Charakterisierung und funktionelle Analyse von tumorantigen-spezifischen Immuninfiltraten bei Patienten mit Kopf-Hals-Karzinomen
Zusammenfassung der Projektergebnisse
I was able to find with the first scRNA Seq results differences between the TILs in the primary and the draining lymph node metastasis. In our first preliminary data, we saw the differences especially in the cytotoxic function such as granzyme. Also, the scTCR seq revealed that TRMs were in particular the cell type with a high expansion rate, which were also found in the tumour and the tissue with the same TCR. The whole dataset, which was analysed by whole RNA Seq, was used for MiXCR TCR analysis which also found a high abundancy of shared clonotypes between the tumour sites. As this cohort consists only of HPV positive cases, we assume those share clonotypes a most likely against HPV antigens or less likely against some neoantigen/shared antigen which was already in the primary and travelled along to the lymph node with the metastasis. To address this question is to stimulate the blood of those patients for HPV antigens and run an scRNA TCR Seq or whole TCR Seq to assess whether those TCRs can be found in the tumour sites as well. I have learned this TCR analysis which was performed with a bioinformatics R package called immunarch and was involved in several different projects for this type of analysis. In a multi-site cancer (neuroendocrine tumour) we were able to detect sharing of the clonotypes between different lymph node metastasis and also found overlap with neoantigen stimulated CD4+ and CD8+ T cell cultures. These data have an importance for a fundamental understanding of T cell behavior as well as impacting our thinking about personalized cancer vaccine development. Beside the neoantigens, cancer testis antigens, shared antigens and viral antigens (such as HPV) are important as possible vaccination targets. This interesting topic was summarized by me and the group in a review. Additionally, we were able to assess the TRMs in primary, lymph node metastasis and recurrence tissue with my developed HNSCC TMA and MxIHC, were we saw survival benefit of the TRM in primary tissue but not in the lymph node or recurrence tissue. Interestingly, the HPV status has a strong effect on the TRM counts in a tumour as we know. However, this difference between the HPV positive and negative tumours couldn’t be seen in the lymph node metastasis anymore. A coinhibitory receptor, TIM3, the expression of which our group had reported to ‘tag’ particularly protective CD8+ T cells in lung cancer, was expressed in HNSCC also, with higher levels on TRM and non TRM in the lymph node metastasis tissue and the recurrence tissue. Our data point to a potential role of targeting TIM3 in patients with nodal disease or when they relapse and warrant further prospective evaluation. Lastly the analysis of patient’s serum for HPV16 E2 and E7 antibody reactivities revealed a survival benefit for E2 IgG but not for IgA. Also, we found the connection between viral antigen presence in the tumour and the reactivity in the blood. Which was then also correlated to TIL status, which closes the loop of adaptive and humoral immune response.
Projektbezogene Publikationen (Auswahl)
- HNSCC: Tumour Antigens and Their Targeting by Immunotherapy. Cells. 2020;9(9)
von Witzleben A, Wang C, Laban S, Savelyeva N, Ottensmeier CH
(Siehe online unter https://doi.org/10.3390/cells9092103) - Immune checkpoint expression in HNSCC patients before and after definitive chemoradiotherapy. Head Neck. 2020
Doescher J, Minkenberg P, Laban S, Kostezka U, von Witzleben A, Hoffmann TK, et al.
(Siehe online unter https://doi.org/10.1002/hed.26534) - Peripheral Cytokine Levels Differ by HPV Status and Change Treatment-Dependently in Patients with Head and Neck Squamous Cell Carcinoma. Int J Mol Sci. 2020;21(17)
Mytilineos D, Ezic J, von Witzleben A, Mytilineos J, Lotfi R, Furst D, et al.
(Siehe online unter https://doi.org/10.3390/ijms21175990) - Prospective longitudinal study of immune checkpoint molecule (ICM) expression in immune cell subsets during curative conventional therapy of head and neck squamous cell carcinoma (HNSCC). Int J Cancer. 2020
von Witzleben A, Fehn A, Grages A, Ezic J, Jeske SS, Puntigam LK, et al.
(Siehe online unter https://doi.org/10.1002/ijc.33446) - Correlation of HPV16 Gene Status and Gene Expression with Antibody Seropositivity and TIL Status in OPSCC. Frontiers in Oncology. 2021;10(3049)
von Witzleben A, Currall E, Wood O, Chudley L, Akinyegun O, Thomas J, et al.
(Siehe online unter https://doi.org/10.3389/fonc.2020.591063)
