Inmune Bio Inc. · NASDAQ:INMB
Inmune Bio — Technical Presentation (March 2021)
15 slides from Inmune Bio's technical presentation, Mar 29, 2021
Slide 1: Exploring the cellular interactions between NK cells, tumor cells and the TME
Professor Mark W Lowdell CSO 1 “Exploring the cellular interactions between NK cells, tumor cells and the TME”
Slide 2: Tumor-derived inhibitory and activating signals; NK-NK interactions and trogocytosis
Tumor - derived Inhibitory signals Activating signals Tumor - derived Inhibitory signals Activating signals Tumor - derived Inhibitory signals Activating signals Activating signals Activating signals NK - Tumor TROGOCYTOSIS NK - NK interactions % of conjugates NK-NK cells 0 20 40 60 80 0.0 0.5 1.0 1
Slide 3: What determines the outcome of NK-tumor ligation?
• Balance of signals – Tumor “sensitivity” • NK:tumor cell ratio • NK cell subset • Prior exposure? – TiM - NK – CiM - NK • Inhibitory microenvironment What determines the outcome of NK - tumor ligation? CD15 - ve K562 NK resistant rNK cell X rNK lack enough activating signals to overcome tumor inhi
Slide 4: E:T determines outcome
E:T determines outcome Low E:T ratios = High NK activation and degranulation – Low lysis CD107a expression on NK cells K 5 6 2 I N B 1 6 0 20 40 60 80 rNK NK+Target[1:2] NK+Target[2:1] NK+Target[5:1] NK+Target[10:1] % o f e x p r e s s i o n CD69 expression on NK cells K 5 6 2 I N B 1 6 0 20 40 60 8
Slide 5: Prior exposure to tumor cells may impair or enhance tumor killing
Prior exposure to tumor cells may impair or enhance tumor killing Priming effect % of expression on NK cells TNFa IFNy Perforin GranzymeB 0 5 10 15 50 60 70 80 90 100 rNK NK+K562 [1:2] NK+INB16 [1:2] % of expression
Slide 6: NK-NK activation following trogocytosis
NK - NK activation following trogocytosis C D 6 9 0 20 40 60 rNK +TpNK +IL15[1ng/mL] % r N K r e c e p t o r e x p r e s s i o n +TpNK(1:10)+IL15 rNK priming by TpNK
Slide 7: NK cells are activated by tumor cell membranes but do not degranulate
CD107a expression on NK cells r N K N K + [ K 5 6 2 ] N K + [ K 5 6 2 2 4 h ] N K + [ K 5 6 2 l y s a t e s ] N K + [ I N B 1 6 ] N K + [ I N B 1 6 2 4 h ] N K + [ I N B 1 6 l y s a t e s ] 0 20 40 60 80 % o f e x p r e s s i o n Killing assay for Raji cells r N K + T a r g e t N K + [ K 5 6 2 ] + T
Slide 8: Does tumor exposure influence NK avidity?
Does tumor exposure influence NK avidity? Kushal Kumar Das - Lumicks Acoustic Node Glass 0.1mm Piezo Element Target tumor cells Primed vs Resting NK cells Ultrasound is used to exert forces completely contactless to NK cells in a microfluidic chip
Slide 9: CD107a expression and killing assay
CD107a expression on NK cells rNK NK+K562[1:2]NK+INB16[1:2] 0 20 40 60 80 100 % of expression Killing assay: % of specific lysis (after 4h) NK+K562 [5:1] NK+INB16 [5:1] 0 20 40 60 80 100 % of specific lysis
Slide 10: CAR-NK versus Mock-NK lysis
Data by Amanda van Vliet CAR - NK Mock - NK 90% lysis 30% lysis
Slide 11: CD107a expression, killing assay and conjugates
CD107a expression on NK cells rNK NK+K562[1:2]NK+INB16[1:2] 0 20 40 60 80 100 % of expression Killing assay: % of specific lysis (after 4h) NK+K562 [5:1] NK+INB16 [5:1] 0 20 40 60 80 100 % of specific lysis E:T [1:1] 15min 30min 45min 60min 0 5 10 rNK TpNK LAK % of conjugates
Slide 12: NK function only needs to be adequate
Alive >2yrs % K562 lysis NK function only needs to be “adequate”
Slide 13: Effects of hypoxia may be transient and tumor cell dependent
Effects of hypoxia may be transient and tumor cell dependent 4 6 16 0 20 40 60 80 100 NK NK (hyp) Time (h) S p e c i f i c C y t o l y s i s ( % ) K562 + rNK RAJI +LAK15 4 6 16 24 48 0 20 40 60 80 100 Time (h) S p e c i f i c C y t o l y s i s ( % ) NK+IL15(10ng/ml) NK+IL15(10ng/ml (hyp) 4 6 16 24 4
Slide 14: Effects of inhibitory cells in the TME
Effects of inhibitory cells in the TME Myeloid - derived suppressor cells Treg Suppression of MLR T r e g : T c o n v 1 : 2 T r e g : T c o n v 1 : 4 T r e g : T c o n v 1 : 8 T r e g : T c o n v 1 : 1 6 T r e g : T c o n v 1 : 3 2 0 20 40 60 80 % o f s u p p r e s s i o n S p e c i f c i c y t o l
Slide 15: Conclusions
Conclusions • NK:tumor cell interactions are multicellular and multifactorial • Degranulation is not a direct correlate of lysis and thus not of exhaustion • Tumor cell conjugation may induce activation without lysis and generate a primed state associated with increased TNF - a and IFN - g and incre