WP2 is clearly devoted to in vivo real-time diagnosis and to the development of novel instruments. Two instruments have been developed. One is more advanced and is devoted for guiding surgery in a non-invasive manner by providing in vivo MS analysis in real-time using an infrared-based OPO-laser. The project is named Spidemass. The second one is in emergence and is devoted to real-time diagnosis of volatile organic compounds (VOCs) based on low-temperature plasma (LTP) coupled to mass spectrometry, named Snoop-I.
SPIDERMASS : Intra-operative diagnostics for oncology
Support: ANR, INCA, SiricOncoLille , SATT Nord
Partners : CHRU Lille, Comprehensive Care Center Oscar Lambret, OCR, EAL Lancet
Data collection for diagnosis and prognosis of pathologies represent critical information for the physicians and surgeons to make a decision for the subsequent step of treatment and uptake of the patients. Nowadays, tumour grading is still largely performed through microscopic examination of biopsies using histology and immunohistochemistry techniques. These examinations remain difficult especially for data reading (leading to a more or less important variability inter-exam) or are targeted to certain selected markers. These exams are complicated if the surgeons need the information during surgery. Using conventional tools, it is still remains difficult for surgeons to determine pathological tissues and excision margins. There is a need to develop a totally novel instrument usable in the operating room and allowing the collection of information in real time to obtain a fast and precise diagnosis and prognosis for a personalized treatment. SpiderMass is such novel guided surgery instrument. SpiderMass is an infrared (IR) AP MALDI MS system with a remote ion source for in-vivo real-time conditions. By laser excitation of endogenous water molecules it was demonstrated that in-vivo real-time analysis of molecular signatures from tissue surfaces, such as human skin for example, could be achieved, allowing the detection of metabolites and lipids with masses up to 2000 amu (1 amu = 1 Da = 1 u). In the SpiderMass system, an optic fiber guides the IR laser beam to a handpiece designed for manually scanning the tissue surface. The gas phase ions produced by the laser irradiation are transferred through a thin, flexible tube by aspiration to the atmospheric pressure inlet of an MS instrument, yielding a ‘spectral fingerprint’ of the tissue’s metabolome in real-time
(A) Schematic representation of the SpiderMass instrument. (B) Typical time evolution of the total ion current (TIC) recorded by the MS instrument for an irradiation period of 30s. (C) Typical MS spectrum acquired in negative mode extracted by averaging the MS spectra over the full irradiation period. (D) Optical microscope images from bovine liver tissue sections obtained after irradiation allowing determination of the maximum ablation depth. (E) Photographs of the SpiderMass prototype system showing the optical fiber guiding the laser to the handpiece for surface scanning, as well as the transfer line leading to an MS analyser (Orbitrap, Thermo Fisher Scientific).
SNOOP-I : non-invasive and real-time analysis tool for the detection of Volatile Organic Compounds (VOCs)
Support: I-Site, ULille, SATT Nord
Partners : Comprehensive Care Center Oscar Lambret, OCR
SNOOP-I aims to develop a new non-invasive and real-time analysis tool for the detection of Volatile Organic Compounds (VOCs) to enable early diagnosis of cancer and follow-up of patients during treatment. SNOOP-I is a project to develop a new non-invasive and real-time analysis tool for Volatile Organic Compounds (VOCs) to enable early diagnosis of cancer and follow-up of patients during treatment. Indeed, patient survival is closely correlated with the ability to diagnose the disease early at a stage where it remains localized. Being able to perform an early diagnosis, in a totally non-invasive way in real-time and on a large scale is therefore an important societal issue. Various studies demonstrate the ability of VOCs released by the body in the breath, urine, faeces or skin to provide highly specific and sensitive molecular signatures for cancer detection. Here we want to develop an electronic dog based on robustness, extreme sensitivity (up to ppt) and the ability to detect complex molecular signatures of mass spectrometry to obtain a real-time and large-scale analysis system ladder. The project will be organized around the development and optimization of the new instrument, then the creation of banks and the identification of the VOCs associated with cancers. Finally, the system will be tested for the analysis of VOCs originating from samples of patients, firstly canines then human.
European Associated Laboratory (EAL) LANCET
In 2017, Inserm and Imperial College of London have labialized a European Associated Laboratory (EAL), named LANCET. LANCET EAL is based on two groups, PRISM and the Department of Surgery and Cancer of ICL. LANCET EAL project will consist in the development of a tight collaboration for a minimally invasive tissue identification approaches using ambient mass spectrometry. This EAL is deriving from a tight collaboration since 2014 with Pr. Z. Takats on novel guide surgery instrument. In fact, Pr Z. Takats has developed a MS based technology (Intelligent Knife or i’Knife system) allowing for real time monitoring during surgery by collecting aerosol (smokes) liberated during tissue excision with an electric scalpel (Balog et al., 2013). This pioneer work has demonstrated the high potential of MS for providing signature MS spectra of the various areas of the tissues with possible distinction of normal and cancerous parts of tissues and the intermediate parts corresponding to the excision margins (Takats et al., 2012). Dr. Takats has settled a first prototype installed in a surgery room at Saint Mary Hospital in London (Balog et al., 2010; Schäfer et al., 2011). For diagnosis the system operates a collection of the molecular profiles and comparison of the molecular profiles to reference profiles implemented into a database of the various stages and grades of one cancer pathology. In parallel, PRISM has developed a novel instrument for in vivo real time and non-invasive diagnosis, the SPIDERMASS. This instrument consists in an innovative sampling probe based on laser ablation coupled to analytical instrument to perform real-time analysis (Fatou et al., 2016, Patent FR1458925). The instrument will aim to be used by surgeons in the operating room so as to improve the decision making. The system is based on tissue sampling correlated with a real-time analysis. LANCET EAL is based on the development of new methods lies in their minimally invasive nature, which is achieved in two, markedly different ways. In case of solid tumours deeply embedded into healthy tissue environment, a special sampling device will be developed and used which extracts only the molecules of interest from the tissues without removing any cellular component. This is achieved by using the so-called Solid Phase Microextraction method for the rapid extraction of hydrophobic (mostly lipid-type) tissue constituents. A metal fibber coated with solid phase extraction medium is inserted into the tumour and the extraction medium is exposed to the tissue. The fibre is retracted and subsequently analysed by Desorption Electrospray Ionization Mass Spectrometry (DESI-MS) to provide longitudinally resolved information on the tissue environment. In case of luminal (e.g. ovarian cancers, sarcoma) tumours we are planning to develop a sampling solution that will be compatible with endoscopic instrumentation. Atmospheric Pressure Laser Ablation (APLA) based desorption/ionization is the technology of choice for tissue sampling in this case. APLA has various advantages including i) possible miniaturization through the use of fibres for both laser light guiding to the tissue and material collection, ii) low damage to patient’s tissues with optimized laser energy and wavelength and iii) possible use of the laser light to destroy the cells for treatment perspectives. This means that APLA using fibered laser should provide a good solution for including it in an endoscopic device. This also means that the system can potentially be combined with a surgical robot and be used for robotic surgery, even for brain pathologies such as Epilepsy. Through LANCET, PRISM and Pr. Z. Takats have started to publish a common paper in Cancer Cell, and have applied to ANR, but also in H2020.