Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schematic illustration of US-triggered deep-tissue activation of SPINs to release immunomodulators. b Schematic illustration of sonodynamic activation of SPINs to debulk tumor, enhance tumor immunogenicity, and release immunomodulators in situ as well as synergetic action of IDO inhibition and PD-L1 blocking on enhancing antitumor immunity with alleviated irAEs relative to free-drug administration.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Activation Assay, In Situ, Inhibition, Blocking Assay

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Chemical structures of amphiphilic semiconducting polymeric modulators and schematic illustration of their self-assembly and surface modification to form SPINs. b The molar ratios of each component in different SPINs. c Zeta potentials and hydrodynamic sizes of different SPINs in 1× PBS buffer (pH = 7.4) ( n = 4). d Photographs of erythrocytes after incubation with 1× PBS buffer (negative control), 1% Triton X-100 (positive control), and 1× PBS buffer containing SPINs at the concentration of 100 µg/mL for 2 h, followed by centrifugation. e Hemolysis percentages of erythrocytes after incubation with SPINs at different concentrations for 2 h ( n = 4). f Schematic illustration of US irradiation of SPIN D2 solutions covered with a pork tissue. g ESR spectra of 1 O 2 for SPIN D2 (20 µg/mL) after US irradiation (1.2 W/cm 2 , 3 min) without or with coverage of pork tissues at different thicknesses. h Release profiles of aPD-L1 and NLG919 from SPIN D2 (40 µg/mL) after US irradiation for different time ( n = 4). i PD-L1/PD-1 binding activity assay after treatment with free aPD-L1 or SPIN D2 (40 µg/mL) with or without US irradiation ( n = 4). SPIN D2 – US versus SPIN D2 + US: P < 0.0001. Statistical significance was calculated via a two-tailed Student’s t test. *** P < 0.001. In ( g – i ), the power intensity of US irradiation was 1.2 W/cm 2 (1.0 MHz, 50% duty cycle). Data are presented as mean values ± SD. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Modification, Incubation, Negative Control, Positive Control, Concentration Assay, Centrifugation, Irradiation, Binding Assay, Activity Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schedule for the establishment of primary and distant tumors, triple systemic injection of SPINs (0.2 mL, 0.6 mg/mL) via tail vein, US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min), and analysis of immune responses. b , c Relative tumor volumes of primary ( b ) and distant ( c ) tumors of Panc02 tumor-bearing C57BL/6 mice ( n = 6) after systemic injection of saline, free-drug mixture (on day 0, 3, and 6, 4 mg/kg body weight for NLG919 and aPD-L1), or SPIN D2 (0.2 mL, 0.6 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). SPIN D2 + US versus drug + US: P < 0.0001 for primary tumors ( b ); SPIN D2 + US versus drug: P < 0.0001 for distant tumors ( c ). d Survival curves of Panc02 tumor-bearing C57BL/6 mice ( n = 10) receiving different treatments as indicated. e Schematic illustration of treatment of rechallenged tumor mouse models using SPINs. f Growths of rechallenged tumors in Panc02 tumor-bearing mice after injection of saline or SPIN D2 (0.2 mL, 0.6 mg/mL) with US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min) ( n = 5). Saline versus SPIN D2 + US: P < 0.0001. g The survival curves of Panc02 tumor-bearing C57BL/6 mice after different treatments followed by tumor rechallenge ( n = 10). h Flow cytometry analysis of populations of effector memory T cells in the spleen of Panc02 tumor-bearing C57BL/6 mice after different treatments followed by tumor rechallenge ( n = 4). Saline versus SPIN D2 + US: P = 0.0059. i Differentially expressed gene numbers in tumor tissues of mice after different treatments. j Relative expression of Carl , Hmgb1-ps1 , Hmgb1-ps2 , Cd80 , Cd86 , Cd40 , Pdcd1 , Cd3e , Cd8a , Ifng , Gzmb , Cxcl1 , Cxcl2 , Cxcl9 , Cxcl10 , Cxcl11 , Ccl4 , Ccl5 , Il1b , Il2 , Il6 , Il7 , Il15 , Ido1 , and Cd274 in tumors of Panc02 tumor-bearing mice after different treatments (the experiment was repeated independently five times with similar results). k Unsupervised hierarchical clustering of relative gene expression in tumors of Panc02 tumor-bearing C57BL/6 mice after different treatments ( n = 5). Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Injection, Irradiation, Flow Cytometry, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a Schematic of sono-immunotherapy of subcutaneous pancreatic mouse tumors covered with 5-cm tissue. b , c Relative tumor volumes of primary ( b ) and distant ( c ) tumors of Panc02 tumor-bearing C57BL/6 mice ( n = 5) after systemic injection of saline, free-drug mixture (on day 0, 3, and 6, 4 mg/kg body weight for NLG919 and aPD-L1), SPIN 0 or SPIN D2 (0.2 mL, 0.6 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). The primary tumors were covered with 5-cm tissue under US irradiation. SPIN D2 + US versus SPIN 0 + US: P < 0.0001 for primary tumors ( b ); SPIN D2 + US versus SPIN 0 + US: P < 0.0001 for distant tumors ( c ). d Survival curves of Panc02 tumor-bearing C57BL/6 mice ( n = 10) after different treatments for 60 days. e Schematic of US-mediated deep-tissue sonodynamic therapy of orthotopic pancreatic rabbit tumors. f Radiolabeling stability of 131 I-SPIN 0 after storage in saline or 50% serum at 37 °C for different time ( n = 3). g , h SPECT imaging ( g ) and signal intensity ( h ) of orthotopic pancreatic rabbit tumors after systemic injection of 131 I-SPIN 0 (1.0 mL, 1.5 mg/mL) for different time ( n = 4). The white dotted circle indicated tumors. i Computed tomography (CT) imaging of orthotopic pancreatic rabbit tumors after systemic injection of saline or SPIN 0 (1.0 mL, 1.5 mg/mL) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 30 min). The white dotted circle indicated tumors. j Tumor volume of orthotopic pancreatic rabbit tumors ( n = 3) after treatments as indicated for different days. Saline + US versus SPIN 0 + US: P = 0.0108. k H&E staining images of orthotopic pancreatic rabbit tumors after different treatments. The experiment was repeated independently three times with similar results. l Survival curves of orthotopic pancreatic tumor-bearing rabbits ( n = 4) after different treatments for 20 days. Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; * P < 0.05, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Injection, Irradiation, Radioactivity, Single Photon Emission Computed Tomography, Imaging, Computed Tomography, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Precision cancer sono-immunotherapy using deep-tissue activatable semiconducting polymer immunomodulatory nanoparticles

doi: 10.1038/s41467-022-31551-6

Figure Lengend Snippet: a , b Flow cytometry analysis of percentages of CD3 + CD4 + Th cells ( a ), and CD3 + CD8 + CTLs ( b ) in blood of mice ( n = 4) at 30 day after systemic administrations of saline, SPIN 0 , SPIN D2 (0.2 mL, 1.2 mg/mL) or free-drug mixture (8 mg/kg body weight for NLG919 and aPD-L1) with or without US irradiation (1.0 MHz, 1.2 W/cm 2 , 50% duty cycle, 10 min). Saline – US versus drug − US: P = 0.0023; saline − US versus drug + US: P = 0.0006; drug + US versus SPIN D2 + US: P = 0.0071 for CD3 + CD4 + Th cells ( a ); saline − US versus drug − US: P = 0.0004; saline − US versus drug + US: P = 0.0001; drug + US versus SPIN D2 + US: P = 0.0093 for CD3 + CD8 + CTLs ( b ). c , d Flow cytometry analysis of percentages of CD3 + CD4 + Th cells ( c ), and CD3 + CD8 + CTLs ( d ) in spleen of mice ( n = 4) after different treatments for 30 days. Saline − US versus drug − US: P = 0.0008; saline − US versus drug + US: P = 0.0005; drug + US versus SPIN D2 + US: P = 0.0015 for CD3 + CD4 + Th cells ( c ); saline − US versus drug − US: P = 0.0001; saline − US versus drug + US: P = 0.0002; drug + US versus SPIN D2 + US: P = 0.0049 for CD3 + CD8 + CTLs ( d ). e Representative H&E staining images of liver after 30 days of treatments in different groups (white arrows indicate the infiltrated lymphocytes). The experiments were repeated independently three times with similar results. f Heatmap to show relative fold of cytokine levels in serum of mice after different treatments for 30 days relative to those in saline control group. g , h Serum levels of ALT ( g ) and AST ( h ) in mice ( n = 5) after different treatments for 30 days. Saline − US versus drug − US: P = 0.0010; saline − US versus drug + US: P = 0.0020; drug + US versus SPIN D2 + US: P = 0.0054 for ALT ( g ); saline − US versus drug − US: P = 0.0001; saline − US versus drug + US: P < 0.0001; drug + US versus SPIN D2 + US: P = 0.0013 for AST ( h ). i Summary comparison of the antitumor immunity and irAEs between SPIN D2 -mediated sono-immunotherapy and free-drug treatment. Data are presented as mean values ± SD. Statistical significance was calculated via two-tailed Student’s t test; ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Mouse PD-1[biotinylated]: PD-L1 inhibitor screening assay kit was purchased from BPS Bioscience (San Diego, CA, USA).

Techniques: Flow Cytometry, Irradiation, Staining, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting OAS3 for reversing M2d infiltration and restoring anti-tumor immunity in pancreatic cancer

doi: 10.1007/s00262-024-03898-w

Figure Lengend Snippet: METTL3 plays a pivotal role in preserving the immunosuppressive function of TAMs. A PD-L1 expression in sh-NC Mφ and sh-METTL3 Mφ under CM culture conditions ( n = 3). B ELISA measurements of IL-10 ( B ), VEGF-A ( C ), and IL-6 ( D ) release under different culture conditions in sh-NC Mφ and sh-METTL3 Mφ ( n = 3). C TNF + GZMB + CD8 + T cells in different groups under CM culture conditions ( n = 3). D The effect of TAMs culture supernatants on the angiogenic capacity of HUVECs ( n = 3). E DMSO or STM2457 pretreated BMDMs mixed with PANC-02 cells were inoculated subcutaneously into Balb/c-Nude mice. Images of excised tumors and tumor growth curves were shown ( n = 5). F PANC-02 cells were inoculated subcutaneously with oil, STM2457, or clodronate into pretreated C57BL/6 J mice ( n = 4). G IF showing CD163 + M2 and CD8 + T cell infiltration in tumor tissues from C57BL/6 J mice. *, P < 0.05; **, P < 0.01, ***, P < 0.001, ns., no significance

Article Snippet: These mice were then treated with Mφ or sh-OAS3Mφ, either alone or in combination with Gem (20 mg/kg, 3 times a week) (MCE, HY-B0003) and anti-PD-L1 mAb (100 mg/kg, MCE, HY-155959).

Techniques: Preserving, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting OAS3 for reversing M2d infiltration and restoring anti-tumor immunity in pancreatic cancer

doi: 10.1007/s00262-024-03898-w

Figure Lengend Snippet: Specific OAS3 inhibition in macrophage impaired M2 immunosuppressive function in vivo. A WB images verifying the knockdown effect of OAS3. B Flow cytometry analysis showing the polarization level of CD163 + CD86 − M2 cells in sh-NC and sh-OAS3 Mφ under normal or CM conditions ( n = 3). C IF analysis showing the expression levels of CD163 in Mφ treated with/without CM ( n = 3). D Flow cytometry analysis showing PD-L1 expression levels in sh-NC or sh-OAS3 Mφ under different culture conditions. E ELISA measurements of IL-10 (left) and VEGF-A (right) secretion levels in sh-NC and sh-OAS3 Mφ under different culture conditions ( n = 3). F TNF + GZMB + CD8 + T cells in different groups under CM culture conditions ( n = 3). G The effect of culture supernatants from Mφ or sh-OAS3 Mφ on the angiogenic capacity of HUVECs ( n = 3). *, P < 0.05; **, P < 0.01, ***, P < 0.001

Article Snippet: These mice were then treated with Mφ or sh-OAS3Mφ, either alone or in combination with Gem (20 mg/kg, 3 times a week) (MCE, HY-B0003) and anti-PD-L1 mAb (100 mg/kg, MCE, HY-155959).

Techniques: Inhibition, In Vivo, Knockdown, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Targeting OAS3 for reversing M2d infiltration and restoring anti-tumor immunity in pancreatic cancer

doi: 10.1007/s00262-024-03898-w

Figure Lengend Snippet: Specific OAS3 inhibition in macrophage enhances the efficacy of gemcitabine and anti-PD-L1 mAb treatment in PC in vivo. A IF showing OAS3 and CD8 + T cells in tissues from PC patients. B Intraperitoneal injection of human PBMCs to establish a humanized mouse model of the immune system and construction of a pancreatic cancer CDX model. C In the pancreatic cancer CDX (PANC-1) model, intraperitoneal injection of gemcitabine and PD-L1 mAb or peritumoral injection of macrophages; excised tumor images and tumor growth curves are shown ( n = 5). D Flow cytometry analysis showing the changes in the proportion of CD8 + T cells in peripheral blood of CDX (PANC-1) model mice from different groups ( n = 5). E IF showing CD163 + M2 and CD8 + T cells infiltration in tumor tissues of CDX (PANC-1) model mice. *, P < 0.05; **, P < 0.01, ***, P < 0.001

Article Snippet: These mice were then treated with Mφ or sh-OAS3Mφ, either alone or in combination with Gem (20 mg/kg, 3 times a week) (MCE, HY-B0003) and anti-PD-L1 mAb (100 mg/kg, MCE, HY-155959).

Techniques: Inhibition, In Vivo, Injection, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Antrodia cinnamomea Formula Suppresses Prostate Cancer Progression via Immune Modulation and PD-1/PD-L1 Pathway Inhibition

doi: 10.3390/ijms26062684

Figure Lengend Snippet: Effects of XZF on PD-L1 expression and PD-L1/PD-1 interaction in prostate cancer cells. ( A ) PC-3 cells were treated with ethanol (control), XZF (100 or 200 μg/mL), IFN-γ (10 ng/mL), or IFN-γ combined with XZF (200 μg/mL) for 24 h, followed by total protein extraction. ( B ) Densitometric analysis of PD-L1 normalized to Vinculin in PC-3 cells, expressed as fold change relative to the control. ( C ) DU145 cells were subjected to the same treatment conditions, followed by protein extraction. ( D ) Densitometric analysis of PD-L1 normalized to Vinculin in DU145 cells, expressed as fold change relative to the control. ( E ) PD-L1/PD-1 interaction was evaluated using a homogeneous assay following treatment with ethanol (control), XZF (200 μg/mL), atezolizumab (1 μg/mL), or a combination of atezolizumab and XZF. Data are presented as mean ± S.E.M. from three independent experiments. * p < 0.01 vs. control.

Article Snippet: The PD-1/PD-L1 interaction was assessed using a homogeneous binding assay kit (BPS Bioscience Inc., San Diego, CA, USA; catalog number: 72014) as previously described [ ].

Techniques: Expressing, Control, Protein Extraction