Slide 1: Arming NK cells to override immunosuppression in the TME
Arming NK ce l l s to ov e rride immunos u ppre s sio n i n the TME Professor Mark Lowdell CSO INmune Bio Inc.
Slide 2: Forward-looking statements
This presentation contains “forward - looking statements” Forward - looking statements reflect our current view about future events. When used in this presentation, the words “anticipate,” “believe,” “estimate,” “expect,” “future,” “intend,” “plan,” or the negative of these terms and similar express
Slide 3: Tumor-primed human NK cells lyse NK-resistant leukemic targets
% specific lysis ae ; : ; Fi INmuneBi ‘Tumor-Primed Human Natu Killer Cells Lyse NK-Resistant Leukemic Priming of Resting NK Cells Is Killer Ig-like INNATE IMMUNITY Tumor Targets: Evidence of a Two-Stage Process in Resting Receptor Independent but Requires CD15-Mediated CD2 NK Cell Activation’ The J
Slide 4: NK cell specific lysis and expansion data
A B C C tr l D a y 1 C tr l D a y 7 TIML - N K ce ll s / NB1 6 Day 7 TIML - N K ce ll s / NA LM 1 6 Day 7 D E CellTrace TM Violet Fluorescence Cel l C o u n t 0 20 40 60 80 100 % specific lysis of K562 0 20 40 60 80 100 % specific lysis of RAJI 0 2 4 6 NK cell expansion (fold change)
Slide 5: Two-stage priming of allogeneic NK cells in AML: Phase I trial
Two-Stage Priming of Allogeneic Natural Killer Cells for the Treatment of Patients with Acute Myeloid Leukemia: A Phase I Trial Panagiotis D. Kottaridis, Janet North, Maria Tsirogianni, Chloe Marden, Edward R. Samuel, Sam Jide-Banwo, Sarah Grace, Mark W. Lowdell* ‘Table 2. Response after Treatment.
Slide 6: MoA of tumor-priming: serial activation of rNK
140 120 100 80 60 40 20 0 N umbe r o f con t ac t s (absolute count) 0' 15' 30' 45' 60' 0 10 20 30 40 50 60 T im e (m i nutes ) % NK:NK cell conjugates 0 20 40 60 % NK cell expression r N K : r NK rNK:TIM A B C D MoA of tumor - priming = serial activation of rNK 0 2 0 4 0 6 0 r N K r N K + I L 1 5 [
Slide 7: Tumor- and cytokine-primed NK cells show distinct signatures
‘SPLOS | one INmuneBi INNATE IMMUNITY IFN Tumor- and cytokine-primed human natural killer cells exhibit distinct phenotypic and transcriptional signatures ", Agnieszka Zubiak’, Simon P. Hood”, Poppy Simmonds’ , Helena Arellano- ‘Cournoyer', Meghavi Mashar', A. Graham Pockley*, Mark (6) cal sees he I
Slide 8: Effect of whole tumour priming on phenotype
60 40 20 0 80 100 % ex p r e s s i o n Activation markers r N K NK+INB16 NK+NA L M 1 6 8 0 6 0 4 0 2 0 0 100 % expression Ch em oki n e receptors M atu r at i o n markers Inhibitory markers Immune checkpoints Effect of whole tumour priming on phenotype
Slide 9: Effect of NK priming on hypoxia-induced impairment of NK lysis
1 6 2 4 4 8 - 1 0 0 0 1 0 0 2 0 0 3 0 0 OCP - Ascites derived NK cells Effect of NK priming on hypoxia - induced impairment of NK lysis % c h a n g e f r o m no rm o x i a IL15 treated INKmune treated Tumor - priming of TINK enhances killing of NK - resistant SKOV - 3 6 16 24 48 0 10 20 30 40 50 60
Slide 10: Effects of inhibitory cells in the TME
Effects of in h ib i tory cell s i n the TME Myeloid - derived suppressor cells K562 lysis 5:1:1 S p e c i fc i c y t ol ys i s 8 0 6 0 4 0 2 0 0 100 Treg Suppression of NK - mediated lysis 0 1 0 2 0 3 0 4 0 % CD107a+ N I L A l l o 1 A l l o 2 A u t o K562 lysis 5:1:1 0 2 0 4 0 6 0 8 0 100 rNK rNK:C
Slide 11: Gene expression changes after priming
, IL1A, IL1 B , IL3 R A , B , M AFF , M IF, M YC, M B 9 , PS M E2, RG S1 4 , CC L8 , CC L2 0 , CC L F 1 , CSF2, CXC L 1 , CX A M 1, ID O 1, IF NG , IL NF S F 4 , T NF S F 10 0 , CC L 1 , CC L3 , C T N F R SF 9 , T N FS F 1 0 , T R AF D 1 , downre L G A LS 3 , MR C 1 , TY RO B P ( 92 ) TLR 4 Upregula
Slide 12: INKmune is a pseudokine: MOA
I N K M U N E I S A “ P S E U D O K I N E ” MOA - INKmune Œ activates critical genes and pathways for enhanced NK function / survival INB16 upregulates 522 genes distinct from IL2 – including CD70, CXCL10, Stat5 INB16 upregulates 1461 proteins distinct from IL15 – including TIMM29, USP20 Target cell
Slide 13: TpNK proteomics
rNK LAK - 15 CIML - NK INKMUNE - NK TpNK proteomics Proteins significantly increased in TpNK cf CIML - NK
Slide 14: INKmune enhances NK metabolism and survival
I N K M U N E E N H A N C E S N K M E T A B O L I S M & S U R V I V A L Rapid proliferation of NK cells ex - vivo leads to reduced mitochondrial fitness; decreased membrane potential and increased levels of mitochondrial reactive oxygen species (mtROS) Mitochondrial fitness has to be restored for th
Slide 15: INKmune: a multi-dimensional pseudokine
INKmun e : a mult i - dimensional “ pseudokine ” rNK cells (green) added to resistant tumor cells INKmune primed NK cells added to resistant tumor cells Low force causes break of immune synapse and NK collect in clusters Increasing force fails to break immune synapse and NK remain bound to tumor cel
Slide 16: INKmune increases avidity of NK cell binding to tumor cells
16 I N K M U N E I S A “ P S E U D O K I N E ” MO A - I N K m un e Œ I n cr e a s es Avidity of N K Cell Bi ndi n g to Tu m or Cel l s 0.0 5.0 25 . 0 20 . 0 15 . 0 10 . 0 30 . 0 40 . 0 35 . 0 24 . 0 36 . 0 48 . 0 60.0 Time (hr Post - Treatment) 72 . 0 84 . 0 % Cytolysis rNK 20:1 TpNK 5:1 xCelligence
Slide 17: INKmune differentiates resting NK to memory-like NK cells
A M U L T I - D I M E N S I O N A L “ P S E U D O K I N E ” MOA – INKmune Œ Differentiates Resting NK to “Memory - Like” NK Cells Cytokine cocktail (IL - 12/15/18) drives resting NK to memory - like NK ( S ci T ransl M e d 2016 ; Ro m ee ,F enhege r , e t al) 0 5000 10000 15000 NK G 2 C M e F I 0 10
Slide 18: INKmune infusions: patient results
INKmune infusions: D 1 7 th July D 8 14 th July D 15 21 st July All infusions well tolerated, no infusional AEs Episode of neutropenic sepsis ( Stenotrophomonas ) D29 BM 9% blasts RAD1 45% D100 BM 13% blasts RAD1 38% Las t f/u: Reduced transfusion requirements Fully active, regular exercise – ECOG 0
Slide 19: Cytokine concentration data
0 5 0 10 0 15 0 20 0 25 0 0 2 0 4 0 6 0 8 0 Concen t ra ti on [ pg / m l ] Concen t ra ti on [ pg / m l ] M I P - 1 be t a M I P - 1 a l ph a E o t a x i n TNF alpha 0 5 0 10 0 15 0 0 100 0 200 0 300 0 Concen t ra ti on [ pg / m l ] Concen t ra ti on [ pg / m l ] I L - 1 2 I L - 1 5 I L - 2 R IL - 8
Slide 20: Conclusions
C o n c l u s i o n s • NK interaction with some tumor cells provides priming signals which drive a type of ml - NK cell - TpNK • TpNK target hematological and solid tumors and can be generated from patient NK cells • TpNK are not inhibited by self MHC – akin to CIML - NK • TpNK show increased activ
Slide 21: Acknowledgements
Acknowledgements Janet North Dr Ismail Bakhsh Dr May Sabry Dr Xenia Charalambous Aga Zubiak Helena Arellano Ballestero Dr Ben Weil Dr Rita Rego Carla Carvalho Simon Hood Prof Graham Pockley Dr Panagiotis Kottaridis Dr Maria Tsirogianni Dr Kim Orchard Dr Chris Dalley Dr Debbie Richardson Das Kushal D