Download - Allogeneic CDCs have a low immunogenic profile and repeat … · 2018-04-19 · human CDCs have a low immunogenic profile and limit proliferation of activated human T-cells. These

Transcript
Page 1: Allogeneic CDCs have a low immunogenic profile and repeat … · 2018-04-19 · human CDCs have a low immunogenic profile and limit proliferation of activated human T-cells. These

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