GENETIC REPROGRAMMING OF TAMS BY ENGINEERED EXOSOMES RESULTS IN POTENT SINGLE AGENT ANTI-TUMOR ACTIVITY Sushrut Kamerkar1, Dalia Burzyn1, Charan Leng1, Olga Burenkova1, Su Chul Jang1, Raymond Yang1, Katherin Kirwin1, Tong Zi1, William Dahlberg1, Eric Zhang1, Scott Estes1, Sylvie Maubant2, Olivier Duchamp2, Kyriakos Economides1, Timothy Soos1, Sriram Sathyanarayanan1.

1. Codiak BioSciences, Cambridge, MA.2. Oncodesign, Dijon, France

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AACR ANNUAL MEETINGJUNE 23, 2020

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Disclosure Information

AACR Annual Meeting 2020

Dalia Burzyn

• I have the following financial relationships to disclose:- Employee of Codiak BioSciences

• I will not discuss off label use and/or investigational use in my presentation

3

M2-like tumor-associated macrophages (TAMs) promote cancer progression

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Jackute et al. BMC Immunology 19:3, 2018

Total M2 macrophage density correlation to overall survival in lung cancer

Anfray et al. Cells, 9, 46, 2020

Reprogramming TAMs by targeting key “undruggable” pathways

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exoASO-STAT6 or C/EBPb

Ø Selective modulation of key M2 transcription factors by combining:

Ø Preferential tropism of exosomes to myeloid cells

Ø Codiak’s exoASO™ platform to load exosomes with specific antisense oligonucleotides (ASOs)

exosomeSTAT6 or C/EBPb antisense oligonucleotide

TAM reprogramming

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0 6 12 18 24 30 36 42 480

5×107

1×108

1.5×108

Time (Hours)

Tota

l Obj

ect I

nten

sity

(RC

U x

um/im

age)

Exo-ASO-M2

Free-ASO-M2

Exo-ASO M0

Free ASO-M0

Exosome-mediated selective uptake in human M2 macrophages

exoASO shows enhanced tropism to M2 macrophages in vitro

Ø Exosome surface protein PTGRFN may mediate selective delivery to macrophages and other myeloid cells via the cell type specific expression of its cognate receptors in these cells

PTGFRN cognate receptors in human GBMM2 mac + exoASO

M2 mac + Free ASO

In vivo exosome-mediated selective delivery of ASOs to macrophages and MDSCs

6

LIVER (IV) BLOOD (IV)

CD45+

CD45-

Kupffer

cells

CD11b+

F4/80-

CD11b+

Ly6C+

CD11b+

Ly6G+

0

500000

1000000

1500000

2000000

Cy5

MFI

(m

inus

PB

S co

ntro

l)

Exo ASOFree ASO

9x

B cell

T cell

NK cell

Macs

Red pulp

macs

Monocytes

mMDSC

gMDSCs

/neutr

0

10000

20000

30000

40000

50000

Cy5

MFI

(m

inus

PB

S co

ntro

l)

Exo ASOFree ASO 3.4x 2.2x 2.7x

B cells

T cells

NK cells

CD11b+ D

Cs

CD11b- D

Cs

Monocytes

mMDSCs

/othergMDSC

/neutro

phil

0

50000

100000

150000

200000

Cy5

MFI

(m

inus

PB

S co

ntro

l)

Exo ASOFree ASO 12x

11x

11x

TUMOR (IT)

Nor

mal

ized

coun

ts

ASO-Cy5

— Unlabeled exo— Macrophages_exoASO— Tumor cells_exoASO

SPLEEN (IV)

2x

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0.0

0.2

0.4

0.6

0.8

1.0

1.2

102 103 104

CEBPB

concentration (nM)Nor

mal

ized

% g

ene

expr

essi

on (

hCEB

PB)

0

C/EBPb exoASOC/EBPb free ASOUntreated

Scramble exoASO

STAT6 and C/EBPb silencing by exoASO repolarizes primary human M2 macrophages in vitro

7

0.0

0.2

0.4

0.6

0.8

1.0

1.2

102 103 104

STAT6

concentration (nM)Norm

aliz

ed %

gen

e ex

pres

sion

(hS

TAT6

)

0

STAT6 exoASOSTAT6 free ASOUntreatedScramble exoASO

IC50 (nM)

STAT6 exoASO 185.8

STAT6 free ASO 366.4 Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

200

400

600

800

CD163

mRN

A no

rmal

ized

line

ar c

ount

s

********

Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

500

1000

1500

TGFb1

mRN

A no

rmal

ized

line

ar c

ount

s

********

Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

100

200

300

CD206

mRN

A no

rmal

ized

line

ar c

ount

s

********

Target gene knockdown

Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

200

400

600

800

TNFa

pg/m

lLLOQ

****

Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

1000

2000

3000

4000

IL12p40

pg/m

l

LLOQ

******

Effective macrophage reprogramming

IC50 (nM)

C/EBPβ exoASO 427.1

C/EBPβ free ASO 982.0 Untreate

d

Scramble

exoASO

STAT6 exo

ASO

C/EBPb ex

oASO0

500

1000

1500

2000

2500

TARC/CCL17

pg/m

l

*****

Codiak Proprietary Information – Not for Further Dissemination or Use

exoASO-STAT6 and C/EBPβ induce in vivo knockdown and reprograming of TAMs

8

Target gene knockdown

Reprograming to anti-tumoral phenotype

D0

CT26, SC

D8

IT Injection

D10 D13 D14 After intratumoral treatment, tumor-associated myeloid cells were isolated and gene expression was analyzed

STAT6 Free ASO

Scramble ExoASO

C/EBPbExoASO

STAT6 ExoASO

Scramble ExoASO

STAT6ExoASO

STAT6 FreeASO

Scramble Exo ASO

Stat 6

Free ASO

Stat 6

Exo ASO0.00

0.25

0.50

0.75

1.00

1.25

1.50

Norm

alize

d Ge

ne E

xpre

ssio

n (m

Stat

6) CD11b Enriched

***

**18%

49%

0.00

0.25

0.50

0.75

1.00

1.25

1.50

Norm

alize

d Ge

ne E

xpre

ssio

n (m

Cebp

B)

**

CD11b Enriched

Scramble ExoASO

CEBPbExoASO

Scramble

exoASO

STAT6 free

ASO

STAT6 exo

ASO

C/EBPb ex

oASO0

500

1000

1500

2000

2500

CSF1R

mRN

A no

rmal

ized

line

ar c

ount

s

********

Scramble

exoASO

STAT6 free

ASO

STAT6 exo

ASO

C/EBPb ex

oASO0

50

100

150

200

250

NOS2

mRN

A no

rmal

ized

line

ar c

ount

s

****

45%

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0 20 40 600

50

100

Days Post Tumor Implatation

Perc

ent s

urvi

val

exoASO-Scramble (35)PBS + Iso (30)Free ASO C/EBPb (30)exoASO-C/EBPb (50)

anti-PD1 (35)exoASO + anti-PD1 (Undef.)Free ASO + anti-PD1 (39.5)anti-CSF1R (30)

Potent monotherapy and combination antitumor activity with exoASO-C/EBPb and STAT6

9

Antitumoral efficacy Survival

* Similar anti-tumor activity with STAT6

0250500750

100012501500175020002250

exoASO Scramble PBS Free C/EBPb ASO

0250500750

100012501500175020002250

exoASO C/EBPb anti-PD1

Free C/EBPb ASO+PD1

exoASO C/EBPb+PD1 anti-CSF1R

6/10 CR 8/10 CR 0/10 CR

exoASO C/EBPβ+aPD1à Undef.

exoASO C/EBPβà 50 days

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Systemic administration of exoASO-C/EBPb results in potent anti-tumor response in an HCC model

10

D0

InoculationIV injection, TIWx2, 6 total

D3 D14 D16IP injection, DIWx2, 4 total

Randomization

End of Study

Vehicle

CebpB exoASO CebpB+aPD1

Scramble+aPD1

Vehicl

e

exoASO

Scramble+

PD1ex

oASO

C/EBPbex

oASO

C/EBPb+P

D1

Free C/EBPb

ASO0.0

0.5

1.0

1.5

2.0

Target gene knockdown: C/EBPb

Nor

mal

ized

Gen

e E

xpre

ssio

n (m

Ceb

pB)

***

Vehicl

e

anti P

D1

exoASO

Scramble+

aPD1

exoASO

C/EBPbex

oASO

C/EBPb+a

PD1

Free C/EBPb

ASO

anti C

SF1R0

50

100

150

% Scored lesions

% S

core

d le

sion

s (t

umor

inva

sion

)

*** *

**

CR:7/15

CR:5/7

CR:1/8

CR:0/8

Vehicle exoASO-C/EBPb

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11

STAT6 and C/EBPβ are highly expressed in human tumor macrophages and correlate with poor prognosis

Expression in human GBM Correlation with survival in human HCC

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12

exoASO: a first-in-class strategy to target TAMs in a highly selective manner

exoASO-STAT6 or C/EBPb• Developed a novel therapeutic to target

and reprogram pathogenic M2 macrophages in cancer

ANTI-TUMORAL EFFICACY

• Treatment with exoASOs against STAT6 and C/EBPβ results in potent anti-tumor efficacy via local and systemic administration

DELIVERY TO TAMs

• Selective targeting of M2 macrophages in the tumor microenvironment (TME)

TARGETING STAT6 and C/EBPβ• Targeting STAT6 and C/EBPβ induces

higher target gene knockdown and superior macrophage reprogramming vs Free ASO, both in vitro and in vivo

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