Image-guided cancer gene therapy in combination with advanced chemotherapeutics
Project Title: Image-guided cancer gene therapy in combination with advanced chemotherapeutics
Funding: EEA and Norwegian Financial Mechanisms 2009-2014 Programme Programme LV05 “Research and Scholarships”.
Project No.: NFI/R/2014/051
Period: 24 month (1st May 2015–30st April 2017)
Partners: Oslo University Hospital (OUH); Latvian Institute of Organic Synthesis (LIOSI)
Project costs: 478 440,00 EUR (BMC – 150 000,00 EUR; OUH – 188 440,00 EUR; LIOSI – 140 00,00 EUR)
Principle investigator: Dr.biol. Anna Zajakina
Project Investigator, coordinator Dr.biol. Anna Zajakina (BMC) - firstname.lastname@example.org
WP leader and Member of Steering Committee Alexandre Corthay, PhDy (OUH) - email@example.com
WP leader and Member of Steering Committee Dr.habil.chem Gunars Duburs (LOSI) - firstname.lastname@example.org
Project Manager (BMC) – email@example.com
The project presents a new collaborative research initiative between Oslo University Hospital, Latvian Biomedical Research and Study Centre and Latvian Institute of Organic Synthesis. Our main objective is to perform preclinical evaluation of a novel therapeutic strategy against cancer cells based on combination of chemotherapeutic drugs with viral gene therapy vectors. We propose to apply novel alphaviral vectors encoding cytokines, which able to stimulate tumor-associated macrophage (TAM) infiltration into the tumor and induction of pro-inflammatory microenvironment to block tumor recovery after chemotherapy. The outcomes of treatment will be monitored using advanced imaging technologies including bioluminescent and fluorescent imaging. The project will contribute to research excellence both in Latvia and Norway and will stimulate the development of innovative therapies urgently needed to improve the standard therapeutic approaches for cancer.
- 12.05.2015 project kick-off teleconference
- 05.06.2015 project seminar at LIOS
- Development of liposomal nanoparticles for delivery of therapeutic genes and chemotherapeutic drugs is started
- Construction and synthesis of alphaviral DNA vectors is started
- Development of human PDX tumor models is started
- First liposomal compounds and fluorescent dyes were synthesised
- New alphaviral vectors producing anti-cancer citokines were constructed and synthesized
- Human PDX tumor model in immunodeficient mice was developed.
November (2015) - January (2016)
- 17.11.2015 progress report seminar at LBMC
- First data obtained for alphaviral vector ability to express anti-tumor cytokines in cancer cells
- New fluorescent dyes were tested for the ability to stain cancer cell cultures
- Macrophage polarization experiments have started
Within the first reporting period, the involved three groups of scientists from Latvian Biomedical Research and Study Centre (LBMC), Latvian Institute of Organic Synthesis (LIOS) and Oslo University Hospital (OUH) have finished the project activities devoted to establishment of tumor models, synthesis of drug candidates and potent cancer gene therapy vectors. The Tumor Immunology group headed by Dr. Corthay (OUH) has finalized the first detailed characterization of the “Immune Cell Composition in Human Non-small Cell Lung Cancer” (a manuscript is submitted for publication). This knowledge will serve as a basis for the planned immunotherapy experiments. The group has also established the patient-derived xenotransplantation (PDX) mouse model for lung cancer that will be used to test immunotherapy protocols with alphaviruses.
At the same time, the Latvian Cancer Gene Therapy group headed by Dr. Zajakina (LBMC) has produced first alphaviral vectors encoding therapeutic gene candidates for reprogramming of tumor microenvironment to neglect the immune tolerance to cancer cells in breast cancer and lung cancer models. Novel constructs for expression of mouse and human interferon gamma and tumor necrosis factor alpha will be used in combination with advanced chemotherapeutics synthesized by the group of Prof. Duburs (LIOS). Series of putative delivery agents on the basis of 1,4-dihydropyridine (1,4-DHP) core were obtained by a variation of different alkyl moieties at the 3,4 and 5 positions of 1,4-DHP cycle (a manuscript is submitted for publication). Moreover, synthesis and characterization of fluorescent compounds comprising electron donor groups and conjugated electron accepting moieties was performed in order to obtain polychromic probes possessing near infrared (NIR) emission for efficient fluorescent imaging and drug tracking in vivo.
Furthermore, LBMC group together with OUH colleagues have finished experiments related to proteome analysis of B16 mouse melanoma cells susceptible and unsusceptible to alphavirus infection. These results pave the way for optimized development of strategies for designing tumor tropism of alphaviral vectors as potent cancer gene therapy tool. All together the first project period has shown the successful implementation of the proposed research ideas and fruitful results of the complementary collaboration between the partners.
February - April 2016
It is known that interferon-γ (IFN-γ) primes macrophages for enhanced inflammatory activation and acquisition of M1 phenotype by Toll-like receptors (TLRs). As M1 macrophages have anti-tumorigenic and tumoricidal functions they could be used in cancer immunotherapy. Whereas tumour necrosis factor-a (TNFa) has been suggested to have cancer-suppressive properties as well. Though TNFa is a potential therapeutic for cancer immunotherapy, targeted delivery of TNFa to tumours is needed to limit its toxicity.
In order to investigate the potential of cytokine-expressing Semliki Forest virus (SFV) based vectors in cancer immunotherapy, three new SFV vector based alphaviral constructs incorporating mTNFa, mIFNg and hIFNg genes were developed. SFV based vectors were shown to infect Lewis Lung carcinoma (LLC) cells and greatly inhibit their growth in vitro. In order to study whether SFV-mIFNg derived IFNg could enhance macrophage activation through TLRs in vitro, bone marrow derived macrophages (BMDM) were challenged with SFV-mIFNg derived mIFNg and effect on LLC cell growth was detected. SFV derived mIFNg functionality in vitro was shown in two independent experiments. It was concluded that SFV derived mIFNg could be used for enhanced activation of macrophages to cancer suppressive phenotype through different TLRs.
Future in vivo experiments comprises the use of SFV-mTNFa for cancer growth inhibition in combination with SFV-mIFNg whereas SFV-hIFNg construct will be used on human lung cancer xenotransplantant model in vivo.
May - July 2016
The involved three groups of scientists from Latvian Biomedical Research and Study Centre (LBMC), Latvian Institute of Organic Synthesis (LIOS) and Oslo University Hospital (OUH) continued the project implementation activities according to the work plan. The synthesised drug candidates (LIOS) and viral vectors (LBMC) were tested in pilot experiments using in vitro and in vivo cancer models established at OUH. The primary results indicate that Semliki Forest virus (SFV) vectors expressing macrophage stimulating cytokines (produced at LBMC) can activate tumor associated macrophages to kill cancer cells in presence of specific TLR ligands. The treatment with SFV vectors alone or together with TLR ligands was investigated in tumor growth inhibition experiments (OUH). The pilot results have demonstrated positive therapeutic outcome of the proposed treatment strategy.
Furthermore, the LIOS synthesised fluorescent compounds showed a high potential to be used as cancer cell tracking agents in vivo in mice models. These novel fluorescent compounds possessing fluorescent properties within the infrared excitation spectrum can be further developed as theranostic drugs for cancer diagnostic and therapy.
In frame of the project, three international pier reviewed articles were published:
1. Joly-Battaglini A, Hammarström C, Stankovic B, Aamodt H, Stjärne J, Brustugun OT, Helland Å, Øynebråten I, Corthay A. Rituximab efficiently depletes B cells in lung tumors and normal lung tissue. F1000Res. 2016 Jan 8;5:38. doi: 10.12688/f1000research.7599.1.
2. Rufus Smits, Baiba Turovska, Sergey Belyakov, Aiva Plotniece, Gunars Duburs Synthesis of 5-carboxy-6-methyl-3,4-dihydro-2(1H)-pyridone derivatives and their electrochemical oxidation to 2-pyridones. Chemical Physics Letters. Volume 649, April 2016, Pages 84–87.
3. Jelena Vasilevska, Gustavo Antonio De Souza, Maria Stensland, Dace Skrastina, Dmitry Zhulenvovs, Raimonds Paplausks, Baiba Kurena, Tatjana Kozlovska, Anna Zajakina. Comparative protein profiling of B16 mouse melanoma cells susceptible and non-susceptible to alphavirus infection: effect of the tumor microenvironment. Cancer Biology & Therapy. Accepted for publication.
More information about the EEA and Norway grants can be found at the State Education Development Agency (VIAA in Latvian) web page
and at the EEA and Norway Grants web page
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