Immunoglobulin G as an innovative neuro-diagnostic agent
In a number of neurodegenerative diseases (NDs) the production of the anti-neuronal immunoglobulin G (IgG) is a significant feature of the immune-inflammatory process. It was shown that human IgGs may induce diverse physiological effects not only on neurons but also on glial cells of animal origin. Of note, these reactions were only in a few cases explained as an antibody-antigen reaction. We propose a genuinely novel biomedical approach to the use of patient IgGs to induce and study physiological reactions on a standardized cellular test platform for in vitro screening of IgGs from amyotrophic lateral sclerosis (ALS) patients.
As a short-term goal, we would like to explore the mechanism(s) of these non-canonical IgG effects. As a long-term goal, we intend to introduce a disruptive molecular diagnostics technology at the cellular imaging level that may allow better therapy follow-up of ALS. This unique approach offers a multipurpose processing of a single sample (patient’s IgGs) activating physiological response markers on cells cultured in a microfluidic lab-on chip. The chip can provide complex information about multiple cellular signaling processes recorded by: (1) electrophysiology – voltage imaging, (2) calcium imaging, (3) Reactive Oxygen Species (ROS), and (4) intracellular vesicle trafficking.
This experimental approach will lead to a clinically relevant technology based on microfluidics and automated on-chip video-microscopy. The multidisciplinary data analysis methodology with machine learning will improve the performance towards diagnosis, prediction, monitoring, intervention and especially the assessment of therapeutic response. The personalized experimental diagnostics designed for ALS should in long term serve a variety of NDs in the particular area of neuroinflammation.
List of work packages (WP)
- Standardization and benchmarking of transduction mechanisms of NDs IgG effects
- rigorous standardization of experimental protocols
- selection of best practice
- benchmarking of protocols
- Design of the optical system and recording software
- designing the integrated optics-on-chip
- defining quantitative parameters with machine learning protocols
- conversion of activity parameters into classification accuracy
- Pilot platform design
- design and validation of the lab-on-a-chip platform
- flow simulation and testing of the microfluidics device
- pressure control in the microfluidics device
- development of an integrated data acquisition system
- testing of cell culturing and maintenance in the lab-on-a-chip
- Management and sustainability
Credentials of PI and members of Project team
Pavle R. Andjus (male, WP3 Coordinator) – Principal investigator, full professor, head of Center for laser microscopy Faculty of Biology University of Belgrade. Coordinator of the first FP project in biomedicine in Serbia – FP6 project “NEUROIMAGE”, also coordinated a trilateral ERA NET project “NanoHyperRadicals” and 5 bilateral projects (Croatia, France, Hungary, Ukraine, Italy), three DAAD projects with Germany and two large national projects (until recently a large Integrative, interdisciplinary research project “Biomarkers in neurological and malignant processes” – MESTD Republic of Serbia grant #41005), until current year coordinated also an innovative Proof of concept grant by the Innovation Fund of Republic of Serbia. Current project activities: Coordinates a H2020 MSCA RISE project “AUTOIGG” (with 9 partners from USA – 3, Costa Rica – 1, Turkey – 2, France – 1, Finland – 1 and Serbia – 1), and one bilateral DAAD grant and participates in another one. He has pursued successfully, towards the finals, several innovation competitions with his team – IMAGINE IF (UK), Brain Innovation Days (Brussels), and currently Jumstarter – EIT and 100 BestTech (China). He is president of the Serbian brain Council (representative of the National Councils in the European Brain Council), Council member of the European Society for Neurochemistry, Executive committee member of the Biophysical Society of Serbia, Member of University Council for Multidisciplinary studies and president of its Ethical committee.
Lidija Radenović (female, WP4 Coordinator) – full professor, Center for laser microscopy Faculty of Biology University of Belgrade Research interests: Neuroscience – Brain trauma; brain homeostasis; neurodegenerative diseases; ALS; stroke; neuroprotection; nervous system plasticity; neurogenesis; neurobiology of behavior, neuroimaging, mitochondria, oxidative stress, neuroinflammation, blood-brain barrier, biomarkers, neuroinflammation, therapy… Participated in 8 national and 3 international projects. Winner of Weiser Professional Development Award for Study and Research at the University of Michigan, USA in 2012; 2018 and 2019 served as visiting scientist at SME ArGenit, Medipol University, and Yeditepe University, Istanbul, Turkey, financed by the AUTOIGG MSCA RISE project. 2020. Visiting scientist at LANOTEC – Laboratorio national de nanotechologia, National High Technology Center, San Jose, Costa Rica, financed by the AUTOIGG project.
Andrej Korenić (male, WP2 Coordinator) — holding a Ph.D. in neurobiology, has been a valuable team member in previous projects, responsible for advancing our understanding of cerebral ischemia through both in vitro and in vivo studies. With a solid foundation in neurobiology/neuroscience, cellular physiology & biophysics, electrophysiology and computational neuroscience, his research extends to collaborative projects on amyotrophic lateral sclerosis (ALS), enhancing these studies with statistical analyses and automation of analyses routines. His recent endeavors, since 2019, focus on integrating AI tools with neuroscience and biomedical data analysis, showcasing a forward-thinking approach to utilizing machine learning in aiding the field of biomedicine to obtain more detailed, faster, and more accurate analyses.
Katarina Živančević (female) – PhD student at the University of Belgrade – Faculty of Pharmacy, as well as Research Trainee at the Faculty of Biology – Center for laser microscopy. She is engaged in the market analyses and business development. Her main role in NIMOCHIP development is research and marketing strategy, business development as well as presentation of the product. Pitched this technology at several innovation fairs and competitions.
Ana Jakovljević (female) – Research Assistant at the Center for Laser Microscopy, Faculty of Biology, University of Belgrade. PhD student of Biophotonics, at Interdisciplinary studies at University of Belgrade; 2017 MSc, Biophysics, Faculty of Biology, University of Belgrade; 2022 seconded to the Medipol University, Istanbul, Turkey with the H2020 AUTOIGG project; 2021 Short scientific visit to the Department of Neuroanatomy, Medical Faculty, University of Bochum, Germany (bilateral DAAD project “Neurorepair by proteolysis of the cell adhesion molecule L1CAM in ALS and spinal cord injury models”) 2019 study visit Institute for Molecular Genetics, Light Microscopy Core Facility, Prague, Czech Republic. Honours: 2019, Art of Neuroscience, Netherlands Institute for Neuroscience.
Zorica Stević (female) – Professor of Neurology, Institute of neurology, Faculty of Medicine, University of Belgrade. More than 40 publications in distinguished international medical journals in area of neuromuscular and neurodegenerative disorders. Special research interest in amyotrophic lateral sclerosis and motor neuron diseases. Member of the European ALS Consortium, European FALS Consortium, EURALS Consortium and the Member of the Board of Directors of the International Alliance of ALS/MND Associations. She will serve as a consultant and source for the selection for patient samples. Author of 76 publications with citations 1367, and H-index 21. Collaborates with PI and the CLM team since the 90’s.Role in NIMOCHIP will be to select and characterize patients for sera and IgG isolation.
Irena Živković (female) – Principal Research Fellow, Institute of Virology, Vaccines and Sera “Torlak”. More than 50 publications in distinguished international immunology journals in the area of humoral immunology, with emphasis on natural and induced antibodies. Special research interest in autoimmune diseases (ALS, APS) and their animal models, as well as the influence of eugenic factors (immunization) on the regulatory network of natural antibodies. Her main role in NIMOCHIP development is research on antibody properties in ALS patients and their role in disease induction.