Project description:Mice with colonic tumors (chemically induced by AOM/DSS) were treated with Nutlin or control vehicle solution to analyze p53 target gene expression in vivo.
Project description:The heat-shock protein 90 (HSP90) is a promising target in cancer therapy, but its inhibitors' clinical trial failures are partly due to a compensatory heat-shock response (HSR) mediated by heat-shock factor 1 (HSF1). We previously showed that wildtype p53 reduces HSR by repressing HSF1 via a p21-CDK4/6-MAPK-HSF1 axis. Here, we explore if simultaneous p53 activation or cell cycle inhibition can disrupt the HSF1-HSR axis and enhance HSP90 inhibitors' efficiency. mRNA sequencing was performed on HCT116 cells treated for 24 hours with DMSO, 50 nM Ganetespib, 1 µM RG-7388, or their combination. We found that the p53 activator Idasanutlin suppresses HSF1-HSR activity in HSP90 inhibitor-based therapies, synergistically reducing cell viability and accelerating cell death in p53-proficient colorectal cancer (CRC) cells and organoids. Combination therapy upregulates p53 pathways, apoptosis, and inflammation. In a CRC mouse model, dual HSF1-HSP90 inhibition represses tumor growth and alters immune cell composition. CDK4/6 inhibition under HSP90 inhibition mimics HSR repression in p53-proficient CRC cells. In p53-deficient CRC cells and p53-mutated organoids, combined HSP90 and CDK4/6 inhibition suppresses HSF1-HSR and reduces cancer growth, offering a p53-independent strategy for CRC treatment. In conclusion, we present new options to improve HSP90-based therapies for enhanced CRC treatment.
Project description:MOLM-13 acute myeloid leukemia cells were treated with 7 µM 5-Azacytidine (Cayman Chemical 11164), or 450 nM CB-5083 (Cayman Chemical 19311), or in combination, or 0.1% DMSO as control. Treatments were conducted for 48 hours.
Project description:Prenatal irradiation can cause neurodevelopmental defects which are characterized by a reduction in brain size (microcephaly). The underlying molecular mechanisms in humans have so far not been studied. Here, we leveraged human forebrain organoids as a model for human embryonic brain development to investigate time- and dose-dependent effects of radiation on organoid growth. For this, organoids of 14 days and 56 days old were irradiated with acute X-ray doses of 0.5 Gy or 2 Gy and compared to controls. Using bulk RNA-seq at different early (6 h and 24 h) and late (14 days) time points after irradiation, we investigated mechanisms of radiation-induced growth defects which resulted from activation of the DNA damage response (cell cycle arrest, DNA repair, apoptosis), premature differentiation and the coordinated repression of primary microcephaly genes.
Project description:K562 cells with a CRISPR-engineered t(7;12)(q36;p13) translocation (K562-t(7;12)) and control line transiently transfected with Cas9 only (K562-ctrl) were FACS sorted on positivity to surface markers Kit and CD24.
Project description:Mice of the genetic background VBPN, VBPNA, VBPNC were injected with tamoxifen systemically (VBPN) or with 4-OHT intra-colonically (VBPNA, VBPNC). After tumor development, the material was processed and RNA-sequencing was conducted.
Project description:In the present study, Marchantia polymorpha Mppcs loss of function mutants were generated through CRISPR/cas9 mediated genome-editing. To assess whether the knockout of MpPCS gene affects the transcription of M. polymorpha nuclear genes in unstressed condition, the Mppcs-2 knockout mutant and Cam2 wild-type transcriptomes were compared by RNA-Seq.
Project description:This dataset identifies genes expressed in florally induced meristems of Col-0 and skrt-2 plants, in control conditions and during salt stress. It allows us to identify how skrt-2 and salt affect the timing of the floral transition, and how the floral transition itself is regulated.