Post on 10-Mar-2020
transcript
Jennifer J. Moseley1, Saravana Kanagavelu1, Hocine Rachid2, Chris Sakoda1, Liang Li1, Sharon Vaturi1
Mario Fournier3, Rachel Smith1, Linda Marban1, Luis Rodriguez-Borlado1
1Capricor Therapeutics, 8700 Beverly Boulevard, Davis Building, Los Angeles, CA;2HLA et Medicine, Hopital Saint Louis, Paris, France; 3Cedars-Sinai Medical Center, Los Angeles, CA
CONCLUSIONS
REFERENCES
1. Blake, D., Weir, A., Newey, S., Davies, K. (2002) Function and
Genetics of Dystrophin and Dystrophin-Related Proteins in
Muscle. Physiological Reviews 82, 2, 291-329.
2. Malliaras, K. Makkar, R. R., Smith, R. R., Cheng, K., Wu, E.,
Bonow, R. O., Marban, L., Mendizabal, A., Cingolani, E.,
Johnston, P. V., Gerstenblith, G., Schuleri, K. H., Lardo, A. C.,
and Marban, E. (2014) Intracoronary Cardiosphere-Derived
Cells After Myocardial Infarction. J. Am. Coll. Cardiol. 63, 110-
122.
UK NMTRC 2018
BACKGROUND
Duchenne Muscular Dystrophy (DMD) is Associated with
Chronic Inflammation
Figure 1. Dystrophin (green) is a large protein that anchors the
sarcolemma to the actin cytoskeleton and has an important
structural role during muscle contraction and relaxation1.
RESULTS
Systemic Therapeutic Effects with IC Delivery in
HOPE Trial
ABSTRACT
Allogeneic CDCs have a low immunogenic profile and repeat-dosing showed additional improvement
when compared with single treatment in a Duchenne muscular dystrophy model
Recently, positive systemic skeletal muscle effects
were reported in the HOPE I clinical trial (Figure 4). The
positive results observed in HOPE on both cardiac and
skeletal muscle function motivated additional studies
exploring new delivery routes and regimens that can
maximize therapeutic benefit of CDCs, allow for
multiple administrations, and reduce patient
administration discomfort.
We analyzed the therapeutic activity of allo-CDCs
delivered intravenously (jugular) into mdx mice.
Improvement in exercise capacity was measured after
the first and second allo-CDC dose, 6 weeks apart
(Figure 5A). Analysis of serum alloantibodies reveals
weak production of alloantibodies against the allo-
CDCs (Figure 5B).
A B
Cardiosphere-derived cells (CDCs) are immunomodulatory, limit fibrosis, and promote tissue regeneration.
Preclinical studies showed that intramyocardial administration of CDCs improved cardiac function and exercise
capacity, while increasing long-term survival in a Duchenne muscular dystrophy model. This motivated the HOPE
clinical trial with positive results after intracoronary administration of CDCs. Recently, a similar improvement was also
seen in mdx mice after systemic delivery of CDCs.
Here, we aim to evaluate multiple systemic administration of allogeneic CDCs (allo-CDCs) in promoting additional
improvement in exercise capabilities in mdx mice and to analyze the immune response after multiple administrations
of allo-CDCs.
Repeated systemic dosing of allo-CDCs resulted in increased exercise capability after two administrations in mdx
mice. CDCs showed a low immunogenic profile with weak production of alloantibodies. Steroid administration did not
impact CDC efficacy and a similar improvement in exercise capability was observed with or without steroids. In vitro,
human CDCs have a low immunogenic profile and limit proliferation of activated human T-cells.
These data demonstrate that repeat-dosing of CDCs is effective in producing additional exercise improvement in
mdx mice. Although allo-CDCs are recognized by the immune system, their low immunogenic profile and immune-
regulatory capabilities allow them to be effectively administered multiple times.
• Allogeneic CDCs given intravenously are effective in
increasing exercise capacity after first and second
administration
• Steroid administration does not impact the efficacy
of allogeneic CDCs
• CDCs have a low immunogenic profile and a strong
immunomodulation of T-cells that can limit the
inflammatory process observed in DMD patients
• These findings support clinical evaluation of
systemic administration and repeat-dosing of CAP-
1002 in patients with DMD to potentially maximize
efficacy
Figure 4. Performance of the upper limb (PUL) test was used to
measure the effects on skeletal muscle function in the HOPE I
clinical trial. A) Test description showing shoulder, middle, and
distal level tests. B) Results from middle and distal levels in
patients treated with CDCs (red) and placebo (blue).
DMD is an X-linked recessive disorder affecting
roughly 1 in 3,500 males. The disease is caused by
mutations in the dystrophin gene, causing a
truncation of dystrophin. Without dystrophin, the cell
membrane is destabilized, eventually leading to cell
apoptosis and chronic inflammation
Allogeneic CDCs Increase Exercise Capacity of mdx
Mice
D
HLA I
96%
HLA II
3.6%
Human Leukocyte Antigens (HLA)
CD803.6%
CD8657.3%
Co-Stimulatory
CD274 (PD-L1)
45.8%
CD275 (ICOS-L)
9.4%
Co-Stimulatory/Regulatory
CDCs show a low immunogenic phenotype with
expression of class I HLA, CD86 co-stimulatory
antigen, and the immuno-regulatory marker PD-L1
(Figure 7). CDCs limit proliferation of in vitro activated
human T lymphocytes.
Med
ium
PHA
CDC
5x10
9 EV/m
l
10x1
09 E
V/m
l
20x1
09 E
V/m
l 0
20
40
60
80
% p
ro
life
ra
tin
g c
ells
CD4+
CD8+
**
*
****
**
*
Inj 1 Inj 2
Figure 5. Systemic allogeneic CDCs in mdx mice. A) Exercise
capacity of CDC and vehicle-treated mice, measured weekly. B)
Serum IgG response against allogeneic CDCs after multiple
administrations.
Daily steroid administration is standard of care for
DMD patients. To measure the efficacy of CDCs in the
presence of steroids, we administered 1 mg/kg/day
prednisone to mdx mice during the week of CDC
administration. Mice underwent treadmill exercise
testing every 3 weeks to limit the learning effect seen
with weekly exercise (Figure 6).
Figure 7. Human CDC
immunology. A) Expression
of immune molecules
involved in T immune
response. B) Immune
modulation of PHA-
induced CD4+ and CD8+
T-cell proliferation by
CDCs. Results are mean
percentage values ± SEM
from 2 different donors,
each done in triplicate.
Steroid Administration Does Not Impact CDC
Efficacy
Figure 6. CDC efficacy in the presence of steroids. A) A single
injection of allo-CDCs was given at 0 weeks. Steroids were
administered daily during the week of CDC injection. Treadmill
exercise capacity was measured at weeks 0, 3, and 6. B) Immune
cells in spleen were measured for each group at the end of the
experiment. C) Serum alloantibodies against allo-CDCs were
measured at weeks 7 and 10.
Human CDC Immunology in vitro
A
B
A
B C
B
Weeks
% c
hange fro
m b
aselin
e
IMP
RO
VE
ME
NT
PHA
PBS (n=6)PBS + Steroid (n=7)Allo-CDCs (n=7)Allo-CDCs + Steroid (n=8)
PBS (n=6)Allo-CDCs (n=6)
% P
rolif
erat
ing
Cel
ls
CAP-1002 is Immunomodulatory and Regenerative
The immunomodulatory and regenerative properties
of CDCs (Figure 3) have previously been effective with
in vitro and in vivo models.
Cardiosphere-Derived Cells Are an Allogeneic Cell
Therapy Prepared from Human Heart Tissue
CAP-1002 is an allogeneic cell therapy product
based on the production of CDCs.
Figure 3. CDC mechanism of action. CDCs secrete extracellular
vesicles containing bioactive molecules that regulate different
responses involved in tissue regeneration.
Cardiosphere-Derived Cells
(CAP-1002)
Immunomodulatory
Macrophages
/T cells
Anti-apoptotic
Proliferative
Cardiomyocytes
Angiogenic
Vessels
Antifibrotic
Fibroblasts
Regenerative
Progenitor cellsAdapted from Tseliou, E. et al. J Am Coll Cardiol. 2015; 66(6):599–611.
Figure 2. CDC manufacturing scheme. A) Explant-derived cells
(EDCs) are produced from donor heart tissue and expanded to
produce a master cell bank (MCB). B) Cardiospheres are
formed from EDCs. C) CDCs are produced by culturing the cells
obtained after seeding the cardiospheres.
Donor heart MCBExplant creation
Cardiospheres CDCs Allogeneic CDC productCAP-1002
EDC outgrowth
CDC ExpansionMCB
A
B C
A