Journal: Neoplasia (New York, N.Y.)

Article Title: Doxil Synergizes with Cancer Immunotherapies to Enhance Antitumor Responses in Syngeneic Mouse Models

doi: 10.1016/j.neo.2015.08.004

Figure Lengend Snippet: Synergistic antitumor responses of Doxil in combination with multiple immunotherapies in an established CT26 tumor model. Balb/C mice bearing established (~ 200-300 mm 3 ) CT26 tumors were randomized by tumor volume and treated with maximally efficacious doses of Doxil (5 mg/kg, days 11 and 19), OX40L FP (2.5 mg/kg; days 14 and 19), α-PD-1 (20 mg/kg; days 11, 14, 19, and 22), α-PD-L1, (30 mg/kg; days 11, 14, 19, and 22), α-CTLA-4 (20 mg/kg; days 14, 19, 22, and 26), and GITRL FP (5 mg/kg; days 14, 19, 22, 26, 29, and 32). The groups were as follows: (A) untreated, (B) Doxil, (C) OX40FP, (D) α-PD-1, (E) α-PD-L1, (F) α-CTLA-4, (G) GITRL FP, (H) Doxil + OX40L FP, (I) Doxil + α-PD-1, (J) Doxil + α-PD-L1, (K) Doxil + α-CTLA-4, and (L) Doxil + GITRL FP. The CR number indicates the number of mice that achieved CR out of 12. # P = .056; * P < .008, Bliss independence test.

Article Snippet: Mouse OX40 ligand fusion protein (OX40L FP), mouse GITR ligand fusion protein (GITRL FP), and rat IgG2a isotype control antibodies were produced by MedImmune.

Techniques:

Journal: Neoplasia (New York, N.Y.)

Article Title: Doxil Synergizes with Cancer Immunotherapies to Enhance Antitumor Responses in Syngeneic Mouse Models

doi: 10.1016/j.neo.2015.08.004

Figure Lengend Snippet: Synergistic antitumor responses of Doxil in combination with PD-1, PD-L1, and CTLA-4 antibodies in the MCA205 syngeneic model. C57BL/6 mice bearing established (~ 100-150 mm 3 ) MCA205 tumors were randomized by tumor volume and treated with maximally efficacious doses of Doxil (5 mg/kg; days 10, 17, and 24), OX40L FP (20 mg/kg; days 10 and 14), α-PD-1 (10 mg/kg; days 10, 14, 17, and 21), α-PD-L1 (20 mg/kg; days 10, 14, 17, and 21), α-CTLA-4 (10 mg/kg; days 10, 14, 17, and 21), and GITRL FP (5 mg/kg; days 10, 14, 17, 21, 24, and 28). The groups were as follows: (A) untreated, (B) Doxil, (C) OX40L FP, (D) α-PD-1, (E) α-PD-L1, (F) α-CTLA-4, (G) GITRL FP, (H) Doxil + OX40L FP, (I) Doxil + α-PD-1, (J) Doxil + α-PD-L1, (K) Doxil + α-CTLA-4, and (L) Doxil + GITRL FP. The CR number indicates the number of mice that achieved CR out of 12. * P < .008, Bliss independence test.

Article Snippet: Mouse OX40 ligand fusion protein (OX40L FP), mouse GITR ligand fusion protein (GITRL FP), and rat IgG2a isotype control antibodies were produced by MedImmune.

Techniques:

Journal: Molecular Cancer Therapeutics

Article Title: Recombinant Newcastle Disease Virus Immunotherapy Drives Oncolytic Effects and Durable Systemic Antitumor Immunity

doi: 10.1158/1535-7163.MCT-20-0902

Figure Lengend Snippet: Combination of NDV with immune checkpoint blockade or T-cell agonists in preclinical models. A, Representation of study design combining local NDVmuGM-CSF administration with antibodies to PD-1 or PD-L1 in the CT26 model. B, Spider plots showing tumor growth kinetics in individual animals in each group ( n = 10 per group), data are representative of three separate experiments. C and D, Kaplan–Meier survival analyses of NDVmuGM-CSF plus C, anti–PD-1 or D, PD-L1 antibody combination therapy compared with monotherapies. E, Schematic representation of study design combining NDVmuGM-CSF administration with mGITRL-FP agonist in the CT26 model. F, Spider plots showing tumor growth kinetics of the individual animals in each group ( n = 10 per group except NDVmuGM-CSF, n = 18). G, Kaplan–Meier survival analysis of NDVmuGM-CSF plus mGITRL-FP agonist combination therapy compared with vehicle control and NDVmuGM-CSF and mGITRL-FP monotherapies. H, Schematic representation of study design combining local administration of NDVmuGM-CSF with mOX40L-FP agonist in CT26 model. I, Spider plots showing tumor growth kinetics of the individual animals in each group ( n = 10 per group except NDVmuGM-CSF and NDVmuGM-CSF + OX40L-FP, n = 18). J, Kaplan–Meier survival analysis of NDVmuGM-CSF plus mOX40L-FP agonist combination therapy compared with monotherapy. CR, complete response; PD, progressive disease; PR, partial response. Significance in survival was assessed by log–rank (Mantel–Cox) test. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Article Snippet: Antibodies to PD-1 (clone RMP4–14; Bio X Cell), PD-L1 (mouse IgG1; AstraZeneca), GITRL-FP (mouse IgG2a; AstraZeneca), and OX40L-FP (mouse IgG1; AstraZeneca) were all administered intraperitoneally.

Techniques: Control

Journal: Molecular Cancer Therapeutics

Article Title: Recombinant Newcastle Disease Virus Immunotherapy Drives Oncolytic Effects and Durable Systemic Antitumor Immunity

doi: 10.1158/1535-7163.MCT-20-0902

Figure Lengend Snippet: NDV infection remodeling of TME leading to immune-mediated antitumor activity. A, Tumor volumes after i.t. dosing with NDVmuGM-CSF in CT26 ( n = 12), B, B16F10 ( n = 10), and C, 4T1 ( n = 10) syngeneic tumor models. Dashed vertical lines represent days of NDVmuGM-CSF injections (5 × 10 8 pfu). CT26 tumor growth data (mean ± SEM) shown as spider plots demonstrating heterogeneous antitumor response. D, Representative immunophenotyping of tumor immune infiltrate from three syngeneic models at 24 hours after third i.t. dose of 5 × 10 8 pfu at 8, 11, and 14 days postimplantation; n = 6 vehicle and 6 treated animals for each model. Data are mean ± SEM log 2 values relative to untreated cell counts. NK = natural killer cells; CD19 = B cells; Mθ = macrophage. E, Representative pseudocolour dot plots showing T reg proportions in treated tumors and presence of NDV vHN protein on infected tumor cell surfaces. F, vHN protein expression (MFI) on cell surface of myeloid cell subsets in TME. Data are mean ± SEM. G, PD-L1 expression (MFI) on the surface of myeloid cell subsets in TME. Data are mean ± SEM. H, Detection of “reinvigorated” (PD-1 + , Eomes + , Ki67 + , Gzmb + ) CD8 + T cells in TME. Data are mean ± SEM; *, P < 0.05. I, Tumor growth plots from control CT26 tumor-bearing animals or rechallenged CT26 “cured” animals after NDVmuGM-CSF treatment with CT26 cells on opposite flank. Data represent two separate experiments (controls, n = 3; rechallenged, n = 6). J, Detection of AH1 reactive T cells from splenocytes isolated from control or previously “cured” NDVmuGM-CSF treated animals following CT26 tumor rechallenge as judged by increased number of IFNγ-positive “spots” following stimulation with (red) or without (blue) AH1 peptide ex vivo . Data are mean ± SEM ( n = 2 per group). K and L, Clonality of TCR sequencing isolated from spleen ( K) and tumor (L). Values approaching 1 indicate a nearly monoclonal population of TCRs (controls, n = 8; virus treated, n = 7). M, Comparison of T-cell clonality from vehicle- and NDVmuGM-CSF–treated tumors. N, Morisita index quantitates the similarity between TCR repertoires in spleen and tumors of vehicle- and NDVmuGM-CSF–treated mice. O, Comparison of shared TCR sequences between individual animals in untreated and treated groups. For all comparisons between two data sets, statistical analysis was performed with unpaired t test with Welch correction. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Antibodies to PD-1 (clone RMP4–14; Bio X Cell), PD-L1 (mouse IgG1; AstraZeneca), GITRL-FP (mouse IgG2a; AstraZeneca), and OX40L-FP (mouse IgG1; AstraZeneca) were all administered intraperitoneally.

Techniques: Infection, Activity Assay, Expressing, Control, Isolation, Ex Vivo, Sequencing, Virus, Comparison

Journal: Molecular Cancer Therapeutics

Article Title: Recombinant Newcastle Disease Virus Immunotherapy Drives Oncolytic Effects and Durable Systemic Antitumor Immunity

doi: 10.1158/1535-7163.MCT-20-0902

Figure Lengend Snippet: Combination of NDV with immune checkpoint blockade or T-cell agonists in preclinical models. A, Representation of study design combining local NDVmuGM-CSF administration with antibodies to PD-1 or PD-L1 in the CT26 model. B, Spider plots showing tumor growth kinetics in individual animals in each group ( n = 10 per group), data are representative of three separate experiments. C and D, Kaplan–Meier survival analyses of NDVmuGM-CSF plus C, anti–PD-1 or D, PD-L1 antibody combination therapy compared with monotherapies. E, Schematic representation of study design combining NDVmuGM-CSF administration with mGITRL-FP agonist in the CT26 model. F, Spider plots showing tumor growth kinetics of the individual animals in each group ( n = 10 per group except NDVmuGM-CSF, n = 18). G, Kaplan–Meier survival analysis of NDVmuGM-CSF plus mGITRL-FP agonist combination therapy compared with vehicle control and NDVmuGM-CSF and mGITRL-FP monotherapies. H, Schematic representation of study design combining local administration of NDVmuGM-CSF with mOX40L-FP agonist in CT26 model. I, Spider plots showing tumor growth kinetics of the individual animals in each group ( n = 10 per group except NDVmuGM-CSF and NDVmuGM-CSF + OX40L-FP, n = 18). J, Kaplan–Meier survival analysis of NDVmuGM-CSF plus mOX40L-FP agonist combination therapy compared with monotherapy. CR, complete response; PD, progressive disease; PR, partial response. Significance in survival was assessed by log–rank (Mantel–Cox) test. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Article Snippet: Antibodies to PD-1 (clone RMP4–14; Bio X Cell), PD-L1 (mouse IgG1; AstraZeneca), GITRL-FP (mouse IgG2a; AstraZeneca), and OX40L-FP (mouse IgG1; AstraZeneca) were all administered intraperitoneally.

Techniques: Control

Journal: Molecular Cancer Therapeutics

Article Title: Recombinant Newcastle Disease Virus Immunotherapy Drives Oncolytic Effects and Durable Systemic Antitumor Immunity

doi: 10.1158/1535-7163.MCT-20-0902

Figure Lengend Snippet: Increased susceptibility to NDV was required to generate potent anenestic antitumor responses, which were entirely dependent on modulation of the host immune system. A, Schematic representation of study design with NDVmuGM-CSF administered in a dual-flank CT26 Rig-I −/− , Ifnar1 −/− tumor model. B, Mean tumor growth in untreated animals (blue lines) and spider plots from individual NDVmuGM-CSF–treated animals (red lines) in both treated tumors (solid lines) and untreated tumors (dashed lines; n = 10 animals per group). CR = complete response; PD = progressive disease; PR = partial response. C, Kaplan–Meier plot of survival proportions in dual flank model. D, Representative image of bioluminescent signal detected in NDV-luciferase–treated animals at 48 hours after a single i.t. administration of virus at 5 × 10 8 pfu ( n = 10). Immunophenotyping of treated and untreated tumors with three i.t. doses of NDVmuGM-CSF was performed on days 8, 11, and 14 after implantation and then analyzed 24 hours later on day 15. E–J, Detection of E, vHN protein and F, PD-L1 from NDVmuGM-CSF on surface of CD45 − tumor cells. F, Detection of PD-L1 on surface of tumor cells. G, Detection of neutrophils (CD11b + , Ly6G hi ) within tumors and H, analysis of vHN levels on the surface of tumor-infiltrating neutrophils. I, Detection of “reinvigorated” T cells and J, T regs within tumors. Blue solid squares, vehicle-treated tumors; blue open squares, untreated tumors; red solid circles, NDVmuGM-CSF–treated tumors; red open circles, untreated tumors. MFI = mean fluorescence intensity. K, Quantification of Ki67 + , CD4 + , and CD8 + T-cell populations in vehicle-treated (solid blue bars) and untreated (shaded blue bars) tumors and in NDVmuGM-CSF–treated (solid red bars) and untreated (shaded red bars) tumors ( n = 8 animals per group). Statistical analyses were performed with an unpaired t test with Welch correction; data are mean ± SEM. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Significance in survival benefit was assessed by log–rank (Mantel–Cox) test. RF, right flank; LF, left flank.

Article Snippet: Antibodies to PD-1 (clone RMP4–14; Bio X Cell), PD-L1 (mouse IgG1; AstraZeneca), GITRL-FP (mouse IgG2a; AstraZeneca), and OX40L-FP (mouse IgG1; AstraZeneca) were all administered intraperitoneally.

Techniques: Luciferase, Virus, Fluorescence