Slide 1: Challenges of NK Immunotherapy of Solid Tumours: Potentiation of intra-tumoral NK function
Challenges of NK Immunotherapy of Solid Tumours: Potentiation of intra - tumoral NK function Professor Mark Lowdell PhD, FRCPath, FRSB 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 expre
Slide 3: NK cell infiltration is associated with improved overall survival in solid cancers
Translational Oncology 14 (2021) 100930 Contents lists available at ScienceDirect Translational Oncology ELSEVIER journal homepage: www.elsevier.com/locate/tranon NK cell infiltration is associated with improved overall survival in solid cancers: A systematic review and meta-analysis ~ Seat Sarah Ne
Slide 4: Be led by the clinical experience
Be led by the clinical experience: 1. Prognostic value of TiNK 2. Association of NK function with OS 3. Histology of primary tumors
Slide 5: Two hypotheses behind NK immunotherapy
• Cancer patient NK cells can be enhanced to mediate autologous tumor killing • Rituximab / Herceptin etc • IL15RA • Ex - vivo activated/expanded NK cells • INmuneBio INKmune • Cancer patient NK cells are terminally defective and must be replaced with NK from healthy donors • Nanktwest • Fate • Take
Slide 6: Challenges of solid tumour NK immunotherapy
Challenges of solid tumour NK immunotherapy Resident TiNK cells • Hypoxia • Cellular inhibition • Tumour - evasion • Loss of adhesion molecules • Loss of activating ligands • Loss of triggering ligands • Upregulation of inhibitory ligands • Secretion of suppressive “factors” • Starvation Adoptive th
Slide 7: The Hypothetical TME Post chemo/radio Rx
The Hypothetical TME Post chemo/radio Rx
Slide 8: Why natural killer cells in triple negative breast cancer?
World l OMAR Civica! Oncology Submit a Manuscript: https:/ /www.t6publishing.com World J Clin Oncol 2020 July 24; 11(7): 464-476 DOI: 10.5306 /wjco.v11.i7.464 ISSN 2218-4333 (online) MINIREVIEWS Why natural killer cells in triple negative breast cancer? Mustafa Abdel-Latif, Rana Ahmed Youness Emilie
Slide 9: How can we overcome the TME?
• NK activation with cytokines or tumor cells • CimlNK or TimlNK ex - vivo • IL2, IL2RA, IL15RA, INKmune in vivo • Enhance immune synapse – increase avidity • Increase lytic function – • Overcome inhibition – activate lytic pathways • Upregulate triggering ligands • Increase secretion of pro - lytic
Slide 10: Peripheral blood NK are impaired by hypoxia whereas TINK are impaired by normoxia
6 0 10 20 30 40 50 60 70 80 90 100 Specific Cytolysis (%) ** * ** * 6 0 10 20 30 40 50 60 70 80 90 100 Specific Cytolysis (%) * Peripheral blood NK are impaired by hypoxia (1% O2) whereas TINK are impaired by normoxia K562 lysis
Slide 11: Cytokine priming enhances TINK cytotoxicity of NK sensitive and resistant tumors
6 16 24 48 0 10 20 30 40 50 60 70 80 90 100 OVCAR3 h+n OCP Time (h) Specific Cytolysis (%) OCP - Ascites rNK OCP - Ascites+IL - 15 OCP - Ascites rNK HYP OCP - Ascites+IL - 15 HYP Cytokine priming (rhIL - 15 10ng/ml) enhances TINK cytotoxicity of NK sensitive & NK resistant tumors OCP - derived rNK c
Slide 12: 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 Effect of NK priming on hypoxia - induced impairment of NK lysis % change from normoxia IL15 treated INKmune treated OCP - Ascites derived NK cells Cytokine priming (rhIL - 15 10ng/ml) enhances TINK cytotoxicity of NK - sensitive OVCAR - 3 Tumor - priming of T
Slide 13: Effects of inhibitory cells in the TME
Effects of inhibitory cells in the TME Myeloid - derived suppressor cells Specifci cytolysis 80 60 40 20 0 100 Treg Suppression of NK - mediated lysis K562 lysis 5:1:1 0 10 20 30 40 % CD107a+ NIL Allo 1 Allo 2 Auto K562 lysis 5:1:1 0 20 40 60 80 100 rNK rNK:CD33 (IL - 6) Allo 1 rNK:CD33 (VEGF) Allo
Slide 14: NK activation is the outcome of enough of the right signals
NK activation is the outcome of “ enough of the right signals” • Ligands in GREEN boxes are activating • The rest are inhibitory • MICA/ULBP, ICAM - 1 and CD15 are enough to prime an NK cell if delivered at the correct density • Blocking all three reduces INKMune priming but doesn’t block completely
Slide 15: Formation of a strong NK:tumor cell synapse is essential for lysis
1. NK recognizes tumor cell 3. NK forms stable immune synapse with tumor cell and NK ligands send activating signals to the NK cell 2. NK makes strong bond to tumor cell 4. NK cell kills tumor cell FORMATION OF A STRONG NK : TUMOR CELL SYNAPSE IS ESSENTIAL FOR LYSIS 1. NK recognizes RESISTANT tumor
Slide 16: Tumour-induced memory can enhance formation of synapse
TUMOUR - INDUCED MEMORY CAN ENHANCE FORMATION OF SYNAPSE 1. In the bloodstream INKmune binds to NK cells 2. The INKmune cell provides activating signals missing from NK - resistant cancer cells 3. The INKmune primed NK cell now has enough signals to form a STRONG bond to the tumor cell 4. INKmune pr
Slide 17: NK potentiation by tumor-priming
NK potentiation by tumor - priming Resting NK don’t bind/kill resistant tumor cells Priming with IL - 15 promotes NK killing INKmune - primed NK form intensely strong bonds to tumor cells SKOV3+CpNK+IL15 SKOV3+CpNK+IL12 SKOV3+CpNK+IL2 SKOV3+rNK SKOV3+CpNK
Slide 18: Tumor-priming increases avidity of NK cell binding to tumor cells
Tumor - priming increases avidity of NK cell binding to tumor cells 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
Slide 19: CD107a expression and killing assay
CD107a expression on NK cells 100 80 60 40 20 0 % of expression Killing assay: % of specific lysis (after 4h) 0 20 40 60 80 100 % of specific lysis E:T [1:1] 0 5 10 rNK TpNK LAK % of conjugates
Slide 20: Effect of whole tumour priming on phenotype
60 40 20 0 80 100 % expression Activation markers rNK NK+INB16 NK+NALM16 80 60 40 20 0 100 % expression Chemokine receptors Maturation markers Inhibitory markers Immune checkpoints Effect of whole tumour priming on phenotype
Slide 21: NK proteomics after INKmune tumor-priming vs IL-15
INB16 upregulates 1461 proteins distinct from IL15 – including Granulysin, ROMO1 & DEFA3 Target cell lysis proteins INB16 upregulates the same proteins as IL15 Mitochondrial survival proteins INB16 upregulates significantly more than IL15 NK proteomics after INKmune tumor - priming vs IL - 15 INB16
Slide 22: MoA of tumor-priming = serial activation of rNK
140 120 100 80 60 40 20 0 Number of contacts (absolute count) 0' 15' 30' 45' 60' 0 10 20 30 40 50 60 Time (minutes) % NK:NK cell conjugates CD16 0 20 40 60 % NK cell expression rNK: rNK rNK:TIML - NK/INB16 A B C D rNK: rNK rNK: TIML - NK/INB16 MoA of tumor - priming = serial activation of rNK 0 2 0
Slide 23: NK function only needs to be adequate
Alive >2yrs % K562 lysis NK function only needs to be “adequate”
Slide 24: INKmune First in Human Trial – MDS/AML
INKmune First in Human Trial – MDS/AML Day 119 data - First Patient treated with INKmune IV INKmune activates tumor killing NK cells and is associated with reduction in BM blast count Lowest dose cohort INKmune treatment at 119 days ▪ NO side effects – NO evidence of alloresponse to INKmune ▪ Eviden
Slide 25: Systemic cytokine response to INKmune
Day 119 data – Systemic cytokine response to INKmune 200 150 100 50 0 250 Concentration [pg/ml] IL - 6 MIP - 1beta MIP - 1alpha Eotaxin TNF 150 100 50 0 Concentration [pg/ml] IL - 12 IL - 15 IL - 2R IL - 8 Screen Day 1 pre D8 D15 D29 D43 D73 D119 80 60 40 20 0 % Activated (CD69+) of total NK
Slide 26: Conclusions
Conclusions • TINK are associated with improved prognosis in multiple cancers • Enhancing TINK function with cytokines ( ex - vivo/in vivo ) or generation of TiML ( ex - vivo/in vivo ) can overcome suppressive TME • Classical regulatory factors in the TME can inhibit NK function in vitro but the eff
Slide 27: Acknowledgements
Acknowledgements • INMB/UCL team • Aga Zubiak • May Sabry • Xenia Charalambous • Helena Arellano Ballestero • Fahim Ghourbandi • Shaun Haran • Andrea Knight • Lumicks • Kushal Kumal Das • Andrea Candelli • INmune Bio Inc – Josh Schoonover – Cory Ellspermann – David Moss – RJ Tesi