Anti-TNF in Melanoma Immunotherapy: From Basic Findings to the Clinic PDF

ANTI-TNF IN MELANOMA IMMUNOTHERAPY: FROM BASIC FINDINGS TO THE CLINIC Bruno Ségui Professor of Cell Biology, School of Pharmaceutical Sciences, Toulouse III-P. Sabatier University Deputy Director of the CARe graduate school (https://care-graduateschool.fr) Co-PI of Melasphinx Team, Cancer...

ANTI-TNF IN MELANOMA IMMUNOTHERAPY: FROM BASIC FINDINGS TO THE CLINIC Bruno Ségui Professor of Cell Biology, School of Pharmaceutical Sciences, Toulouse III-P. Sabatier University Deputy Director of the CARe graduate school (https://care-graduateschool.fr) Co-PI of Melasphinx Team, Cancer Research Center of Toulouse 1037 5071  DISCLOSURE STATEMENT Since 2017: Investigator/speaker/ consultant for Principal investigator MELANFa : A preclinical research program on immunotherapy in mice Exploratory part TICIMEL: A phase 1b clinical trial in advanced melanoma patients MELANFa: A pilot study in advanced melanoma patients  TNF plays a dual role in cancer… including in melanoma Tumor Necrosis Factor Cancer cell death Carswell et al., PNAS 1975 TNF Isolated limb perfusion of recombinant TNF Tumor Microenvironment Endothelial cell death Meyrick et al., Am J Pathol., 1991 Tumor Progression and metastases  TNF plays a dual role in cancer… including in melanoma Tumor Necrosis Factor Tumor-Promoting Factor Balkwill et al., Nat. Rev Cancer 2009 Cancer cell death Cancer cell Carswell et al., PNAS 1975 TNF ü Proliferation ü Migration ü Differentiation Isolated limb ü Resistance perfusion of recombinant TNF Tumor Microenvironment Chronic inflammation ü Angiogenesis Endothelial cell death ü Stromagenesis Meyrick et al., Am J Pathol., 1991 ü Leukocyte infiltration/activity Tumor Progression and metastases  TNF has pleiotropic functions in anti-cancer immune response… including in melanoma Dendritic cell maturation Co-stimulation Kim et al., JI 2001; Ding et al., JI 2011 Adhesion molecules Briscoe et al., JI 1992 TNF Lymphocyte- mediated cytotoxicity Ratner et al. JI. 1993 Anti-cancer immune response  TNF has pleiotropic functions in anti-cancer immune response… including in melanoma Dendritic cell maturation Co-stimulation AICD in CD8 T Kim et al., JI 2001; Ding et al., JI 2011 cells Zheng et al., Nature 1995 way of limiting the CD8 cells Immunosuppression ü MDSC Zhao et al. JCI 2012 Adhesion molecules ü Treg Chen et al., JI 2007 Briscoe et al., JI 1992 ü Breg Schioppa et al. PNAS 2011 TNF Melanoma dedifferentiation (loss of melanocytic antigens) Landsberg et al. Nature 2012 Lymphocyte- mediated cytotoxicity PD-L1 expression Ratner et al., JI 1993 Lin et al. Cancer cell 2016 CD73 expression Reinhardt et al., Cancer Res 2017 Anti-cancer immune response  MELANOMA IMMUNOTHERAPY ipilimumab + nivolumab nivolumab anti PDL-1 Ab ipilimumab anti CTLA 4 Ab ipilimumab + nivolumab combined immune checkpoint inhibitor are better ü 22% of complete response ü 42% of non-responders ü 36% of partial response ü 22% of relapse within 5 years post-induction ü 12% of stable disease ü 95% of immune-related adverse events / 59% of grade 3-4 ü 20% of colitis / corticotherapy ± anti-TNF (infliximab) adverse effects What is the impact of anti-TNF in anti-melanoma immune response and melanoma progression?  2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Investigating the impact of TNF blockade in experimental mouse melanoma model Orthotopic (i.e., intradermal) injection of melanoma cell lines in C57BL/6 WT mice Impact of TNF blockade on: (i) melanoma growth (ii) anti-melanoma immune response (iii) anti-PD-1 therapy  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 202 From the CRCT To the IUCT Back to the CRCT melanoma cell lines- express high major WT mice + B16K1 histocompatibility complex-way of facilitating the CD8 immune response 3) 3) 1200 Excipient ** (mm volume(mm 1000 Etanercept block TNF 800 Tumor volume 600 ** 400 * Tumor 200 0 18 21 24 27 30 Time (days) Etanercept inhibits B16K1 melanoma growth in WT mice and increases CD8+ TIL content  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT WT mice + B16K1 Day 12: % CD8+ TILs by flow cytometry 3) 3) 1200 Excipient ** tumour infiltrating lymphocytes (mm volume(mm 1000 Etanercept TNF-2 R blocker 800 Tumor volume % of total 600 ** 400 * Tumor 200 0 18 21 24 27 30 Vehicule Etanercept Time (days) Etanercept inhibits B16K1 melanoma growth in WT mice and increases CD8+ TIL content  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT WT mice + B16K1 3) 3) 1200 Excipient ** Excipient Etanercept (mm volume(mm 1000 Etanercept 800 Tumor volume 600 ** 400 * Tumor 200 0 18 21 24 27 30 CD8 CD31 MECA79 Time (days) Etanercept inhibits B16K1 melanoma growth in WT mice and increases CD8+ TIL content  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT WT mice + B16K1 CD8 KO mice + B16K1 700 3) 3) 1200 Excipient ** Excipient (mm 600 volume(mm 1000 Etanercept 500 Etanercept 800 Tumor volume 400 600 ** 300 400 * Tumor 200 200 100 0 0 18 21 24 27 30 7 9 11 13 15 17 19 21 23 Time (days) Time (days) TNF facilitates melanoma growth by impairing CD8 T cell-dependent immune response  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Anti-TNF enhances anti-PD-1 therapy against B16K1 melanoma in WT mice double standard treatment of melanoma a Ctrl aTNF Tumor volume (mm3) W T c o n tr o l a n ti-T N F c o n tr o l 1,000 1000 1,000 1000 0/24 0/24 T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) 500 500 500 500 00 00 00 202 0 40 40 60 60 0 0 20 20 40 40 60 60 T im e (d a y s ) T im e (d a y s ) anti TNF aPD-1 aTNF N Fa complete responses W T + a n ti-P D 1 a n ti-T+ + PD-1 a n ti-P D 1 Tumor volume (mm3) 1,000 1000 1,000 1000 7/24 18/24 T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) 500 500 500 500 00 00 00 20 20 40 40 60 60 00 20 20 40 40 60 60 T im e (d a y s ) T im e (d a y s ) Time (Days) Time (Days) Anti-PD-1 +/- anti-TNF: 10 mg/Kg at days 6, 10 and 13  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Anti-TNF enhances anti-PD-1 therapy against B16K1 melanoma in WT mice a b Ctrl aTNF Tumor volume (mm3) Overall survival (%) W T c o n tr o l a n ti-T N F c o n tr o l 100 100 1,000 1000 1,000 1000 P e rc e n t s u rv iv a l 0/24 0/24 Ctrl T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) aPD-1 500 500 505 0 aTNF 500 500 aTNF + aPD-1 00 00 00 202 0 40 40 60 60 0 0 20 20 40 40 60 60 00 T im e (d a y s ) T im e (d a y s ) 0 0 20 20 40 40 60 60 120 80 T im e Time (Days) aPD-1 W T + a n ti-P D 1 aTNF N Fa a n ti-T+ + PD-1 a n ti-P D 1 Tumor volume (mm3) 1,000 1000 1,000 1000 7/24 18/24 T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) 500 500 500 500 00 00 00 20 20 40 40 60 60 00 20 20 40 40 60 60 T im e (d a y s ) T im e (d a y s ) Time (Days) Time (Days) Anti-PD-1 +/- anti-TNF: 10 mg/Kg at days 6, 10 and 13  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Anti-TNF enhances anti-PD-1 therapy against B16K1 melanoma in WT mice a b Ctrl aTNF Tumor volume (mm3) Overall survival (%) W T c o n tr o l a n ti-T N F c o n tr o l 100 100 1,000 1000 1,000 1000 P e rc e n t s u rv iv a l 0/24 0/24 Ctrl T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) aPD-1 500 500 505 0 aTNF 500 500 aTNF + aPD-1 00 00 00 202 0 40 40 60 60 0 0 20 20 40 40 60 60 00 T im e (d a y s ) T im e (d a y s ) 0 0 20 20 40 40 60 60 120 80 T im e Time (Days) aPD-1 W T + a n ti-P D 1 aTNF N Fa a n ti-T+ + PD-1 a n ti-P D 1 Tumor volume (mm3) 1,000 1000 7/24 1,000 1000 18/24 c D10: CD8+ TILs tumour infiltrating lymphocytes T u m o r v o lu m e (m m 3 ) T u m o r v o lu m e (m m 3 ) cells) 500 500 * cells) 500 500 15 ** total of total 10 00 00 (% of 00 20T im e (d a y s40 20 40 60 60 0 0 20 20 40 40 60 60 ) T im e (d a y s ) TILs(% 5 Time (Days) Time (Days) TILs CD8+ CD8+ 0 Anti-PD-1 +/- anti-TNF: 10 mg/Kg αPD-1 - + - + better when we combine αTNF - - + + at days 6, 10 and 13  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Anti-TNF prevents anti-PD-1-induced TIM-3 up-regulation on TILs a CD8+ TILs b ** TIM3 on CD8+ TILs (MFI) 10000 ** 8000 Vehicle 6000 αPD-1 4000 % of max 2000 αTNF 0 αPD-1 - + - + αTNF + αPD-1 αTNF - - + + no significant TIM 3 expression TIM3 immune checkpoint molecule that is upregulated during the synergy of two drugs one of the possible mechanism  I- TNF: AN IMMUNOSUPPRESSIVE CYTOKINE IN MOUSE MELANOMA 2011 2017 2020 2024 From the CRCT To the IUCT Back to the CRCT Anti-TNF prevents anti-PD-1-induced TIM-3 up-regulation on TILs a CD8+ TILs b ** TIM3 on CD8+ TILs (MFI) 10000 ** 8000 Vehicle 6000 αPD-1 4000 % of max 2000 αTNF 0 αPD-1 - + - + αTNF + αPD-1 αTNF - - + + TIM3 c TCGA melanoma d Patients with anti-PD-1 therapy r=0.723 p