Descrizione del progetto
Studio in tempo reale sull’instabilità cromosomica nel cancro
Molte aberrazioni genomiche hanno origine da errori durante la divisione cellulare, in cui il DNA viene di norma suddiviso equamente tra entrambe le cellule figlie. Sono tuttora scarsamente comprese le conseguenze genetiche degli errori mitotici per la tumorigenesi e l’instabilità genomica. Organoidi tumorali derivati dal paziente e tecnologie di ricerca all’avanguardia, che combinano l’osservazione microscopica diretta della mitosi e il sequenziamento a singola cellula dell’intero genoma, forniscono un’opportunità per studiare tali questioni fondamentali. Il progetto OCIC, finanziato dall’UE, studierà l’espressione e il mantenimento dell’instabilità genomica, avvalendosi di organoidi provenienti da pazienti affetti da adenocarcinoma dell’esofago (EAC, Esophageal AdenoCarcinoma) come sistema modello. I genomi dell’EAC, in particolare, mostrano catastrofi genomiche che potrebbero condurre la tumorigenesi. In definitiva, lo studio si propone di applicare la tecnologia a un gruppo di organoidi tumorali umani provenienti da tessuti differenti.
Obiettivo
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.
Campo scientifico
Programma(i)
Argomento(i)
Meccanismo di finanziamento
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinatore
3584 CX Utrecht
Paesi Bassi