Title: Early detection of lung cancer using a molecular tracer
Funder: Cancer Research UK

Abstract: Background: This project focuses on a protein, ROR1, that is highly expressed on immune-driven aggressive tumours (lung, pancreatic, ovarian and TNBC). ROR1 expression levels correlate with poor prognosis in non-small cell lung cancer (NSCLC). It is accepted that ROR1+ tumours are particularly aggressive and so an effective strategy in lung cancer treatment would be early detection of ROR1+ malignancies. ROR1 is expressed by a small number of individual cells in healthy lungs and we believe changes in ROR1 expression will occur in early tumourigenesis. The Nathwani group investigate T cell based immunotherapies targeting ROR1 and have a library of antibodies against ROR1 (mouse and human) that have been produced as a library of single chain variable fragments (scFvs). Aims: 1. Define ROR1 expression changes during early tumourigenesis 2. Engineer existing ROR1 antibody fragments to detect changes in ROR1 expression in lung nodules with high sensitivity and specificity. Methods: To validate ROR1 as an early detection molecule we will use state-of-the-art imaging techniques in UCL Centre of Advanced Biomolecular Imaging (CABI)to allow identification and characterisation of early lung lesions in murine models using optical tomography (OPT) and high-resolution episcopic microscopy (HREM) alongside standard CT imaging. From our library we will define scFvs with the most favourable binding kinetics for in vivo detection of ROR1. After inserting scFv fragments into a larger protein backbone, we will further optimize the panel of tracer molecules using a strategy of introducing specific mutations within the backbone sequence. This process will facilitate use of these molecules as highly sensitive and specific NSCLC early detection tracers. We will subsequently conduct proof-of-principal in vivo studies with fluorescently labelled tracer molecules in mice with early NSCLC. How the results of this research will be used: A primer award will enable us to generate preliminary data for a full project award application. A future project would involve looking at tumours tissues from NSCLC patients diagnosed with early NSCLC (stage I and II) and assessing non-labelled tracer binding to tissue sections by detection of the histidine tag on the ROR1-scFv. We would further optimize our selected ROR1 binder for use as a tracer through affinity maturation and conduct proof of principal PET studies in murine NSCLC with a radiolabelled tracer. These investigations will precede initial first-in-man PET imaging studies. Our overarching objective is to improve on existing imaging techniques and screen high risk patients for molecular changes in pulmonary nodules.

Lay abstract: Background: This project focuses on a protein, ROR1, that is highly expressed on immune-driven aggressive tumours (lung, pancreatic, ovarian and TNBC). ROR1 expression levels correlate with poor prognosis in non-small cell lung cancer (NSCLC). It is accepted that ROR1+ tumours are particularly aggressive and so an effective strategy in lung cancer treatment would be early detection of ROR1+ malignancies. ROR1 is expressed by a small number of individual cells in healthy lungs and we believe changes in ROR1 expression will occur in early tumourigenesis. The Nathwani group investigate T cell based immunotherapies targeting ROR1 and have a library of antibodies against ROR1 (mouse and human) that have been produced as a library of single chain variable fragments (scFvs). Aims: 1. Define ROR1 expression changes during early tumourigenesis 2. Engineer existing ROR1 antibody fragments to detect changes in ROR1 expression in lung nodules with high sensitivity and specificity. Methods: To validate ROR1 as an early detection molecule we will use state-of-the-art imaging techniques in UCL Centre of Advanced Biomolecular Imaging (CABI)to allow identification and characterisation of early lung lesions in murine models using optical tomography (OPT) and high-resolution episcopic microscopy (HREM) alongside standard CT imaging. From our library we will define scFvs with the most favourable binding kinetics for in vivo detection of ROR1. After inserting scFv fragments into a larger protein backbone, we will further optimize the panel of tracer molecules using a strategy of introducing specific mutations within the backbone sequence. This process will facilitate use of these molecules as highly sensitive and specific NSCLC early detection tracers. We will subsequently conduct proof-of-principal in vivo studies with fluorescently labelled tracer molecules in mice with early NSCLC. How the results of this research will be used: A primer award will enable us to generate preliminary data for a full project award application. A future project would involve looking at tumours tissues from NSCLC patients diagnosed with early NSCLC (stage I and II) and assessing non-labelled tracer binding to tissue sections by detection of the histidine tag on the ROR1-scFv. We would further optimize our selected ROR1 binder for use as a tracer through affinity maturation and conduct proof of principal PET studies in murine NSCLC with a radiolabelled tracer. These investigations will precede initial first-in-man PET imaging studies. Our overarching objective is to improve on existing imaging techniques and screen high risk patients for molecular changes in pulmonary nodules.

Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: CANCER AND NEOPLASMS