Prolonged hypoxia induced differential protein signatures in BRCA1 +/- triple negative breast cancer cell lines

Breast cancer is the most common cancer in Europe (age standardized incidence 109 cases per 100 000 population, accordingly, in 2012), and results in substantial socioeconomic burden on the society. Triple-negative breast cancer (TNBC) is associated with a poor prognosis and defines a subgroup of patients who do not benefit from endocrine or anti-HER2 therapy. Although TNBC has relative chemo-sensitivity it is still characterized by aggressive clinical behaviour. In our previous studies we have proved that relapse rates and breast cancer-specific survival (BCS) outcomes for BRCA1 positive (+) TNBC is significantly better compared to BRCA1 negative (-) TNBC. This finding is in the line with proposed involvement of altered DNA double strand break (DSB) repair system in therapy response.

Proposed project aims to analyse differential protein biosynthesis in BRCA1 +/- TNBC cell lines, cultured under prolonged hypoxia (to mimic in vivo microenvironment of breast cancer). Identified differentially expressed proteins will be analysed in respect to DSB repair signalling pathways to identify potential therapy targets for this aggressive type of breast cancer.

1. Protein isolation from cultured cell lines – RSU Institute of Oncology.
2. Protein concentration determination and quality assessment with 1D gel electrophoresis – RSU Institute of Oncology.
3. Sample preparation for mass-spectrometry – RSU Institute of Oncology.
4. Sample analysis with LC-MSe/HDMSe – IZKF Core Unit Proteomics, WWU Munster.
5. Data analysis with PLGS and bioinformatics – IZKF Core Unit Proteomics, WWU Munster (together with two scientists from Latvia team who will visit Munster).
6. Identified proteins pathway analysis – RSU Institute of Oncology
7. Manuscript / conference thesis preparation for publication of acquired results – RSU Institute of Oncology in cooperation with IZKF Core Unit Proteomics, WWU Munster.

Direct:

The scientific group (3 scientists from RSU IO) involved in the proposed project will gain experience in proteomics and data analysis. The acquired knowledge in proteomics will open up the possibility of expanding the field of research, strengthen cooperation established during this project and find new collaboration partners for whole scientific personnel of Institute of Oncology (20 scientists).

Indirect:

The gained experience will contribute to cooperation between departments within Riga Stradiņš University (RSU), such as Department of Biology and Microbiology, and Department of Surgery etc. The successful implementation of this project will benefit to the improvement of the innovation potential and scientific competitiveness of RSU. The new knowledge base created within the scopes of this project will be transferred to fulfil requirements for up-to date education for medical students in pre-clinical courses (such as Molecular biology, 1^st year students, ~ 500) and clinical courses (conducted by Department of Surgery, 4^th and 5^th year students).

Potential users of project results will be health care system and medical professionals – surgeons, oncologist-chemotherapists, who are involved in the care of breast cancer patients.

The scientific and collaborative experience will be shared within Institute of Oncology.

The project results among medical professionals will be disseminated through professional societies. The results of project will be discussed among geneticists and molecular biologists during the meeting of Latvian Association of Human Genetics (planed in September, 2019), presenting author Zanda Daneberga.