Charlotte O'Brien Gore
Academic and Work Experience Prior to Sept 2021 Programme Start
I obtained my Bachelor’s degree in Biology at The University of Bristol, gaining research experience in a range of areas, including trypanosome parasites in Prof Wendy Gibson’s lab and type 1 diabetes in Prof Kathleen Gillespie’s lab.
After briefly considering a career in nutritional sciences, I chose to pursue my interest in autoimmune diseases by undertaking a Master’s in immunology at King’s College London. During my MSc, I worked in Prof Leonie Taams’s lab, investigating IL-17-producing cell types in spondylarthritis.
Following my MSc, I took up a research assistant position in Dr Sophie Papa’s lab, investigating T cell exhaustion in patients with non-small cell lung cancer, through the use of CyTOF.
PhD Programme- Year 1- MRes and Project Rotations
The first year of the PhD programme involved three rotation projects:
1st rotation (Dr Joana Neves lab): Investigated the effects of bone morphogenetic protein 2 (TGF-Beta superfamily protein) and chenodeoxycholic acid (bile acid) on the differentiation of goblet cells. Methods: ilium organoid generation, cell culture, RNA extraction, cDNA synthesis, rt-qPCR, live cell imaging.
2nd rotation (Dr Luis Apolonia and Prof Mike Malim): Developed induced pluripotent stem cell-derived macrophages that specifically target CD19-expressing cancer cells through transducing them with a CD19-targeting chimeric antigen receptor. Methods: cloning, cell culture, flow cytometry, live cell imaging.
3rd rotation (Dr Alessandra Vigilante): Investigated the causes of congenital heart birth defect through RNA and single cell sequencing and bioinformatic analysis (R) of transcriptional changes in progenitor cells of patients with congenital heart disease. Methods: R programming
PhD Programme- Years 2 to 4 - Doctoral Studies
For my PhD, I chose to continue the project I undertook during my second rotation, working on the development of chimeric antigen receptor (CAR)-macrophages.
The generation of CAR-T cells has proven an effective immunotherapy for the treatment of haematological cancers but have not been as efficacious in the treatment of solid cancers. This is largely due to difficulties in CAR-T cell trafficking to- and infiltration into the tumour and in maintaining the persistence of the CAR-T cells within the highly complex tumour microenvironment.
Engineering CAR-macrophages presents a potential solution to these hindrances as macrophages are actively trafficked to the tumour site, can deploy powerful phagocytic activity, and can educate surrounding immune cells to increase the response against the tumour. Generating CAR-macrophages from primary macrophages is a long-standing challenge, owing to the macrophage’s innate and potent enzymatic biology which disrupts nucleic acid integrity, making transduction a difficult task. Generating CAR-macrophages from induced pluripotent stem cells (IPSC) can circumvent this challenge as IPSC are more easily genetically manipulated and are an exhaustless resource of cells.
The aim of my PhD project is to develop CAR-macrophages from IPSC (CAR-iMacs), that target the solid tumour antigen HER2. The differentiation of the IPSCs will be performed using a previously established protocol and the CAR design, creation and transduction into IPSC will be performed using molecular cloning techniques. This project will also include polarising the CAR-iMacs towards an M1 phenotype, to ensure an efficient inflammatory response by the CAR-iMacs towards the HER2-expressing cancer cells. This will be performed by treatment with IFNγ and other M1-inducing cytokines.
It is anticipated that the HER2-targeting CAR-iMacs will induce a potent anti-cancer effect against HER2-expressing cancer cells, ultimately contributing towards the expanding knowledge base of CAR-based solid tumour therapies.
