Effect and mechanism of BYL-719 on Mycobacterium tuberculosis-induced differentiation of abnormal osteoclasts

doi:10.12307/2024.833

Abstract

Abstract: BACKGROUND: The phosphatidylinositol 3-kinase/protein kinase (PI3K/AKT) signaling pathway plays a pivotal role in regulating osteoclast activation, which is essential for maintaining bone homeostasis. Bone destruction in osteoarticular tuberculosis is caused by aberrant osteoclastogenesis induced by Mycobacterium tuberculosis infection. However, the role of the PI3K signaling pathway in Mycobacterium tuberculosis-induced aberrant osteoclastogenesis remains unclear.
OBJECTIVE: To investigate the effects and mechanisms of the PI3K/AKT signaling pathway inhibitor BYL-719 on aberrant osteoclastogenesis induced by Mycobacterium tuberculosis.
METHODS: RAW264.7 cells were infected with bovine Mycobacterium tuberculosis bacillus calmette-cuerin vaccine, and Ag85B was used for cellular immunofluorescence staining. The cell counting kit-8 assay was employed to determine the safe concentration of BYL-719. There were four groups in the experiment: blank control group, BYL-719 group, BCG group, and BCG+BYL-719 group. Under the induction of receptor activator of nuclear factor kappa-B ligand, the effects of BYL-719 on post-infection osteoclast differentiation and fusion were explored through tartrate-resistant acid phosphatase staining and phalloidin staining. RT-PCR and western blot were used to detect the expression of osteoclast-related genes and proteins, and further investigate the mechanism of action.
RESULTS AND CONCLUSION: Immunofluorescence staining showed that RAW264.7 cells phagocytosed Mycobacterium tuberculosis. Cell counting kit-8 data indicated that 40 nmol/L BYL-719 was non-toxic to cells. Tartrate-resistant acid phosphatase staining and phalloidin staining showed that BYL-719 inhibited the generation and fusion ability of osteoclasts following infection. RT-PCR and western blot results also indicated that BYL-719 suppressed the upregulation of osteoclast-specific genes (including c-Fos, NFATc1, matrix metalloproteinase 9, and CtsK) induced by Mycobacterium tuberculosis infection (P < 0.05). Western blot and immunofluorescence staining revealed that BYL-719 inhibited excessive osteoclast differentiation induced by Mycobacterium tuberculosis by downregulating the expression of IκBα-p65. To conclude, BYL-719 inhibits aberrant osteoclastogenesis induced by Mycobacterium tuberculosis through the downregulation of IκBα/p65. Therefore, the IκBα/p65 signaling pathway is a potential therapeutic target for osteoarticular tuberculosis, and BYL-719 holds potential value for the preventing and amelioration of bone destruction in osteoarticular tuberculosis. BYL-719 has the potential to prevent and ameliorate bone destruction in osteoarticular tuberculosis.

Key words: BYL-719, Mycobacterium tuberculosis infection, osteoclast, osteoarticular tuberculosis, PI3K/AKT, RAW264.7, bacillus calmette-cuerin vaccine, NF-κB

CLC Number:

Zhang Jun, Guo Jian, Jia Qiyu, Tang Lili, Wang Xi, Abudusalamu · Alimujiang, Wu Tong, Maihemuti · Yakufu, Ma Chuang. Effect and mechanism of BYL-719 on Mycobacterium tuberculosis-induced differentiation of abnormal osteoclasts [J]. Chinese Journal of Tissue Engineering Research, 2025, 29(2): 355-360.

Figures/Tables 8

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