Super-resolution microscopy for immune checkpoint inhibitors diagnostics

Project Information

NANODIAGNOSTIC

Grant agreement ID: 101100873

DOI

10.3030/101100873

Project closed

EC signature date 10 November 2022

Start date 1 December 2022

End date 31 May 2024

Funded under

• European Research Council (ERC)

Total cost

No data

EU contribution

€ 150 000,00

Coordinated by

TECHNISCHE UNIVERSITEIT EINDHOVEN
Netherlands

Periodic Reporting for period 1 - NANODIAGNOSTIC (Super-resolution microscopy for immune checkpoint inhibitors diagnostics)

Reporting period: 2022-12-01 to 2024-05-31

Summary of the context and overall objectives of the project

This project main objectives were to develop and set-up a complete workflow for live-cell single-molecule PAINT imaging of PD-L1 in circulating tumour cells (CTC) derived from patients as a novel diagnostic tool. This includes from the imaging probe design and labelling, isolate and capture of CTCs from liquid biopsies and analysis of the single-molecule tracking data. This project aimed at bringing an advanced microscopy technique from the laboratory setting to a useful diagnostic tool for clinics. This is very important and timing. Many new microscopy methods are entering the arena but they re generally too complex, not validated and not standardised to be used in a clinical setting. Among them super-resolution and single molecule microscopy proved to be powerful methods in the research settings but they are currently not applied in clinical setting. Super-resolution can provide imaging with high sensitivity, spatial resolution and dynamic analysis (study molecular mobility), all useful information in the clinical setting. This projects aims to provide a proof of concept of this advanced technique in the diagnostic setting.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

We tackle the challenge from all angles to meet all the needs for a super-resolution clinical measurements. In particular we work on validating: i) probes for imaging, ii) imaging workflow, iii) data analysis and iv) data mining with AI.
We designed an imaging probe by labelling a Fab fragment of a PD-L1 antibody, and tested it in relevant cell lines to determine imaging conditions. We worked on lung cancer liquid biopsies aiming to capture and analysis circulating tumour cells (CTCs). We developed a CTC isolation procedure from blood and pleural samples, as well as an antibody capturing slide to attach CTC at the surface for TIRF imaging. We have also successfully imaged CTC derived from patient utilizing labelled Fab for PD-L1 and EGFR (simultaneous dual colour) and extracted all the single-molecule tracking information. This has shown the first validation in a relevant environment of single-molecule PAINT as a diagnostic technique in a relevant environment.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The output of this project has stablished the workflow of single-molecule PAINT in live CTCs. This eases the path for complex microscopy techniques to get out of the laboratory setting into the clinics where they can make an impact. Next clear step is the demonstration of this technology potential to predict therapy outcome in patients. We have already started the IPR protection by filling in a patent. After these to next steps are complete we would design an appropriate commercialization strategy and look for financial support to achieve the market and expand.