Research Paper|Volume 16, Issue 10|pp 8965—8979

CRYAB suppresses ferroptosis and promotes osteogenic differentiation of human bone marrow stem cells via binding and stabilizing FTH1

Bo Tian^1,⁵, Xiaolu Li^2,⁵, Weiyuan Li^2,⁵, Zhizhou Shi³, Xu He^2,⁵, Shengyu Wang³, Xun Zhu³, Na Shi³, Yan Li^2,⁵, Ping Wan^2,⁵, Chongtao Zhu^4,⁵

¹Scientific Research Section, The First People’s Hospital of Yunnan Province, Kunming 650032, China
²Geriatric Department, The First People’s Hospital of Yunnan Province, Kunming 650032, China
³Medical School, Kunming University of Science and Technology, Kunming 650500, China
⁴Laser Medical Center, The First People’s Hospital of Yunnan Province, Kunming 650032, China
⁵The Affiliated Hospital of Kunming University of Science and Technology, Kunming 650032, China

* Co first author

Received: October 24, 2023Accepted: March 25, 2024Published: May 22, 2024

https://doi.org/10.18632/aging.205851

Copyright: © 2024 Tian et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Background: Bone formation and homeostasis are greatly dependent on the osteogenic differentiation of human bone marrow stem cells (BMSCs). Therefore, revealing the mechanisms underlying osteogenic differentiation of BMSCs will provide new candidate therapeutic targets for osteoporosis.

Methods: The osteogenic differentiation of BMSCs was measured by analyzing ALP activity and expression levels of osteogenic markers. Cellular Fe and ROS levels and cell viability were applied to evaluate the ferroptosis of BMSCs. qRT-PCR, Western blotting, and co-immunoprecipitation assays were harnessed to study the molecular mechanism.

Results: The mRNA level of CRYAB was decreased in the plasma of osteoporosis patients. Overexpression of CRYAB increased the expression of osteogenic markers including OCN, OPN, RUNX2, and COLI, and also augmented the ALP activity in BMSCs, on the contrary, knockdown of CRYAB had opposite effects. IP-MS technology identified CRYAB-interacted proteins and further found that CRYAB interacted with ferritin heavy chain 1 (FTH1) and maintained the stability of FTH1 via the proteasome mechanism. Mechanically, we unraveled that CRYAB regulated FTH1 protein stability in a lactylation-dependent manner. Knockdown of FTH1 suppressed the osteogenic differentiation of BMSCs, and increased the cellular Fe and ROS levels, and eventually promoted ferroptosis. Rescue experiments revealed that CRYAB suppressed ferroptosis and promoted osteogenic differentiation of BMSCs via regulating FTH1. The mRNA level of FTH1 was decreased in the plasma of osteoporosis patients.

Conclusions: Downregulation of CRYAB boosted FTH1 degradation and increased cellular Fe and ROS levels, and finally improved the ferroptosis and lessened the osteogenic differentiation of BMSCs.