Project

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Assessment of the metabolic activity of biological tissues in-vivo by time-resolved luminescence imaging

English title Assessment of the metabolic activity of biological tissues in-vivo by time-resolved luminescence imaging
Applicant Wagnières Georges
Number 147141
Funding scheme Project funding (Div. I-III)
Research institution Institut des sciences et ingénierie chimiques EPFL - SB - ISIC
Institution of higher education EPF Lausanne - EPFL
Main discipline Other disciplines of Physics
Start/End 01.05.2013 - 30.04.2015
Approved amount 185'250.00
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All Disciplines (4)

Discipline
Other disciplines of Physics
Pathophysiology
Physiology : other topics
Biophysics

Keywords (9)

Oxygenation; Protoporphyrin IX; Biological tissues; In vivo; Lesion demarcation; Time-resolved luminescence imaging; Molecular Probes; Metabolism; Martorell's ulcer

Lay Summary (French)

Lead
La mesure de la teneur tissulaire en oxygène ainsi que du rôle qu'il joue dans le métabolisme sont d'une grande importance, que cela soit pour d'un point de vue fondamental ou appliqué. Malheureusement, il n'existe pas, à l'heure actuelle, de méthodes pratiques permettant de procéder à de telles mesures, en particulier en imagerie, in vivo ou en clinique.
Lay summary

Contenu et objectifs du travail de recherche

L'objectif général de ce projet consiste à développer de nouvelles méthodes permettant de visualiser l'oxygénation et l'activité métabolique de tissus biologiques basées sur l'imagerie et la spectroscopie résolues en temps de sondes moléculaires exogènes.

Un dispositif d'imagerie de temps de vie de luminescence sera développé afin d'effectuer des mesures à une échelle microscopique (études in vitro ou précliniques sur des équivalents épidermiques ou sur la membrane chorio-allantoïdienne de l'oeuf de poule) ou macroscopique (études précliniques ou cliniques en dermatologie). Selon le type de sonde moléculaire étudiée, ce dispositif sera utilisé pour générer des images de temps de vie de phosphorescence ou de fluorescence retardée. Pour cette deuxième application, ce dispositif d'imagerie sera obturable, cette caractéristique étant nécessaire pour rejeter la fluorescence prompte.

Contexte scientifique et social du projet de recherche

Les résultats permettront d'obtenir des informations utiles sur les mécanismes métaboliques intervenant dans l'apport et la consommation d'oxygène dans les tissus, en particulier lors de traitement pas thérapie photodynamique. De plus, ce projet fournira des informations fondamentales, utiles à la compréhension des mécanismes métaboliques jouant un rôle dans la production endogène de la protoporphyrine IX, un photosensibilisateur approuvé pour des applications cliniques.

Les applications médicales les plus importantes de l'approche proposée dans ce projet permettront de procéder à une démarcation, à un monitorage ainsi qu'à une individualisation du traitement de pathologies affectant la peau, tel que c'est le cas pour la kératose actinique ou l'ulcère de Martorell.

Direct link to Lay Summary Last update: 22.05.2013

Responsible applicant and co-applicants

Employees

Name Institute

Publications

Publication
Effect of PpIX photoproducts formation on pO2 measurement by time-resolved delayed fluorescence spectroscopy of PpIX in solution and in vivo.
Huntosova Veronika, Gerelli Emmanuel, Zellweger Matthieu, Wagnières Georges (2016), Effect of PpIX photoproducts formation on pO2 measurement by time-resolved delayed fluorescence spectroscopy of PpIX in solution and in vivo., in Journal of photochemistry and photobiology. B, Biology, 164, 49-56.
Fluence plays a critical role on the subsequent distribution of chemotherapy and tumor growth delay in murine mesothelioma xenografts pre-treated by photodynamic therapy.
Wang Yabo, Wang Xingyu, Le Bitoux Marie-Aude, Wagnieres Georges, Vandenbergh Hubert, Gonzalez Michel, Ris Hans-Beat, Perentes Jean Y, Krueger Thorsten (2015), Fluence plays a critical role on the subsequent distribution of chemotherapy and tumor growth delay in murine mesothelioma xenografts pre-treated by photodynamic therapy., in Lasers in surgery and medicine, 47(4), 323-30.
Optical properties of rabbit brain in the red and near-infrared: changes observed under in vivo, postmortem, frozen, and formalin-fixated conditions.
Pitzschke Andreas, Lovisa Blaise, Seydoux Olivier, Haenggi Matthias, Oertel Markus F, Zellweger Matthieu, Tardy Yanik, Wagnières Georges (2015), Optical properties of rabbit brain in the red and near-infrared: changes observed under in vivo, postmortem, frozen, and formalin-fixated conditions., in Journal of biomedical optics, 20(2), 25006-25006.
Endosomes: guardians against [Ru(Phen)(3)](2+) photo-action in endothelial cells during in vivo pO(2) detection?
Huntosova Veronika, Stroffekova Katarina, Wagnieres Georges, Novotova Marta, Nichtova Zuzana, Miskovsky Pavol (2014), Endosomes: guardians against [Ru(Phen)(3)](2+) photo-action in endothelial cells during in vivo pO(2) detection?, in METALLOMICS, 6(12), 2279-2289.
In vivo measurement of tissue oxygenation by time-resolved luminescence spectroscopy: advantageous properties of dichlorotris(1, 10-phenanthroline)-ruthenium(II) hydrate
Huntosova Veronika, Gay Sandrine, Nowak-Sliwinska Patrycja, Rajendran Senthil Kumar, Zellweger Matthieu, van den Bergh Hubert, Wagnieres Georges (2014), In vivo measurement of tissue oxygenation by time-resolved luminescence spectroscopy: advantageous properties of dichlorotris(1, 10-phenanthroline)-ruthenium(II) hydrate, in JOURNAL OF BIOMEDICAL OPTICS, 19(7), 077004-1-077004-12.
Low-Dose Vascular Photodynamic Therapy Decreases Tumor Interstitial Fluid Pressure, which Promotes Liposomal Doxorubicin Distribution in a Murine Sarcoma Metastasis Model.
Perentes Jean Yannis, Wang Yabo, Wang Xingyu, Abdelnour Etienne, Gonzalez Michel, Decosterd Laurent, Wagnieres Georges, van den Bergh Hubert, Peters Solange, Ris Hans-Beat, Krueger Thorsten (2014), Low-Dose Vascular Photodynamic Therapy Decreases Tumor Interstitial Fluid Pressure, which Promotes Liposomal Doxorubicin Distribution in a Murine Sarcoma Metastasis Model., in Translational oncology, 7(3), 393-399.
Temperature and oxygen-concentration dependence of singlet oxygen production by RuPhen as induced by quasi-continuous excitation
Varchola Jaroslav, Huntosova Veronika, Jancura Daniel, Wagnieres Georges, Miskovsky Pavol, Bano Gregor (2014), Temperature and oxygen-concentration dependence of singlet oxygen production by RuPhen as induced by quasi-continuous excitation, in PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES, 13(12), 1781-1787.
Correlation between protoporphyrin IX fluorescence intensity, photobleaching, pain and clinical outcome of actinic keratosis treated by photodynamic therapy.
Piffaretti Filippo, Zellweger Matthieu, Kasraee Behrooz, Barge Jérôme, Salomon Denis, van den Bergh Hubert, Wagnières Georges (2013), Correlation between protoporphyrin IX fluorescence intensity, photobleaching, pain and clinical outcome of actinic keratosis treated by photodynamic therapy., in Dermatology (Basel, Switzerland), 227(3), 214-25.
Measurement of pO2 by luminescence lifetime spectroscopy: A comparative study of the phototoxicity and sensitivity of [Ru(Phen)3 ](2+) and PdTCPP in vivo.
Huntosova Veronika, Gerelli Emmanuel, Horvath Denis, Wagnieres Georges, Measurement of pO2 by luminescence lifetime spectroscopy: A comparative study of the phototoxicity and sensitivity of [Ru(Phen)3 ](2+) and PdTCPP in vivo., in Journal of biophotonics.

Collaboration

Group / person Country
Types of collaboration
Prof. P. Miskovsky, Department of Biophysics, University Pavol Jozef Šafárik, Kosice Slovakia (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Forschungkolloquium KFSP Tumor Oxygenation Talk given at a conference Measurement of the tissue oxygenation by luminescence lifetime spectroscopy 17.09.2015 Zurich, Switzerland Wagnières Georges;
10th European Biophysics Congress Poster Albumin-[Ru(Phen)3]2+ complex formation: the influence of albumin on oxygenation measurement in vivo 18.07.2015 Dresden, Germany Wagnières Georges;
European Conference on Biomedical Optics 2015 Talk given at a conference Optical properties of the deep brain in the red and NIR: Changes observed under in vivo, post-mortem, frozen and formalin-fixed conditions 21.06.2015 Münich, Germany Wagnières Georges;
16th International Congress on Photobiology Poster Effect of ROS created during pO2 detection by [Ru(Phen)3]2+ luminescence lifetime measurements 08.09.2014 Cordoba, Argentina Wagnières Georges;
International Conference on Laser Applications in Life Sciences Talk given at a conference Tissue oxygenation by time-resolved luminescence spectroscopy 29.06.2014 Ulm, Germany Wagnières Georges;
Biophysical conference RBC Poster Tissular pO2 measurements by time-resolved luminescence spectroscopy: Correlation of results obtained in vivo with Ru(phen) and protoporphyrin IX 15.05.2014 Smolenice, Slovakia Wagnières Georges; Gerelli Emmanuel;
Annual meeting of the Biomedical Photonics Network Poster In vivo and in vitro measurement of the PPIX delayed fluorescence temperature dependence 01.11.2013 Berne, Switzerland Wagnières Georges; Gerelli Emmanuel;
Fall meeting 2013 of the Swiss Chemical Society Poster Measurement of the tissue oxygenation in vivo by time-resolved lumi-nescence spectroscopy of Ru(Phen), a poorly photosensitizing probe 06.09.2013 Lausanne, Switzerland Wagnières Georges;


Knowledge transfer events

Active participation

Title Type of contribution Date Place Persons involved


Communication with the public

Communication Title Media Place Year
Talks/events/exhibitions Courses entitled: Biomedical Optics and Photomedicine Western Switzerland 2013

Awards

Title Year
Best poster award, Regional Biophysics Conference, May 2014, Smolenice, Slovakia. 2014

Associated projects

Number Title Start Funding scheme
103518 Modulation and measurement of the oxygen concentration by reflectance and time-resolved luminescence spectroscopies to optimize the treatment of age-related macular degeneration 01.04.2004 Project funding (Div. I-III)
130518 Development of novel therapies based on the use of protoporphyrin IX precursors 01.05.2010 Project funding (Div. I-III)
130518 Development of novel therapies based on the use of protoporphyrin IX precursors 01.05.2010 Project funding (Div. I-III)
159746 Study of photobiostimulation by time-resolved and steady-state spectroscopy of protoporphyrin IX 01.04.2015 Interdisciplinary projects

Abstract

The general objective of this project is to develop new techniques to produce images of the oxygenation and metabolic activity of biological tissues by time-resolved luminescence spectroscopy of endogenous and exogenous molecular probes. This approach will provide useful fundamental information regarding the metabolic mechanisms that are in relation with the oxygen delivery and consumption, in particular during photodynamic therapy (PDT). In addition, this project will provide useful fundamental information regarding the metabolic activities involved in the endogenous production of a heme precursor, protoporphyrin IX (PPIX), an approved luminescent photosensitizer. Medical applications of the approach proposed here will enable to delineate, monitor and individualize therapies of conditions affecting the skin, such as actinic keratosis (AK) and Martorell's ulcer.More precisely, our first specific objective is to develop formulations and derivations of molecular probes that are sensitive to the oxygen partial pressure (pO2) in such a way that no photodamages are induced in the tissues during the measurement. These damages are observed with virtually all pO2 probes used in approaches based on the measurement of the oxygen quenching of their luminescence. Such “minimally invasive” pO2 probes will find numerous in vitro, preclinical and possibly clinical applications to achieve the general objective mentioned above.A second specific objective of this project is the individualization of the light dose during PDT of AK using the methylaminolevulinate (MAL)-induced PPIX delayed fluorescence. The new approach we propose in this project consists to exploit the quenching of the delayed fluorescence of an approved photosensitizer, the MAL-induced PPIX, to determine the oxygen concentration in the tissues and to adapt the therapeutic light dose accordingly. One major advantage of the approach proposed here, as compared to the “standard” approaches based on the use of the oxygen quenching of the phosphorescence of metallo-porphyrins, is related to its potential clinical use since we plan to use an approved photosensitizer for this purpose. This PPIX will also be used to image the metabolic activity and the tissue oxygenation in, and around, Martorell's ulcer to improve their demarcation. The main goal of this approach is to provide a guidance to improve the surgical resection of these lesions.Two time-resolved imaging systems will be developed to perform luminescence lifetime measurements on microscopic (in vitro or preclinical studies) or macroscopic (preclinical or clinical studies) samples. Different magnifications will be achieved by coupling these systems to dedicated optics (fluorescence microscope or "macro" objective). The imaging systems will be dedicated to applications where a spatial resolution of the pO2 measurement is of importance, whereas a point measurement system, already available in our laboratory, will be used when the luminescence of the molecular probe is too low to be imaged with a reasonable signal/noise ratio. The two imaging time-domain systems will be developed in such a way that their detector will be gatable. This optional feature will be used to reject the prompt fluorescence while measuring the lifetime of the PPIX delayed fluorescence. This feature is important since the delayed fluorescence presents itself as another component of fluorescence besides the prompt fluorescence, but having a decay time equal to the lifetime of the triplet state of the pO2 molecular probe.In parallel to these instrumental developments, we plan to synthesize several pO2 molecular probes presenting very poorly toxic and phototoxic properties as mentioned above. This activity will be based on our extensive know-how for the screening and characterization of photosensitizes, as well as on our know-how in the filed of the production of nanoparticles-based formulations. The strategy we plan to adopt for this subproject will be to select nanoparticles materials such that molecular oxygen can easily diffuse in this material, whereas the singlet oxygen produced by the quenching process is “neutralized” within the nanoparticle, thus preventing the generation of photodamages to the tissues.We will test these pO2 measurements technique using "advanced" tissue models. More precisely, the preclinical studies will be performed with our chick’s chorioallantoic membrane (CAM) model and with an epidermal equivalent model. These two different models were chosen since the first one enables to study the toxicity as well as the vascular effects of photosensitizers, whereas the second is more adapted to study these effects at a cellular or tissular level.Following these preclinical studies, it is anticipated that the first clinical applications will be for the delineation of Martorell's ulcers using the MAL-induced protoporphyrin IX delayed fluorescence. This clinical translation will be based on the long-term and fruitful collaboration established between our group and the Dermatology Unit of University Hospitals.It should be noted that the clinical availability of an imaging system to monitor the tissular pO2 using poorly phototoxic molecular probes or the PPIX delayed fluorescence will also pave the way to many new diagnostic and therapeutic applications expected by the medical community.Finally, the innovative approaches we propose above to image the tissular and intracellular oxygen concentration will lead to new insights into the transport and consumption of oxygen in biological tissues, as well as regarding our understanding of the mechanisms taking place during various therapies, including PDT. These insights will be novel since the intracellular location of photosensitizers resides at sites pivotal in the transport and utilization of oxygen (mitochondria). Therefore, our results and novel techniques will probably open new research area in related scientific and medical fields.
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