16P Development of a 3D lung cancer model and evaluation of the efficiency of a human antibody directed against a novel target

Abstract

In-vitro cell models have been employed for the study of lung cancer progression and metastasis. Most often, these models are 2D cell cultures and are not representative of cancer complexity and cell interactions in vivo. Immune therapy, alone or in combination with chemo- or radio treatment is an attractive alternative for lung cancer management. We have designed a 3D model of lung cancer composed of bronchial epithelial, fibroblasts, epithelial, lung adenocarcinoma and immune cells grown in different layers in a matrigel. Genes upregulated in the 3D culture were evaluated and an antibody against a specific over-expressed protein produced. The biological efficacy of the produced antibody on the expression of gene and various cell markers was evaluated. Expression of cell markers was determined using flow cytometry. Antibody was produced using immune cells cultured ex-vivo and activated against an immunogenic epitope of the protein-target. Antibody efficiency was calculated using gene and flow cytometry analysis. By genetic analysis, BMPR2 was one of the genes found over-expressed and was chosen as the antibody target. Lung cancer model was found to have increased expression of markers like Epcam (over expressed in lung cancer) and Notch (promotes tumor initiation). Analysis of the immune cell population only, showed an increased in the expression of CD25 (activated T and B cells), CD80/86 (APC), CD206 (macrophages), and also CTLA4 (down regulation of immune system). Incubation of the lung model with anti-BMPR2 antibody, decreased cell growth, increased immune marker expression (CD206) and decreased expression of genes involved in cancer progression (CD44, FOS, NRAS, ARAF). We have developed a 3D cell culture model for lung cancer. We have identified a potential target for immunotherapy and developed and antibody against the specific target. It was found that addition of anti-BMPR2 antibody in our lung cancer model increased the expression of genes involved in immune activation and decreased genes involved in tumor progression. Our antibody could potentially be a novel therapeutic antibody for lung cancer treatment.