Biology subjects

Clark, M. A.

Publications and source records attributed to Clark, M. A..

7 recordsLinked to original sources

bioRxiv2026

Metallothionein loss in cancer cells contributes to increased mutations through defective DNA repair and metabolic imbalance

Understanding which genes are involved in mutagenesis is essential for developing cancer prevention and treatment strategies; establishing protectors of the genome has revolutionized cancer biology. Here, we describe metallothionein (MT) proteins as previously uncharacterized protectors against mutagenesis. MT is a heavy metal binding protein essential for zinc homeostasis and protection against heavy metal cytotoxicity. Because zinc binds approximately 10-15% of the proteome and is critical for processes such as DNA repair and mitochondrial health, MT loss is expected to disrupt these processes. We hypothesized that MT loss induces genomic instability by impairing DNA repair and mitochondrial function. In this study, the consequences of MT deficiency in high-grade serous ovarian cancer (HGSC) were investigated by knockdown of the most highly expressed MT, MT2A. Loss of MT2A resulted in the impaired DNA repair pathway base excision repair (BER), leading to increased mutagenesis. MT2A deficiency produced mitochondrial dysfunction, characterized by a decrease in mitochondrial membrane potential, glycolysis, oxidative phosphorylation, amino acids, and an imbalance of nucleobases. Together, these defects reflect cellular states associated with increased cancer aggressiveness. These findings identify MT as a fundamental hub maintaining genomic and metabolic integrity. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=84 SRC="FIGDIR/small/736843v2_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@1af4adeorg.highwire.dtl.DTLVardef@15c588forg.highwire.dtl.DTLVardef@1ba5181org.highwire.dtl.DTLVardef@13d150b_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

bioRxiv2025

Group personality, rather than acoustic noise, causes variation in group decision-making in guppy shoals

Group living has essential fitness benefits for many species. While numerous studies have explored how environmental conditions impact collective movement, their impact on decisions made in a social context--a central component of group-living--is poorly documented. In this study, we assess how acoustic noise impacts group decision-making, cohesion and activity in fish shoals, using Trinidadian guppies (Poecilia reticulata) as a model species. Movements within a radially symmetric five-armed maze were measured using high-resolution trajectory data from video-tracking software. The behaviour of groups with and without continuous acoustic white noise were measured over a four-day testing period in a repeated-measures design. We found no significant change in swimming speed or group cohesion with additional acoustic noise. However, there was evidence for fewer following events (moves into already occupied arms) in the noise treatment compared to the control, but this additional noise had no effect on leadership attempts (moves into empty arms). We found strong evidence for consistent, repeatable differences between groups in all behavioural parameters indicating strong personality variation at the group level. Rather than environmental factors, these results provide evidence that consistent group-level differences dominate variation in collective behaviour, including group decision-making, in fish shoals.

animal behavior and cognition↗

bioRxiv2024

MYC is sufficient to generate mid-life high-grade serous ovarian and uterine serous carcinomas in a p53-R270H mouse model

Genetically engineered mouse models (GEMM) have fundamentally changed how ovarian cancer etiology, early detection, and treatment is understood. However, previous GEMMs of high-grade serous ovarian cancer (HGSOC) have had to utilize genetics rarely or never found in human HGSOC to yield ovarian cancer within the lifespan of a mouse. MYC, an oncogene, is amongst the most amplified genes in HGSOC, but it has not previously been utilized to drive HGSOC GEMMs. We coupled Myc and dominant negative mutant p53-R270H with a fallopian tube epithelium-specific promoter Ovgp1 to generate a new GEMM of HGSOC. Female mice developed lethal cancer at an average of 15.1 months. Histopathological examination of mice revealed HGSOC characteristics including nuclear p53 and nuclear MYC in clusters of cells within the fallopian tube epithelium and ovarian surface epithelium. Unexpectedly, nuclear p53 and MYC clustered cell expression was also identified in the uterine luminal epithelium, possibly from intraepithelial metastasis from the fallopian tube epithelium (FTE). Extracted tumor cells exhibited strong loss of heterozygosity at the p53 locus, leaving the mutant allele. Copy number alterations in these cancer cells were prevalent, disrupting a large fraction of genes. Transcriptome profiles most closely matched human HGSOC and serous endometrial cancer. Taken together, these results demonstrate the Myc and Trp53-R270H transgene was able to recapitulate many phenotypic hallmarks of HGSOC through the utilization of strictly human-mimetic genetic hallmarks of HGSOC. This new mouse model enables further exploration of ovarian cancer pathogenesis, particularly in the 50% of HGSOC which lack homology directed repair mutations. Histological and transcriptomic findings are consistent with the hypothesis that uterine serous cancer may originate from the fallopian tube epithelium.

cancer biology↗

bioRxiv2023

Anesthetic oxygen use and sex are critical factors in the FLASH sparing effect

IntroductionUltra-high dose-rate (UHDR) radiation has been reported to spare normal tissue compared to conventional dose-rate (CDR) radiation. However, reproducibility of the FLASH effect remains challenging due to varying dose ranges, radiation beam structure, and in-vivo endpoints. A better understanding of these inconsistencies may shed light on the mechanism of FLASH sparing. Here, we evaluate whether sex and/or use of 100% oxygen as carrier gas during irradiation contribute to the variability of the FLASH effect. MethodsC57BL/6 mice (24 male, 24 female) were anesthetized using isoflurane mixed with either room air or 100% oxygen. Subsequently, the mice received 27 Gy of either 9 MeV electron UHDR or CDR to a 1.6 cm2 diameter area of the right leg skin using the Mobetron linear accelerator. The primary post-radiation endpoint was time to full thickness skin ulceration. In a separate cohort of mice (4 male, 4 female) skin oxygenation was measured using PdG4 Oxyphor under identical anesthesia conditions. ResultsIn the UHDR group, time to ulceration was significantly shorter in mice that received 100% oxygen compared to room air, and amongst them female mice ulcerated sooner compared to males. However, no significant difference was observed between male and female UHDR mice that received room air. Oxygen measurements showed significantly higher tissue oxygenation using 100% oxygen as the anesthesia carrier gas compared to room air, and female mice showed higher levels of tissue oxygenation compared to males under 100% oxygen. ConclusionThe FLASH sparing effect is significantly reduced using oxygen during anesthesia compared to room air. The FLASH sparing was significantly lower in female mice compared to males. Both tissue oxygenation and sex are likely sources of variability in UHDR studies. These results suggest an oxygen-based mechanism for FLASH, as well as a key role for sex in the FLASH skin sparing effect.

cancer biology↗

bioRxiv2023

Discovery of a first-in-class small molecule ligand for WDR91 using DNA-encoded chemical library selection followed by machine learning

WD40 repeat-containing protein 91 regulates endosomal phosphatidylinositol 3-phosphate levels at the critical stage of endosome maturation and plays vital roles in endosome fusion, recycling, and transport by mediating protein-protein interactions. Due to its various roles in endocytic pathways, WDR91 has recently been identified as a potential host factor responsible for viral infection. We employed DNA-Encoded Chemical Library (DEL) selection against the WDR domain of WDR91, followed by machine learning to generate a model that was then used to predict ligands from the synthetically accessible Enamine REAL database. Screening of predicted compounds enabled us to identify the hit compound 1, which binds selectively to WDR91 with a KD of 6 {+/-} 2 M by surface plasmon resonance. The co-crystal structure confirmed the binding of 1 to the WDR91 side pocket, in proximity to cysteine 487. Machine learning-assisted structure activity relationship-by-catalog validated the chemotype of 1 and led to the discovery of covalent analogs 18 and 19. Intact mass LC-MS and differential scanning fluorimetry confirmed the formation of a covalent adduct, and thermal stabilization, respectively. The discovery of 1, 18, 19, accompanying SAR, and co-crystal structures will provide valuable insights for designing more potent and selective compounds against WDR91, thus accelerating the development of novel chemical tools to evaluate the therapeutic potential of WDR91 in disease.

molecular biology↗

bioRxiv2022

A novel method for investigating fish social behaviour in the field

Field studies of social behaviour are challenging due to having to observe multiple individuals simultaneously. In aquatic environments, these challenges are often amplified by environmental conditions such as habitat complexity, turbidity or darkness, which are often the conditions of interest in the context of studying anthropogenic impacts. Many fish species rely on social interactions for at least part of their life cycle, and while these interactions are known from laboratory studies to be impacted by environmental factors, approaches to quantify social behaviour under natural conditions are limited. Here, we present a novel method whereby multiple funnel traps are deployed simultaneously within a pond to quantify the aggregation and assortment of wild fish populations, using three-spine sticklebacks (Gasterosteus aculeatus) as a model species. The number of sticklebacks caught decreased in lower temperatures and through the seasons, from May to November. There was some evidence for decreases in fish numbers with reduced light intensity and higher dissolved oxygen levels. No effect of turbidity was found. The only evidence for changes in aggregation was related to the proportion of breeding males caught, where sticklebacks were less aggregated during the breeding season. These trends are expected based on previous work and knowledge about stickleback biology, validating the method. However, in contrast to previous studies, we found little evidence for phenotypic assortment. This study establishes a new cost-effective technique for investigating the social behaviour of wild fish, with some important benefits over existing techniques.

animal behavior and cognition↗

bioRxiv2018

Detecting HTS Barcode Contamination

DNA barcoding enables multiple samples to be characterized in parallel with high throughput sequencing (HTS) experiments for cost efficiencies. Cross-contamination of DNA barcode reagents can result in the detection of HTS sequences for barcodes that were not originally added to a particular sample. Cross-contamination of data between multiplexed samples can also occur. Avoidance and detection of contaminated barcodes is relevant for DNA forensic samples analysis, accurate cancer diagnosis, clinical research applications, metagenomic analysis, etc. We present recommendations for the avoidance of contamination and a tool, TallyBarcodes, to aid in the detection of DNA barcode contamination.

bioinformatics↗