Title: Is B cell Immune phenotype predictive of outcome and toxicity with combination checkpoint blockade in Non-small cell lung cancer patients?
Funder: Cancer Research UK

Abstract: Background: Immune research in NSCLC has focused predominantly around T-cell immune-biology, however, in reality the immune response is a complex interplay between the primary tumour and microenvironment, T and B cells. Unlocking the phenotypic diversity of the B cell milieu in NSCLC will enable the identification of immune cell patterns that correlate with outcome (negative or positive) which may harbour new avenues for immunotherapy. In the field of renal cell carcinoma, distinct immune cell (T cell and macrophage) compositions were identified in 73 patients with disease and correlated with progression free survival; they developed a robust human atlas of the immune cell signature in these patients that revealed potential biomarkers for disease as well as targets for immunotherapy development. It is this paradigm upon which we aim to develop our B cell study in NSCLC patients on immunotherapy, an area which has not yet been ventured. Aims: We wish to ascertain if B cell profile influences outcome in patients with advanced disease being treated with immune-oncotherapy (IO) alone, combination IO/IO or IO with chemotherapy. Methods: Our primary experimental method will serve to determine the properties and phenotype of B cells circulating in the blood of NSCLC patients on checkpoint blockade therapy. In order to do this we will use mass cytometric analysis, the cells will be pre-labelled using antibodies that are conjugated to heavy metal ion tags which will enable readouts as a function of time of flight through the apparatus, hence the name CyTOF; Cytometry by Time of Flight. Using a pre-conceived B cell CyTOF antibody panel, we will profile the B cell immune signature in these lung cancer patients. Given the paucity of deep B cell immune-profiling in humans with lung cancer, we will base our panel on an existing panel (detailed below) used currently by our institute for work in myeloma and myeloid leukaemia. The panel has been modified to account for current data in B cell biology in the context of solid tumours and particular NSCLC. We will compare our mass cytometry panel to flow cytometric analysis in order to ensure reliability of data. Results of research: If we can identify immune cell patterns that strongly correlate with outcome and response to treatment, the B cell signature will serve to act as a biomarker and enable patient stratification into particular treatment arms to maximise therapeutic benefit in a time efficient manner.

Lay abstract: Background: Immune research in NSCLC has focused predominantly around T-cell immune-biology, however, in reality the immune response is a complex interplay between the primary tumour and microenvironment, T and B cells. Unlocking the phenotypic diversity of the B cell milieu in NSCLC will enable the identification of immune cell patterns that correlate with outcome (negative or positive) which may harbour new avenues for immunotherapy. In the field of renal cell carcinoma, distinct immune cell (T cell and macrophage) compositions were identified in 73 patients with disease and correlated with progression free survival; they developed a robust human atlas of the immune cell signature in these patients that revealed potential biomarkers for disease as well as targets for immunotherapy development. It is this paradigm upon which we aim to develop our B cell study in NSCLC patients on immunotherapy, an area which has not yet been ventured. Aims: We wish to ascertain if B cell profile influences outcome in patients with advanced disease being treated with immune-oncotherapy (IO) alone, combination IO/IO or IO with chemotherapy. Methods: Our primary experimental method will serve to determine the properties and phenotype of B cells circulating in the blood of NSCLC patients on checkpoint blockade therapy. In order to do this we will use mass cytometric analysis, the cells will be pre-labelled using antibodies that are conjugated to heavy metal ion tags which will enable readouts as a function of time of flight through the apparatus, hence the name CyTOF; Cytometry by Time of Flight. Using a pre-conceived B cell CyTOF antibody panel, we will profile the B cell immune signature in these lung cancer patients. Given the paucity of deep B cell immune-profiling in humans with lung cancer, we will base our panel on an existing panel (detailed below) used currently by our institute for work in myeloma and myeloid leukaemia. The panel has been modified to account for current data in B cell biology in the context of solid tumours and particular NSCLC. We will compare our mass cytometry panel to flow cytometric analysis in order to ensure reliability of data. Results of research: If we can identify immune cell patterns that strongly correlate with outcome and response to treatment, the B cell signature will serve to act as a biomarker and enable patient stratification into particular treatment arms to maximise therapeutic benefit in a time efficient manner.

Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS