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Hay, R.

Publications and source records attributed to Hay, R..

4 recordsLinked to original sources

Cell-specific heteroclinic orbits govern interaction dynamics in inertial microfluidics for circulating tumour cell separation

Cancer survival rates increase with earlier diagnosis. However, many cancers are diagnosed only after symptoms develop, in the later stages of cancer progression. We need a diagnostic tool that can detect cancer earlier. Circulating tumour cells (CTCs) are cells that detach from the main tumour and can enter the bloodstream. The capture and analysis of CTCs can provide an early indicator of cancer. However, rarity of CTCs in blood makes their efficient capture challenging. Inertial microfluidics can be utilised for a label-free separation of CTCs from white blood cells (WBCs) by manipulating cells in the microchannel based on cell properties, achieving good separation performance. Inertial microfluidic devices mainly rely on size-based separation. However, residual WBC carryover limits complete separation. Here, we show that cell mechanical heterogeneity plays a large role in the separation process and should be considered in the design of these devices. We simulate the migration dynamics of CTCs and WBCs in a straight microchannel using a 3D lattice-Boltzmann-immersed-boundary-finite-element solver. Our results demonstrate that WBC migration behaviour changes depending on the deformability of a CTC. The size and deformability of the cells have been shown to determine the cell-specific heteroclinic orbits leading to different interaction types, these interactions alter WBC migration resulting in more/less WBC carryover. This work highlights that both cell-specific heteroclinic orbits and single cell migration rates should be considered in the design of inertial microfluidics for separation.

bioengineering↗

Numerical study of the size-based, shear-induced separation ofcirculating tumour cells from white blood cells in liquid biopsies

Circulating tumour cells (CTCs) are promising biomarkers for early cancer detection, yet their extreme rarity in blood necessitates efficient separation from white blood cells (WBCs) in lysed liquid biopsies. Inertial microfluidics offers a high-throughput, label-free approach to this challenge by leveraging size-dependent lateral migration. However, experimental observations reveal that WBCs migrate more rapidly than predicted, reducing separation performance. Using 3D lattice-Boltzmann-immersed-boundary-finite-element simulations, we characterized the migration dynamics of CTCs and WBCs in a straight microchannel. Our results reveal that the presence of a CTC enhances WBC cross-streamline migration, providing a mechanistic explanation for WBC contamination in CTC-enriched outlets. The numerical model capturing heterochiral orbital dynamics was validated experimentally, confirming the role of intercellular hydrodynamic interactions. These findings underscore the critical role of intercellular interactions in inertial microfluidic systems and provide guidance for optimizing suspension concentration and channel geometry to improve purity in rare cell isolation.

bioengineering↗

Genomic epidemiology of emerging terbinafine-resistant Trichophyton indotineae

Dermatophyte skin infections affect around a quarter of the worlds population and are a growing public health concern due to increasing incidence of novel species causing severe infections that are resistant to antifungal treatments. Trichophyton species cause the greatest burden of dermatophytosis worldwide, with the T. mentagrophytes species complex being particularly associated with the emergence of new aggressive infections. One emerging species, T. indotineae (originally T. mentagrophytes genotype VIII) is notable for the extensive nature of the often inflammatory infection, its clinical resistance to terbinafine antifungal treatment, and its rapid global spread. To better understand the epidemiology of this disease, we sourced isolates from severe cases of dermatophytosis in the United Kingdom, Ireland, France, Canada and India for the period 2018-2023, including the type strain from Japan. We used whole-genome sequencing to confirm 90 isolates were T. indotineae, and antifungal susceptibility testing indicated that over half of these (62%) were resistant to terbinafine (MIC [≥]1 mg/L). Pairwise genetic distances showed very high identity with only 147 (1-414) SNPs separating isolates that were nested within a monophyletic phylogeny, supporting a single evolutionary origin of T. indotineae. That no clear geographic clustering of isolates was observed confirms the rapid transcontinental spread of T. indotineae from its likely centre of diversity in Asia. Genome-wide analyses identified multiple non-synonymous SNPs in SQLE (ERG1), the squalene epoxidase target of terbinafine, that were associated with terbinafine in vitro resistance [≥]1 mg/L. However, five isolates exhibited high MIC values without SQLE mutations, suggesting the presence of alternative resistance mechanisms. Our findings highlight the importance of better genomic surveillance to understand and manage this severe and rapidly emerging terbinafine-resistant dermatophyte.

genomics↗

The proteasomal deubiquitinating enzyme PSMD14 regulates macroautophagy by controlling Golgi-to-ER retrograde transport

Ubiquitination regulates several biological processes. Here, we search for ubiquitin-related genes implicated in protein membrane trafficking performing a High-Content siRNA Screening including 1,187 genes of the human "ubiquitinome" using Amyloid Precursor Protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a novel key regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing or pharmacological inhibition of PSMD14 caused a robust and rapid inhibition of Golgi-to-ER retrograde transport which leads to a potent blockage of macroautophagy by a mechanism associated with the retention of Atg9A and Rab1A at the Golgi apparatus. Because pharmacological inhibition of the proteolytic core of the 20S proteasome did not recapitulate these effects, we concluded that PSMD14, and their K-63-Ub chains, act as a crucial regulator factor for macroautophagy by controlling Golgi-to-ER retrograde transport.

cell biology↗