generation of gene fusions with oncogenic properties. Identification of such fusion transcripts by transcriptome sequencing might help to discover new potential therapeutic targets. We developed TRUP (Tumor-specimen suited RNA-seq Unified Pipeline) (https://github.com/ruping/TRUP), a computational approach that combines split-read and read-pair analysis with de novo assembly for the identification of chimeric transcripts in cancer specimens. We apply TRUP to RNA-seq data of different tumor types, and find it to be more sensitive than alternative tools in detecting chimeric transcripts, such as secondary rearrangements in EML4-ALK-positive lung tumors, or recurrent inactivating rearrangements affecting RASSF8.
- Published
- 2015
- Full Text
- View/download PDF
18. Rationale for co-targeting IGF-1R and ALK in ALK fusion-positive lung cancer.
- Author
-
Lovly CM, McDonald NT, Chen H, Ortiz-Cuaran S, Heukamp LC, Yan Y, Florin A, Ozretić L, Lim D, Wang L, Chen Z, Chen X, Lu P, Paik PK, Shen R, Jin H, Buettner R, Ansén S, Perner S, Brockmann M, Bos M, Wolf J, Gardizi M, Wright GM, Solomon B, Russell PA, Rogers TM, Suehara Y, Red-Brewer M, Tieu R, de Stanchina E, Wang Q, Zhao Z, Johnson DH, Horn L, Wong KK, Thomas RK, Ladanyi M, and Pao W
- Subjects
- Anaplastic Lymphoma Kinase, Crizotinib, Female, Gene Knockdown Techniques, Humans, Insulin Receptor Substrate Proteins genetics, Lung Neoplasms enzymology, Lung Neoplasms metabolism, Middle Aged, Pyrazoles pharmacology, Pyridines pharmacology, Receptor, IGF Type 1 metabolism, Up-Regulation, Lung Neoplasms drug therapy, Pyrazoles therapeutic use, Pyridines therapeutic use, Receptor Protein-Tyrosine Kinases drug effects, Receptor, IGF Type 1 drug effects
- Abstract
Crizotinib, a selective tyrosine kinase inhibitor (TKI), shows marked activity in patients whose lung cancers harbor fusions in the gene encoding anaplastic lymphoma receptor tyrosine kinase (ALK), but its efficacy is limited by variable primary responses and acquired resistance. In work arising from the clinical observation of a patient with ALK fusion-positive lung cancer who had an exceptional response to an insulin-like growth factor 1 receptor (IGF-1R)-specific antibody, we define a therapeutic synergism between ALK and IGF-1R inhibitors. Similar to IGF-1R, ALK fusion proteins bind to the adaptor insulin receptor substrate 1 (IRS-1), and IRS-1 knockdown enhances the antitumor effects of ALK inhibitors. In models of ALK TKI resistance, the IGF-1R pathway is activated, and combined ALK and IGF-1R inhibition improves therapeutic efficacy. Consistent with this finding, the levels of IGF-1R and IRS-1 are increased in biopsy samples from patients progressing on crizotinib monotherapy. Collectively these data support a role for the IGF-1R-IRS-1 pathway in both ALK TKI-sensitive and ALK TKI-resistant states and provide a biological rationale for further clinical development of dual ALK and IGF-1R inhibitors.
- Published
- 2014
- Full Text
- View/download PDF
19. Rapid remineralization of multiple disseminated bone lesions after high-dose cytarabine in a patient with isolated myeloid sarcoma.
- Author
-
Brähler S, Thielen I, Schwabe H, Engels M, Kreuzer KA, Wolf J, and Ansén S
- Subjects
- Antimetabolites, Antineoplastic adverse effects, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Biopsy, Bone Diseases diagnosis, Bone Marrow pathology, Calcinosis diagnosis, Cytarabine adverse effects, Humans, Immunophenotyping, Male, Middle Aged, Sarcoma, Myeloid diagnosis, Tomography, X-Ray Computed, Treatment Outcome, Antimetabolites, Antineoplastic administration & dosage, Bone Diseases etiology, Calcinosis etiology, Cytarabine administration & dosage, Sarcoma, Myeloid complications, Sarcoma, Myeloid drug therapy
- Abstract
Isolated myeloid sarcoma is a rare presentation of acute myeloid leukemia. There are limited data available concerning the prognostic relevance and the right treatment strategy for this clinical scenario. Here, we report a case of acute myeloid leukemia with extensive lesions and fractures in multiple bones in a 64-yr-old male patient. Remarkably, treatment with a high-dose cytarabine regimen led to rapid remineralization of all bone lesions and recovery of the patient's mobility within a few weeks. Thereby, surgical treatment and radiotherapy could be avoided, supporting the role of intensive induction and standard consolidation chemotherapy as first-line treatment for myeloid sarcoma., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2014
- Full Text
- View/download PDF
20. CD74-NRG1 fusions in lung adenocarcinoma.
- Author
-
Fernandez-Cuesta L, Plenker D, Osada H, Sun R, Menon R, Leenders F, Ortiz-Cuaran S, Peifer M, Bos M, Daßler J, Malchers F, Schöttle J, Vogel W, Dahmen I, Koker M, Ullrich RT, Wright GM, Russell PA, Wainer Z, Solomon B, Brambilla E, Nagy-Mignotte H, Moro-Sibilot D, Brambilla CG, Lantuejoul S, Altmüller J, Becker C, Nürnberg P, Heuckmann JM, Stoelben E, Petersen I, Clement JH, Sänger J, Muscarella LA, la Torre A, Fazio VM, Lahortiga I, Perera T, Ogata S, Parade M, Brehmer D, Vingron M, Heukamp LC, Buettner R, Zander T, Wolf J, Perner S, Ansén S, Haas SA, Yatabe Y, and Thomas RK
- Subjects
- Adenocarcinoma pathology, Adenocarcinoma of Lung, Adenocarcinoma, Mucinous pathology, Adult, Aged, Aged, 80 and over, Animals, Base Sequence, Cell Line, Tumor, Female, Gene Expression Profiling, Humans, Lung Neoplasms pathology, Male, Mice, Middle Aged, Molecular Sequence Data, NIH 3T3 Cells, Oncogene Proteins, Fusion metabolism, Sequence Analysis, DNA, Signal Transduction genetics, Adenocarcinoma genetics, Adenocarcinoma, Mucinous genetics, Antigens, Differentiation, B-Lymphocyte genetics, Histocompatibility Antigens Class II genetics, Lung Neoplasms genetics, Neuregulin-1 genetics, Oncogene Proteins, Fusion genetics
- Abstract
Unlabelled: We discovered a novel somatic gene fusion, CD74-NRG1, by transcriptome sequencing of 25 lung adenocarcinomas of never smokers. By screening 102 lung adenocarcinomas negative for known oncogenic alterations, we found four additional fusion-positive tumors, all of which were of the invasive mucinous subtype. Mechanistically, CD74-NRG1 leads to extracellular expression of the EGF-like domain of NRG1 III-β3, thereby providing the ligand for ERBB2-ERBB3 receptor complexes. Accordingly, ERBB2 and ERBB3 expression was high in the index case, and expression of phospho-ERBB3 was specifically found in tumors bearing the fusion (P < 0.0001). Ectopic expression of CD74-NRG1 in lung cancer cell lines expressing ERBB2 and ERBB3 activated ERBB3 and the PI3K-AKT pathway, and led to increased colony formation in soft agar. Thus, CD74-NRG1 gene fusions are activating genomic alterations in invasive mucinous adenocarcinomas and may offer a therapeutic opportunity for a lung tumor subtype with, so far, no effective treatment., Significance: CD74–NRG1 fusions may represent a therapeutic opportunity for invasive mucinous lung adenocarcinomas, a tumor with no effective treatment that frequently presents with multifocal unresectable disease.
- Published
- 2014
- Full Text
- View/download PDF
21. Frequent mutations in chromatin-remodelling genes in pulmonary carcinoids.
- Author
-
Fernandez-Cuesta L, Peifer M, Lu X, Sun R, Ozretić L, Seidal D, Zander T, Leenders F, George J, Müller C, Dahmen I, Pinther B, Bosco G, Konrad K, Altmüller J, Nürnberg P, Achter V, Lang U, Schneider PM, Bogus M, Soltermann A, Brustugun OT, Helland Å, Solberg S, Lund-Iversen M, Ansén S, Stoelben E, Wright GM, Russell P, Wainer Z, Solomon B, Field JK, Hyde R, Davies MP, Heukamp LC, Petersen I, Perner S, Lovly C, Cappuzzo F, Travis WD, Wolf J, Vingron M, Brambilla E, Haas SA, Buettner R, and Thomas RK
- Subjects
- Adolescent, Adult, Aged, Base Sequence, Carcinoid Tumor pathology, Chromosome Mapping, DNA Copy Number Variations, Exome genetics, Female, Gene Dosage, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Genome, Human genetics, Humans, Lung Neoplasms pathology, Male, Middle Aged, Molecular Sequence Data, Polymorphism, Single Nucleotide, Sequence Analysis, DNA, Young Adult, Carcinoid Tumor genetics, Chromatin Assembly and Disassembly genetics, Lung Neoplasms genetics, Mutation
- Abstract
Pulmonary carcinoids are rare neuroendocrine tumours of the lung. The molecular alterations underlying the pathogenesis of these tumours have not been systematically studied so far. Here we perform gene copy number analysis (n=54), genome/exome (n=44) and transcriptome (n=69) sequencing of pulmonary carcinoids and observe frequent mutations in chromatin-remodelling genes. Covalent histone modifiers and subunits of the SWI/SNF complex are mutated in 40 and 22.2% of the cases, respectively, with MEN1, PSIP1 and ARID1A being recurrently affected. In contrast to small-cell lung cancer and large-cell neuroendocrine lung tumours, TP53 and RB1 mutations are rare events, suggesting that pulmonary carcinoids are not early progenitor lesions of the highly aggressive lung neuroendocrine tumours but arise through independent cellular mechanisms. These data also suggest that inactivation of chromatin-remodelling genes is sufficient to drive transformation in pulmonary carcinoids.
- Published
- 2014
- Full Text
- View/download PDF
22. Definition of a fluorescence in-situ hybridization score identifies high- and low-level FGFR1 amplification types in squamous cell lung cancer.
- Author
-
Schildhaus HU, Heukamp LC, Merkelbach-Bruse S, Riesner K, Schmitz K, Binot E, Paggen E, Albus K, Schulte W, Ko YD, Schlesinger A, Ansén S, Engel-Riedel W, Brockmann M, Serke M, Gerigk U, Huss S, Göke F, Perner S, Hekmat K, Frank KF, Reiser M, Schnell R, Bos M, Mattonet C, Sos M, Stoelben E, Wolf J, Zander T, and Buettner R
- Subjects
- Adenocarcinoma pathology, Adenocarcinoma of Lung, Carcinoma, Squamous Cell pathology, Fixatives, Formaldehyde, Gene Dosage, Genetic Predisposition to Disease, Germany, Humans, Lung Neoplasms pathology, Paraffin Embedding, Phenotype, Predictive Value of Tests, Reproducibility of Results, Tissue Fixation, Adenocarcinoma genetics, Carcinoma, Squamous Cell genetics, Gene Amplification, In Situ Hybridization, Fluorescence, Lung Neoplasms genetics, Receptor, Fibroblast Growth Factor, Type 1 genetics
- Abstract
We recently reported fibroblast growth factor receptor-type 1 (FGFR1) amplification to be associated with therapeutically tractable FGFR1 dependency in squamous cell lung cancer. This makes FGFR1 a novel target for directed therapy in these tumors. To reproducibly identify patients for clinical studies, we developed a standardized reading and evaluation strategy for FGFR1 fluorescence in-situ hybridization (FISH) and propose evaluation criteria, describe different patterns of low- and high-level amplifications and report on the prevalence of FGFR1 amplifications in pulmonary carcinomas. A total of 420 lung cancer patients including 307 squamous carcinomas, 100 adenocarcinomas of the lung and 13 carcinomas of other types were analyzed for FGFR1 amplification using a dual color FISH. We found heterogeneous and different patterns of gene copy numbers. FGFR1 amplifications were observed in 20% of pulmonary squamous carcinomas but not in adenocarcinomas. High-level amplification (as defined by an FGFR1/centromer 8 (CEN8) ratio ≥2.0, or average number of FGFR1 signals per tumor cell nucleus ≥6, or the percentage of tumor cells containing ≥15 FGFR1 signals or large clusters ≥10%) was detected at a frequency of 16% and low-level amplification (as defined by ≥5 FGFR1 signals in ≥50% of tumor cells) at a frequency of 4%. We conclude that FGFR1 amplification is one of the most frequent therapeutically tractable genetic lesions in pulmonary carcinomas. Standardized reporting of FGFR1 amplification in squamous carcinomas of the lung will become increasingly important to correlate therapeutic responses with FGFR1 inhibitors in clinical studies. Thus, our reading and evaluation strategy might serve as a basis for identifying patients for ongoing and upcoming clinical trials.
- Published
- 2012
- Full Text
- View/download PDF
23. Integrative genome analyses identify key somatic driver mutations of small-cell lung cancer.
- Author
-
Peifer M, Fernández-Cuesta L, Sos ML, George J, Seidel D, Kasper LH, Plenker D, Leenders F, Sun R, Zander T, Menon R, Koker M, Dahmen I, Müller C, Di Cerbo V, Schildhaus HU, Altmüller J, Baessmann I, Becker C, de Wilde B, Vandesompele J, Böhm D, Ansén S, Gabler F, Wilkening I, Heynck S, Heuckmann JM, Lu X, Carter SL, Cibulskis K, Banerji S, Getz G, Park KS, Rauh D, Grütter C, Fischer M, Pasqualucci L, Wright G, Wainer Z, Russell P, Petersen I, Chen Y, Stoelben E, Ludwig C, Schnabel P, Hoffmann H, Muley T, Brockmann M, Engel-Riedel W, Muscarella LA, Fazio VM, Groen H, Timens W, Sietsma H, Thunnissen E, Smit E, Heideman DA, Snijders PJ, Cappuzzo F, Ligorio C, Damiani S, Field J, Solberg S, Brustugun OT, Lund-Iversen M, Sänger J, Clement JH, Soltermann A, Moch H, Weder W, Solomon B, Soria JC, Validire P, Besse B, Brambilla E, Brambilla C, Lantuejoul S, Lorimier P, Schneider PM, Hallek M, Pao W, Meyerson M, Sage J, Shendure J, Schneider R, Büttner R, Wolf J, Nürnberg P, Perner S, Heukamp LC, Brindle PK, Haas S, and Thomas RK
- Subjects
- Amino Acid Substitution, Animals, CREB-Binding Protein genetics, Cell Line, Tumor, DNA Copy Number Variations, DNA Mutational Analysis, E1A-Associated p300 Protein genetics, Gene Expression Profiling, Gene Regulatory Networks, Genome-Wide Association Study, Histone-Lysine N-Methyltransferase, Humans, Intercellular Signaling Peptides and Proteins genetics, Mice, Mice, Knockout, Models, Molecular, Mutation, Myeloid-Lymphoid Leukemia Protein genetics, Nerve Tissue Proteins genetics, Oligonucleotide Array Sequence Analysis, PTEN Phosphohydrolase genetics, Polymorphism, Single Nucleotide, Protein Processing, Post-Translational genetics, Retinoblastoma Protein genetics, Tumor Suppressor Protein p53 genetics, Genome, Human, Lung Neoplasms genetics, Small Cell Lung Carcinoma genetics
- Abstract
Small-cell lung cancer (SCLC) is an aggressive lung tumor subtype with poor prognosis. We sequenced 29 SCLC exomes, 2 genomes and 15 transcriptomes and found an extremely high mutation rate of 7.4±1 protein-changing mutations per million base pairs. Therefore, we conducted integrated analyses of the various data sets to identify pathogenetically relevant mutated genes. In all cases, we found evidence for inactivation of TP53 and RB1 and identified recurrent mutations in the CREBBP, EP300 and MLL genes that encode histone modifiers. Furthermore, we observed mutations in PTEN, SLIT2 and EPHA7, as well as focal amplifications of the FGFR1 tyrosine kinase gene. Finally, we detected many of the alterations found in humans in SCLC tumors from Tp53 and Rb1 double knockout mice. Our study implicates histone modification as a major feature of SCLC, reveals potentially therapeutically tractable genomic alterations and provides a generalizable framework for the identification of biologically relevant genes in the context of high mutational background.
- Published
- 2012
- Full Text
- View/download PDF
24. Mapping the hallmarks of lung adenocarcinoma with massively parallel sequencing.
- Author
-
Imielinski M, Berger AH, Hammerman PS, Hernandez B, Pugh TJ, Hodis E, Cho J, Suh J, Capelletti M, Sivachenko A, Sougnez C, Auclair D, Lawrence MS, Stojanov P, Cibulskis K, Choi K, de Waal L, Sharifnia T, Brooks A, Greulich H, Banerji S, Zander T, Seidel D, Leenders F, Ansén S, Ludwig C, Engel-Riedel W, Stoelben E, Wolf J, Goparju C, Thompson K, Winckler W, Kwiatkowski D, Johnson BE, Jänne PA, Miller VA, Pao W, Travis WD, Pass HI, Gabriel SB, Lander ES, Thomas RK, Garraway LA, Getz G, and Meyerson M
- Subjects
- Adenocarcinoma pathology, Adenocarcinoma of Lung, Adult, Aged, Aged, 80 and over, Carcinoma, Non-Small-Cell Lung pathology, Cohort Studies, Exome, Female, Genome-Wide Association Study, Humans, Lung Neoplasms pathology, Male, Middle Aged, Mutation, Mutation Rate, Adenocarcinoma genetics, Carcinoma, Non-Small-Cell Lung genetics, Genes, Neoplasm, High-Throughput Nucleotide Sequencing, Lung Neoplasms genetics
- Abstract
Lung adenocarcinoma, the most common subtype of non-small cell lung cancer, is responsible for more than 500,000 deaths per year worldwide. Here, we report exome and genome sequences of 183 lung adenocarcinoma tumor/normal DNA pairs. These analyses revealed a mean exonic somatic mutation rate of 12.0 events/megabase and identified the majority of genes previously reported as significantly mutated in lung adenocarcinoma. In addition, we identified statistically recurrent somatic mutations in the splicing factor gene U2AF1 and truncating mutations affecting RBM10 and ARID1A. Analysis of nucleotide context-specific mutation signatures grouped the sample set into distinct clusters that correlated with smoking history and alterations of reported lung adenocarcinoma genes. Whole-genome sequence analysis revealed frequent structural rearrangements, including in-frame exonic alterations within EGFR and SIK2 kinases. The candidate genes identified in this study are attractive targets for biological characterization and therapeutic targeting of lung adenocarcinoma., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
25. IL-21 can supplement suboptimal Lck-independent MAPK activation in a STAT-3-dependent manner in human CD8(+) T cells.
- Author
-
Imataki O, Ansén S, Tanaka M, Butler MO, Berezovskaya A, Milstein MI, Kuzushima K, Nadler LM, and Hirano N
- Subjects
- CD8 Antigens immunology, CD8-Positive T-Lymphocytes metabolism, Cells, Cultured, Humans, Interleukins immunology, Lymphocyte Activation, Major Histocompatibility Complex, Signal Transduction, T-Lymphocytes, Cytotoxic metabolism, CD8-Positive T-Lymphocytes immunology, Interleukins metabolism, Lymphocyte Specific Protein Tyrosine Kinase p56(lck) metabolism, Mitogen-Activated Protein Kinases metabolism, STAT3 Transcription Factor metabolism, T-Lymphocytes, Cytotoxic immunology
- Abstract
Although both MHC class II/CD8α double-knockout and CD8β null mice show a defect in the development of MHC class I-restricted CD8(+) T cells in the thymus, they possess low numbers of high-avidity peripheral CTL with limited clonality and are able to contain acute and chronic infections. These in vivo data suggest that the CD8 coreceptor is not absolutely necessary for the generation of Ag-specific CTL. Lack of CD8 association causes partial TCR signaling because of the absence of CD8/Lck recruitment to the proximity of the MHC/TCR complex, resulting in suboptimal MAPK activation. Therefore, there should exist a signaling mechanism that can supplement partial TCR activation caused by the lack of CD8 association. In this human study, we have shown that CD8-independent stimulation of Ag-specific CTL previously primed in the presence of CD8 coligation, either in vivo or in vitro, induced severely impaired in vitro proliferation. When naive CD8(+) T cells were primed in the absence of CD8 binding and subsequently restimulated in the presence of CD8 coligation, the proliferation of Ag-specific CTL was also severely hampered. However, when CD8-independent T cell priming and restimulation were supplemented with IL-21, Ag-specific CD8(+) CTL expanded in two of six individuals tested. We found that IL-21 rescued partial MAPK activation in a STAT3- but not STAT1-dependent manner. These results suggest that CD8 coligation is critical for the expansion of postthymic peripheral Ag-specific CTL in humans. However, STAT3-mediated IL-21 signaling can supplement partial TCR signaling caused by the lack of CD8 association.
- Published
- 2012
- Full Text
- View/download PDF
26. Ex vivo expansion of human CD8+ T cells using autologous CD4+ T cell help.
- Author
-
Butler MO, Imataki O, Yamashita Y, Tanaka M, Ansén S, Berezovskaya A, Metzler G, Milstein MI, Mooney MM, Murray AP, Mano H, Nadler LM, and Hirano N
- Subjects
- CD3 Complex immunology, CD3 Complex metabolism, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes metabolism, Cells, Cultured, Coculture Techniques, Cytokines immunology, Cytokines metabolism, Flow Cytometry, Humans, Interferon-gamma immunology, Interferon-gamma metabolism, Interleukin-2 immunology, Interleukin-2 metabolism, Interleukin-2 pharmacology, Interleukins immunology, Interleukins metabolism, Interleukins pharmacology, K562 Cells, Lymphocytes, Tumor-Infiltrating drug effects, Lymphocytes, Tumor-Infiltrating immunology, Lymphocytes, Tumor-Infiltrating metabolism, Muromonab-CD3 immunology, Muromonab-CD3 metabolism, Muromonab-CD3 pharmacology, Receptors, Interleukin-21 immunology, Receptors, Interleukin-21 metabolism, T-Lymphocytes immunology, T-Lymphocytes metabolism, Up-Regulation drug effects, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Cell Proliferation
- Abstract
Background: Using in vivo mouse models, the mechanisms of CD4+ T cell help have been intensively investigated. However, a mechanistic analysis of human CD4+ T cell help is largely lacking. Our goal was to elucidate the mechanisms of human CD4+ T cell help of CD8+ T cell proliferation using a novel in vitro model., Methods/principal Findings: We developed a genetically engineered novel human cell-based artificial APC, aAPC/mOKT3, which expresses a membranous form of the anti-CD3 monoclonal antibody OKT3 as well as other immune accessory molecules. Without requiring the addition of allogeneic feeder cells, aAPC/mOKT3 enabled the expansion of both peripheral and tumor-infiltrating T cells, regardless of HLA-restriction. Stimulation with aAPC/mOKT3 did not expand Foxp3+ regulatory T cells, and expanded tumor infiltrating lymphocytes predominantly secreted Th1-type cytokines, interferon-γ and IL-2. In this aAPC-based system, the presence of autologous CD4+ T cells was associated with significantly improved CD8+ T cell expansion in vitro. The CD4+ T cell derived cytokines IL-2 and IL-21 were necessary but not sufficient for this effect. However, CD4+ T cell help of CD8+ T cell proliferation was partially recapitulated by both adding IL-2/IL-21 and by upregulation of IL-21 receptor on CD8+ T cells., Conclusions: We have developed an in vitro model that advances our understanding of the immunobiology of human CD4+ T cell help of CD8+ T cells. Our data suggests that human CD4+ T cell help can be leveraged to expand CD8+ T cells in vitro.
- Published
- 2012
- Full Text
- View/download PDF
27. Induction of HLA-DP4-restricted anti-survivin Th1 and Th2 responses using an artificial antigen-presenting cell.
- Author
-
Tanaka M, Butler MO, Ansén S, Imataki O, Berezovskaya A, Nadler LM, and Hirano N
- Subjects
- Alleles, Amino Acid Sequence, Antigen-Presenting Cells metabolism, Antigens, CD immunology, Antigens, CD metabolism, B7-1 Antigen immunology, B7-1 Antigen metabolism, Blotting, Western, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, Epitopes, T-Lymphocyte immunology, Epitopes, T-Lymphocyte metabolism, Flow Cytometry, HLA-DP beta-Chains genetics, HLA-DP beta-Chains metabolism, Humans, Immunoglobulins immunology, Immunoglobulins metabolism, Immunotherapy methods, Interferon-gamma immunology, Interferon-gamma metabolism, Interleukin-4 immunology, Interleukin-4 metabolism, K562 Cells, Membrane Glycoproteins immunology, Membrane Glycoproteins metabolism, Molecular Sequence Data, Neoplasms immunology, Neoplasms metabolism, Neoplasms therapy, Phosphoproteins immunology, Survivin, Th1 Cells metabolism, Th2 Cells metabolism, Viral Matrix Proteins immunology, CD83 Antigen, Antigen-Presenting Cells immunology, HLA-DP beta-Chains immunology, Inhibitor of Apoptosis Proteins immunology, Th1 Cells immunology, Th2 Cells immunology
- Abstract
Purpose: In previous cancer vaccine clinical trials targeting survivin, induction of specific CD8(+) T-cell responses did not consistently lead to clinical responses. Considering the critical role of CD4(+) T-cell help in generating antitumor immunity, integration of anti-survivin CD4(+) T-cell responses may enhance the efficacy of anti-survivin cancer immunotherapy. Human leukocyte antigen (HLA)-DP4 is emerging as an attractive MHC target allele of CD4(+) T cell-mediated immunotherapy, because it is one of the most frequent HLA alleles in many ethnic groups. In this article, we aimed to elucidate DP4-restricted CD4(+) T-cell responses against survivin in cancer patients., Experimental Design: We generated a human cell-based artificial antigen-presenting cell (aAPC) expressing HLA-DP4, CD80, and CD83 and induced DP4-restricted antigen-specific CD4(+) T cells. The number, phenotype, effector function, and in vitro longevity of generated CD4(+) T cells were determined., Results: We first determined previously unknown DP4-restricted CD4(+) T-cell epitopes derived from cytomegalovirus pp65, to which sustained Th1-biased recall responses were induced in vitro by using DP4-aAPC. In contrast, DP4-aAPC induced in vitro both Th1 and Th2 long-lived anti-survivin CD4(+) T cells from cancer patients. Both survivin-specific Th1 and Th2 cells were able to recognize survivin-expressing tumors in a DP4-restricted manner. Neither survivin-specific interleukin 10 secreting Tr1 cells nor Th17 cells were induced by DP4-aAPC., Conclusions: DP4-restricted anti-survivin Th1 and Th2 immunity with sufficient functional avidity can be induced from cancer patients. The development of strategies to concurrently induce both CD4(+) and CD8(+) T-cell responses against survivin is warranted for optimal anti-survivin cancer immunotherapy., (©2011 AACR.)
- Published
- 2011
- Full Text
- View/download PDF
28. Mutations in the DDR2 kinase gene identify a novel therapeutic target in squamous cell lung cancer.
- Author
-
Hammerman PS, Sos ML, Ramos AH, Xu C, Dutt A, Zhou W, Brace LE, Woods BA, Lin W, Zhang J, Deng X, Lim SM, Heynck S, Peifer M, Simard JR, Lawrence MS, Onofrio RC, Salvesen HB, Seidel D, Zander T, Heuckmann JM, Soltermann A, Moch H, Koker M, Leenders F, Gabler F, Querings S, Ansén S, Brambilla E, Brambilla C, Lorimier P, Brustugun OT, Helland A, Petersen I, Clement JH, Groen H, Timens W, Sietsma H, Stoelben E, Wolf J, Beer DG, Tsao MS, Hanna M, Hatton C, Eck MJ, Janne PA, Johnson BE, Winckler W, Greulich H, Bass AJ, Cho J, Rauh D, Gray NS, Wong KK, Haura EB, Thomas RK, and Meyerson M
- Subjects
- Animals, Carcinoma, Non-Small-Cell Lung enzymology, Cell Transformation, Neoplastic drug effects, Cell Transformation, Neoplastic genetics, Dasatinib, Discoidin Domain Receptors, Erlotinib Hydrochloride, Humans, Lung Neoplasms enzymology, Mice, Mice, Nude, Mutation, NIH 3T3 Cells, Phosphorylation drug effects, Phosphorylation genetics, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Pyrimidines pharmacology, Pyrimidines therapeutic use, Quinazolines pharmacology, Quinazolines therapeutic use, Thiazoles pharmacology, Thiazoles therapeutic use, src-Family Kinases genetics, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung genetics, Lung Neoplasms drug therapy, Lung Neoplasms genetics, Receptor Protein-Tyrosine Kinases genetics, Receptors, Mitogen genetics
- Abstract
Unlabelled: While genomically targeted therapies have improved outcomes for patients with lung adenocarcinoma, little is known about the genomic alterations which drive squamous cell lung cancer. Sanger sequencing of the tyrosine kinome identified mutations in the DDR2 kinase gene in 3.8% of squamous cell lung cancers and cell lines. Squamous lung cancer cell lines harboring DDR2 mutations were selectively killed by knock-down of DDR2 by RNAi or by treatment with the multi-targeted kinase inhibitor dasatinib. Tumors established from a DDR2 mutant cell line were sensitive to dasatinib in xenograft models. Expression of mutated DDR2 led to cellular transformation which was blocked by dasatinib. A squamous cell lung cancer patient with a response to dasatinib and erlotinib treatment harbored a DDR2 kinase domain mutation. These data suggest that gain-of-function mutations in DDR2 are important oncogenic events and are amenable to therapy with dasatinib. As dasatinib is already approved for use, these findings could be rapidly translated into clinical trials., Significance: DDR2 mutations are present in 4% of lung SCCs, and DDR2 mutations are associated with sensitivity to dasatinib. These findings provide a rationale for designing clinical trials with the FDA-approved drug dasatinib in patients with lung SCCs.
- Published
- 2011
- Full Text
- View/download PDF
29. Blood-based gene expression signatures in non-small cell lung cancer.
- Author
-
Zander T, Hofmann A, Staratschek-Jox A, Classen S, Debey-Pascher S, Maisel D, Ansén S, Hahn M, Beyer M, Thomas RK, Gathof B, Mauch C, Delank KS, Engel-Riedel W, Wichmann HE, Stoelben E, Schultze JL, and Wolf J
- Subjects
- Adult, Aged, Algorithms, Carcinoma, Non-Small-Cell Lung blood, Carcinoma, Non-Small-Cell Lung classification, Case-Control Studies, Female, Gene Expression Regulation, Neoplastic, Humans, Lung Neoplasms blood, Lung Neoplasms classification, Male, Middle Aged, Oligonucleotide Array Sequence Analysis, Carcinoma, Non-Small-Cell Lung genetics, Gene Expression Profiling, Leukocytes, Mononuclear metabolism, Lung Neoplasms genetics
- Abstract
Purpose: Blood-based surrogate markers would be attractive biomarkers for early detection, diagnosis, prognosis, and prediction of therapeutic outcome in cancer. Disease-associated gene expression signatures in peripheral blood mononuclear cells (PBMC) have been described for several cancer types. However, RNA-stabilized whole blood-based technologies would be clinically more applicable and robust. We evaluated the applicability of whole blood-based gene expression profiling for the detection of non-small cell lung cancer (NSCLC)., Experimental Design: Expression profiles were generated from PAXgene-stabilized blood samples from three independent groups consisting of NSCLC cases and controls (n = 77, 54, and 102), using the Illumina WG6-VS2 system., Results: Several genes are consistently differentially expressed in whole blood of NSCLC patients and controls. These expression profiles were used to build a diagnostic classifier for NSCLC, which was validated in an independent validation set of NSCLC patients (stages I-IV) and hospital-based controls. The area under the receiver operator curve was calculated to be 0.824 (P < 0.001). In a further independent dataset of stage I NSCLC patients and healthy controls the AUC was 0.977 (P < 0.001). Specificity of the classifier was validated by permutation analysis in both validation cohorts. Genes within the classifier are enriched in immune-associated genes and show specificity for NSCLC., Conclusions: Our results show that gene expression profiles of whole blood allow for detection of manifest NSCLC. These results prompt further development of gene expression-based biomarker tests in peripheral blood for the diagnosis and early detection of NSCLC., (©2011 AACR.)
- Published
- 2011
- Full Text
- View/download PDF
30. Benchmarking of mutation diagnostics in clinical lung cancer specimens.
- Author
-
Querings S, Altmüller J, Ansén S, Zander T, Seidel D, Gabler F, Peifer M, Markert E, Stemshorn K, Timmermann B, Saal B, Klose S, Ernestus K, Scheffler M, Engel-Riedel W, Stoelben E, Brambilla E, Wolf J, Nürnberg P, and Thomas RK
- Subjects
- Adult, Aged, Base Sequence, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung pathology, ErbB Receptors genetics, Exons genetics, Female, Genome, Human genetics, Humans, Lung Neoplasms drug therapy, Lung Neoplasms pathology, Male, Middle Aged, Molecular Sequence Data, Treatment Outcome, Benchmarking, DNA Mutational Analysis methods, DNA Mutational Analysis standards, Lung Neoplasms genetics, Mutation genetics
- Abstract
Treatment of EGFR-mutant non-small cell lung cancer patients with the tyrosine kinase inhibitors erlotinib or gefitinib results in high response rates and prolonged progression-free survival. Despite the development of sensitive mutation detection approaches, a thorough validation of these in a clinical setting has so far been lacking. We performed, in a clinical setting, a systematic validation of dideoxy 'Sanger' sequencing and pyrosequencing against massively parallel sequencing as one of the most sensitive mutation detection technologies available. Mutational annotation of clinical lung tumor samples revealed that of all patients with a confirmed response to EGFR inhibition, only massively parallel sequencing detected all relevant mutations. By contrast, dideoxy sequencing missed four responders and pyrosequencing missed two responders, indicating a dramatic lack of sensitivity of dideoxy sequencing, which is widely applied for this purpose. Furthermore, precise quantification of mutant alleles revealed a low correlation (r(2) = 0.27) of histopathological estimates of tumor content and frequency of mutant alleles, thereby questioning the use of histopathology for stratification of specimens for individual analytical procedures. Our results suggest that enhanced analytical sensitivity is critically required to correctly identify patients responding to EGFR inhibition. More broadly, our results emphasize the need for thorough evaluation of all mutation detection approaches against massively parallel sequencing as a prerequisite for any clinical implementation.
- Published
- 2011
- Full Text
- View/download PDF
31. Frequent and focal FGFR1 amplification associates with therapeutically tractable FGFR1 dependency in squamous cell lung cancer.
- Author
-
Weiss J, Sos ML, Seidel D, Peifer M, Zander T, Heuckmann JM, Ullrich RT, Menon R, Maier S, Soltermann A, Moch H, Wagener P, Fischer F, Heynck S, Koker M, Schöttle J, Leenders F, Gabler F, Dabow I, Querings S, Heukamp LC, Balke-Want H, Ansén S, Rauh D, Baessmann I, Altmüller J, Wainer Z, Conron M, Wright G, Russell P, Solomon B, Brambilla E, Brambilla C, Lorimier P, Sollberg S, Brustugun OT, Engel-Riedel W, Ludwig C, Petersen I, Sänger J, Clement J, Groen H, Timens W, Sietsma H, Thunnissen E, Smit E, Heideman D, Cappuzzo F, Ligorio C, Damiani S, Hallek M, Beroukhim R, Pao W, Klebl B, Baumann M, Buettner R, Ernestus K, Stoelben E, Wolf J, Nürnberg P, Perner S, and Thomas RK
- Subjects
- Adenocarcinoma drug therapy, Adenocarcinoma genetics, Adenocarcinoma metabolism, Adenocarcinoma of Lung, Animals, Apoptosis genetics, Apoptosis physiology, Blotting, Western, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung metabolism, Cell Line, Enzyme Inhibitors therapeutic use, Gene Expression Regulation, Neoplastic genetics, Humans, Lung Neoplasms drug therapy, Male, Mice, Mice, Nude, Pyrimidines therapeutic use, RNA Interference, Receptor, Fibroblast Growth Factor, Type 1 antagonists & inhibitors, Receptor, Fibroblast Growth Factor, Type 1 genetics, Xenograft Model Antitumor Assays, Lung Neoplasms genetics, Lung Neoplasms metabolism, Receptor, Fibroblast Growth Factor, Type 1 metabolism
- Abstract
Lung cancer remains one of the leading causes of cancer-related death in developed countries. Although lung adenocarcinomas with EGFR mutations or EML4-ALK fusions respond to treatment by epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) inhibition, respectively, squamous cell lung cancer currently lacks therapeutically exploitable genetic alterations. We conducted a systematic search in a set of 232 lung cancer specimens for genetic alterations that were therapeutically amenable and then performed high-resolution gene copy number analyses. We identified frequent and focal fibroblast growth factor receptor 1 (FGFR1) amplification in squamous cell lung cancer (n = 155), but not in other lung cancer subtypes, and, by fluorescence in situ hybridization, confirmed the presence of FGFR1 amplifications in an independent cohort of squamous cell lung cancer samples (22% of cases). Using cell-based screening with the FGFR inhibitor PD173074 in a large (n = 83) panel of lung cancer cell lines, we demonstrated that this compound inhibited growth and induced apoptosis specifically in those lung cancer cells carrying amplified FGFR1. We validated the FGFR1 dependence of FGFR1-amplified cell lines by FGFR1 knockdown and by ectopic expression of an FGFR1-resistant allele (FGFR1(V561M)), which rescued FGFR1-amplified cells from PD173074-mediated cytotoxicity. Finally, we showed that inhibition of FGFR1 with a small molecule led to significant tumor shrinkage in vivo. Thus, focal FGFR1 amplification is common in squamous cell lung cancer and associated with tumor growth and survival, suggesting that FGFR inhibitors may be a viable therapeutic option in this cohort of patients.
- Published
- 2010
- Full Text
- View/download PDF
32. A panel of human cell-based artificial APC enables the expansion of long-lived antigen-specific CD4+ T cells restricted by prevalent HLA-DR alleles.
- Author
-
Butler MO, Ansén S, Tanaka M, Imataki O, Berezovskaya A, Mooney MM, Metzler G, Milstein MI, Nadler LM, and Hirano N
- Subjects
- CD8-Positive T-Lymphocytes immunology, Cell Line, Humans, Alleles, Antigen-Presenting Cells immunology, CD4-Positive T-Lymphocytes immunology, HLA-DR Antigens immunology
- Abstract
Many preclinical experiments have attested to the critical role of CD4(+) T cell help in CD8(+) cytotoxic T lymphocyte (CTL)-mediated immunity. Recent clinical trials have demonstrated that reinfusion of CD4(+) T cells can induce responses in infectious diseases and cancer. However, few standardized and versatile systems exist to expand antigen-specific CD4(+) T(h) for clinical use. K562 is a human erythroleukemic cell line, which lacks expression of HLA class I and class II, invariant chain and HLA-DM but expresses adhesion molecules such as intercellular adhesion molecule-1 and leukocyte function-associated antigen-3. With this unique immunologic phenotype, K562 has been tested in clinical trials of cancer immunotherapy. Previously, we created a K562-based artificial antigen-presenting cell (aAPC) that generates ex vivo long-lived HLA-A2-restricted CD8(+) CTL with a central/effector memory phenotype armed with potent effector function. We successfully generated a clinical version of this aAPC and conducted a clinical trial where large numbers of anti-tumor CTL are reinfused to cancer patients. In this article, we shifted focus to CD4(+) T cells and developed a panel of novel K562-derived aAPC, where each expresses a different single HLA-DR allele, invariant chain, HLA-DM, CD80, CD83 and CD64; takes up soluble protein by endocytosis and processes and presents CD4(+) T-cell peptides. Using this aAPC, we were able to determine novel DR-restricted CD4(+) T-cell epitopes and expand long-lived CD4(+) T-cells specific for multiple antigens without growing bystander Foxp3(+) regulatory T cells. Our results suggest that K562-based aAPC may serve as a translatable platform to generate both antigen-specific CD8(+) CTL and CD4(+) T(h).
- Published
- 2010
- Full Text
- View/download PDF
33. Osteoblastic response in patients with non-small cell lung cancer with activating EGFR Mutations and bone metastases during treatment with EGFR kinase inhibitors.
- Author
-
Ansén S, Bangard C, Querings S, Gabler F, Scheffler M, Seidel D, Saal B, Zander T, Nogová L, Töpelt K, Markert E, Stoelben E, Ernestus K, Thomas RK, and Wolf J
- Subjects
- Adenocarcinoma drug therapy, Adenocarcinoma genetics, Adenocarcinoma pathology, Bone Neoplasms genetics, Bone Neoplasms secondary, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung pathology, ErbB Receptors antagonists & inhibitors, Female, Humans, Lung Neoplasms genetics, Lung Neoplasms pathology, Middle Aged, Prognosis, Survival Rate, Bone Neoplasms drug therapy, Carcinoma, Non-Small-Cell Lung drug therapy, ErbB Receptors genetics, Lung Neoplasms drug therapy, Mutation genetics, Osteoblasts drug effects, Protein Kinase Inhibitors therapeutic use
- Published
- 2010
- Full Text
- View/download PDF
34. Identifying genotype-dependent efficacy of single and combined PI3K- and MAPK-pathway inhibition in cancer.
- Author
-
Sos ML, Fischer S, Ullrich R, Peifer M, Heuckmann JM, Koker M, Heynck S, Stückrath I, Weiss J, Fischer F, Michel K, Goel A, Regales L, Politi KA, Perera S, Getlik M, Heukamp LC, Ansén S, Zander T, Beroukhim R, Kashkar H, Shokat KM, Sellers WR, Rauh D, Orr C, Hoeflich KP, Friedman L, Wong KK, Pao W, and Thomas RK
- Subjects
- Apoptosis drug effects, Cell Line, Tumor, Genotype, Humans, Neoplasms enzymology, Neoplasms pathology, Phosphoinositide-3 Kinase Inhibitors, MAP Kinase Signaling System drug effects, Neoplasms drug therapy, Neoplasms genetics, Phosphatidylinositol 3-Kinases genetics, Protein Kinase Inhibitors pharmacology
- Abstract
In cancer, genetically activated proto-oncogenes often induce "upstream" dependency on the activity of the mutant oncoprotein. Therapeutic inhibition of these activated oncoproteins can induce massive apoptosis of tumor cells, leading to sometimes dramatic tumor regressions in patients. The PI3K and MAPK signaling pathways are central regulators of oncogenic transformation and tumor maintenance. We hypothesized that upstream dependency engages either one of these pathways preferentially to induce "downstream" dependency. Therefore, we analyzed whether downstream pathway dependency segregates by genetic aberrations upstream in lung cancer cell lines. Here, we show by systematically linking drug response to genomic aberrations in non-small-cell lung cancer, as well as in cell lines of other tumor types and in a series of in vivo cancer models, that tumors with genetically activated receptor tyrosine kinases depend on PI3K signaling, whereas tumors with mutations in the RAS/RAF axis depend on MAPK signaling. However, efficacy of downstream pathway inhibition was limited by release of negative feedback loops on the reciprocal pathway. By contrast, combined blockade of both pathways was able to overcome the reciprocal pathway activation induced by inhibitor-mediated release of negative feedback loops and resulted in a significant increase in apoptosis and tumor shrinkage. Thus, by using a systematic chemo-genomics approach, we identify genetic lesions connected to PI3K and MAPK pathway activation and provide a rationale for combined inhibition of both pathways. Our findings may have implications for patient stratification in clinical trials.
- Published
- 2009
- Full Text
- View/download PDF
35. Dissociation of its opposing immunologic effects is critical for the optimization of antitumor CD8+ T-cell responses induced by interleukin 21.
- Author
-
Ansén S, Butler MO, Berezovskaya A, Murray AP, Stevenson K, Nadler LM, and Hirano N
- Subjects
- Antigen-Presenting Cells metabolism, Cell Proliferation, Cytokines metabolism, Dose-Response Relationship, Drug, Histocompatibility Antigens Class I chemistry, Humans, Immunotherapy, Adoptive methods, Phenotype, Receptors, Interleukin-21 genetics, T-Lymphocytes immunology, Time Factors, Treatment Outcome, CD8-Positive T-Lymphocytes cytology, CD8-Positive T-Lymphocytes immunology, Dendritic Cells cytology, Interleukins metabolism, Receptors, Interleukin-21 metabolism
- Abstract
Purpose: Interleukin 21 (IL-21) is a promising new cytokine, which is undergoing clinical testing as an anticancer agent. Although IL-21 provides potent stimulation of CD8(+) T cells, it has also been suggested that IL-21 is immunosuppressive by counteracting the maturation of dendritic cells. The dissociation of these two opposing effects may enhance the utility of IL-21 as an immunotherapeutic. In this study, we used a cell-based artificial antigen-presenting cell (aAPC) lacking a functional IL-21 receptor (IL-21R) to investigate the immunostimulatory properties of IL-21., Experimental Design: The immunosuppressive activity of IL-21 was studied using human IL-21R(+) dendritic cells. Antigen-specific CD8(+) T cells stimulated with human cell-based IL-21R(-)aAPC were used to isolate the T-cell immunostimulatory effects of IL-21. The functional outcomes, including phenotype, cytokine production, proliferation, and cytotoxicity were evaluated., Results: IL-21 limits the immune response by maintaining immunologically immature dendritic cells. However, stimulation of CD8(+) T cells with IL-21R(-) aAPC, which secrete IL-21, results in significant expansion. Although priming in the presence of IL-21 temporarily modulated the T-cell phenotype, chronic stimulation abrogated these differences. Importantly, exposure to IL-21 during restimulation promoted the enrichment and expansion of antigen-specific CD8(+) T cells that maintained IL-2 secretion and gained enhanced IFN-gamma secretion. Tumor antigen-specific CTL generated in the presence of IL-21 recognized tumor cells efficiently, demonstrating potent effector functions., Conclusions: IL-21 induces opposing effects on antigen-presenting cells and CD8(+) T cells. Strategic application of IL-21 is required to induce optimal clinical effects and may enable the generation of large numbers of highly avid tumor-specific CTL for adoptive immunotherapy.
- Published
- 2008
- Full Text
- View/download PDF
36. Long-lived antitumor CD8+ lymphocytes for adoptive therapy generated using an artificial antigen-presenting cell.
- Author
-
Butler MO, Lee JS, Ansén S, Neuberg D, Hodi FS, Murray AP, Drury L, Berezovskaya A, Mulligan RC, Nadler LM, and Hirano N
- Subjects
- Antigen Presentation immunology, Antigen-Presenting Cells immunology, Antigens, Neoplasm metabolism, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, Cell Proliferation drug effects, Half-Life, Humans, Interferon-gamma metabolism, Interleukin-15 pharmacology, K562 Cells, MART-1 Antigen, Melanoma immunology, Melanoma metabolism, Melanoma pathology, Neoplasm Proteins metabolism, Tumor Cells, Cultured, Antigen-Presenting Cells transplantation, Biomimetics methods, CD8-Positive T-Lymphocytes transplantation, Immunotherapy, Adoptive, Melanoma therapy
- Abstract
Purpose: Antitumor lymphocytes can be generated ex vivo unencumbered by immunoregulation found in vivo. Adoptive transfer of these cells is a promising therapeutic modality that could establish long-term antitumor immunity. However, the widespread use of adoptive therapy has been hampered by the difficulty of consistently generating potent antitumor lymphocytes in a timely manner for every patient. To overcome this, we sought to establish a clinical grade culture system that can reproducibly generate antigen-specific cytotoxic T lymphocytes (CTL)., Experimental Design: We created an off-the-shelf, standardized, and renewable artificial antigen-presenting cell (aAPC) line that coexpresses HLA class I, CD54, CD58, CD80, and the dendritic cell maturation marker CD83. We tested the ability of aAPC to generate tumor antigen-specific CTL under optimal culture conditions. The number, phenotype, effector function, and in vitro longevity of generated CTL were determined., Results: Stimulation of CD8(+) T cells with peptide-pulsed aAPC generated large numbers of functional CTL that recognized a variety of tumor antigens. These CTLs, which possess a phenotype consistent with in vivo persistence, survived ex vivo for prolonged periods of time. Clinical grade aAPC(33), produced under current Good Manufacturing Practices guidelines, generated sufficient numbers of CTL within a short period of time. These CTL specifically lysed a variety of melanoma tumor lines naturally expressing a target melanoma antigen. Furthermore, antitumor CTL were easily generated in all melanoma patients examined., Conclusions: With clinical grade aAPC(33) in hand, we are now poised for clinical translation of ex vivo generated antitumor CTL for adoptive cell transfer.
- Published
- 2007
- Full Text
- View/download PDF
37. Identification of an immunogenic CD8+ T-cell epitope derived from gamma-globin, a putative tumor-associated antigen for juvenile myelomonocytic leukemia.
- Author
-
Hirano N, Butler MO, Xia Z, Berezovskaya A, Murray AP, Ansén S, Kojima S, and Nadler LM
- Subjects
- Antigen-Presenting Cells immunology, Antigens, Neoplasm genetics, Child, Epitopes isolation & purification, HLA-A2 Antigen metabolism, Humans, In Vitro Techniques, Leukemia, Myelomonocytic, Acute genetics, Antigens, Neoplasm isolation & purification, CD8-Positive T-Lymphocytes immunology, Globins immunology, Leukemia, Myelomonocytic, Acute immunology
- Abstract
Juvenile myelomonocytic leukemia (JMML) is a rare clonal myeloproliferative disorder. Although allogeneic stem cell transplantation can induce long-term remissions, relapse rates remain high and innovative approaches are needed. Since donor lymphocyte infusions have clinical activity in JMML, T-cell-mediated immunotherapy could provide a nonredundant treatment approach to compliment current therapies. Gamma-globin, an oncofetal protein overexpressed by clonogenic JMML cells, may serve as a target of an antitumor immune response. We predicted 5 gamma-globin-derived peptides as potential human leukocyte antigen (HLA)-A2 restricted cytotoxic T lymphocyte (CTL) epitopes and showed that 4 (g031, g071, g105, and g106) bind A2 molecules in vitro. Using an artificial antigen-presenting cell (aAPC) that can process both the N- and C-termini of endogenously expressed proteins, we biochemically confirmed that g105 is naturally processed and presented by cell surface A2. Furthermore, g105-specific CD8(+) CTLs generated from A2-positive healthy donors were able to specifically cytolyze gamma-globin(+), but not gamma-globin(-) JMML cells in an A2-restricted manner. These results suggest that this aAPC-based approach enables the biochemical identification of CD8(+) T-cell epitopes that are processed and presented by intact cells, and that CTL immunotherapy of JMML could be directed against the gamma-globin-derived epitope g105.
- Published
- 2006
- Full Text
- View/download PDF
38. Efficient presentation of naturally processed HLA class I peptides by artificial antigen-presenting cells for the generation of effective antitumor responses.
- Author
-
Hirano N, Butler MO, Xia Z, Berezovskaya A, Murray AP, Ansén S, and Nadler LM
- Subjects
- Antigens, Neoplasm, HLA-A2 Antigen, Humans, Immunotherapy methods, MART-1 Antigen, Neoplasm Proteins immunology, Neoplasms immunology, Neoplasms therapy, Peptide Fragments, Pregnancy Proteins immunology, T-Lymphocytes, Cytotoxic immunology, Antigen Presentation, Antigen-Presenting Cells immunology, HLA-A Antigens immunology
- Abstract
Appropriate presentation of tumor-associated antigens (TAA) by antigen-presenting cells (APC) is required for the development of clinically relevant antitumor T-cell responses. One common approach, which uses APC pulsed with synthetic peptides, can sometimes generate ineffective immune responses. This failure may, in part, be attributed to the formation of HLA/synthetic pulsed peptide complexes that possess different conformations compared with those of endogenously presented peptides. In addition, endogenous peptides may undergo post-translational modifications, which do not occur with synthetic peptides. Because our goal is to induce immunity that can recognize TAA that are endogenously presented by tumors, we designed an APC that would not only express the required immunoaccessory molecules but also naturally process and present target antigenic peptides. In this study, we generated an artificial APC (aAPC) that can endogenously present any chosen HLA-A*0201 (A2)-restricted peptide by processing a fusion protein that contains a unique "LTK" sequence linked to the antigenic peptide. Proteasome-dependent processing is so effective that the presented peptide can be directly eluted from the cell surface and identified by biochemical methods. Furthermore, we found that aAPC, engineered to endogenously present peptide derived from the melanoma antigen MART1, can be used to prime and expand antitumor CTL that target MART1-expressing tumor cells in a HLA-A2-restricted manner. Our engineered aAPC could serve as an "off-the-shelf" APC designed to constitutively express class I-restricted TAA peptides and could be used to generate effective T-cell responses to treat human disease.
- Published
- 2006
- Full Text
- View/download PDF
39. Engagement of CD83 ligand induces prolonged expansion of CD8+ T cells and preferential enrichment for antigen specificity.
- Author
-
Hirano N, Butler MO, Xia Z, Ansén S, von Bergwelt-Baildon MS, Neuberg D, Freeman GJ, and Nadler LM
- Subjects
- CD28 Antigens immunology, Cloning, Molecular, Humans, Jurkat Cells, Lymphocyte Activation, CD83 Antigen, Antigens, CD immunology, CD8-Positive T-Lymphocytes immunology, Immunoglobulins immunology, Membrane Glycoproteins immunology
- Abstract
Following T-cell receptor and CD28 signaling, CD8+ T cells express a receptor for CD83, a molecule up-regulated on functionally mature dendritic cells. Although this expression pattern suggests that CD83 is involved in adaptive immunity, little is known about its function in the periphery, and the existence of its ligand on T cells is controversial. We demonstrate that the engagement of the CD83 ligand (CD83L) preferentially enriches and significantly amplifies the number of antigen-specific CD8+ T cells. Coengagement of the T-cell receptor, CD28, and CD83L supports priming of naive CD8+ T cells that retain antigen specificity and cytotoxic function for more than 6 months. Therefore, engagement of the CD83L provides a unique signal to activated CD8+ T cells that could be exploited to generate long-lived antigen-specific cytotoxic T cells for the treatment of cancer and infection.
- Published
- 2006
- Full Text
- View/download PDF
40. Gene expression profiling identifies BAX-delta as a novel tumor antigen in acute lymphoblastic leukemia.
- Author
-
Maia S, Haining WN, Ansén S, Xia Z, Armstrong SA, Seth NP, Ghia P, den Boer ML, Pieters R, Sallan SE, Nadler LM, and Cardoso AA
- Subjects
- Adolescent, Amino Acid Sequence, Antigens, Neoplasm biosynthesis, B-Lymphocytes immunology, Cell Line, Tumor, Child, Child, Preschool, Epitopes, T-Lymphocyte immunology, Gene Expression Profiling, HLA-A Antigens immunology, HLA-A2 Antigen, HLA-DR Antigens immunology, Humans, Infant, Lymphocyte Activation immunology, Molecular Sequence Data, T-Lymphocytes, Cytotoxic immunology, bcl-2-Associated X Protein biosynthesis, Antigens, Neoplasm genetics, Antigens, Neoplasm immunology, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma immunology, bcl-2-Associated X Protein genetics, bcl-2-Associated X Protein immunology
- Abstract
The identification of new tumor-associated antigens (TAA) is critical for the development of effective immunotherapeutic strategies, particularly in diseases like B-cell acute lymphoblastic leukemia (B-ALL), where few target epitopes are known. To accelerate the identification of novel TAA in B-ALL, we used a combination of expression profiling and reverse immunology. We compared gene expression profiles of primary B-ALL cells with their normal counterparts, B-cell precursors. Genes differentially expressed by B-ALL cells included many previously identified as TAA in other malignancies. Within this set of overexpressed genes, we focused on those that may be functionally important to the cancer cell. The apoptosis-related molecule, BAX, was highly correlated with the ALL class distinction. Therefore, we evaluated BAX and its isoforms as potential TAA. Peptides from the isoform BAX-delta bound with high affinity to HLA-A*0201 and HLA-DR1. CD8+ CTLs specific for BAX-delta epitopes or their heteroclitic peptides could be expanded from normal donors. BAX-delta-specific T cells lysed peptide-pulsed targets and BAX-delta-expressing leukemia cells in a MHC-restricted fashion. Moreover, primary B-ALL cells were recognized by BAX-delta-specific CTL, indicating that this antigen is naturally processed and presented by tumor cells. This study suggests that (a) BAX-delta may serve as a widely expressed TAA in B-ALL and (b) gene expression profiling can be a generalizable tool to identify immunologic targets for cancer immunotherapy.
- Published
- 2005
- Full Text
- View/download PDF
41. Phase I/II combined chemoimmunotherapy with carcinoembryonic antigen-derived HLA-A2-restricted CAP-1 peptide and irinotecan, 5-fluorouracil, and leucovorin in patients with primary metastatic colorectal cancer.
- Author
-
Weihrauch MR, Ansén S, Jurkiewicz E, Geisen C, Xia Z, Anderson KS, Gracien E, Schmidt M, Wittig B, Diehl V, Wolf J, Bohlen H, and Nadler LM
- Subjects
- Adult, Aged, CD3 Complex analysis, CD4 Antigens analysis, CD8 Antigens analysis, Camptothecin administration & dosage, Camptothecin analogs & derivatives, Carcinoembryonic Antigen blood, Colorectal Neoplasms immunology, Colorectal Neoplasms pathology, Combined Modality Therapy, Dose-Response Relationship, Drug, Female, Flow Cytometry, Fluorouracil administration & dosage, Granulocyte-Macrophage Colony-Stimulating Factor administration & dosage, Humans, Immunodominant Epitopes immunology, Immunotherapy methods, Interferon-gamma metabolism, Interleukin-2 administration & dosage, Irinotecan, Leucovorin administration & dosage, Liver Neoplasms immunology, Liver Neoplasms secondary, Liver Neoplasms therapy, Male, Middle Aged, Oligopeptides administration & dosage, Oligopeptides immunology, Prospective Studies, T-Lymphocytes cytology, T-Lymphocytes drug effects, T-Lymphocytes immunology, Treatment Outcome, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Carcinoembryonic Antigen immunology, Colorectal Neoplasms therapy, HLA-A2 Antigen immunology, Oligopeptides therapeutic use
- Abstract
Purpose: We conducted a phase I/II randomized trial to evaluate the clinical and immunologic effect of chemotherapy combined with vaccination in primary metastatic colorectal cancer patients with a carcinoembryonic antigen-derived peptide in the setting of adjuvants granulocyte macrophage colony-stimulating factor, CpG-containing DNA molecules (dSLIM), and dendritic cells., Experimental Design: HLA-A2-positive patients with confirmed newly diagnosed metastatic colorectal cancer and elevated serum carcinoembryonic antigen (CEA) were randomized to receive three cycles of standard chemotherapy (irinotecan/high-dose 5-fluorouracil/leucovorin) and vaccinations with CEA-derived CAP-1 peptide admixed with different adjuvants [CAP-1/granulocyte macrophage colony-stimulating factor/interleukin-2 (IL-2), CAP-1/dSLIM/IL-2, and CAP-1/IL-2]. After completion of chemotherapy, patients received weekly vaccinations until progression of disease. Immune assessment was done at baseline and after three cycles of combined chemoimmunotherapy. HLA-A2 tetramers complexed with the peptides CAP-1, human T-cell lymphotrophic virus type I TAX, cytomegalovirus (CMV) pp65, and EBV BMLF-1 were used for phenotypic immune assessment. IFN-gamma intracellular cytokine assays were done to evaluate CTL reactivity., Results: Seventeen metastatic patients were recruited, of whom 12 completed three cycles. Therapy resulted in five complete response, one partial response, five stable disease, and six progressive disease. Six grade 1 local skin reactions and one mild systemic reaction to vaccination treatment were observed. Overall survival after a median observation time of 29 months was 17 months with a survival rate of 35% (6 of 17) at that time. Eight patients (47%) showed elevation of CAP-1-specific CTLs. Neither of the adjuvants provided superiority in eliciting CAP-1-specific immune responses. During three cycles of chemotherapy, EBV/CMV recall antigen-specific CD8+ cells decreased by an average 14%., Conclusions: The presented chemoimmunotherapy is a feasible and safe combination therapy with clinical and immunologic efficacy. Despite concurrent chemotherapy, increases in CAP-1-specific T cells were observed in 47% of patients after vaccination.
- Published
- 2005
- Full Text
- View/download PDF
42. Immunoglobulin gene segment usage, location and immunogenicity in mutated and unmutated chronic lymphocytic leukaemia.
- Author
-
Mauerer K, Zahrieh D, Gorgun G, Li A, Zhou J, Ansén S, Rassenti LZ, and Gribben JG
- Subjects
- Amino Acid Sequence, DNA Mutational Analysis, Epitopes, HLA-DR3 Antigen genetics, HLA-DR3 Antigen immunology, Histocompatibility Antigens Class I immunology, Histocompatibility Antigens Class II immunology, Humans, Immunoglobulin Heavy Chains immunology, Immunoglobulin Variable Region immunology, Leukemia, Lymphocytic, Chronic, B-Cell immunology, Molecular Sequence Data, Prognosis, Sequence Alignment, Gene Rearrangement, B-Lymphocyte, Heavy Chain, Genes, Immunoglobulin, Immunoglobulin Heavy Chains genetics, Immunoglobulin Variable Region genetics, Leukemia, Lymphocytic, Chronic, B-Cell genetics
- Abstract
The mutational status of the variable region of the immunoglobulin heavy chain gene (IgV(H)) is an important prognostic marker in B-cell chronic lymphocytic leukaemia (B-CLL), with mutated patients having improved outcome. To examine the impact of mutational status on V(H), D(H), and J(H) gene segment location and immunogenicity, we analysed 375 IgH sequences from 356 patients with B-CLL. Although V(H) and D(H) gene usage was different in mutated compared to unmutated patients, there was no impact of gene location on frequency of use or clinical outcome. Surprisingly, somatic mutations did not increase the immunogenicity of the Ig, as assessed by predicted binding affinity of Ig-derived peptides to major histocompatibility Class I and Class II molecules. Even excluding patients using V(H)1-69, cases using the V(H)1 gene family had a poor outcome. Both mutated and unmutated CLL patients demonstrated evidence of antigen selection. The worst outcome was seen in the subset of 14 unmutated patients with similar HCDR3 amino acid sequence using V(H)1-69, D(H)3-3 and J(H)6, suggesting an antigen-driven process modulating the clinical course.
- Published
- 2005
- Full Text
- View/download PDF
43. [New therapeutic strategies for Hodgkin lymphoma in cooperation of radiation oncology and medical oncology].
- Author
-
Eich HT, Müller RP, Ansén S, Josting A, Engert A, Hansemann K, Pfistner B, Wolf J, Willich N, and Diehl V
- Subjects
- Chemotherapy, Adjuvant, Combined Modality Therapy, Dose-Response Relationship, Drug, Drug Administration Schedule, Follow-Up Studies, Germany, Hodgkin Disease mortality, Hodgkin Disease pathology, Humans, Neoplasm Recurrence, Local drug therapy, Neoplasm Recurrence, Local mortality, Neoplasm Recurrence, Local pathology, Neoplasm Recurrence, Local radiotherapy, Neoplasm Staging, Radiotherapy Dosage, Radiotherapy Planning, Computer-Assisted, Radiotherapy, Adjuvant, Tomography, X-Ray Computed, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Hodgkin Disease drug therapy, Hodgkin Disease radiotherapy, Patient Care Team
- Abstract
Background: Between 1984 and 2002 more than 9500 pts. were enrolled in the multicentric randomized trials of the German Hodgkin Study Group (GHSG) and are evaluable for response, survival, recurrences, and toxicities. Actually the GHSG evaluates the efficacy of risk-adapted therapy composed of polychemotherapy (CT) and radiotherapy (RT). An extensive RT quality assurance program has been practiced during the study generations and will be continued., Patients and Methods: The 4th study generation (1998-2002) includes the following trials: In the HD10 trial (early stages) 4x ABVD are tested against 2x ABVD followed by 20 Gy Involved Field (IF)-RT vs. 30 Gy IF-RT (4 arms). In order to optimize CT-regimen and IF-RT dose for pts. with intermediate stage, the HD11 trial compares 4x ABVD with 4x BEACOPP baseline followed by 20 Gy IF-RT vs. 30 Gy IF-RT in a 4 arm design. Concerning advanced stages (HD12), the BEACOPP regimen is to be optimized and the necessity of additive RT is tested. The standard arm (8x BEACOPP escalated) is compared with the toxicity reduced arm (4x BEACOPP escalated + 4x BEACOPP baseline) followed by 30 Gy RT on initial bulky disease and/or residual tumor vs. no RT (4 arms)., Results: Interim results (without arm comparisons) with a median follow-up of 18 months for HD10 and HD11 and 20 months for HD12 are as follows: Freedom from Treatment Failure (FFTF) at 18 months is in the HD10 trial (390 pts.) 96.4%, in the HD11 trial (480 pts.) 91.5% and in the HD12 trial (550 pts.) 90.2%.The overall survival (OS) at 18 months is in HD10 98.2%, in HD11 98.5%, in HD12 93.5%. In HD10, HD11 and HD12 respectively, 1.8%, 1.9% and 2.5% of pts. died and 1.0%, 2.5% and 2.2% suffered early progression. CONCLUSION/FURTHER STRATEGY: In order to reduce the relapse rate and toxicity and to improve the quality of life, the new HD13 trial for early stages (Fig. 1a) compares 2x ABVD, 2x ABV, 2x AVD und 2x AV, each followed by 30 Gy IF-RT. For the intermediate stages, the FFTF rate should be improved by intensifying the standard regimen. Therefore the new trial HD14 (Fig. 1b) compares 4x ABVD with 2x BEACOPP escalated + 2x ABVD, each followed by 30 Gy IF-RT. In the new trial for advanced stages HD15 (Fig. 1c),the FFTF/OS rates are to be maintained and the quality of life to be improved. 8x BEACOPP escalated, 6x BEACOPP escalated and 8x BEACOPP baseline with shortened 14-day cycle are compared. The use of PET to decide on additive RT will also be investigated.
- Published
- 2003
44. [Current therapeutic strategies in Hodgkin lymphoma].
- Author
-
Ansén S, Sieber M, Wingbermühle K, Josting A, and Diehl V
- Subjects
- Chemotherapy, Adjuvant, Combined Modality Therapy, Critical Pathways, Hodgkin Disease drug therapy, Hodgkin Disease pathology, Humans, Neoplasm Staging, Randomized Controlled Trials as Topic, Antineoplastic Combined Chemotherapy Protocols, Hodgkin Disease radiotherapy
- Published
- 2002
- Full Text
- View/download PDF
45. Thoracic positron emission tomography using 18F-fluorodeoxyglucose for the evaluation of residual mediastinal Hodgkin disease.
- Author
-
Weihrauch MR, Re D, Scheidhauer K, Ansén S, Dietlein M, Bischoff S, Bohlen H, Wolf J, Schicha H, Diehl V, and Tesch H
- Subjects
- Antineoplastic Combined Chemotherapy Protocols administration & dosage, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Bleomycin administration & dosage, Carmustine administration & dosage, Combined Modality Therapy, Cyclophosphamide administration & dosage, Cytarabine administration & dosage, Dacarbazine administration & dosage, Dexamethasone administration & dosage, Diagnosis, Differential, Disease-Free Survival, Doxorubicin administration & dosage, Etoposide administration & dosage, False Negative Reactions, False Positive Reactions, Fibrosis, Follow-Up Studies, Hematopoietic Stem Cell Transplantation, Hodgkin Disease mortality, Hodgkin Disease pathology, Hodgkin Disease therapy, Humans, Life Tables, Mediastinal Neoplasms pathology, Mediastinal Neoplasms therapy, Melphalan administration & dosage, Neoplasm Recurrence, Local diagnosis, Predictive Value of Tests, Prednisone administration & dosage, Procarbazine administration & dosage, Prospective Studies, Radiotherapy, Adjuvant, Remission Induction, Salvage Therapy, Transplantation, Autologous, Vinblastine administration & dosage, Vincristine administration & dosage, Fluorodeoxyglucose F18, Hodgkin Disease diagnostic imaging, Mediastinal Neoplasms diagnostic imaging, Radiopharmaceuticals, Tomography, Emission-Computed
- Abstract
Residual mediastinal masses are frequently observed in patients with Hodgkin disease (HD) after completed therapy, and the discrimination between active tumor tissue and fibrotic residues remains a clinical challenge. We studied the diagnostic value of metabolic imaging by 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) in detecting active mediastinal disease and predicting relapse. Twenty-eight HD patients with a residual mediastinal mass of at least 2 cm after initial therapy or after salvage chemotherapy were prospectively assigned to 29 examinations with FDG PET and were evaluated as 29 "subjects." Patients were monitored for at least 1 year after examination and observed for signs of relapse. Median follow-up was 28 months (range, 16 to 68 months). A PET-negative mediastinal tumor was observed in 19 subjects, of whom 16 stayed in remission and 3 relapsed. Progression or relapse occurred in 6 of 10 subjects with a positive PET, whereas 4 subjects remained in remission. The negative predictive value (negative PET result and remission) at 1 year was 95%, and the positive predictive value (positive PET result and relapse) was 60%. The disease-free survival for PET-negative and PET-positive patients at 1 year was 95% and 40%, respectively. The difference was statistically significant. A negative FDG PET indicates that an HD patient with a residual mediastinal mass is unlikely to relapse before 1 year, if ever. On the other hand, a positive PET result indicates a significantly higher risk of relapse and demands further diagnostic procedures and a closer follow-up.
- Published
- 2001
- Full Text
- View/download PDF
46. [German Hodgkin's Lymphoma Study Group].
- Author
-
Ansén S, Engert A, Wolf J, Sieber M, Paulus U, and Diehl V
- Subjects
- Combined Modality Therapy, Disease-Free Survival, Forecasting, Germany, Hodgkin Disease diagnosis, Hodgkin Disease mortality, Hodgkin Disease pathology, Humans, Neoplasm Staging, Pilot Projects, Prospective Studies, Hodgkin Disease therapy, Randomized Controlled Trials as Topic trends
- Published
- 2001
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.