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SFGM-TC, 2018
webvision.med.utah.edu Nei.nih.gov
Christelle Monville, PhD SFGM-TC, 2018 MERTK, RPE65, LRAT - - PowerPoint PPT Presentation
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
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SFGM-TC, 2018
Adapted from Carr et al. TINS 2013
(Thomson et al., 1998) (Takahashi et al., 2007, Yu et al., 2007)
2012 Nobel Prize
John Gurdon Shinya Yamanaka
NOR : AFSB1610277S
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SFGM-TC, 2018
- Cell replacement of the degenerated RPE layer with a RPE layer
generated from human embryonic stem cells (hESCs)
Human embryonic stem cells RPE cells Denuded Human amniotic membrane Photoreceptors Microvilli Nutrients in Waste out Tissue Engineering product (TEP) Brüch’s membrane
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SFGM-TC, 2018
Polarized RPE cells
Non-proliferative Better survival in advanced stages of disease Do not migrate Increased growth factor secretion Better integration
Membrane support
Biological Substrate for RPE to attach & polarize 50-100 mm thickness Non immunogen Anti-inflammatory properties Already used for corneal implantations (ANSM PPT44) Not a limiting source (200-300 ceasarean/year at AP-HP)
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SFGM-TC, 2018
1
hESC-derived RPE bank
Human Amniotic Membrane
Tissue Engineered Product hESCs bank Embryo hESCs derivation and amplification hESCs differentiation
Tissue engineering
Human amniotic membrane bank
Dr Jarraya, Tissue Bank, St Louis, Paris
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SFGM-TC, 2018
RPE Ampli
P1
J117+21
RPE-BANK
P0
RPE Ampli
J94+21
RPE Diff
J0+21 J54+21
hES-RC9 ampli
J0 J21 Pax6 Ezrin MERTK
2 3 4 5 6
h
Basal Apical
VEGF (pg) Weeks 8000 7000 6000 5000 4000 3000 2000 1000 1
- A. Plancheron
- W. Habeler
NOR : AFSB1530503S
BenM’Barek et al. 2017
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SFGM-TC, 2018
2
hESC-derived RPE bank
Human Amniotic Membrane
Tissue Engineered Product hESCs bank Embryo hESCs derivation and amplification hESCs differentiation
Tissue engineering
Human amniotic membrane bank
Dr Jarraya, Tissue Bank, St Louis, Paris
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SFGM-TC, 2018
Amniotic Membrane Cryopreserved AM Denuded AM Thermolysin Already used in clinic
BenM’Barek et al. 2017
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hAM
POS-FITC MERTK/
BenM’Barek et al. 2017
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SFGM-TC, 2018
Daily immunosuppression treatment in the drinking water
- Functional defect of RPE (phagocytosis)
- Photoreceptor death from 21d
- At 60 days postnatal: no ERG response
Photoreceptors alive P14 P30 P105 Delay PR death
Royal college of surgeons (RCS) rat model
- K. Ben M’Barek
BenM’Barek et al., 2017
Use of IOL injectors
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SFGM-TC, 2018
µ µ µ µ µ 9 weeks post-op µ µ µ µ
SHA M RPE suspension TEP-STREAM Dystrophic
µ µ µ
0.25E9 0.5E9 0.75E9 1E9 1.25E9 1.5E9 1.75E9 2E9 2.25E9
µ µ AUC (µV x µcd.s/m^2) µ µ F
**
0 23 23 22 25 24 26 10 19 25 28 32 26 19 0 20 29 0 23 25 28 0 22 28 31 13 23 19 0 25 28 29 28 0 19 26 0 23 28 25 24 22 25 0 38 18 26 25 0 20 20 20 31 26 18 22 20 20 19 32 32 36 32 34 13 15 19 0 22 18 26 23 22 0 19 28 26 22 31 32 12 19 0 25 26 20 32 25 32 32 0 45 37 26 38 31 25 35 0 26 26 19 16 26 20 20 0 25 20 35 19 20 25 0 22 15 0 16 16 15 0 16 23 26 22 22 0 16 22 12 0 18 0 19 23 20 26 32 18 22 20 22 28 22 0 23 29 20 19 13 22 20 25 22 32 23 28 22 23 23 34 31 0 21 18 19 20 20 19 15 16 15 13 18 0 16 15 0 10 18 10 18 16 18 0 16 28 0 19 20 16 12 12 9 18 0 18 10 0 20 18 16 12 20 18 18 19 0 20 10 19 15 18 19 16 16 20 16 19 22 16 16 0 18 18 13 12 18 15 13 13 18 13 9 20 13 13 22 0 16 20 18 15 18 12 0 16 10 18 3 15 13 7 12 7 0 10 15 18 18 10 9 0 13 9 0 12 9 16 18 16 9 10 0 15 10 15 18 15 10 12 19 7 9 12
b
TEP SHAM RPE cell suspension Dystrophic
7 11 10 10 12 0 11 9 8 10 5 5 12 7 12 11 0 12 13 9 10 7 1 1 7 8 8 8 7 9 7 7 5 8 0 11 11 9 18 11 17 10 0 14 0 13 0 15 11 13 0 16 13 13 0 15 7 13 14 0 14 7
50 25
µm
15 10 9 0 10 10 18 7 9 7 9 9 9 9 7 9 7 7 0 13 6 10 0 12 10 12 13 10 9 12 0 16 10 10 0 13 10 6 0 18 9 22 12 9 7 10 13 18 12
a’ b’ µ Width (µm)
*
Width (µm) µ µ
Temporal Nasal Dorsal Ventral Temporal Nasal Dorsal Ventral Temporal Nasal Dorsal Ventral
BenM’Barek et al., 2017
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12 therapeutic units GMP facility 6 primates Females 5/8 years old; immunosuppression Baseline: ERG, OCT fundus Surgery D1/2 W1 W2 W4 W6 W7 Baseline: ERG, necropsy, Histology of the retina Slit lamp, fundus, OCT
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Patent: Ben M’Barek et al.
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B a s e l i n e T r a n s p l a n t e d 1 0 2 0 3 0 4 0 5 0
B w a v e _ im p lic ite tim e B L v s T R
m s
n s B a s e lin e T ra n s p la n te d 50 100 150 200
B w av e am p litu d e B L v s T R
µ V
n s
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▪ 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)
- f
the Tissue engineered product (TEP) in animal models (mouse, rats and Monkeys) ▪ Clinical trial planned in 2019 in patients with MERTK, RPE65, LRAT mutations
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DAPI / MITF / ZO1 DAPI / TYRP1/ PAX6
hESCs RPEs P1
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SFGM-TC, 2018
▪ Karim Ben M’Barek ▪ Walter Habeler ▪ Alexandra Plancheron ▪ Florian Regent ▪ Marc Peschanski Preclinical development ▪ Thierry Bordet ▪ Ana Bejanariu Clinical development ▪ Géraldine Honnet ▪ Florence Ganne Regulatory affairs
- Didier Caizergues
- Helene Lafolly