Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: CTPS1 promotes the progression of DLBCL. (A) Immunoblot analysis showing CTPS1 expression in tumor cells isolated from DLBCL patients compared with purified normal CD19 + B cells derived from healthy donors (n = 3). (B) Immunoblot analysis of SUDHL4 and SUDHL8 showing the knockout efficiency of sgCTPS1 and expression of CTPS2, PARP, cleaved PARP and BCL-XL. (C) Quantitative reverse transcription PCR analysis of SUDHL4 and SUDHL8 showing the knockout efficiency of sgCTPS1 in DLBCL cells. Levels in cells with sgControl were set to 1. (D) Growth curve analysis of SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. (E) Flow cytometry analysis showing cell apoptosis in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs using annexin V-APC/7-AAD Kit. (F) The relative levels of nucleotide metabolites in SUDHL4 expressing control or CTPS1 sgRNAs detected by LC-MS. (G) Growth curve analysis of SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs, treated without or with CTP every 2 days. (H) Flow cytometry analysis showing cell apoptosis in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs using annexin V-APC/7-AAD Kit, treated without or with CTP. (I) Schematic diagram of SUDHL4 or SUDHL8 xenograft mouse model. (J, K) Western blot showing the efficiency of CTPS1 knockout, tumor growth curves and volume analysis of tumor in SUDHL4 and SUDHL8 xenograft mouse (n = 5 per group). Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Isolation, Purification, Derivative Assay, Knock-Out, Reverse Transcription, Control, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy

Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: CTPS1 promotes mitochondrial function and enhances mitophagy in DLBCL. (A) KEGG pathways showing the enrichment in SUDHL4 expressing control or CTPS1 sgRNAs by RNA sequencing (RNA-seq) (n = 3 per group). (B) Normalized expression of mitochondria encoded genes in SUDHL4 expressing control or CTPS1 sgRNAs by RNA-Seq. (C) Immunoblot analysis showing the expression of LC3 I/II in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. n = 3. (D) Immunoblot analysis showing the expression of TOM20 and TIM23 in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. n = 3. (E) Flow cytometry with MitoTracker in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. (F) Oxygen consumption rate (OCR) measurements in SUDHL4 expressing control and CTPS1 sgRNA. Arrows denote sequential addition of 1.5 μM oligomycin, 2 μM FCCP, and 0.5 μM rotenone/antimycin. (G) Mitochondrial superoxide (MitoSOX) analysis in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. (H) Mitochondrial membrane potential analysis in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs without or with 10 μM CCCP by JC-1. (I) MitoPerOx analysis measuring lipid peroxidation in SUDHL4 expressing control or CTPS1 sgRNAs (J) Immunoblot analysis showing the expression of DRP1, p -DRP1S616 and OPA1 in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. n = 3. (K) Representative images of mitochondria in the subcutaneous tumor of xenograft mice model was constructed by using SUDHL4 and SUDHL8 cells following control or CTPS1 sgRNAs expression by transmission electron microscopy (TEM). (L) Immunoblot analysis of LC3 I/II expression in the mitochondria in SUDHL4 expressing control or CTPS1 sgRNAs without or with 20 nM Bafilomycin A1 (BafA1). n = 3. (M) Immunoblot analysis showing the expression of BNIP3, PINK1 and Parkin in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. n = 3. (N) Immunofluorescence analysis showing BNIP3 expression around mitochondria in SUDHL4 expressing control or CTPS1 sgRNAs without or with 10 μM CCCP. Scale bar: 20 μm. Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: Expressing, Control, RNA Sequencing, Western Blot, Flow Cytometry, Membrane, Construct, Transmission Assay, Electron Microscopy, Immunofluorescence

Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: CTPS1 induces phospholipid metabolic remodeling in DLBCL. (A) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealing the upregulated enrichment pathways of DLBCL in the DepMap database (B) KEGG pathway enrichment of differential metabolites in SUDHL4 expressing control or CTPS1 sgRNAs by metabolomics analysis. (C) The content of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in SUDHL4 expressing control or CTPS1 sgRNAs detected by lipidomics analysis. (D) Relative PC and PE levels in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. (E) Schematic diagram of single-cell RNA sequencing (scRNA-seq) analysis based on the CTPS1 protein level. Representative immunohistochemical images of CTPS1 protein expression levels in DLBCL tumor tissues for single-cell RNA sequencing analysis. Brown signal was considered as positive staining. (F) Violin plot showing that the UCell scores for pathways associated with nucleotide metabolism, autophagy, and phospholipid metabolism were significantly elevated in the CTPS1 high expression group. (G, H) Gene Ontology (GO) enrichment (G) and gene set enrichment analysis (GSEA) (H) revealing that pathways related to nucleotide metabolism and phospholipid metabolism were significantly enriched in the CTPS1 high expression group. Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: Expressing, Control, Single Cell, RNA Sequencing, Immunohistochemical staining, Staining

Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: CTPS1 mediates phospholipid metabolic remodeling by enhancing CEPT1 in DLBCL. (A) The Venn diagram of the RNA-seq of SUDHL4 expressing control or CTPS1 sgRNAs, DepMap DLBCL essential gene and phospholipid metabolism gene. (B) Correlation analysis of CTPS1 and CEPT1 for DLBCL patients using the Gene Expression Profiling Interactive Analysis (GEPIA) database. (C) GEPIA database analysis of CEPT1 expression in pan-cancer. (D) Reads number analysis showing CEPT1 expression in SUDHL4 expressing control (n = 3) or CTPS1 sgRNAs (n = 3) by RNA-Seq. (E) CEPT1 mRNA level in SUDHL4 and SUDHL8 cells expressing control or CTPS1 sgRNAs. Levels in cells with sgControl were set to 1. (F) Immunoblot analysis showing the expression of CEPT1, CHPT1 in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs. n = 3. (G) Immunofluorescence analysis showing CEPT1 expression in SUDHL4 expressing control or CTPS1 sgRNAs. Scale bar: 20 μm (H) Immunohistochemical analysis of CTPS1 and CEPT1 expression in the subcutaneous tumor of SUDHL4 and SUDHL8 xenograft mouse model expressing control or CTPS1 sgRNAs. Brown signal in IHC was considered as positive staining. (I) Abundance analysis of CTP in SUDHL4 expressing control or CTPS1 sgRNAs by LC/MS. (J) CEPT1 mRNA level in SUDHL4 and SUDHL8 cells expressing control or CTPS1 sgRNAs, treated without or with CTP. Levels in cells with sgControl were set to 1. (K) Immunoblot analysis showing the expression of CEPT1 in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs, treated without or with CTP. (L, M) Relative PC and PE levels in SUDHL4 and SUDHL8 expressing control or CTPS1 sgRNAs, treated without or with CTP. Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: RNA Sequencing, Expressing, Control, Gene Expression, Western Blot, Immunofluorescence, Immunohistochemical staining, Staining, Liquid Chromatography with Mass Spectroscopy

Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: CTPS1 promotes BNIP3-mediated mitophagy through CEPT1. (A) Immunoblot analysis showing the knockout efficiency of sgCEPT1 and expression of PARP, cleaved PARP in SUDHL4 and SUDHL8 cells. (B) Growth curve analysis of SUDHL4 and SUDHL8 expressing control or CEPT1 sgRNAs expression. (C) Flow cytometry analysis showing cell apoptosis in SUDHL4 and SUDHL8 expressing control or CEPT1 sgRNAs using annexin V-APC/7-AAD Kit. (D) Immunoblot analysis of LC3 I/II expression in the mitochondria in SUDHL4 expressing control or CEPT1 sgRNAs without or with 20 nM Bafilomycin A1 (BafA1). (E) Immunoblot analysis of CTPS1, FLAG, LC3 I/II and BNIP3 expression in SUDHL4 and SUDHL8 expressing control, ovCTPS1, CEPT1 sgRNA, ovCTPS1 and CEPT1 sgRNA. (F, G) Relative PC and PE levels in SUDHL4 and SUDHL8 expressing control, ovCTPS1, CEPT1 sgRNA, ovCTPS1 and CEPT1 sgRNA. (H) The relative levels of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) detected by lipidomics in mitochondria immunoprecipitated from SUDHL4 cells expressing control or CEPT1 sgRNA. (I) Mitochondrial superoxide (MitoSOX) analysis in SUDHL4 and SUDHL8 expressing control or CEPT1 sgRNAs treated without or with PC and PE. (J) Mitochondrial membrane potential analysis in SUDHL4 and SUDHL8 expressing control or CEPT1 sgRNAs treated without or with PC and PE by JC-1. (K) MitoPerOx analysis measuring lipid peroxidation in SUDHL4 expressing control or CEPT1 sgRNAs treated without or with PC and PE. (L) Immunoblot analysis showing the expression of LC3 I/II, DRP1, p -DRP1S616, OPA1 and BNIP3 in SUDHL4 and SUDHL8 expressing control or CEPT1 sgRNAs treated without or with PC and PE. (M) Immunofluorescence analysis showing BNIP3 expression around mitochondria in SUDHL4 expressing control or CEPT1 sgRNAs treated without or with PC and PE. Scale bar: 20 μm. Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: Western Blot, Knock-Out, Expressing, Control, Flow Cytometry, Immunoprecipitation, Membrane, Immunofluorescence

Journal: Redox Biology

Article Title: CTPS1 modulates mitophagy to propel diffuse large B-cell lymphoma via reshaping CEPT1-mediated phospholipid metabolism

doi: 10.1016/j.redox.2026.104132

Figure Lengend Snippet: Selective CTPS1 inhibitor R80 reduces the viability of DLBCL cells. (A) CCK8 analysis showing relative cell viability of DLBCL cells after treated with R80 for 72h. (B) Immunoblot analysis showing cleaved PARP, BCL-XL expression in SUDHL4 and SUDHL8 treated with different concentrations of R80 for 72 h (C) Flow cytometry analysis showing cell apoptosis of SUDHL4 and SUDHL8 cells after treated with R80 for 24, 48, 72h using annexin V-APC/7-AAD Kit. (D) Immunoblot analysis showing CEPT1 expression in SUDHL4 and SUDHL8 treated with different concentrations R80 for 72 h (E, F) Relative PC and PE levels in SUDHL4 and SUDHL8 treated with different concentrations R80 for 72 h (G–I) Schematic diagram (N), volume analysis of tumor (H) and tumor growth curves analysis (I) of SUDHL8 xenograft mouse model treated with vehicle or R80 (n = 5 per group). R80 was dosed at 30 or 50 mg/kg days 1–4 of a 7-day cycle for 3 cycles. (J)Immunohistochemical analysis of CEPT1 expression in the subcutaneous tumor of SUDHL8 xenograft mouse model treated with vehicle or R80. Brown signal in IHC was considered as positive staining. (K, L) Relative PC and PE levels in the serum of SUDHL8 xenograft mouse model treated with vehicle or R80. (M)Schematic model of CTPS1 regulating BNIP3-mediated mitophagy by reshaping phospholipid metabolism in combination with CEPT1, resulting in the proliferation of DLBCL cells. Data are shown as the mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using one way ANOVA with multiple comparisons.

Article Snippet: DLBCL cell lines including FARAGE, SUDHL4, and SUDHL8 were obtained from the American Tissue Culture Collection (ATCC).

Techniques: Western Blot, Expressing, Flow Cytometry, Immunohistochemical staining, Staining