T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells
Background Patients with cancers that exhibit extraordinarily high somatic mutation numbers are ideal candidates for immunotherapy and enable identifying tumor-specific peptides through stimulation of tumor-reactive T cells (Tc).Methods Colorectal cancers (CRC) HROC113 and HROC285 were selected base...
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMJ Publishing Group
2022-12-01
|
| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/10/12/e005651.full |
| _version_ | 1797987299759226880 |
|---|---|
| author | Peter Bauer Hans-Georg Rammensee Nadine Mockel-Tenbrinck Andrzej Dzionek Andreas Schlosser Michael Ghosh Markus W Löffler Sandra Schwarz Johanna Schmitz Evgenia Olshvang Marvin Markel Susann Krake Basak Arslan Kapil Dev Kampe Anne Wendt Christina S Mullins Michael Linnebacher |
| author_facet | Peter Bauer Hans-Georg Rammensee Nadine Mockel-Tenbrinck Andrzej Dzionek Andreas Schlosser Michael Ghosh Markus W Löffler Sandra Schwarz Johanna Schmitz Evgenia Olshvang Marvin Markel Susann Krake Basak Arslan Kapil Dev Kampe Anne Wendt Christina S Mullins Michael Linnebacher |
| author_sort | Peter Bauer |
| collection | DOAJ |
| description | Background Patients with cancers that exhibit extraordinarily high somatic mutation numbers are ideal candidates for immunotherapy and enable identifying tumor-specific peptides through stimulation of tumor-reactive T cells (Tc).Methods Colorectal cancers (CRC) HROC113 and HROC285 were selected based on high TMB, microsatellite instability and HLA class I expression. Their HLA ligandome was characterized using mass spectrometry, compared with the HLA ligand atlas and HLA class I-binding affinity was predicted. Cryptic peptides were identified using Peptide-PRISM. Patients’ Tc were isolated from either peripheral blood (pTc) or tumor material (tumor-infiltrating Tc, TiTc) and expanded. In addition, B-lymphoblastoid cells (B-LCL) were generated and used as antigen-presenting cells. pTc and TiTc were stimulated twice for 7 days using peptide pool-loaded B-LCL. Subsequently, interferon gamma (IFNγ) release was quantified by ELISpot. Finally, cytotoxicity against autologous tumor cells was assessed in a degranulation assay.Results 100 tumor-specific candidate peptides—97 cryptic peptides and 3 classically mutated neoantigens—were selected. The neoantigens originated from single nucleotide substitutions in the genes IQGAP1, CTNNB1, and TRIT1. Cryptic and neoantigenic peptides inducing IFNγ secretion of Tc were further investigated. Stimulation of pTc and TiTc with neoantigens and selected cryptic peptides resulted in increased release of cytotoxic granules in the presence of autologous tumor cells, substantiating their improved tumor cell recognition. Tetramer staining showed an enhanced number of pTc and TiTc specific for the IQGAP1 neoantigen. Subpopulation analysis prior to peptide stimulation revealed that pTc mainly consisted of memory Tc, whereas TiTc constituted primarily of effector and effector memory Tc. This allows to infer that TiTc reacting to neoantigens and cryptic peptides must be present within the tumor microenvironment.Conclusion These results prove that the analyzed CRC present both mutated neoantigenic and cryptic peptides on their HLA class I molecules. Moreover, stimulation with these peptides significantly strengthened tumor cell recognition by Tc. Since the overall number of neoantigenic peptides identifiable by HLA ligandome analysis hitherto is small, our data emphasize the relevance of increasing the target scope for cancer vaccines by the cryptic peptide category. |
| first_indexed | 2024-04-11T07:46:43Z |
| format | Article |
| id | doaj.art-e440eaa0b4e347af8ad56649eb46ec22 |
| institution | Directory Open Access Journal |
| issn | 2051-1426 |
| language | English |
| last_indexed | 2024-04-11T07:46:43Z |
| publishDate | 2022-12-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj.art-e440eaa0b4e347af8ad56649eb46ec222022-12-22T04:36:17ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-12-01101210.1136/jitc-2022-005651T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cellsPeter Bauer0Hans-Georg Rammensee1Nadine Mockel-Tenbrinck2Andrzej Dzionek3Andreas Schlosser4Michael Ghosh5Markus W Löffler6Sandra Schwarz7Johanna Schmitz8Evgenia Olshvang9Marvin Markel10Susann Krake11Basak Arslan12Kapil Dev Kampe13Anne Wendt14Christina S Mullins15Michael Linnebacher16Medical Reporting & Genomic Research, Centogene GmbH, Rostock, GermanyInstitute for Cell Biology, Department of Immunology, University of Tübingen, Tubingen, GermanyAff1 grid.59409.310000000405525033Miltenyi Biotec Bergisch Gladbach GermanyAff6 0000 0004 0552 5033grid.59409.31Miltenyi Biotec GmbH Bergisch-Gladbach GermanyRudolf-Virchow Zentrum, University Würzburg, Würzburg, Germany5University of Tübingen, Interfaculty Institute for Cell Biology, Tübingen, GermanyDepartment of General, Visceral and Transplant Surgery, University Hospital, Tübingen, GermanyDepartment of General Surgery, Molecular Oncology and Immunotherapy, Rostock University Medical Center, Rostock, GermanyDepartment of General Surgery, Molecular Oncology and Immunotherapy, Rostock University Medical Center, Rostock, GermanyMiltenyi Biotec BV & Co KG, Bergisch Gladbach, GermanyMiltenyi Biotec BV & Co KG, Bergisch Gladbach, GermanyCentogene GmbH, Rostock, GermanyCentogene GmbH, Rostock, GermanyCentogene GmbH, Rostock, GermanyCentogene GmbH, Rostock, GermanyDepartment of General Surgery, Molecular Oncology and Immunotherapy, Rostock University Medical Center, Rostock, GermanyDepartment of General Surgery, Molecular Oncology and Immunotherapy, Rostock University Medical Center, Rostock, GermanyBackground Patients with cancers that exhibit extraordinarily high somatic mutation numbers are ideal candidates for immunotherapy and enable identifying tumor-specific peptides through stimulation of tumor-reactive T cells (Tc).Methods Colorectal cancers (CRC) HROC113 and HROC285 were selected based on high TMB, microsatellite instability and HLA class I expression. Their HLA ligandome was characterized using mass spectrometry, compared with the HLA ligand atlas and HLA class I-binding affinity was predicted. Cryptic peptides were identified using Peptide-PRISM. Patients’ Tc were isolated from either peripheral blood (pTc) or tumor material (tumor-infiltrating Tc, TiTc) and expanded. In addition, B-lymphoblastoid cells (B-LCL) were generated and used as antigen-presenting cells. pTc and TiTc were stimulated twice for 7 days using peptide pool-loaded B-LCL. Subsequently, interferon gamma (IFNγ) release was quantified by ELISpot. Finally, cytotoxicity against autologous tumor cells was assessed in a degranulation assay.Results 100 tumor-specific candidate peptides—97 cryptic peptides and 3 classically mutated neoantigens—were selected. The neoantigens originated from single nucleotide substitutions in the genes IQGAP1, CTNNB1, and TRIT1. Cryptic and neoantigenic peptides inducing IFNγ secretion of Tc were further investigated. Stimulation of pTc and TiTc with neoantigens and selected cryptic peptides resulted in increased release of cytotoxic granules in the presence of autologous tumor cells, substantiating their improved tumor cell recognition. Tetramer staining showed an enhanced number of pTc and TiTc specific for the IQGAP1 neoantigen. Subpopulation analysis prior to peptide stimulation revealed that pTc mainly consisted of memory Tc, whereas TiTc constituted primarily of effector and effector memory Tc. This allows to infer that TiTc reacting to neoantigens and cryptic peptides must be present within the tumor microenvironment.Conclusion These results prove that the analyzed CRC present both mutated neoantigenic and cryptic peptides on their HLA class I molecules. Moreover, stimulation with these peptides significantly strengthened tumor cell recognition by Tc. Since the overall number of neoantigenic peptides identifiable by HLA ligandome analysis hitherto is small, our data emphasize the relevance of increasing the target scope for cancer vaccines by the cryptic peptide category.https://jitc.bmj.com/content/10/12/e005651.full |
| spellingShingle | Peter Bauer Hans-Georg Rammensee Nadine Mockel-Tenbrinck Andrzej Dzionek Andreas Schlosser Michael Ghosh Markus W Löffler Sandra Schwarz Johanna Schmitz Evgenia Olshvang Marvin Markel Susann Krake Basak Arslan Kapil Dev Kampe Anne Wendt Christina S Mullins Michael Linnebacher T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells Journal for ImmunoTherapy of Cancer |
| title | T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| title_full | T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| title_fullStr | T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| title_full_unstemmed | T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| title_short | T cells of colorectal cancer patients’ stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| title_sort | t cells of colorectal cancer patients stimulated by neoantigenic and cryptic peptides better recognize autologous tumor cells |
| url | https://jitc.bmj.com/content/10/12/e005651.full |
| work_keys_str_mv | AT peterbauer tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT hansgeorgrammensee tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT nadinemockeltenbrinck tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT andrzejdzionek tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT andreasschlosser tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT michaelghosh tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT markuswloffler tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT sandraschwarz tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT johannaschmitz tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT evgeniaolshvang tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT marvinmarkel tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT susannkrake tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT basakarslan tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT kapildevkampe tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT annewendt tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT christinasmullins tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells AT michaellinnebacher tcellsofcolorectalcancerpatientsstimulatedbyneoantigenicandcrypticpeptidesbetterrecognizeautologoustumorcells |