Local immunotherapy with ONCOS-102 shapes harmful tumor associated CD68+ macrophages to become beneficial cells that correlate with increased overall survival

Pesonen S1, Lundin J2, Linder N2, Turkki R2, Ristimäki A3, Joensuu T4, Kairemo K4, Partanen K4, Alanko T4, Jäger E5, Karbach J5, Wahle C5, Hemminki A6, Backman C1, von Euler M1, Hakonen T1, Ranki T1, Vuolanto A1, Jaderberg M1. 1Targovax Oy, Helsinki, Finland, 2Institute for Molecular Medicine Finland (FIMM), Helsinki, Finland, 3Division of Pathology, HUSLAB and Haartman Institute, Helsinki University Central Hospital, Helsinki, Finland, 4Docrates Cancer Center, Helsinki, Finland, 5Hämatologie-Onkologie, Krankenhaus Nordwest, Frankfurt, Germany, 6Cancer Gene Therapy Group, Haartman Institute, University of Helsinki, Helsinki, Finland.

ONCOS-102 (Ad5/3-D24-GMCSF) is a tumor-targeted oncolyticadenovirus coding for human GM-CSF

Intratumoral ONCOS-102 induces a systemic CD8+ T cell response against  patient’s  unique  cancer  cells:

INTRODUCTION

Innate Immune System Adaptive Immune System Anti-tumor Immune Response

”Recognition  of  threat” ”T  cell  activation” ”Immune  attack”

tumor lymph node tumor

metastasisStrongco-stimulation

ONCOS-102

T cellactivation

SystemicT cell attack

Multiple activatory mechanisms:• Immunogenic cell death• Pro-inflammatory cytokines• Release of tumor antigens• Local GMCSF expression

Targeted anti-tumor immune response:• ONCOS-102 teaches immune

system to recognize unique cancer cells of each patient

Systemic anti-tumor immune response• In situ vaccination• Immunological memory

provides long term protection

• High density of tumor infiltrating CD68+ macrophages at baseline was associated with short survival suggesting that these macrophages had tumor promoting phenotype

• ONCOS-102 –triggered CD68+ cell infiltration correlated with prolonged survival suggesting that these cells had different phenotype from CD68+ cells present in tumors before ONCOS-102 treatment

• Infiltration of multiple innate and adaptive immune cell populations after ONCOS-102 administration correlated with increased OS

• ONCOS-102 has potential to activate immunologically silent tumor and reduce the local immune suppression in advanced tumors

Phase I study - design 12 last-line patients with 100% chemo refractory solid tumors were treated at 3 dose levels (3+3+6 pts)

0 1 4 8 15 29 57 113 141 169Day

ONCOS-102

85

Biopsy

PET / CT

X X X X X X X X XX X X

X X X

PBMCs X X X X X X X X X

Dose cohorts: 3x1010, 1x1011, 3x1011 viral particles

01 02 04 06 08 09 13 14 15 17 18 19

CD68

(Abs

olute

expr

essio

n)

Patient number

0.000

0.020

0.040

0.060

0.080

0.1000.150

Heterogeneity in CD68+ macrophage density was seen in tumors before and after ONCOS-102 administration

Pt 06

Pt 02

Pt 19

Pt 08

Baseline After

CD68+ cells in tumors (IHC)

Figure 3. The density of CD68+macrophages in tumor biopsies wasassessed by quantitativeimmunohistochemistry. A correlationbetween absolute expression levelof CD68+ macrophages in tumorsand overall survival (OS) wasassessed by Spearman´s rankcorrelation analysis.

Infiltration of innate and adaptive immune cells into tumors was seen following ONCOS-102 administration

1,E-02

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

1,E+04

1,E-02

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

1,E+04

1,E-02

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

1,E+04 CD68+ macrophages CD8+ T cells CD4+ T cells

Immu

ne ce

lls in

tumo

rs(F

old ch

ange

from

base

line)

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19** *

** *

01 02 04 06 08 09 13 14 15 17 18 19

SD at 3 months (RECIST)

PD at 3 months (RECIST)

Post-treatment induction of tumor-specific CD8+ T cells*

CD68+ cells in baseline tumors were associated with short OS while post-treatment increase in CD68+ cells correlated with increased OS

Post-treatment increase in tumor infiltrating innate and adaptive immune cells was associated with increased OS

CD68

+ ce

lls in

tumo

rs(a

bsolu

te ex

pres

sion)

Survival (months)

Tumor afterONCOS-102

Tumor beforeONCOS-102

Baseline After ONCOS-102

Survival (months)

1,E-04

1,E-03

1,E-02

1,E-01

0 10 20 301,E-03

1,E-02

1,E-01

1,E+00

0 10 20 30

(alive)

(alive)

r= - 0.59, p= 0.04 r= 0.71, p= 0.01

Figure 2. Immune cells in tumors were assessed by IHC before and after ONCOS-102 administration.

CD68

+ ce

lls in

tumo

rs(a

bsolu

te ex

pres

sion)

1,E-02

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

0 5 10 15 20 25

CD11c+(Dendritic cells)

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

1,E+04

0 5 10 15 20 25

CD8+(Cytotoxic T cells)

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

0 5 10 15 20 25

CD3+(All T cells)

1,E-01

1,E+00

1,E+01

1,E+02

1,E+03

0 5 10 15 20 25

CD163+(Macrophages)

Immu

ne ce

lls in

tumo

rs(F

old-ch

ange

from

base

line)

Survival (months) Survival (months) Survival (months)Figure 4. Frequency of immune cells in tumor biopsies were measured by immunohistochemistry before and afterONCOS-102 administration. A correlation between post-treatment increase in immune cells and overall survival (OS)was assessed by Spearman´s rank correlation analysis.

(alive)(alive) (alive)

(alive)

CONCLUSIONS

Figure 1. Sequential biopsies were collected at baseline and after ONCOS-102 treatment initiation and analyzed for the presence of immune cells by immunohistochemistry (IHC) in digitally scanned samples. Readout of the expression levels for immune cells was performed by the use of an image analysis algorithm based on color deconvolution and segmentation of the IHC stained cells.

RECIST evaluation not done

Survival (months)