OCIC · Ongoing chromosomal instability in cancer: real-time imaging and single cell genetics of missegregating chromosomes.
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2021-01-01 → 2022-12-31
- EU contribution
- €187,572
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Ongoing chromosomal instability in cancer: real-time imaging and single cell genetics of missegregating chromosomes.
While many cancer treatments are improving, cancer remains a major cause of death and suffering, affecting both patients and their families. Increased life expectancy has further increased cancer incidence, putting a heavy burden on society,increasing pressure and healthcare and costs. The objectives of this project are to add fundamental knowledge on how genetic instability is related to ongoing tumor development. DNA analysis of patient tumors show many different genomic alterations, ranging from point mutations, gene amplifications/losses, to chromosomal rearrangements such as structural variations. Especially gross chromosomal aberrations involving whole chromosomes or are likely to occur during mitosis, a crucial phase in the cell cycle at which the DNA needs to be correctly distributed over both daughter cells. The fundamental principles of cell division have been studied in different model systems, invaluable in untangling the genetic components, signaling pathways and timing of the components that orchestrate correct cell division. However, to study cell division in a model system that is most representative for human tumors, including 3 dimensional growth and heterogeneity, compatible with high resolution live-cell imaging, we studied patient derived tumor organoids (PDOs). This model allows to study chromosomal instability (CIN), focusing on chromosomal missegregations and mitotic errors during tumor cell proliferation. Tumors of esophageal origin, such as esophageal adenoma carcinoma’s (EAC) are of specific interest to study the impact of CIN, since EACs seem not driven by sequential acquisition of a specific driver mutations. Rather, EACs progress through multiple combinations of chromosomal alterations, including copy-number changes and large structural variants.
Data: CORDIS, © European Union
Project objective
Genomes from tumor cells are often grossly derailed in DNA content, with chromosomes gained, lost, fused or otherwise rearranged. Presumably, many of these aberrations stem from mistakes during cell division, during which the DNA is normally divided equally to both daughter cells. Cell division is often studied microscopically in 2D cell culture, while DNA sequences from cancer genomes are often re-constructed from tumor patient material. As a consequence, there is still a poor understanding regarding the direct genetic consequences of mitotic errors and as such, genomic instability. Patient-derived tumor organoids, in combination with cutting-edge technology that combines direct microscopic observation of mitotic events, with single-cell whole genome sequencing (scWGS), provide a unique opportunity to tackle these fundamental questions.To understand the occurrence and maintenance of genomic instability, I will study organoids derived from esophageal adenocarcinoma (EAC) patients. While observed in many cancers, EAC genomes in particular show genomic catastrophes and are thought to drive tumorigenic transformation. Cells will be filmed in real-time to detect mitotic errors. Daughter cells from erroneous divisions will be photoconverted, allowing single cell tracking and isolation for prospective genetic analysis. scWGS will detect imbalances in chromosome number and/or structural rearrangements between the paired daughter cells. Genetic aberrations can be directly linked to the observed mitotic error and cell fate across multiple divisions to understand its role in tumorigenesis. To couple molecular mechanisms to genetic footprints, fluorescent markers will be implemented to interrogate the type of DNA damage missegregated chromosomes receive, how this affects the subsequent cell cycles, and how this missegregated DNA is re-incorporated into the genome. Finally, I will test these findings across a panel of human tumor organoids derived from different tissues.
Original text from CORDIS.
Participants
- UNIVERSITAIR MEDISCH CENTRUM UTRECHT · UtrechtCoordinatorNetherlands
Links
Data: CORDIS, © European Union