Dolzany de Oliveira, Thaís ORCID: 0000-0002-5503-4051 (2023). Unraveling the Role of Lyn Kinase in the Extracellular Vesicle-Based Crosstalk Between Primary Chronic Lymphocytic Leukemia Cells and Stromal Cells. PhD thesis, Universität zu Köln.
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Abstract
In Chronic Lymphocytic Leukemia (CLL), the tumor cells receive decisive survival support from non-malignant cells within the bone marrow. This support is stimulating tumor growth, progression and helps leukemic cells to evade chemotherapy. Within this microenvironment, the crosstalk between stromal and tumor cells plays an important role. The cell types interact directly and through the paracrine release of intercellular messengers, such as growth factors, and released plasma membrane-enclosed extracellular vesicles (EVs). In particular, the communication by EVs is not fully understood. It has already been shown that members of the Src family of kinases (SFKs) can influence the EV secretion and the SFK member Lyn is aberrantly expressed and highly active CLL-affected tissue, not only on malignant CLL cells but also in macrophages and stromal cells. This thesis aimed to study the mechanism of the Src kinase Lyn in the EV-based communication between the malignant B-CLL and stromal cells. Comparing Lyn proficient and deficient stromal cell lines revealed that the lack of Lyn raised the cell adhesion and podosome formation but decreased the number of filopodia per cell. Moreover, Imaging Flow cytometry and nanoparticle tracking analysis revealed that the EV release and EV uptake are significantly reduced in Lyn deficient stromal cells as compared to wild-type (WT) counterparts, resulting in a 36% reduction of the EV release and a 16% reduction in EV uptake, indicating that Lyn influences both, the cellular release and uptake of EVs. In addition, the same amount (4 μg/ml) of EVs from Lyn proficient stromal cells induced significantly higher support on primary CLL cells as compared to EVs from Lyn deficient counterparts. These data suggest that Lyn not only influences the EV release but also the molecular composition of the EVs. Proteomic comparison of the Lyn proficient and the deficient stromal cell line HS5 highlighted 72 significantly differentially expressed proteins. Among them, CD248 was prominently decreased in Lyn-deficient HS5 cells and a knockdown of CD248 in HS5 cells resulted in a diminished B-CLL cells survival feeding capacity compared to WT cells. In conclusion, the presented data provide initial preclinical evidence, that the tyrosine kinase Lyn crucially influences the EV-based communication between primary B-CLL and supporting bystander cells by raising the EV release and their concentration of functional molecules, such as CD248.
Item Type: | Thesis (PhD thesis) | ||||||||||||||
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URN: | urn:nbn:de:hbz:38-734932 | ||||||||||||||
DOI: | 10.3389/fmed.2022.1059028 | ||||||||||||||
Date: | 2023 | ||||||||||||||
Publisher: | Frontiers in Medicine | ||||||||||||||
Language: | English | ||||||||||||||
Faculty: | Faculty of Mathematics and Natural Sciences | ||||||||||||||
Divisions: | CECAD - Cluster of Excellence Cellular Stress Responses in Aging-Associated Diseases | ||||||||||||||
Subjects: | Natural sciences and mathematics Medical sciences Medicine |
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Date of oral exam: | 23 June 2022 | ||||||||||||||
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Funders: | DFG | ||||||||||||||
Refereed: | Yes | ||||||||||||||
URI: | http://kups.ub.uni-koeln.de/id/eprint/73493 |
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