Facilitating T-cell therapy in solid

tumors with oncolytic adenovirus

Joao Santos1,2, Mikko Siurala1,2, Riikka Havunen1,2, Victor Cervera-Carrascon1,2,

Suvi Sorsa1, Akseli Hemminki1,2,3

1TILT Biotherapeutics Ltd, Helsinki, Finland;´2Cancer Gene Therapy Group, Department of Oncology, Faculty of

Medicine, University of Helsinki, Helsinki, Finland; 3Helsinki University Hospital Comprehensive Cancer Center,

Helsinki, Finland.

Background

Conclusions

Results1. Adenovirus coding for TNFa and IL-2

improves the B16.OVA growth control in

combination with OT-I cell therapy1

• Tumor-infiltrating lymphocyte (TIL) or gene-engineered T-cell

therapy have efficacy limitations such as suboptimal trafficking

of transferred cells to tumors

• Lymphodepleting preconditioning and high-dose IL-2

postconditioning are major sources of toxicity in adoptive

T-cell therapy protocols

• Programmed Death (PD)-1 blockade therapy demonstrates

good clinical results in only in TIL infiltrated tumors

• TILT-123 (Ad5/3-E2F-D24-hTNFa-IRES-hIL2) is an oncolytic

virus designed to enable T-cell therapy coding for:

• Human Tumor Necrosis Factor alpha (TNFa)

– Increases T-cell trafficking

• Human interleukin-2 (IL-2) – Increases T-cell

survival and growth

2. TILT-123 enables complete tumor

regression and protection from tumor

rechallenge in TIL treated hamsters2

6. Prime & boost adenovirus therapy

regimen enables 100% survival in

anti-PD-1 treated mice6

5. TILT-123 improves the survival and

efficacy of animals treated with adoptive

T-cell transfer without preconditioning5

4. Adenovirus-mediated local IL-2

delivery is superior to postconditioning

with systemic high-dose IL-24

3. TILT-123 leads to regression and

transduction of distant tumors in a

TIL therapy setting3

(A) HapT1 tumor-bearing hamsters were treated 5 times with 1 × 109 VPs of TILT-123 and once with

4×106 TILs both injected i.t. (n = 6–7). (B) Complete tumor remission was evaluated before animals

were euthanized. (C) Hamsters previously cured of HapT1 tumors were re-challenged with the same

HapT1 tumor cells or (D) with a distinct cell line (DDT1-MF2).

(A) Combined adenoviruses coding for mTNFa and mIL2 (1:1 ratio) were used to treat B16.OVA tumors in

combination with adoptive transfer of 1.5×106 CD8-enriched OT-I cells. (B) B16.OVA tumor-bearing mice were

administered 6 × 106 111Indium-oxine labeled OT-I cells intraperitoneally (i.p.) with simultaneous intratumoral

(i.t.)injection of cytokine-coding adenoviruses. Mice were imaged on 24, 48, and 96 hours after treatment

through SPECT/CT. (C) Representative SPECT/CT images of 96 hours timepoint with white circles indicating

the locations of subcutaneous tumors.

Hamsters bearing 2 HapT1 tumors were infused with 5x107 TILs

received 5 injections of 1x108 VPs of TILT-123 i.t. (A) The growth of

injected and (B) non-injected hamster tumors (n=5-6) was measured

every two to three days until day 33. (C) Small amounts of viral DNA

was detectable also in non-injected tumors on day 16. (D) No

differences were observed between the injected and non-injected

tumor sizes on day 33.

(A) 1 × 106 OT-I cells were infused in B16.OVA tumor-bearing mice that were treated 5 + 5 days during a period

of 12 days with mIL‐2 (100,000 IU) i.p. or once a week with 1 × 108 vp of Ad5‐CMV‐mIL‐2 i.t. (n=7-9). (B)

Levels of mIL‐2 were measured from end‐point tumors and serums with cytometric bead array.

(A) HapT1 tumor-bearing hamsters received 5x107 TILs i.p. and 4 rounds of 5 injections of 1x108 VPs of

TILT-123 intratumorally and/or lymphodepleting preconditioning. Survival was followed during 121 days

(n=4-5). (B) Mice tumor growth followed during 11 days (n=7-10).

(A) Combined adenoviruses coding for mTNFa and mIL2 (1:1 ratio) were used to treat B16.OVA tumors in

combination with i.p. administration of 0.1 mg of anti-PD-1 (repeated every 3 days for a total of 13–15 times)

(n=12-16). (B) Overall survival. (C) Individual tumor growth lines for the different conditions tested.

References

TILT-123 viral particles (VPs) transduce and lyse tumor cells releasing TNFa, IL-2 and danger signals in the

tumor microenvironment. This will activate and attract TILs to the tumor site that were either adoptively

transferred or potentiated through inhibition of the PD-1 pathway. Consequently, the efficacy of adoptive T-cell

or anti-PD-1 therapy will be enhanced.

D.C.

A. B. C.

A. B.

A.

A. B.

A. B.

C.

TILT Biotherapeutics Ltd

Haartmaninkatu 3, 4th floor, C-wing

00290 Helsinki, Finland

For more information:

joao@tiltbio.com

• Adenovirus-mediated delivery of TNFa and IL-2

enables adoptive T-cell therapy using TIL therapy or

gene-modified T-cells

• TILT-123 induces systemic antitumor responses and

tumor-specific memory in animals treated with TIL

therapy

• TILT-123 replaces high-dose IL-2 postconditioning

and lymphodepleting preconditioning in a setting of

adoptive T-cell therapy

• Adenovirus therapy enables complete responses in

animals receving anti-PD-1 therapy

• Clinical translation is underway to validate these

hypotheses

™

A. B.

1Siurala M. et al; Mol Ther (2016)2Havunen R. et al; Mol Ther Onc (2017)3Havunen R. et al; Submitted4Santos JM. et al; Int J Cancer (2017)5Santos JM. et al; Submitted6Cervera-Carrascon V. et al; Oncoimmunology (2017)

D.

B.

C.