Shattuck Labs, Inc. · NASDAQ:STTK
Shattuck Labs — 3rd Gamma Delta T Therapy Summit 2022
28 slides from Shattuck Labs's conference presentation, Jul 27, 2022
Slide 1: Bispecific T-cell Engager Containing Heterodimeric BTN2A1 and BTN3A1 Promotes Targeted Activation of Vγ9Vδ2+ T cells
Bispecific T-cell Engager Containing Heterodimeric BTN2A1 and BTN3A1 Promotes Targeted Activation of V9V2+ T cells Anne Lai, Ph.D. 3 rd Gamma Delta T Therapy Summit July 27, 2022
Slide 2: Forward-Looking Statements
Forward-Looking Statements This presentation contains “forward-looking statements” within the meaning of the federal securities laws, which statements are subject to substantial risks and uncertainties and are based on our estimates and assumptions. All statements, other than statements of historica
Slide 3: Outline
Outline Overview of the role of butyrophilins (BTNs) in activation of T cells Introduction of Gamma Delta T Cell Engager (GADLENTM) Platform: Leveraging heterodimeric BTNs to harness the cytolytic activity of T cells Proof of Concept: Functional characterization of heme-targeting GADLEN molecule
Slide 4: Shattuck Labs (STTK) Overview
Shattuck Labs (STTK) Overview COMPANY CAPABILITIES PLATFORMS Clinical-stage biotech Experienced R&D, company developing next clinical and technical ARC GADLEN generation bifunctional operations teams (Agonist- (Gamma Delta fusion proteins across offices in Redirected T Cell Engager) Durham, North Ca
Slide 5: Targeting Gamma Delta (γδ) T Cells for Immunotherapy
Targeting Gamma Delta (𝛾𝛿) T Cells for Immunotherapy At the Nexus of Innate and Adaptive Immunity Host Immunity Innate immunity Adaptive immunity (rapid response) (slow response) Key Attributes of 𝛾𝛿 T cells ▪ Shared innate and adaptive features ▪ Superior cytotoxic and serial killing potential…
Slide 6: Role of Butyrophilin Proteins in γδ T cell Function
Role of Butyrophilin Proteins in 𝛾𝛿 T cell Function ▪ The ligands responsible for activating 𝛾𝛿 T cells have long remained a mystery until recently ▪ A conserved role of heterodimeric butyrophilns (BTNs) in modulating function of multiple gdT cell subsets ▪ Heterodimerization of BTN2A1 and…
Slide 7: Basis for the GADLEN Design
Basis for the GADLEN Design Butyrophilins are Natural Ligands that Activate V𝛾9𝛿2 T cells The GADLEN Design: ▪ Activation of V𝛾9𝛿2 T cells; main 𝛾𝛿 population in blood and many tumors ▪ Independent of phosphoantigen levels in cancer cells ▪ Independent of native butyrophilin expression ▪…
Slide 8: GADLEN: Butyrophilin Heterodimer-Based Engager Format
GADLEN: Butyrophilin Heterodimer-Based Engager Format Using Antigen-Targeted scFv Domains to Tether BTN Heterodimers to the Surface of Tumor Cells GADLEN construct includes an active BTN heterodimer BTN2A1 and BTN3A1 BTN2A1+3A1 heterodimer: Heterodimer specifically targets Stimulates 𝛾𝛿 T Cells…
Slide 9: GADLEN: Mechanism of Action
GADLEN: Mechanism of Action GADLEN ➔ BTN heterodimer with costimulation (signal 2) leads to activation of V𝛾9𝛿2 T Cells to promote killing of target cells 9
Slide 10: Characterization of Heme-Targeting GADLEN (Proof-of-Concept)
Characterization of Heme-Targeting GADLEN (Proof-of-Concept) ➔ CD19 and CD20 GADLEN molecules targeting CD19+/CD20+ lymphoma 10
Slide 11: Presence of BTN2A1 and BTN3A1 Heterodimers in GADLEN Construct
Presence of BTN2A1 and BTN3A1 Heterodimers in GADLEN Construct Human BTN2A1/3A1-Fc-CD19scFv BTN2A1/3A1-Fc-CD19scFv BTN3A1-Fc-CD19scFv → Only GADLEN construct containing both BTN2A1 and BTN3A1 heterodimer, but not BTN3A1 homodimer, can be detected using 11 dual-targeting MSD assay
Slide 12: GADLEN Binds to T Cells and Targets Tumor Cells
GADLEN Binds to T Cells and Targets Tumor Cells Human BTN2A1/3A1-Fc-CD19scFv Binding to CD19+ Lymphoma Cells Binding to 𝛾𝛿T Cells 12
Slide 13: GADLEN Specifically Binds the Vγ9 Subunit of T-Cell Receptor
GADLEN Specifically Binds the V9 Subunit of T-Cell Receptor Human BTN2A1/3A1-Fc-CD19scFv V9V2+ T cells 100 Thermo TCR g C C0 0 f cs F C A, C A subset 1 Thermo TCR D C C0 01 f cs F C A, C A subset Thermo no stain C1 C01 00 f cs F C A, C A subset 0 0 % Max. Count D Tcells no stain A01 001 f cs F
Slide 14: GADLEN Selectively Targets Vγ9Vδ2+ T Cells
GADLEN Selectively Targets V9V2+ T Cells Human BTN2A1/3A1-Fc-CD19scFv CD3+V2+ CD3+V1+ CD3+CD8+ % Max. Count GADLEN Binding ➔ Cell binding analysis in PBMC: GADLEN specifically binds to Vd2+ T cells but not Vd1+ or other T cell subsets 14
Slide 15: In Vitro Assays Used to Confirm GADLEN MOA
In Vitro Assays Used to Confirm GADLEN MOA Human BTN2A1/3A1-Fc-CD19scFv Assessed in a tumor cell- free system 𝛾𝛿T cell ▪ CD107a expression by Flow Cytometry Activation ▪ Cytokine production by Flow and ELISA Assessed in a co-culture system Tumor cell ▪ AnnexinV staining by killing Flow Cytometry…
Slide 16: GADLEN Activates T Cells in the Presence of Costimulation
GADLEN Activates T Cells in the Presence of Costimulation Human BTN2A1/3A1-Fc-CD19scFv T Activation (Tumor Cell-Free Assay) γδT Degranulation (CD107a) T Degranulation (CD107a) 2A1/3A1 GADLEN only GADLEN + anti-NKG2D 3A1/3A1 GADLEN + anti-CD28 2A1/2A1 Untreated costim only anti-NKG2D only anti-
Slide 17: GADLEN Activates T Cells in the Presence of Costimulation
GADLEN Activates T Cells in the Presence of Costimulation Human BTN2A1/3A1-Fc-CD19scFv T Activation (Tumor Cell-Free Assay) IFN TNF IFN𝛄 TNF𝛂 2A1/3A1 2A1/3A1 GADLEN only GADLEN GADLEN + anti-NKG2D GADLEN + anti-NKG2D 3A1/3A1 3A1/3A1 GADLEN + anti-CD28 GADLEN + anti-CD28 2A1/2A1 2A1/2A1…
Slide 18: BTN Mediated Signal 1 and CD Mediated Signal 2 Determine Vγ9Vδ2 Activation
BTN Mediated ‘ ignal 1’ and CD Mediated ‘ ignal ’ Determine 9V2 Activation Human BTN2A1/3A1-Fc-CD19scFv Jurkat 76 (TCR-) engineered to express V9V2 TCR Jurkat 76 Activation TCRV9 TCRV2 CD3 TEG activation TEG activation 80 % of Max. Count 80 Untreated anti-NKG2D 60 60 anti-CD28 %CD69+ %CD69+ GA
Slide 19: GADLEN Promotes Targeted Killing of Tumor Cells
0 0 GADLEN Promotes Targeted Killing of Tumor Cells 0 0 Human BTN2A1/3A1-Fc-CD19scFv and BTN2A1/3A1-Fc-CD20scFv 0 0 0 0 0 0 𝛾𝛿T and Raji (CD19+/CD20+ Lymphoma) Co-Culture, 4hrs CD28 Ligands on Raji 10 0 10 10 10 10 0 10 10 (E:T ratio = 1) 100 100 ✱✱ 100 100 0 0 ✱ Isotype 0 0 0 0 % Tumor Cell…
Slide 20: GADLEN-Activated Vγ9Vδ2 T Cells Secrete Key Pro-Inflammatory Cytokines
GADLEN-Activated V𝛾9𝛿2 T Cells Secrete Key Pro-Inflammatory Cytokines 𝛾𝛿T and Raji (CD19+/CD20+ Lymphoma) Co-Culture, 4hrs (E:T ratio = 1) IFN TNF IFN𝛄 TNF𝛂 anti-CD277 (10 μg/mL) Untreated anti-CD277 (10 μg/mL) Un CD19-GADLEN (100 μg/mL) CD19-GADLEN (100 μg/mL)CD19-GADLEN (1 μg/mL) CD…
Slide 21: Blocking of Costimulatory Ligands on Tumor Cells Partially Attenuated GADLEN-Mediated T Cell Killing
Blocking of Costimulatory Ligands on Tumor Cells Partially Attenuated GADLEN-Mediated T Cell Killing Partial inhibition of tumor killing suggests the role of IFN CD19-GADLEN IFN CD20-GADLEN additional co- ✱✱ ✱✱ stimulatory 60000 80 ✱✱ 60000 100 ✱ ✱✱ ✱ ✱✱ Untreated No Treatment ✱ Untreated Untrea
Slide 22: GADLEN Selectively Targets Tumor Cells Expressing Target Antigen
0 GADLEN Selectively Targets Tumor Cells Expressing Target Antigen 0 Human BTN2A1/3A1-Fc-CD19scFv 0 𝛾𝛿T and K562 Co-Culture, 4hrs 0 NKG2D Ligands on K562 γδT andratio (E:T K562 Co-culture = 1) 10 0 10 10 10 % Tumor Cell Apoptosis 25 ✱✱ 100 Untreated 20 CD19-GADLEN 0 Isotype 15 0 MICA/B ULBP1 10 0…
Slide 23: Expression of Costimulatory Ligands (CD80 and CD86) Enables GADLEN Targeting of Human B Cells
Expression of T Costimulatory Ligands (CD80 and CD86) Enables GADLEN Targeting of Human B Cells T and purified B cell co-culture, 4 hrs E:T = 2:1 CD19-GADLEN CD20-GADLEN ✱✱✱ 60 100 ✱✱✱ No Treatment Untreated % Apoptosis % Apoptosis 80 GADLEN GADLEN 40 60 40 20 20 0 0 B cells only γδT+B cells B c
Slide 24: Selective In Vivo Depletion of Human Target Cells by CD20 GADLEN
Selective In Vivo Depletion of Human Target Cells by CD20 GADLEN E:T Ratio = 1:2 (1 𝛾𝛿 T cell per 2 B cells) Experiment Design • CD20-GADLEN depleted >99% CD19+/20+ B cells in vivo (peripheral blood & spleen) • Normal B cells express CD80 and CD86, which provide necessary costim signal (via CD28)…
Slide 25: Efficient Depletion of Human Target Cells at Physiologic γδ Levels
Efficient Depletion of Human Target Cells at Physiologic 𝛾𝛿 Levels E:T Ratio = 1:46 (approx. 1 𝛾𝛿 T cell per 50 B cells) 𝛾𝛿 T cells (Effector) B cells (Target) Non-B cells Donor 1402_CD3-CD19+ Donor 1402_CD3-CD19- CD3- CD20+ CD3- CD20- 105 106 Total cell count in spleen Total cell count in…
Slide 26: Summary of GADLEN Platform
Summary of GADLEN Platform • Activates 𝛾𝛿 T cells with on-target binding to antigen expressed on tumor cells • Identified costimulatory requirement to promote BTN-mediated activation of 𝛾𝛿 T cells • Selectively targets antigen-expressing tumor cells – without evidence of off-target cell killing…
Slide 27: Acknowledgements
Acknowledgements The GADLEN team Suresh de Silva Derek Franklin Arpita Patel Faraha Brewer Kinsley Evans Mahmud Hussain Louis Gonzalez KyungJin Yoo Keith Wilson George Fromm Taylor Schreiber Jenny Gumperz, Univ of Wisconsin- Madison (Advisor and Collaborator) 27
Slide 28: Thank you
Thank you Investor Relations Investorrelations@ShattuckLabs.com
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