Effective adjunctive therapy by an innate defense regulatory peptide in a preclinical model of severe malaria

Case fatality rates for severe malaria remain high even in the best clinical settings because antimalarial drugs act against the parasite without alleviating life-threatening inflammation. We assessed the potential for host-directed therapy of severe malaria of a new class of anti-inflammatory drugs, the innate defense regulator (IDR) peptides, based on host defense peptides. The Plasmodium berghei ANKA model of experimental cerebral malaria was adapted to use as a preclinical screen by combining late-stage intervention in established infections with advanced bioinformatic analysis of early transcriptional changes in co-regulated gene sets. Coadministration of IDR-1018 with standard first-line antimalarials increased survival of infected mice while down-regulating key inflammatory networks associated with fatality. Thus, IDR peptides provided host-directed adjunctive therapy for severe disease in combination with antimalarial treatment

Red2Flpe-SCON: a versatile, multicolor strategy for generating mosaic conditional knockout mice

Abstract Image-based lineage tracing enables tissue turnover kinetics and lineage potentials of different adult cell populations to be investigated. Previously, we reported a genetic mouse model system, Red2Onco, which ectopically expressed mutated oncogenes together with red fluorescent proteins (RFP). This system enabled the expansion kinetics and neighboring effects of oncogenic clones to be dissected. We now report Red2Flpe-SCON: a mosaic knockout system that uses multicolor reporters to label both mutant and wild-type cells. We develop the Red2Flpe mouse line for red clone-specific Flpe expression, as well as the FRT-based SCON (Short Conditional IntrON) method to facilitate tunable conditional mosaic knockouts in mice. We use the Red2Flpe-SCON method to study Sox2 mutant clonal analysis in the esophageal epithelium of adult mice which reveal that the stem cell gene, Sox2, is less essential for adult stem cell maintenance itself, but rather for stem cell proliferation and differentiation

Red2Flpe-SCON: a versatile, multicolor strategy for generating mosaic conditional knockout mice

Acknowledgements: We thank present and past members of the Koo, Elling and Urban labs at IMBA for valuable discussions and critical comments, Dr. Rike Zietlow and the Life Science Editors for reading and correcting the manuscript, VBC core facilities (especially the Histopathology facility, BioOptics and the animal caretakers). We thank Single Cell Discoveries for helping us with single cell RNA sequencing and offering technical advice on sample handling. This work was supported by core funding from the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences; ERC starting grant, Troy Stem cells, 639050; Interpark Bio-Convergence Center Grant Program; and fellowship to S.W. (DOC Fellowship of the Austrian Academy of Sciences). G.C. was supported by a Lise Meitner Postdoctoral fellowship M2976, FWF, and by the FWF Standalone (P35694) and ERA PerMed (I 5900) grants. B.-K.K. and his team are supported by the Institute for Basic Science.Funder: This work was supported by core funding from the Institute of Molecular Biotechnology (IMBA) of the Austrian Academy of Sciences; ERC starting grant, Troy Stem cells, 639050; Interpark Bio-Convergence Center Grant Program; and fellowship to first author Sam Wu (DOC Fellowship of the Austrian Academy of Sciences)Funder: FWF Lise Meitner Postdoctoral fellowship M2976 FWF Standalone P35694 FWF ERA PerMed I 5900AbstractImage-based lineage tracing enables tissue turnover kinetics and lineage potentials of different adult cell populations to be investigated. Previously, we reported a genetic mouse model system, Red2Onco, which ectopically expressed mutated oncogenes together with red fluorescent proteins (RFP). This system enabled the expansion kinetics and neighboring effects of oncogenic clones to be dissected. We now report Red2Flpe-SCON: a mosaic knockout system that uses multicolor reporters to label both mutant and wild-type cells. We develop the Red2Flpe mouse line for red clone-specific Flpe expression, as well as the FRT-based SCON (Short Conditional IntrON) method to facilitate tunable conditional mosaic knockouts in mice. We use the Red2Flpe-SCON method to study Sox2 mutant clonal analysis in the esophageal epithelium of adult mice which reveal that the stem cell gene, Sox2, is less essential for adult stem cell maintenance itself, but rather for stem cell proliferation and differentiation.</jats:p

p57Kip2 imposes the reserve stem cell state of gastric chief cells

Adult stem cells constantly react to local changes to ensure tissue homeostasis. In the main body of the stomach, chief cells produce digestive enzymes; however, upon injury, they undergo rapid proliferation for prompt tissue regeneration. Here, we identified p57Kip2 (p57) as a molecular switch for the reserve stem cell state of chief cells in mice. During homeostasis, p57 is constantly expressed in chief cells but rapidly diminishes after injury, followed by robust proliferation. Both single-cell RNA sequencing and dox-induced lineage tracing confirmed the sequential loss of p57 and activation of proliferation within the chief cell lineage. In corpus organoids, p57 overexpression induced a long-term reserve stem cell state, accompanied by altered niche requirements and a mature chief cell/secretory phenotype. Following the constitutive expression of p57 in vivo, chief cells showed an impaired injury response. Thus, p57 is a gatekeeper that imposes the reserve stem cell state of chief cells in homeostasis.11Nsciescopu