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Biology subjects

Wilson, C. H.

Publications and source records attributed to Wilson, C. H..

6 recordsLinked to original sources

Combinatorial screening of biological degrader modules identifies heterobifunctional molecules that efficiently destroy MYC, leading to rapid tumour cell death in vitro and in vivo

Targeted protein degradation is a potent strategy against intracellular proteins impervious to traditional drugs. We describe a platform reliant on high-throughput cloning and mRNA-based delivery to quickly screen 100s-1000s of modular biological degraders, which we challenge against the high-turnover c-Myc oncoprotein. We uncover critical principles to drive degrader discovery against any target, free from tagging or prior cell line engineering, with direct applications in research and potentially therapy. We demonstrate the feasibility of targeted protein degradation of c-Myc across in vitro cancer models and in vivo xenograft model, uncovering distinct cellular responses between c-Myc degradation and inhibition.

bioengineering↗

A flexible, high-throughput system for studying mRNA translation kinetics in vitro and in cellulo with HiBit technology

HiBit is an engineered luciferases 11 amino acid component that can be introduced as a tag at either terminus of a protein of interest. When the LgBit component and a substrate are present, HiBit and LgBit dimerise forming a functional luciferase. The HiBit technology has been extensively used for high-throughput protein turnover studies in cells. Here, we have adapted the use of the HiBit technology to quantify mRNA translation temporally in vitro in the rabbit reticulocyte system and in cellulo in HEK293 cells constitutively expressing LgBit. The assay system can detect differences in Cap, 5UTR, modified nucleotide composition, coding sequence optimisation and poly(A) length. Importantly, using these assays we established the optimal mRNA composition varied depending on the encoded protein of interest, highlighting the importance of screening methods tailored to the protein of interest, and not reliant on reporter proteins. Our findings demonstrated that HiBit can be easily and readily adapted to monitor mRNA translation and offers a novel and highly favourable method for the development of mRNA-based therapeutics. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=149 SRC="FIGDIR/small/600987v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@92cec9org.highwire.dtl.DTLVardef@460f3dorg.highwire.dtl.DTLVardef@192a240org.highwire.dtl.DTLVardef@1da188e_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

HyperPRI: A Dataset of Hyperspectral Images for Underground Plant Root Study

Collecting and analyzing hyperspectral imagery (HSI) of plant roots over time can enhance our understanding of their function, responses to environmental factors, turnover, and relationship with the rhizosphere. Current belowground red-green-blue (RGB) root imaging studies infer such functions from physical properties like root length, volume, and surface area. HSI provides a more complete spectral perspective of plants by capturing a high-resolution spectral signature of plant parts, which have extended studies beyond physical properties to include physiological properties, chemical composition, and phytopathology. Understanding crop plants physical, physiological, and chemical properties enables researchers to determine high-yielding, drought-resilient genotypes that can withstand climate changes and sustain future population needs. However, most HSI plant studies use cameras positioned above ground, and thus, similar belowground advances are urgently needed. One reason for the sparsity of belowground HSI studies is that root features often have limited distinguishing reflectance intensities compared to surrounding soil, potentially rendering conventional image analysis methods ineffective. Here we present HyperPRI, a novel dataset containing RGB and HSI data for in situ, non-destructive, underground plant root analysis using ML tools. HyperPRI contains images of plant roots grown in rhizoboxes for two annual crop species - peanut (Arachis hypogaea) and sweet corn (Zea mays). Drought conditions are simulated once, and the boxes are imaged and weighed on select days across two months. Along with the images, we provide hand-labeled semantic masks and imaging environment metadata. Additionally, we present baselines for root segmentation on this dataset and draw comparisons between methods that focus on spatial, spectral, and spatialspectral features to predict the pixel-wise labels. Results demonstrate that combining HyperPRIs hyperspectral and spatial information improves semantic segmentation of target objects.

plant biology↗

A transient modified mRNA encoding Myc and Cyclin T1 induces cardiac regeneration and improves cardiac function after myocardial injury

Cardiac injury, such as myocardial infarction (MI), results in permanent loss of cardiomyocytes and in many cases heart failure. Transgenic expression of the pro-proliferative transcription factor Myc and Cyclin T1 can drive substantial adult cardiomyocyte proliferation to replace lost cardiomyocytes. Herein, we show that Myc and Cyclin T1 induced cardiomyocyte proliferation leads to myocardial repair and functional (long-term) recovery post-MI in mice. To provide a more translational approach, we developed modified mRNA (modRNA) encoding Myc-Ccnt1 as a transient and non-integrating strategy for regeneration. One dose of Myc-Ccnt1 modRNA is sufficient to transiently drives cardiomyocyte proliferation in human pluripotent stem cell-derived cardiomyocytes and a mouse MI model, where it leads to better heart function. Using single nuclei sequencing and proteomics, we show this was functionally mediated by transcriptional activation of cell-cycle regulating genes, which ultimately results in mitosis and cytokinesis of cardiomyocytes. Collectively, these findings indicate that Myc-Ccnt1 modRNA has the potential to be an effective regenerative therapeutic.

molecular biology↗

Transcriptomic profiling reveals a pro-nociceptive role for Angiotensin II in inflammatory bowel disease

ObjectiveVisceral pain is a leading cause of morbidity in inflammatory bowel disease (IBD), contributing significantly to reduced quality of life. Currently available analgesics often lack efficacy or have intolerable side-effects, driving the need for a more complete understanding of the mechanisms causing pain. MethodsWhole transcriptome gene expression analysis was performed by bulk RNA sequencing of colonic biopsies from patients with ulcerative colitis (UC) and Crohns disease (CD) reporting abdominal pain and compared with non-inflamed control biopsies. Putative pro-nociceptive mediators were identified based on pathway analysis of differentially expressed genes in IBD tissue and single cell gene expression in colonic neurons. Pro-nociceptive activity of identified mediators was assessed in assays of sensory neuron and colonic afferent activity. ResultsRNA sequencing analysis highlighted a 7.6-fold increase in the expression of angiotensinogen transcripts, Agt, the precursor to angiotensin II (Ang II), in samples from UC patients (p = 3.2x10-8). Consistent with the marked expression of the angiotensin AT1 receptor in colonic neurons, Ang II elicited an increase in intracellular Ca2+ in capsaicin-sensitive, voltage gated sodium channel subtype NaV1.8-positive sensory neurons. Ang II also evoked action potential discharge in high-threshold colonic nociceptors. These effects were inhibited by the AT1 receptor antagonist valsartan. ConclusionFindings from our study identify AT1 receptor-mediated colonic nociceptor activation as a novel pathway of visceral nociception in IBD patients with UC. This work highlights the potential utility of angiotensin receptor blockers, such as valsartan, as treatments for pain in IBD.

neuroscience↗

Insight into biogeochemical models from Scale Transition Theory: a dimensionless, scale-free approach

Leading an effective response to the accelerating crisis of anthropogenic climate change will require improved understanding of global carbon cycling. A critical source of uncertainty in Earth Systems Models (ESMs) is the role of microbes in mediating both the formation and decomposition of soil organic matter, and hence in determining patterns of CO2 efflux. Traditionally, ESMs model carbon turnover as a first order process impacted primarily by abiotic factors, whereas contemporary biogeochemical models often explicitly represent the microbial biomass and enzyme pools as the active agents of decomposition. However, the combination of non-linear microbial kinetics and ecological heterogeneity across space guarantees that upscaled dyamics will violate mean-field assumptions via Jensens Inequality. Violations of mean-field assumptions mean that parameter estimates from models fit to upscaled data (e.g. eddy covariance towers) are likely systematically biased. Here we present a generic mathematical analysis of upscaled michaelis-menten kinetics, grounded in Scale Transition Theory. We advance the framework by providing solutions in dimensionless form, and illustrate how this approach facilitates qualitative insight into the significance of this scale transition, and argue that it will facilitate future cross site intercomparisons of scale transition effects from flux data. We also discuss the critical terms that need to be constrained in order to unbias parameter estimates.

ecology↗