- Email: [email protected]
Biomarkers of immune switch induced by a novel anti-macrophage antibody (anti-Clever-1 mAb; FP-1305) in MATINS trial patients with advanced solid tumours
Annals of Oncology 30 (Supplement 11): xi1–xi11, 2019 doi:10.1093/annonc/mdz447
1O
Harmonization and standardization of panel-based tumour mutational burden (TMB) measurement: Real-world results and recommendations of the QuIP study
A. Stenzinger1, V. Endris1, J. Budczies1, S. Merkelbach-Bruse2, W. Dietmaier3, U. Siebolts4, J. Maas5, D.M. Merino6, M. Stewart6, J. Allen6, H. Glimm7, M. Thiemann8, D. Aust9, M. Hummel10, H. Moch11, A. Jung12, F. Haller13, W. Weichert12, M. Dietel10 1 Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany, 2Institute of Pathology, University Hospital Cologne, Cologne, Germany, 3Institute of Pathology, University Hospital Regensburg, Regensburg, Germany, 4Institute of Pathology, University Hospital Halle, Halle, Germany, 5Quality in Pathology GmbH, Berlin, Germany, 6Friends of Cancer Research (FoCR), Washington, DC, USA, 7National Center for Tumor Diseases Dresden, University Hospital Dresden, Dresden, Germany, 8Institute of Pathology, Hematopathology Hamburg, Hamburg, Germany, 9Institute of Pathology, University Hospital Dresden, Dresden, Germany, 10Institute of Pathology, Charite´ University Hospital, Berlin, Germany, 11Department of Pathology and Molecular Pathology, University Hospital Zurich, Zurich, Switzerland, 12Institute of Pathology, University Hospital Munich (LMU), Munich, Germany, 13Institute of Pathology, University Hospital Erlangen, Erlangen, Germany Background: TMB is a novel predictive biomarker that can identify patients who may benefit from immunotherapy. NSCLC trial data suggest that whole exome sequencing (WES) and panel-sequencing are suitable to determine TMB, and that centralized and decentralized/lab-developed testing models for panel-based measurements are acceptable. In strategic partnership with the effort led by Friends of Cancer Research, the Quality in Pathology (QuIP) study was designed to analyze performance and specifications of TMB panels in a wet-lab setting. Methods: 20 FFPE samples (NSCLC, HNSCC, CRC, including MSI, mutant POLE) that cover the full spectrum of TMB (2 to 200 muts/Mb) were analyzed by 11 pathology centers and 4 assay providers using 6 different major panels. WES data served as reference standard. Each tumor sample was tested > 20 times across several panels and institutions resulting in 580 datasets. Using raw sequencing data, processed file formats, and reported TMB values, we dissected specifications of each panel result, identified panel-specific requirements, and analyzed Pearson correlations of TMB data between assays, labs, and vs WES. Results: Each panel had different requirements regarding library preparation (hybridization vs PCR) and input material (range: 20-200 ng). We identified tumor cell content and DNA quality/quantity as crucial preanalytic factors that require integration with coverage data, VAF cut-points, and assay-specific features (eg, molecular barcodes) to obtain reliable TMB results. Control of C>T artifacts was important for assays not using molecular identifiers. Correlations between panel-TMB estimates (R ¼ 0.93 6 0.1; mean 6 sd) and with WES (R > 0.9 for 18 and R > 0.95 for 14 of 20 panel tests) were strong and improved after optimization of pipelines. Conclusion: The QuIP study demonstrated that all TMB panels work under real-world conditions and strongly correlate with WES data, with low variability across sites. Further, we identified both common and panel-specific parameters that influence TMB results in daily practice. Recommendations will be provided that support standardization and enable implementation of TMB testing in routine diagnostics. Legal entity responsible for the study: The authors. Funding: Bristol-Myers Squibb, Roche, Illumina, Thermo Fisher, Neo Oncology, Qiagen. Disclosure: A. Stenzinger: Advisory / Consultancy, Speaker Bureau / Expert testimony: Bristol-Myers Squibb; Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca; Advisory / Consultancy, Speaker Bureau / Expert testimony: Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Novartis; Advisory / Consultancy, Speaker Bureau / Expert testimony: Illumina; Advisory / Consultancy, Speaker Bureau / Expert testimony: Thermo Fisher; Speaker Bureau / Expert testimony: Roche; Speaker Bureau / Expert testimony: Pfizer; Advisory / Consultancy: Seattle Genomics; Speaker Bureau / Expert testimony: MSD. All other authors have declared no conflicts of interest.
2O
Biomarkers of immune switch induced by a novel anti-macrophage antibody (anti-Clever-1 mAb; FP-1305) in MATINS trial patients with advanced solid tumours
M. Hollme´n1, R. Virtakoivu1, P. Jaakkola2, A. Minchom3, S. Jalkanen1, M. Karvonen4, J. Mandelin5, J. Koivunen6, P. Bono7 1 Medicity Research Laboratory, University of Turku, Turku, Finland, 2Oncology, Helsinki University Hospital, Helsinki, Finland, 3Drug Development Unit, Royal Marsden Hospital, London, UK, 4Clinical Development, Faron Pharmaceuticals, Turku, Finland, 5Pre-clinical Development, Faron Pharmaceuticals, Turku, Finland, 6Department of Oncology and Radiotherapy, Oulu University Hospital, Oulu, Finland, 7Terveystalo, Helsinki, Finland Background: A scavenger receptor CLEVER-1 is highly expressed on tumor associated macrophages (TAMs) and mediates the clearance of “unwanted” self-components. Pre-clinical studies demonstrate that CLEVER-1 inhibition increases TAM pro-
inflammatory cytokine secretion and antigen presentation reactivating CD8þ T cell responses with robust anti-tumor activity (Viitala et al., 2019). Targeting CLEVER-1 could overcome the immunosuppressive tumor microenvironment and has led to the development of FP-1305, a humanized anti-CLEVER-1 IgG4-antibody. Methods: MATINS (Macrophage Antibody To INhibit immune Suppression) trial is a multicenter first-in-human phase I/II study (NCT03733990) to assess the tolerability, safety and preliminary efficacy of FP-1305 in patients with advanced, IO-refractory melanoma, cholangiocarcinoma, hepatocellular, colorectal, and pancreatic ductal adenocarcinoma. Biomarker analysis included CLEVER-1 determination, immune cell profiling by mass cytometry and analysis of cytokine production. Results: 11 patients (median age 57) were enrolled in four cohorts (0.3, 1.0, 3.0 or 10 mg/kg) and received 1-8 cycles (median 3) of FP-1305 every three weeks. FP-1305 has been well tolerated without dose-limiting toxicities and maximum tolerated dose (MTD) has not been reached. Promising early efficacy results have recently been reported (ESMO 2019, LBA19). FP-1305 dosing led to increased Th1 skewing (CXCR3þCCR6-) of CD4 and CD8 T cell populations with downregulation of several inhibitory immune checkpoint molecules. Increase in circulating IFN gamma was detected but it was most prominent in the patient showing durable partial response. Conclusion: FP-1305 is the first macrophage checkpoint inhibitor candidate promoting immune switch with promising tolerability and clinical anti-tumor activity. FP1305 represents a novel treatment option to provoke immune response against cold tumors. Clinical trial identification: NCT03733990. Legal entity responsible for the study: Faron Pharmaceuticals. Funding: Finnish Academy, Finnish Cancer Foundations, Sigrid Juselius Foundation, Faron Pharmaceuticals. Disclosure: M. Hollme´n: Research grant / Funding (institution), Travel / Accommodation / Expenses, Shareholder / Stockholder / Stock options: Faron Pharmaceuticals. P. Jaakkola: Advisory / Consultancy: Faron Pharmaceuticals. A. Minchom: Advisory / Consultancy: Faron Pharmaceuticals. S. Jalkanen: Shareholder / Stockholder / Stock options: Faron Pharmaceuticals. M. Karvonen: Shareholder / Stockholder / Stock options, Full / Part-time employment: Faron Pharmaceuticals. J. Mandelin: Shareholder / Stockholder / Stock options, Full / Part-time employment: Faron Pharmaceuticals. J. Koivunen: Advisory / Consultancy: Faron Pharmaceuticals; Advisory / Consultancy: Novartis; Advisory / Consultancy: Pfizer; Advisory / Consultancy: BoehringerIngelheim; Advisory / Consultancy: KaikuHealth. P. Bono: Advisory / Consultancy, Travel / Accommodation / Expenses, Spouse / Financial dependant: Faron Pharmaceuticals; Advisory / Consultancy, Travel / Accommodation / Expenses: MSD; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Novartis; Advisory / Consultancy: Bristol-Myers Squibb; Advisory / Consultancy: OrionPharma. All other authors have declared no conflicts of interest.
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A pre-existing inflammatory immune microenvironment predicts the clinical and immunological response of vulvar high-grade squamous intraepithelial lesions to therapeutic HPV16 peptide vaccination
Z. Abdulrahman1, N.F.C.C. de Miranda2, M.I.E. van Poelgeest3, S.H. van der Burg4 Medical Oncology, Gynaecology and Pathology, Leiden University Medical Center, Leiden, Netherlands, 2Pathology, Leiden University Medical Center, Leiden, Netherlands, 3 Gynaecology, Leiden University Medical Center, Leiden, Netherlands, 4Medical Oncology, Leiden University Medical Center, Leiden, Netherlands
1
Background: Vulvar High-grade Squamous Intraepithelial Lesion (vHSIL) is predominantly induced by high-risk Human Papilloma Virus type 16 (HPV16). In two independent trials, therapeutic vaccination against the oncoproteins of HPV16 with synthetic long peptides (SLP) resulted in vHSIL regression in about half of the patients after 12 months. Several studies have shown that the immune microenvironment influences therapy outcome. Therefore, a thorough investigation of the vHSIL immune microenvironment before and after SLP vaccination was performed, and its impact on clinical response was studied. Methods: Two novel multiplex immunofluorescence panels were designed for formalin-fixed paraffin-embedded tissue, one for T cells (CD3, CD8, FoxP3, Tim3, Tbet, PD-1, DAPI) and one for myeloid cells (CD14, CD33, CD68, CD163, CD11c, PD-L1, DAPI). Pre- and 3 months post-vaccination biopsies of 29 patients and 27 healthy vulva excisions were stained, scanned with the Vectra multispectral imaging system, and automatically phenotyped and counted with inForm advanced image analysis software. Results: A pre-existing pro-inflammatory TME, marked by high numbers of CD4 and CD8 T cells expressing Tbet and/or PD-1 as well as CD14þ inflammatory macrophages, is a strong predictor for good clinical response. A clear stepwise increase in prevaccination infiltrating Tbetþ, CD4þ, CD8þ T cells and CD14þ macrophages, and decrease in Foxp3þ Tregs was observed as response increased from non to partial to complete response. Moreover, the pre-vaccination immune microenvironment of complete responders resembled healthy vulva. Vaccination further increased
C European Society for Medical Oncology 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology. V
All rights reserved. For permissions, please email: [email protected].
Downloaded from https://academic.oup.com/annonc/article-abstract/30/Supplement_11/mdz447.001/5675155 by University of Saskatchewan user on 16 December 2019
BIOMARKER DEVELOPMENT
1O
Harmonization and standardization of panel-based tumour mutational burden (TMB) measurement: Real-world results and recommendations of the QuIP study
A. Stenzinger1, V. Endris1, J. Budczies1, S. Merkelbach-Bruse2, W. Dietmaier3, U. Siebolts4, J. Maas5, D.M. Merino6, M. Stewart6, J. Allen6, H. Glimm7, M. Thiemann8, D. Aust9, M. Hummel10, H. Moch11, A. Jung12, F. Haller13, W. Weichert12, M. Dietel10 1 Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany, 2Institute of Pathology, University Hospital Cologne, Cologne, Germany, 3Institute of Pathology, University Hospital Regensburg, Regensburg, Germany, 4Institute of Pathology, University Hospital Halle, Halle, Germany, 5Quality in Pathology GmbH, Berlin, Germany, 6Friends of Cancer Research (FoCR), Washington, DC, USA, 7National Center for Tumor Diseases Dresden, University Hospital Dresden, Dresden, Germany, 8Institute of Pathology, Hematopathology Hamburg, Hamburg, Germany, 9Institute of Pathology, University Hospital Dresden, Dresden, Germany, 10Institute of Pathology, Charite´ University Hospital, Berlin, Germany, 11Department of Pathology and Molecular Pathology, University Hospital Zurich, Zurich, Switzerland, 12Institute of Pathology, University Hospital Munich (LMU), Munich, Germany, 13Institute of Pathology, University Hospital Erlangen, Erlangen, Germany Background: TMB is a novel predictive biomarker that can identify patients who may benefit from immunotherapy. NSCLC trial data suggest that whole exome sequencing (WES) and panel-sequencing are suitable to determine TMB, and that centralized and decentralized/lab-developed testing models for panel-based measurements are acceptable. In strategic partnership with the effort led by Friends of Cancer Research, the Quality in Pathology (QuIP) study was designed to analyze performance and specifications of TMB panels in a wet-lab setting. Methods: 20 FFPE samples (NSCLC, HNSCC, CRC, including MSI, mutant POLE) that cover the full spectrum of TMB (2 to 200 muts/Mb) were analyzed by 11 pathology centers and 4 assay providers using 6 different major panels. WES data served as reference standard. Each tumor sample was tested > 20 times across several panels and institutions resulting in 580 datasets. Using raw sequencing data, processed file formats, and reported TMB values, we dissected specifications of each panel result, identified panel-specific requirements, and analyzed Pearson correlations of TMB data between assays, labs, and vs WES. Results: Each panel had different requirements regarding library preparation (hybridization vs PCR) and input material (range: 20-200 ng). We identified tumor cell content and DNA quality/quantity as crucial preanalytic factors that require integration with coverage data, VAF cut-points, and assay-specific features (eg, molecular barcodes) to obtain reliable TMB results. Control of C>T artifacts was important for assays not using molecular identifiers. Correlations between panel-TMB estimates (R ¼ 0.93 6 0.1; mean 6 sd) and with WES (R > 0.9 for 18 and R > 0.95 for 14 of 20 panel tests) were strong and improved after optimization of pipelines. Conclusion: The QuIP study demonstrated that all TMB panels work under real-world conditions and strongly correlate with WES data, with low variability across sites. Further, we identified both common and panel-specific parameters that influence TMB results in daily practice. Recommendations will be provided that support standardization and enable implementation of TMB testing in routine diagnostics. Legal entity responsible for the study: The authors. Funding: Bristol-Myers Squibb, Roche, Illumina, Thermo Fisher, Neo Oncology, Qiagen. Disclosure: A. Stenzinger: Advisory / Consultancy, Speaker Bureau / Expert testimony: Bristol-Myers Squibb; Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca; Advisory / Consultancy, Speaker Bureau / Expert testimony: Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Novartis; Advisory / Consultancy, Speaker Bureau / Expert testimony: Illumina; Advisory / Consultancy, Speaker Bureau / Expert testimony: Thermo Fisher; Speaker Bureau / Expert testimony: Roche; Speaker Bureau / Expert testimony: Pfizer; Advisory / Consultancy: Seattle Genomics; Speaker Bureau / Expert testimony: MSD. All other authors have declared no conflicts of interest.
2O
Biomarkers of immune switch induced by a novel anti-macrophage antibody (anti-Clever-1 mAb; FP-1305) in MATINS trial patients with advanced solid tumours
M. Hollme´n1, R. Virtakoivu1, P. Jaakkola2, A. Minchom3, S. Jalkanen1, M. Karvonen4, J. Mandelin5, J. Koivunen6, P. Bono7 1 Medicity Research Laboratory, University of Turku, Turku, Finland, 2Oncology, Helsinki University Hospital, Helsinki, Finland, 3Drug Development Unit, Royal Marsden Hospital, London, UK, 4Clinical Development, Faron Pharmaceuticals, Turku, Finland, 5Pre-clinical Development, Faron Pharmaceuticals, Turku, Finland, 6Department of Oncology and Radiotherapy, Oulu University Hospital, Oulu, Finland, 7Terveystalo, Helsinki, Finland Background: A scavenger receptor CLEVER-1 is highly expressed on tumor associated macrophages (TAMs) and mediates the clearance of “unwanted” self-components. Pre-clinical studies demonstrate that CLEVER-1 inhibition increases TAM pro-
inflammatory cytokine secretion and antigen presentation reactivating CD8þ T cell responses with robust anti-tumor activity (Viitala et al., 2019). Targeting CLEVER-1 could overcome the immunosuppressive tumor microenvironment and has led to the development of FP-1305, a humanized anti-CLEVER-1 IgG4-antibody. Methods: MATINS (Macrophage Antibody To INhibit immune Suppression) trial is a multicenter first-in-human phase I/II study (NCT03733990) to assess the tolerability, safety and preliminary efficacy of FP-1305 in patients with advanced, IO-refractory melanoma, cholangiocarcinoma, hepatocellular, colorectal, and pancreatic ductal adenocarcinoma. Biomarker analysis included CLEVER-1 determination, immune cell profiling by mass cytometry and analysis of cytokine production. Results: 11 patients (median age 57) were enrolled in four cohorts (0.3, 1.0, 3.0 or 10 mg/kg) and received 1-8 cycles (median 3) of FP-1305 every three weeks. FP-1305 has been well tolerated without dose-limiting toxicities and maximum tolerated dose (MTD) has not been reached. Promising early efficacy results have recently been reported (ESMO 2019, LBA19). FP-1305 dosing led to increased Th1 skewing (CXCR3þCCR6-) of CD4 and CD8 T cell populations with downregulation of several inhibitory immune checkpoint molecules. Increase in circulating IFN gamma was detected but it was most prominent in the patient showing durable partial response. Conclusion: FP-1305 is the first macrophage checkpoint inhibitor candidate promoting immune switch with promising tolerability and clinical anti-tumor activity. FP1305 represents a novel treatment option to provoke immune response against cold tumors. Clinical trial identification: NCT03733990. Legal entity responsible for the study: Faron Pharmaceuticals. Funding: Finnish Academy, Finnish Cancer Foundations, Sigrid Juselius Foundation, Faron Pharmaceuticals. Disclosure: M. Hollme´n: Research grant / Funding (institution), Travel / Accommodation / Expenses, Shareholder / Stockholder / Stock options: Faron Pharmaceuticals. P. Jaakkola: Advisory / Consultancy: Faron Pharmaceuticals. A. Minchom: Advisory / Consultancy: Faron Pharmaceuticals. S. Jalkanen: Shareholder / Stockholder / Stock options: Faron Pharmaceuticals. M. Karvonen: Shareholder / Stockholder / Stock options, Full / Part-time employment: Faron Pharmaceuticals. J. Mandelin: Shareholder / Stockholder / Stock options, Full / Part-time employment: Faron Pharmaceuticals. J. Koivunen: Advisory / Consultancy: Faron Pharmaceuticals; Advisory / Consultancy: Novartis; Advisory / Consultancy: Pfizer; Advisory / Consultancy: BoehringerIngelheim; Advisory / Consultancy: KaikuHealth. P. Bono: Advisory / Consultancy, Travel / Accommodation / Expenses, Spouse / Financial dependant: Faron Pharmaceuticals; Advisory / Consultancy, Travel / Accommodation / Expenses: MSD; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Novartis; Advisory / Consultancy: Bristol-Myers Squibb; Advisory / Consultancy: OrionPharma. All other authors have declared no conflicts of interest.
3O
A pre-existing inflammatory immune microenvironment predicts the clinical and immunological response of vulvar high-grade squamous intraepithelial lesions to therapeutic HPV16 peptide vaccination
Z. Abdulrahman1, N.F.C.C. de Miranda2, M.I.E. van Poelgeest3, S.H. van der Burg4 Medical Oncology, Gynaecology and Pathology, Leiden University Medical Center, Leiden, Netherlands, 2Pathology, Leiden University Medical Center, Leiden, Netherlands, 3 Gynaecology, Leiden University Medical Center, Leiden, Netherlands, 4Medical Oncology, Leiden University Medical Center, Leiden, Netherlands
1
Background: Vulvar High-grade Squamous Intraepithelial Lesion (vHSIL) is predominantly induced by high-risk Human Papilloma Virus type 16 (HPV16). In two independent trials, therapeutic vaccination against the oncoproteins of HPV16 with synthetic long peptides (SLP) resulted in vHSIL regression in about half of the patients after 12 months. Several studies have shown that the immune microenvironment influences therapy outcome. Therefore, a thorough investigation of the vHSIL immune microenvironment before and after SLP vaccination was performed, and its impact on clinical response was studied. Methods: Two novel multiplex immunofluorescence panels were designed for formalin-fixed paraffin-embedded tissue, one for T cells (CD3, CD8, FoxP3, Tim3, Tbet, PD-1, DAPI) and one for myeloid cells (CD14, CD33, CD68, CD163, CD11c, PD-L1, DAPI). Pre- and 3 months post-vaccination biopsies of 29 patients and 27 healthy vulva excisions were stained, scanned with the Vectra multispectral imaging system, and automatically phenotyped and counted with inForm advanced image analysis software. Results: A pre-existing pro-inflammatory TME, marked by high numbers of CD4 and CD8 T cells expressing Tbet and/or PD-1 as well as CD14þ inflammatory macrophages, is a strong predictor for good clinical response. A clear stepwise increase in prevaccination infiltrating Tbetþ, CD4þ, CD8þ T cells and CD14þ macrophages, and decrease in Foxp3þ Tregs was observed as response increased from non to partial to complete response. Moreover, the pre-vaccination immune microenvironment of complete responders resembled healthy vulva. Vaccination further increased
C European Society for Medical Oncology 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology. V
All rights reserved. For permissions, please email: [email protected].
Downloaded from https://academic.oup.com/annonc/article-abstract/30/Supplement_11/mdz447.001/5675155 by University of Saskatchewan user on 16 December 2019
BIOMARKER DEVELOPMENT