Innovative tools to detect Glioblastoma (GBM) exosomes

Informations projet

Gl.EXO

N° de convention de subvention: 891551

DOI

10.3030/891551

Projet clôturé

Date de signature de la CE 3 Avril 2020

Date de début 1 Juillet 2020

Date de fin 30 Juin 2022

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 175 572,48

Contribution de l’UE

€ 175 572,48

Coordonné par

PERCUROS BV
Netherlands

Periodic Reporting for period 1 - Gl.EXO (Innovative tools to detect Glioblastoma (GBM) exosomes)

Période du rapport: 2020-07-01 au 2022-06-30

Résumé du contexte et des objectifs généraux du projet

Glioblastoma is the most frequent and aggressive primary brain tumour in adults (Louis et al., 2016; Sant et al., 2012). Despite advances in surgical and medical neuro-oncology, prognosis for GBM patients remains dismal, with a median survival of 14–15 months (Urbanska et al., 2014). A small population of cancer stem cells (glioblastoma stem cells, GSCs) that retains stem cell properties, including self-renewal and multipotency, are deemed responsible for the frequent relapse of GBM and its resistance to conventional therapeutics (Bao, S. et al. 2006; Bovenberg, M.S. 2013). Despite the important advancements in the treatment of GBM the clinical outcome of patients is still disappointing. Moreover, the traditional diagnostic methods are limited by the tumour tissue accessibility. Therefore, the research of circulating biomarkers is a fundamental challenge that can allow detecting tumours early, when they are most treatable, consequently improving disease outcome and health costs. Indeed, A40s-based assay could be crucial for an easily and early-stage diagnosis of GBM along with a quick and always available tool for monitoring GBM treatments and possible tumour relapse. This will give a better treatment of patients with great impact both on patients’ overall survivor and quality of life and on health systems economics.
Gl.EXO made important advances in pursuing 3 objectives:
The first objective was to characterize A40s aptamer binding on blood exosomal samples. Indeed, exosome-coated plates have been generated and used with aptmer-based reporter system to convert target binding into a detectable signal. Different protocols have been tested with the aim of selectively discriminating positive to negative exosomes from serum and the possibility to increase assay sensitivity through an additional different aptamer.
The second objective was to evaluate the clinical potential of A40s by analysing its ability to bind tumoral exosomes from human serum. Indeed, A40s has been tested on a bigger cohort of patients with different methods. Our goal was to demonstrate that A40s is able to be translatable into the clinical practice for detecting GBM exosomes in the blood of GBM patients.
The third objective was the theragnostic application of A40s and so its use as a therapeutic tool. Here, our aim was to exploit the aptamer exosomal binding properties together with its ability to inhibit exosomes functional effects. A40s has been able to affect exosomal pro-tumorigenic effects. Additionally, A40s has been proven to be able to deliver near-infrared agents usable for both imaging and photodynamic therapy, demonstrating A40s possible use as theragnostic.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

The project applied a new discovered aptamer, A40s, able to bind and discriminates them from differentiated cells by binding EphA2 receptor. Aptamer-based ELISA-like assay has been set up. Results demonstrate the possibility to use A40s during ELONA assay for detecting GSCs released exosomes. A40s aptamer has been validated for its pre-clinical sensitivity, specificity, accuracy, and precision in the early detection of GBM. Secondly, the therapeutic potential of A40s, along with its diagnostic functionality, has been also evaluated. A40s proved to be able to directly affect stem cell markers and to be used as a diagnostic tool. This allows having a diagnostic and a therapeutic effect in the same molecule.
Exploitation and Dissemination overview
These results have been shown during meetings and events (like seminars and monthly meetings), besides the publication of scientific peer-reviewed articles in high-impact journals related to aptamers, exosomes, and diagnostic, prognostic and therapeutic approaches for cancer submitted by Dr Affinito.
The following scientific articles have been published or submitted by Dr Affinito during the MSCA individual fellowship on open access journals and public repositories (Zenodo), with acknowledgment to the EU Horizon 2020 funding.
- Four research articles have been published from July 2020 by Dr Affinito (impact factors range: 5.68-27.20 with 22 total citations)
- One additional article has been accepted so far (impact factor: 6.11- topic: RNA aptamer selection).
- One systematic review has been published (impact factor: 6.64 - topic: RNA and DNA aptamers in Glioblastoma diagnosis and therapy).
Dr Affinito participates in training and conferences during the lifetime of Gl.EXO. She received scientific training related to mass spectrometry and fluorescence that increased Dr Affinito scientific knowledge along with the weekly Percuros meeting and monthly science meeting in which she acts as a moderator. These scientific trainings had been associated with non-scientific trainings concerning grant writing, patent application, marketing, gender issues and equality plan. Dr Affinito also participated to a workshop on Leadership and effective management. Additionally, during Gl.EXO project, Dr Affinito transferred her gained knowledge on aptamers as diagnostic and theragnostic to medical intern and scientific staff, and actively participated to several events. She gave a talk at the Horizon Europe Weeks meeting in Wieliczka-Krakow on June 28 to June 30, 2021, a seminar on diagnostic application through aptamers at LUMC Leiden on 14.7.2021 and a lecture related to Responsible Research and Innovation at the 1st training event of MSCA-ITN-PIANO in Davos on 04.04.22.
Additionally, the Gl.EXO project and Dr Affinito experience in Horizon 2020 have been promoted through PERCUROS BV social profiles (both Facebook and LinkedIn) and in Dr Affinito personal profiles (Facebook and Instagram), with 2606 people reached with these posts.

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

A timely, accurate and non-invasive diagnostic assay for cancer exosomes detection has been developed to have a well-timed therapeutic approach due to a rapid diagnosis as well as a fast identification of the different GBM subtypes to tailor the treatment approach. This assay uses a modified RNA-aptamer able to selectively bind circulating GBM exosomes and, thus, to provide a timely, real-time and non-invasive diagnosis of GBM.
Gl.EXO project will have a great socioeconomic impact considering that GBM diagnosis and treatment is among the most expensive tumours.

GBM diagnosis is currently based on surgery, imaging and lab analysis. Direct costs of this disease have significant differences between different institutions and countries. Craniotomy cost an average of $10,042; whereas imaging and laboratory costs range from $1,135 to $13,228. Instead, treatments for the recurrent disease have an average cost of $27,442 ± 18,992.
It’s clear the importance to have a new, not invasive and timely diagnostic tool (different from the current diagnostic instruments for GBM) both in terms of quality of life for patients and from a socio-economic point of view (Nicholas J. Goel 2021, https://doi.org/10.1080/13696998.2021.1964775).
This system will have a very high potential impact on GBM, by fitting in the stuck diagnostic and therapeutic scenario of this disease.

A Affinito