ИСТИНА

Целью проекта является выяснение роли миРНК, секретируемых МСК человека в составе внеклеточных везикул, в реверсии фиброза. Задачи проекта: 1) Проанализировать набор миРНК, секретируемых МСК человека в составе внеклеточных везикул, для выявления ассоциированных с процессами подавления и обратного развития фиброза миРНК. 2) Получить репортерные линии клеток, экспрессирующие белки-маркёры миофибробластов. 3) Отработать 3D модель мультиклеточной конструкции, имитирующей “фибротический фокус”. 4) Изучить влияние выбранных миРНК, секретируемых МСК человека в составе внеклеточных везикул, на реверсию трансдифференцировки миофибробластов на 2D и 3D моделях фиброза in vitro.

Fibrosis is one of the possible pathological consequences of the regeneration program violation caused by chronic damaging factors. The key markers of fibrogenesis are an increase the pool of myofibroblasts and their excessive synthesis protein of extracellular matrix (ECM), that leads to disruption of the structure and function of tissues and organs and the development of severe fibroproliferative diseases with an unfavorable prognosis. The development of chronic fibrosis is also accompanied by an interruption of the regulatory function of the immune system and the inability of the microenvironment to regulate dedifferentiation and elimination of myofibroblasts. However, the interaction of inductors and regulators of these disorders has been studied insufficiently, therefore, the study of effective tools for suppressing or reversing fibrosis is the most relevant. Mesenchymal stromal cells (MSCs) are believed to be key regulators capable of inhibiting progression of fibrosis in various tissues. Today, regulatory microRNAs (miRNAs) secreted by MSC within extracellular vesicles are considered to be one of the main mediators of the antifibrotic activity of MSCs. Previously, we demonstated the ability of extracellular vesicles secreted by MSCs to inhibit fibroblast differentiation induced by a key pro-fibrogenic stimulus, the transforming growth factor beta (TGFb), by direct transfer of miRNAs such as miRNA -21 and -29c to these cells. It is important to note that the only extracellular vesicles were part from MSC secretome that also were able to stimulate the dedifferentiation of myofibroblasts, and due to this effects may influence the restoration of the normal tissue structure. However, the precise mechanisms of this phenomenon remain unexplored. Therefore, in the work, it is planned to establish the effect of miRNAs within extracellular vesicles secreted by human MSCs on the reversal of fibrosis in 2D and 3D cell models. Bioinformatic sсan will be held for the search main miRNAs contained in the extracellular vesicles of human MSCs and involved in the dedifferentiation of myofibroblasts based on the data of our previous next generation RNA sequencing and PCR screening. The contribution of the selected miRNAs will be established on the models of dedifferentiation of myofibroblasts using a wide range of cell, biochemiсal and molecular biology methods. As a key model for the study, it is planned to create a multicellular analogue of “fibrotic foci” as the main functional unit for the progression of pathological fibrosis. In this model, it is proposed to study the mechanisms of participation of extracellular vesicles secreted by MSCs and selected miRNAs in the regulation of transdifferentiation of myofibroblasts and remodeling of tissue-like fibrous constructs in real time. As a result of this project, new data about mechanisms of direct participation of extracellular vesicles secreted by MSCs and their miRNAs in the reversal of fibrosis will be obtained. The results can be the basis for further studies of the role of MSCs in the area of ​​pathological fibrosis in vivo, as well as for the development of new approaches to the treatment of severe fibrotic diseases using methods of regenerative medicine and “cell-free therapy”.