1.1.1.1/18/A/089
„RNA molecular determinants in development of pituitary adenoma”
Funding: European Regional Development Fund (ERDF), Measure 1.1.1.1 “Support for applied research”
Project No.: 1.1.1.1/18/A/089
Period: 1 April 2019 – 31 March 2022
Project costs: 648 648 EUR
Principle Investigator: Dr. biol. Vita Rovīte
Cooperation partner: SIA “GenEra”
Project summary:
The scientific objective of the project is to study the spectrum of various ribonucleic acid (RNA) markers (mRNA, non-coding) in development of pituitary adenoma (PA) to discover determinants that influence therapy response and disease outcome.
The main activities to achieve the goal of the project: 1) identify and analyse molecular RNA factors in primary PA tissue by massive parallel sequencing (MPS), 2) study RNA types in primary culture of free-floating spheres and mesenchymal cells from PA tissue to describe tumour-borne RNA patterns, 3) investigate alterations in coding and non-coding RNA in free-floating sphere model upon addition of PA treatment medication, 4) validate discovered markers in PA patients’ plasma and independent cohort, 5) functionally characterize discovered RNA candidates in GH3 cells.
The results of the project will evaluate molecular RNA factors on several functional levels of PA derived biological materials and models: (1) potential makers discovered in tissues and validated in plasma and/or tissues could serve for prediction of disease outcome, characterization of aggressiveness and non-invasive diagnostics, (2) study of free-floating spheres and mesenchymal cells will help to estimate spheres as isolated model for PA tumour-borne cells, define their RNA profile and describe PA therapy response in this system, (3) research of influence of discovered RNA candidates in GH3 cells will functionally characterize the precise cellular mechanisms of these factors in PA development.
Information published 01.04.2019.
Progress of the project:
1 April 2019 – 30 June 2019
The project specific sample database has been created, data was gathered about tissue from surgery material, phenotypic information of the patients has been gathered, sorted and quality controlled. DNA and RNA from newly acquired tissue samples have been carried out, and list of samples for further RNA sequencing analysis was created. The cell lines of mesenchymal cells and spheres from newly acquired primary adenoma tissue have been established and cell-derived RNA materials have been obtained for further project implementation tasks.
Information published 28.06.2019.
Progress of the project:
1 July 2019 – 30 September 2019
The creation of project specific sample database has been continued, data was gathered about tissue from surgery material, phenotypic information of the patients has been gathered, sorted and quality controlled. DNA and RNA from newly acquired tissue samples have been carried out, and first RNA sequencing of PA tissue have been performed. The cell lines of mesenchymal cells and spheres from newly acquired primary adenoma tissue have been established and incubated with octreotide and cabergoline, cell-derived RNA materials have been obtained for further project implementation tasks.
Information published 30.09.2019.
Progress of the project:
1 October 2019 – 31 December 2019
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional five pituitary adenoma tissue samples have been obtained, one part of the tissue was used for DNA and RNA isolation and other part have been used for the development of mesenchymal cells and sphere cultures, that were incubated in octreotide and cabergoline for further project implementation tasks. The RNA next generation sequencing library preparation, quality control and sequencing have been continued.
Information published 30.12.2019.
Progress of the project:
1 January 2020 – 31 March 2020
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional two pituitary adenoma tissue samples have been obtained, one part of the tissue was used for DNA and RNA isolation and other part have been used for the development of mesenchymal cells and sphere cultures, that were incubated in octreotide and cabergoline for further project implementation tasks. The RNA next generation sequencing library preparation, quality control and sequencing have been continued. Data analysis protocol for small non-coding RNA has been optimized.
Information published 31.03.2020.
Progress of the project:
1 April 2020 – 30 June 2020
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional three pituitary adenoma tissue samples have been obtained, one part of the tissue was used for DNA and RNA isolation and other part have been used for the development of mesenchymal cells and sphere cultures, that were incubated in octreotide and cabergoline for further project implementation tasks. The transcriptome and small non-coding RNA next generation sequencing library preparation, quality control and sequencing have been continued. The primary data bioinformatics analysis have been started.
Information published 30.06.2020.
Progress of the project:
1 July 2020 – 30 September 2020
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional two pituitary adenoma tissue samples have been obtained, one part of the tissue was used for DNA and RNA isolation and other part have been used for the development of mesenchymal cells and sphere cultures, that were incubated in octreotide and cabergoline for further project implementation tasks. The transcriptome and small non-coding RNA next generation sequencing library preparation, quality control and sequencing have been continued. The data bioinformatics analysis has been continued for transcriptome and small non coding RNA data.
Information published 30.09.2020.
Progress of the project:
1 October 2020 – 31 December 2020
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional three pituitary adenoma tissue samples have been obtained, one part of the tissue was used for DNA and RNA isolation and other part have been used for the development of mesenchymal cells and sphere cultures, that were incubated in octreotide and cabergoline for further project implementation tasks. The small non-coding RNA next generation sequencing library preparation, quality control and sequencing have been continued. The optimization of transcriptome sequencing of spheroids has been carried out, transcriptome sequencing of primary tumour tissues has been finished and bioinformatic analysis is ongoing. The functional experiments have been started in cell lines.
Information published 30.12.2020.
Progress of the project:
1 January 2021 – 31 March 2021
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional one pituitary adenoma tissue sample has been obtained and processed for project implementation. The detailed bioinformatics analysis of sequencing data is ongoing in several research segments (tumour and cell line transcriptome, small noncoding miRNA). The miRNA marker validation in independent sample group is ongoing as well as functional experiments for detailed tumour development cellular mechanism characterization in cell lines.
Information published 31.03.2021.
Progress of the project:
1 April 2021 – 30 June 2021.
The additional data entrance in the project specific sample and clinical information database has been continued. In this implementation period additional two pituitary adenoma tissue samples have been obtained, nucleic acids have been isolated and the rest of the tissue used for cell line obtainment. The detailed bioinformatics analysis of sequencing data is ongoing for tumour and cell line transcriptome and small noncoding miRNA. The miRNA marker validation in independent sample group is ongoing as well as functional experiments for detailed tumour development cellular mechanism characterization in cell lines.
Information published 30.06.2021.
Progress of the project:
1 July 2021 – 30 September 2021
The additional data entrance in the project specific sample and clinical information database has been continued. The detailed bioinformatics analysis of sequencing data has been completed for tumour and cell line transcriptome and small noncoding miRNA. Currently, thorough interpretation of obtained results is ongoing and the data are prepared for manuscript preparation. The miRNA marker validation in independent sample group is ongoing as well as functional experiments for detailed tumour development cellular mechanism characterization in cell lines.
Information published 30.09.2021.
Progress of the project:
1 October 2021 – 31 December 2021
The additional data entrance in the project specific sample and clinical information database has been finished. The thorough interpretation of obtained results from tumour, tumour cell lines and small non-coding RNA sequencing are ongoing and prepared for manuscript preparation. The miRNA marker validation in independent sample group is ongoing as well as functional experiments for detailed tumour development cellular mechanism characterization in cell lines.
Information published 30.12.2021.