The ACOD1 Knockout 143B Polyclonal Cells provide a genetically engineered human osteosarcoma cell population for studying ACOD1/IRG1 function in immunometabolism and cancer biology. Generated via CRISPR/Cas9-mediated gene disruption, this polyclonal pool introduces heterogeneous loss-of-function mutations across the ACOD1 locus, enabling robust modeling of ACOD1 deficiency without clonal selection artifacts. The product offers a versatile platform for investigating itaconic acid biosynthesis, macrophage-mediated immune regulation, and tumor microenvironment interactions in a bone tumor context.
The parental 143B cell line is a highly tumorigenic and metastatic osteosarcoma derivative of the HOS lineage, selected for its aggressive in vivo behavior. These cells model human osteosarcoma with high fidelity, recapitulating key features of bone tumor growth, invasion, and metastatic dissemination. Their mesenchymal origin and transformed phenotype make them a relevant system for examining tumor cell-intrinsic and microenvironmental contributions to cancer progression.
ACOD1 encodes aconitate decarboxylase 1, the enzyme that converts the TCA cycle intermediate cis-aconitate into the immunomodulatory metabolite itaconic acid. This reaction is strongly induced by inflammatory signals through NF-??B and IRF1 downstream of TLR4, IFN-??, and TNF-??. Itaconic acid then inhibits succinate dehydrogenase (SDH) of mitochondrial complex II, altering succinate levels and restricting cellular respiration. Concurrently, it modulates NLRP3 inflammasome activation and promotes an antioxidant response via KEAP1-dependent activation of NRF2. Through these axes, ACOD1 shapes macrophage effector phenotypes, dampens pro-inflammatory cytokine production, and exerts direct antimicrobial activity against intracellular pathogens.
In the context of 143B osteosarcoma cells, ACOD1 knockout is particularly relevant for dissecting the role of itaconic acid in tumor-associated immunosuppression. Osteosarcoma microenvironments often harbor macrophage populations with altered metabolic profiles; ACOD1-driven itaconic acid production may contribute to immune evasion by suppressing T cell function and fostering a tolerogenic niche. The 143B model permits investigation of how tumor cell-derived or recruited macrophage-derived itaconic acid influences tumor growth, metastasis, and response to immunotherapies in bone cancer settings.
Researchers can employ these ACOD1 knockout 143B polyclonal cells in a diverse array of experiments, including metabolic flux analyses to trace itaconic acid production by LC-MS, SDH activity assays, succinate quantification, and RT-qPCR profiling of inflammatory cytokine expression. Functional readouts such as NLRP3 inflammasome activation, NRF2 target gene induction, and macrophage marker expression can be assessed via western blotting and flow cytometry. Additionally, the cells are suitable for migration, invasion, and co-culture assays to explore tumor-immune interactions. For further details, please contact Ascent Research.
