Heat Shock Protein Inducer Suppresses Osteogenic Differentiation of Human Mesenchymal Stem Cells

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Abstract

The key processes determining cell differentiation are the synthesis of new proteins and restructuring of the extracellular matrix. The production of matrix and cytosolic proteins, in turn, requires the constant involvement of the heat shock protein system — chaperones that ensure the proper folding of newly synthesized protein molecules and maintain their functional state. The aim of this study was to evaluate the effect of the heat shock protein inducer geranylgeranylacetone (GGA) on the differentiation of human mesenchymal stem cells. The study found that pretreating cells with GGA for 24 hours altered the pattern of calcium responses to the pro- osteogenic stimulus, parathyroid hormone. There was an increase in the number of cells responding to anti- osteogenic calcium oscillations associated with a decrease in the osteogenic potential of MSCs. Induction of osteogenic differentiation demonstrated a decrease in calcium deposition and RUNX2 and OSX gene expression. However, the addition of GGA had no significant effect on adipogenic differentiation. In search of a possible anti- osteogenic mechanism for the action of GGA, we analyzed the MSC proteomes treated with GGA or DMSO as controls. We found that incubation with GGA resulted in the absence of several proteins. Among these, calmodulin proteins (CALM1, CALM2, and CALM3), coactivators of adenylate cyclases, and proteins that regulate the actin cytoskeleton play a key role in osteogenesis. Thus, we demonstrated that the effect of GGA leads to a significant reduction in the capacity of MSCs for osteogenic, but not adipogenic, differentiation. A more detailed study of the mechanism of GGA action on embryonic and postnatal stem cells will further elucidate key features of the formation of new osteoblasts during embryogenesis and postnatal tissue renewal.

About the authors

V. A Usachev

Lomonosov Moscow State University

Email: usachjov-vova@mail.ru
ORCID iD: 0000-0003-1564-0644
Moscow, Russian Federation

M. V Shchebetina

Lomonosov Moscow State University

Email: mashaaasachcorg@gmail.com
ORCID iD: 0009-0006-1674-6247
Moscow, Russian Federation

K. Yu Smazhilo

Lomonosov Moscow State University

Email: k.smazhilo@yandex.ru
ORCID iD: 0009-0005-2349-4626
Moscow, Russian Federation

M. A Kulebyakina

Lomonosov Moscow State University

Email: m.a.kulebyakina@gmail.com
ORCID iD: 0000-0001-5956-1265
Moscow, Russian Federation

O. I Klychnikov

Lomonosov Moscow State University

Email: oklych@yahoo.co.uk
ORCID iD: 0000-0002-0383-9237
Moscow, Russian Federation

E. Bakhchinyan

Lomonosov Moscow State University

Email: is5492@mail.ru
ORCID iD: 0009-0000-9418-0396
Moscow, Russian Federation

N. S Voloshin

Lomonosov Moscow State University

Email: nik.voloshin.98@mail.ru
ORCID iD: 0009-0006-4209-3470
Moscow, Russian Federation

M. A Zamotina

Lomonosov Moscow State University

Email: zamotina.maria00@gmail.com
ORCID iD: 0000-0001-9994-6784
Moscow, Russian Federation

P. A Tyurin-Kuzmin

Lomonosov Moscow State University

Email: tyurinkuzminpa@my.msu.ru
ORCID iD: 0000-0002-1901-1637
Moscow, Russian Federation

M. V Vorontsova

Lomonosov Moscow State University

Email: maria.v.vorontsova@mail.ru
ORCID iD: 0000-0002-9124-294X
Moscow, Russian Federation

K. Yu Kulebyakin

Lomonosov Moscow State University

Email: konstantin-kuleb@mail.ru
ORCID iD: 0000-0001-6954-5787
Moscow, Russian Federation

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