Browsing by Subject "RCC"

Cancer immunotherapy has advanced with the introduction of anti-PD-1 therapy. Anti-PD-1 therapy blocks the inhibitory immune checkpoint receptor PD-1, and allows T cells to regain their cytotoxic potentials in response to cancerous cells. Amongst the cytotoxic lymphocytes other than T cells, natural killer (NK) cells have not been fully explored regarding anti-PD-1 treatment. With this in mind, our aim was to characterize the immunophenotype of the immune cells in patients diagnosed with renal cell carcinoma (RCC), and separate their corresponding tumor infiltrating lymphocytes (TILs). Altogether, we analyzed the immunophenotype of the T and NK cells and RCC tumor cells from 13 RCC patients using flow cytometry. In addition, we tested the function of NK cells against RCC cells using various cell line models (NK-92 and RCC cell lines), and monitored the cytotoxicity in real time using a cell impedance assay. Based on the results, we suggest that patients diagnosed with RCC possess unique immune profiles depending on the individual, but also share characteristics regarding the tumor. In addition, NK cells have the potential to activate a cytotoxic response to RCC cells. By phenotyping the TILs, potential novel biomarkers for immuno-oncological (IO) therapy could be detected, which could aid in determining which patients would most likely benefit from immunological therapies such as anti-PD-1 treatment.

Tumors contain variable number of different immune cells that infiltrate the tumor microenvironment, such as tumor infiltrating lymphocytes (TILs). More research is needed to understand the functional and clinical importance of various TIL subgroups in cancer. Understanding the differences between individual cancer patients will help development of new treatment methods and discovering why only some patients respond to immunological treatments. Renal cell carcinoma (RCC) is the most common kidney cancer type with good overall survival prognosis when the tumor is surgically removed before it has metastasized. However, the prognosis of RCC is significantly decreased when the cancer has spread. The aim of this master’s thesis project was to characterize the tumor infiltrating lymphocyte populations in patient derived RCC samples. Characterization was done with flow cytometry and a custom antibody panel designed to detect various lymphocyte subpopulations. We also wanted to further study the TILs by expanding the lymphocytes from the tumor samples and test their function in an impendence-based assay against matched autologous tumor cells. Based on the flow cytometry results, the different RCC subtypes in the cohort showed some variation in TILs. Still, more research is needed to investigate these differences. We were able to culture the TILs from the RCC tumor samples, and most of them were CD4+ T cells expressing memory markers CD45RO and CCR7. Some expanded TILs expressed markers related to T cell activity and terminal differentiation. In conclusion, this thesis provided material and insights for future RCC TIL experiments as well as considerations for optimization needed in further studies.

Njurcancer är bland de tio vanligaste cancersjukdomarna och påträffas oftast i åldersgruppen 50+. En del former av njurcancer associeras med nedärvda syndrom, exempelvis med Von Hippel-Lindaus sjukdom. Den vanligaste typen är klarcellig njurcancer. Den molekylära bakgrunden till njurcancer ligger i störningar i gener som kodar proteiner, vilka ansvarar för blodkärlsförsörjning, metastasbildning och tillväxt av tumörcellen. Medicinsk behandling av metastaserad njurcancer ämnar hämma intracellulära signaleringsrutter, vilket kallas målinriktad vård. Vården av metastaserad njurcancer har det senaste decenniet utvecklats och ett läkemedel som introducerats inom området är kabozantinib, en multireceptortyrosinkinashämmare. Kabozantinib inhiberar tumörcellens blodkärlsförsörjning och tillväxt via blockering av flera signaleringsrutter. Syftet med studien är att undersöka vilka biverkningar rapporterades på Helsingfors universitetssjukhus onkologiska klinik av de patienter som fick kabozantinib som första linjens medicinska behandling i spridd njurcancer och ifall detta överensstämmer med tidigare forskning. Forskningen är en observationsstudie/registerstudie. Datainsamlingen skedde via patientdatasystemet och innefattar patienter vårdade 2019-2020. Data analyserades med Excel. Eftersom den slutliga patientmängden efter tillämpning av inklusionskriterier var 16, kunde ingen avancerad statistiskt hållbar analys göras. Studiens resultat var överens med tidigare forskning – de vanligaste biverkningarna var diarré och trötthet. Vidare forskning med större patientmängd kunde ge statistiskt pålitliga resultat och det vore intressant att utreda ifall fler biverkningar associeras med bättre vårdresultat. (205 ord)

Within the field of cancer immunotherapy, immune checkpoint inhibitors have been a revolution since they provoke re-activation of T-cell immune responses towards cancer. Despite their success, they only work in 13% of the patients because of a poorly immunogenic tumor, mostly due to weak T-cell infiltration. Oncolytic viruses have shown the ability to work in synergy with checkpoint inhibitors because of their tumour-specific tropism, innate immunogenicity and ability to secrete immunostimulatory agents into the tumor microenvironment. Regardless of the great potential, we lack suitable pre-clinical models to test this effect. In this study we developed renal cell carcinoma-derived organoids as in vitro platforms due to their high pre-clinical predictability compared to that of murine and in vitro 2-dimensional cell culture models. To test the ability of oncolytic viruses to stimulate the immune system, we generated three cytokine-expressing (CXCL9, CXCL10 and IL-15) oncolytic adenoviruses using a novel cloning method that we developed. We have shown that these viruses successfully produce high amount of the cytokine and attract peripheral blood mononuclear cells freshly isolated from Buffy coats. Genetically modified oncolytic adenoviruses were also shown to infect and kill human renal cell carcinoma organoids. Together, our results demonstrate the potential of organoids as test platforms for oncolytic virus -based therapy and the importance of adequate cytokine expression in T-cell recruitment. The tumor organoid platform we developed will be useful for advancing patient-specific treatment strategies and serve as a base for innovative immunotherapy models.