Creating a New Class of Durable Redosable Gene Therapy - FOR MILLIONS OF PATIENTS LIVING WITH RARE AND PREVALENT DISEASES
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Creating a New Class
of Durable Redosable
Gene Therapy
FOR MILLIONS OF PATIENTS LIVING
WITH RARE AND PREVALENT DISEASES
NASDAQ: GBIO MAY 2021
Non-Viral Gene Delivery
of Human FVIII to
Hemophilia A Mice and
Non-Human Primates
MAT THEW G. STANTON , DEB KLAT TE, MAT T CHIOCCO,
LUKE HAMM, ELIZABETH NELSON, LEAH LU, GREG
FEINSTEIN, NOLAN GALLAGHER, JIE SU, ANDREW
MILSTEAD, DI BUSH, RUSSELL MONDS, NICHOLAS
PARSONNET, ASHLEY PENVOSE, SARAH LAGOY, ERIK
HANSEN, JONATHAN KIT TEL , ANASTASIA LYMAR, KATI
VU, MICHELLE RODRIGUEZ JOYCE, KARL MALAKIAN,
JEFF MOFFIT T
MAY 2021
Forward-looking statements Any statements in this presentation about future expectations, plans and prospects for the company, including statements about our strategic plans or objectives, our technology platforms, our research and clinical development plans, and other statements containing the words “believes,” “anticipates,” “plans,” “expects,” and similar expressions, constitute forward-looking statements within the meaning of The Private Securities Litigation Reform Act of 1995. Actual results may differ materially from those indicated by such forward-looking statements as a result of various important factors, including: uncertainties inherent in the identification and development of product candidates, including the conduct of research activities, the initiation and completion of preclinical studies and clinical trials and clinical development of the company’s product candidates; uncertainties as to the availability and timing of results from preclinical studies and clinical trials; whether results from preclinical studies will be predictive of the results of later preclinical studies and clinical trials; expectations for regulatory approvals to conduct trials or to market products; challenges in the manufacture of genetic medicine products; whether the company’s cash resources are sufficient to fund the company’s operating expenses and capital expenditure requirements for the period anticipated; the impact of the COVID-19 pandemic on the company’s business and operations; as well as the other risks and uncertainties set forth in the “Risk Factors” section of our most recent annual report on Form 10-K, which is on file with the Securities and Exchange Commission, and in subsequent filings the company may make with the Securities and Exchange Commission. In addition, the forward-looking statements included in this presentation represent the company’s views as of the date hereof. The company anticipates that subsequent events and developments will cause the company’s views to change. However, while the company may elect to update these forward-looking statements at some point in the future, the company specifically disclaims any obligation to do so. These forward-looking statements should not be relied upon as representing the company’s views as of any date subsequent to the date on which they were made. 3
Generation Bio has built the foundation to unlock the full potential of gene therapy
Non-immunogenic Large cargo capacity
• Patients with existing antibodies to AAV • Genes that are too large for AAV
• Titration to effect for every patient • More potent constructs
ceDNA
PATIENTS • Ability to intervene early in life when it closed-ended DNA • Addition of regulatory elements
matters most
Manufacturing scale & speed Specific, on-target delivery
• Rapid cycle from idea to test article • High selectivity for cell type of choice
(4 weeks)
• Amenable to many tissues/cell types
• Currently comparable scale and cost
RAPID to biologics ctLNP
DEVELOPMENT &
SCALING ENGINE cell-targeted LNP
• Move to enzymatic process enables
scale more comparable to mRNA
4
To unlock this potential, solutions to two independent, decades old challenges are
required
Innate Immunity Access to Nucleus
• Non-viral delivery systems deliver DNA to endosome • Non-viral delivery systems such as LNPs only deliver
and cytoplasm of macrophages which are rich in DNA cargo to the cytoplasm of target cells
pattern recognition receptors (PRRs)
• For gene therapy applications, independent solutions
• AAV “shields” DNA from innate immune sensors until DNA
is in nucleus to nuclear uptake must be implemented
IFN-alpha
IFN-gamma pDNA injected into pDNA injected into
1500
control 400
control nucleus: cytoplasm:
Mean Concentration
Mean Concentration
standard LNP standard LNP
300
1000
(pg/ml)
(pg/ml)
200
500
100
0 0
• This response limits doses of traditional LNPs/DNA to
0.5 mg/kg or less – below where these systems can plasmid plasmid
drive therapeutically meaningful expression
100% of cells GFP positive 0% of cells GFP positive
5
We have developed unique and proprietary solutions to both challenges
Innate Immunity Access to Nucleus
• By engineering our LNPs to “de-target” macrophages • Unlike pDNA, our closed-ended DNA construct
and then actively re-targeting them to desired cell provides GFP positive cell populations after delivery to
types (hepatocytes in liver) we have dramatically the cytoplasm of cells
opened the therapeutic index for non-viral DNA
delivery. We call these cell-targeted LNPs or ctLNP
ceDNA injected into ceDNA injected into
• 0.5 mg/kg; 6 hours post dose serum IFNa: nucleus: cytoplasm:
IFN-alpha
2000
PBS
Mean Concentration
standard LNP
1500
ctLNP
(pg/ml)
1000
500 ceDNA
0 100% of cells GFP positive 40% of cells GFP positive
6
More context on ctLNP – our strategy critically relied on the ability to eliminate
phagocytosis by macrophages while maintaining endosomal escape
Phagocytosis assay (labeled LNP pH dependent endosomal escape Addition of GalNAc to target
uptake into human macrophages) assay hepatocytes (6 hr qPCR data)
• LNPs are labelled with DiD • LNPs are incubated with • Perfused tissues were
serum and followed by mixing collected at 6 hours and
• THP1 monocytes are
with endosome levels of ceDNA were
differentiated into
mimicking liposomes for a determined by qPCR
macrophages
set period of time at 37°C
• Macrophages are incubated
• The incubation occurs at
with LNPs for a set period of
controlled pHs
time, cells are then washed
and imaged for visualization • DNA release from LNPs is
measured using PicoGreen
assay
• Adapted from Zhang et al.,
Journal of Controlled Release
174 (2014) 7–14.
7More context on ctLNP – our strategy critically relied on the ability to eliminate
phagocytosis by macrophages while maintaining endosomal escape
Phagocytosis assay (labeled LNP pH dependent endosomal Addition of GalNAc to target
uptake into human macrophages) escape assay hepatocytes (6 hr qPCR data)
Standard LNPs Standard LNPs Endosomal Release Standard LNPs
ceDNA copies per organ
50% pH 6.0
1 x 1010
Free ceDNA
40%
8 x 109
30% 6 x 109
20% 4 x 109
10% pH 7.4 2 x 109
0
0%
Spleen Liver
Base
ctLNP ctLNP Endosomal Release ctLNP
ceDNA copies per organ
50% 2.5 x 1011
pH 6.0
Free ceDNA
40% 2 x 1011
30% 1.5 x 1011
20% 1 x 1011
10% pH 7.4 5 x 1010
0% 0
Novel Spleen Liver
8Solutions to innate immunity and/or nuclear uptake open significant new and differentiated
opportunities
Rare liver diseases
Lead programs in Hemophilia A and PKU with
rapid follow on in Wilson disease, Gaucher and
others
See Monds poster
Gene editing in rare liver and retina
Targeted gene insertion for one-time correction Retinal diseases
Lead programs in LCA10, Stargardt and Wet
AMD
ceDNA ctLNP CMC
Vaccines
Antibody gene transfer
Examples include hepatitis B, HIV and sickle cell
disease, Wet AMD, etc.
See Silver poster
Targeted therapy across multiple tissues
Examples include skeletal muscle, CNS and
oncology therapies
9
Solid arrows = focus areas; spotted gray arrows = conceptualSolutions to innate immunity and/or nuclear uptake open significant new and differentiated
opportunities
Rare liver diseases
Lead programs in Hemophilia A and PKU with
rapid follow on in Wilson disease, Gaucher and
others
See Monds poster
Gene editing in rare liver and retina
Targeted gene insertion for one-time correction Retinal diseases
Lead programs in LCA10, Stargardt and Wet
AMD
ceDNA ctLNP CMC
Vaccines
Antibody gene transfer
Examples include hepatitis B, HIV and sickle cell
disease, Wet AMD, etc.
See Silver poster
Targeted therapy across multiple tissues
Examples include skeletal muscle, CNS and
oncology therapies
10
Solid arrows = focus areas; spotted gray arrows = conceptualHemophilia A
Redosable childhood intervention; titration to correct level in each patient
D I S E A S E O U R A P P R O A C H
~16,000 patients in US
Diagnosis in early childhood
Bleeding disorder caused by
deficiency in clotting factor VIII
Gene therapy in development
limited to adults, efficacy is
variable and waning
P O T E N T I A L T H E R A P E U T I C O P P O R T U N I T Y
★ Greater potency based on larger genetic payload
★ Titration to target expression level for each patient
★ Dosing in childhood before disease progression
★ Redosing to extend benefit over a lifetime
Atalay et al., Ital. J. Med. 9(3): 290-293 (2015)
AAAS EurekAlert! 30 Aug 2018 Univ. Witwatersrand
11Demonstrated durability and redosing with Factor IX in immune competent mice
No decrease in expression throughout study period, redosing increases expression proportionately
Durability FIX used as surrogate for durability and redosing in wildtype mice because this human
& redosing protein does not raise neutralizing antibodies in mice, unlike human FVIII
in mice
Factor IX Factor IX
100
FIX (% WT activity)
10
1
0.1
0 7 14 21 28 35 42 49 56 63 70 77
Day
• Single IV administration at study day 0 • Single IV administration at study day 0
• Factor IX activity calculated from protein ELISA • Re-dosed at day 36 at same dose level
• Factor IX activity calculated from protein ELISA
12Mouse proof of concept and translation of expression from mouse to NHP
Target levels of factor VIII expression & Translation of expression across species
in Hemophilia A mice from mouse to NHP
Development Construct: FVIII Expression Research Construct: Species Translation
Systemic IV administration via ctLNP (Day 10) Systemic IV administration via ctLNP (day 5 or 7)
PBS Study 1: ctLNP1 Study 2: ctLNP2
100 @ 1 mg/kg @ 2 mg/kg
GB1
4 4
40
(% normal)
3 3
30
% normal
% normal
(% normal)
10 Target
FVIII
FVIII
FVIII
FVIII
2 2 20
1 1 10
0
1 0 0 0.0 0.5 1.0 2
)
)
)
)
)
)
Dose (mg/kg
=2
=2
=4
=5
=4
=2
(n
(n
(n
(n
n
n
P,
P,
0.0 0.5 1.0 2.0
P
P
se
se
H
H
H
H
ou
ou
N
N
N
N
(
(
M
M
Dose (mg/kg)
ol
ol
tr
tr
on
on
C
C
13Summary • Non-viral delivery of DNA represents enormous opportunity to expand reach of genetic medicines (gene transfer and gain of function gene editing) • Two decades-old hurdles have been uniquely overcome at Generation Bio – ctLNP to dampen innate immune stimulation and ceDNA to gain access to target cell nuclei • Durable expression of non-immunogenic protein demonstrated in wildtype mice • Redosing of non-immunogenic protein demonstrated in wildtype mice • Expression of FVIII for hemophilia A at therapeutically meaningful levels has been achieved in mice • A roughly 2:1 species translation of expression between mouse and non-human primates has been achieved with tolerability in NHP up to 2 mg/kg 14
Thank you! 15
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