Biology subjects

Humphries, S.

Publications and source records attributed to Humphries, S..

10 recordsLinked to original sources

bioRxiv2025

DNA methylation reprogramming in marsupial embryos is restricted to the extraembryonic lineage

DNA methylation (5mC) is an epigenetic mark that plays a critical role in defining cell fate. Following fertilisation, DNA methylation inherited from gametes must be reprogrammed to establish totipotency and enable the parental-to-zygotic transition. To accomplish this, non-mammalian vertebrates such as zebrafish and medaka subtly reprogram maternal 5mC profiles while maintaining high methylation levels throughout embryogenesis. In contrast, eutherian mammals such as mouse and human undergo global 5mC erasure in both embryonic and extraembryonic lineages. However, while embryonic 5mC is rapidly re-established to high levels upon implantation, the trophectoderm, which gives rise to the placenta, displays sustained and conserved DNA hypomethylation, suggesting that this drastic 5mC erasure may be functionally linked to complex placentation in mammals. To clarify whether extensive post-fertilisation 5mC erasure co-evolved with placentation, we explored embryonic methylation dynamics in marsupials, a lineage of therian mammals with a short-lived placenta. We produced a near complete telomere-to-telomere (T2T) genome and generated detailed epigenome maps of embryonic development for an Australian marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata). We found the dunnart embryo exhibits genome wide DNA demethylation at the blastocyst stage, but these changes occur in the trophectoderm only, suggesting that 5mC erasure in the placenta is an ancestral state in therian mammals. Furthermore, the T2T-level dunnart genome assembly enabled identification of sex chromosomes, uncovering extensive hypomethylation of the paternally-inherited inactive X chromosome in females and revealing the previously unannotated master regulator of X chromosome inactivation, lncRNA Rsx. Our data indicate that while the use of genome-wide 5mC erasure differs between eutherian and marsupial lineages, 5mC erasure in extraembryonic tissue is ancestral to therian mammals and may be necessary to support placental development. HIGHLIGHTSO_LIFirst embryonic DNA methylation maps in an Australian marsupial C_LIO_LIExtensive global erasure of DNA methylation in the trophectoderm C_LIO_LIMaintenance of high DNA methylation in the embryonic lineage C_LIO_LIHypomethylated paternal X chromosome with methylated escapee genes C_LI

developmental biology↗

bioRxiv2024

Upregulated cholesterol biosynthesis facilitates the survival of methylation-retaining AML cells following decitabine treatment.

DNA hypomethylating agents (HMAs) are used to treat acute myeloid leukemia (AML) and myelodysplasia patients who are unsuitable for intensive chemotherapy. However, low response rates and therapy-resistant relapse remain significant challenges. To improve outcomes, we must understand how AML cells survive HMA treatment and continue to proliferate following therapy. We combine single-cell multiomics with parallel colony-forming assays to link HMA-induced heterogeneity with functional consequences in AML cellss. Azacytidine (AZA) and decitabine (DAC) induced global epigenetic heterogeneity, associated with upregulation of inflammatory responses and cell death pathways in a subset of hypomethylated cells. Some cells maintained high DNA methylation levels during treatment, and these methylation-retaining cells had increased self-renewal capacity following DAC treatment in two FLT3-ITD AML cell lines. Transcriptional profiling of colonies formed after HMA treatment revealed many genes with altered expression in both methylation-retaining and hypomethylated cells, with increased expression of cholesterol-related genes observed in all cell lines. Inhibition of the cholesterol biosynthesis pathway by rosuvastatin enhanced HMA effects on colony formation in vitro and extended survival in two in vivo models of AML. Our study demonstrates that HMA-induced epigenetic heterogeneity has implications for AML cell growth and identifies statins as a candidate co-treatment strategy to improve HMA efficacy.

cancer biology↗

bioRxiv2023

Hypoxia alters the effects of hypomethylating agents in acute myeloid leukaemia cells.

BackgroundAcute myeloid leukaemia (AML) is a deadly haematological malignancy that originates from mutated myeloid progenitor cells that lie quiescent in the hypoxic bone marrow. Elderly patients who cannot tolerate standard chemotherapies are administered low-dose hypomethylating agents (HMA) which act in a replication-dependent manner to reprogram the epigenome. Relapse is common following HMA treatment and may arise from quiescent leukaemia cells in the hypoxic bone marrow. Therefore, the effects of hypoxia on HMA efficacy may influence AML progression. ResultsAML cell lines (MOLM-13, MV-4-11, HL-60) were treated with decitabine (100nM) or azacitidine (500-2000nM) in normoxic (21% O2) and hypoxic (1% O2) conditions. Exposure to hypoxia significantly reduced AML cell growth across all cell lines, with no additional effects observed upon HMA treatment. This was associated with distinct effects on DNA methylation. The extent of hypomethylation induced by AZA treatment was reduced in hypoxia, whereas DAC-induced hypomethylation was maintained in low oxygen conditions. Transcriptional response to HMA treatment were also altered in hypoxia, with HMAs failing to up-regulate antigen presentation pathways in hypoxia. In particular, human leukocyte antigens (HLAs) such as HLA-DR were increased upon HMA treatment in normoxia, but not hypoxia. ConclusionOur results suggest that HMA-induced antigen presentation may be impaired in hypoxic tissues such as the bone marrow. This study highlights the need to consider microenvironmental factors when designing co-treatment strategies to improve HMA therapeutic efficacy.

cancer biology↗

bioRxiv2023

Optimal cell length for exploration and exploitation in chemotactic planktonic bacteria

Elongated morphologies are prevalent among motile bacterioplankton in aquatic systems. This is often attributed to enhanced chemotactic ability, but how long is best? We hypothesized the existence of an optimal cell length for efficient chemotaxis resulting from shape-imposed physical constraints acting on the trade-off between rapid exploration versus efficient exploitation of nutrient sources. To test this hypothesis, we first evaluated the chemotactic performance of elongated cephalexin-treated Escherichia coli towards -methyl-aspartate in an agarose-based microfluidic device that creates linear, stable and quiescent chemical gradients. Our experiments showed that cells of intermediate lengths aggregated most tightly to the chemoattractant source. We then replicated these experimental results with Individual Based Model (IBM) simulations. A sensitivity analysis of the IBM allowed us to gain mechanistic insights into which parameters drive this trend and showed that the poor chemotactic performance of very short cells is caused by loss of directionality, whereas long cells are penalized by brief, slow runs. Finally, we evaluated the chemotactic performance of cells of different length with IBM simulations of a phycosphere - a hotspot of microbial interactions in the ocean. Results indicated that long cells swimming in a run-and-reverse pattern with extended runs and moderate speeds are most efficient at harvesting nutrients in this microenvironment. The combination of microfluidic experiments and IBMs proves thus to be a powerful tool for untangling the physical constraints that motile bacteria are facing in aquatic systems.

ecology↗

bioRxiv2020

Phenotypic reconstruction of the last universal common ancestor reveals a complex cell

A fundamental concept in evolutionary theory is the last universal common ancestor (LUCA) from which all living organisms originated. While hypotheses regarding the phenotype of LUCA do exist, most are at best based on gene presence or absence. However, despite recent attempts to link genes and phenotypic traits in prokaryotes, it is still inherently difficult to predict phenotype based on the presence or absence of genes alone. Here, we apply a novel approach based on phylogenetically informed ancestral character reconstruction with 22 fundamental descriptors of prokaryotic cells to reconstruct LUCAs phenotype. We infer that the last universal common ancestor of all living organisms was likely an ovoid cell with a large genome, which probably had a cell wall and was actively motile. Potentially obtaining energy from the oxidation of inorganic compounds, tolerant of saltwater and thriving at temperatures above 70{degrees}C, LUCA was probably found living freely in anaerobic conditions in water of neutral pH. Our results depict LUCA as likely to be a far more complex cell than has previously been proposed, challenging the evolutionary model of increased complexity through time in prokaryotes. Given current estimates for the emergence of LUCA we suggest that early life very rapidly evolved considerable cellular complexity.

evolutionary biology↗

bioRxiv2020

Estimation of sinking velocities using free-falling dynamically scaled models: foraminifera as a test case

The velocity of settling particles is an important determinant of distribution in extinct and extant species with passive dispersal mechanisms, such as plants, corals, and phytoplankton. Here we adapt dynamic scaling, borrowed from engineering, to determine settling velocities. Dynamic scaling leverages physical models with relevant dimensionless numbers matched to achieve similar dynamics to the original object. Previous studies have used flumes, wind tunnels, or towed models to examine fluid flows around objects with known velocities. Our novel application uses free-falling models to determine the unknown sinking velocities of planktonic foraminifera - organisms important to our understanding of the Earths current and historic climate. Using enlarged 3D printed models of microscopic foraminifera tests, sunk in viscous mineral oil to match their Reynolds numbers and drag coefficients, we predict sinking velocities of real tests in seawater. This method can be applied to study other settling particles such as plankton, spores, or seeds. Summary StatementWe developed a novel method to determine the sinking velocities of biologically important microscale particles using 3D printed scale models.

biophysics↗

bioRxiv2020

Microrheology reveals microscale viscosity gradients in planktonic systems

Microbial activity in planktonic systems creates a dynamic and heterogeneous microscale seascape that harbours a diverse community of microorganisms and ecological interactions of global significance. In recent decades a great effort has been put into understanding this complex system, particularly focusing on the role of chemical patchiness, while overlooking a physical parameter that governs microbial life and is affected by biological activity: viscosity. Here we use microrheological techniques to measure viscosity at length scales relevant to microorganisms. Our results reveal the viscous nature and the spatial extent of the phycosphere, the microenvironment surrounding phytoplankton cells, and show heterogeneity in viscosity at the microscale. Such heterogeneity affects the distribution of chemicals and microorganisms, with pervasive and profound implications for the functioning of the planktonic ecosystem. One Sentence SummaryMicrorheology measurements unveil the existence of layers of increased viscosity surrounding phytoplankton cells and within aggregates.

ecology↗

bioRxiv2019

Metabolic dysregulation of the lysophospholipid/autotaxin axis in the chromosome 9p21 gene SNP rs10757274

AimsCommon chromosome 9p21 SNPs increase coronary heart disease (CHD) risk, independent of \"traditional lipid risk factors\". However, lipids comprise large numbers of structurally-related molecules not measured in traditional risk measurements, and many have inflammatory bioactivities. Here we applied lipidomic and genomic approaches to three model systems, to characterize lipid metabolic changes in common Chr9p21 SNPs which confer [~]30% elevated CHD risk associated with altered expression of ANRIL, a long ncRNA.\n\nMethods and ResultsUntargeted and targeted lipidomics was applied to plasma samples from Northwick Park Heart Study II (NPHSII) homozygotes for AA or GG in rs10757274. Elevated risk GG correlated with reduced lysophosphospholipids (lysoPLs), lysophosphatidic acids (lysoPA) and autotaxin (ATX). Five other risk SNPs did not show this phenotype. Correlation and network analysis showed that lysoPL-lysoPA interconversion was uncoupled from ATX in GG, indicating metabolic dysregulation. To identify candidate genes, transcriptomic data from shRNA downregulation of ANRIL in HEK293 cells was mined. Significantly-altered expression of several lysoPL/lysoPA metabolising enzymes was found (MBOAT2, PLA2G4C, LPCAT2, ACSL6, PNPLA2, PLBD1, PLPP1, PLPP2 and PLPPR2). Next, vascular smooth muscle cells differentiated from iPSCs of individuals homozygous for Chr9p21 risk SNPs were examined. Here, the presence of risk alleles was associated with altered expression of several lysoPL/lysoPA enzymes. Importantly, for several, deletion of the risk locus fully or partially reversed their expression to non-risk haplotype levels: ACSL3, DGKA, PLA2G2A, LPCAT2, LPL, PLA2G3, PNPLA3, PLA2G12A LIPC, LCAT, PLA2G6, ACSL1, MBOAT2.\n\nConclusionA Chr9p21 risk SNP associates with complex alterations in immune-bioactive phospholipids and their enzymatic metabolism. Lipid metabolites and genomic pathways associated with CHD pathogenesis in Chr9p21 and ANRIL-associated disease are demonstrated.\n\nOne sentence summaryInflammatory phospholipid metabolism defines a cardiovascular disease SNP

physiology↗

bioRxiv2018

Motile curved bacteria are Pareto-optimal

Curved-rods are a ubiquitous bacterial phenotype, but the fundamental question of why they are shaped this way remains unanswered. Through in silico experiments, we assessed freely swimming straight- and curved-rod bacteria of a wide diversity of equal-volume shapes parameterized by elongation and curvature, and predicted their performances in tasks likely to strongly influence overall fitness. Performance tradeoffs between these tasks lead to a variety of shapes that are Pareto-optimal, including coccoids, all straight rods, and a range of curvatures. Comparison with an extensive morphological survey of motile curved-rod bacteria indicates that the vast majority of species fall within the Pareto-optimal region of morphospace. This result is consistent with evolutionary tradeoffs between just three tasks: efficient swimming, chemotaxis, and low cell construction cost. We thus reveal the underlying selective pressures driving morphological diversity in a wide-spread component of microbial ecosystems.\n\nSignificance StatementBacteria exhibit a bewildering diversity of morphologies but despite their impact on nearly all aspects of life, they are frequently classified into a few general categories, usually just spheres and rods. Curved-rod bacteria are one simple variation and are widespread, particularly in the ocean. However, why so many species have evolved this shape is unknown. We show that curvature can increase swimming efficiency, revealing a widely-applicable selective advantage. Furthermore, we show that the distribution of cell lengths and curvatures observed across bacteria in nature are predicted by evolutionary tradeoffs between three tasks influenced by shape: efficient swimming, the ability to detect chemical gradients, and reduced cost of cell construction. We therefore reveal shape as an important component of microbial fitness.

biophysics↗

bioRxiv2017

150 million years of sustained increase in pterosaur flight efficiency

The long-term accumulation of biodiversity has been punctuated by remarkable evolutionary transitions that allowed organisms to exploit new ecological opportunities, and often resulted in large radiations of species. The Mesozoic flying reptiles - pterosaurs - which dominated the skies for over 150 million years (Myr), were the product of such a transition. The ancestors of pterosaurs were small and likely bipedal early archosaurs, which were certainly well adapted to terrestrial locomotion. More than 220 Myr ago, at some point in the Triassic, pterosaurs took flight and subsequently appear to have become capable and efficient flyers. However, the evolutionary processes that led to this efficiency remain enigmatic. Given the lack of proto-pterosaurs it is difficult to study how flight first evolved in this group, but we can test hypotheses about evolutionary changes to the energetics of locomotion following the transition to flight. Early pterosaurs were challenged by the c ...

evolutionary biology↗