anti-pd1 antibody pembrolizumab  (Merck & Co)

Journal: Frontiers in Oncology

Article Title: Effects of Arylsulfatase B and Pembrolizumab in combination on progression of metastatic melanoma in the B16F10 syngeneic mouse model

doi: 10.3389/fonc.2026.1820206

Figure Lengend Snippet: Impact of ARSB and Pembrolizumab and their combination on pulmonary metastases in B16F10 melanoma. Thirty-two 10.5 week-old male C57BL/6J mice were inoculated with 250,000 B16F10 melanoma cells by tail vein injection. Two mice died of uncertain causes, and the remaining mice were euthanized on day 14 following tumor inoculation. Groups were: (A) untreated; (B) rhARSB (0.2 mg/kg IV on days 2, 7, and 12); (C) Pembrolizumab (5 mg/kg IP on days 2,7, and 12); and (D) combined rhARSB and Pembrolizumab treatments. (E) Compared to saline-treated control, the rhARSB, Pembrolizumab, and the combined rhARSB and Pembrolizumab groups had significantly fewer tumors (one-way ANOVA with Tukey post-test for multiple comparisons). Bar = 0.5 cm. (ARSB=arylsulfatase B=N-acetylgalactosamine-4-sulfatase; Pembro=Pembrolizumab rh=recombinant human).

Article Snippet: Pembrolizumab was purchased (HY-P9902, MedChemExpress, Mammoth Junction, NJ) and injected intraperitoneally at a concentration of 5 mg/kg.

Techniques: Injection, Saline, Control, Recombinant

Journal: Frontiers in Oncology

Article Title: Effects of Arylsulfatase B and Pembrolizumab in combination on progression of metastatic melanoma in the B16F10 syngeneic mouse model

doi: 10.3389/fonc.2026.1820206

Figure Lengend Snippet: Effects of Pembrolizumab, ARSB, and their combination on COP1-mediated pathway of apoptosis in B16F10 pulmonary melanomas and A375 human melanoma cells. (A) Cleaved caspase-3 increased following rhARSB or Pembrolizumab and was further Increased by their combination in treated B16F10 pulmonary melanomas. All p-values are calculated by one-way ANOVA with Tukey test for multiple comparisons. (B) In treated B16F10 melanomas, COP1 mRNA expression increased following rhARSB, but not following Pembrolizumab treatment. (C) BCL2 mRNA expression declined following either rhARSB or Pembrolizumab and declined further by their combination in the B16F10 pulmonary melanomas. (D) In cultured human A375 melanoma cells, cleaved caspase-3 increased following rhARSB and was further increased by the combination of rhARSB with activated PBMC [PBMC(Ac)]. Pembrolizumab alone or Pembrolizumab with unactivated PBMC did not increase cleaved caspase-3. However, cleaved caspase 3 was increased by the combination of Pembrolizumab with PBMC(Ac) and further increased by rhARSB, Pembrolizumab, and activated PBMC. (E) In the A375 cells, COP1 increased following rhARSB, but was unaffected by Pembrolizumab or by PBMC, either activated or not activated. (F) Inverse to the effect on COP1, rhARSB reduced the expression of BCL2. Neither Pembrolizumab alone nor PBMC alone affected BCL2 expression, but Pembro with activated PBMC reduced BCL2. BCL2 was further reduced by the combination of rhARSB + Pembrolizumab + PBMC(Ac) in the A375 cells. (G) Granzyme B increased following the combinations of rhARSB and PBMC(Ac) and Pembrolizumab and PBMC(Ac). Synergistic effect was evident following treatment by rhARSB + Pembrolizumab + and PBMC(Ac) in the A375 cells. (H) Viability was detected by the PrestoBlue assay in A375 cells. RhARSB, activated PBMC, and Pembrolizumab with activated PBMC reduced viability; decline was greatest by their combination. (I) Viability was also reduced in the B16F10 cells following rhARSB, Pembrolizumab with activated PBMC, and activated PBMC and most reduced by the combination. Declines were less than in the A375 cells. [Ac, activated; PBMC Ac, activated peripheral blood mononuclear cells; COP1, constitutive photomorphogenic protein 1; Pembro, Pembrolizumab; PBMC, peripheral blood mononuclear cells; rhARSB, recombinant human arylsulfatase B].

Article Snippet: Pembrolizumab was purchased (HY-P9902, MedChemExpress, Mammoth Junction, NJ) and injected intraperitoneally at a concentration of 5 mg/kg.

Techniques: Expressing, Cell Culture, Prestoblue Assay, Recombinant

Journal: Frontiers in Oncology

Article Title: Effects of Arylsulfatase B and Pembrolizumab in combination on progression of metastatic melanoma in the B16F10 syngeneic mouse model

doi: 10.3389/fonc.2026.1820206

Figure Lengend Snippet: Matrix metalloproteinases following rhARSB and Pembrolizumab in B16F10 pulmonary metastatic melanomas and in A375 cells. (A, B) mRNA expression of MMP9 and MMP2 declined following rhARSB and was unchanged by Pembrolizumab in the pulmonary melanomas. (C) Serum MMP2 declined more following rhARSB than following Pembrolizumab and was further reduced by combined treatment in the metastatic pulmonary melanomas. (D) Total MMP activity in spent media of A375 cells declined following rhARSB and increased following ARSB siRNA, but was unaffected by Pembrolizumab. (E, F) MMP9 and MMP29 mRNA expression in A375 cells was unaffected by Pembrolizumab, either alone or in combination with unactivated or activated PBMC. In contrast, rhARSB significantly reduced their expression. (G) Migration of A375 cells declined following rhARSB, increased following ARSB silencing, and was unaffected by Pembrolizumab. [Ac, activated; ARSB, arylsulfatase B, N-acetylgalactosamine-4-sulfatase; MMP-matrix, metalloproteinase; PBMC, peripheral blood mononuclear cells; Pembro, Pembrolizumab; rh, recombinant human; si, siRNA].

Article Snippet: Pembrolizumab was purchased (HY-P9902, MedChemExpress, Mammoth Junction, NJ) and injected intraperitoneally at a concentration of 5 mg/kg.

Techniques: Expressing, Activity Assay, Migration, Recombinant

Journal: Frontiers in Oncology

Article Title: Effects of Arylsulfatase B and Pembrolizumab in combination on progression of metastatic melanoma in the B16F10 syngeneic mouse model

doi: 10.3389/fonc.2026.1820206

Figure Lengend Snippet: Expression of cytokines/chemokines in B16F10 melanoma lung tissue following rhARSB and Pembrolizumab. (A, B) In B16F10 pulmonary melanoma tissue, mRNA expression of MCP1 and IL-10 increased following treatment by Pembrolizumab or rhARSB, and the increase was enhanced by their combination. (C) In contrast, IL-17α expression was increased by Pembrolizumab, but not by rhARSB. (D, E) KC, the mouse analog of IL-8, and IL-6 were reduced by both rhARSB and Pembrolizumab and further reduced by their combination. (F) In contrast, TNFα was increased by Pembrolizumab and reduced by rhARSB, with overall reduction by their combination. (G, H, I) FGF2, VEGF, and EGF were reduced by ARSB, but not by Pembrolizumab, and the combination of ARSB and Pembrolizumab did not lead to additional decline. [ARSB, arylsulfatase B, N-acetylgalactosamine-4-sulfatase; EGF, epidermal growth factor; bFGF, basic fibroblast growth factor; IL, interleukin; KC CXCL1, keratinocyte-derived cytokine; rh, recombinant human; MCP1 CCL2, monocyte chemoattractant protein-1; Pembro, Pembrolizumab; VEGF, vascular endothelial growth factor].

Article Snippet: Pembrolizumab was purchased (HY-P9902, MedChemExpress, Mammoth Junction, NJ) and injected intraperitoneally at a concentration of 5 mg/kg.

Techniques: Expressing, Derivative Assay, Recombinant

Journal: Frontiers in Oncology

Article Title: Effects of Arylsulfatase B and Pembrolizumab in combination on progression of metastatic melanoma in the B16F10 syngeneic mouse model

doi: 10.3389/fonc.2026.1820206

Figure Lengend Snippet: Effects of rhARSB and Pembrolizumab on polymorphonuclear (PMN) leukocyte invasion and apoptosis in A375 cells. (A) PMN invasion to the A375 cells was increased by rhARSB, reduced by ARSB siRNA, reduced by IL-8 silencing and increased by addition of rhIL-8. These effects are attributable to changes in IL-8 in the spent media. PMN migration from the filter on top to the bottom well was increased by rhIL-8 in the bottom well, but unaffected by Pembrolizumab. (C) IL-8 in the spent media of A375 cells following rhARSB increased and declined when ARSB was silenced. (D) In contrast, IL-8 in the cell extract declined following rhARSB and increased following ARSB siRNA. (E) Total IL-8 in the A375 cells declined following rhARSB and increased following ARSB knockdown. (F) When IL-8 was silenced in the A375 cells, the effect of rhARSB + PBMC(Ac) and of Pembro + PBMC(Ac) and their combination [rhARSB+Pembro+PBMC(Ac)] on cleaved caspase-3 was reduced, consistent with an effect of IL-8 on PBMC(Ac)-mediated apoptosis. (G) When PMN were added to the combination of rhARSB+Pembrolizumab+PBMC, the cleaved caspase-3 was further increased in the A375 cells. [Ac, activated; consi, control siRNA; IL, interleukin; rh, recombinant human; N.D., no difference; Pembro, Pembrolizumab; PBMC, peripheral blood mononuclear cells; PMN, polymorphonuclear leukocytes; si, siRNA].

Article Snippet: Pembrolizumab was purchased (HY-P9902, MedChemExpress, Mammoth Junction, NJ) and injected intraperitoneally at a concentration of 5 mg/kg.

Techniques: Migration, Knockdown, Control, Recombinant

Journal: iScience

Article Title: Decoding drug-responsive cell subpopulations in triple-negative breast cancer using single-cell multiomics

doi: 10.1016/j.isci.2026.115445

Figure Lengend Snippet: Decoding drug response in patient samples using single-cell identity annotation (A) UMAP plots show cluster identifications, cell types, and treatment conditions for all single cells in the BIOKEY_9 (B9) tumor. These plots provide an overview of the cellular landscape within the tumor, highlighting distinct clusters and their associations with treatment conditions. (B) Single-cell identity changes in the cancer cells of TNBC patient BIOKEY_9 (B9) after pembrolizumab treatment. Pie charts illustrate the distribution of single cells across different identities within the cancer cell population before and after treatment. (C) Identity scores for single cells in the BIOKEY_9 (B9) cancer cell population before and after pembrolizumab treatment. These scores quantify the intensity of gene expression identities at the single-cell level, revealing shifts in cellular identities induced by treatment. Boxplot center line indicates median; box bounds represent the interquartile range (IQR); whiskers extend to 1.5×IQR. (D) GSEA results for F1 and F2 gene sets after pembrolizumab treatment, highlighting the enrichment or depletion of these identity-specific gene sets in the treated cancer cell population. (E) Expression scores for three MsigDB hallmark gene sets in BIOKEY_9 (B9) cancer single cells before and after treatment. The hallmark gene sets analyzed include estrogen response (early and late combined), TNFα signaling via NF-κB, and epithelial-mesenchymal transition (EMT), providing insights into treatment-induced changes in key biological pathways. Violin plots show the distribution of expression scores. (F) Volcano plots of differentially expressed genes (DEGs) in BIOKEY_9 (B9) cancer single cells after pembrolizumab treatment. Pie charts accompanying the plots display the percentage of DEGs associated with each identity in the CCLE model. The right pie chart represents upregulated genes, while the left represents downregulated genes, linking these changes to specific identities. (G) Analysis of identity composition changes in three cancer cell clusters from BIOKEY_9, B9 (cluster_0, cluster_6, and cluster_3), before and after pembrolizumab treatment. Pie charts illustrate the percentage of single cells belonging to each identity within each cluster, revealing differential responses across clusters. (H) Global characterization of identity enrichment or depletion for cancer cells within 11 TNBC tumors following anti-PD1 treatment. Chi-squared test results assess the significance of identity changes across tumors, with stacked bar plots depicting residual scores for each identity. Positive residual scores indicate identity expansion, while negative scores indicate depletion. (I) Analysis of cancer cell clusters contributing to the largest identity expansions in cancer cells for each tumor. Chi-squared test p -values assess whether specific clusters are more associated with identity expansion compared to other clusters within the same tumor. Stacked bar plots represent the residual scores for each cluster, highlighting their contributions to identity changes. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by the Wilcoxon rank-sum test (C and E) or by chi-squared test (H and I).

Article Snippet: As a proof-of-concept, we applied our approach to a representative patient sample (BIOKEY_9) with tumor samples collected before and after pembrolizumab monotherapy ( A).

Techniques: Single Cell, Gene Expression, Expressing

Journal: iScience

Article Title: Decoding drug-responsive cell subpopulations in triple-negative breast cancer using single-cell multiomics

doi: 10.1016/j.isci.2026.115445

Figure Lengend Snippet: Decoding drug response in patient samples using single-cell identity annotation (A) UMAP plots show cluster identifications, cell types, and treatment conditions for all single cells in the BIOKEY_9 (B9) tumor. These plots provide an overview of the cellular landscape within the tumor, highlighting distinct clusters and their associations with treatment conditions. (B) Single-cell identity changes in the cancer cells of TNBC patient BIOKEY_9 (B9) after pembrolizumab treatment. Pie charts illustrate the distribution of single cells across different identities within the cancer cell population before and after treatment. (C) Identity scores for single cells in the BIOKEY_9 (B9) cancer cell population before and after pembrolizumab treatment. These scores quantify the intensity of gene expression identities at the single-cell level, revealing shifts in cellular identities induced by treatment. Boxplot center line indicates median; box bounds represent the interquartile range (IQR); whiskers extend to 1.5×IQR. (D) GSEA results for F1 and F2 gene sets after pembrolizumab treatment, highlighting the enrichment or depletion of these identity-specific gene sets in the treated cancer cell population. (E) Expression scores for three MsigDB hallmark gene sets in BIOKEY_9 (B9) cancer single cells before and after treatment. The hallmark gene sets analyzed include estrogen response (early and late combined), TNFα signaling via NF-κB, and epithelial-mesenchymal transition (EMT), providing insights into treatment-induced changes in key biological pathways. Violin plots show the distribution of expression scores. (F) Volcano plots of differentially expressed genes (DEGs) in BIOKEY_9 (B9) cancer single cells after pembrolizumab treatment. Pie charts accompanying the plots display the percentage of DEGs associated with each identity in the CCLE model. The right pie chart represents upregulated genes, while the left represents downregulated genes, linking these changes to specific identities. (G) Analysis of identity composition changes in three cancer cell clusters from BIOKEY_9, B9 (cluster_0, cluster_6, and cluster_3), before and after pembrolizumab treatment. Pie charts illustrate the percentage of single cells belonging to each identity within each cluster, revealing differential responses across clusters. (H) Global characterization of identity enrichment or depletion for cancer cells within 11 TNBC tumors following anti-PD1 treatment. Chi-squared test results assess the significance of identity changes across tumors, with stacked bar plots depicting residual scores for each identity. Positive residual scores indicate identity expansion, while negative scores indicate depletion. (I) Analysis of cancer cell clusters contributing to the largest identity expansions in cancer cells for each tumor. Chi-squared test p -values assess whether specific clusters are more associated with identity expansion compared to other clusters within the same tumor. Stacked bar plots represent the residual scores for each cluster, highlighting their contributions to identity changes. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by the Wilcoxon rank-sum test (C and E) or by chi-squared test (H and I).

Article Snippet: Violin plots show the distribution of expression scores. (F) Volcano plots of differentially expressed genes (DEGs) in BIOKEY_9 (B9) cancer single cells after pembrolizumab treatment.

Techniques: Single Cell, Gene Expression, Expressing