Projects / Programmes
Patient-derived tumour modelling in vitro: induced pluripotent stem cells and 3D organotypic model of disease
| Code |
Science |
Field |
Subfield |
| 4.06.00 |
Biotechnical sciences |
Biotechnology |
|
| Code |
Science |
Field |
| 3.04 |
Medical and Health Sciences |
Medical biotechnology |
patient-derived cancer model, in vitro cancer model, primary cell culture, induced pluripotent stem cells, iPSC, technology, dedifferentiation, 3D organotypic disease model, spheroid, bioengineering
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
35360 |
PhD Sandra Ropret |
Biochemistry and molecular biology |
Head |
2020 - 2022 |
15 |
Organisations (1)
Abstract
ABSTRACT Worldwide, cancer is the second leading cause of mortality and, according to the latest available data, around 9.6 million people have died from cancer in 2018. Similarly to global situation, cancer is the second leading cause of mortality in Slovenia. Cancer represents a group of heterogeneous and complex diseases characterized by the uncontrolled proliferation of transformed cells. Risk factors for cancer development include environment, lifestyle, hereditary predisposition, and also coincidence. Among cancers only 20 percent are hereditary, while 80 percent of cancers occur randomly, meaning that clear causes for the disease development are unknown The most commonly used model systems in basic cancer research are immortalized cell lines and animal models. Immortalized cell lines are inexpensive and simple two-dimensional (2D) model. However, a growing body of experimental evidence shows that the relevant features of the cancer progression, and the treatment effectiveness of anticancer compounds, cannot be properly recapitulated in such model. On the contrary, animal models satisfy the tri-dimensionality (3D) requirement, and enable in vivo exploration of tumour growth, behaviour, and treatment effectiveness. However, animal models are time-, resource-, and expertise-intensive, and they fail to provide a precise recapitulation of human physiology. Moreover, the animal use is increasingly being questioned by different public entities for ethical concerns. Shortcomings of models often result in a low translation rate of the findings of basic research into the clinical environment. Nowadays, patient-derived models (e.g. 3D cell culturing, patient-derived xenografts, organs-on-chips) are gaining in popularity. However, the quantity of biological samples from patients is limited. The induced pluripotent stem (iPS) cell technology, pioneered in 2006 by Shinya Yamanaka and Kazutoshi Takahashi, may circumvent this quantity limitation. The technology enables reprogramming of somatic cells to pluripotency by transient exogenous expression of a defined set of transcription factors. The obtained iPS cells have the capacity of unlimited proliferation and self-renewal, and potential to differentiate into all three germ layer cells. Thus, the technology may provide renewable sources of biologically relevant cells. In conjunction with robust differentiation protocols, iPS cells generated from human primary malignant cells provide opportunity for in-depth exploration of pathophysiology and treatment of human cancer. Moreover, when combined with 3D cell culturing approaches, the iPS cells offer an opportunity for establishment of in vitro cancer models that closely resemble human tumour in vivo situation. First objective of the proposed research is derivation of fully reprogrammed iPS cells from primary breast cancer cells. The reprogramming will be done by non-integrating approach, and the reprogramming protocol will be tailored to the cancer type. The iPS cells will be initially generated from commercially obtained primary tumour cells. Next, the iPS cells will be generated from primary tumour cells of Slovenian breast cancer patient. Since the chosen malignancy is nonhematologic, the normal iPS cells with the same genetic background (i.e. isogenic iPS cells) will be generated from preipheral blood cells of the same Slovenian breast cancer patient. The obtained iPS cells will be quality-controlled for the absence of transgenes (qPCR), and presence of pluripotency and trilineage differentiation markers on mRNA and protein level (qPCR, immunocitchemistry). Second objective is generation of a 3D breast cancer model (spheroids), from the patient-derived iPS cells, using scaffold-free hanging drop technique. The generated iPS cells and spheroids can further serve as an expandable tool for in vitro exploration of e.g. cancer evolution, cancer cell plasticity, development/testing of cancer type specific drugs.
