Identification of T cell target antigens in glioblastoma stem-like cells using an integrated proteomics-based approach in patient specimens.
Glioblastoma (GBM) is a highly aggressive brain tumor and still remains incurable. Among others, an immature subpopulation of self-renewing and therapy-resistant tumor cells-often referred to as glioblastoma stem-like cells (GSCs)-has been shown to contribute to disease recurrence. To target these cells personalized immunotherapy has gained a lot of interest, e.g. by reactivating pre-existing anti-tumor immune responses against GSC antigens. To identify T cell targets commonly presented by GSCs and their differentiated counterpart, we used a proteomics-based separation of GSC proteins in combination with a T cell activation assay. Altogether, 713 proteins were identified by LC-ESI-MS/MS mass spectrometry. After a thorough filtering process, 32 proteins were chosen for further analyses. Immunogenicity of corresponding peptides was tested ex vivo. A considerable number of these antigens induced T cell responses in GBM patients but not in healthy donors. Moreover, most of them were overexpressed in primary GBM and also highly expressed in recurrent GBM tissues. Interestingly, expression of the most frequent T cell target antigens could also be confirmed in quiescent, slow-cycling GSCs isolated in high purity by the DEPArray technology. Finally, for a subset of these T cell target antigens, an association between expression levels and higher T cell infiltration as well as an increased expression of positive immune modulators was observed. In summary, we identified novel immunogenic proteins, which frequently induce tumor-specific T cell responses in GBM patients and were also detected in vitro in therapy-resistant quiescent, slow-cycling GSCs. Stable expression of these T cell targets in primary and recurrent GBM support their suitability for future clinical use.
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Carmen
C
Rapp
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Rolf
R
Warta
Translational Immunology Department, German Cancer Research Center, Heidelberg, Germany.
Slava
S
Stamova
German Cancer Consortium (DKTK), Heidelberg, Germany.|Molecular and Translational Radiation Oncology, National Center for Tumor Diseases (NCT), Heidelberg University Hospital and German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Radiation Oncology, Heidelberg University Medical School, Heidelberg Institute of Radiation Oncology (HIRO), National Center for Radiation Research in Oncology (NCOR), Heidelberg, Germany.
Ali
A
Nowrouzi
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Christoph
C
Geisenberger
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Zoltan
Z
Gal
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Saskia
S
Roesch
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Steffen
S
Dettling
Translational Immunology Department, German Cancer Research Center, Heidelberg, Germany.
Simone
S
Juenger
Translational Immunology Department, German Cancer Research Center, Heidelberg, Germany.
Mariana
M
Bucur
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.|German Cancer Consortium (DKTK), Heidelberg, Germany.
Christine
C
Jungk
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Philip
P
DaoTrong
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Rezvan
R
Ahmadi
German Cancer Consortium (DKTK), Heidelberg, Germany.|Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Neuropathology, Institute of Pathology, Ruprecht-Karls-University, Heidelberg, Germany.
Felix
F
Sahm
German Cancer Consortium (DKTK), Heidelberg, Germany.|Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Neuropathology, Institute of Pathology, Ruprecht-Karls-University, Heidelberg, Germany.
David
D
Reuss
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Valentina
V
Fermi
Institute of Pathology, Ruprecht-Karls-University, Heidelberg, Germany.|NCT Tissue Bank, National Center for Tumor Diseases (NCT), Heidelberg, Germany.
Esther
E
Herpel
Department of Internal Medicine V, Heidelberg University Hospital, Heidelberg, Germany.
Volker
V
Eckstein
Hamamatsu Tissue Imaging and Analysis Center (TIGA), BIOQUANT, University of Heidelberg, Heidelberg, Germany.
Niels
N
Grabe
Department of Anaesthesiology, Heidelberg University Hospital, Heidelberg, Germany.
Christoph
C
Schramm
Department of Anaesthesiology, Heidelberg University Hospital, Heidelberg, Germany.
Markus A
MA
Weigand
German Cancer Consortium (DKTK), Heidelberg, Germany.|Molecular and Translational Radiation Oncology, National Center for Tumor Diseases (NCT), Heidelberg University Hospital and German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Radiation Oncology, Heidelberg University Medical School, Heidelberg Institute of Radiation Oncology (HIRO), National Center for Radiation Research in Oncology (NCOR), Heidelberg, Germany.
Juergen
J
Debus
German Cancer Consortium (DKTK), Heidelberg, Germany.|Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Neuropathology, Institute of Pathology, Ruprecht-Karls-University, Heidelberg, Germany.
Andreas
A
von Deimling
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany.
Andreas
A
Unterberg
German Cancer Consortium (DKTK), Heidelberg, Germany.|Molecular and Translational Radiation Oncology, National Center for Tumor Diseases (NCT), Heidelberg University Hospital and German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Radiation Oncology, Heidelberg University Medical School, Heidelberg Institute of Radiation Oncology (HIRO), National Center for Radiation Research in Oncology (NCOR), Heidelberg, Germany.
Amir
A
Abdollahi
Translational Immunology Department, German Cancer Research Center, Heidelberg, Germany.
Philipp
P
Beckhove
Division of Experimental Neurosurgery, Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany. H.Mende@med.uni-heidelberg.de.|German Cancer Consortium (DKTK), Heidelberg, Germany. H.Mende@med.uni-heidelberg.de.|Sektion Neurochirurgische Forschung, Neurochirurgische Universitätsklinik, INF400, 69120, Heidelberg, Germany. H.Mende@med.uni-heidelberg.de.
Christel
C
Herold-Mende
D000818:Animals; D017305:Annexin A1 / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001932:Brain Neoplasms / Q000473:pathology*; D015197:Carcinogenicity Tests; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000378:metabolism; D055312:Cystatin A / Q000378:metabolism; D004195:Disease Models, Animal; D018604:Epitope Mapping; D005260:Female; D005909:Glioblastoma / Q000473:pathology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000473:pathology; D040901:Proteomics; D016176:T-Lymphocyte Subsets / Q000473:pathology*
D017305:Annexin A1; D000951:Antigens, Neoplasm; D002352:Carrier Proteins; D018834:Chaperonin 60; D055312:Cystatin A; C502658:FSCN1 protein, human; C060293:HSPD1 protein, human; D019394:Ki-67 Antigen; D008840:Microfilament Proteins; D024101:Mitochondrial Proteins; C026407:CSTA protein, human; D007371:Interferon-gamma
Heterogeneity; IDH1-wt glioblastoma; Plasticity; Quiescent stem-like cells; T cell target antigen repertoire