T-cell engineering for adoptive immunotherapy using TALeffector. Sophie Derniame PhD Project Leader, CellectisTherapeutics

Transcription

1 T-cell engineering for adoptive immunotherapy using TALeffector nucleases (TALEN ) Sophie Derniame PhD Project Leader, CellectisTherapeutics

2 FORWARD LOOKING STATEMENT This communication expressly or implicitly contains certain forward-looking statements concerning Cellectis SA and its business. Such statements involve certain known and unknown risks, uncertainties and other factors, which could cause the actual results, financial condition, performance or achievements of Cellectis SA to be materially different from any future results, performance or achievements expressed or implied by such forward-looking statements. Cellectis SA is providing this communication as of this date and does not undertake to update any forward-looking statements contained herein as a result of new information, future events or otherwise. Cellectis proprietary information. Not to be copied, distributed or used without Cellectis s prior written consent. 2

3 Retargeting T-cells to tumors with CARs Chimeric Antigen Receptors (CARs) Tumor cell CARs = cell membrane bound mabs Tumor antigen binding: mab scfv Allows MHC-independent antigen recognition T-cells replicate in response to contact with antigen in vivo T-cell Adoptive autologous T-cell immunotherapy using CARs directed at CD19 has shown impressive results with complete remissions of late stage leukemias 3

4 Universal anti-cd19+ adoptive T-cell immunotherapy CAR expression to redirect T-cells to tumor antigens Suicide gene for safety Off the shelf (TCR disruption*) Engineered SG SG Minimal potential for GvHD For CLL - CD52 disruption* to prevent destruction by high risk CLL mab Alemtuzumab therapy (*): knock-out by using TALEN mrna Allogeneic T-cell therapy using cells derived from healthy donors 4

5 Platform development: combining multiple technologies 17 Days CAMPATH conditioning (non-cytotoxic) Apheresis (Frozen PBMC) T-cell activation Stable gene transfer Gene KO (transient nuclease expression) T-cell expansion and purification Product storage Patients 5

6 % cell lysis % cell lysis Anti-Fab (scfv) CAR transduction UCART19 CAR exhibits comparable/higher cytolytic capability than benchmark CAR on target cells SUPT1 CD19 negative cells 30:1 15:1 8:1 3:1 NT supe Benchmark CAR MP13622 supe Ctx CAR MP13624 Effector:Target SUPT1-CD19 CD19 expressing cells NT supe Benchmark CAR MP13622 supe Ctx CAR MP :1 15:1 8:1 3:1 Effector:Target In collaboration with Dr. Martin Pule, UCL, UK 6

7 SSC Viability Cell count CTx proprietary technology for TALEN electroporation AgilePulse TM Electroporation system Large volume chamber ( ml) Electroporation buffer A series of high voltage, short duration pulses followed by lower voltage, longer duration pulses 93% GFP+ High transfection efficacy High cell viability FSC GFP GFP 7

8 TCR/CD3 TALEN -mediated knock-outs TCR CD % CD52 TALEN mrna: transient gene transfer FCS 72.9 % FCS 51.1% Functional tests: TCR KO cells are unresponsive to TCR stimulation CD52 KO cells are resistant to Campath depletion 8

9 CD45RA CD4 TCR/CD3 End product Cell expansion around 80 fold in 11 days (5 x 10 6 to 378 x 10 6 ± 54 cells) Depletion of TCRαβ+ cells (Miltenyi) Final phenotype (day 17): CD8 CD52 Starting population (CD3 + ) Final population (CD3 - ) Before purification After purification CD62L 9

10 CAR19 TCR - cells eradicate lymphoma in an orthotopic model No CAR CAR19 T-cell CAR19 TCR - T-cell Day 2 Day 1: iv infusion with 250k Raji Day 2: iv infusion with 4x10 6 T-cells Day 7 TCR-deficient cells engraft as WT T-cells TCR-deficient cells eradicate Raji lymphoma Day 12 In collaboration with Dr. Martin Pule, UCL, UK 10

11 Cellectis established manufacturing process platform A robust process designed for cgmp compatibility 10 9 T-cells 2x T-cells D0 D3 D5 D17 Healthy Donor Apheresis Frozen PBMC Activation T-cells Transduction Efficiency >50% Electroporation TALEN mrna Double KO Efficiency >40% Cell Separation 1 x CAR + TCR - T-cells >40% CD52 KO Patients Off the shelf cell therapy product 11

12 Conclusion Development of an off-the-shelf therapeutic T-cell platform Efficient T-cell engineering for exogenous CAR expression TALENs permit efficient and simultaneous elimination of Endogenous T-cell receptor significant decrease of GvHD risk Cellular target for a lymphodepleting treatment for proliferation & activity of final T-cells versus immunosuppressive drugs A robust process designed for cgmp compatibility Generation of non alloreactive T-cells for cancer therapy 12

13 Acknowledgments Cellectis therapeutics Julianne Smith Andrew Scharenberg Cellectis Platform Philippe Duchateau Jean-Charles Epinat Collaborators: Laurent Poirot Roman Galetto Sylvain Arnould Agnès Gouble Cécile Schiffer- Mannioui Stéphanie Grosse University College, London Martin Pule - Brian Philip - Gordon Weng-Kit Cheung - Leila Mekkaoui Karl Peggs Waseem Qasim Isabelle Chion Diane LeClerre Céline Lebuhotel Justin Eyquem Laetitia Lemaire Pierrick Potrel Cécile Bas 13

14 THANK YOU