A Novel approach for cancer treatment by activated monocyte - derived macrophages

Project leaders

Francesca Mariani, Andrey Tchorbanov

Agreement

BULGARIA - BAS - Bulgarian Academy of Sciences

Call

CNR-BAS 2016-2018

Department

Biomedical sciences

Thematic area

Biomedical sciences

Status of the project

New

Research proposal

Various immunotherapeutic approaches have been used for the treatment of cancer. Today the most common therapies for malignant diseases are radiotherapy and hemotherapy, although they have severe side effects, due mostly to their non specific action. Different types of cancer need the development of new specific anti-cancer agents targeting cancer cells only without side effects.
The function of tumour-associated macrophages (TAMs) in different types of cancer patients is multifaceted and the literature shows conflicting roles. Many groups tried to characterize the M1 and M2 macrophage populations within TAMs in different subtypes of tumors compared to non-tumour tissue to ascertain their possible inflammatory or immunoregulatory role in the tumor microenvironment.
The differentiation of monocyte-derived-macrophages (MDMs) from the whole blood of healthy donors allows to test the immunoregulatory properties of a selected group of Mediterranean Plant Extracts on MDMs, both alone or in co-culture with tumor cells. The antioxidant, antimicrobial and antiproliferative activities of many Mediterranean plants have been shown in different studies, and we want to extend this analysis to melanoma and glioblastoma human tumors.
Experimental therapy with newly developed drugs in humans is limited by technical and ethical restrictions. Understanding the functional characteristics in human disease where animal models fail to adequately replicate the human condition is a challenging task. Investigating the clinical importance of pathological hallmarks of human melanoma and glioma as tumor invasion in surgical biopsies has been limited to descriptive studies. Studies in humanized mouse models can circumvent some of these limitations.
Severe combined immunodeficient (SCID) mice with T and B immunodeficiency syndrome are a model that is suitable for transferring of human cells without rejection and are unable to mount an adaptive immune response. SCID mice are perfect recipients and reconstituted with human tumor cells develop a symptoms specific for human diseases.

In this collaborative project, we like to establish the animal models of human melanoma and glioblastoma in humanized SCID mice to study the pathogenesis of the diseases. Transferred SCID mice are unique models for immunological investigations. Groups of melanoma or glioblastoma reconstituted SCID mice will be transferred using monocyte-derived-macrophages treated or not with mediterranean plant extracts. The effect of the therapy will be evaluated at the following parameters: tumor growth, survival of the animals and generation of specific cytotoxicity for the melanoma or glioblastoma cells. We expect that after treatment of the melanoma or glioblastoma-bearing experimental animals with treated-MDM the survival of the mice and many of the observed parameters will be improved.
The project team has a strong background in original approaches for specific therapy of murine and human prototype diseases. We are well experienced in clinical research and in establishing precise animal models of murine and human diseases. These skills and knowledge will be applied in the current proposal using various classical and newly developed molecular biology methods and protocols.

The group of Associate Professor Andrey Tchorbanov from the Department of Immunology, The Stefan Angelov Institute of Microbiology of the Bulgarian Academy of Sciences has experience in constructing hybrid antibody molecules by genetic and by protein engineering and humanized SCID - SLE, Allergy, cancer and other murine models and in vitro immunological studies.

The group of Dr. Francesca Mariani, from National Research Council, Institute of Cell Biology and Neurobiology has experience in studying the MDMs innate immune response to infections, with particular respect with Mycobacterium tuberculosis, and had previous collaborations with a Bulgarian group of microbiologists (NCIPD).
This group has recently started to analyse the immunomodulatory properties of Mediterranean Plant extracts (as Lavandula angustifolia, Cinnamon zeylanicum, Crocus sativus, etc.)

The expertise of both laboratories is complementary. We believe this collaborative study could bring productive results in the future.

Research goals

1. Establishment of animal models of tumor diseases in humanized SCID mice.
Humanized melanoma or glioblastoma model development. Immunodeficient SCID mice will be used for this set of experiments for reconstitution of melanoma or glioblastoma cancer cell lines.
2. MDMs differentiation from a first group of five healthy donors whole blood and plants extract treatment of MDMs.
Five healthy donors whole blood buffy coats will be obtained by the University Policlinic Umberto I Transfusion Centre, and subjected to Ficoll gradient to separate Peripheral Blood mononuclear cells (PBMCs). Each of the five MDMs culture will be treated with Lavandula angustifolia, Cinnamomun zeylanicum, and Crocus sativus hydroalcoholic extracts and their M1-M2 phenotype will be analysed.
3.Application of monocyte-derived-macrophages for the pathological response in humanized SCID models of cancer diseases.
a. SCID mice reconstitution with glioblastoma or melanoma cells.
The immunomodulatory activity of MDM will be tested in an in vivo experimental model of human cancer diseases.
b. The effect of the therapy will be evaluated at the following parameters: tumor growth, the effect on the morphological characteristics (size and morphology of the solid tumors), survival of the animals and generation of specific for the melanoma or glioblastoma cells cytotoxicity.
The animal experiments from the second year to be continued for final evaluation of the treatment effect of humanized SCID mice with MDM.

Last update: 06/06/2025