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Wilkinson, L.

Publications and source records attributed to Wilkinson, L..

12 recordsLinked to original sources

PARNET: A CLIP-SEQ-BASED FOUNDATION MODEL FOR RNA SEQUENCE REPRESENTATION LEARNING

RNA-binding proteins (RBPs) orchestrate a complex combinatorial regulatory "code" that governs RNA splicing, stability, localization, and translation. Learning the relationship between RNA sequences and these processes is a central challenge in genomics. Foundation models, notably RNA language models, have emerged as the dominant approach, learning general-purpose representations from unlabeled sequence at scale. While RNA language models have demonstrated impressive performance across a broad range of downstream tasks, they generally learn from sequence reconstruction objectives alone, lacking direct connections to the regulatory principles that govern RNA function. Here we introduce Parnet, an RNA foundation model trained directly and exclusively on experimental CLIP-seq data. Parnet is a multi-task foundation model trained end-to-end on 223 eCLIP-seq experiments spanning 150 RBPs to predict base-resolution RBP binding profiles directly from RNA sequence. This CLIP-seq pretraining strategy departs fundamentally from the masked-language-modeling paradigm, anchoring learned RNA representations directly in measured protein-RNA interactions rather than sequence statistics. Parnet substantially outperforms its single-task predecessor RBPNet in binding profile and motif recovery, generalizes to unseen cell types and iCLIP data, and recapitulates position-dependent splicing regulation. Frozen Parnet embeddings, without task-specific fine-tuning, match or exceed the performance of both task-specific tools, as well as larger self-supervised RNA and genomic language models across diverse downstream tasks, including RNA biotype classification, lncRNA chromatin localization, translational efficiency, splice-site recognition, intron retention, and non-coding variant effect prediction. Importantly, Parnet remains mechanistically interpretable, tracing predictions back to the specific RBPs and motifs that drive them. These results establish the RBP interactome as a compact, functionally sufficient, and interpretable basis for foundation model pretraining in RNA biology.

bioinformatics↗

Bispecific Antibody Architecture and TNFRSF Target Selection Determine CD8+ T Cell Differentiation and Anti-tumour Immunity

Tumour necrosis factor receptor superfamily (TNFRSF)-targeting bispecific antibodies (bsAb) enable tumour-localised T cell co-stimulation, yet how antibody architecture and receptor choice govern activity remains unclear. Here we define how bsAb format and TNFRSF target selection shape CD8+ T cell responses and anti-tumour immunity. Using B7-H3 as a tumour-associated antigen, we show that dual-bivalent 2x2 bsAb elicit maximal agonistic activity, whereas the corresponding monovalent 1x1 format is least active. A 2x1 format retaining TNFRSF bivalency but monovalent B7-H3 binding preserves substantial activity, identifying co-stimulatory receptor bivalency as a key determinant of efficacy. Across TNFRSF targets, 4-1BB drives the strongest cytotoxic CD8+ T cell differentiation and anti-tumour response. This superiority is conserved in human T cells and reproduced by cognate ligands, indicating that the observed functional hierarchy reflects receptor-intrinsic biology. Mechanistically, efficacy requires tumour-associated B7-H3 and T cell-dependent 4-1BB signalling. Together, these findings establish general principles linking antibody architecture and receptor biology to co-stimulatory bsAb efficacy and provide a framework for rational design leading to optimal therapeutics.

immunology↗

The pore-forming protein APOL7C ruptures Leishmania major-containing parasitophorous vacuoles in dendritic cells to promote IL-12 production and the induction of T helper type 1 immunity

Classical dendritic cells (cDCs) are required for the efficient priming of T helper type 1 (Th1) immunity against Leishmania major infections in mice. They do so by producing large quantities of IL-12 in response to L. major infection. IL-12 in turn supports the generation of protective CD4+ Th1 cells. However, the mechanism(s) by which L. major instigates robust IL-12 production by cDCs has not been addressed. Here we show that expression of a pore-forming protein called APOL7C is induced in cDCs by L. major infection. APOL7C is expressed in the cytosol of host cells and, remarkably, upon infection is recruited to L. major parasitophorous vacuoles (PVs) where it induces PV rupture. PV rupture leads to the exposure of L. major-derived pathogen-associated molecular patterns (PAMPs) to cytosolic pattern recognition receptors (PRRs). Cytosolic PRR signaling then drives IL-12 production by cDCs. Consequently, L. major challenge of APOL7C-deficient mice results in reduced numbers of IL-12 producing cDCs as well as a diminished Th1 response. In sum, our data indicate the presence of pore-forming proteins in the cytosol of cDCs that interfere with the intracellular niche of Leishmania to modulate immunity to these parasites.

cell biology↗

REDUCED CACNA1C EXPRESSION PRODUCES ANHEDONIC REACTIONS TO PALATABLE SUCROSE IN RATS: NO INTERACTIONS WITH JUVENILE OR ADULT STRESS

Genetic variation in CACNA1C, which encodes the alpha-1 subunit of Cav1.2 L-type voltage-gated calcium channels, is strongly linked to risk for psychiatric disorders including schizophrenia, bipolar disorder, and major depression. Here we investigated the impact of mutations of one copy of Cacna1c on rats hedonic responses to palatable sucrose (assessed using the analysis of consumption microstructure). In addition, we also investigated the effects of combining either juvenile or adult stress with the manipulation of Cacna1c. Across three experiments Cacna1c+/- rats displayed attenuated hedonic reactions to sucrose compared to WT littermate controls, despite the Cacna1c+/- rats retaining sensitivity to sucrose concentration in terms of the amount of consumption. Unexpectedly, juvenile stress enhanced rather than reduced hedonic reactions to sucrose, while adult stress did not have clear hedonic effects. The effects of Cacna1c manipulation did not interact with either juvenile or adult stress. The fact that Cacna1c+/- rats display a clear analogue of anhedonia - a reduction in the positive hedonic reactions normally elicited by highly palatable sucrose - a symptom observed transdiagnostically across psychiatric disorders linked to CACNA1C, suggests this model may play a valuable role in the translational fiinvestigation of anhedonia.

genetics↗

Reduced gene dosage of the psychiatric risk gene Cacna1c is associated with impairments in hypothalamic-pituitary-adrenal axis activity in rats

Common and rare variation in CACNA1C gene expression has been consistently associated with neuropsychiatric disorders such as schizophrenia, bipolar disorder, and major depression, however the underlying biological pathways that cause this association have yet to be fully determined. In this study, we present evidence that rats with a reduced gene dosage of Cacna1c have increased basal corticosterone levels in the periphery and reduced Nr3c1 gene expression in the hippocampus and hypothalamus. These results are consistent with an effect of Cacna1c dosage on hypothalamus-pituitary-adrenal (HPA) axis function. We also show that the reduction of Nr3c1 in the hippocampus may be caused by epigenetic modification of exon 17 of Nr3c1, including the reduced interaction with the histone modifying markers H3K4me3 and H3K27ac. Heterozygous Cacna1c rats additionally show increased anxiety behaviours. These results support an association of Cacna1c heterozygosity with the altered activity of the HPA axis and function in the resting state and this may be a predisposing mechanism that contributes to the increased risk of psychiatric disorders with stress.

neuroscience↗

The pore-forming apolipoprotein APOL7C drives phagosomal rupture and antigen cross-presentation by dendritic cells

Type I conventional dendritic cells (cDC1s) are essential for the generation of protective cytotoxic T lymphocyte (CTL) responses against many types of viruses and tumours. They do so by internalizing antigens from virally infected or tumour cells and presenting them to CD8+ T cells in a process known as cross-presentation (XP). Despite the obvious biological importance of XP, the molecular mechanism(s) driving this process remain unclear. Here, we show that a cDC-specific pore-forming protein called apolipoprotein 7C (APOL7C) is upregulated in response to innate immune stimuli and is recruited to phagosomes. Strikingly, the association of APOL7C with phagosomes leads to phagosomal rupture, which in turn allows for the escape of engulfed protein antigens to the cytosol where they can be processed via the endogenous major histocompatibility complex (MHC) class I antigen processing pathway. We show that APOL7C recruitment to phagosomes is voltage-dependent and occurs in response to NADPH oxidase-induced depolarization of the phagosomal membrane. Our data indicate the presence of dedicated pore-forming apolipoproteins that mediate the delivery of phagocytosed proteins to the cytosol of activated cDC1s to facilitate MHC class I presentation of exogenous antigen and to regulate adaptive immunity.

cell biology↗

Behavioural consequences of Setd1a haploinsufficiency in mice: evidence for heightened emotional reactivity and impaired sensorimotor gating

A number of studies implicate the loss of function (LoF) mutations affecting the histone methyl transferase SETD1A in the aetiology of a range of neurodevelopmental disorders including schizophrenia. Here, we examined the behavioural consequences of haploinsufficiency of Setd1a in a mouse model. We find evidence for changes in a number of phenotypes of relevance to schizophrenia, including increased anxiety-related behaviour, enhanced acoustic startle response, and decreased pre-pulse inhibition of acoustic startle. The sensorimotor gating deficits in Setd1a+/- mice could not be rescued by haloperidol or risperidone, suggesting that these antipsychotics are ineffective for ameliorating schizophrenia-relevant phenotypes in Setd1a+/- mice and point to deficits in neural systems other than the monoamine system. These phenotypes are emerging as key features of a number of other mouse models of rare neurodevelopmental disorders caused by LoF mutations in genes encoding epigenome modifiers suggesting they may act in a network to modulate brain development. Taken together these data strengthen the support for the use of Setd1a haploinsufficient mice as a model for the biological basis of schizophrenia, and point towards possible underpinning neural mechanisms.

neuroscience↗

Analysis of diffusion tensor imaging data from UK Biobank confirms dosage effect of 15q11.2 copy-number variation on white matter and shows association with cognition

BackgroundCopy-number variations at the 15q11.2 BP1-BP2 locus are present in 0.5 to 1.0% of the population, and the deletion is associated with a range of neurodevelopmental disorders. Previously, we showed a reciprocal effect of 15q11.2 copy-number variation on fractional anisotropy, with widespread increases in deletion carriers. We aim to replicate and expand these findings, using a larger sample of participants (n=30,930), higher resolution imaging, and examining the implications for cognitive performance. MethodsDiffusion tensor imaging measures from participants with no neurological/psychiatric diagnoses were obtained from the UK Biobank database. We compared 15q11.2 BP1-BP2 deletion (n=103) and duplication (n=119) carriers to a large cohort of control individuals with no neuropsychiatric copy-number variants (n=29,870). Additionally, we assessed how changes in white matter mediated the association between carrier status and cognitive performance. ResultsDeletion carriers showed increases in fractional anisotropy in the internal capsule and cingulum, and decreases in the posterior thalamic radiation, compared to both duplication carriers and controls (who had intermediate values). Deletion carriers had lower scores across cognitive tasks compared to controls, which were mildly influenced by white matter alterations. Reduced fractional anisotropy in the posterior thalamic radiation partially contributed to worse cognitive performance in deletion carriers. ConclusionsThis study, together with our previous findings, provides convergent evidence for a dosage-dependent effect of 15q11.2 BP1-BP2 on white matter microstructure. Additionally, changes in white matter were found to partially mediate cognitive ability in deletion carriers, providing a link between white matter changes in 15q11.2 BP1-BP2 carriers and cognitive function.

neuroscience↗

Snord67 promotes lymph node metastasis and regulates U6-mediated alternative splicing in breast cancer

Small nucleolar RNAs (snoRNAs) have long been considered "housekeeping genes", important for ribosomal biogenesis and protein synthesis. However, there is increasing evidence that this largely ignored class of non-coding RNAs (ncRNAs) also have wide-ranging, non-canonical functions in diseases, including cancer. SnoRNAs have been shown to have both oncogenic and tumor suppressor roles, yet whether snoRNAs regulate metastasis is unknown. Here we show that expression of certain snoRNAs are enriched in lymph node (LN) metastases in a micro-surgical, immune-competent mouse model of breast cancer. We identify the snoRNA Snord67 as a key regulator of LN metastasis. Knockout of Snord67 resulted in significantly decreased LN tumor growth and subsequent development of distant metastases. This was associated with loss of targeted 2-O-methylation on the small nuclear RNA U6, a component of the spliceosome. RNA sequencing revealed distinct alternative splicing patterns in Snord67 knockout cells. Using rapid autopsy breast cancer cases, we found that matched human primary tumor and LN metastases revealed similar alternatively spliced genes, including several that are known to contribute to cancer. These results demonstrate that Snord67 is critical for growth of LN metastases and subsequent spread to distant metastases, and suggest that snoRNA-guided modifications of the spliceosome represent a previously unappreciated, yet targetable pathway in cancer.

cancer biology↗

Complement C3 and C3aR differentially impact on learned fear and innate anxiety

Complement is a key component of the immune system with roles in inflammation and host-defence. Here we reveal novel functions of complement pathways impacting on emotional reactivity of potential relevance to the emerging links between complement and risk for psychiatric disorder. We used mouse models to assess the effects of manipulating components of the complement system on emotionality. Mice lacking the complement C3a Receptor (C3aR-/-) demonstrated a selective increase in unconditioned (innate) anxiety whilst mice deficient in the central complement component C3 (C3-/-) showed a selective increase in conditioned (learned) fear. The dissociable behavioural phenotypes were linked to different signalling mechanisms. Effects on innate anxiety were independent of C3a, the canonical ligand for C3aR, consistent with the existence of an alternative ligand mediating innate anxiety, whereas effects on learned fear were due to loss of iC3b/CR3 signalling. Our findings show that specific elements of the complement system and associated signalling pathways contribute differentially to heightened states of anxiety and fear commonly seen in psychopathology.

neuroscience↗

Cyfip1 haploinsufficiency is associated with white matter changes, myelin thinning, reduction of mature oligodendrocytes and behavioural inflexibility

The biological basis of the increased risk for psychiatric disorders due to the pathogenic 15q11.2 copy number deletion is unknown. Previous work has shown disturbances in white matter tracts in human carriers of the deletion at this locus. Using a novel rat model, we have recapitulated low dosage of the candidate risk gene CYFIP1 present within the 15q11.2 interval to reveal alterations in white matter microstructure. Using diffusion tensor imaging, we first showed extensive white matter changes in Cyfip1 mutants that were most pronounced in the corpus callosum and external capsule. Transmission electron microscopy showed that these changes were associated with thinning of the myelin sheath in the corpus callosum. Myelin thinning was independent of changes in axon number or diameter but was associated with a reduction in the number of mature oligodendrocytes. Finally, we demonstrated functional effects on cognitive phenotypes sensitive to both disruptions in myelin and callosal circuitry.

neuroscience↗

Impulsive choice in mice lacking paternal expression of Grb10 provides evidence for intra-genomic conflict in behavior

Imprinted genes are expressed from one parental allele only as a consequence of epigenetic events that take place in the mammalian germ line and are thought to have evolved through intra-genomic conflict between parental alleles. We demonstrate, for the first time, oppositional effects of imprinted genes on brain and behavior. Specifically, here we show that mice lacking paternal Grb10 make fewer impulsive choices, with no dissociable effects on a separate measure of impulsive action. Taken together with previous work showing that mice lacking maternal Nesp55 make more impulsive choices this suggests that impulsive choice behavior is a substrate for the action of genomic imprinting. Moreover, the contrasting effect of these two genes suggests impulsive choices are subject to intra-genomic conflict and that maternal and paternal interests pull this behavior in opposite directions. Finally, these data may also indicate that an imbalance in expression of imprinted genes contributes to pathological conditions such as gambling and drug addiction, where impulsive behavior becomes maladaptive.

genetics↗