95. Jahresversammlung der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie

Prof. Dr.

Chairs

Prof.

NUR Wissenschaftliches Programm/ ONLY Scientific Program

Fecal Microbiota Transplantation in Blood Cancer and Results of the MAST Trial

Professor

Invited lecture

Neutrophils Link Inflammation and Microbiome in HNC

Prof. Dr. med.

Analysis of the local microbiota and the associated immunological tumor microenvironment in patients with oral and oropharyngeal squamous cell carcinoma

Analyse der lokalen Mikrobiota und des assoziierten immunologischen Tumormikromilieus bei Patienten mit oralen und oropharyngealen Plattenepithelkarzinomen

For head and neck tumors, primarily descriptive analyses of the local microbiome are available to date, which do not allow any conclusions to be drawn about the relationship between changes in the microbiota composition and carcinogenesis. In this study, the local microbiota was analyzed in patients with oral and oropharyngeal squamous cell carcinoma, and the associated immunological tumor microenvironment was also investigated. In 72 patients with oral and oropharyngeal squamous cell carcinomas, the microbial colonization of the tumor and the healthy opposite side was analyzed using 16s rRNA sequencing. The immunological tumor microenvironment was examined histologically in associated tumor tissue samples. The microbial colonization of the tumor tissue differs significantly from the contralateral healthy side. In addition to an increase in bacterial alpha diversity in the tumor tissue, individual bacterial genera are also significantly more frequently represented, primarily streptococci, fusobacteria, prevotella and porphyromonas. The colonization with streptococci, fusobacteria and prophyromonas showed a significant negative correlation with the density of neutrophil granulocytes in the tumor tissue. Oral and oropharyngeal squamous cell carcinomas have a microbial signature that differs from healthy tissue and is associated with immunological changes in the immunological microenvironment. A detailed mechanistic ex vivo analysis of the interaction of overexpressed bacterial genera with tumor cells and neutrophil granulocytes is currently underway.Für Kopf-Hals-Tumoren liegen bislang vorrangig deskriptive Analysen zum lokalen Mikrobiom vor, welche keine Rückschlüsse zum Zusammenhang zwischen Veränderung der Mikrobiota-Komposition und der Karzinogenese zulassen. In dieser Studie wurde die lokale Mikrobiota bei Patienten mit oralen und oropharyngealen Plattenepithelkarzinomen analysiert, zudem erfolgte eine Untersuchung des assoziierten immunologischen Tumormikromilieus. Bei 72 Patienten mit oralen und oropharyngealen Plattenepithelkarzinomen wurde mittels 16s rRNA-Sequenzierung die mikrobielle Besiedelung des Tumors und der gesunden Gegenseite analysiert. In assoziierten Tumor-Gewebeproben wurde das immunologische Tumormikromilieu histologisch untersucht. Die mikrobielle Besiedelung des Tumorgewebes unterscheidet sich signifikant von der gesunden Gegenseite. Neben einer Erhöhung der bakteriellen Alpha-Diversität im Tumorgewebe sind auch einzelne Bakteriengattungen signifikant häufiger vertreten, vorrangig Streptokokken, Fusobakterien, Prevotella und Porphyromonas. Für die Besiedelung mit Streptokokken, Fusobakterien und Prophyromonas zeigt sich eine signifikante negative Korrelation mit der Dichte neutrophiler Granulozyten im Tumorgewebe. Orale und oropharyngeale Plattenepithelkarzinome weisen eine mikrobielle Signatur auf, die sich vom gesunden Gewebe unterscheidet und mit immunologischen Veränderungen des immunologischen Mikromilieus assoziiert ist. Aktuell erfolgt eine detaillierte mechanistische ex vivo Analyse der Interaktion überexprimierter Bakteriengattungen mit Tumorzellen und neutrophilen Granulozyten.Die Autorinnen/Autoren geben an, dass kein Interessenkonflikt besteht

Analysis of the local microbiota and the associated immunological tumor microenvironment in patients with oral and oropharyngeal squamous cell carcinoma

Experimentelle Onkologie

Kopf-Hals-Onkologie

Free lecture

PD Dr. med.

Evaluation of primary tumor and metastasis microbiome in HNSCC

Evaluation des Mikrobioms von Primarius und Metastase bei HNSCC

Introduction Local bacterial contribution to the development of HNSCC has received increasing attention over the past few years. This study aims to investigate bacterial involvement in metastasis as a promising approach to better understand the tumor microenvironment as well as the process of metastasis. Methods Tissue samples from 24 patients with HNSCC were collected prospectively. The local ethics committee approved the study, and all patients gave informed prior consent. Qualitative microbiome analysis was performed by amplifying and sequencing the V3-V4 region of the 16S gene with the Illumina MiSeq system. Bioinformatical analysis of the sequencing data was performed with IMNGS2, Rhea and namco. Quantitative analysis of bacterial load was performed using real-time qPCR targeting the V3 region of the 16S gene. Results Quantitative PCR showed a significant difference between the bacterial load of the lymph node metastasis and that of the negative controls for contamination. In the qualitative analysis of the bacterial signatures with 16S sequencing, mainly bacteria of the strains Proteobacteria and Firmicutes were found in the tumor samples. In the individual comparison of primary tumor and metastasis of the same patient, it can be determined that bacterial strains found in the metastasis are also predominantly found in the primary tumor. Conclusion Bacteria are present in lymph node metastasis of HNSCC. However, primary tumor and metastasis differ in their beta diversity. A possible explanation for this would be a shift of the microbiome due to differences in the microenvironment. To answer the question to what extent bacteria affect the tumor microenvironment and metastasis, further investigations are needed. Die Autorinnen/Autoren geben an, dass kein Interessenkonflikt besteht.

Evaluation of primary tumor and metastasis microbiome in HNSCC

Interactions of myeloid cells with tumor microenvironment as a therapeutic target in head and neck cancer

Immune evasion of cancer is induced by interaction between immune cells with tumor microenvironment (TME). CCL2 is a major chemoattractant for myeloid cells in TME and is associated with immunosuppression at the tumor site and poor prognosis. Aiming to prevent interaction of myeloid cells with immunosuppressive TME, we developed a peptide fragment of CCL2, MMIb, which blocks glycosaminoglycan-CCL2 interaction and thus disturbs the guiding chemokine gradient for myeloid cells. In translational approach, the effect of MMIb was evaluated in vitro and in vivo in murine model of oropharyngeal carcinoma, and verified in human in vitro system using head and neck cancer (HNC) explants. Molecular mechanism was investigated using LC-MS/MS and confirmed by multiparameter flow cytometry. Treatment with MMIb prevented infiltration of tumors with myeloid cells and suppressed tumor growth in vivo. Analysis of tumor proteome after MMIb treatment revealed upregulation of the pathways responsible for antigen uptake, processing and presentation via MHCI and MHCII, immunostimulation (PDL1-, TGFb-, IL12+) and lymphocyte activation (PD1-, Fas+, GrzmB+). Moreover, MMIb prevented tumor-induced immunosuppression in tumor-draining lymph nodes. In human system, MMIb suppressed migration of labeled monocytes into the HNC explants and their immunosuppressive polarization. Altogether, we can demonstrate the efficiency of MMIb to therapeutically prevent immune cell/TME interactions and therefore to maintain their anti-tumor potential. Importantly, the effect is not limited to one molecule or one cell type, but covers the broad range of immunoregulatory mediators, making it less sensitive to immune evasion. We believe that this compound can be considered as immunosupportive medication in cancer.The authors declare that there is no conflict of interest

Interactions of myeloid cells with tumor microenvironment as a therapeutic target in head and neck cancer

PD Dr.

Myeloid derived supressor cells contribute to distinct immune profiles in HPV+ and HPV- head and neck cancer patients

Introduction: Patients with HPV- localized head and neck cancer show distinct therapeutic responses compared to HPV-associated cancers, implicating differences in immune status and immune response. Therefore, we analyzed immune profiles of myeloid-derived suppressor cells (MDSC) in HPV+ versus HPV- disease and their influence on CD8+ T cells. Material and methods: Circulating Immune cells and Tumor infiltrating cells were investigated in two patient cohorts (n=89/ n= 61) via flow cytometric evaluation, ELIspot and immunohistochemistry. Results: Circulating monocytic MDSC were increased in patients with HPV− disease, associated to lower fractions of terminally differentiated CD8+ effector cells. HPV− tumors displayed lower infiltration rates of CD8+ and CD45RO+ immune cells compared with HPV+ tumors. Importantly, frequencies of tumor tissue-infiltrating PMN-MDSC were increased, while percentages of Granzyme B+ and Ki-67+ CD8 T cells were reduced in patients with HPV− disease. Within the HPV+ group, patients exhibiting low levels of intratumoral CD66b+/LOX1+ PMN-MDSC showed significantly improved survival rates. Conclusion: We observed differences in frequencies and relative ratios of MDSC and effector T cells in HPV− HNC compared with more immunogenic HPV-associated disease. Our data provide new insight into the immunological profiles of these two tumor entities and may be utilized for more tailored immunotherapeutic approaches in the future.Die Autorinnen/Autoren geben an, dass kein Interessenkonflikt besteht.

Myeloid derived supressor cells contribute to distinct immune profiles in HPV+ and HPV- head and neck cancer patients

Phase I clinical study of intralesional TLR-7-Agonist (LHC165) with PD-1 inhibitor Spartalizumab in Patients with advanced PD1-resistant solid tumors

Phase I klinische Studie eines intratumoral injizierten TLR7-Agonisten (LHC165) mit dem PD1-inhibitor Spartalizumab in Patienten mit fortgeschrittenen PD1-resistenten soliden Tumoren

Background. Especially for patients with cisplatin- and anti-PD-1 resistant advanced head and neck squamous cell cancer (HNSCC), therapeutic options are very rare. For this highly palliative cohort additional therapeutic approaches are urgently needed. In pre-clinical studies, the toll like receptor 7 agonist LHC165 has demonstrated a strong anti-tumor effect in advanced solid cancers. Methods. Our patient presented with cisplatin- and anti-PD1-resistant advanced HNSCC at the floor of the mouth. He was included in the phase I/Ib dose expansion study of LHC165 in combination with the PD-1 inhibitor spartalizumab (NCT03301896, sponsored by Novartis). LHC165 was administered biweekly by intratumoral injection (n=4) over six months. Spartalizumab was administered intravenously every four weeks. Results. The patient has shown durable complete clinical response (&gt;3 years). After termination of the study, treatment was continued with pembrolizumab 200mg i.v. every three weeks. Oral nutrition is well feasible without the need for a percutaneus gastral feeding tube. In the dose-escalation phase of the study (n=39), overall response was partial response (PR, 3/39, 8 %) and stable disease (SD, 4/39, 10 %). Immune monitoring tended to show increased intratumoral levels of CD8 and CD68 cells in patients with PR or SD. Conclusion. Intralesional LHC165 in combination with spartalizumab demonstrated an exceptional clinical response in a patient with advanced HNSCC. Thus, the combination of TLR7-agonist in combination with i.v. PD1-inhibitor can be a treatment option for a selected group of palliative cancer patients.Hintergrund. Insbesondere für Patienten mit Cisplatin- und anti-PD-1-resistentem fortgeschrittenem Kopf-Hals-Plattenepithelkarzinom (HNSCC) gibt es nur noch wenige Therapieoptionen. Für diese hoch palliative Kohorte sind zusätzliche Therapieansätze dringend erforderlich. In präklinischen Studien hat der Toll-like-Rezeptor-7-Agonist LHC165 eine starke Antitumorwirkung bei soliden Krebsarten gezeigt. Methoden. Unser Patient stellte sich mit Cisplatin- und anti-PD1-resistentem, fortgeschrittenem HNSCC am Mundboden vor. Er wurde in die Phase-I/Ib-Dosiserweiterungsstudie von LHC165 in Kombination mit dem PD-1-Inhibitor Spartalizumab (NCT03301896, gesponsert von Novartis) eingeschlossen. LHC165 wurde über einen Zeitraum von sechs Monaten alle zwei Wochen durch intratumorale Injektion (n=4) verabreicht. Spartalizumab wurde alle vier Wochen intravenös verabreicht. Ergebnisse. Der Patient zeigte ein dauerhaftes vollständiges klinisches Ansprechen (&gt;3 Jahre). Nach Beendigung der Studie wurde die Behandlung mit Pembrolizumab 200 mg i.v. fortgesetzt (Q3W). Die orale Ernährung ist gut möglich, ohne dass eine perkutane Magensonde erforderlich ist. In der Dosissteigerungsphase der Studie (n=39) zeigten die Patienten ein partielles Ansprechen (PR, 3/39, 8 %) und eine stable disease (SD, 4/39, 10 %). Die Immunüberwachung zeigte bei Patienten mit PR oder SD tendenziell erhöhte intratumorale Spiegel von CD8- und CD68-Zellen. Abschluss. Die intratumorale Injektion von LHC165 in Kombination mit Spartalizumab zeigte ein außergewöhnlich gutes klinisches Ansprechen bei einem Patienten mit fortgeschrittenem HNSCC. Somit kann die Kombination eines TLR7-Agonisten in Kombination mit i.v. PD1-Inhibition eine Behandlungsoption für eine ausgewählte Gruppe in der Palliativsituation darstellen.Die Autorinnen/Autoren geben an, dass kein Interessenkonflikt besteht.

Phase I clinical study of intralesional TLR-7-Agonist (LHC165) with PD-1 inhibitor Spartalizumab in Patients with advanced PD1-resistant solid tumors

International Session 25
Microbiome and Inflammation

Termin

Chairs

Thema

Sprache

Programm

International Session 25Microbiome and Inflammation

Termin

Chairs

Thema

Sprache

Programm

International Session 25
Microbiome and Inflammation