Innate Pharma Sa Class A · EURONEXT:IPH
Innate Pharma Sa Class A — EORTC CLTF meeting, Torino 2015
65 slides from Innate Pharma Sa Class A's conference presentation, Sep 25, 2015
Slide 1: Neil Smith Memorial Lecture: A novel targeted immunotherapy for CTCL
Neil Smith Memorial Lecture A novel targeted immunotherapy for CTCL Martine Bagot Department of Dermatology and Inserm U976, Hôpital Saint-Louis, Paris, France martine.bagot@sls.aphp.fr EORTC CLTF meeting, Torino, September 25-27, 2015
Slide 2: Slide 2
Slide 3: How can we find new efficient treatments for CTCL patients?
How can we find new efficient treatments for CTCL patients ? • Genomic studies to identify mutations and relevant targets within various signaling pathways • Non specific immunotherapy: unleashing the immune system by releasing its negative regulatory checkpoints • Specific immunotherapy: identifyin
Slide 4: Is it possible to separate reactive T cell clones from tumor T cell clones in CTCL?
Is it possible to separate reactive T cell clones from tumor T cell clones in CTCL skin and blood ?
Slide 5: Is there evidence for Tumor Infiltrating Lymphocytes in CTCL?
Is there evidence for Tumor Infiltrating Lymphocytes in CTCL ?
Slide 6: Isolation of skin Tumor Infiltrating Lymphocytes in CTCL
Isolation of skin Tumor Infiltrating Lymphocytes in CTCL Martine Bagot et al. Blood 1998;91:4331-4341
Slide 7: T cell clones isolated from CTCL may arise from tumor and reactive lymphocytes
T cell clones isolated from CTCL may arise from tumor T lymphocytes but also from reactive T lymphocytes
Slide 8: Establishment of CTCL-derived long-term T cell lines
Establishment of CTCL-derived long-term T cell lines Armand Bensussan Member of International Human Cell Differentiation Molecules Council (CD nomenclature) Test of more than 300 mAbs evaluated during the CDs’ workshops
Slide 9: CD158k/KIR3DL2 is a new phenotypic marker of Sezary cells
6G E blood 2001 97: 1388-1391 —" doi:10.1182/blood.V97.5.1388 CD4* cutaneous T-cell lymphoma cells express the p140-killer cell immunoglobulin-like receptor Martine Bagot, Alessandro Moretta, Simona Sivori, Roberto Biassoni, Claudia Cantoni, Cristina Bottino, Laurence Boumsell and Armand Bensussan C
Slide 10: KIR3DL2 is a phenotypic marker for Sezary cells
KIR3DL2 is a phenotypic marker for Sezary cells KIR3DL2 Patient 1 CD4 CD3 V1 KIR3DL2 CD4 Patient 2 CD3 V8 Clonal leukemic Sézary cells, defined by a single V chain expression, are KIR3DL2+ in patient blood Moins-Teisserenc H, J Invest Dermatol 2014
Slide 11: Leucocyte Receptor Cluster Organization
Leucocyte Receptor Cluster Organization Homme Chr 19 Souris Chr 7
Slide 12: KIR Receptor Family
KIR Receptor Family Adapted from Thielens et al., Curr Opin Immunol. 2012 Apr;24(2):239-45
Slide 13: The KIR3DL2/CD158k receptor
The KIR3DL2/CD158k receptor
Slide 14: Identification of Sezary cells using cytomorphology, flow-cytometry or TCR repertoire
Identification of Sezary cells using either cytomorphology, flow-cytometry or TCR repertoire BERP36 3500" 3000" 4000 6000 8000 10000 2500" 12000 10000 2000" SS MF 1500" CD158k+ T cells/mm3 8000 1000" 500" 6000 0" 12/14/10" 3/11/11" 3/25/11" 7/8/11" 2/2/12" 4/12/12" SCbs/mm3" CD158k+SC/mm3" CARE49 Cl
Slide 15: Analysis of CD158k+ T cells
Analysis of CD158k+ T cells Immunoscope analysis of V families from CD158k+CD4+T cell and CD158k+T cells CD3+CD4+CD158kneg Lymphocytes ARGP38 LESC45 GOLM47 VB17: 0% VB13a: VB06: 22% VB08 16% 12% VB06 CD4 CD158k+SCs CD158k ARGP38 LESC45 GOLM47 VB13a: 21% VB08: 91% VB17: 77% VB06: 96% VB06 Moins-Teis
Slide 16: Phenotypic heterogeneity of CD158k+ T cells: memory and naive subsets
Phenotypic heterogeneity of CD158k+ T cells: memory and naive subsets CD45RA- CD45RA+ CCR7 35% 87% 24% 60% 43% 37% CD27 3 patients at initial diagnosis Moins-Teisserenc et al, J Invest Dermatol, 2015
Slide 17: Phenotypic heterogeneity of T cells
Phenotypic heterogeneity of T cells CD45RA+ • CCR7+CD27= T Naive • CCR7-CD27-= T Effector Terminal CD4 CD45RA- • CCR7+CD27+= T Central Memory • CCR7-CD27+= T Transitional Memory • CCR7-CD27-= T Effector Memory CD3 CD45RA- CD45RA+ Counts CD45RA CCR7 CD45RA TCM TN CCR7 CCR7 TEM TET TTM CD27 CD27
Slide 18: Phenotypic heterogeneity of CTCL
Phenotypic heterogeneity of CTCL • Sezary patients: Central memory T-cells CD45RA-CCR7+CD27+ • Mycosis fungoides: Effector memory T cells CD45RA-CCR7-CD27-
Slide 19: Central memory phenotype
Lymphocyte Lymphocyte s s Vb+ CD4+/- CD158k+ Vb8 CD4 CD158k CD4 SEZ CD158k+ SEZ CD158k+ SEZ SEZ Vb+ Vb+ CD45RA CD45RA CCR7 CD45RA CCR7 CD45RA CD45RA- CD45RA+ CD45RA- CD45RA 41,7 % 38,5 % + CCR7 CCR7 CCR7 CCR7 TCM TN TCM TN TTM TE TET TE TT TET M M M CD27 CD27 CD27 CD27 Phenotype: Central memory : CD
Slide 20: Phenotype Effector Terminal
Phenotype Effector Terminal : CD45RA+CCR7-CD27- SEZ CD158k+ SEZ CD158k+ SEZ Vb+ SEZ Vb+ CD45RA CD45RA CCR7 CD45RA CCR7 CD45RA CD45RA- CD45RA+ CD45RA- CD45RA+ CCR7 CCR7 CCR7 CCR7 TCM TN TN TCM 88,3% 88,8% TEM TTM TET TEM TTM TET CD27 CD27 CD27 CD27
Slide 21: Naive Phenotype
Naive Phenotype : CD45RA+CCR7+CD27+ SEZ CD158k+ SEZ CD158k+ SEZ Vb+ SEZ Vb+ CD45RA CD45RA CCR7 CD45RA CCR7 CD45RA CD45RA- CD45RA+ CD45RA- CD45RA+ 41,7 % 41,7 % CCR7 CCR7 CCR7 CCR7 TN TN TCM TCM TEM TTM TEM TTM TET TET CD27 CD27 CD27 CD27
Slide 22: Flow cytometric analysis of CD3+CD4+KIR3DL2+ cells in erythrodermic mycosis fungoides
Flow cytometric analysis of CD3+CD4+KIR3DL2+ cells in a B1 erythrodermic mycosis fungoides KIR3DL2 34.1% of 88.2% CD3+CD4+ CD3+CD4+ T cells 1a 1b Hurabielle, J Invest Dermatol, 2015
Slide 23: Value of each blood biomarker
Value of each blood biomarker (A.U) K IR 3D 10000 100000 0.1 1 10 100 1000 K L2 IR -B 3D 0 * L2 TW - B IS 1 T TW - B IS 0 * T- T pl - as B1 tin T- pl - B as 0 tin N K -B1 p4 Erythrodermic MF patients 6 N -B K Blood biomarkers in B0 and B1 p4 0 6 -B 1 Hurabielle, J Invest Dermatol, 2015
Slide 24: Sezary cells also express activating Killer cell Ig-like Receptors
Sezary cells also express activating Killer cell Ig-like Receptors
Slide 25: Expression of other KIRs by Sezary cells
Expression of other KIRs by Sezary cells B PBMCs Cell line A 15% 69% 80% V8-C V8-J2.5 V8 V8 CD158k Circulating 9% 7% 1% 19% Sezary cells 280 244 Number of cells Number of cells V8-C CD158a V8-J2.5 Cell line Number of cells Number of cells CD158b 280 244 Marie-Cardine, Blood 2007
Slide 26: What is the function of CD158k/KIR3DL2 on CTCL cells?
What is the function of CD158k/KIR3DL2 on CTCL cells ?
Slide 27: No Lysis
No Lysis Inhibitory jleceptor Lysis Inhibitory Receptor Cc Lysis Inhibitory Activating Receptors tou D No Lysis Inhibitory HLA Receptors Activating Receptor Activating Ligand
Slide 28: Number Number
Prolifération AICD Tumoral T cell clone Non tumoral CD4+ T cells Tumoral T cell clone Non tumoral CD4+ T cells (V9+ KIR3DL2+) (V9- KIR3DL2-) (V14+ KIR3DL2+) (V14- KIR3DL2-) NT 0.81 0.32 0.16 NT 4.31 AZ158 1.5 0.23 0.98 AZ158 4.49 50. 9 CD3 22.1 33.9 91.0 CD3 CD3 21.4 + 2.6 95.8 CD3 28.6 AZ158 +
Slide 29: Inhibition of the CD3-mediated activation upon CD158k recruitment
Inhibition of the CD3-mediated activation upon CD158k recruitment Sézary patient CD4+ cells P1 P2 - + - + IP KIR3DL2 Lysates Blot anti-SHP-1 CD3 CD3 NT CD3 + + mIgG AZ158 Phospho-Erk Erk Phospho- Co-inhibitory receptor
Slide 30: A novel KIR-associated function: CpG DNA uptake mediated by KIR3DL2
A novel KIR-associated function:evidence that CpG DNA uptake and shuttling to early endosomes is mediated by KIR3DL2 Sivori et al, Blood 2010
Slide 31: Visualisation of KIR3DL2-ODN interactions by confocal microscopy
Visualisation of KIR3DL2-ODN interactions by confocal microscopy Co-internalisation of KIR3DL2 and CpG-ODN in endosomes where TLR9 is located Sivori et al, Blood 2010
Slide 32: 7-AAD
LAMP-1 KIR3DL2 Merge NT + ODN-C + Control ODN Day 3 NT ODN-C FITC LAMP-1 Merge ODN-C FITC KIR3DL2 Merge Day 7 + ODN-C ODN-C FITC LAMP-1 Merge -FITC Day 10 7-AAD V3
Slide 33: KIR3DL2-CpG ODN interactions induces Sézary cell death
KIR3DL2-CpG ODN interactions induces Sézary cell death Sézary Patient Gate: TCRV+ CD4+ NT + ODN-C (CpG) + Control ODN 2.6% 18.1% 3.0% Late (7-AADhigh) Early (7-AADlow) 2.7% 9.8% 3.5% 7-AAD FS Ghazi et al, J Invest Dermatol, 2014
Slide 34: What is the distribution of CD158k/KIR3DL2?
What is the distribution of CD158k/KIR3DL2 ?
Slide 35: Sezary Syndrome
Sezary Syndrome
Slide 36: Sezary Syndrome
Sezary Syndrome
Slide 37: Transformed mycosis fungoides
Transformed mycosis fungoides
Slide 38: CTCL landscape: KIR3DL2 expression by WHO-EORTC subtype
CTCL landscape KIR3DL2 expression by WHO-EORTC subtype KIR3DL2 is expressed in ~65% of all CTCL, irrespectively of disease subtype. Expression is more prominent in Sézary syndrome, transformed mycosis fungoides and CD30+ LPD (ALCL subtype)
Slide 39: Primary cutaneous anaplastic large cell lymphoma
Primary cutaneous anaplastic large cell lymphoma
Slide 40: KIR3DL2 expression in primary cutaneous anaplastic large-cell lymphoma
KIR3DL2 expression in primary cutaneous anaplastic large-cell lymphoma
Slide 41: Strong KIR3DL2 expression by skin-infiltrating lymphocytes in pcALCL
Strong KIR3DL2 expression by skin-infiltrating lymphocytes in pcALCL
Slide 42: KIR3DL2 is expressed by Mac2a and Mac2b ALCL lines
KIR3DL2 is expressed by Mac2a and Mac2b ALCL lines • Mac2a and Mac2b cell lines were derived from separate, rapidly growing skin tumors during disease progression, in a pcALCL patient • Mac1 was derived from circulating tumor cells in the blood of the same patient during an earlier indolent course o
Slide 43: Cutaneous g/d T-cell lymphoma
Cutaneous g/d T-cell lymphoma
Slide 44: Extranodal NK/T lymphoma, nasal type
Extranodal NK/T lymphoma,nasal type
Slide 45: Primary cutaneous aggressive epidermotropic CD8+ T-cell lymphoma
Primary cutaneous aggressive epidermotropic CD8+ T-cell lymphoma
Slide 46: Is there a rationale to develop an anti-KIR3DL2 Immunotherapy?
Is there a rationale to develop an anti-KIR3DL2 Immunotherapy ?
Slide 47: NK cells of Sezary patients are functional
NK cells of Sezary patients are functional Bouaziz et al, J Invest Dermatol, 2005
Slide 48: NK lysis of tumor cells is enhanced via an ADCC mechanism
NK lysis of tumor cells is enhanced via an ADCC mechanism Tumoral cell line Pno 70 60 Alemtuzumab 50 40 Tumoral cell line Pno 30 20 10 0 NK + Control Ab NK + Campath Bouaziz et al, J Invest Dermatol, 2005
Slide 49: Sezary patients NK cells are able to degranulate
Sezary patients NK cells are able to degranulate No Target C1R NKG2D expression ratio p < 0.01 50 ns 1,2 ns p < 0.01 % CD107a+ cells p < 0.01 p < 0.01 40 1 CD56 p < 0.01 8.6% 19.9% 30 0,8 0,6 K562 C1R-MICA 20 0,4 10 0,2 0 39.2% 32.2% 0 No Target K562 C1R C1R-MICA No Target K562 C1R C1R-MICA CD107a •
Slide 50: Is it possible to develop a specific anti-KIR3DL2 Immunotherapy?
Is it possible to develop a specific anti-KIR3DL2 Immunotherapy ? Specific expression of KIR3DL2 by malignant T lymphocytes Circulating Sézary patients NK lymphocyte are functional Sézary T cell clones are sensible to Perforin/GrB lysis Development of a therapeutic monoclonal antibody (Innate
Slide 51: Portfolio of anti-KIR3DL2 mAbs
Portfolio of anti-KIR3DL2 mAbs D0 • mAbs binding to epitopes on all 3 Ig domains of KIR3DL2 were generated and all their epitopes were identified • Epitopes (colored AA) bound correlate D1 with distinctive properties (efficient killing of KIR3DL2+ cells, internalization of the molecule, inhibition o
Slide 52: Strategy for generation and testing of anti-KIR3DL2 therapeutic antibodies
Strategy for the generation and testing of anti-KIR3DL2 therapeutic antibodies Immunization of Balb/c mice with KIR3DL2-Fc chimeric protein Selection of KIR3DL2 specific mAb (n = 12) Chimerization Humanization Binding properties Candidate mAb: Functional assays IPH4102 Marie-Cardine A, et al. Cancer
Slide 53: IPH4102 delineates Sézary cells
IPH4102 delineates Sézary cells Patient #17 Patient #21 Patient #20 IPH4102 IPH4102 IPH4102 V22 V1 V2 Marie-Cardine et al, Cancer Res, 2014
Slide 54: In vivo efficacy of IPH4102: experimental design
In vivo efficacy of IPH4102: experimental design i.v 5x106 IPH4102 or IC (i.p.) Raji-KIR3DL2 2 x per week Clinical signs SCID mice and weight Day 1 Day 2 s.c 5x106 IPH4102 or IC Raji-KIR3DL2 2 x per week Tumour size SCID mice and weight Tumour growth Day 1 Day 15–17 (up to 100 mm3) i.p., intraperito
Slide 55: IPH4102 promotes the survival of mice engrafted with KIR3DL2+ tumour cells
IPH4102 promotes the survival of mice engrafted with KIR3DL2+ tumour cells 100 10 µg IC 0.01 µg IPH4102 80 Percent survival 0.1 µg IPH4102 1 µg IPH4102 60 10 µg IPH4102 40 20 0 0 20 30 40 50 60 Days (post cell injection) End of Study Mice: SCID (n = 8) RAJI-KIR3DL2: 5 M IV at D0 IPH4102 improves sur
Slide 56: IPH4102 Efficacy ex vivo: Autologous ADCC
IPH4102 Efficacy ex vivo: Autologous ADCC Efficacy Results Patient #10 Patient #11 Patient #17 Patient #7 100 100 30 50 80 80 40 %7AAD+ CD4+ IPH4102 Sézary cells rituximab 20 60 60 30 alemtuzumab % killing 40 40 20 10 20 20 10 0 0 0 0 E/T ratio IPH4102 as potent as alemtuzumab in ex vivo autologous
Slide 57: KIR3DL2+ Mac2a and Mac2b ALCL lines are sensitive to ADCC mediated by IPH4102
KIR3DL2+ Mac2a and Mac2b ALCL lines are sensitive to ADCC mediated by IPH4102
Slide 58: IPH4102-101 FIH Development
IPH4102-101 FIH Development • Sept 2013-Sept 2014: Preclinical studies • August 2014: Orphan Drug designation in the European Union for the treatment of CTCL • Sept 2014-Sept 2015: Regulatory process • 5 August 2015: ANSM approval for phase I trial in France • 11 Sept 2015: FDA approval • 21 Sept 20
Slide 59: IPH4102-101 Study Design
IPH4102-101 STUDY DESIGN • First-in-Human Phase I study of IPH4102 • Two-portion study: – Dose-escalation portion – 10 dose-levels – Cohort expansion portion, at the recommended dose determined in the dose-escalation – 2 CTCL subtypes (SS and tMF) • Patient profile: – Relapsed/refractory (≥ 2 previo
Slide 60: IPH4102-101 FIH Study Design Objectives
IPH4102-101 FIH Study Design Objectives • Primary objective: to assess safety & tolerability of increasing IV doses of single agent IPH4102 by: • characterizing the dose-limiting toxicities (DLT) and (S)AEs • identifying the MTD or Recommended Ph 2 Dose (RP2D) • Secondary objectives: – To explore an
Slide 61: IPH4102-101 FIH Study Design
IPH4102-101 FIH Study Design Study Design • Cohort expansion Part: same dose for all: RP2D Dose-level pts with KIR3DL2+ tumors 10 pre-selected CTCL subtypes 3 (+3) pts Dose-level 4 Recommended Phase II Dose (RP2D) Dose-level 3 1 (+2+3) pts e.g. n = 10 tMF + 10 SS Dose-level 1 The CTCL subtypes and n
Slide 62: IPH4102-101 FIH Study Design: Participating sites
IPH4102-101 FIH STUDY DESIGN Participating sites Clinical sites (dose-escalation part): – St Louis Hospital, Paris (M. Bagot) – UMC Leiden, the Netherlands (M. Vermeer) – Guy’s and St Thomas’ Hospital, London UK (S. Whittaker) – Stanford U., CA, US (Y. Kim) – MD Anderson, Houston, TX, US (M. Duvic)
Slide 63: Acknowledgements
Innate Pharma (Marseille, France) H. Sicard R. Joly Unit 976 (Paris, France) N. Viaud S. Chanteux A. Bensussan M. Bléry B. Rossi N. Thonnart C. Bonnafous L. Gauthier A. Marie-Cardine C. Paturel L. Michel K. Pilz C. Paiva Dept of Dermatology St Louis Hospital, Paris, France M. Bagot Dept of Immunolog
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