Biology subjects

Sherwood, M.

Publications and source records attributed to Sherwood, M..

6 recordsLinked to original sources

bioRxiv2026

Multi-omics analysis reveals integrin α3-dependent mechanisms of Zika virus oncolytic activity in pediatric neural tumor

Oncolytic virotherapy is an innovative approach to cancer treatment that uses replication-competent viruses to selectively target and destroy cancer cells while leaving healthy tissues largely unaffected. Zika virus (ZIKV), a neurotropic orthoflavivirus, has recently gained attention as a potential oncolytic agent due to its ability to infect neural-derived cells and suppress tumor growth in preclinical models. Although existing studies have examined ZIKVs oncolytic effects, the mechanisms underlying these effects remain largely unexplored. Additionally, the roles of individual ZIKV proteins and their interactions with host factors remain incompletely understood. Here, we used RNA sequencing, affinity purification-mass spectrometry, and functional assays to uncover previously unidentified mechanisms underlying ZIKVs oncolytic activity in pediatric neural tumors. We found that the ZIKV non-structural proteins NS4A and NS5 exert oncolytic effects, reducing tumorsphere size. ZIKV-host protein-protein interaction networks were characterized and showed that integrin 3 (gene: ITGA3), a mediator of cell-matrix adhesion, interacts with ZIKV NS2B and NS4A. Integrin 3 was further shown to be involved in ZIKV- and NS4A-induced tumorsphere size reduction, while ITGA3 knockdown and ZIKV infection additively inhibited 3D invasion. These findings provide critical mechanistic insights that could inform the rational design of ZIKV-based virotherapies and highlight opportunities for combination treatment strategies.

cancer biology↗

bioRxiv2026

Lymphatic vessel dysfunction contributes to severe dengue pathogenesis

Dengue virus (DENV) infection is a major global health threat, affecting more than half of the worlds population. Severe dengue is a life-threatening condition characterised by systemic bleeding, vascular leakage, and interstitial fluid accumulation that can progress to hypovolaemic shock. Circulating DENV non-structural protein 1 (NS1) has long been implicated in driving vascular hyperpermeability through its disruptive effects on endothelial cell junctions and the glycocalyx. The lymphatic system, which runs alongside the vascular network, plays a critical role in resorbing and recirculating interstitial fluid and immune cells extravasated from blood vessels. Despite its importance in maintaining tissue fluid homeostasis, the impact of dengue disease on lymphatic vessels has not previously been explored. Here, we present the first evidence that DENV-2 NS1 induces marked hyperpermeability in lymphatic endothelial cells, as measured by transendothelial electrical resistance, and impairs lymphangiogenesis in vitro. These effects were not attributable to changes in cell viability, morphology, or metabolic activity, as assessed by live/dead and metabolic assays and image analysis. Instead, we observed a defect in lymphatic endothelial cell migration, measured by scratch assay, which may underlie the reduced lymphangiogenic potential. Bulk RNA-seq, immunocytochemistry, and advanced image analysis further demonstrated pronounced reorganisation of cell-cell junctions, the cytoskeleton, and focal adhesions. Notably, junctional proteins including VE-cadherin, ZO-1, and Claudin-5 were not downregulated but instead displayed disorganised distribution along the cell junctions or aberrant cytoplasmic localisation. These structural disruptions became even more pronounced under flow conditions produced using a microfluidic system. Together, these findings demonstrate for the first time that DENV-2 NS1 directly disrupts lymphatic endothelial cell function, leading to junctional disorganisation and hyperpermeability. Such impairment of lymphatic drainage may contribute to the pathophysiology of severe dengue. Author SummaryDengue is a rapidly expanding mosquito-borne disease that now affects many tropical and subtropical regions worldwide. Severe cases can lead to extensive fluid leakage from blood vessels, which causes tissue swelling and, in the most dangerous situations, shock. Although much research has focused on how dengue damages the blood vascular system, almost nothing is known about its impact on the lymphatic system, which is responsible for removing fluid from tissues and returning it to the bloodstream. Because both systems work together to maintain fluid balance, understanding how dengue affects lymphatic vessels is important for explaining why fluid accumulation becomes so severe in critical disease. In our study, we examined whether the viral protein NS1, which circulates during infection, directly affects the cells that line lymphatic vessels. We found that NS1 increases the permeability of these cells and reduces their ability to form new vessel structures. These effects were not caused by cell death but by disruptions in how the cells organise their junctions, internal scaffolding, and interactions with neighbouring cells. By showing that NS1 can directly impair lymphatic vessel function, our work identifies a previously overlooked mechanism that may contribute to fluid build-up in severe dengue and suggests new avenues for future therapeutic research.

microbiology↗

bioRxiv2024

Multiomics analysis reveals key immunogenic signatures induced by oncolytic Zika virus infection of paediatric brain tumour cells

Brain tumours disproportionately affect children and are the largest cause of paediatric cancer-related death. Despite decades of research, paediatric standard-of-care therapy still predominantly relies on surgery, radiotherapy, and systemic use of cytotoxic chemotherapeutic agents, all of which can result in debilitating acute and late effects. Novel therapies that engage the immune system, such as oncolytic viruses (OVs), hold great promise and are desperately needed. Zika virus (ZIKV) infects and destroys aggressive cells from paediatric medulloblastoma, atypical teratoid rhabdoid tumour (ATRT), diffuse midline glioma (DMG), ependymoma and neuroblastoma. Despite this, the molecular mechanisms underpinning this therapeutic response are grossly unknown. By profiling the transcriptome across a time-course, we comprehensively investigated the response of paediatric medulloblastoma and ATRT brain tumour cells to ZIKV infection at the transcriptome level for the first time. We observed conserved TNF signalling pathway and cytokine signalling-related signatures following ZIKV infection. We demonstrated that the canonical TNF-alpha signalling pathway is implicated in oncolysis by reducing the viability of ZIKV-infected brain tumour cells and is a likely contributor to the anti-tumoural immune response through TNF-alpha secretion. Our findings have highlighted TNF-alpha as a potential prognostic marker for oncolytic ZIKV virotherapy. Performing a 49-plex ELISA, we generated the most comprehensive ZIKV-infected cancer cell secretome to date. We demonstrated that ZIKV infection induces a clinically relevant and diverse pro-inflammatory brain tumour cell secretome, thus circumventing the need for transgene modification to boost efficacy. We assessed publicly available scRNA-Seq data to model how the ZIKV-induced secretome may (i) interact with medulloblastoma tumour microenvironment (TME) cells via paracrine signalling and (ii) polarise lymph node immune cells via endocrine signalling. Our modelling has provided significant insight into the cytokine response that orchestrates the diverse anti-tumoural immune response during oncolytic ZIKV infection of brain tumours. Our findings have significantly contributed to understanding the molecular mechanisms governing oncolytic ZIKV infection and will help pave the way towards ZIKV-based virotherapy.

cancer biology↗

bioRxiv2024

Identification of natural Zika virus peptides presented on the surface of paediatric brain tumour cells by HLA classI

Despite decades of research, survival from brain cancer has scarcely improved and is drastically lower than that of other cancers. Novel therapies, such as immunotherapy, hold great promise for treating brain tumours and are desperately needed. Zika virus (ZIKV) infects and kills aggressive cancer cells with stem-like properties (CSCs) from both paediatric and adult brain tumours. Whilst T cell recruitment into ZIKV-infected brain tumours is becoming well documented, the specific mechanisms through which they are activated are poorly understood. We address this by employing a combined LC-MS/MS global proteome and immunopeptidome approach to describe, for the first time, human leukocyte antigen (HLA) presentation of ZIKV peptides on the surface of infected brain tumour cells. We first show that HLA class I (HLA-I) antigen processing & presentation is the most highly enriched immune response pathway in the global proteome of aggressive paediatric USP7-ATRT brain tumour cells following ZIKV infection. We identify USP7-ATRT cells as a good immunopeptidome model due to their homozygous of the globally most common HLA-A allotype (A*02:01). We predict the majority of the 19 ZIKV peptides that we identify here to strongly bind and be presented by HLA-A*02:01. We show that immunopeptide presentation corresponds with cellular ZIKV protein abundance, with ten peptides arising from the most abundant viral protein; non-structural protein 3 (NS3). Specifically, we show the ZIKV NS3 helicase domain to be a rich source of peptides. Finally, we verify that the identified ZIKV peptides do not mimic predicted peptides of the human proteome. The ZIKV peptides we identify here are potential candidates for developing novel epitope-specific brain tumour immunotherapies, and our findings provide potential insight into the efficacious cytotoxic T cell response that oncolytic ZIKV virotherapy can induce against brain tumours. Author SummaryViruses can attack cancer through two mechanisms: 1) infecting and killing the cancer cell and 2) activating the immune system against the tumour. Zika virus (ZIKV) uses both mechanisms to fight brain cancer. Here, we employ a powerful proteomic technique to identify fragments of viral proteins (peptides) presented by cell surface receptors on brain cancer cells infected with ZIKV. In the human body, immune system cells such as T cells recognise and become activated in response to these viral peptides and subsequently attack the infected patient tumour. We identify 19 ZIKV peptides, three of which have been shown previously to elicit T cell responses, four identified elsewhere, and twelve are novel. Our work helps delineate a component of how ZIKV acts as an immunotherapy, the T cell-specific immune response that the virus raises to promote clearance of brain tumours. The significance of our study is that the ZIKV peptides we identify may lead to the development of a novel brain tumour immunotherapy.

cancer biology↗

bioRxiv2022

Integrative transcriptomic and proteomic meta-analysis of Zika viral infection reveals potential mechanisms for oncolytic therapy in neuroblastoma

BACKGROUNDPaediatric neuroblastoma and brain tumours account for a third of all childhood cancer-related mortality. High-risk neuroblastoma is highly aggressive and survival is poor despite intensive multi-modal therapies with significant toxicity. Novel therapies are desperately needed. The Zika virus (ZIKV) is neurotropic and there is growing interest in employing ZIKV as a potential therapy against paediatric nervous system tumours, including neuroblastoma. METHODSHere, we perform an extensive meta-analysis of ZIKV infection studies to identify molecular mechanisms that may govern the oncolytic response in neuroblastoma cells. We summarise the neuroblastoma cell lines and ZIKV strains utilised and re-evaluate the infection data to deduce the susceptibility of neuroblastoma to the ZIKV oncolytic response. Integrating transcriptomics, interaction proteomics, dependency factor and compound datasets we show the involvement of multiple host systems during ZIKV infection. RESULTSWe identify that most paediatric neuroblastoma cell lines are highly susceptible to ZIKV infection and that the PRVABC59 ZIKV strain is the most promising candidate for neuroblastoma oncolytic virotherapy. ZIKV induces TNF signalling, lipid metabolism, the Unfolded Protein Response (UPR), and downregulates cell cycle and DNA replication processes. ZIKV is dependent on SREBP-regulated lipid metabolism and three protein complexes; V-ATPase, ER Membrane Protein Complex (EMC) and mammalian translocon. We propose ZIKV nonstructural protein 4B (NS4B) as a likely mediator of ZIKVs interaction with IRE1-mediated UPR, lipid metabolism and mammalian translocon. CONCLUSIONSOur work provides a significant understanding of ZIKV infection in neuroblastoma cells, which will facilitate the progression of ZIKV-based oncolytic virotherapy through pre-clinical research and clinical trials. KEYPOINTSO_LIThe Zika virus may provide the basis for an oncolytic virotherapy against Neuroblastoma C_LIO_LIMost paediatric neuroblastoma cell lines are susceptible to Zika viral infection C_LIO_LIWe identified molecular mechanisms that may induce the oncolytic response in Neuroblastoma C_LI Contribution to the fieldThe ability to both induce direct oncolysis and provoke an anti-tumoral immune response makes oncolytic virotherapy an attractive candidate to combat aggressive and heterogenous cancers, such as high-risk neuroblastoma. To progress oncolytic virotherapy to clinical trial it is essential to understand the host mechanisms the virus manipulates to kill cancer cells, alongside any pathology as a consequence of infection of normal cells. Here, we show that ZIKV efficiently infects and induces oncolysis of paediatric neuroblastoma cells and propose a potential TNF pathway-driven immune response. ZIKVs specificity for infection of nervous system cancer cells, while rarely causing nervous system-related pathology in young children, addresses many of its safety concerns. The inclusion of more effective and less toxic novel therapies, such as a potential ZIKV-based therapeutic, in multimodal treatment regimens will pave the way for improving patient long-term health and overall survival.

cancer biology↗

bioRxiv2021

Proteomic characterization of GSK3β knockout shows altered cell adhesion and metabolic pathway utilisation in colorectal cancer cells

Glycogen-specific kinase (GSK3{beta}) is an integral regulator of the Wnt signalling pathway as well as many other diverse signalling pathways and processes. Dys-regulation of GSK3{beta} is implicated in many different pathologies, including neurodegenerative disorders as well as many different tumour types. In the context of tumour development, GSK3{beta} has been shown to play both oncogenic and tumour suppressor roles, depending upon tissue, signalling environment or disease progression. Although multiple substrates of the GSK3{beta} kinase have been identified, the wider protein networks within which GSK3{beta} participates are not well known, and the consequences of these interactions not well understood. In this study, LC-MS/MS expression analysis was performed using knockout GSK3{beta} colorectal cancer cells and isogenic controls in colorectal cancer cell lines carrying dominant stabilizing mutations of {beta}-Catenin. Consistent with the role GSK3{beta}, we found that {beta}-Catenin levels and canonical Wnt activity are unaffected by knockout of GSK3{beta} and therefore use this knockout cell model to identify other processes in which GSK3{beta} is implicated. Quantitative proteomic analysis revealed perturbation of proteins involved in cell-cell adhesion, and we characterize the phenotype and altered proteomic profiles associated with this. We also characterize the perturbation of metabolic pathways resulting from GSK3{beta} knockout and identify defects in glycogen metabolism. In summary, using a precision colorectal cancer cell-line knockout model with constitutively activated {beta}-Catenin we are able to identify several of the diverse pathways and processes associated with GSK3{beta} function.

cancer biology↗