Journal: Communications Biology

Article Title: Noncanonical amino acid-aided synthesis of anti-PD-L1 bispecific nanobody for colon cancer immunotherapy

doi: 10.1038/s42003-025-09222-1

Figure Lengend Snippet: A Experimental design for in vivo tumor-challenge and PV2-S71 + PV3-N77 treatment plan. MC38 xenograft mice model was established. PV2-PV3, PV2-S71 + PV3-N77 and durvalumab (10 mg/kg, once every three days for six times) was treated for 15 days when the tumor grew to 80–100 mm 3 . (created with BioRender.com, with permission); B The tumor volume of mice treated with different treatment strategies; C Body weight of each group ( n = 6 biologically independent samples); D The tumor weight and image of tumors isolated from tumor-bearing mice on day 18; E Effect of bispecific nanobodies on the cytokine concentration in serum; F H&E and IHC staining of tumor tissues; G The expression of Ki67 and CD8 were confirmed using an immunohistochemistry assay in tumor tissues. (x ± SD, n = 6 biologically independent samples; a significant difference from the Normal saline group, *: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: The human PD-L1 MC38 CRC cell line was obtained from Shanghai Model Organisms Center, Inc. (Shanghai, China). hPD-L1 MC38 cells were cultivated in Dulbecco’s Modified Eagle Medium containing 10% (v/v) fetal bovine serum (FBS), 50 μg/mL hygromycin B, and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin).

Techniques: In Vivo, Isolation, Concentration Assay, Immunohistochemistry, Expressing, Saline

Journal: Communications Biology

Article Title: Noncanonical amino acid-aided synthesis of anti-PD-L1 bispecific nanobody for colon cancer immunotherapy

doi: 10.1038/s42003-025-09222-1

Figure Lengend Snippet: A Single-cell suspensions from the excised MC38 tumors treated as indicated were stained for immune cell markers CD45, CD3, CD4, and CD8 for T cells according to the analysis method shown in ( B ); C CD8 + IFN-γ + T cell were stained and analyzed; D The CD8 + T cells infiltration in each group; E The ratio of CD8 + IFN-γ + T cells in the tumor. (x ± SD, n = 6 biologically independent samples; a significant difference from the Normal saline group, *: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: The human PD-L1 MC38 CRC cell line was obtained from Shanghai Model Organisms Center, Inc. (Shanghai, China). hPD-L1 MC38 cells were cultivated in Dulbecco’s Modified Eagle Medium containing 10% (v/v) fetal bovine serum (FBS), 50 μg/mL hygromycin B, and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin).

Techniques: Staining, Saline

Journal: Oncology Reports

Article Title: Continuous administration of bevacizumab plus capecitabine, even after acquired resistance to bevacizumab, restored anti-angiogenic and antitumor effect in a human colorectal cancer xenograft model

doi: 10.3892/or.2016.4902

Figure Lengend Snippet: Bevacizumab shows antitumor activity in the early phase but not in the late phase in HT-29 xenograft model. Mice were randomized into 2 groups (n=5–10/group). Bevacizumab and control human IgG were intraperitoneally administered once a week. Data points show mean ± SD of tumor volume. Control, open circles; bevacizumab, closed circles. a P<0.05 versus control IgG group by Wilcoxon test.

Article Snippet: HT-29 tumor tissues collected on day 50 from the bevacizumab PD model were homogenized with cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA) with protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA) and phosphatase inhibitor cocktail (Nacalai Tesque, Kyoto, Japan).

Techniques: Activity Assay, Control

Journal: Oncology Reports

Article Title: Continuous administration of bevacizumab plus capecitabine, even after acquired resistance to bevacizumab, restored anti-angiogenic and antitumor effect in a human colorectal cancer xenograft model

doi: 10.3892/or.2016.4902

Figure Lengend Snippet: Combination therapy with bevacizumab plus capecitabine inhibits tumor growth and tumor angiogenesis in the HT-29 xenograft bevacizumab PD model. Antitumor activity (A) and anti-angiogenic activity (B) of combination therapy with capecitabine plus bevacizumab in the HT-29 xenograft model unresponsive to bevacizumab. Mice that had been treated with bevacizumab on days 1 and 8 were randomly allocated to control, bevacizumab, capecitabine, and bevacizumab plus capecitabine groups on day 29 (n=7/group). CD31 immunostaining in tumor tissue at day 50 (n=7/group). Data points show mean ± SD of tumor volume. Control, open circles; bevacizumab, closed circles; capecitabine, cross marks; combination group, closed triangles. The box-and-whisker plots show mean ± SD. a P<0.05 versus control group; b P<0.05 versus capecitabine group; c P<0.05 versus bevacizumab group by Wilcoxon test.

Article Snippet: HT-29 tumor tissues collected on day 50 from the bevacizumab PD model were homogenized with cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA) with protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA) and phosphatase inhibitor cocktail (Nacalai Tesque, Kyoto, Japan).

Techniques: Activity Assay, Control, Immunostaining, Whisker Assay

Journal: Oncology Reports

Article Title: Continuous administration of bevacizumab plus capecitabine, even after acquired resistance to bevacizumab, restored anti-angiogenic and antitumor effect in a human colorectal cancer xenograft model

doi: 10.3892/or.2016.4902

Figure Lengend Snippet: Combination therapy did not change the expression of stromal cell-derived angiogenic factors in tumor tissues. Changes in stromal cell-derived angiogenic factors. Membranes-based antibody arrays were reacted with homogenized tumor tissue harvested from the control, bevacizumab, capecitabine, and combination group on day 50.

Article Snippet: HT-29 tumor tissues collected on day 50 from the bevacizumab PD model were homogenized with cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA) with protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA) and phosphatase inhibitor cocktail (Nacalai Tesque, Kyoto, Japan).

Techniques: Expressing, Derivative Assay, Control

Journal: Oncology Reports

Article Title: Continuous administration of bevacizumab plus capecitabine, even after acquired resistance to bevacizumab, restored anti-angiogenic and antitumor effect in a human colorectal cancer xenograft model

doi: 10.3892/or.2016.4902

Figure Lengend Snippet: Combination therapy suppresses tumor cell-derived VEGF and galectin-3 in tumor tissues. (A) Membranes-based antibody arrays were reacted with tumor tissue homogenates from the control group, bevacizumab group, capecitabine group, and combination group on day 50. (B) Levels of free VEGF in the tumor tissues (n=7). (C) Levels of galectin-1 and galectin-3 in the tumor tissues (n=7). The control group (open), bevacizumab group (horizontal striped), capecitabine group (hatched), and combination group (closed). Columns show mean ± SD. a P<0.05 versus control by Wilcoxon test. (D) Downregulation of galectin-3 expression by 5-FU in in vitro culture. Culture supernatant was collected and subjected to ELISA (n=3). Columns show mean ± SD. a P<0.05 versus control by Dunnett's test.

Article Snippet: HT-29 tumor tissues collected on day 50 from the bevacizumab PD model were homogenized with cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA) with protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA) and phosphatase inhibitor cocktail (Nacalai Tesque, Kyoto, Japan).

Techniques: Derivative Assay, Control, Expressing, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) Immunoblot of PD-1 expression in human TNBC tumor tissues. Black circle, glycosylated PD-1; arrowhead, non-glycosylated PD-1. (B) Immunoblot of PD-1 expression in shCTRL, shPD-1, and PD-1-re-expressing Jurkat T cells stimulated by PHA overnight. (C) Glycoprotein staining and Coomassie blue staining of PNGase F-treated purified PD-1. Horseradish peroxidase (HRP) and soybean trypsin inhibitor (STI) served as positive and negative control, respectively. (D) Immunoblot of PD-1 in PD-1-expressing Jurkat (Jurkat-PD-1) T cells treated with inhibitors blocking N-linked or O-linked glycosylation as indicated. (E) Schematic diagram of PD-1 amino acid sequence alignment among different species. The four putative NXT motifs are shown in red. (F) Schematic diagram of full-length PD-1. ECD, extracellular domain; ICD, intracellular domain; SP, signal peptide; TM, transmembrane domain. Four putative NXT motifs in the ECD domain are labeled in red. The numbers indicate the amino acid positions. (G) Immunoblot of the protein expression pattern of PD-1 WT and indicated NQ mutants overexpressed in Jurkat T cells.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Western Blot, Expressing, Staining, Purification, Negative Control, Blocking Assay, Sequencing, Labeling

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) LC-MS/MS-based analysis of N-glycopeptides containing N74-glycans. PD-1 was purified from Jurkat-PD-1 FLAG cells treated with or without PHA overnight. The cartoon symbols used for the glycans conform to the standard representation recommended by the Consortium for Functional Glycomics. (B) Immunoblot of PD-1 in Jurkat T cells treated with PHA for the indicated time. (C) Lectin immunoblot of PD-1 purified from Jurkat-PD-1 FLAG treated with or without PHA for 5 hours. (D) Quantification of mRNA levels of various glycosyltransferases by real-time PCR in Jurkat T cells stimulated with α−CD3/CD28 for the indicated time points. All data represent mean ± SD from at least three independent experiments. *, P < 0.05.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Liquid Chromatography with Mass Spectroscopy, Purification, Functional Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) Schematic diagram of various PD-1 NQ mutants. The numbers indicate the amino acid positions on PD-1. (B) Flow cytometric analysis of cell surface PD-1 WT or the indicated NQ mutants over-expressed in Jurkat T cells. (C) Protein half-life of PD-1 WT or the indicated NQ mutants over-expressed in Jurkat T cells. Cells were treated with CHX for the indicated time, and PD-1 levels were examined by immunoblotting. (D) Protein half-life of PD-1 WT or 4NQ overexpressed in 293T cells. Cells were treated with CHX for the indicated time and PD-1 levels were examined by immunoblotting. (E) Ubiquitination of PD-1 WT or 4NQ purified from 293T cells treated with or without MG132. (F) Immunoblot of PD-1 4NQ in 293T cells treated with CHX for the indicated time in the presence or absence of MG132.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Western Blot, Purification

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) In vitro plate-based binding analysis of purified PD-1 with PD-L1 Fc in the absence or presence of PNGase F. (B) Analysis of PD-1/PD-L1 binding by immunoprecipitation. The lysates of 293T cells overexpressing FLAG-tagged PD-1 WT or 4NQ mutant were incubated with PD-L1 Fc and anti-FLAG M2 agarose, and then subjected to IP-Western. (C) Time-lapse microscopy of the dynamic interaction between green fluorescent-labeled PD-L1-Fc and PD-1 WT or 4NQ expressed in 293T cells. (D) Time-lapse evaluation and quantitation of the dynamic interaction between green fluorescent-labeled PD-L1-Fc and PD-1 WT or the indicated mutants expressed in Jurkat cells. (E) Mapping of the glycosylated sites critical for binding with PD-L1 by immunoprecipitation. The lysates of Jurkat cells overexpressing PD-1 WT or the indicated NQ mutants were incubated with pan mouse IgG dynabeads and Fc (mouse IgG) fused human PD-L1, and then subjected to IP-Western. The band intensity was quantified and normalized to show PD-L1 binding levels (right panel). (F) Flow cytometric analysis of the binding of PD-L1 Fc with cell surface PD-1 by using Jurkat T cells expressing PD-1 WT or the indicated NQ mutants. Median Fluorescence Intensity was quantified and normalized to show PD-L1 binding levels (right panel). All data represent mean ± SD from at least three independent experiments. *, P < 0.05.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: In Vitro, Binding Assay, Purification, Immunoprecipitation, Mutagenesis, Incubation, Western Blot, Time-lapse Microscopy, Labeling, Quantitation Assay, Expressing, Fluorescence

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) Dot blot analysis of PD-1 antibodies using purified PD-1 treated with or without PNGase F. (B) Schematic diagram of the live-cell imaging PD-1/PD-L1 binding assay in the presence of gPD-1 antibody. (C) Measurement of the neutralizing activity of gPD-1 antibodies using an IncuCyte system. 293T-PD-1 cells were incubated with fluorescent-labeled PD-L1-Fc fusion protein in the presence of gPD-1 antibodies. The binding of PD-1 and PD-L1 was quantified by counting fluorescent objects per mm2 using the Incucyte™ Zoom system every 2 hours. (D) EC50 calculation of STM418 by GraphPad Prism software. Experiments were conducted as described in (C) in the presence of indicated doses of STM418. (E) KD determination of STM418 or nivolumab. (F) Glyco-specificity evaluation of STM418 by immunoblot analysis of PD-1 WT or the indicated NQ mutants over-expressed in 293T cells. (G) Time-lapse evaluation and quantitation of the dynamic interaction between green fluorescent-labeled PD-L1-Fc and PD-1 WT or the indicated mutants expressed in Jurkat cells in the absence of presence of STM418. PD-L1 binding inhibition is represented by the fluorescence intensity ratio of the STM418 to IgG treatment in each cell line. (H) Epitope mapping of STM418 by high-mass MALDI mass spectrometry (CovalX). Sequences on the PD-1 protein shown in red represent the STM418 antibody binding sites.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Dot Blot, Purification, Live Cell Imaging, Binding Assay, Activity Assay, Incubation, Labeling, Software, Western Blot, Quantitation Assay, Inhibition, Fluorescence, Mass Spectrometry

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) Quantification of the proliferation of NCI-H226 cells co-cultured with T cells isolated from PMBCs in the presence or absence of STM418 (20 μg/mL). (B) Flow cytometric analysis of the proliferation of CFSE-labeled T cells co-cultured with dendritic cells (DCs) in the presence or absence of STM418 (10 μg/mL). (C) Quantification of IL-2 levels by ELISA. Experiments were conducted as described in (B). Supernatants were collected on day 5 and subjected to ELISA. (D) Normalized luminescence of PD-1 expressing Jurkat T cells transiently transfected with an NFAT-Luc reporter construct and stimulated with α-CD3/CD28/IgG or α-CD3/CD28/PD-L1 in the presence or absence of STM418 (20 μg/mL). (E) Flow cytometric analysis of the proliferation of CFSE-labeled T cells co-cultured with dendritic cells (DCs) in the presence of the indicated dose of IgG, STM418, nivolumab, or pembrolizumab. (F) Quantification of IFNγ levels by ELISA. Experiments were conducted as described in (E). Supernatants were collected on day 5 and subjected to ELISA. All data represent mean ± SD from at least three independent experiments. *, P < 0.05.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Cell Culture, Isolation, Labeling, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, Construct

Journal: Cancer research

Article Title: Targeting glycosylated PD-1 induces potent anti-tumor immunity

doi: 10.1158/0008-5472.CAN-19-3133

Figure Lengend Snippet: (A) Schematic diagram of a drug intervention protocol for anti-PD-1 antibodies (5 mg/kg) in a humanized animal model. (B) The growth of orthotopic MDA-MB-231 tumors in PD-1 antibody-treated SCID-PBMC mice. Tumors were measured until tumors reached > 100 mm3 in volume at the indicated time points. Nivo, nivolumab; Pembro, pembrolizumab. (C) The weight of tumors at the drug intervention endpoint. (D) Intracellular cytokine staining of CD8+ and granzyme B+ cells in CD3+ T-cell populations from isolated tumor-infiltrating lymphocytes. Results are presented as the mean ± standard deviation from a representative experiment. (E) Kaplan Meier survival curve for mice bearing MDA-MB-231 tumors after the drug intervention with anti-PD-1 antibodies. Significance was determined using the log-rank test. (F) Quantitative analysis of indicated biochemistry indices for liver and kidney function after the experiments. All error bars represent mean ± SD. *P < 0.05; NS, not significant.

Article Snippet: Protein were eluted in 2× SDS-sample buffer and subjected to Western blot with rabbit anti-PD-1 antibody (Cell Signaling Technology).

Techniques: Animal Model, Staining, Isolation, Standard Deviation

Journal: Science advances

Article Title: Small-molecule PIK-93 modulates the tumor microenvironment to improve immune checkpoint blockade response.

doi: 10.1126/sciadv.ade9944

Figure Lengend Snippet: Fig. 3. PIK-93 induces CUL4A-mediated proteasomal degradation of PD-L1. (A) PIK-93 reduces the half-life of endogenous PD-L1. CL83 cells were treated with cy- cloheximide (CHX) for the indicated time points with or without 10 μM PIK-93. In- dicated proteins were examined by im- munoblotting. (B) MG132 reverses the effects of PIK-93 on PD-L1. CL83 cells were treated with 0, 1, or 10 μM PIK-93 for 18 hours with or without 10 μM MG132 for another 6 hours. Indicated proteins were measured by immunoblotting. (C) PIK-93 induces ubiquitination of PD-L1. Human embryonic kidney (HEK) 293T cells were transiently transfected with PD-L1–Flag and incubated in the absence or presence of PIK-93 and MG132 (10 μM). After 6 hours, the cells were harvested, and PD- L1 was immunoprecipitated with an anti–PD-L1 antibody. Ubiquitination of PD-L1 was detected by immunoblotting. Immunoglobulin G (IgG) served as the antibody negative control for the immu- noprecipitation (IP) experiment. Ub, ubiq- uitin. (D) MLN4924 restores PD-L1 protein levels. Cells were treated with DMSO or PIK-93 for 24 hours and then cotreated with different doses of MLN4924 for 6 hours. Cdt served as MLN4924-positive control. Indicated proteins were measured by immunoblotting. (E) CUL knockdown effect on PIK-93–induced PD-L1 polyubi- quitination. CL83 cells transduced with CUL1–short hairpin RNA (shRNA), CUL4A- shRNA, or shLacZ control lentiviruses were incubated with or without PIK-93. Indi- cated proteins were detected by immu- noblotting. (F) PD-L1 and CUL4A form an interaction complex. PD-L1–Flag and hemagglutinin (HA)–CUL4A were overex- pressed in HEK293T cells. Reciprocal co-IP was conducted with an anti-Flag antibody (top panel) or an anti-HA antibody (bottom panel) followed by immunoblot- ting. (G) PIK-93 induces the interaction of PD-L1 and Cullin-4A. HEK293T cells were transiently transfected with PD-L1–Flag and then incubated with or without PIK- 93 and MG132 (10 μM). After 6 hours of treatment, the cells were harvested, and PD-L1 was immunoprecipitated with an anti–PD-L1 antibody, followed by immunoblotting.

Article Snippet: On the same day, the mice were injected intraperitoneally with DMSO control + human IgG1 antibody (HG1K, Sino Biological), DMSO control + anti–PD-L1 antibody (10mg/kg; atezolizumab, MedChemExpress), PIK-93 (60 mg/kg) + human IgG1 antibody, or the combination of PIK-93 (60 mg/kg) + anti–PD-L1 antibody (10 mg/kg) once every other day for 2 weeks.

Techniques: Western Blot, Ubiquitin Proteomics, Transfection, Incubation, Immunoprecipitation, Negative Control, Positive Control, Knockdown, Transduction, shRNA, Control, Co-Immunoprecipitation Assay

Journal: Science advances

Article Title: Small-molecule PIK-93 modulates the tumor microenvironment to improve immune checkpoint blockade response.

doi: 10.1126/sciadv.ade9944

Figure Lengend Snippet: Fig. 6. Combined treatment with PIK-93 and anti-PDL1 antibodies induces T cell activity, increases TIL recruitment, and enhances PD-L1 blockade efficacy in syn- geneic mouse tumor model and human PBMC CDX mouse model in vivo. (A) Combined treatment with PIK-93 and anti-PDL1 anti- bodies induced T cell activity. CHO- K1 cells were pretreated with 0.1 μM PIK-93, 0.5 μM anti–PD-L1 anti- bodies, or combined treatment for 24 hours and subsequently cocul- tured with Jurkat T cells for another 6 hours. The relative luciferase signal indicates the activity of Jurkat T cells. (B and C) The PD-L1 level in the CT26 mouse cancer cell line is reduced after treatment with PIK-93. Cells were harvested, the cytosolic expression of PD-L1 was measured by immunoblotting (B), and the mPD-L1 level was mea- sured by flow cytometry (C). (D and E) The combined treatment inhibits tumor growth in CT26 syngeneic BALB/c mice (n = 8 for each group) (D) and H1975 xenograft in hu- manized mice (NOD.Cg-Prkdcscid

Article Snippet: On the same day, the mice were injected intraperitoneally with DMSO control + human IgG1 antibody (HG1K, Sino Biological), DMSO control + anti–PD-L1 antibody (10mg/kg; atezolizumab, MedChemExpress), PIK-93 (60 mg/kg) + human IgG1 antibody, or the combination of PIK-93 (60 mg/kg) + anti–PD-L1 antibody (10 mg/kg) once every other day for 2 weeks.

Techniques: Activity Assay, In Vivo, Luciferase, Expressing, Western Blot, Flow Cytometry

Journal: Science advances

Article Title: Small-molecule PIK-93 modulates the tumor microenvironment to improve immune checkpoint blockade response.

doi: 10.1126/sciadv.ade9944

Figure Lengend Snippet: Fig. 7. The combination of PIK-93 with anti–PD-L1 antibodies improves the antitumor response in the CT26 animal model resistant to anti–PD-L1 antibodies. (A) Experimental design of the CT26 animal model resistant to anti–PD-L1 antibodies and timeline of each therapy. (B) The percentages of tumor volume increased show that combination treatment with PIK-93 and anti-PDL1 antibodies significantly inhibited tumor growth. Tumor volume increased (%) = [(V −V0)/V0] * 100. V0 is the volume at the start of PIK-93 treatment. The percentages of tumor volume increased are shown as the means ± SD. The linear mixed model was used to examine whether changes in tumor volume over time were different between DMSO control + anti-mouse PD-L1 antibody (6 mg/kg) and the combination of PIK-93 (60 mg/kg) + anti-mouse PD-L1 antibody (6 mg/kg) groups (n = 4 for each group). Increased tumor volume was significantly different between two groups (***P < 0.001). (C) The combination treatment reduced tumor weight at the time of sacrifice (P = 0.0286). The results in (C) are expressed as means ± SEM. Significant differences between indicated groups were calculated by Student’s t test (*P < 0.05).

Article Snippet: On the same day, the mice were injected intraperitoneally with DMSO control + human IgG1 antibody (HG1K, Sino Biological), DMSO control + anti–PD-L1 antibody (10mg/kg; atezolizumab, MedChemExpress), PIK-93 (60 mg/kg) + human IgG1 antibody, or the combination of PIK-93 (60 mg/kg) + anti–PD-L1 antibody (10 mg/kg) once every other day for 2 weeks.

Techniques: Animal Model, Control

Journal: Science advances

Article Title: Small-molecule PIK-93 modulates the tumor microenvironment to improve immune checkpoint blockade response.

doi: 10.1126/sciadv.ade9944

Figure Lengend Snippet: Fig. 8. The model showing that PIK-93 combined with anti–PD-L1 antibodies blocks the PD-1/PD-L1 pathway in the TME. While PD-L1 on tumor cells suppresses T cell activation by binding to PD-1 on T cells, PD-L1 on tumor-associated macrophages (TAMs) contributes to the overall immunosuppressive microenvironment. PIK-93 treatment decreases PD-L1 proteins via E3 ligase CUL4A-facilitated proteasomal degradation. At the same time, a PIK-93–mediated PD-L1 protein reduction on tumor cells leads to increased T cell activity via PD-1/PD-L1 blockade and IFN-γ–induced PD-L1 abundance was also blocked. On the other hand, the reduction of PD-L1 on M1 macrophages by PIK-93 treatment enhances the cytotoxic activity of M1 macrophages. Last, combined treatment with PIK-93 and anti–PD-L1 antibodies increases T cell activity and immune cell recruitment to inhibit lung cancer growth. JAK-STAT-IRF1, Janus kinase–signal transducer and activator of transcription–interferon regulatory factor 1; IFNGR, interferon-gamma receptor; DDB1, DNA damage-binding protein 1; RBX1, ring-box 1.

Article Snippet: On the same day, the mice were injected intraperitoneally with DMSO control + human IgG1 antibody (HG1K, Sino Biological), DMSO control + anti–PD-L1 antibody (10mg/kg; atezolizumab, MedChemExpress), PIK-93 (60 mg/kg) + human IgG1 antibody, or the combination of PIK-93 (60 mg/kg) + anti–PD-L1 antibody (10 mg/kg) once every other day for 2 weeks.

Techniques: Activation Assay, Binding Assay, Activity Assay