International Regulatory Forum of Human Cell Therapy and Gene Therapy Products Ian Harris, Ph.D. 16 March 2016 Cell Therapy, Janssen R&D Preclinical proof of concept and other preclinical issues Jennifer Jacobs, Stowaway Jennifer is a New York based artist living with Type 1 diabetes.
27
Embed
Preclinical proof of concept and other preclinical issues · Preclinical proof of concept and other preclinical issues . Jennifer Jacobs,Stowaway Jennifer is a New York based artist
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
International Regulatory Forum of Human Cell Therapy and Gene Therapy Products
Ian Harris, Ph.D. 16 March 2016 Cell Therapy, Janssen R&D
Preclinical proof of concept and other preclinical issues
Jennifer Jacobs, Stowaway Jennifer is a New York based artist
living with Type 1 diabetes.
Proof of concept (POC)
Scientific Rational for clinical Biologically active dose levels Dosing regimens (single, repeated) Appropriate route of administration Duration of response Mechanism of action
– Protection/prevention – Regeneration
Timing of administration relative to disease/injury Potential patient population Monitoring efficacy and safety
3
Umbilical Tissue Derived Cells (palucorcel, CNTO 2476, hUTC) for the treatment of Geographic Atrophy
• Tumorgenicity • Immunogenicity • Safety of delivery
ICH S6(R1) Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals FDA Guidance for Industry: Preclinical Assessment of Investigational Cellular and Gene Therapy Products (November 2013)
Royal College of Surgeons (RCS) rat
• The RCS rat is a widely studied animal model of retinal degeneration due to a genetic defect in the retinal pigmented epithelium (RPE), i.e., a null mutation of the gene encoding Mertk.
• Cell culture studies demonstrate that Mertk-deficient primary RCS RPE cells have normal levels of outer segments binding, but are defective in the ingestion phase of phagocytosis.
• Delivery of a recombinant Mertk to RCS RPE completely corrected the phagocytic defect of the cells, resulting in photoreceptor rescue.
Palucorcel (hUTC) preserves retinal architecture
Untreated Untreated
CNTO 2476 CNTO 2476
Postnatal d30 Postnatal d90
Sham Sham
• Single injection of 20,000 human cells at Postnatal day 21 in RCS rat preserves photoreceptors to postnatal day 90
Lund RD, et al., Cells isolated from umbilical cord tissue rescue photoreceptors and visual functions in a rodent model of retinal disease. Stem Cells. 2007 Mar;25(3):602-11.
15
Area of Photoreceptor Preservation in the RCS rat
Injection site
Superior Inferior
Outer nuclear layer
Area preserved >30% rat retina
Photoreceptors are preserved beyond where CNTO 2476 is injected
Palucorcel (hUTC) Improves Optomoter Response over 180 d
Optomotor response (head tracking) is used to measure the elicitation of a consecutive motor response, enabling assessment of all components of visual integration. Animals placed in a rotating drum covered with vertical black and white stripes at various spatial frequencies follow the movement of the drum by moving the eye/head whereas animals with altered vision cannot track the stripes
Optomotor Response (6 mo)
Days Postnatal V
isua
l Acu
ity (c
/d)
0 20 40 60 80 100 120 140 160 180 200
0.0
0.1
0.2
0.3
0.4
0.5
Untreated (n=26)
Sham (n=12)
Subretinal (n=13)
Injection P21
Functionally Blind
Normal
* (p<0.001) CNTO 2476 sham untreated
Normal
0 10 20 30 40 50 60 70 80 90 100 0.0
0.1
0.2
0.3
0.4
0.5
0.6
Days Postnatal
Vis
ual A
cuity
(c/d
)
Optomotor Response (3 mo)
Functionally Blind
(n=7) (n=5) (n=3)
Correlation of luminescence with anatomical preservation
Luminance threshold is used to measure spatial vision as single and multiunit activity in the superficial layers of the superior colliculus
Log Candela Units/m2 Subretinal
No sham response
CNTO 2476
Postnatal day 200
0
20
40
60
80
100
0.7 1.2 1.7 2.2 2.6 3.1 3.6 4.1
CNTO 2476 sham untreated
*
* * *
% A
rea
Postnatal day 100
Log Candela Units/m2 Subretinal
Larger area of retina is sensitive to lower levels of light with treatment
0.10.150.2
0.250.3
0.350.4
0.450.5
0.550.6
P20 P30 P60 P75 P90
thre
shol
d (c
/d)
WT
Dys UNT
Sham
4K hUTC
20K hUTC
100KhUTC
Optomoter does not show a dose response
P21 Injection
Confidential
4,000 hUTC 20,000 hUTC 100,000 hUTC
Sham Untreated
Wild Type
19
hUTC was detected in the RCS rat eyes 60 days post subretinal injection
• In order to understand cell fate after subretinal injection, biodistribution studies were performed based on an indirect measurement of human gene expression in animal tissue using Q-RT-PCR
• Biodistribution studies demonstrated long-term cell retention of hUTC in eyes
• hUTC were detected in nontarget organs (peripheral blood, kidney and liver) in nude rats 2 weeks following subretinal delivery, but there was no evidence for long term survival of cells in non-target organs
Human cell retention in the rat eye and visual function
0 25 50 75 100 125 150 1750
1
2
3
4
5
Log cell numberoptomotor threshold recording
0.0
0.1
0.2
0.3
0.4
0.5
Slope -0.01909 ± 0.003782
Slope -0.002198 ± 0.0003982
day
Log
cell
num
ber
optomotor threshold recording
• Similar rate of change are observed in cell number and Optomoter • Optomoter response starts to decline when estimated ~500 cells remain
21
Administration
• Intravitreal • Choroid • Subretinal
Photoreceptors
targeted layer for cell delivery Retinal Pigment Epithelium
Bruch’s Membrane
Choroid
X X
Sclera Choroid
Retina
22
Development of surgical procedure & devices
Transvitreal Suprachoroidal
RCS-RPE
hUTC
In vitro Assay Trophic Factor RCS Rat Model
(RCS phagocytosis studies performed in collaboration with Dr. George Inana, Bascom Palmer Eye Institute)
020406080
100120140160
Normal Dystrophic Dystrophic + hUTC
Rel
ative
pha
gocy
tosi
s *
0
20
40
60
80
100
120
140
N+control M D+control M D+hUTC CM
Rel
ativ
e ph
agoc
ytos
is
*
RCS-RPE
hUTC CM
From preclinical proof of concept to potency assay
24
Hypothesis: Palucorcel rescues phagocytosis in RCS RPE through the secretion of Receptor tyrosine kinase (RTK) ligands
Receptor tyrosine kinase ligands can compensate for loss of Mertk function
BDNF HGF GDNF
Cao, Jet al., (2015) Human umbilical tissue-derived cells rescue retinal pigment epithelium dysfunction in retinal degeneration Stem Cells. doi: 10.1002/stem.2239. [Epub ahead of print]
Receptor kinase ligands secreted by umbilical tissue derived cells are required for bioactivity
Cao, Jet al., (2015) Human umbilical tissue-derived cells rescue retinal pigment epithelium dysfunction in retinal degeneration Stem Cells. doi: 10.1002/stem.2239. [Epub ahead of print]