- Email: [email protected]
Novel dendritic cell based immunotherapy for advanced cancer
abstracts & Dohme; Advisory/Consultancy, Research grant/Funding (self): Array; Advisory/Consultancy: Novartis; Advisory/Consultancy: Amgen; Advisory/Consultancy: Merck Serono; Advisory/ Consultancy: Pierre Fabre; Advisory/Consultancy: Incyte; Advisory/Consultancy: NewLink Genetics; Advisory/Consultancy: Genmab; Advisory/Consultancy: Medimmune; Advisory/Consultancy: AstraZeneca; Advisory/Consultancy: Syndax; Advisory/Consultancy: SunPharma; Advisory/ Consultancy: Sanofi; Advisory/Consultancy: Idera. C. Robert: Advisory/Consultancy: Roche; Advisory/Consultancy: GSK; Advisory/Consultancy: Merck Sharp & Dohme; Advisory/Consultancy: Novartis; Advisory/Consultancy: Amgen; Advisory/Consultancy: BMS. N. Chaput: Research grant/ Funding (self): Cytune Pharma; Research grant/Funding (self): GSK; Speaker Bureau/Expert testimony, Research grant/Funding (self): Sanofi; Advisory/Consultancy, Speaker Bureau/Expert testimony: AstraZeneca. All other authors have declared no conflicts of interest.
Novel dendritic cell based immunotherapy for advanced cancer
A.P. Kaur1, J. Adhikaree2, H. Franks2, P. Patel2, A.M. Jackson3 Academic Oncology, University of Nottingham, School of Medicine, Nottingham, UK, 2 Oncology, Nottingham University Hospitals NHS Trust-City Hospital Campus, Nottingham, UK, 3Academic Oncology, University of Nottingham, Nottingham, UK
1
Background: Immune-checkpoint inhibitors (ICI) such as Ipilumimab and Nivolumab have made long-term survival a real possibility for advanced cancer patients. Despite the success, only a proportion of patients receiving ICI show meaningful clinical benefit. Developing a more personalized treatment strategy using patient’s own immune cells may be a promising approach for treating non-responding patients. Natural circulating dendritic cells (nDC) such as CD1cþ DC (cDC2) subset and plasmacytoid DC (pDC) are essential for efficient activation of immune responses after ICI therapy. Thus understanding the condition of these cells in patients and devising new strategies to improve their functions may help improve response to ICI in advanced cancer patients. Methods: To investigate this we measured the number and phenotype of nDC in 29 advanced cancer patients versus 21 age and gender-matched healthy controls using flow cytometry. We further identified that inhibition of the p38 MAPK pathway using small molecule inhibitors in cDC2 cells increases their immune-stimulatory capacity, measured by functional assays. Results: The number of pDC were significantly reduced in cancer patients as compared to healthy controls (1221 vs 2843 pDC/mL) (p-value<0.01). Although the number of cDC2 were slightly reduced in cancer patients (4476 vs 5053 of cDC2/mL), a good proportion of patients had sufficient number of DC eligible for a DC-based immunotherapy. Patient cDC2 were immune-suppressed with low secretion of immunestimulatory cytokines (IL-12) and impaired homing to lymph-nodes. We identified that the p38 MAPK signalling pathway was controlling immune-suppression in cDC2 cells. Inhibiting this pathway restored the secretion of IL-12 in all patients (n ¼ 5), increased the homing of DC to lymph-node chemokines, and prevented them from secreting the immune-suppressive cytokine, IL-10. Conclusions: The number and phenotype of nDC are suppressed in advanced cancer patients. Inhibition of p38 MAPK can restore the function of patient cDC2 subset. Coculture of these p38-inhibited cDC2 with pDC further show promising results important for developing advanced DC vaccines. Our study has paved the way for a phase I clinical trial of adoptive transfer of p38-MAPK inhibited cDC2 in 24 advanced cancer patients. Legal entity responsible for the study: University of Nottingham. Funding: University of Nottingham. Disclosure: All authors have declared no conflicts of interest.
5PD
The role of EGFR inhibitor (EGFRi) in immune cell infiltration and CD81 T-cell activation in EGFR mutant lung cancer
F. Li1, G. Lizee1, P. Hwu1, X. Du2, L. Deng3, A. Talukder1, A. Katailiha1, Q. Zou3, J. Roszik1, D. Hawke1, K. Jackson1, S. Bradley1, Y. Wang3, R. Ataullakhanov4, A. Bagaev4, N. Kotlov4, V. Svekolkin4, N. Miheecheva4, F. Frenkel4, H. Sonnemann1 1 Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA, 2Oncology Department, Tianjin Beichen Hospital, Tianjin, China, 3Research and Development Department, Tianjin HengJia Biotechnology Development Co., Ltd., Tianjin, China, 4BostonGene Corporation, BostonGene Corporation, Boston, MA, USA Background: Oncogenic MAP kinase pathway activating mutations have been shown to drive immune suppression in melanoma and colorectal cancers. In this study, we explored whether oncogenic EGFR mutations play an analogous role in non-small cell lung cancer (NSCLC), and whether EGFRi can relieve tumor-associated immune suppression. Methods: Lung cancer cell lines expressing wild-type or mutated EGFR were treated with EGFRi, followed by collection of RNA and cell supernatants. In parallel, tumor biopsies from NSCLC patients receiving a personalized peptide vaccine with or without concurrent EGFRi treatment were obtained. RNAseq-based transcriptome profiles of tumor cell lines and patient tumor biopsies were compared to assess common gene signatures driven by EGFRi. Gene expression changes were confirmed at the protein level using Western blot, flow cytometry, and cytokine/chemokine Luminex. The impact of EGFRi on T-cell migration and tumor cell recognition by antigen-specific CD8þ T cells was also assessed.
v2 | Basic Science
Results: In addition to downregulating genes associated with cell proliferation, apoptosis and survival, EGFRi increased the transcription of genes associated with TNFa and TRAIL signaling, and antigen presentation. HLA class I protein upregulation was confirmed and correlated with increased recognition of tumor cells by cytotoxic T cells. Several chemokines and cytokines were up- or down-regulated following EGFRi treatment, and Luminex analysis confirmed changes to 10 of them. Migration assays demonstrated that chemotaxis of T cells towards EGFR-mutant cell supernatants increased in an EGFRi dose-dependent manner. Transcriptome profiling of tumor biopsies revealed similar gene expression changes in on-EGFRi treatment tumor samples. Furthermore, increased tumor immune cell infiltration observed in EGFRi-treated patient tumors was consistent with the upregulation of EGFRi signature chemokines at the tumor site. Conclusions: These results provide evidence that EGFRi has the capacity to facilitate modulation of the tumor microenvironment to favor immune cell infiltration and promote T-cell mediated antitumor immunity. Legal entity responsible for the study: The University of Texas MD Anderson Cancer Center. Funding: Tianjin HengJia Biotechnology Development Co., Ltd. Disclosure: F. Li: Shareholder/Stockholder/Stock options: Tianjin HengJia Biotechnology Development Co., Ltd.. G. Lizee: Advisory/Consultancy: HengJia Neoantigen Biotechnology (Tianjin) Co., Ltd.. P. Hwu: Advisory/Consultancy: Dragonfly Therapeutics; Advisory/Consultancy: GlaxoSmithKline; Advisory/Consultancy, Shareholder/Stockholder/Stock options: Immatics; Advisory / Consultancy: Sanofi; Research grant/Funding (institution): Genentech. L. Deng: Full/Parttime employment: Tianjin HengJia Biotechnology Development Co., Ltd.. Q. Zou: Full/Part-time employment: Tianjin HengJia Biotechnology Development Co., Ltd.. Y. Wang: Full/Part-time employment: Tianjin HengJia Biotechnology Development Co., Ltd.. R. Ataullakhanov: Full/Parttime employment: Bostongene. Llc. Bagaev: Full/Part-time employment: Bostongene. Llc. N. Kotlov: Full/Part-time employment: Bostongene. Llc. V. Svekolkin: Full/Part-time employment: Bostongene. Llc. N. Miheecheva: Full/Part-time employment: Bostongene. Llc. F. Frenkel: Full/Parttime employment: Bostongene. Llc. All other authors have declared no conflicts of interest.
6PD
The mutational signature of spontaneously developing tumours in MLH1-/- mice: Potential consequences for immunotherapeutic approaches
C. Maletzki1, Y. Gladbach2, M. hamed2, C. Junghanss1 Hematology, Oncology, Palliative Medicine, Universit€ atsmedizin Rostock, Rostock, Germany, 2Rostock University Medical Center, Institute for Biostatistics and Informatics in Medicine and Ageing Research (IBIMA), Universit€ atsmedizin Rostock, Rostock, Germany
1
Background: MLH1 knock out mice develop mismatch repair deficient (MMR-D) neoplasias spontaneously and reflect the diverse clinical presentation of MMR-Ddriven carcinogenesis in men. The tumor spectrum includes a high prevalence of early Non-Hodgkin T cell lymphomas (NHL), lymphoid skin lesions as well as later developing epithelial tumors of the gastrointestinal tract (GIT). Methods: Using whole-exome sequencing on MLH1-/- tumors (2x GIT, 1x splenic NHL, 1x skin lymphoma) as well as GIT-derived cell lines (n ¼ 2), we focused on identification of (I) shared and (II) mutually exclusive mutations and described the processes of ongoing mutational events in tumor-derived cultures. Results: MLH1-/- tumors show high tumor mutational burden with 3/4 primary tumor samples even being ultra-hypermutated (> 100 mut/MB). Missense mutations were more frequent than nonsense mutations, base changes were mainly due to transitions (C>T; A>G). The resulting mutational landscape was heterogeneous and in accordance with the human counterpart, MLH1-/- tumors frequently harbor mutations in PIK3CA, EGFR, KRAS, and/or ERBB3. Of note, only a few shared mutations were detectable among different tumor entities, among them were ARID1A and IDH2. Mutations in classical tumor suppressor genes SMAD4 and POLE were mutually exclusive in lymphomas, most likely contributing to a more aggressive in vivo phenotype. Comparing the mutational profile of selected tumors and their corresponding cell line revealed continuous increased numbers of somatic gene mutations. The same was true for coding microsatellite mutations in MMR-D target genes. Partial overlap was detectable, yet recognizing shared antigens. Two promising candidates are AKT3, a RACgamma serine/threonine-protein kinase and the endonuclease ERCC5 (Excision Repair 5), involved in DNA excision repair. Conclusions: The present study is the first reporting results of a comparison between different spontaneously developing tumors as models for MMR-D driven tumorigenesis. Additionally to identifying ARID1A as causative mutation hotspot, this comprehensive characterization of the mutational landscape may be a good starting point to predict antigens for vaccination approaches. Legal entity responsible for the study: The authors. Funding: german research foundation (DFG); grant number: MA5799/2-1. Disclosure: All authors have declared no conflicts of interest.
Volume 30 | Supplement 5 | October 2019
Downloaded from https://academic.oup.com/annonc/article-abstract/30/Supplement_5/mdz238.003/5576228 by guest on 24 October 2019
4O
Annals of Oncology
Novel dendritic cell based immunotherapy for advanced cancer
A.P. Kaur1, J. Adhikaree2, H. Franks2, P. Patel2, A.M. Jackson3 Academic Oncology, University of Nottingham, School of Medicine, Nottingham, UK, 2 Oncology, Nottingham University Hospitals NHS Trust-City Hospital Campus, Nottingham, UK, 3Academic Oncology, University of Nottingham, Nottingham, UK
1
Background: Immune-checkpoint inhibitors (ICI) such as Ipilumimab and Nivolumab have made long-term survival a real possibility for advanced cancer patients. Despite the success, only a proportion of patients receiving ICI show meaningful clinical benefit. Developing a more personalized treatment strategy using patient’s own immune cells may be a promising approach for treating non-responding patients. Natural circulating dendritic cells (nDC) such as CD1cþ DC (cDC2) subset and plasmacytoid DC (pDC) are essential for efficient activation of immune responses after ICI therapy. Thus understanding the condition of these cells in patients and devising new strategies to improve their functions may help improve response to ICI in advanced cancer patients. Methods: To investigate this we measured the number and phenotype of nDC in 29 advanced cancer patients versus 21 age and gender-matched healthy controls using flow cytometry. We further identified that inhibition of the p38 MAPK pathway using small molecule inhibitors in cDC2 cells increases their immune-stimulatory capacity, measured by functional assays. Results: The number of pDC were significantly reduced in cancer patients as compared to healthy controls (1221 vs 2843 pDC/mL) (p-value<0.01). Although the number of cDC2 were slightly reduced in cancer patients (4476 vs 5053 of cDC2/mL), a good proportion of patients had sufficient number of DC eligible for a DC-based immunotherapy. Patient cDC2 were immune-suppressed with low secretion of immunestimulatory cytokines (IL-12) and impaired homing to lymph-nodes. We identified that the p38 MAPK signalling pathway was controlling immune-suppression in cDC2 cells. Inhibiting this pathway restored the secretion of IL-12 in all patients (n ¼ 5), increased the homing of DC to lymph-node chemokines, and prevented them from secreting the immune-suppressive cytokine, IL-10. Conclusions: The number and phenotype of nDC are suppressed in advanced cancer patients. Inhibition of p38 MAPK can restore the function of patient cDC2 subset. Coculture of these p38-inhibited cDC2 with pDC further show promising results important for developing advanced DC vaccines. Our study has paved the way for a phase I clinical trial of adoptive transfer of p38-MAPK inhibited cDC2 in 24 advanced cancer patients. Legal entity responsible for the study: University of Nottingham. Funding: University of Nottingham. Disclosure: All authors have declared no conflicts of interest.
5PD
The role of EGFR inhibitor (EGFRi) in immune cell infiltration and CD81 T-cell activation in EGFR mutant lung cancer
F. Li1, G. Lizee1, P. Hwu1, X. Du2, L. Deng3, A. Talukder1, A. Katailiha1, Q. Zou3, J. Roszik1, D. Hawke1, K. Jackson1, S. Bradley1, Y. Wang3, R. Ataullakhanov4, A. Bagaev4, N. Kotlov4, V. Svekolkin4, N. Miheecheva4, F. Frenkel4, H. Sonnemann1 1 Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA, 2Oncology Department, Tianjin Beichen Hospital, Tianjin, China, 3Research and Development Department, Tianjin HengJia Biotechnology Development Co., Ltd., Tianjin, China, 4BostonGene Corporation, BostonGene Corporation, Boston, MA, USA Background: Oncogenic MAP kinase pathway activating mutations have been shown to drive immune suppression in melanoma and colorectal cancers. In this study, we explored whether oncogenic EGFR mutations play an analogous role in non-small cell lung cancer (NSCLC), and whether EGFRi can relieve tumor-associated immune suppression. Methods: Lung cancer cell lines expressing wild-type or mutated EGFR were treated with EGFRi, followed by collection of RNA and cell supernatants. In parallel, tumor biopsies from NSCLC patients receiving a personalized peptide vaccine with or without concurrent EGFRi treatment were obtained. RNAseq-based transcriptome profiles of tumor cell lines and patient tumor biopsies were compared to assess common gene signatures driven by EGFRi. Gene expression changes were confirmed at the protein level using Western blot, flow cytometry, and cytokine/chemokine Luminex. The impact of EGFRi on T-cell migration and tumor cell recognition by antigen-specific CD8þ T cells was also assessed.
v2 | Basic Science
Results: In addition to downregulating genes associated with cell proliferation, apoptosis and survival, EGFRi increased the transcription of genes associated with TNFa and TRAIL signaling, and antigen presentation. HLA class I protein upregulation was confirmed and correlated with increased recognition of tumor cells by cytotoxic T cells. Several chemokines and cytokines were up- or down-regulated following EGFRi treatment, and Luminex analysis confirmed changes to 10 of them. Migration assays demonstrated that chemotaxis of T cells towards EGFR-mutant cell supernatants increased in an EGFRi dose-dependent manner. Transcriptome profiling of tumor biopsies revealed similar gene expression changes in on-EGFRi treatment tumor samples. Furthermore, increased tumor immune cell infiltration observed in EGFRi-treated patient tumors was consistent with the upregulation of EGFRi signature chemokines at the tumor site. Conclusions: These results provide evidence that EGFRi has the capacity to facilitate modulation of the tumor microenvironment to favor immune cell infiltration and promote T-cell mediated antitumor immunity. Legal entity responsible for the study: The University of Texas MD Anderson Cancer Center. Funding: Tianjin HengJia Biotechnology Development Co., Ltd. Disclosure: F. Li: Shareholder/Stockholder/Stock options: Tianjin HengJia Biotechnology Development Co., Ltd.. G. Lizee: Advisory/Consultancy: HengJia Neoantigen Biotechnology (Tianjin) Co., Ltd.. P. Hwu: Advisory/Consultancy: Dragonfly Therapeutics; Advisory/Consultancy: GlaxoSmithKline; Advisory/Consultancy, Shareholder/Stockholder/Stock options: Immatics; Advisory / Consultancy: Sanofi; Research grant/Funding (institution): Genentech. L. Deng: Full/Parttime employment: Tianjin HengJia Biotechnology Development Co., Ltd.. Q. Zou: Full/Part-time employment: Tianjin HengJia Biotechnology Development Co., Ltd.. Y. Wang: Full/Part-time employment: Tianjin HengJia Biotechnology Development Co., Ltd.. R. Ataullakhanov: Full/Parttime employment: Bostongene. Llc. Bagaev: Full/Part-time employment: Bostongene. Llc. N. Kotlov: Full/Part-time employment: Bostongene. Llc. V. Svekolkin: Full/Part-time employment: Bostongene. Llc. N. Miheecheva: Full/Part-time employment: Bostongene. Llc. F. Frenkel: Full/Parttime employment: Bostongene. Llc. All other authors have declared no conflicts of interest.
6PD
The mutational signature of spontaneously developing tumours in MLH1-/- mice: Potential consequences for immunotherapeutic approaches
C. Maletzki1, Y. Gladbach2, M. hamed2, C. Junghanss1 Hematology, Oncology, Palliative Medicine, Universit€ atsmedizin Rostock, Rostock, Germany, 2Rostock University Medical Center, Institute for Biostatistics and Informatics in Medicine and Ageing Research (IBIMA), Universit€ atsmedizin Rostock, Rostock, Germany
1
Background: MLH1 knock out mice develop mismatch repair deficient (MMR-D) neoplasias spontaneously and reflect the diverse clinical presentation of MMR-Ddriven carcinogenesis in men. The tumor spectrum includes a high prevalence of early Non-Hodgkin T cell lymphomas (NHL), lymphoid skin lesions as well as later developing epithelial tumors of the gastrointestinal tract (GIT). Methods: Using whole-exome sequencing on MLH1-/- tumors (2x GIT, 1x splenic NHL, 1x skin lymphoma) as well as GIT-derived cell lines (n ¼ 2), we focused on identification of (I) shared and (II) mutually exclusive mutations and described the processes of ongoing mutational events in tumor-derived cultures. Results: MLH1-/- tumors show high tumor mutational burden with 3/4 primary tumor samples even being ultra-hypermutated (> 100 mut/MB). Missense mutations were more frequent than nonsense mutations, base changes were mainly due to transitions (C>T; A>G). The resulting mutational landscape was heterogeneous and in accordance with the human counterpart, MLH1-/- tumors frequently harbor mutations in PIK3CA, EGFR, KRAS, and/or ERBB3. Of note, only a few shared mutations were detectable among different tumor entities, among them were ARID1A and IDH2. Mutations in classical tumor suppressor genes SMAD4 and POLE were mutually exclusive in lymphomas, most likely contributing to a more aggressive in vivo phenotype. Comparing the mutational profile of selected tumors and their corresponding cell line revealed continuous increased numbers of somatic gene mutations. The same was true for coding microsatellite mutations in MMR-D target genes. Partial overlap was detectable, yet recognizing shared antigens. Two promising candidates are AKT3, a RACgamma serine/threonine-protein kinase and the endonuclease ERCC5 (Excision Repair 5), involved in DNA excision repair. Conclusions: The present study is the first reporting results of a comparison between different spontaneously developing tumors as models for MMR-D driven tumorigenesis. Additionally to identifying ARID1A as causative mutation hotspot, this comprehensive characterization of the mutational landscape may be a good starting point to predict antigens for vaccination approaches. Legal entity responsible for the study: The authors. Funding: german research foundation (DFG); grant number: MA5799/2-1. Disclosure: All authors have declared no conflicts of interest.
Volume 30 | Supplement 5 | October 2019
Downloaded from https://academic.oup.com/annonc/article-abstract/30/Supplement_5/mdz238.003/5576228 by guest on 24 October 2019
4O
Annals of Oncology