LTX-315 AND ADOPTIVE CELL THERAPY USING TUMOR-INFILTRATING LYMPHOCYTES 11567

IN PATIENTS WITH METASTATIC SOFT TISSUE SARCOMA (ATLAS-IT-04)

MORTEN NIELSEN , TINE MONBERG , BENEDETTE ALBIERI , VIBEKE SUNDVOLD , ØYSTEIN REKDAL , DORRIT HOVGAARD , MICHAEL MØRK PEDERSEN , NIELS JUNKER , INGE MARIE SVANE .
1 1 1 2 2 3 3 4 1

1
NATIONAL CENTER FOR CANCER IMMUNE THERAPY; DEPARTMENT OF 3
IDEPARTMENT OF ORTHOPEDIC SURGERY;
ONCOLOGY, COPENHAGEN UNIVERSITY HOSPITAL, HERLEV, DENMARK; 4
DEPARTMENT OF ONCOLOGY, COPENHAGEN UNIVERSITY HOSPITAL,
2
LYTIX BIOPHARMA, HERLEV, DENMARK.

Study Design (NCT03725605) Key Inclusion Criteria Characteristics of infused cells T-cell response induced against Tumor Cell Key findings of immune response data
Background • Histologically confirmed advanced/metastatic STS that is stable or has progressed TILs were successfully expanded from 4 out of 6 patient. Cultivation failed from Line (TCL)
or after minimum 1 line of systemic treatment patient 01-1004 and 01-1006, hence they were withdrawn from the study after • TILs were successfully cultivated from the LTX-315 treated tumor in
Patients with advanced stages of soft tissue sarcomas (STS) respond four out of six patients. Two patients did not receive full treatment
• At least one lesion accessible for injection Step 1.
poorly to current treatment and the prognosis is poor. Median survival (patient 01-1004, 01-1006). Common characteristics for the excluded
• At least one measurable non-injected lesion used for RECIST 1.1 response assessment patients were low abundance of TILs in the resected tumor, both at
for patients with metastatic STS at time of diagnosis was estimated 70
• ECOG Performance status (PS): 0 - 1 60 baseline and after LTX-315 treatment. Moreover, both patients had
to 10 months with a 5-year survival of 10% (1). 50 received 3 lines of prior treatment and had ECOG status 1 at screening.

no of cells (109)
Key Exclusion Criteria 40

In general, STS responds poorly to immunotherapy due to lack of 30

01-1001 01-1003 01-1007 • Exploratory analysis of immune responses were assessed for the
tumor infiltrating lymphocytes (TILs) (2,3). • A history of clinically significant active systemic autoimmune disease requiring 20
patients that received ACT of cultivated TILs (01-1001, 01-1003,
anti-inflammatory or immunosuppressive therapy within the last 3 months. 10
01-1005, 01-1007);
0
LTX-315 is a first in class non-viral oncolytic peptide that in a • Received an investigational drug therapy within 4 weeks prior to study 01-1001 01-1003 01-1005 01-1007 Heatmaps showing reactivity of PBMC and infused TIL towards autologous Tumor Cell
• External radiotherapy or cytotoxic chemotherapy within the last 4 weeks Line analysed with IFNγ ELIspot. The tumor cell line derived from tumor lesion after → Three out of four patients showed some level of reactivity against
recent Phase I/II study was shown to increase TILs in malignant Other CD3+ CD4+ CD8+
one or several Cancer Testis Antigens (CTA) in the infusion product
solid tumors after intratumoral injection (4-6). prior to study LTX-315 injections were cultivated with the indicated PBMC/TIL. Colour intensity
Bar plot shows the total number and phenotype of infused cells. shows the difference between spot count of the test samples (mean of triplicates) and in PBMC collected after EoT
• Currently taking any agent with a known effect on the immune system. Patients
Each bar shows cumulated CD4+ (green), CD8+ (blue), and other CD3+ cells (grey).
are allowed to be on a stable dose of corticosteroids (up to 10 mg daily predniso- and the negative control. Generation of TCL from pts 01-1005 failed. → Two patients (01-1003, 01-1007) showed induced reactivity against
Adoptive Cell Therapy (ACT) with TILs is a potent treatment that can #'
an autologous Tumor Cell Line (TCL) post treatment
lone or equivalent) for at least 2 weeks prior to LTX-315 administration
induce complete and durable tumor regression as documented in → One patient (01-1001) showed induced reactivity against predicted
patients with melanoma. To our knowledge, ACT has not been Treatment Schedule • Clinically active or unstable metastases in the central nervous system as assessed
by the treating physician
Anti-tumor response assessed by LTX-315 treatment induces peripheral neo-peptides
utilized for patients with advanced STS.
RECIST 1.1 expansion of T cell clones • T-cell receptor (TCR) repertoire analysis of infusion product, PBMC
Patient characteristics RECIST 1.1 and tissue samples collected from patient 01-1003 and pts 01-1007
Aim 150 Time after ACT
6 weeks
showed;
Status at ECOG PS at 12 weeks
→ LTX-315 induced expansion of a significant number of T-cell clones in
01-1001
ID Histology Sites of disease Prior systemic treatment
inclusion inclusion 6 months
This proof of concept study will evaluate the potential for LTX-315 100 01-1007

% change
Breast, lymph
Epirubicin/Cyclophosphamide, the periphery
to induce TILs prior to isolation and expansion of the TILs followed Atezolizumab, Vincristine/Ifosfamide/
01-1001 DSRCT nodes, pleura,
Doxorubicin/Etoposide, Evincristine/
PD 0
50 → Expanded peripheral T cell clones were present in tumor tissue post
by infusion of the cultured TILs to patients with advanced STS. liver
Actinomycin D/Ifosfamide UNS
01-1003
treatment
01-1002* - - - -
01-1003 Leiomyosarcoma Muscle, bone, lung Doxorubicin/Olaratumab, Olaratumab PD 0
0
01-1005 Overview of samples for immunosequencing of the TCRβ locus → New T-cell clones were detected post LTX-315 treatment, they
ACT 20 30 40
expanded significantly in the periphery, and were present in tumor

LTX-315
01-1003
01-1003 01-1007
01-1007
Doxorubicin/Cisplatin/Etoposide, Weeks from baseline ■ Comparing the TCR frequency between
Subcutis, lymph Doxorubicin/Vincristine/Actinomycin D/ T-cell clones present at baseline and tissue post treatment

PBMC post LTX-315

LTX-315
01-1004 DSRCT PD 1 post LTX-315 show evidence of clonal

PBMC post LTX-315

nodes Etoposide/Ifosfamide/

LTX-315
Cyclophosphamide, Pazopanib expansion beyond that found in healthy

LTX-315 modulates the tumor environment controls over a two-week period

post
01-1005 Solitary fibrous tumour Abdomen Doxorubicin/Dexrazoxane SD 1 (~10 clones).

PBMCpost
17 17

Objectives Vincristine/Ifosfamide/Doxorubicin/
LTX-315
LTX-315
(Step 1)modulates
modulatesthe
thetumor
tumorenvironment
environment ■ Both patients show evidence of

PBMC
Etoposide newly detected T-cell clones post
Sclerosing epithelial LTX-315 treatment (along the y-axis)
01-1006 Eye, bone Vincristine/Actinomycin D/Ifosfamide PD 1

Conclusion
fibrosarcoma PBMC baseline
baseline PBMC baseline

LTX-315`s unique mode of action

PBMC PBMC baseline
Gemcitabin/Docetaxel,
Primary Gemcitabin/Paclitaxel, Gemcitabine IHCIHC
CD4IHC
CD4CD4 IHCIHC
IHC CD8
CD8CD8

• Ability of LTX-315 to induce T-cell infiltration prior to TIL expansion in 1200 1200 800 800

Whole tumour

Whole tumour
results in immunogenic cell death

Whole tumour
tumour

Whole tumour
tumour
Parotid gland, 1000 1000
01-1007 Solitary fibrous tumour Doxorubicin/Dexrazoxane PD 0 600 600
advanced STS
Treatment-associated peripheral expansion
lung, liver, kidney
• In this hard-to-treat patient population, LTX-315 in combination
800 800

cells/mm2

cells/mm2

cells/mm2

cells/mm2
600 600 400 400

resulting in effective release of • Safety of LTX-315 as part of adoptive T-cell therapy in advanced STS with ACT therapy was able to stabilize the disease in 3 out of 4
400 400

Whole

Whole
200 200

is maintained 6 months post treatment

200 200

patients that received the full treatment.

0 0 *NA *NA 0 0 *NA *NA

potent immunostimulants and Secondary

Adverse Events during therapy
1200 1200
1000 1000
800 800

• LTX-315 was well tolerated with adverse events being

Invasive margin
margin

Invasive margin
margin
600 600

Invasive margin

Invasive margin
• Ability to expand CD8+ T-cells from tumor tissues
800 800

cells/mm2

cells/mm2

cells/mm2

cells/mm2
antigens followed by a broad T-cell
600 600 400 400
Peripheral expansion of T-cell clones from baseline ■ Both LTX-315 and ACT result in
mostly mild or moderate in severity.
Adverse events reported by the investigator as related to LTX-315 and Adoptive 400 400
peripheral expansion
• Anti-tumor activity of LTX-315 as a part of ATC therapy in advanced STS

Invasive

Invasive
200 200
200 200
Cell Transfer therapy
response (4-11)
0 NA NANA NA NA *NANA
NA*NA NANA NA
NA NA
• The treatment was shown to generate tumor-specific T cells,
0 0 NA
0 NANA NA NA *NA
NANA*NA NANA NA
NA NA
■ High levels of clonal expansion are

bystander…

bystander…

bystander…

bystander…
post LTX-315

post LTX-315

post LTX-315

post LTX-315
post LTX-315

post LTX-315
post LTX-315

post LTX-315
post LTX-315

post LTX-315

post LTX-315

post LTX-315

post LTX-315

post LTX-315
post LTX-315

post LTX-315
post LTX-315

post LTX-315
post LTX-315

post LTX-315
post LTX-315

post LTX-315

post LTX-315

post LTX-315
baseline

baseline
baseline

baseline

baseline

baseline
baseline

baseline
baseline

baseline
baseline

baseline
baseline

baseline

baseline

baseline
baseline

baseline
baseline

baseline
Baseline

Baseline

Baseline

Baseline
observed 6 months post ACT
Exploratory expand T-cell clones in the periphery, and generate de novo
Therapy AE Grade 1-2 (n) Grade 3-4 (n) ■ Peripheral expansion post LTX-315
• Assess tumor antigen specificity LTX-315 Diarrhoea 1 0
01-100101-1001
01-100301-1003
01-100401-1004
01-100501-1005
01-100601-1006
01-100701-1007 01-100101-1001
01-100301-1003
01-100401-1004
01-100501-1005
01-100601-1006
01-100701-1007
was more pronounced than post ACT T-cell clones.
*H&E*H&E
staining
staining
of theoftumor
the tumor
from pts
from01-1005
pts 01-1005
showed
showed
completely
completely
necrosis
necrosis
post treatment
post treatment
and IHC
andwas
IHCtherefore
was therefore
not assessed
not assessed in patient 01-1003
Fever 1 0
• Investigate and characterize immune status and nature of anti-tumor immune Itching
Pain at injection site
1
4
0
0
*H&E staining of the tumor from pts 01-1005 showed complete necrosis post treatment
and IHC was therefore not assessed
■ Patient 01-1007 showed very high
levels of expansion after ACT therapy
responses Redness at injections site 1 0
Stomach pain 1 0
Chemotherapy Anaemia
Constipation
Diarrhoea
3
1
1
0
0
0
Modulation of the systemic immune
REFERENCES
Dry mouth 1 0
response
Endpoints
Fatigue 1 1

T-cell clones that expand post baseline

Headache 1 0
Haematuria 1 0 1. Sorbye, S.W. et al.; PlosOne (2011) 6. Spicer, J. et al.; Cancer Research (2021)
Hot flashes 1 0
T-cell response induced against tumor antigens 2. Smolle, M.A. et al.; Oncoimmunology (2021) 7. Jebsen,N.L. et al.; Journal of Medical Case

are present in injected lesion and in the

Hypocalcaemia 1 0
3. Sousa, L.M. et al.; Cancers (2021). reports (2019)
Primary Hypomagnesaemia 1 0
4. Haug, B.E. et al.; Journal of Medicinal Chemistry 8. Liao H-W. et al.; Cell Stress (2019)
Hyponatraemia 1 0

• Change in T-cell level in tumour tissue from Baseline to end of Step 1

bystander lesion
Hypophosphatemia 1 0 200 (2016) 9. Nestvold,J. et al.; Oncoimmunology (2017)
Nausea 4 0
5. Sveinbjørnsson, B. et al.; Future Medicinal 10. Eike, L-M. et al.; Oncotarget (2015)
Neutropenia 0 4
• Adverse Events (AE) related to LTX-315 or the combination of LTX-315 and Rash 2 0 Chemistry (2017) 11. Forveille, S. et al.; Cell Cycle (2015)
Stomach-ache 1 0
ACT from Baseline to end of treament (EoT) (Step 2) Stomatitis 1 0 100

■ Many of the expanded ■ Patient 01-1003; tissue

Thrombocytopenia 0 4
Vomiting 1 0 T-cell clones in patient biopsy post LTX-315 was
Secondary TIL Chills 2 0 01-1007 (grey bar) were not available and similar
Diarrhoea 1 0 also present in the injected analysis is therefore not Collaborators
• Total number of CD8+ T cells in the final infusion product Dyspnoea
Fatigue
0
0
1
1
0
lesion (blue bar) and in the
bystander lesion (green
included.
Fever 2 2
• Anti-tumor effect assed by ORR, CBR and PFS Headache 1 0
bar) post treatment.
Rash 1 0
IL2 Anaemia 2 0 01-1001 01-1003 01-1005 01-1007 ■ The number of expanded
Exploratory Diarrhoea 3 0 T-cell clones that were
Elevated alkaline phosphate 1 0 present in both injected
Heatmaps showing reactivity of PBMC and infused TIL towards predicted neo-peptides
• Identification of tumor-antigen specific T cells in tumor tissue and peripheral blood Elevated liver enzymes 1 0 lesion and bystander
Fever 2 2 and selected Cancer Testis Antigens analysed with IFNγ ELIspot. Colour intensity shows Injected lesion baseline (T-clones <100)
increased over time.
• Change in immunological parameters from Baseline to 15 m after EoT.
Headache 1 0
Rash 1 0 the difference between spot count of the test samples (mean of triplicates) and the Injected lesion 5 weeks post LTX-315
Bystander lesion 12 weeks post ACT
Vomiting 1 0
negative control.

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