Exosome-transmitted HSPA9 facilitates bortezomib resistance by targeting TRIP13/USP1 signaling in multiple myeloma

Resistance to the proteasome inhibitor bortezomib (BTZ) poses a formidable therapeutic challenge in multiple myeloma (MM). Our study aims to analyze the mechanism by which exosomes heat shock 70 kDa protein 9 (HSPA9) secreted by BTZ-resistant MM cells disseminate resistance to BTZ-sensitive MM cells. The serum exosomes were identified by nanoparticle tracking analysis and transmission electron microscopy. Liquid chromatography-mass spectrometry and public databases were performed to screen exosomes HSPA9. Cell counting kit-8, western blotting and colony formation assay were used to detected the role of HSPA9 protein in vitro. Co-immunoprecipitation, immunofluorescence and protein truncation test experiments were used to determine the regulatory network of the HSPA9-USP1-TRIP13 complex. Optical imaging in vivo and xenograft mouse models were performed to investigate that exosomes HSPA9 promoted MM proliferation and BTZ resistance. We demonstrated that HSPA9 was highly expressed in serum exosomes and BTZ-resistant MM patients. Knockdown of HSPA9 significantly suppressed tumorigenesis and reversed BTZ resistance in vitro. As a downstream molecular of HSPA9, thyroid hormone receptor-interacting protein 13 (TRIP13) was also highly expressed in BTZ-resistant MM patients. Mechanistically, the carboxyl-terminal peptide-binding domain of HSPA9, provides a platform for recruiting the deubiquitinating enzyme ubiquitin-specific peptidase 1 (USP1), which prevents TRIP13 protein degradation. The HSPA9-USP1-TRIP13 complex exhibits stability in the cytoplasm, and its inhibition remarkably enhances BTZ resistance in vito. Our findings propose a pioneering molecular regulatory network in which MM-cell-derived exosomes HSPA9 transmitted BTZ resistance through the USP1/TRIP13 signaling pathway. This research highlights exosomes HSPA9 as a promising target to overcome MM BTZ resistance. Bortezomib (BTZ) is a proteasome inhibitor used to treat multiple myeloma (MM), however, resistance to BTZ poses a formidable challenge in MM treatments and the underlying mechanisms remain poorly understood. In this study, we demonstrated that the heat shock 70 kDa protein 9 (HSPA9) was highly expressed in the serum exosomes of BTZ-resistant MM patients and cell lines. Elevated levels of HSPA9 promoted disease progression, poor prognosis, cell proliferation, BTZ resistance and stemness characteristics in MM. In addition, exosomes HSPA9 secreted by BTZ-resistant MM cells can disseminate BTZ resistance characteristics to BTZ-sensitive MM cells via endocytosis and activated drug resistance signaling pathways at the same time. Mechanistically, the carboxyl-terminal peptide-binding domain of HSPA9 provided a platform for recruiting a deubiquitinating enzyme named ubiquitin-specific peptidase 1 (USP1), which could prevent thyroid hormone receptor-interacting protein 13 (TRIP13) protein from degradation. The HSPA9-USP1-TRIP13 complex exhibits stability in the cytoplasm, and its inhibition rendered MM cells more sensitive to BTZ treatment. Consequently, our findings propose a pioneer molecular regulatory network in which MM-cell-derived exosomes HSPA9 can transmit BTZ resistance through the USP1/TRIP13 signaling pathway. This research highlights exosomes HSPA9 as a promising therapeutic target to overcome BTZ resistance in MM.