Abstract
The brain has an exceptionally high requirement for energy metabolism, with glucose serving as the exclusive energy source. Cancers, including glioblastoma, have a high glucose uptake and rely on aerobic glycolysis for energy metabolism. The alternation of high-efficiency oxidative phosphorylation to a low-efficiency aerobic glycolysis pathway (Warburg effect) provides macromolecules for biosynthesis and proliferation. Current research indicates that the specific metabolism in the tumor tissue and normal brain tissue in the glioma allows the use of 5-aminolevulinic acid (5 ALA)-induced protoporphyrin IX (PpIX) and methylene blue (MB) to monitor and correct the development of the tumor. The focus is on the detection of the differences between tumor cells and tumor-associated macrophages/microglia using spectroscopic and microscopic methods, based on the fluorescent signals and the difference in the drug accumulation of photosensitizers (PSs). Since 5 ALA has long been used effectively in the clinic for fluorescent surgical navigation, it was employed as an agent to identify the localization of tumor tissue and study its composition, particularly tumor and immune cells (macrophages), which have also been shown to actively accumulate PpIX. However, since PpIX is photodynamically active, it can be considered effective as the main target of tumor tissue for further successful photodynamic therapy. MB was employed to visualize resident microglia, which is important for their activation/deactivation to prevent the reprogramming of the immune cells by the tumor. Thus, using two drugs, it is possible to prevent crosstalk between tumor cells and the immune cells of different geneses.
Keywords: fluorescent diagnostics, spectroscopic method, video fluorescent method, photosensitizer (PS), brain, microglia, macrophages, 5-aminolevulinic acid (5 ALA), methylene blue (MB)
Acknowledgements
The reported study was funded by Russian Foundation for Basic Research according to the research project No. 18-29-01062.
Footnotes
Yulia S. Maklygina received her M.Sc. degree (2013) in Physics from Lomonosov Moscow State University, Russia and her Ph.D. degree (2019) in Laser Physics from A.M. Prokhorov General Physics Institute, Russia. During her researching, she developed the new methods of spectroscopy and photophysics. She is interested in the absorption, fluorescence, FRET, dynamic light scattering, laser scanning confocal, FLIM, small animal imaging. The experimental models which she applied are: the tumoral culture in monolayer and 3D cell cultures (spheroids), growth, cytotoxicity, phototoxicity and apoptosis tests, uptake, intracellular drug localization. Now she is working as a researcher at Prokhorov General Physics Institute of the Russian Academy of Sciences, Russia. The area of scientific interests includes biophotonics and various methods of optical spectroscopy applied to the differentiation of biological tissues.
Alexei S. Skobeltsin received his M.Sc. degree (2019) in Medical Biophysics from Pirogov Russian National Research Medical University, Russia. Now (since 2020) he is a Ph.D. student at National Research Nuclear University MEPhI, Russia, and junior scientist in Laser Biospectroscopy Lab at Prokhorov General Physics Institute of the Russian Academy of Sciences, Russia. He is interested in bio-photonics, cancer biology, immunology, and confocal microscopy.
Tatiana A. Savelieva received her Master of Engineering degree with a specialization in Engineering in biomedical practice in 2005 and Ph.D. degree in Laser Physics in 2013 from A.M. Prokhorov General Physics Institute of the Russian Academy of Sciences, Russia.
Now Tatiana A. Savelieva is working as a researcher at General Physics Institute and an associate professor at National Research Nuclear University MEPhI, Russia. The area of scientific interests includes biophotonics and various methods of optical spectroscopy applied to the differentiation of biological tissues.
Galina V. Pavlova received her Ph.D. degree from Institute of Gene Biology, Russian Academy of Sciences, Russia in 2010. Presently, she is the Head of Laboratory of Neurogenetics and Developmental Biology at the same Institute in Russia. Also she is a professor at Pirogov Russian National Research Medical University (RNRMU). Her current research is focused on the study of neural differentiation potential of mammals stem and progenitor cells. She also explores proliferation and differentiation of cells in pathological conditions (for example, human glioblastoma).
Ivan V. Chekhonin received his M.Sc. degree (2016) in Medicine from Lomonosov Moscow State University, Russia. During his graduate studies, he developed this direction: “Pulsed dendritic cells in treatment of experimental glioma,” carried out in collaboration with V.P. Serbsky National Medical Research Centre of Psychiatry and Narcology, Russia. He received qualification in neurology from Lomonosov Moscow State University in 2018 (joint residency program with Research Center of Neurology, Moscow). He received qualification in radiology (diagnostic imaging) from A.I. Yevdokimov Moscow State University of Medicine and Dentistry in 2019 (professional retraining program). He is a Ph.D. student at N.N. Burdenko National Medical Research Center of Neurosurgery (Moscow, Russian) from 2018 to date (Ph.D. thesis: “Magnetic resonance relaxometry in brain glioma assessment”). He is interested in: magnetic resonance imaging, magnetic resonance relaxomerty, neuroradiology, neuro-oncology, neuroimmunology, glioma experimental modeling, and dendritic cells.
Olga I. Gurina received her M.Sc. degree (1990) in Pediatrics from Kuban State Medical University, Russia. She received her Ph.D. degree in Pediatrics and Biochemistry in 1996 (thesis: “Clinical and immunological assessment of blood-brain barrier function impairment in premature children with perinatal central nervous system injuries”). She received her D.Sci. degree in Biochemistry in 2005 (thesis: “Monoclonal antibodies to neurospecific proteins. Production, immunochemical analysis, study of blood-brain barrier permeability”). Now she is professor and corresponding member in Russian Academy of Sciences, Russia. She is interest in: studies of neurospecific protein spectrum in human and animals and its role in neurological and psychiatric disorders, monoclonal antibody- and neurospecific antigen-assisted assessment of blood-brain barrier function, role of blood-brain barrier in maintaining brain tissue homeostasis, and blood-brain barrier response to extreme stimuli.
Anastasiya A. Chernysheva received her M.Sc. degree (2017) in Biology with honors from Voronezh State University, Russia. Noe she is junior researcher at V.P. Serbsky National Medical Research Centre of Psychiatry and Narcology, Moscow, Russian from 2019 to date. She is interested in: biochemistry, immunology, and dendritic cell therapy in oncology.
Sergey A. Cherepanov received his M.Sc. degree (2013) in Medical Biophysics from Pirogov Russian National Research Medical University, Russia. He received his Ph. D. degree from Pirogov Russian National Research Medical University in 2019 (thesis: “Influence of Hedgehog signal pathway components on high-grade glioma proliferation and chemoresistance”). He has mastered methods, such as flow cytometry and cell sorting, PCR, immunochemistry, magnetic resonance imaging, and Ivis spectrum CT. Now he is junior researcher/Researcher at V.P. Serbsky National Medical Research Centre of Psychiatry and Narcology, Moscow, Russian from 2016 to date. He is interested in: radiology, magnetic resonance imaging and its physical basis, computed tomography, oncology, neurology, and chemoresistance.
Victor B. Loschenov received his M.Sc. degree (1976) in Electrical Engineering from Moscow Energy Institute, Russia. He received his Ph.D. degree (1981) in Quantum Radio Physics from N.S. Kurnakov Institute of General and Inorganic Chemistry, Academy of Sciences of the USSR. He received his Ph.D. degree (2006) in Laser Physics from Prokhorov General Physics Institute of the Russian Academy of Sciences, Russia. Since 2007, he has a title of professor in Laser physics in Prokhorov General Physics Institute of the Russian Academy of Sciences, Russia.
Now he is professor and head of laboratory at Prokhorov General Physics Institute of the Russian Academy of Sciences, professor at National Research Nuclear University (NRNU) MEPhI, Russia. He is interested in developing the following methods: photodynamic therapy and fluorescent diagnostics of cancer, medical devices, theranostics, and optical spectroscopy. His is the general director of OOO BIOSPEC (Moscow, Russia).
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