Project description:Primary objectives: The primary objective is to investigate circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Primary endpoints: circulating tumor DNA (ctDNA) via deep sequencing for mutation detection and by whole genome sequencing for copy number analyses before start (baseline) with regorafenib and at defined time points during administration of regorafenib for treatment efficacy in colorectal cancer patients in terms of overall survival (OS).
Project description:We used Candida albicans lab strain SC5314 to obtain tunicamycin adaptors. We did whole genome sequencing of the adaptors and the parent as well.
Project description:Candida lusitaniae is an emerging human opportunistic yeast, which can switch from yeast to pseudohyphae, and one of the rare Candida species capable of sexual reproduction. Its haploid genome and the genetic tools available make it a model of interest to study gene function. This study describes the consequences of DPP3 inactivation on cell morphology and mating, both altered in the dpp3Δ knock-out. Interestingly, reintroducing a wild-type copy of the DPP3 gene in the dpp3Δ mutant failed to restore the wild-type phenotypes. Proteomic analyses showed that about 150 proteins were statistically deregulated in the dpp3Δ mutant, and that most of them did not return to their wild-type level in the reconstituted DPP3 strain. The analysis of the segregation of the dpp3Δ mutation and the phenotypes in the progeny of a cross (between the dpp3Δ knock-out and a wild-type strain) showed that the phenotypes are not linked to dpp3Δ, but to a secondary mutation. Genome sequencing of the dpp3Δ mutant allowed us to identify this secondary mutation.
Project description:Miltefosine has been identified for its antifungal properties, yet the mechanisms underlying its action and the development of resistance remain unclear. In this study, we induced drug resistance in various Candida species through concentration gradient exposure, successfully obtaining a miltefosine-resistant strain of Candida glabrata. Whole-genome sequencing revealed a premature stop codon mutation in the OSH2 gene within the resistant strain. To further investigate the function of the Osh2, we deleted it in Candida albicans, observing a significant increase in miltefosine resistance in the knockout strain. RNA-seq and lipidomics analyses indicated that Osh2 influences the ergosterol biosynthetic pathway by regulating zymosterol transport, thereby mediating miltefosine resistance. Additionally, exogenous supplementation of zymosterol significantly enhanced the resistance of Candida to miltefosine. Further studies revealed that deletion of the ERG11 gene, a rate-limiting enzyme in the zymosterol synthesis pathway, rendered the strain highly sensitive to miltefosine, while deletion of the ERG6 gene, which catalyzes the conversion of zymosterol to fecosterol, led to miltefosine resistance. This study is the first to elucidate that the Osh2 gene mediates miltefosine resistance in Candida by regulating zymosterol transport and affecting the ergosterol biosynthetic pathway, providing new theoretical insights into the mechanism of action of miltefosine and strategies to overcome fungal resistance.
Project description:We treated Candida albicans lab strain SC5314 with fluorouracil and we did whole genome sequencing of 24 adaptors as well as the parent
Project description:We exposed Candida parapsilosis clinical isolate #12108 to YPD plate supplemented with 8µg/ml of tunicamycin. We randomly selected 18 adaptors. We did sequencing of these adaptors.