Biology subjects

Ford, H.

Publications and source records attributed to Ford, H..

4 recordsLinked to original sources

bioRxiv2026

Synergistic targeting of EP300/CBP and EYA co-activators collapses the rhabdomyosarcoma core regulatory circuit

Rhabdomyosarcoma (RMS) is a multi-subtype, high-risk pediatric sarcoma with a low mutational burden. The mutations found in RMS often alter genes involved in transcriptional control. Approaches to target dysregulated RMS transcription have remained elusive. Here, we develop a novel approach to target RMS transcription comprising simultaneous targeting of two distinctly acting transcriptional co-activators. We discover a common identity-controlling pan-RMS core regulatory circuit (CRC) composed of oncogenic and lineage-specific myogenic master transcription factors (mTFs). Using a super-enhancer-based reporter screen, we identify the EP300/CBP inhibitor A485 as a potent inhibitor of the pan-RMS CRC, though with efficacy-limiting toxicities. To enhance efficacy, we identify the mTF-binding co-activator EYA2 as a co-factor of this pan-RMS CRC and exploit a new second-generation EYA1/2 inhibitor, LG1-34, to disrupt its function. Combined co-activator inhibition inactivates the CRC and synergistically reduces RMS growth. This strategy dually targets CRC-associated co-activators to cooperatively suppress the RMS transcriptome and enforce cell death.

cancer biology↗

bioRxiv2023

Cryo-electron tomography of C. elegans mitochondria reveals how the ATP synthase dimer interface shapes crista membranes

Mitochondrial ATP synthases form rows of dimers, which induce membrane curvature to give cristae their characteristic lamellar or tubular morphology. The angle formed between the central stalks of ATP synthase dimers varies between species. Using cryo-electron tomography and sub-tomogram averaging, we determined the structure of the ATP synthase dimer from the nematode worm C. elegans and show that the dimer angle differs from previously determined structures. The consequences of this species-specific difference at the dimer interface were investigated by comparing C. elegans and S. cerevisiae mitochondrial morphology. We reveal that C. elegans has a larger ATP synthase dimer angle with more lamellar (flatter) cristae when compared to yeast. The underlying cause of this difference was investigated by generating an atomic model of the C. elegans ATP synthase dimer by homology modelling. A comparison of our C. elegans model to an existing S. cerevisiae structure reveals the presence of extensions and rearrangements in C. elegans subunits associated with maintaining the dimer interface. We speculate that increasing dimer angles could provide an advantage for species that inhabit variable-oxygen environments by forming flatter more energetically efficient cristae.

biochemistry↗

bioRxiv2022

The ExtRA Capacity Test: Reliability and validity study of a new tool for assessing shoulder muscle performance

ObjectivesThe primary objective was to evaluate the reliability of a new tool for assessing shoulder muscle performance: The ExtRA Capacity Test. The secondary objective was to assess whether this tool was a valid measure for assessing shoulder strength. MethodsThe ExtRA Capacity test involves two measures: maximal scapular plane lateral raises to 90{degrees} abduction with 2.5kg of external load and the maximal number of prone lying unsupported external rotations with the shoulder at 90{degrees} abduction. Both capacity tests are completed to a metronome set to 30 beats per minute 20 asymptomatic participants were sampled on 2 separate sessions, 1 week apart. The ExtRA Capacity Test was completed by the lead researcher and an independent physiotherapist. Shoulder strength was also measured using isokinetic dynamometry. ResultsThe test showed excellent inter-rater reliability (mean abduction ICC= 0.969, mean external rotation ICC= 0.822, with a 95% CI). Mean intra-rater variability was 3.96{+/-} 4.09 for the abduction measure and 1.70{+/-} 1.17 for the external rotation measure. Validity was calculated using Pearson correlation coefficient. The abduction measure showed good/ moderate correlation for the majority of strength measurements taken using isokinetic dynamometry however the external rotation capacity test did not correlate closely to isokinetic dynamometry strength measures. ConclusionThe abduction component of the ExtRA Capacity Test is a suitable measure for assessing shoulder strength in clinical practice. The external rotation measure is of suitable reliability however if used in clinical practice, it should not be used to assess shoulder strength, instead it may be suitable to assess movement control of the shoulder. SummaryVarious methods of measuring shoulder strength exist, ranging from cost free, relatively inaccurate methods to costly, complex methods which are of high reliability and validity but are challenging to use in a fast-pace, clinical environment. Objective outcome measures are used within a rehabilitation setting however at present there is no upper limb muscle performance test that is suitable to use on all people, regardless of strength or fitness level. Capacity testing of movements provides a functional, insight into strength specific to a real world/ sporting environment with lower limb capacity tests providing clear objective baselines that can be used for goal setting and providing return to play criteria following injury. This study proves the reliability and validity of the ExtRA Capacity Test which is a measure of shoulder muscle performance, suitable for people of all physical activity levels and upper limb strength.

physiology↗

bioRxiv2020

Variability in mitochondrial import, mitochondrial health and mtDNA copy number using Type II and Type V CRISPR effectors

Current methodologies for targeting the mitochondrial genome for basic research and/or therapeutic strategy development in mitochondrial diseases are restricted by practical limitations and technical inflexibility. The development of a functional molecular toolbox for CRISPR-mediated mitochondrial genome editing is therefore desirable, as this could enable precise targeting of mtDNA haplotypes using the precision and tuneability of CRISPR enzymes; however, published reports of "MitoCRISPR" systems have, to date, lacked reproducibility and independent corroboration. Here, we have explored the requirements for a functional MitoCRISPR system in human cells by engineering several versions of CRISPR nucleases, including the use of alternative mitochondrial protein targeting sequences and smaller paralogues, and the application of gRNA modifications that reportedly induce mitochondrial import. We demonstrate varied mitochondrial targeting efficiencies and influences on mitochondrial dynamics/function of different CRISPR nucleases, with Lachnospiraceae bacterium ND2006 (Lb) Cas12a being better targeted and tolerated than Cas9 variants. We also provide evidence of Cas9 gRNA association with mitochondria in HeLa cells and isolated yeast mitochondria, even in the absence of a targeting RNA aptamer. Finally, we present evidence linking mitochondrial-targeted LbCas12a/crRNA with increased mtDNA copy number dependent upon DNA binding and cleavage activity. We discuss reproducibility issues and the future steps necessary if MitoCRISPR is to be realised.

molecular biology↗