Master thesis project: Validation of miRNA EV-cargo and their impact on AML cells

01.07.2024, Abschlussarbeiten, Bachelor- und Masterarbeiten

Background and Aim: Exosomes are a subtype of extracellular vesicles (EVs) that secreted from cells into the extracellular environment and are involved cell-cell communication, immune responses and tumor progression. The Hecker lab has established a protocol to isolate and characterize extracellular vesicles from hematopoietic stem and progenitor cells (HSPC), from patients with acute myeloid leukemia (AML) and from AML cell lines. Analysis of the cargo of these EVs with RNA-Seq showed a drastic difference in the miRNA content, showing the presence of miRNAs involved in regulation of apoptosis in EVs from AML patients compared to vesicles derived from healthy aged matched individuals. Moreover, treatment of AML cell lines with EV release inhibitors (e.g. FTY720) showed an increased in apoptotic events, potentiated when combined with AML standard of care drugs, such as Venetoclax. Therefore, Hecker lab aim to understand the molecular mechanism leading to the increase in apoptotic event after EV release inhibitor treatment, investigating the role of the miRNA cargo of the EVs.

Methodology: The master student will validate the miRNA cargo data from RNA-Seq via qPCR. Moreover he/she will conduct bioinformatics analysis to find putative targets for the different miRNAs. The putative miRNA targets will be validate in vitro via luciferase assay, upon cloning of the putative gene seed region in the appropriate luciferase plasmid. Moreover, the differential expression of the different miRNAs will be evaluated before and after the treatment with EV inhibitors. The student will be trained in cell culture techniques, exosome isolation and characterization, cell transfection, flow cytometry, RT qPCR and Western Blot. Additionally the student will get insight in cloning strategy and plasmid isolation.

For more information or to apply, please contact the lead of the project, Dr Med Judith Hecker (judith.hecker@mri.tum.de) or Dr Mariagiovanna Barresi (mg.barresi@tum.de).

Kontakt: mg.barresi@tum.de

Mehr Information

https://med3.mri.tum.de/de/ag-dr-med-judith-s-hecker-%E2%80%93-evolution-blood-cancer-0