ABSTRACT
In vitro tissue models hold great promise for modeling diseases and drug responses. Here, we used emulsion microfluidics to form micro-organospheres (MOSs), which are droplet-encapsulated miniature three-dimensional (3D) tissue models that can be established rapidly from patient tissues or cells. MOSs retain key biological features and responses to chemo-, targeted, and radiation therapies compared with organoids. The small size and large surface-to-volume ratio of MOSs enable various applications including quantitative assessment of nutrient dependence, pathogen-host interaction for anti-viral drug screening, and a rapid potency assay for chimeric antigen receptor (CAR)-T therapy. An automated MOS imaging pipeline combined with machine learning overcomes plating variation, distinguishes tumorspheres from stroma, differentiates cytostatic versus cytotoxic drug effects, and captures resistant clones and heterogeneity in drug response. This pipeline is capable of robust assessments of drug response at individual-tumorsphere resolution and provides a rapid and high-throughput therapeutic profiling platform for precision medicine.
Subject(s)
Antineoplastic Agents , Organoids , Antineoplastic Agents/pharmacology , Drug Evaluation, Preclinical/methods , Humans , Microfluidics , Precision MedicineABSTRACT
In Magnaporthe oryzae, pyriform conidia are the primary inoculum and the main source for disease dissemination in the field. In this study, we identified and characterized the COM1 gene that was disrupted in three insertional mutants producing slender conidia. COM1 encodes a putative transcription regulator unique to filamentous ascomycetes. The com1 disruption and deletion mutants had similar defects in conidium morphology and were significantly reduced in virulence on rice and barley seedlings. Microscopic examination revealed that the Deltacom1 mutants were defective in appressorium turgor generation, penetration, and infectious growth. COM1 was expressed constitutively in M. oryzae. The Com1 protein had putative helix-loop-helix structures and three predicted nuclear localization signal sequences. In transformants expressing COM1(335-613)-enhanced green fluorescent protein fusion constructs, fluorescence signals were observed in the nucleus. Our data indicated that the COM1 gene may encode a novel transcription regulator that regulates conidial development and invasive growth in M. oryzae.