Dokument: AI-assisted analysis of flow cytometry data from human hematopoietic cells
| Titel: | AI-assisted analysis of flow cytometry data from human hematopoietic cells | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=67062 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20241021-090343-8 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Nollmann, Cathrin [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof. Dr. Heinzel, Thomas [Gutachter] Prof. Dr. Haas, Rainer [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik » 530 Physik | |||||||
| Beschreibung: | This thesis deals with the investigation of AI-assisted analysis of flow cytometry data from human hematopoietic cells and how this can contribute to clinical diagnostics. In addition, investigations into the establishment of a drug delivery system with carbon nanodots (CNDs) synthesized in the working group were continued. Four manuscripts emerged from this work, which summarize the main findings.
In Paper I, the immunological phenotype of hematopoietic stem and progenitor cells (HSPCs) was analyzed using selected markers in comparison to blast and leukemic stem cells (LSCs) from patients with acute myeloid leukemia (AML) and myelodysplastic neoplasms (MDS) assisted by the t-SNE algorithm. The gates defined on the t-SNE map could be assigned to diagnostically relevant cell populations and revealed a fine substructure within the respective cell types. For the selected markers it has been shown that the differences between HSPCs in comparison to blasts, and LSCs in AML and MDS are likely to be due to the different proportions of certain cell types and different intensities of fluorescent markers within the various cell types, rather than the exclusive presence of certain cell types. In addition, a method was introduced for classifying new samples using t-SNE reference images and a quantitative similarity comparison with the pearson coefficient as a measure. This concept can also be used to monitor the evolution of cell populations of patients in therapy and is not limited to the diagnosis of AML and MDS, but can be applied to multiparameter diagnostic flow cytometry data as well as single-cell data in general. As a further parameter in the AI-supported evaluation of hematopoietic flow cytometry data, the uptake of CNDs was investigated in Paper II in comparison between healthy donors and patients with AML. The differential uptake of CNDs between different cell types as well as between the two comparison groups could be shown concisely using t-SNE. While all cell types took up the CNDs, the CD34+ and CD33+ subsets of the AML samples showed a significantly reduced uptake. Confocal fluorescence microscopy images showed that the CNDs accumulate perinuclearly in the AML cell line HL-60, indicating localization in the lysosomes, in agreement with previous studies. In Paper III, the possibilities of functionalizing the CNDs for selective uptake were investigated. For this purpose, monosaccharides and glycooligomers were covalently coupled to the CNDs and the uptake was investigated in multiple cell lines. No cell type-dependent uptake was observed, but the CNDs conjugated with sugar monomers showed a two- to threefold increase in uptake compared to the pristine CNDs and the CNDs functionalized with glycooligomers. Finally, Paper IV shows the results of the study on the influence of CNDs on the lysosomes and the associated process of autophagy, as well as the targeted delivery of drugs into the lysosomes using CNDs as carriers. Lysosomal processes and autophagy as characterized by cathepsin B and L and the autophagy markers SQSTM1/p62 and LC3 were not significantly altered due to the presence of the CNDs, making them promising inert carriers for lysosomal drug delivery. Branched Polyethylenimine (bPEI) was coupled to the CNDs as an example of a drug. bPEI was successfully delivered into lysosomes by the CNDs, confocal fluorescence microscopy revealing increased accumulation of bPEI-CNDs in lysosomes compared to pristine CNDs. In addition, it was observed that the effects of free bPEI on the cell were attenuated by binding to the CNDs. | |||||||
| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Fachbereich / Einrichtung: | Mathematisch- Naturwissenschaftliche Fakultät » WE Physik » Physik der kondensierten Materie | |||||||
| Dokument erstellt am: | 21.10.2024 | |||||||
| Dateien geändert am: | 21.10.2024 | |||||||
| Promotionsantrag am: | 23.07.2024 | |||||||
| Datum der Promotion: | 26.08.2024 |
