Osteoarthritis (OA), a leading cause of chronic disability worldwide, is increasingly recognized to be driven by the accumulation of senescent chondrocytes (sCDs) and their deleterious pro-inflammatory senescence-associated secretory phenotype (SASP). Existing therapies, including senolytics and senomorphics, lack cell-specific targeting and fail to neutralize the heterogeneous components of SASP. Here, we develop a dual-engineered macrophage membrane camouflaged, self-assembled nanoplatform (BS@MD) that combines senolytic and senomorphic functions synergistically. Within the OA microenvironment, BS@MD acts as a "nanosponge" to broadly neutralize SASP through overexpressed cytokine receptors derived from LPS-primed macrophage membranes. This process alleviates chondrocyte senescence and facilitates the phenotypic shift of pro-inflammatory M1 macrophages toward an anti-inflammatory M2 state. Additionally, surface conjugation with an anti-DPP4 antibody enables BS@MD to selectively target sCDs and disassemble in the acidic lysosomal environment, releasing bortezomib (BTZ) and sabutoclax (Sab). These agents act synergistically to inhibit the NF-κB and BCL-2 pathways, thereby inducing sCDs apoptosis and suppressing SASP production, effectively disrupting the senescence-inflammation feedback loop. In the anterior cruciate ligament transection (ACLT)-induced OA mouse model and naturally aged OA mouse model, BS@MD enhances joint retention, reduces cartilage degradation and inflammation, and promotes cartilage homeostasis. Overall, this work pioneers a dual-pronged senotherapeutic strategy for non-surgical OA management.

Tannic acid-modified graphene oxide nanoplatform incorporating bortezomib as a combination chemo- and NIR-mediated photothermal oral cancer therapy.

Nanoparticle-Based delivery of proteasome inhibitors for glioblastoma Therapy: Strategies to overcome Blood-Brain barrier and therapeutic resistance.

Bortezomib (BTZ) is a proteasome inhibitor approved for the treatment of multiple myeloma. It is under investigation for use in the treatment of other solid tumors, such as breast and prostate cancers. BTZ is known for its high potency, limited aqueous solubility, and systemic toxicity. These characteristics restrict its clinical application. This study aims to formulate BTZ in the form of bovine serum albumin (BSA) nanoparticles (NPs) to enhance its anticancer targeting. The Box-Behnken experimental design was employed to optimize the formulation of BTZloaded BSA NPs. BTZ BSA NPs were prepared through the desolvation method by using glutaraldehyde (Glut) as a crosslinker. The Box-Behnken design was employed as an experimental design tool to investigate the impact of formulation parameters, specifically the amount of BTZ, BSA, and antisolvent volume, on outcomes, including particle size (PS), entrapment efficiency (EE%), and polydispersity index (PDI). The morphology of the prepared NPs was examined using a transmission electron microscope (TEM), while a compatibility study was conducted using Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The optimized formulation resulted in a mean PS, EE%, and PDI of 74 nm, 68%, and 0.007, respectively. The BTZ BSA NPs exhibited a sustained in vitro release profile over 24 hours. TEM revealed that the prepared NPs had an almost spherical morphology. FTIR and DSC revealed that BTZ was encapsulated well within the prepared NPs with minimal decomposition and/or degradation. The optimized BTZ BSA NPs exhibited uniform nanosize, high entrapment efficiency, and sustained drug release, confirming the effectiveness of Box-Behnken optimization. Encapsulation within the cross-linked albumin matrix protected BTZ from hydrolytic degradation, enhanced its stability, and provided controlled release, supporting its potential for improved anticancer activity and reduced systemic toxicity. The use of experimental design is a valuable tool for optimizing the formulation of BTZ BSA NPs, achieving the required PS, high EE%, and low PDI.

Background: Stromal remodeling in the tumor microenvironment contributes to multiple myeloma (MM) progression and drug resistance, but the extracellular mediators that drive these changes remain incompletely defined. Extracellular enolase-1 (ENO1), including membrane-associated and secreted forms, has been implicated in tumor progression; however, its role in cancer-associated fibroblast (CAF)-associated stromal reprogramming in MM is unclear. Methods: The effects of extracellular ENO1 on stromal activation and tumor-supportive functions were examined in MM using MM-bone marrow stromal cell (BMSC) co-cultures, lactate production and viability assays, immunoblotting, cytokine analyses, and a subcutaneous xenograft model of bortezomib (BTZ)-resistant MM in male 6-7-week-old NOD.Cg-Prkdcscid Il2rgtm1Vst/Vst (NPG) mice. HuL001, an anti-ENO1 monoclonal antibody, was used to evaluate the therapeutic relevance of extracellular ENO1 targeting. Results: Extracellular ENO1 promoted fibroblast activation protein expression through plasmin-mediated transforming growth factor-β (TGF-β) activation and induced a CAF-associated stromal phenotype marked by enhanced glycolytic activity and increased secretion of tumor-promoting cytokines in MM-BMSC co-cultures. HuL001 suppressed these ENO1-driven effects. HuL001-pretreated stromal cells also exhibited reduced tumor-supportive activity in a BTZ-resistant MM xenograft model. In addition, HuL001 combined with lenalidomide overcame BTZ resistance in MM. Conclusions: Extracellular ENO1 drives CAF-associated stromal reprogramming in the MM microenvironment through the ENO1/plasminogen/plasmin/TGF-β axis. Therapeutic targeting of extracellular ENO1 with HuL001 may disrupt these tumor-supportive stromal activities and help overcome drug resistance in MM.

RAD23A promotes multiple myeloma cell survival through DNA damage response, proteostasis and enhanced metabolic activity.

Inter-individual variability in patient susceptibility to bortezomib (BTZ)-induced peripheral neuropathy (BIPN) suggests a potential role of genetic predisposition. However, the comprehensive mutational landscape and its functional relevance remain poorly defined. We aimed to characterize the genetic architecture and inflammatory mechanisms underlying BIPN in multiple myeloma (MM) patients. Whole exome sequencing (WES) was performed on peripheral blood mononuclear cells (PBMCs) from 20 newly diagnosed MM patients treated with BTZ, including eight patients who developed grade ≥ 2 BIPN and 12 controls without neuropathy. To functionally interpret genetic alterations, WES data were integrated with transcriptomic datasets from chemotherapy-induced peripheral neuropathy models obtained from GEO databases. Enrichment analyses and molecular docking were conducted to identify key driver genes and potential BTZ-protein interactions. WES identified 90,465 BIPN-associated single nucleotide polymorphisms, with a distinct co-mutation signature involving 33 zinc-finger (ZNF) family genes. Integrative multi-omics analysis yielded 100 candidate genes enriched in inflammatory response, neuronal development, synaptic organization, and MAPK/NF-κB signaling pathways. Three key genes-CACNA1H, CIC, and ABLIM2-were identified as potential driver genes and demonstrated direct binding affinity with BTZ in docking analyses. Notably, enrichment analyses also suggested shared molecular mechanisms between neurotoxicity and cardiotoxicity. Our findings revealed that inflammation-driven neuronal dysfunction mediated by genetic susceptibility represented a central mechanism of BIPN. Integrative genomic profiling might provide a framework for personalized risk assessment and precision management of BTZ-related neurotoxicity and associated organ toxicity.

Bortezomib (BTZ), the first-generation proteasome inhibitor, has been approved for the treatment of relapsed, refractory, and newly diagnosed multiple myeloma. Despite its remarkable antitumor efficacy, BTZ treatment is severely limited by a high incidence of systemic adverse reactions, primarily due to its non-selective cytotoxicity toward rapidly dividing normal cells and its potent neurotoxic effects on peripheral neurons. Bortezomib-induced peripheral neurotoxicity (BIPN) manifests as neuropathic pain and sensory abnormalities, affecting up to 31% to 64% of patients and limiting BTZ's clinical use. Currently, the underlying mechanisms of BIPN are poorly understood. To evaluate the effects of BTZ on the functions of peripheral nerves in mice, we administered an intraperitoneal injection treatment for four weeks. Results indicated that BIPN caused mechanical allodynia, gait abnormalities, and pathological changes in myelin and axons in mice. This study confirms that BTZ upregulates the expression of the activating transcription factor 3 (ATF3), which in turn mediates the increased expression of the copper transporter SLC31A1, causing dysregulation of intracellular copper ion homeostasis and subsequent copper accumulation, and ultimately inducing the development of peripheral neurotoxicity. Elevated intracellular copper concentration exerts a dual effect: it directly promotes the oligomerization of Dihydrolipoamide S-acetyltransferase (DLAT) and concurrently damages the iron-sulfur cluster protein ferredoxin 1 (FDX1), collectively triggering the onset of cuproptosis. Green tea has garnered attention for its rich content of catechins, with (-)-Epigallocatechin Gallate (EGCG) being the most abundant catechin present. This study uncovers the molecular mechanism by which EGCG inhibits BTZ-induced cuproptosis through targeted regulation of copper homeostasis. Analyses demonstrate that EGCG significantly downregulates the expression of the copper transporter SLC31A1, thereby effectively suppressing transmembrane influx of extracellular copper ions. This intervention markedly reduces intracellular copper overload, eliciting a dual regulatory effect: on one hand, the decreased copper concentration directly inhibits the oligomerization of DLAT; on the other hand, it effectively protects the iron-sulfur cluster protein FDX1 from damage. This study aims to systematically elucidate the molecular mechanisms underlying BIPN and to evaluate the therapeutic potential of EGCG in alleviating BIPN, offering a novel therapeutic strategy for the prevention and treatment of BIPN.

Abstract Background and Purpose Sepsis induces immunosuppression, contributing to high mortality. Plasmablasts, including an IL‐10–producing regulatory subset, expand during sepsis to exert immunosuppressive functions. This study examined how depleting plasmablasts can improve the prognosis of sepsis, and investigated the underlying mechanisms by focusing on neutrophil function. Experimental Approach Plasmablasts were depleted in vivo in CD138‐diphtheria toxin receptor (DTR) transgenic mice subjected to caecal ligation and puncture (CLP)‐induced sepsis. The effects of plasmablast depletion on animal survival, bacterial load, inflammatory mediators, and neutrophil number and function were assessed. The proteasome inhibitor bortezomib (BTZ) was evaluated for its ability to suppress plasmablasts, and its therapeutic efficacy in wild‐type mice with sepsis. Key Results Plasmablast depletion in CD138‐DTR mice was associated with an increase in the number and antibacterial function of neutrophils, including increased production of reactive oxygen species (ROS) and phagocytic capacity. In vitro co‐culture experiments demonstrated that plasmablasts suppressed neutrophil activity, an effect mediated by adenosine and IL‐10 signalling. Treatment of wildtype septic mice with BTZ effectively reduced plasmablast abundance and CD39 expression. BTZ 0.05 mg·kg −1 mirrored the protective effects of genetic ablation, improved survival, reduced bacterial burden and inflammation and improved neutrophil‐mediated bacterial clearance. At high concentrations, BTZ worsened outcomes and is an unlikely magic bullet for sepsis without stringent dosing controls. Conclusion and Implications Plasmablasts contribute to sepsis immunosuppression by affecting neutrophil function. The clinically‐available drug bortezomib can target this population, positioning plasmablast inhibition as a novel and translatable therapeutic strategy to improve bacterial clearance and survival in sepsis.

The metabolic-epigenetic symbiosis between tumor cells and macrophages in the bone marrow microenvironment (BMM) plays a crucial role in immune evasion and therapeutic resistance in multiple myeloma. Here, we present a copper-based nanomodulator, NanoCURE (Cu-activated Reprogramming Eraser), that targets the glycolysis-lactate-lactylation axis to reprogram metabolism and epigenetics in the BMM. To construct NanoCURE, lactate oxidase (LOx) and bortezomib (BTZ) are coencapsulated within a tumor-activated Cu2+ nanoassembly, facilitating bone marrow (BM)-specific delivery via an in vivo hijacking monocyte/macrophage pathway. Mechanistically, NanoCURE acts as a multifunctional modulator that disrupts the metabolic-epigenetic positive feedback loop by directly blocking histone lactylation through site-specific binding while simultaneously suppressing the upstream Akt/mTOR/c-Myc signaling axis. Moreover, NanoCURE can trigger the overproduction of reactive oxygen species (ROS), leading to mitochondrial dysfunction that amplify epigenetic interference. Consequently, these synergistic effects effectively disrupt the metabolic and epigenetic support of MM and reverse immunosuppressive M2 macrophage polarization to enhance the therapeutic effect of proteasome inhibitors in an orthotopic xenograft mouse model. Furthermore, NanoCURE achieves precise bone marrow enrichment via monocyte hijacking while maintaining low systemic copper levels, thereby ensuring high biosafety, preserving hematopoietic integrity, and exhibiting no observable organ toxicity. In summary, this work introduces a carrier-as-drug platform that targets the glycolysis-lactate-lactylation axis to enable in situ metabolic-epigenetic-immune reprogramming, offering a promising strategy to overcome therapeutic resistance in multiple myeloma.

Multiple myeloma (MM) is a hematologic malignancy commonly treated with bortezomib (BTZ). However, treatment efficacy is often limited by the development of BTZ resistance. Icaritin has demonstrated broad anti-tumor activities. This study aimed to investigate the effect of Icaritin on the malignant progression of MM and its potential to overcome BTZ resistance. The anti-MM activity of Icaritin was evaluated through a series of experimental approaches, including cell counting kit 8, flow cytometry, 5-ethynyl-2'-deoxyuridine, network pharmacology, molecule docking, reverse transcription-quantitative polymerase chain reaction, Co-Immunoprecipitation (CoIP), and western blot. Icaritin suppressed MM cell viability and proliferation, induced ferroptosis, and enhanced cellular sensitivity to BTZ. Moreover, Icaritin inhibited HSP90α family 1 (HSP90AA1) protein expression via promoting ubiquitination. HSP90AA1 facilitated MM cell proliferation, suppressed ferroptosis, and attenuated BTZ sensitivity. Notably, Icaritin promoted MM cell ferroptosis and BTZ sensitivity by inhibiting HSP90AA1. Invivo, Icaritin enhanced the sensitivity of tumor cells to BTZ. Icaritin suppresses malignant progression, induces ferroptosis, and enhances BTZ sensitivity in MM by inhibiting HSP90AA1. These findings provide a novel theoretical basis for Icaritin treatment of MM and regulation of BTZ sensitivity.

Bortezomib (BTZ) is clinically important in the nephrological field because of its increasing use in plasma cell disorders and antibody-mediated kidney diseases, where it can both exert therapeutic benefits and, paradoxically, cause significant renal toxicity. This study investigated the protective effects of vanillic acid (VA) against BTZ-induced acute kidney injury using biochemical and molecular approaches. BTZ administration elevated serum creatinine, blood urea nitrogen, KIM-1 and NGAL, while co-treatment with VA partially normalized these markers. BTZ increased apoptotic markers (BAX, PUMA and TRAIL) and inflammatory cytokines (IL-1β, IL-6, TNF-α and IL-10), which were attenuated by VA. Oxidative stress-related genes NQO1, NOX4 and XO were upregulated, and GPX4 was downregulated by BTZ; VA restored these expressions. BTZ disrupted mitochondrial dynamics and energy metabolism (MFN2, CPT1A and OxPhos decreased; FIS1 increased), with VA ameliorating these changes. Energy imbalance induced by BTZ, reflected by reduced ATP and increased LDH and TAG, was also mitigated by VA. Podocyte proteins nephrin, podocin and CD2AP were reduced, accompanied by increased LAMP1 and decreased miR-204-5p; VA partially restored these levels. Overall, VA protected against BTZ-induced kidney injury via antioxidant, anti-inflammatory, antiapoptotic and mitochondrial mechanisms, potentially involving the miR-204-5p-nephrin axis.

A ketogenic diet (KD) can benefit a range of neurological conditions, including different types of peripheral neuropathy. Bortezomib (BTZ), a first-line chemotherapy for treating multiple myeloma, has neurotoxic effects and often induces chemotherapy-induced peripheral neuropathy. Here, we tested whether a KD could prevent the onset of small fiber neuropathy and behavioral hypersensitivity associated with bortezomib-induced peripheral neuropathy (BIPN). Male C57BL/6 mice were fed either a standard chow diet or a KD for 10 days. During the first 5 days of KD administration, mice received daily intraperitoneal injections of BTZ or vehicle. Mice entered a BTZ washout period on day 6 while remaining on their specific diets. Hindpaw responses to mechanical (von Frey) and cold (acetone) stimuli were measured to determine how KD and BTZ treatment affected behavioral sensitivity. Footpads were harvested for analysis of intraepidermal nerve fiber density (IENFD), and lumbar dorsal root ganglia were collected for in vitro analysis of neurite outgrowth and cellular bioenergetics. Bortezomib-treated, KD-fed mice exhibited decreased mechanical hypersensitivity, normalized cold hypersensitivity, preserved IENFD, and decreased extracellular acidification rate compared with BTZ-treated, chow-fed mice. In vitro analysis of ketones' ability to protect against BTZ-induced reductions in neurite growth revealed that cultured neurons pretreated with ketones were protected from BTZ-induced neurite degeneration. Findings suggest that KD protects against reductions in IENFD, behavioral hypersensitivity, and bioenergetic shifts induced by BIPN, and provides evidence that KD provides protection against BTZ and could be a potential therapeutic for BIPN.

Multiple chemotherapeutic agents exhibit neurotoxicity. For example, bortezomib (BTZ)-induced peripheral neuropathy (BIPN) is characterized by sensory abnormalities and pain, and effective treatment strategies are currently lacking. This study aimed to investigate the alleviating effects of gastrodin (GAS) on BIPN and its potential mechanisms. Behavioral tests were used to assess changes in pain thresholds across all groups of mice. Hematoxylin-eosin staining, transmission electron microscopy, and immunofluorescence were used to evaluate peripheral nerve injury and the activation of spinal glial cells. ELISA was used to measure the levels of inflammatory cytokines. Proteins associated with the NF-κB/NLRP3 pathway were examined using Western blot. GAS significantly improved thermal and mechanical hyperalgesia in BIPN mice. Moreover, BTZ can cause loss of intraepidermal nerve fibers, microstructural damage to the dorsal root ganglia, a disorganized arrangement of sciatic nerve fibers, and demyelination, all of which were effectively reversed by GAS treatment. Further investigation revealed that GAS significantly suppressed the upregulation of IBA-1 and GFAP in the spinal cord of BIPN mice, and the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were concurrently reduced. IBA-1/IL-1β and IBA-1/TNF-α double-labeled positive cells were significantly increased in BIPN mice, and GAS intervention reduced the number of these double-labeled cells. In addition, GAS significantly inhibited aberrant NF-κB signaling and upregulation of NLRP3 inflammasome-related proteins of BIPN mice. GAS may alleviate BIPN by suppressing microglia-mediated neuroinflammatory responses, and the NF-κB/NLRP3 inflammasome signaling pathway appears to be involved in this process.

Purpose: Bortezomib (BTZ) is widely used in multiple myeloma (MM), but its toxicity shows marked interindividual variability. This study aimed to identify pharmacogenetic and clinical factors associated with BTZ-related adverse drug reactions (ADRs). Methods: A retrospective and prospective observational study was conducted in 127 MM patients treated with BTZ-based regimens. Polymorphisms in CYP enzymes and ABCB1 were genotyped using qPCR. Associations between genetic variants, treatment response, and ADRs were assessed using univariate and multivariate analyses with Benjamini-Hochberg correction. Results: ADRs occurred in 98.4% of patients, most commonly gastrointestinal toxicity (49%), general toxicity (46%), and peripheral neuropathy (39%). Women showed higher rates of gastrointestinal toxicity and non-peripheral neurotoxicity. Multivariate analysis identified the ABCB1 C1236T A/G genotype as protective against gastrointestinal toxicity, while the CYP3A4 intermediate metabolizer phenotype was associated with increased psychiatric toxicity. TP53 mutations were independently associated with hematologic and renal toxicity. Kaplan-Meier analysis showed earlier onset of peripheral neuropathy and respiratory toxicity in CYP3A4 intermediate and poor metabolizers. Conclusions: Genetic variation in ABCB1 and CYP3A4, together with clinical factors such as TP53 mutation and sex, may contribute to interindividual variability in BTZ safety in MM. These findings should be considered exploratory given the sample size and require confirmation in larger cohorts. Nonetheless, they suggest a potential role for pharmacogenomics in supporting future approaches to treatment personalization.

Conventionally, KDM5C functions as a specific demethylase that targets histone H3 lysine 4 dimethyl and trimethyl modifications, crucial for gene expression. However, the role of KDM5C in multiple myeloma (MM) progression and bortezomib (BTZ) resistance has remained elusive. In this study, we found noncanonical functions of KDM5C in MM. Specifically, KDM5C binds to CBP and MYC, conferring BTZ resistance in MM through a demethylase-independent mechanism. Our investigations revealed that KDM5C is markedly upregulated in BTZ-resistant MM patients as well as those with relapsed MM. Significantly, the expression level of KDM5C exhibits an inverse correlation with the overall survival of MM patients. Moreover, KDM5C is indispensable for MM cell proliferation. Depletion of KDM5C augmented the sensitivity of MM cells to BTZ treatment both in vitro and in vivo. We found that KDM5C forms a novel complex with CBP and MYC via its PHD2 domain. This complex formation triggers lysine 27 acetylation in histone H3 (H3K27ac) and subsequent enrichment of H3K27ac on the PERK promoter. As a result, PERK transcription is activated, and Nrf2 phosphorylation is promoted, bolstering the unfolded protein response within the endoplasmic reticulum of MM cells. In contrast, the methylation status of histone H3 lysine 4 (H3K4me1/3) on the PERK promoter remains unaltered, regardless of the complex state. Taken together, the findings of this study underscore the key role of KDM5C as a driving force behind MM progression and BTZ resistance, indicating that KDM5C represents a novel and promising therapeutic target for the treatment of BTZ-resistant MM.

Introduction: Bortezomib (BTZ), a proteasome inhibitor, significantly improved multiple myeloma (MM) treatment; however, drug resistance remains a major challenge, limiting its effectiveness. Understanding the molecular basis of this resistance is crucial for developing new therapies and overcome the resistance issue. Long-read RNA sequencing offers a powerful method to thoroughly characterize the transcriptome, including full-length transcripts and splice variants, potentially revealing previously unknown mechanisms of bortezomib resistance.Methods: Total RNA was extracted from BTZ-resistant and BTZ-sensitive clones from two MM cell lines (AMO and H929), in three replicates. After appropriate library preparation, long-reads RNAseq was performed using Oxford Nanopore technology, on the PromethION platform. All the quality control analyses were carried out and transcript were annotated, producing an expression matrix. The R package IsoformSwitchAnalyzeR was used to analyze isoform expression and differential isoform usage of the different transcripts among the conditions.Results: The Bioinformatic analyses carried out on AMO cell line made possible to observe 53 significant switches (FDR<0.05; absolute dIF>0.1) affecting 45 different genes, involving a total of 93 isoforms. Moreover, two of these genes coded for proteasome subunits (PSMA6, PSMB10).The H929 cell line was affected by 41 significant switches on 37 genes, with 76 isoforms involved. In both cases, the BTZ-resistant clones showed a significant upregulation of the transcript ENST00000570985, coding for a Nonsense Mediated Decay (NMD)-insensitive variant of the PSMB10 gene, suggesting the potential role of non-coding transcripts in the molecular mechanisms of drug resistance. 18 switching genes were shared between both the BTZ-resistant clones of MM cell lines.The switching genes were further analysed investigating their ontology and their involvement in pathways through pathway enrichment analysis conducted on Reactome. These analyses highlighted several genes coding for histones (H2AC6, H4C15, H2BC4, H2AZ2, H1-2,H2BC4). Two transcriptomic variants referred to the H2AC6 gene were observed, with a significant increase in the NMD-sensitive transcript ENST00000314088 for the resistant cells, in which almost all the expression is dependent on this non-coding variant, suggesting the relative absence of the related protein. Of note, H2AC6 represents an elite gene related hematologic cancer according to Cancer Gene Census (Disease Ontology: DOID:2531)Conclusions: Based on what observed, long-reads RNAseq represents an important strategy to better understand isoform usage of specific genes, attributing their overall expression to specific transcript variants, paving the way for a better comprehension of molecular mechanisms underlying resistance to therapy. The high proportion of shared switching genes between BTZ-resistant cell lines, suggests a potential mechanism based on this process.

Introduction. In multiple myeloma (MM) treated with proteasome inhibitors, cells can rewire carbon flow to survive proteotoxic stress. We generated bortezomib (BTZ)–resistant (RES) clones by chronic stepwise exposure and compared them with sensitive (SENS) counterparts in AMO and NCI-H929 cell lines. Methods. Proteomics was performed by LC–MS/MS on an Orbitrap platform, targeted metabolomic was performed by LC-MS. RNA sequencing used the Nanopore workflow and key findings were validated by Western blot and RT-qPCR. Results. RNA sequencing of both cell lines revealed a consistent metabolic rewiring in resistant cells. These cells exhibited broadly increased expression of respiratory-chain subunits and tricarboxylic acid cycle enzymes, indicating a heightened oxidative capacity. Concurrently, pyruvate carboxylase, the key enzyme for pyruvate-to-oxaloacetate anaplerosis, was significantly reduced. This metabolic shift was accompanied by elevated expression in glycolytic/pentose phosphate pathway and serine synthesis pathway modules. Furthermore, amino acid handling nodes were upregulated alongside MAT2A, suggesting enhanced one-carbon and S-adenosylmethionine support, consistent with an adaptive ATF4-driven integrated stress response. Moreover, heatmaps showed coordinated increases in amino-acid/ISR nodes including ASNS, SLC7A11 (xCT), SLC3A2/LAT, GLS, GLUD1, GOT1, HSPA5/XBP1, SQSTM1, and antioxidant enzymes (GCLC/NQO1/TXNRD2/HMOX1), with MAT2A suggesting one-carbon/SAM support. AMO cells showed decreased intracellular glutamine with a trend to elevated glutamate, while H929 cells displayed a significant rise in glutamate without a change in glutamine—compatible with fast Gln→Glu conversion buffered by continued uptake. Functionally, glutamine deprivation caused a sharper viability drop and lactate modulation in RES, confirming dependency. Importantly, comparing RES + glutamine vs RES in Gln-free revealed activation of SREBF/SREBP-driven programs and the cholesterol-biosynthesis superpathway, alongside reinforcement of TCA/FA β-oxidation and NRF2-mediated oxidative-stress response, indicating that lipid/sterol and redox metabolism are integral to the glutamine-supported resistant state. From UCSC Xena (MMRF-COMMPASS) we retrieved a Kaplan–Meier curve stratified by GLUD1 and GLUD2 expression. Patients with GLUD1-high tumors exhibit significantly inferior overall survival, reinforcing the importance of the glutaminolysis-driven anaplerotic program in MM progression. Conclusions. Together, these data define an anaplerotic rerouting model, PC down-tuning limits pyruvate-derived OAA and shifts TCA fueling toward glutaminolysis. This maintains high respiratory capacity while preserving a favorable NADPH/GSH buffer and an adaptive UPR/autophagy support and one-carbon–lipid coupling that together stabilize the resistant phenotype. Targeting this metabolic vulnerability offers a promising strategy to overcome bortezomib resistance in multiple myeloma.

Proteasome inhibition by bortezomib augments the efficacy of anti-PD-L1 therapy against lung cancer.

Triple-negative breast cancer (TNBC) demonstrates elevated death rates in its advanced stage. Bortezomib (BTZ) and Noscapine (NCP) have potential as antitumor agents in various cancers, including TNBC. In this investigation, we present a way to enhance the therapeutic outcomes in TNBC by utilizing BTZ and NCP, employing poly(D,L-Lactide-coglycolide, PLGA). BTZ and NCP, which exhibit lower water solubility, were co-encapsulated in PLGA nanoparticles (BN-PLGA) and stabilized with 0.5% PVA, resulting in a low PDI and greater homogeneity. Several spectroscopic techniques were used to analyze BTZ- and NCP-loaded PLGA (BN-PLGA NPs). The in vitro release of these drugs from BN-PLGA NPs was examined at physiological and acidic pH to evaluate the drug release process. The effectiveness of BN-PLGA NPs against TNBC (MDA-MB-231) was assessed. In vitro drug release at pH 7.4 and 5.0 showed that BTZ and NCP encapsulated in PLGA NPs containing 0.5% PVA were sustained, unlike the free drugs, which showed a relatively rapid release. BN-PLGA NPs containing 0.5% PVA were spherical, with a mean diameter of 223nm, and exhibited high encapsulation efficiency (BTZ ∼ 83%, NCP ∼ 78%) and enhanced cytotoxicity relative to BTZ monotherapy in vitro. The BN-PLGA NPs showed better MICs against P. aeruginosa, S. epidermidis, and S. aureus than the pure drug. The findings indicate that combining NCP and BTZ via PLGA NPs could be a viable approach for TNBC treatment, potentially paving the way for extensive in vivo investigations to assess the safety and efficacy of these innovative nanoformulations using TNBC animal models.