Journal: Science Advances

Article Title: P300/CBP inhibition with inobrodib in combination with gilteritinib and venetoclax targets leukemia stem cells in epigenetic mutant AML

doi: 10.1126/sciadv.aec9305

Figure Lengend Snippet: ( A ) In vivo CCS1477 treatment schematic for survival analyses. CD45.2 + D/F or T/F AML cells were transplanted into sublethally irradiated CD45.1 + WT recipient mice. Treatment was initiated 14 days posttransplant. CCS1477 (20 mg/kg) or vehicle was dosed by mouth (PO) once daily (QD). ( B ) Average ± SEM percent c-Kit + cells in the CD45.2 + peripheral blood (PB) of CCS1477- and vehicle-treated D/F ( n = 7 per treatment) and T/F ( n = 11 per treatment) transplanted AML mice over 2 weeks. Week 0 time point was immediately before the first treatment, followed by weeks 1 and 2 on treatment. Significant differences were evaluated by two-way analysis of variance (ANOVA) Šídák’s multiple comparisons test. Average ± SEM ( C ) spleen weight, and numbers of ( D ) CD45.2 + and ( E ) CD45.2 + LSK (Lin − Sca1 + Kit + ) bone marrow cells of D/F ( n = 8 per treatment) or T/F ( n = 8 per treatment) AML mice after 2 weeks of CCS1477 or vehicle. Significant differences were evaluated by unpaired t test. ( F ) Kaplan-Meier survival analysis of CCS1477-treated and vehicle-treated D/F AML (CCS1477, n = 12; vehicle, n = 14) and T/F AML (CCS1477, n = 9; vehicle, n = 9) transplant recipient mice. Overall survival (OS) of moribund D/F AML mice was 94 and 73 days for CCS1477 and vehicle, respectively. OS of moribund T/F AML mice was 51 and 39 days for CCS1477 and vehicle, respectively. Significance determined by the log-rank (Mantel-Cox) test. ( G ) Average ± SEM spleen weight at mouse moribundity of CCS1477- or vehicle-treated D/F ( n = 3 per treatment) or T/F ( n = 3 per treatment) AML mice. Significant differences were evaluated by unpaired t test. Individual data points represent biological replicates. For all panels, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: For CCS1477 monotherapy in vivo studies, CD45.1 + C57BL6/J mice (Charles River Laboratories), 7 to 8 weeks of age, were irradiated with 500 rads before transplantation with either 10 million CD45.2 + D/F AML cells or 2.5 million CD45.2 + T/F AML cells via retro-orbital injection.

Techniques: In Vivo, Irradiation

Journal: bioRxiv

Article Title: Combination of NY-ESO-1-TCR-T-cells coengineered to secrete SiRPα decoys with anti-tumor antibodies to augment macrophage phagocytosis

doi: 10.1101/2023.06.27.546523

Figure Lengend Snippet: ( A ) Schematic of lentiviral vectors encoding SiRPα monomer under 6xNFAT and of a retroviral vector encoding the A97L-TCR. ( B ) Inducible SiRPα monomer expression by A97L-TCR T cells as detected by eGFP expression upon co-culture with target tumor cells (n=3). (C) A375 tumor control curves following ACT with A97L-TCR T cells coengineered to express inSiRPα or CV1 SiRPα monomers under 6xNFAT (n≥6, data representative of 2 independent studies). (D) Evaluation of phagocytosis of A97L-TCR T cells coengineered to express inSiRPα or CV1 SiRPα monomers by BMDMs (n=6). (E) Evaluation of phagocytosis of A97L-TCR T cells coated with inSiRPα or CV1 SiRPα monomers by human MDMs (n=3). (F) Evaluation of phagocytosis of T cells coated with secreted CV1 SiRPα-null-Fc by human MDMs in vitro (n=7). (G) Schematic of retroviral constructs encoding A4-Fc and A4 decoys. (H) Expression of A4-Fc and A4 decoys in transduced mouse OT-I T cells, detected by eGFP (data representative of n≥3 donors). (I) Flow cytometric detection of A4-Fc and A4 monomer binding on OT-I T cells by staining with anti-Fc mAb and anti-mouse CD47 mAbs (data representative of n=3 donors). (J) Schematic of ACT against subcutaneous B16-OVA tumors and ex vivo analysis. (K) Frequency and number of mouse CD45.1 + cells in harvested tissues 6 days post-ACT (n=6, data representative of 2 independent studies). Statistical analysis by one-way analysis of variance (ANOVA) (B and K), two-way ANOVA (C), or unpaired, two-tailed t test (D-F) with correction for multiple comparisons by post hoc Tukey’s test (B and K) or post hoc Sidak’s test (C). ****P< 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05.

Article Snippet: CD45.1 + C57BL/6 female mice were also bred in-house, while female CD45.2 + C57BL/6 mice aged 6-12 weeks old were purchased from Harlan (Harlan, Netherlands).

Techniques: Retroviral, Plasmid Preparation, Expressing, Co-Culture Assay, Control, In Vitro, Construct, Binding Assay, Staining, Ex Vivo, Two Tailed Test

Journal: Nature Communications

Article Title: Lymphodepleting preconditioning impairs host antitumor immunity induced by adoptive T cell therapy in mouse models

doi: 10.1038/s41467-026-71082-y

Figure Lengend Snippet: C57BL/6 mice bearing B16F10-OTI tumors received i.v. transfer of 1 × 10 6 in vitro-activated OTI CD8 + T cells (ACT). Untreated mice (No therapy) were used as controls. a Experimental scheme. Individual tumor growth ( b ) and Kaplan–Meier ( c ) curves for each group: No therapy (black curve), and ACT (blue curves). d – f Tumor and draining lymph node (dLN) infiltrates were analyzed by flow cytometry three days after ACT. d Left panels: Representative plots displaying the frequencies of host (CD45.1 − ) and transferred (OTI CD45.1 + ) CD8 + T cells in live CD45 + cells and their PD-1 and GzmB expression. Middle panels: PD-1 and GzmB expression in host CD8 + T cells and the frequencies of each subpopulation: PD-1 + GzmB − , PD-1 + GzmB + , PD-1 − GzmB − , and PD-1 − GzmB + . Right panels: PD-1 and GzmB expression in transferred OTI CD8 + T cells. e Quantifications of CD45 + cells (percentage of total live cells), transferred OTI CD8 + T cells, and PD-1 + GzmB - and PD-1 + GzmB + host CD8 + T cells as percentages of live CD45 + cells. f Representative plot displaying PD-1 and GzmB expression in host CD8 + T cells. Quadrants define PD-1 + GzmB - (orange), PD-1 + GzmB + (red), and PD-1 − GzmB − (light green). These subpopulations are further characterized by PD-1 and TCF-1 expression and histograms of TIM3, TOX, CD44, CD62L, CD69, CD103, and CD39. g Left panels: Representative plots displaying the frequencies of host (CD45.1 − ) and transferred (OTI CD45.1 + ) CD8 + T cells in live CD45 + cells. Middle panels: PD-1 and GzmB expression in host CD8 + T cells. Right panels: PD-1 and GzmB expression in transferred OTI CD8 + T cells. h Quantifications of OTI, host CD8 + T cells, and PD-1 + CD8 + T cells as a percentage of CD45 + live cells. c Survival Kaplan–Meier curves from two independent experiments. Statistical significance was assessed using a two-sided log-rank Mantel–Cox test **** P < 0.0001. e , h Pooled data from two independent experiments, No therapy n = 8; ACT n = 8. Bars are the mean ± SEM. Statistical significance was assessed using two-sided unpaired Mann–Whitney test * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. CR complete response.

Article Snippet: C57BL/6/J wild-type (CD45.2), B6.129S7- Rag1 tm1Mom /J (RAG1KO), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OTI), CBy.SJL(B6)- Ptprc a /J (CD45.1), B6.129S2-Cd207 tm3(DTR/GFP)Mal /J (Langerin-DTR), B6.Cg-Thy1a/Cy Tg(TcraTcrb)8Rest/J (PMEL-1), B6.129S-Tnftm1Gkl/J (TNFKO) mice were purchased from Jackson Laboratories.

Techniques: In Vitro, Flow Cytometry, Expressing, MANN-WHITNEY

Journal: Nature Communications

Article Title: Lymphodepleting preconditioning impairs host antitumor immunity induced by adoptive T cell therapy in mouse models

doi: 10.1038/s41467-026-71082-y

Figure Lengend Snippet: C57BL/6 mice bearing B16F10-OTI tumors received i.v. transfer of 1 × 10 6 in vitro activated OTI CD8 + T cells (ACT). Untreated mice (No therapy) were used as controls. A group of mice received three i.p. daily doses of αCD8 antibodies starting one day after tumor challenge. a Experimental timeline. b , c Tumor infiltrates were analyzed by flow cytometry three days after ACT. b Representative dot plots displaying the frequencies of host (square, CD45.1 − ) and transferred (ellipse, OTI CD45.1 + ) CD8 + T cells in live CD45 + cells for each group: No therapy, ACT, and αCD8 + ACT. c Quantification of transferred OTI (left panel) and host (right panel) CD8 + T cells as the percentage of live CD45 + cells for each group: No therapy (black bars), ACT (blue bars), αCD8 + ACT (red bars). Individual tumor growth ( d ) and Kaplan–Meier ( e ) curves for each group: No therapy (black curves), ACT (blue curves), and αCD8 + ACT (red curves). c One experiment, No therapy n = 5; ACT n = 5; αCD8 + ACT n = 6. Bars represent mean ± SEM. Statistical significance was assessed using a two-sided unpaired Mann–Whitney test, ** P < 0.01. e Data from three independent experiments. Statistical significance was assessed using a two-sided log-rank Mantel–Cox test, **** P < 0.0001. CR complete response.

Article Snippet: C57BL/6/J wild-type (CD45.2), B6.129S7- Rag1 tm1Mom /J (RAG1KO), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OTI), CBy.SJL(B6)- Ptprc a /J (CD45.1), B6.129S2-Cd207 tm3(DTR/GFP)Mal /J (Langerin-DTR), B6.Cg-Thy1a/Cy Tg(TcraTcrb)8Rest/J (PMEL-1), B6.129S-Tnftm1Gkl/J (TNFKO) mice were purchased from Jackson Laboratories.

Techniques: In Vitro, Flow Cytometry, MANN-WHITNEY

Journal: Nature Communications

Article Title: Lymphodepleting preconditioning impairs host antitumor immunity induced by adoptive T cell therapy in mouse models

doi: 10.1038/s41467-026-71082-y

Figure Lengend Snippet: C57BL/6 mice bearing B16F10-OTI tumors received 300 mg/kg cyclophosphamide (Cy) i.p. on days 4 and 6 after tumor challenge as a lymphodepleting preconditioning regimen. One day later, mice received i.v. transfer of 1 × 10 6 or 0.5 × 10 6 in vitro activated OTI CD8 + T cells as optimal and suboptimal ACT models, respectively. a Experimental timeline. b , c Flow cytometry analysis of tumor-infiltrating lymphocytes three days after ACT. b Representative dot plots displaying the frequencies of host (CD45.1-) and transferred (OTI CD45.1 + ) CD8 + T cells in live CD45 + cells for each group: No therapy, Cy, ACT, Cy + ACT, Suboptimal ACT, and Cy + Suboptimal ACT. c Absolute (cells/g tumor) and relative (on CD45 + cells) quantifications of transferred OTI and host CD8 + T cells in the tumor for each group shown in ( b ). Individual tumor growth ( d ) and Kaplan–Meier ( e ) curves for each group: No therapy (black curves), ACT (blue curves), and Suboptimal ACT (purple curves), Cy (gray curves), Cy + ACT (red curves), and Cy + Suboptimal ACT (green curves). f , g Mice that rejected tumors were rechallenged with 1 × 10 6 wild-type B16F10 in the opposite flank. Individual tumor growth ( f ) and Kaplan–Meier ( g ) curves of rechallenged mice for each group: ACT (blue curves), Cy + ACT (red curves), and Cy + suboptimal ACT (green curves). c One experiment, No therapy n = 5; Cy n = 5; Cy + ACT n = 5; Suboptimal ACT n = 5; Cy + Suboptimal ACT n = 5. Bars represent mean ± SEM. Statistical significance was assessed using a two-sided unpaired Mann–Whitney test * P < 0.05, ** P < 0.01. Kaplan–Meier curves showing the survival from two independent experiments ( e ), and from one experiment ( g ). Statistical significance was assessed using two-sided log-rank Mantel–Cox test * P ˂ 0.05, ** P ˂ 0.01, *** P < 0.001, and **** P < 0.0001. CR complete response, TP total protection.

Article Snippet: C57BL/6/J wild-type (CD45.2), B6.129S7- Rag1 tm1Mom /J (RAG1KO), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OTI), CBy.SJL(B6)- Ptprc a /J (CD45.1), B6.129S2-Cd207 tm3(DTR/GFP)Mal /J (Langerin-DTR), B6.Cg-Thy1a/Cy Tg(TcraTcrb)8Rest/J (PMEL-1), B6.129S-Tnftm1Gkl/J (TNFKO) mice were purchased from Jackson Laboratories.

Techniques: In Vitro, Flow Cytometry, MANN-WHITNEY