monocytic leukemia cell line thp1  (ATCC)

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet:

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: RNA sequencing, In Vitro, Ex Vivo

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet: A) Genetically predicted lower LACTB expression—instrumented using macrophage cis-eQTLs at the LACTB locus—is associated with lower AD risk (left panel). Genetically predicted lower LACTB expression is also associated with higher succinylcarnitine levels in the CSF (middle panel). Furthermore, genetically predicted higher succinylcarnitine levels are associated with lower AD risk (right panel). B) LACTB is expressed higher in the immune cluster compared to other brain cells, in contrast to another mitochondrial protein (SDHA), data from Brain Atlas . C, D) LACTB mRNA (C) and protein levels (D) are increased upon differentiation (THP1 macrophages vs monocytes and WTC11 iMGLs vs iPSC). E) Succinylcarnitine levels are increased in the lysate of LACTB KD/KO myeloid cells compared to SCR/WT. F) Succinylcarnitine is increased in the cell culture media of LACTB KD/KO myeloid cells compared to SCR/ WT. G) Higher succinylcarnitine levels in mice with lower LACTB expression (C57B6 x SLJ background, 2 months old). H) Higher succinylcarnitine levels in the liver, brain, isolated microglia and astrocytes of LACTB enzymatically-dead (ED) mice compared to WT (n=2 mice per genotype, 3 months old). I) LACTB protein levels increase with age in WT mice (12 vs 2 months old). J) Succinylcarnitine levels decrease with age in WT mice (12 vs 2 months old). Graphs display individual data points (left) alongside estimated marginal means with 95% confidence intervals (right). For the raw data, dot shapes represent independent macrophage differentiations, and dot colors indicate distinct microglia clones. Statistical details are provided in .

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: Expressing, Cell Culture, Isolation, Clone Assay

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet: A) Representative curves of succinyl-D-Asp substrate cleavage over time, showing strong reduction/abolishment in LACTB KD/KO myeloid cells compared to SCR/WT (a1: n=3 THP1 macrophages differentiations, a2: n=3 clones per genotype WTC11 iMGLs, a3: mouse BMDMs n=6 mice per genotype). Area under the curve (AUC) was calculated and used as outcome variable in statistical analyses. B) 13 C 3 -carnitine generation after substrate ( 13 C 7 -succinylcarnitine) incubation with LACTB recombinant protein in the absence or presence of LACTB inhibitor (Ac-IEPD-CHO) in a cell-free assay. C) Reduced label incorporation into carnitine in LACTB KO iMGLs after incubation with 13 C 3 -succinylcarnitine (n=3 clones per genotype) compared to WT WTC11 iMGLs. D) Representative heatmap showing label incorporation from 13 C 7 -succinylcarnitine into the indicated metabolites in WT iMGLs. E-J) LACTB is succinylated after treatment with diethyl-succinate (5 μM for 4 hours) in HEK cells (n=2 transfections, 4-10 well replicates each) (E) and THP1 macrophages (n=2 differentiations, 4 well replicates each) (H). Succinylation of LACTB is associated with a reduced rate of succinyl-D-Asp cleavage in HEK cells (n=2 transfections, 3 well-replicates each) (F) and THP1 macrophages (n=2 differentiations, 3 well-replicates each). AUC was calculated and used as outcome variable in statistical analyses. (I) LACTB succinylation is associated with higher succinylcarnitine levels in HEK cells (n=3) (G) and THP1 macrophages (n=3) (J). Graphs display individual data points (left) alongside estimated marginal means with 95% confidence intervals (right). For the raw data, dot shapes represent independent macrophage differentiations, and dot colors indicate distinct microglia clones. Statistical details are provided in .

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: Clone Assay, Incubation, Recombinant, Cell-Free Assay, Transfection

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet: A) Pathways enriched in GSEA analysis of bulk RNAseq data from LACTB KD THP1 macrophages compared to SCR (n=5 independent macrophages differentiations), NES = Normalized Enrichment Score. B) Pathways enriched in GSEA analysis of bulk RNAseq data from LACTB KO iMGLs compared to WT (3 clones per genotype, 2 differentiations per clone). C) Heatmap of Pearson correlation coefficients between LACTB expression and genes encoding pro-inflammatory cytokines across microglial states in publicly available human single-cell microglia datasets [ – ]. D) Single-cell clusters and corresponding annotations of WT and LACTB KO iMGLs (3 clones per genotype, 1-2 differentiations per clone). E) LACTB expression across clusters in WT iMGLs. F) Cluster proportions in WT and LACTB KO iMGLs. Cluster proportions were estimated using crumblr and compared by genotype using a generalized linear model with batch as a covariate (see Methods for further details). G) Pathways enriched in GSEA analysis of bulk RNAseq data from mouse microglia isolated from the brains of LACTB ED mice compared to controls (3 months old, n=3). Complete lists of genes and pathways are provided in and .

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: RNA sequencing, Clone Assay, Expressing, Single Cell, Isolation

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet: LACTB KD/KO in myeloid cells increases oxidative phosphorylation and reduces protein synthesis, cholesteryl esters, and triacylglycerides . A) Increased in OXPHOS (measured by mitostress seahorse assays) in LACTB KD THP1 macrophages (n=10 differentiations, 3-6 technical replicates each), and WTC11 LACTB KO iMGLs (3 clones per genotype, 3 differentiations per clone, 3-6 technical replicates each) compared to controls. B) Reduction in nascent protein synthesis (measured with a methionine analog) in LACTB KD/KO myeloid cells compared to SCR/WT. Graphs display individual data points (left) alongside estimated marginal means with 95% confidence intervals (right). For the raw data, dot shapes represent independent macrophage differentiations, and dot colors indicate distinct microglia clones. Full statistical details are provided in . C) Lipidomics alterations in LACTB KD vs SCR THP1 macrophages (n=3) and WTC11 LACTB KO vs WT iMGLs (n=3 clones, 1-2 differentiations each), showing a consistent reduction in CE and TG across the two myeloid cell types. Statistical details are provided in .

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: Phospho-proteomics, Clone Assay

Journal: bioRxiv

Article Title: Reduced LACTB expression in myeloid cells is associated with elevated succinylcarnitine levels and reduced Alzheimer’s disease risk

doi: 10.64898/2026.03.24.711053

Figure Lengend Snippet: A) LACTB mRNA expression increases after stimulation with IFN-β, IFN-γ, or TNF-α in THP1 macrophages (6 hours treatment) or WTC11 iMGLs (24 hours treatment). B) Succinylcarnitine levels decrease after stimulation with IFN-β, IFN-γ, or TNF-α in THP1 macrophages (6 hours treatment) or WTC11 iMGLs (24 hours treatment). C) Efferocytosis-related assays (myelin phagocytosis and lysosomal acidification, mass and proteolytic capacity) in LACTB KD/KO myeloid cells compared to SCR/WT. Graphs display individual data points (left) alongside estimated marginal means with 95% confidence intervals (right). For the raw data, dot shapes represent independent differentiations, and dot colors indicate distinct microglia clones. Statistical details are provided in .

Article Snippet: We used the monocytic human immortalized cell line THP1 (ATCC, TIB-202 RRID:CVCL_0006), which we treated with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml) for 72 hours to differentiate into macrophages.

Techniques: Expressing, Clone Assay