Сell physiology and pathology laboratory

Mechanism of neurodegeneration: an ancient molecule as an essential element of brain cell physiology and pathology

Relevance of the project

Ensuring the functioning of the healthcare system is one of the priorities of the Russian Federation's state policy (according to forecasts, federal budget expenditures on healthcare in 2024 will amount to 1.62 trillion rubles (4.6% of the total planned expenditures). Within the framework of the national project “Health Care” the costs will amount to 289.9 billion rubles, and within the framework of the state programme “Health Care Development” in 2024-2026 – at least 3.99 trillion rubles). It should be noted that neurodegenerative diseases form a serious workload on other branches of the social sphere related to the support of people affected by these pathologies. In this regard, the research planned under the project, which will not only provide new fundamental knowledge about the mechanisms of neurodegeneration, but will also develop the understanding of natural mechanisms of neuroprotection, as well as discover promising pharmacological targets, may form the basis for the development and introduction of domestic drugs based on physiologically active substances for the correction of the state or full restoration of brain functions of patients with neurodegenerative diseases, the majority of which will be used in the development and implementation of new drugs for the treatment of neurodegenerative diseases.

The intended research corresponds to the priority areas of technological sovereignty projects and projects for structural adaptation of the economy of the Russian Federation, approved by Resolution of the Government of the Russian Federation No. 603 of 15.04.2023, namely the “medical industry” area in terms of the production of materials, bandages and similar products, including those coated with medications. The use of the unique potential of polyphosphate can provide the creation of materials with high efficiency of tissue regeneration, reducing both recovery time and suffering of patients, as well as avoiding negative side effects.

Aim of the project

Studying the pathogenesis and mechanism of neuronal death in major neurodegenerative diseases, the role of inorganic polyphosphate in neuronal and astrocytic signaling, identifying the involvement of polyphosphate in the molecular and cellular mechanisms of pathology in neurodegenerative diseases, and investigating its modulation potential and applications in pharmaceuticals and bioengineering.

Project Goals

2024

1. Determination of the causes of changes in polyphosphate levels in hereditary forms as well as in toxic models of neurodegenerative diseases.
2. Development of a method to assess the degree of contribution of flavinadenine dinucleotide (FAD++), which is a part of various flavoproteins, to the total pool of FAD++ autofluorescence in vitro (on intact mitochondria, healthy fibroblasts, neuron-glial mixed culture and on cell cultures - models of neurodegenerative diseases) using mitochondrial substrates, inhibitors of mitochondrial electron-transport chain complexes, as well as inhibitors of flavin enzymes (in particular, MAO).

2025

1. Detection, localization and quantitative analysis of specific enzymes providing ATP synthesis at the expense of polyphosphate.
2. Investigation of activation conditions as well as features of enzyme kinetics of polyphosphate-dependent ATP synthesis at the level of isolated enzymes, intact mitochondria, and in mitochondria of permeabilized and intact cells.
3. Development of a method to assess the degree of contribution of FAD++, which is part of various flavoproteins, to the total pool of autofluorescence of FAD++ in the study of acute brain slices of healthy animals and animals of toxic models of neurodegenerative diseases (in particular, Parkinson's disease) using mitochondrial substrates, inhibitors of mitochondrial electron-transport chain complexes, as well as inhibitors of flavin enzymes (in particular, MAO).

2026

1. Study of the effects of extracellular and intracellular polyphosphate on dopamine, glutamate and other neurotransmitter-stimulated signaling in control cells and cellular models of neurodegenerative diseases.
2. Investigation of the role of polyphosphate in mPTP opening in cellular models of neurodegenerative diseases.
3. Development of a method for differentiation of mitochondrial FAD++ signal between possible sources when performing in vivo studies without the use of substrates and inhibitors.

2027

1. Determination of a potential interaction between polyphosphate and α-synuclein, β-amyloid or tau protein in neurodegenerative diseases.
2. Establishing a possible protective effect of modulating polyphosphate levels against neurodegeneration in neurodegenerative diseases.

2028

1. Study of the role of polyphosphate-dependent ATP synthesis in energy-consuming processes and under hypoxia/anoxia conditions.
2. Study of the prospects for the use of polyphosphate in wound-healing products and as a base or component of composite material for 3D printing or 3D bioprinting in solving the problems of bioengineering of tissues and organs.

Expected results of the project implementation

Scientific results of the project

The ongoing project will answer several fundamental scientific questions. The planned research will help identify the intracellular localization of polyphosphate in healthy neurons and astrocytes, and reveal how this compartmentalization changes in neurodegenerative diseases. Inorganic polyphosphate is a gliotransmitter, but its effects on neurons are still underestimated. Therefore, the results of this project will help to define the role of polyphosphate in neuronal signal transduction and provide insight into how polyphosphate interacts with signals induced by other neurotransmitters, particularly glutamate and dopamine. All of these basic questions are important for understanding the cellular mechanisms of neurodegeneration. The role of inorganic polyphosphate in the aggregation of specific proteins (particularly α-synuclein) that play an important role in PD and other neurodegenerative disorders will also be assessed. The results of this project will help to clarify whether changes in intracellular polyphosphate concentration can protect the brain from neuronal cell loss in pathologies.

Studies planned within the project to identify regulators of polyphosphate levels (both those involved in the synthesis and those consuming the polymer) in brain cells will help to identify therapeutic targets, the impact on which by using physiologically active substances can provide a level of neuroprotection sufficient to prevent, slow down or completely stop certain neurodegenerative processes.

In addition, the project will develop hypotheses on the prospects for the application of polyphosphate in pharmaceuticals and bioengineering. In particular, research will be carried out in the field of creation of polyphosphate-containing bandages, the increased efficiency of which may be due to the provision of regenerative processes with energy released by hydrolysis of phosphorus-anhydride bonds in the polymer structure. The identified optimal parameters (degree of polyphosphate polymerization, type of counterion, phase state of polymer particles) as well as the technology of bandages manufacturing will be protected by patents of the Russian Federation for invention. The project will also investigate the prospects of using polyphosphate as a material (as a base material or in combination with other high molecular weight compounds) for 3D printing of matrices for tissue engineering or 3D bioprinting. Through gradual biodegradation, polyphosphate will allow the creation of a temporary base for implantation that will provide a high rate of cell proliferation and differentiation followed by complete replacement with fully functioning tissue.

Development of the material and technical base of the laboratory and professional development of the staff

Within the framework of the project implementation, it is planned to solve a number of organisational and infrastructural tasks related to the involvement of talented young people in the implementation of advanced research, creation of conditions for scientific work, including the development of the existing material and technical base. With the intention of significant development in the direction of providing in vivo work in accordance with the existing norms of the Russian legislation, as well as approaches applied in the world research practice, it is planned to create a block of rooms for temporary housing of small laboratory animals that do not belong to the category of free from specific microflora (SPF category). To accommodate microscopic, analytical, imaging and other laboratory equipment, additional rooms will be allocated and prepared to meet all necessary safety, arrangement and operation rules.

The following scientific equipment is planned to be purchased to fulfil the set objectives:

- time-resolved confocal fluorescence microscope;

- a set of equipment for isolating individual cells and analyzing the transcription level of target genes by quantitative PCR;

- multimodal spectral OCT unit of the second generation;

- a system for laser speckle contrast and hyperspectral imaging;

- ultrasound high-frequency Doppler;

- equipment for 3D printing of structures for culturing cells on their surface and 3D bioprinting using modified bioinks.

The project also provides for additional professional education of the project team members in the form of internships and training programmes and attendance of methodological seminars. These activities are aimed at the formation of new professional competences in order to ensure that the project team's qualifications meet the changing conditions of professional activity.

We always welcome offers of mutually beneficial collaboration!