Journal: Human Mutation

Article Title: Combined Nivolumab and Ipilimumab Therapy Promotes Immune‐Mediated Cardiomyocyte Apoptosis Through TLR4–Myd88–NF‐ Κ b–Driven Activation of the NLRP3 Inflammasome

doi: 10.1155/humu/8506248

Figure Lengend Snippet: Nivolumab combined with ipilimumab activates the TLR4–MyD88–NF‐ κ B pathway, whereas TLR4 or NLRP3 knockdown attenuates apoptosis‐related changes in AC16 cardiomyocytes. (A) The expression of TLR4–MyD88–NF‐ κ B pathway‐related proteins was detected via Western blot. (B–G) The protein expression levels of TLR4, MyD88, NF‐ κ B, p‐NF‐ κ B, IKK β , and p‐IKK β . Single‐drug group: PD‐1, programmed cell death Protein 1; combination drug group: CTLA‐4, cytotoxic T lymphocyte‐associated antigen‐4; NF‐ κ B, nuclear factor kappa‐B; TLR4, Toll‐like Receptor 4. (H) Representative Western blot showing the effects of TLR4 or NLRP3 silencing on apoptosis‐related protein expression in AC16 cardiomyocytes treated with nivolumab combined with ipilimumab. (I–M) Quantification of apoptosis‐related proteins, including NLRP3, Bcl‐2, BAX, cleaved caspase‐3, and caspase‐3. (N) Representative Western blot showing the effects of TLR4 silencing on NLRP3 inflammasome components and TLR4–MyD88–NF‐ κ B pathway‐related proteins. (O–U) Quantification of NLRP3, TLR4, MyD88, NF‐ κ B, p‐NF‐ κ B, IKK β , and p‐IKK β expression levels. Data are presented as mean ± SD from three independent biological replicates ( n = 3). Statistical significance was determined using one‐way ANOVA followed by Dunnett′s post hoc test for multiple comparisons. p < 0.05; ∗ p < 0.01; ∗∗ p < 0.001; ns, not significant.

Article Snippet: The nivolumab (anti‐PD‐1) PD‐1 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9903; the ipilimumab (anti‐CTLA‐4) CTLA‐4 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9901; the si‐TLR4 inhibitor was purchased from Guangzhou Ruibo; and the si‐NLRP3 inhibitor was purchased from Guangzhou Ruibo.

Techniques: Knockdown, Expressing, Western Blot

Journal: Human Mutation

Article Title: Combined Nivolumab and Ipilimumab Therapy Promotes Immune‐Mediated Cardiomyocyte Apoptosis Through TLR4–Myd88–NF‐ Κ b–Driven Activation of the NLRP3 Inflammasome

doi: 10.1155/humu/8506248

Figure Lengend Snippet: Nivolumab combined with ipilimumab enhanced the degree of apoptosis and inflammation in mouse cardiomyocytes. (A, B) The expression of the myocardial injury factor BNP was detected via immunofluorescence staining. (C) The expression of the myocardial injury factors BNP and TnT was detected by immunohistochemistry. (D–I) The expression of the cardiac apoptosis‐related proteins BAX, Bcl‐2, cleaved caspase‐3, and caspase‐3 was detected via immunofluorescence staining and Western blot. Single group: PD‐1, programmed cell death Protein 1; combination group: CTLA‐4, cytotoxic T lymphocyte‐associated antigen‐4. Data are presented as mean ± SD. Each group consisted of five biological replicates (mice) ( n = 5). Statistical significance was determined using one‐way ANOVA followed by Dunnett′s post hoc test for multiple comparisons. p < 0.05; ∗ p < 0.01; ∗∗ p < 0.001; ns, not significant.

Article Snippet: The nivolumab (anti‐PD‐1) PD‐1 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9903; the ipilimumab (anti‐CTLA‐4) CTLA‐4 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9901; the si‐TLR4 inhibitor was purchased from Guangzhou Ruibo; and the si‐NLRP3 inhibitor was purchased from Guangzhou Ruibo.

Techniques: Expressing, Immunofluorescence, Staining, Immunohistochemistry, Western Blot

Journal: Human Mutation

Article Title: Combined Nivolumab and Ipilimumab Therapy Promotes Immune‐Mediated Cardiomyocyte Apoptosis Through TLR4–Myd88–NF‐ Κ b–Driven Activation of the NLRP3 Inflammasome

doi: 10.1155/humu/8506248

Figure Lengend Snippet: Nivolumab combined with ipilimumab enhances the expression of the TLR4–Myd88–NF‐ κ B signaling pathway and NLRP3 in the apoptosis of mouse cardiomyocytes. (A) Western blot analysis of the expression of inflammatory factors in the myocardial tissue of mice in the single drug group and combined drug group. (B–F) The expression levels of NLRP3, ASC, caspase‐1, IL‐18 and IL‐1 β . (G) Western blot analysis of TLR4–MyD88–NF‐ κ B pathway‐related proteins in the myocardial tissue of mice in the single‐drug group and (H–M) combination drug group, including the protein expression levels of TLR4, Myd88, NF‐ κ B, p‐NF‐ κ B, IKK β , and p‐IKK β . Single‐drug group: PD‐1, programmed cell death Protein 1. The combination drugs used were as follows: CTLA‐4, cytotoxic T lymphocyte–associated antigen‐4; NF‐ κ B, nuclear factor kappa‐B; TLR4, Toll‐like Receptor 4; and NLRP3, NOD‐like receptor thermal protein domain associated Protein 3. Data are presented as mean ± SD. Each group consisted of five biological replicates (mice) ( n = 5). Statistical significance was determined using one‐way ANOVA followed by Dunnett’s post hoc test for multiple comparisons. p < 0.05; ∗ p < 0.01; ∗∗ p < 0.001; ns, not significant.

Article Snippet: The nivolumab (anti‐PD‐1) PD‐1 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9903; the ipilimumab (anti‐CTLA‐4) CTLA‐4 inhibitor was purchased from MCE (United States) Biotechnology Company, Product Number HY‐P9901; the si‐TLR4 inhibitor was purchased from Guangzhou Ruibo; and the si‐NLRP3 inhibitor was purchased from Guangzhou Ruibo.

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Article Snippet: Immune checkpoint inhibitors (ICIs) used in this study: anti-CTLA-4 (BioXCell cat. #BE0131) and anti-PD-1 (BioXCell cat. #BE0146).

Techniques: Single Cell, RNA Sequencing, Expressing, Isolation, Derivative Assay, Fluorescence