TY - THES A1 - Koike, Akito T1 - Molekular und zellbiologischer Ansatz hin zu neuartigen Medikamenten gegen \(Echinococcus\) \(multilocularis\) T1 - Molecular and cell biological approach towards novel drugs against \(Echinococcus\) \(multilocularis\) N2 - Echinococcosis is an important zoonosis. The causative agent of Alveolar Echinococcosis (AE) is Echinococcus multilocularis. The treatment of human AE is limited to surgery and chemotherapy with albendazole (ABZ). However, ABZ works only parasitostatically and it needs to be taken for long periods, although it causes adverse side effects. Thus, development of new, parasiticidal drug with selective toxicity is required. Because undifferentiated stem cells of E. multilocularis play key role in its longevity and regenerative capacity, targeting stem cells is especially important. In vitro screening of protein kinases inhibitors demonstrated that human PIM kinases inhibitors have detrimental effects on E. multilocularis. Through yeast two hybrid assay, the interaction of parasite PIM kinase (EmPIM) and its CDC25 (EmCDC25) was indicated. Through in situ hybridization, expression of EmPIM in the stem cells was observed. Therefore, EmPim is likely to be a positive regulator of cell cycle progression, the same as human Pim1. In addition, 20 compounds against EmPIM were selected through in silico screening and synthesized. One of them has a detrimental effect on E.multilocularis comparable to human pan-PIM inhibitors, but has much weaker toxicity on human cell lines. Furthermore, triclabendazole (TCBZ) and its metabolite TCBZSX, which are approved for another flatworm disease, Fascioliasis were tried on E. multilocularis. With two stem cell markers, damage to stem cells by TCBZSX was shown. In addition, primary cells from treated vesicles never regenerated and the damage to stem cells proved to be irreversible. Our in silico screening method used in EmPIM research has potential to identify compounds which overcome the side effect problem in ABZ-based chemotherapy. On the other hand, it is expected that my research of TCBZ can lead to development of a practical parasiticidal chemotherapy by combining TCBZ, which damages stem cells, and ABZ, which damages differentiated cells. N2 - Die Echinokokkose ist eine der wichtigsten Zoonosen sowohl für die Human- als auch für die Veterinärmedizin. Der Erreger der alveolären Echinokokkose (AE) ist Echinococcus multilocularis. Metazestode Bläschen, das Larvenstadium dieses parasitären Helminthen, können in die Leber eindringen und ungeschlechtlich wie bösartige Tumore wachsen. Dies kann ohne geeignete Behandlung tödlich sein. Die Behandlung von AE beim Menschen beschränkt sich auf Chirurgie und Chemotherapie, aber die Chirurgie ist nur bei einem kleinen Prozentsatz der Patienten anwendbar, die im Frühstadium diagnostiziert werden. Die meisten Patienten können sich nur auf eine Chemotherapie mit Albendazol (ABZ) verlassen. ABZ wirkt jedoch nur parasitostatisch und kann die Krankheit nicht heilen. Daher muss ABZ über einen längeren Zeitraum eingenommen werden, obwohl es mit Nebenwirkungen einhergeht. Daher ist die Entwicklung eines neuen, parasitentötenden und selektiven Medikaments gegen AE erforderlich. Da die undifferenzierte Stammzellpopulation von E. multilocularis eine Schlüsselrolle für seine Langlebigkeit und Regenerationsfähigkeit spielt, ist eine auf Stammzellen abzielende Chemotherapie wichtig. In dieser Arbeit wurde ein In-vitro-Screening verschiedener Hemmstoffe gegen Kinasen und Tubuline durchgeführt. Das Ergebnis des Screenings zeigte, dass Inhibitoren gegen humane pim-Kinasen starke schädliche Auswirkungen auf E. multilocularis haben. Durch ein Hefe-Zwei- Hybrid-System wurde die Interaktion der Parasiten-Pim-Kinase (EmPIM) mit der Zellteilungszyklus 25 (EmCDC25) nachgewiesen, und durch In-situ-Hybridisierung wurde die teilweise Lokalisierung von EmPIM in den Stammzellen beobachtet. Daher ist es wahrscheinlich, dass EmPim ein positiver Regulator der Zellzyklusprogression ist, genau wie menschliches Pim1. ... KW - Bandwürmer KW - Zellzyklus KW - Benzimidazolderivate KW - tapeworm KW - kinase KW - benzimidazole Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-288649 ER - TY - JOUR A1 - Cucher, Marcela A. A1 - Mariconti, Mara A1 - Manciulli, Tommaso A1 - Vola, Ambra A1 - Rosenzvit, Mara C. A1 - Brehm, Klaus A1 - Kamenetzky, Laura A1 - Brunetti, Enrico T1 - Circulating small RNA profiling of patients with alveolar and cystic echinococcosis JF - Biology N2 - Alveolar (AE) and cystic (CE) echinococcosis are two parasitic diseases caused by the tapeworms Echinococcus multilocularis and E. granulosus sensu lato (s. l.), respectively. Currently, AE and CE are mainly diagnosed by means of imaging techniques, serology, and clinical and epidemiological data. However, no viability markers that indicate parasite state during infection are available. Extracellular small RNAs (sRNAs) are short non-coding RNAs that can be secreted by cells through association with extracellular vesicles, proteins, or lipoproteins. Circulating sRNAs can show altered expression in pathological states; hence, they are intensively studied as biomarkers for several diseases. Here, we profiled the sRNA transcriptomes of AE and CE patients to identify novel biomarkers to aid in medical decisions when current diagnostic procedures are inconclusive. For this, endogenous and parasitic sRNAs were analyzed by sRNA sequencing in serum from disease negative, positive, and treated patients and patients harboring a non-parasitic lesion. Consequently, 20 differentially expressed sRNAs associated with AE, CE, and/or non-parasitic lesion were identified. Our results represent an in-depth characterization of the effect E. multilocularis and E. granulosus s. l. exert on the extracellular sRNA landscape in human infections and provide a set of novel candidate biomarkers for both AE and CE detection. KW - echinococcosis KW - small RNA KW - extracellular KW - circulating KW - microRNA KW - serum KW - tapeworm KW - diagnosis KW - marker KW - Echinococcus Y1 - 2023 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-319270 SN - 2079-7737 VL - 12 IS - 5 ER -