Biology subjects

Pollock, J.

Publications and source records attributed to Pollock, J..

10 recordsLinked to original sources

bioRxiv2026

Persistent activation of STAT6 in keratinocytes elicits neutrophilic skin inflammation, pruritus and S. aureus colonization reminiscent of chronic atopic dermatitis

Atopic dermatitis (AD) evolves from initial type 2 immunity-driven inflammation to chronic mixed responses by poorly understood mechanisms. To investigate how the prolonged activation of the usually IL-4/IL-13-induced transcription factor STAT6 in keratinocytes impacts on the development and subtype of AD, we generated a new mouse model in which a constitutively active form of STAT6 is selectively expressed in keratinocytes. These K14Cre+STAT6vt/vt mice spontaneously developed AD-like skin lesions characterized by Staphylococcus aureus colonization, neutrophilic inflammation, and pruritus starting at the age of 12-14 weeks. Treatment with antibiotics mitigated pathology, indicating that it is microbiota-driven. Comparison of human AD gene expression data with the transcriptome of skin biopsies from K14Cre+STAT6vt/vt mice revealed features shared with chronic AD, including genes associated with neutrophil and keratinocyte activation. Furthermore, heterozygous K14Cre+STAT6vt/wt mice developed a mixed eosinophilic and neutrophilic skin inflammation with exacerbated pathology compared to wild-type controls in an induced model of atopic dermatitis, compatible with chronic AD. These results indicate that persistent STAT6 activity in keratinocytes facilitates S. aureus outgrowth on the skin, promotes a type 1-/type 3-biased immune response, and is sufficient to mimic the transition from acute type 2 immunity-to chronic type 1-/type 3-immunity-dominated AD.

immunology↗

bioRxiv2025

ILC2s govern imprinting of alveolar macrophage-mediated immunity upon secondary helminth infection in the lung

Hookworm parasites migrate through the pulmonary system as part of their lifecycle, causing significant tissue damage that activates tissue-resident cell populations such as ILC2s. These cells drive type 2 immune responses critical for wound healing and the development of protective immunity. Macrophages, particularly monocyte-derived macrophages that seed the lung after infection, are central effectors of these responses and may be distinct from the tissue-resident populations they replace. However, the cellular and molecular factors governing the attrition of tissue-resident (TR) macrophages and their replacement remain poorly understood. We find that ILC2s and IL-4/IL-13-producing CD4+ T cells bi-directionally interact to regulate alveolar macrophage (AM) populations during infection. In the absence of these cells, type 2 immune responses are muted, leading to impaired anti-parasitic immunity in the lung during re-infection. This correlates with diminished expansion of type 2-polarized monocyte-derived alveolar macrophages (Mo-AMs), which are metabolically and transcriptionally distinct from TR-AMs. While Mo-AMs highly upregulated expression of Arg1, this was not essential for development of protective immunity in the lung. Concomitant with a more glycolytic metabolism, ILC2-induced Mo-AMs selectively upregulate arachidonate 15-lipoxygenase (Alox15), which promotes the capacity of macrophages to attack parasites in vitro. These results identify ILC2s as critical regulators of alveolar macrophage dynamics and important mediators of their function during helminth infection of the lung.

immunology↗

bioRxiv2025

Complexoform-restricted covalent TRMT112 ligands that allosterically agonize METTL5

Adaptors serve as hubs to regulate diverse protein complexes in cells. This multitude of functions can complicate the study of adaptors, as their genetic disruption may simultaneously impair the activities of several compositionally distinct complexes (or adaptor complexoforms). Here we describe the chemical proteomic discovery of bicyclopyrrolidine acrylamide stereoprobes that react with cysteine-100 (C100) of the methyltransferase (MT) adaptor TRMT112 in human cells. Curiously, the stereoprobes showed negligible reactivity with uncomplexed recombinant TRMT112, and we found that this interaction was restored excluively in the presence of METTL5, but not other MTs. A co-crystal structure revealed stereoprobe binding to a composite pocket proximal to C100 of TRMT112 that is templated by METTL5 and absent in other TRMT112:MT complexes. Structural rearrangements promoted by stereoprobe binding in turn lead to allosteric agonism of METTL5, thus revealing how covalent ligands targeting a pleiotropic adaptor can confer partner-specific functional effects through reactivity with a single complexoform.

biochemistry↗

bioRxiv2024

Covalent inhibitors of the RAS binding domain of PI3Ka impair tumor growth driven by RAS and HER2

Genetic disruption of the RAS binding domain (RBD) of PI 3-kinase (PI3K) prevents the growth of mutant RAS driven tumors in mice and does not impact PI3Ks role in insulin mediated control of glucose homeostasis. Selectively blocking the RAS-PI3K interaction may represent an attractive strategy for treating RAS-dependent cancers as it would avoid the toxicity associated with inhibitors of PI3K lipid kinase activity such as alpelisib. Here we report compounds that bind covalently to cysteine 242 in the RBD of PI3K p110 and block the ability of RAS to activate PI3K activity. These inhibitors have a profound impact on the growth of RAS mutant and also HER2 over-expressing tumors, particularly when combined with other inhibitors of the RAS/MAPK pathway, without causing hyperglycemia.

cancer biology↗

bioRxiv2024

AZIN2-dependent polyamine metabolism determines adipocyte progenitor fate and protects against obesity and dysmetabolism

Adipose tissue homeostasis plays a critical role in metabolic disease but the metabolic circuitry regulating adipose tissue dynamics remains unclear. In this study, polyamine metabolism emerges as an important regulator of adipose tissue pathophysiology. We identify AZIN2 (Antizyme inhibitor 2), a protein promoting polyamine synthesis and acetylation, as a major regulator of total acetyl-CoA in adipocyte progenitors (APs). AZIN2 deficient APs demonstrate increased H3K27 acetylation marks in genes related to lipid metabolism, cell cycle arrest and cellular senescence, and enhanced adipogenesis compared to wild-type counterparts. Upon high-fat diet (HFD)-induced obesity, global AZIN2 deficiency in mice provokes adipose tissue hypertrophy, AP senescence, lipid storage perturbations, inflammation and insulin resistance. IL4 promotes Azin2 expression in APs but not mature adipocytes due to diminished IL4 receptor expression in the latter. In human visceral and subcutaneous adipose tissue, AZIN2 expression positively correlates with expression of early progenitor markers and genes associated with protection against insulin resistance, while it negatively correlates with markers of lipogenesis. In sum, AZIN2-driven polyamine metabolism preserves adipose tissue health, a finding that could be therapeutically harnessed for the management of obesity-associated metabolic disease.

cell biology↗

bioRxiv2024

Suppression of NRF2-dependent cancer growth by a covalent allosteric molecular glue

The NRF2 transcription factor is constitutively active in cancer where it functions to maintain oxidative homeostasis and reprogram cellular metabolism. NRF2-active tumors exhibit NRF2-dependency and resistance to chemo/radiotherapy. Here we characterize VVD-065, a first-in-class NRF2 inhibitor that acts via an unprecedented allosteric molecular glue mechanism. In the absence of stress or mutation, NRF2 is rapidly degraded by the KEAP1-CUL3 ubiquitin-ligase complex. VVD-065 specifically and covalently engages C151 on KEAP1, which in turn promotes KEAP1-CUL3 complex formation, leading to enhancement of NRF2 degradation. Previously reported C151-directed compounds decrease KEAP1-CUL3 interactions and stabilize NRF2, thus establishing KEAP1_C151 as a tunable regulator of the KEAP1-CUL3 complex and NRF2 stability. VVD-065 inhibited NRF2-dependent tumor growth and sensitized cancers to chemo/radiotherapy, supporting an open Phase I clinical trial (NCT05954312).

cancer biology↗

bioRxiv2022

In situ quantification of osmotic pressure within living embryonic tissues

Mechanics is known to play a fundamental role in many cellular and developmental processes. Beyond active forces and material properties, osmotic pressure is believed to control essential cell and tissue characteristics. However, it remains very challenging to perform in situ and in vivo measurements of osmotic pressure. Here we introduce doubleemulsion droplet sensors that enable local measurements of osmotic pressure intra- and extra-cellularly within 3D multicellular systems, including living tissues. After generating and calibrating the sensors, we measured the osmotic pressure in blastomeres of early zebrafish embryos as well as in the interstitial fluid between the cells of the blastula by monitoring the size of droplets previously inserted in the embryo. Our results show a balance between intracellular and interstitial osmotic pressures, with values of approximately 0.7 MPa, but a large pressure imbalance between the inside and outside of the embryo. The ability to measure osmotic pressure in 3D multicellular systems (developing embryos, organoids, etc.) will help understand its role in fundamental biological processes.

biophysics↗

bioRxiv2020

Microbial Stimulation Reverses the Age-Related Decline in M Cells in Aged Mice

Ageing has a profound effect on the immune system, termed immunosenescence, resulting in increased incidence and severity of infections and decreased efficacy of vaccinations. We previously showed that immunosurveillance in the intestine, achieved primarily through antigen sampling M cells in the follicle associated epithelium (FAE) of Peyers patches, was compromised during ageing due to a decline in M cell functional maturation. The intestinal microbiota also changes significantly with age, but whether this affects M cell maturation was not known. We show that housing of aged mice on used bedding from young mice, or treatment with bacterial flagellin, were each sufficient to enhance the functional maturation of M cells in Peyers patches. An understanding of the mechanisms underlying the influence of the intestinal microbiota on M cells has the potential to lead to new methods to enhance the efficacy of oral vaccination in aged individuals.

immunology↗

bioRxiv2019

Resistance to change? The impact of group medication on AMR gene dynamics during commercial pig production

The anthropogenic selection of antimicrobial resistance (AMR) genes is under intense scrutiny, particularly in livestock production, where group antimicrobial administration is used to control disease. Whilst large epidemiological studies provide important data on the diversity and distribution of AMR genes, we have little insight into how group antimicrobial administration impacts AMR gene abundance and diversity within a system. Here, faecal microbiome and AMR gene dynamics were tracked for six months through a standard production cycle on a commercial pig unit. Our results demonstrate that specific AMR genes have reached an equilibrium across this farming system to the extent that the levels measured were maintained from suckling through to slaughter, despite increases in microbiome diversity in early development. These levels were not influenced by antibiotic use, either during the production cycle or following whole herd medication. Some AMR genes were found at levels higher than that of the bacterial 16S rRNA gene, indicating widespread distribution across the most common bacterial genera. The targeted AMR genes were also detected in nearby soil samples, several with decreasing abundance with increasing distance from the unit, demonstrating that the farm acts as a point source of AMR gene pollution. Metagenomic sequencing of a subset of samples identified 144 AMR genes, with higher gene diversity in the piglet samples compared to the sow samples. Critically, despite overwhelming and stable levels of resistance alleles against the main antimicrobials used on this farm, these compounds continue to control the bacterial pathogens responsible for production losses and compromised welfare.\n\nImportanceGroup antibiotic treatment has been used for decades to control bacterial diseases that reduce the productivity and compromise the welfare of livestock. Recent increases in antibiotic resistant infections in humans has resulted in concerns that antibiotic use in livestock may contribute to the development of untreatable bacterial infections in humans. There is however little understanding as to how the genes that bacteria require to become resistant to antibiotics respond during and after group antibiotic treatment of livestock, particularly in systems where high levels of antibiotics have been used for a prolonged period of time. We show that in such a system, levels of specific antibiotic resistance genes are high irrespective of group antibiotic treatments, whilst dramatic reductions in antibiotic use also fail to reduce the levels of these genes. These findings have important implications for public policy relating to the use of antibiotic in livestock farming.

microbiology↗

bioRxiv2019

The McGill Face Database: validation and insights into the recognition of facial expressions of complex mental states

Current databases of facial expressions of mental states typically represent only a small subset of expressions, usually covering the basic emotions (fear, disgust, surprise, happiness, sadness, and anger). To overcome these limitations, we introduce a new database of pictures of facial expressions reflecting the richness of mental states. 93 expressions of mental states were interpreted by two professional actors and high-quality pictures were taken under controlled conditions in front and side view. The database was validated with two different experiments (N=65). First, a four-alternative forced choice paradigm was employed to test the ability of participants to correctly select a term associated with each expression. In a second experiment, we employed a paradigm that did not rely on any semantic information. The task was to locate each face within a two-dimensional space of valence and arousal (mental state - space) employing a \"point-and-click\" paradigm. Results from both experiments demonstrate that subjects can reliably recognize a great diversity of emotional states from facial expressions. Interestingly, while subjects performance was better for front view images, the advantage over the side view was not dramatic. To our knowledge, this is the first demonstration of the high degree of accuracy human viewers exhibit when identifying complex mental states from only partially visible facial features. The McGill Face Database provides a wide range of facial expressions that can be linked to mental state terms and can be accurately characterized in terms of arousal and valence.

animal behavior and cognition↗