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건국대학교

J. Adv. Res., Sep 28 2023, S2090-1232(23)00290-4 | https://doi.org/10.1016/j.jare.2023.09.041 Efficient improvement of the proliferation, differentiation, and anti-arthritic capacity of mesenchymal stem cells by simply culturing on the immobilized FGF2 derived peptide, 44-ERGVVSIKGV-53.

Soo Bin Lee ^a,1, Ahmed Abdal Dayem ^a,1, Sebastian Kmiecik ^b, Kyung Min Lim ^a, Dong Sik Seo ^c, Hyeong-Taek Kim ^c, Polash Kumar Biswas ^a, Minjae Do ^d, Deok-Ho Kim ^d, Ssang-Goo Cho ^a

^aDepartment of Stem Cell & Regenerative Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea

^bBiological and Chemical Research Centre, Faculty of Chemistry, University of Warsaw, 02-089 Warsaw, Poland

^cStem Cell Research Center of AMOLIFESCIENCE Co., Ltd, 91, Gimpo-daero 1950 beon-gil, Tongjin-eup, Gimpo-si, Gyeonggi-do 10014, Republic of Korea

^dDepartment of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21205 USA

¹Co-first authors.

Corresponding author : Ssang-Goo Cho

Abstract

The stem cell microenvironment has been evidenced to robustly affect its biological functions and clinical grade. Natural or synthetic growth factors, especially, are essential for modulating stem cell proliferation, metabolism, and differentiation via the interaction with specific extracellular receptors. Fibroblast growth factor-2 (FGF-2) possesses pleiotropic functions in various tissues and organs. It interacts with the FGF receptor (FGFR) and activates FGFR signaling pathways, which involve numerous biological functions, such as angiogenesis, wound healing, cell proliferation, and differentiation. Here, we aim to explore the molecular functions, mode of action, and therapeutic activity of yet undetermined function, FGF-2-derived peptide, FP2 (44-ERGVVSIKGV-53) in promoting the proliferation, differentiation, and therapeutic application of Wharton's jelly mesenchymal stem cells (WJ-MSCs) in comparison to other test peptides, canofin1 (FP1), hexafin2 (FP3), and canofin3 (FP4) with known functions. The immobilization of test peptides that are fused with mussel adhesive proteins (MAP) on the culture plate was carried out via EDC/NHS chemistry. Cell Proliferation assay, colony-forming unit, western blotting analysis, gene expression analysis, RNA-Seq. analysis, osteogenic, and chondrogenic differentiation capacity were applied to test the activity of the test peptides. We additionally utilized three-dimensional (3D) structural analysis and artificial intelligence (AI)-based AlphaFold2 and CABS-dock programs for receptor interaction prediction of the peptide receptor. We also verified the in vivo therapeutic capacity of FP2-cultured WJ-MSCs using an osteoarthritis mice model. Culture of WJ-MSC onto an FP2-immobilized culture plate showed a significant increase in cell proliferation (n = 3; *p < 0.05, **p < 0.01) and the colony-forming unit (n = 3; *p < 0.05, **p < 0.01) compared with the test peptides. FP2 showed a significantly upregulated phosphorylation of FRS2α and FGFR1 and activated the AKT and ERK signaling pathways (n = 3; *p < 0.05, **p < 0.01, ***p < 0.001). Interestingly, we detected efficient FP2 receptor binding that was predicted using AI-based tools. Treatment with an AKT inhibitor significantly abrogated the FP2-mediated enhancement of cell differentiation (n = 3; *p < 0.05, **p < 0.01, ***p < 0.001). Intra-articular injection of FP2-cultured MSCs significantly mitigated arthritis symptoms in an osteoarthritis mouse model, as shown through the functional tests (n = 10; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001), modulation of the expression level of the pro-inflammatory and anti-inflammatory genes, and improved osteochondral regeneration as demonstrated by tissue sections. Our study identified the FGF-2-derived peptide FP2 as a promising candidate peptide to improve the therapeutic potential of WJ-MSCs, especially in bone and cartilage regeneration.

논문정보

형식Research article
게재일2023년 09월 (BRIC 등록일 2023-10-05)
연구진국내(교신)+국외 연구진
분야 생명과학 > 분자생물학

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