Christelle Monville, PhD SFGM-TC, 2018 MERTK, RPE65, LRAT - PowerPoint PPT Presentation

Christelle Monville, PhD SFGM-TC, 2018

MERTK, RPE65, LRAT webvision.med.utah.edu SFGM-TC, 2018 Nei.nih.gov

(Thomson et al., 1998) NOR : AFSB1610277S 2012 Nobel Prize (Takahashi et al., 2007, Yu et al., 2007) John Gurdon Shinya Yamanaka SFGM-TC, 2018 Adapted from Carr et al. TINS 2013

• Cell replacement of the degenerated RPE layer with a RPE layer generated from human embryonic stem cells (hESCs) Human embryonic stem cells Photoreceptors RPE cells Microvilli Denuded Human amniotic membrane Brüch’s membrane Nutrients in Waste out Tissue Engineering product (TEP) SFGM-TC, 2018

Polarized RPE cells Membrane support  Biological Substrate for RPE to attach &  Non-proliferative polarize  Better survival in advanced stages of  50-100 m m thickness disease  Non immunogen  Do not migrate  Anti-inflammatory properties  Increased growth factor secretion  Already used for corneal implantations  Better integration (ANSM PPT44)  Not a limiting source (200-300 ceasarean/year at AP-HP) SFGM-TC, 2018

1 hESCs derivation hESCs differentiation and amplification Embryo hESCs bank hESC-derived RPE bank Tissue Engineered Product Tissue engineering Human Amniotic Membrane Human amniotic membrane bank Dr Jarraya, Tissue Bank, St Louis, Paris SFGM-TC, 2018

A. Plancheron W. Habeler J0 J21 J0 +21 J54 +21 J117 +21 J94 +21 hES-RC9 ampli RPE Diff RPE Ampli RPE Ampli P1 P0 RPE-BANK NOR : AFSB1530503S h Pax6 Ezrin MERTK 8000 Apical 7000 Basal 6000 VEGF (pg) 5000 4000 3000 2000 1000 0 1 2 3 4 5 6 Weeks SFGM-TC, 2018 BenM’Barek et al. 2017

hESCs derivation hESCs differentiation and amplification Embryo hESCs bank hESC-derived RPE bank 2 Tissue Engineered Product Tissue engineering Human Amniotic Membrane Human amniotic membrane bank Dr Jarraya, Tissue Bank, St Louis, Paris SFGM-TC, 2018

Already used in clinic Amniotic Membrane Cryopreserved AM Denuded AM Thermolysin SFGM-TC, 2018 BenM’Barek et al. 2017

MERTK/ POS-FITC hAM SFGM-TC, 2018 BenM’Barek et al. 2017

Royal college of surgeons (RCS) rat model K. Ben M’ Barek - Functional defect of RPE (phagocytosis) - Photoreceptor death from 21d - At 60 days postnatal: no ERG response Photoreceptors alive Delay PR death P30 P14 P105 Daily immunosuppression treatment in the drinking water Use of IOL injectors BenM’Barek et al., 2017 SFGM-TC, 2018

µ µ µ µ µ 9 weeks post-op Dystrophic µ TEP RPE cell suspension SHAM Dystrophic µ b ’ TEP-STREAM Dorsal Dorsal Dorsal 0 7 11 10 10 12 0 0 0 0 11 9 8 10 5 5 12 7 12 11 0 0 0 0 23 23 22 25 24 26 10 19 25 28 32 26 19 0 0 0 15 10 9 0 0 0 10 10 18 7 9 0 0 0 0 0 0 0 0 0 RPE suspension 0 0 12 13 9 10 7 1 0 1 7 8 8 8 7 9 7 7 5 8 0 0 0 0 0 0 0 20 29 0 23 25 28 0 22 28 31 13 23 19 0 7 9 9 9 0 9 7 9 7 7 0 0 0 0 0 0 0 0 0 SHA M 0 0 0 0 0 0 0 0 0 0 0 0 11 11 9 18 11 17 10 0 0 25 28 29 28 0 19 26 0 23 28 25 24 22 25 0 38 18 26 25 0 0 0 13 6 10 0 12 10 12 13 10 0 0 9 12 0 0 0 0 0 0 0 0 14 0 0 13 0 15 11 13 0 0 0 0 0 0 0 0 0 0 20 20 20 31 26 18 22 20 20 19 32 32 36 32 34 13 15 19 0 0 0 0 0 0 16 10 10 0 0 0 13 10 6 0 0 0 0 0 0 0 0 0 0 0 16 13 13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 22 18 26 23 22 0 0 0 19 28 26 22 31 32 12 19 0 0 0 0 0 0 0 18 9 22 12 0 0 0 0 9 0 0 0 0 0 a ’ µ m 0 0 0 0 0 0 0 0 0 0 0 0 15 7 13 14 0 0 0 0 0 0 25 26 20 32 25 32 32 0 0 0 45 37 26 38 31 25 35 0 7 10 13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 F ** 0 0 0 14 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 26 26 19 16 26 20 20 0 0 0 25 20 35 19 20 25 0 18 12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2.25E9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 22 15 0 0 16 16 15 0 16 23 26 22 22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Temporal Temporal Nasal Temporal 50 µ Nasal 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 16 22 12 0 18 0 19 23 20 26 32 18 22 20 22 28 22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Nasal 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 23 29 20 19 13 22 20 25 22 32 23 28 22 23 23 34 31 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2E9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 21 18 19 20 20 19 15 16 15 13 18 0 0 16 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 10 18 10 18 16 18 0 16 28 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 19 20 16 12 12 9 18 0 0 18 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 µ ( µ V x µ cd.s/m^2) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 20 18 16 12 20 18 18 19 0 20 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1.75E9 µ 25 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 19 15 18 19 16 16 20 16 19 22 16 16 0 18 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 18 13 12 18 15 13 13 18 13 9 20 13 13 22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 16 20 18 15 18 12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1.5E9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 16 10 18 3 15 13 0 7 12 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 10 15 18 18 10 0 0 9 0 13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9 0 0 12 9 16 18 16 9 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1.25E9 AUC 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15 10 15 18 15 10 12 19 7 9 12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ventral Ventral Ventral 1E9 b * 0.75E9 0.5E9 µ 0.25E9 µ 0 µ µ µ Width (µm) µ µ µ µ BenM’Barek et al., 2017 SFGM-TC, 2018 Width (µm)

Baseline: ERG, OCT Surgery fundus 6 primates Females 5/8 years old; immunosuppression D1/2 W1 W2 W4 W6 W7 Baseline: Slit lamp, fundus, OCT ERG, necropsy, Histology of the GMP facility retina 12 therapeutic units SFGM-TC, 2018

SFGM-TC, 2018 Patent: Ben M’Barek et al.

SFGM-TC, 2018

B w av e am p litu d e B L v s T R B w a v e _ im p lic ite tim e B L v s T R 200 5 0 n s 4 0 n s 150 3 0 µ V m s 100 2 0 50 1 0 0 0 B a s e lin e T ra n s p la n te d e d n e i t l n e a s l a p B s n a SFGM-TC, 2018 r T

SFGM-TC, 2018

▪ We have developed a protocol to generate an unlimited number of clinical grade RPE from hESCs (GMP bank ready). ▪ In vitro validation of hAM as a support allowing the formation of an epithelial sheet of RPE suitable for clinical application. ▪ Functionality/Safety (preclinical) of the Tissue engineered product (TEP) in animal models (mouse, rats and Monkeys) ▪ Clinical trial planned in 2019 in patients with MERTK, RPE65, LRAT mutations SFGM-TC, 2018

DAPI / MITF / ZO1 DAPI / TYRP1/ PAX6 hESCs RPEs P1 SFGM-TC, 2018

▪ Karim Ben M’Barek ▪ Olivier Goureau ▪ ▪ Walter Habeler José-Alain Sahel ▪ Sophie Derenne ▪ ▪ Alexandra Plancheron Céline Nouvel-Jaillard ▪ Thomas Zuliani ▪ ▪ Florian Regent Isabelle Audo ▪ Pauline Souetre ▪ ▪ Marc Peschanski Elena Brazhnikova ▪ Anne-Gaelle Chartois Preclinical development ▪ ▪ Thierry Bordet Stéphane Bertin ▪ ▪ Ana Bejanariu Saddek Mohand Said ▪ Mohammed Jarraya Clinical development ▪ Jerome Larghero ▪ Géraldine Honnet ▪ Valérie Vanneaux ▪ Florence Ganne ▪ Cyrille Maréchal Regulatory affairs ▪ • Nadège Morand ▪ Didier Caizergues Claire-Maelle Fovet • Helene Lafolly ▪ Paul de Sousa ▪ Janet Downie SFGM-TC, 2018