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Vaccine-induced immune thrombotic thrombocytopenia (VITT) has caused global concern. VITT is characterized by thrombosis and thrombocytopenia following COVID-19 vaccinations with the AstraZeneca ChAdOx1 nCov-19 and the Janssen Ad26.COV2.S vaccines. Patients present with thrombosis, severe thrombocytopenia developing 5 to 24 days following first dose of vaccine, with elevated D-dimer, and PF4 antibodies, signifying platelet activation. As of June 1, 2021, over 1.93 billion COVID-19 vaccine doses had been administered worldwide. Currently, 467 VITT cases (0.000024%) have been reported across the UK, Europe, Canada and Australia. Guidance on diagnosis and management of VITT has been reported but the pathogenic mechanism is yet to be fully elucidated. Here, we propose and discuss potential mechanisms in relation to adenovirus induction of VITT. We provide insights and clues into areas warranting investigation into the mechanistic basis of VITT, highlighting the unanswered questions. Further research is required to help solidify a pathogenic model for this condition.
Maha Othman; Alexander T. Baker; Elena Gupalo; Abdelrahman Elsebaie; Carly M. Bliss; Matthew T. Rondina; David Lillicrap; Alan L. Parker. To clot or not to clot? Ad is the question—Insights on mechanisms related to vaccine‐induced thrombotic thrombocytopenia. Journal of Thrombosis and Haemostasis 2021, 1 .
AMA StyleMaha Othman, Alexander T. Baker, Elena Gupalo, Abdelrahman Elsebaie, Carly M. Bliss, Matthew T. Rondina, David Lillicrap, Alan L. Parker. To clot or not to clot? Ad is the question—Insights on mechanisms related to vaccine‐induced thrombotic thrombocytopenia. Journal of Thrombosis and Haemostasis. 2021; ():1.
Chicago/Turabian StyleMaha Othman; Alexander T. Baker; Elena Gupalo; Abdelrahman Elsebaie; Carly M. Bliss; Matthew T. Rondina; David Lillicrap; Alan L. Parker. 2021. "To clot or not to clot? Ad is the question—Insights on mechanisms related to vaccine‐induced thrombotic thrombocytopenia." Journal of Thrombosis and Haemostasis , no. : 1.
The human adenovirus phylogenetic tree is split across seven species (A–G). Species D adenoviruses offer potential advantages for gene therapy applications, with low rates of pre-existing immunity detected across screened populations. However, many aspects of the basic virology of species D—such as their cellular tropism, receptor usage, and in vivo biodistribution profile—remain unknown. Here, we have characterized human adenovirus type 49 (HAdV-D49)—a relatively understudied species D member. We report that HAdV-D49 does not appear to use a single pathway to gain cell entry, but appears able to interact with various surface molecules for entry. As such, HAdV-D49 can transduce a broad range of cell types in vitro, with variable engagement of blood coagulation FX. Interestingly, when comparing in vivo biodistribution to adenovirus type 5, HAdV-D49 vectors show reduced liver targeting, whilst maintaining transduction of lung and spleen. Overall, this presents HAdV-D49 as a robust viral vector platform for ex vivo manipulation of human cells, and for in vivo applications where the therapeutic goal is to target the lung or gain access to immune cells in the spleen, whilst avoiding liver interactions, such as intravascular vaccine applications.
Emily Bates; John Counsell; Sophie Alizert; Alexander Baker; Natalie Suff; Ashley Boyle; Angela Bradshaw; Simon Waddington; Stuart Nicklin; Andrew Baker; Alan Parker. In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications. Viruses 2021, 13, 1483 .
AMA StyleEmily Bates, John Counsell, Sophie Alizert, Alexander Baker, Natalie Suff, Ashley Boyle, Angela Bradshaw, Simon Waddington, Stuart Nicklin, Andrew Baker, Alan Parker. In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications. Viruses. 2021; 13 (8):1483.
Chicago/Turabian StyleEmily Bates; John Counsell; Sophie Alizert; Alexander Baker; Natalie Suff; Ashley Boyle; Angela Bradshaw; Simon Waddington; Stuart Nicklin; Andrew Baker; Alan Parker. 2021. "In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications." Viruses 13, no. 8: 1483.
The human adenovirus phylogenetic tree is split across seven species (A-G). Species D adenoviruses offer potential advantages for gene therapy applications, with low rates of preexisting immunity detected across screened populations. However, many aspects of the basic virology of species D, such as their cellular tropism, receptor usage and in vivo biodistribution profile, remain unknown. Here, we have characterized human adenovirus type 49 (HAdV-D49), a relatively understudied species D member. We report that HAdV-D49 does not appear to use a single pathway to gain cell entry but appears able to interact with various surface molecules for entry. As such, HAdV-D49 can transduce a broad range of cell types in vitro, with variable engagement of blood coagulation FX. Interestingly, when comparing in vivo biodistribution to adenovirus type 5, HAdV-D49 vectors show reduced liver targeting whilst maintaining transduction of lung and spleen. Overall, this presents HAdV-D49 as a robust viral vector platform for ex vivo manipulation of human cells and for in vivo applications where the therapeutic goal is to target the lung or gain access to immune cells in the spleen whilst avoiding liver interactions, such as intravascular vaccine applications.
Emily A. Bates; John R. Counsell; Sophie Alizert; Alexander T. Baker; Natalie Suff; Ashley Boyle; Angela C. Bradshaw; Simon N. Waddington; Stuart A. Nicklin; Andrew H. Baker; Alan L. Parker. In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications. 2021, 1 .
AMA StyleEmily A. Bates, John R. Counsell, Sophie Alizert, Alexander T. Baker, Natalie Suff, Ashley Boyle, Angela C. Bradshaw, Simon N. Waddington, Stuart A. Nicklin, Andrew H. Baker, Alan L. Parker. In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications. . 2021; ():1.
Chicago/Turabian StyleEmily A. Bates; John R. Counsell; Sophie Alizert; Alexander T. Baker; Natalie Suff; Ashley Boyle; Angela C. Bradshaw; Simon N. Waddington; Stuart A. Nicklin; Andrew H. Baker; Alan L. Parker. 2021. "In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications." , no. : 1.
Adenovirus derived vectors, based on chimpanzee adenovirus Y25 (ChAdOx1) and human adenovirus type 26 are proving critical in combatting the 2019 SARS-CoV-2 pandemic. Following emergency use authorisation, scale up in vaccine administration has inevitably revealed vaccine related adverse effects; too rare to observe even in large Phase-III clinical trials. These include vaccine-induced thrombotic thrombocytopenia (VITT), an ultra-rare adverse event in which patients develop life-threatening blood clots 5-24 days following vaccination. To investigate vector-host interactions of ChAdOx1 underpinning VITT we solved the structure of the ChAdOx1 capsid by CryoEM, and the structure of the primary receptor tropism determining fiber-knob protein by crystallography. These structural insights have enabled us to unravel key protein interactions involved in ChAdOx1 cell entry and a possible means by which it may generate misplaced immunity to platelet factor 4 (PF4), a protein involved in coagulation. We use in vitro cell binding assays to show that the fiber-knob protein uses coxsackie and adenovirus receptor (CAR) as a high affinity binding partner, while it does not form a stable interface with CD46. Computational simulations identified a putative mechanism by which the ChAdOx1 capsid interacts with PF4 by binding in the spaces between hexon proteins, with downstream implications for the causes of VITT. Summary We present the structure of the ChAdOx1 viral vector, derived from chimpanzee adenovirus Y25 at 4.2Å resolution1. ChAdOx1 is in global use in the AstraZeneca vaccine, ChAdOx1 nCoV-19/AZD-1222, to combat the SARS-CoV-2 coronavirus pandemic. Recently observed, rare, adverse events make detailed mechanistic understanding of this vector key to informing proper treatment of affected patients and the development of safer viral vectors. Here, we determine a primary mechanism ChAdOx1 uses to attach to cells is coxsackie and adenovirus receptor (CAR), a protein which is identical in humans and chimpanzees. We demonstrate the vector does not form a stable CD46 interaction, a common species B adenovirus receptor, via its primary attachment protein. Further, we reveal the surface of the ChAdOx1 viral capsid has a strong electronegative potential. Molecular simulations suggest this charge, together with shape complementarity, are a mechanism by which an oppositely charged protein, platelet factor 4 (PF4) may bind the vector surface. PF4 is a key protein involved in the formation of blood clots2, and the target of auto-antibodies in heparin-induced immune thrombotic thrombocytopenia (HITT)3, an adverse reaction to heparin therapy which presents similarly to vaccine-induced immune thrombotic thrombocytopenia (VITT), a rare complication of ChAdOx1 nCoV-19 vaccination4–6. We propose a mechanism in which the ChAdOx1-PF4 complex may stimulate the production of antibodies against PF4, leading to delayed blood clot formation, as observed in VITT.
Alexander T. Baker; Ryan J. Boyd; Daipayan Sarkar; John Vant; Alicia Teijeira Crespo; Kasim Waraich; Chloe D. Truong; Emily Bates; Eric Wilson; Chun Kit Chan; Magdalena Lipka-Lloyd; Petra Fromme; Marius Bolni Nagalo; Meike Heurich; DeWight Williams; Po-Lin Chiu; Pierre J. Rizkallah; Alan L. Parker; Abhishek Singharoy; Mitesh J. Borad. The Structure of ChAdOx1/AZD-1222 Reveals Interactions with CAR and PF4 with Implications for Vaccine-induced Immune Thrombotic Thrombocytopenia. 2021, 1 .
AMA StyleAlexander T. Baker, Ryan J. Boyd, Daipayan Sarkar, John Vant, Alicia Teijeira Crespo, Kasim Waraich, Chloe D. Truong, Emily Bates, Eric Wilson, Chun Kit Chan, Magdalena Lipka-Lloyd, Petra Fromme, Marius Bolni Nagalo, Meike Heurich, DeWight Williams, Po-Lin Chiu, Pierre J. Rizkallah, Alan L. Parker, Abhishek Singharoy, Mitesh J. Borad. The Structure of ChAdOx1/AZD-1222 Reveals Interactions with CAR and PF4 with Implications for Vaccine-induced Immune Thrombotic Thrombocytopenia. . 2021; ():1.
Chicago/Turabian StyleAlexander T. Baker; Ryan J. Boyd; Daipayan Sarkar; John Vant; Alicia Teijeira Crespo; Kasim Waraich; Chloe D. Truong; Emily Bates; Eric Wilson; Chun Kit Chan; Magdalena Lipka-Lloyd; Petra Fromme; Marius Bolni Nagalo; Meike Heurich; DeWight Williams; Po-Lin Chiu; Pierre J. Rizkallah; Alan L. Parker; Abhishek Singharoy; Mitesh J. Borad. 2021. "The Structure of ChAdOx1/AZD-1222 Reveals Interactions with CAR and PF4 with Implications for Vaccine-induced Immune Thrombotic Thrombocytopenia." , no. : 1.
We previously developed a refined, tumor-selective adenovirus, Ad5NULL-A20, harboring tropism ablating mutations in each major capsid protein, to ablate all native means of infection. We incorporated a 20-mer peptide (A20) in the fiber knob for selective infection via αvβ6 integrin, a marker of aggressive epithelial cancers. Methods: To ascertain the selectivity of Ad5NULL-A20 for αvβ6-positive tumor cell lines of pancreatic and breast cancer origin, we performed reporter gene and cell viability assays. Biodistribution of viral vectors in mice harboring xenografts with low, medium, and high αvβ6 levels was quantified by qPCR for viral genomes 48 h post intravenous administration. Results: Ad5NULL-A20 vector transduced cells in an αvβ6-selective manner, whilst cell killing mediated by oncolytic Ad5NULL-A20 was αvβ6-selective. Biodistribution analysis following intravenous administration into mice bearing breast cancer xenografts demonstrated that Ad5NULL-A20 resulted in significantly reduced liver accumulation coupled with increased tumor accumulation compared to Ad5 in all three models, with tumor-to-liver ratios improved as a function of αvβ6 expression. Conclusions: Ad5NULL-A20-based virotherapies efficiently target αvβ6-integrin-positive tumors following intravenous administration, validating the potential of Ad5NULL-A20 for systemic applications, enabling tumor-selective overexpression of virally encoded therapeutic transgenes.
James Davies; Gareth Marlow; Hanni Uusi-Kerttula; Gillian Seaton; Luke Piggott; Luned Badder; Richard Clarkson; John Chester; Alan Parker. Efficient Intravenous Tumor Targeting Using the αvβ6 Integrin-Selective Precision Virotherapy Ad5NULL-A20. Viruses 2021, 13, 864 .
AMA StyleJames Davies, Gareth Marlow, Hanni Uusi-Kerttula, Gillian Seaton, Luke Piggott, Luned Badder, Richard Clarkson, John Chester, Alan Parker. Efficient Intravenous Tumor Targeting Using the αvβ6 Integrin-Selective Precision Virotherapy Ad5NULL-A20. Viruses. 2021; 13 (5):864.
Chicago/Turabian StyleJames Davies; Gareth Marlow; Hanni Uusi-Kerttula; Gillian Seaton; Luke Piggott; Luned Badder; Richard Clarkson; John Chester; Alan Parker. 2021. "Efficient Intravenous Tumor Targeting Using the αvβ6 Integrin-Selective Precision Virotherapy Ad5NULL-A20." Viruses 13, no. 5: 864.
Background: We previously developed a refined, tumor selective adenovirus, Ad5NULL-A20, har-boring tropism ablating mutations in each major capsid protein, to ablate all native means of infection. We incorporated a 20mer peptide (A20) in the fiber knob for selective infection via αvβ6 integrin, a marker of aggressive epithelial cancers. Methods: To ascertain the selectivity of Ad5NULL-A20 for αvβ6 positive tumor cell lines of pancreatic and breast cancer origin, we performed reporter gene and cell viability assays. Biodistribution of viral vectors in mice harboring xenografts with low, medium, and high αvβ6 levels was quantified by qPCR for viral genomes 48 hours post intravenous administration. Results: Ad5NULL-A20 vector transduced cells in an αvβ6 selective manner, whilst cell killing me-diated by oncolytic Ad5NULL-A20 was αvβ6 selective. Biodistribution analysis following intrave-nous administration into mice bearing breast cancer xenografts demonstrated that Ad5NULL-A20 resulted in significantly reduced liver accumulation coupled with increased tumor accumulation compared to Ad5 in all three models, with tumor: liver ratios improved as a function of αvβ6 expression. Conclusions: Ad5NULL-A20 based virotherapies efficiently target αvβ6 integrin positive tumors following intravenous administration, validating the potential of Ad5NULL-A20 for systemic ap-plications, enabling tumor selective overexpression of virally encoded therapeutic transgenes.
James A Davies; Gareth Marlow; Hanni K Uusi-Kerttula; Gillian Seaton; Luke Piggott; Richard W. E. Clarkson; John D Chester; Alan L. Parker. Efficient Intravenous Tumor Targeting Using the αVβ6 Integrin Selective Precision Virotherapy Ad5NULL-A20. 2021, 1 .
AMA StyleJames A Davies, Gareth Marlow, Hanni K Uusi-Kerttula, Gillian Seaton, Luke Piggott, Richard W. E. Clarkson, John D Chester, Alan L. Parker. Efficient Intravenous Tumor Targeting Using the αVβ6 Integrin Selective Precision Virotherapy Ad5NULL-A20. . 2021; ():1.
Chicago/Turabian StyleJames A Davies; Gareth Marlow; Hanni K Uusi-Kerttula; Gillian Seaton; Luke Piggott; Richard W. E. Clarkson; John D Chester; Alan L. Parker. 2021. "Efficient Intravenous Tumor Targeting Using the αVβ6 Integrin Selective Precision Virotherapy Ad5NULL-A20." , no. : 1.
Oncolytic viruses possess the ability to infect, replicate and lyse malignantly transformed tumour cells. This oncolytic activity amplifies the therapeutic advantage and induces a form of immunogenic cell death, characterised by increased CD8+ T‐cell infiltration into the tumour microenvironment. This important feature of oncolytic viruses can result in the warming up of immunologically “cold” tumour types, presenting the enticing possibility that oncolytic virus treatment combined with immunotherapies may enhance efficacy. In this review, we assess some of the most promising candidates that might be used for oncolytic virotherapy: immunotherapy combinations. We assess their potential as separate agents or as agents combined into a single therapy, where the immunotherapy is encoded within the genome of the oncolytic virus. The development of such advanced agents will require increasingly sophisticated model systems for their preclinical assessment and evaluation. In vivo rodent model systems are fraught with limitations in this regard. Oncolytic viruses replicate selectively within human cells, and therefore require human xenografts in immune‐deficient mice for their evaluation. However, the use of immune‐deficient rodent models hinders the ability to study immune responses against any immunomodulatory transgenes engineered within the viral genome and expressed within the tumour microenvironment. There has therefore been a shift towards the use of more sophisticated ex vivo, patient derived model systems based on organoids and explant co‐cultures with immune cells, which may be more predictive of efficacy than contrived and artificial animal models. We review the best of those model systems here.
Alicia Teijeira Crespo; Stephanie Burnell; Lorenzo Capitani; Rebecca Bayliss; Elise Moses; Georgina H. Mason; James A. Davies; Andrew J. Godkin; Awen M. Gallimore; Alan L. Parker. Pouring petrol on the flames: Using oncolytic virotherapies to enhance tumour immunogenicity. Immunology 2021, 163, 389 -398.
AMA StyleAlicia Teijeira Crespo, Stephanie Burnell, Lorenzo Capitani, Rebecca Bayliss, Elise Moses, Georgina H. Mason, James A. Davies, Andrew J. Godkin, Awen M. Gallimore, Alan L. Parker. Pouring petrol on the flames: Using oncolytic virotherapies to enhance tumour immunogenicity. Immunology. 2021; 163 (4):389-398.
Chicago/Turabian StyleAlicia Teijeira Crespo; Stephanie Burnell; Lorenzo Capitani; Rebecca Bayliss; Elise Moses; Georgina H. Mason; James A. Davies; Andrew J. Godkin; Awen M. Gallimore; Alan L. Parker. 2021. "Pouring petrol on the flames: Using oncolytic virotherapies to enhance tumour immunogenicity." Immunology 163, no. 4: 389-398.
Adenoviruses are powerful tools experimentally and clinically. To maximize efficacy, the development of serotypes with low preexisting levels of immunity in the population is desirable.
Alexander T. Baker; James A. Davies; Emily A. Bates; Elise Moses; Rosie M. Mundy; Gareth Marlow; David K. Cole; Carly M. Bliss; Pierre J. Rizkallah; Alan L. Parker. The Fiber Knob Protein of Human Adenovirus Type 49 Mediates Highly Efficient and Promiscuous Infection of Cancer Cell Lines Using a Novel Cell Entry Mechanism. Journal of Virology 2021, 95, 1 .
AMA StyleAlexander T. Baker, James A. Davies, Emily A. Bates, Elise Moses, Rosie M. Mundy, Gareth Marlow, David K. Cole, Carly M. Bliss, Pierre J. Rizkallah, Alan L. Parker. The Fiber Knob Protein of Human Adenovirus Type 49 Mediates Highly Efficient and Promiscuous Infection of Cancer Cell Lines Using a Novel Cell Entry Mechanism. Journal of Virology. 2021; 95 (4):1.
Chicago/Turabian StyleAlexander T. Baker; James A. Davies; Emily A. Bates; Elise Moses; Rosie M. Mundy; Gareth Marlow; David K. Cole; Carly M. Bliss; Pierre J. Rizkallah; Alan L. Parker. 2021. "The Fiber Knob Protein of Human Adenovirus Type 49 Mediates Highly Efficient and Promiscuous Infection of Cancer Cell Lines Using a Novel Cell Entry Mechanism." Journal of Virology 95, no. 4: 1.
More people are surviving longer with cancer. Whilst this can be partially attributed to advances in early detection of cancers, there is little doubt that the improvement in survival statistics is also due to the expansion in the spectrum of treatments available for efficacious treatment. Transformative amongst those are immunotherapies, which have proven effective agents for treating immunogenic forms of cancer, although immunologically “cold” tumour types remain refractive. Oncolytic viruses, such as those based on adenovirus, have great potential as anti-cancer agents and have seen a resurgence of interest in recent years. Amongst their many advantages is their ability to induce immunogenic cell death (ICD) of infected tumour cells, thus providing the alluring potential to synergise with immunotherapies by turning immunologically “cold” tumours “hot”. Additionally, enhanced immune mediated cell killing can be promoted through the local overexpression of immunological transgenes, encoded from within the engineered viral genome. To achieve this full potential requires the development of refined, tumour selective “precision virotherapies” that are extensively engineered to prevent off-target up take via native routes of infection and targeted to infect and replicate uniquely within malignantly transformed cells. Here, we review the latest advances towards this holy grail within the adenoviral field.
Tabitha G. Cunliffe; Emily A. Bates; Alan L. Parker. Hitting the Target but Missing the Point: Recent Progress towards Adenovirus-Based Precision Virotherapies. Cancers 2020, 12, 3327 .
AMA StyleTabitha G. Cunliffe, Emily A. Bates, Alan L. Parker. Hitting the Target but Missing the Point: Recent Progress towards Adenovirus-Based Precision Virotherapies. Cancers. 2020; 12 (11):3327.
Chicago/Turabian StyleTabitha G. Cunliffe; Emily A. Bates; Alan L. Parker. 2020. "Hitting the Target but Missing the Point: Recent Progress towards Adenovirus-Based Precision Virotherapies." Cancers 12, no. 11: 3327.
More people are surviving longer with cancer. Whilst this can be partially attributed to advances in early detection of cancers, there is little doubt that the improvement in survival statistics is also due to the expansion in the spectrum of treatments available for efficacious treatment. Transformative amongst those are immunotherapies, which have proven effective agents for treating immunogenic forms of cancer, though immunologically “cold” tumour types remain refractive. Oncolytic viruses, such as those based on adenovirus have great potential as anti-cancer agents and have seen a resurgence of interest in recent years. Amongst their many advantages is their ability to induce immunogenic cell death (ICD) of infected tumour cells, thus providing the alluring potential to synergize with immunotherapies by turning immunologically “cold” tumours “hot”. Additionally, enhanced immune mediated cell killing can be promoted through the local overexpression of immunological transgenes, encoded from within the engineered viral genome. To achieve this full potential requires the development of refined, tumour selective “precision virotherapies” that are extensively engineered to prevent off-target up take via native routes of infection, and targeted to infect and replicate uniquely within malignantly transformed cells. Here, we review the latest advances towards this holy grail within the adenoviral field.
Tabitha G. Cunliffe; Emily A. Bates; Alan L. Parker. Hitting the Target but Missing the Point: Recent Progress Towards Adenovirus-Based Precision Virotherapies. 2020, 1 .
AMA StyleTabitha G. Cunliffe, Emily A. Bates, Alan L. Parker. Hitting the Target but Missing the Point: Recent Progress Towards Adenovirus-Based Precision Virotherapies. . 2020; ():1.
Chicago/Turabian StyleTabitha G. Cunliffe; Emily A. Bates; Alan L. Parker. 2020. "Hitting the Target but Missing the Point: Recent Progress Towards Adenovirus-Based Precision Virotherapies." , no. : 1.
The human adenovirus (HAdV) phylogenetic tree is diverse, divided across seven species and comprising over 100 individual types. Species D HAdV are rarely isolated with low rates of pre-existing immunity, making them appealing for therapeutic applications. Several species D vectors have been developed as vaccines against infectious diseases where they induce robust immunity in pre-clinical models and early phase clinical trials. However, many aspects of the basic virology of species D HAdV, including their basic receptor usage and means of cell entry, remain understudied.Here, we investigated HAdV-D49, which previously has been studied for vaccine and vascular gene transfer applications. We generated a pseudotyped HAdV-C5 presenting the HAdV-D49 fiber knob protein (HAdV-C5/D49K). This pseudotyped vector was efficient at infecting cells devoid of all known HAdV receptors, indicating HAdV-D49 uses an unidentified cellular receptor. Conversely, a pseudotyped vector presenting the fiber knob protein of the closely related HAdV-D30 (HAdV-C5/D30K), differing in just four amino acids from HAdV-D49, failed to demonstrate the same tropism. These four amino acid changes resulted in a change in isoelectric point of the knob protein, with HAdV-D49K presenting a highly basic region around the apex compared to HAdV-D30K. Structurally and biologically we demonstrate that HAdV-D49 knob protein is unable to engage CD46, whilst potential interaction with CAR is extremely limited by extension of the DG loop. HAdV-C5/49K efficiently transduced cancer cell lines of pancreatic, breast, lung, oesophageal and ovarian origin, indicating it may have potential for oncolytic virotherapy applications, especially for difficult to transduce tumour types.ImportanceAdenoviruses are powerful tools experimentally and clinically. To maximise efficacy, the development of serotypes with low levels of pre-existing immunity in the population is desirable. Consequently, attention has focussed on those derived from species D, which have proven robust vaccine platforms. This widespread usage is despite limited knowledge underpinning their basic biology and cellular tropism.We investigated the tropism of HAdV-D49, demonstrating it uses a novel cell entry mechanism that bypasses all known HAdV receptors. We demonstrate, biologically, that a pseudotyped HAdV-C5/D49K vector efficiently transduces a wide range of cell lines, including those presenting no known adenovirus receptor. Structural investigation suggests that this broad tropism is the result of a highly basic surface electrostatic potential, since a homologous pseudotyped vector with a more acidic surface potential, HAdV-C5/D30K, does not have a similar pan-tropism. Therefore, HAdV-C5/D49K may form a powerful vector for therapeutic applications capable of infecting difficult to transduce cells.
Alexander T. Baker; Gareth L Marlow; James A. Davies; Elise Moses; Rosie M. Mundy; David K. Cole; Pierre J. Rizkallah; Alan L. Parker. The fiber knob protein of human adenovirus type 49 mediates highly efficient and promiscuous infection of cancer cell lines using a novel cell entry mechanism. 2020, 1 .
AMA StyleAlexander T. Baker, Gareth L Marlow, James A. Davies, Elise Moses, Rosie M. Mundy, David K. Cole, Pierre J. Rizkallah, Alan L. Parker. The fiber knob protein of human adenovirus type 49 mediates highly efficient and promiscuous infection of cancer cell lines using a novel cell entry mechanism. . 2020; ():1.
Chicago/Turabian StyleAlexander T. Baker; Gareth L Marlow; James A. Davies; Elise Moses; Rosie M. Mundy; David K. Cole; Pierre J. Rizkallah; Alan L. Parker. 2020. "The fiber knob protein of human adenovirus type 49 mediates highly efficient and promiscuous infection of cancer cell lines using a novel cell entry mechanism." , no. : 1.
Background The COVID-19 global pandemic has resulted in a plethora of guidance and opinion from surgical societies. A controversial area concerns the safety of surgically created smoke and the perceived potential higher risk in laparoscopic surgery. Methods The limited published evidence was analysed in combination with expert opinion. A review was undertaken of the novel coronavirus with regards to its hazards within surgical smoke and the procedures that could mitigate the potential risks to healthcare staff. Results Using existing knowledge of surgical smoke, a theoretical risk of virus transmission exists. Best practice should consider the operating room set-up, patient movement and operating theatre equipment when producing a COVID-19 operating protocol. The choice of energy device can affect the smoke produced, and surgeons should manage the pneumoperitoneum meticulously during laparoscopic surgery. Devices to remove surgical smoke, including extractors, filters and non-filter devices, are discussed in detail. Conclusion There is not enough evidence to quantify the risks of COVID-19 transmission in surgical smoke. However, steps can be undertaken to manage the potential hazards. The advantages of minimally invasive surgery may not need to be sacrificed in the current crisis.
N. G. Mowbray; J. Ansell; J. Horwood; J. Cornish; P. Rizkallah; A. Parker; P. Wall; A. Spinelli; J. Torkington. Safe management of surgical smoke in the age of COVID-19. British Journal of Surgery 2020, 107, 1406 -1413.
AMA StyleN. G. Mowbray, J. Ansell, J. Horwood, J. Cornish, P. Rizkallah, A. Parker, P. Wall, A. Spinelli, J. Torkington. Safe management of surgical smoke in the age of COVID-19. British Journal of Surgery. 2020; 107 (11):1406-1413.
Chicago/Turabian StyleN. G. Mowbray; J. Ansell; J. Horwood; J. Cornish; P. Rizkallah; A. Parker; P. Wall; A. Spinelli; J. Torkington. 2020. "Safe management of surgical smoke in the age of COVID-19." British Journal of Surgery 107, no. 11: 1406-1413.
Martin J. Scurr; Alexander Greenshields-Watson; Emma Campbell; Michelle S. Somerville; Yuan Chen; Sarah L. Hulin-Curtis; Stephanie E.A. Burnell; James Davies; Michael M. Davies; Rachel Hargest; Simon Phillips; Adam D. Christian; Kevin E. Ashelford; Robert Andrews; Alan L. Parker; Richard J. Stanton; Awen Gallimore; Andrew Godkin. Cancer Antigen Discovery Is Enabled by RNA Sequencing of Highly Purified Malignant and Nonmalignant Cells. Clinical Cancer Research 2020, 26, 3360 -3370.
AMA StyleMartin J. Scurr, Alexander Greenshields-Watson, Emma Campbell, Michelle S. Somerville, Yuan Chen, Sarah L. Hulin-Curtis, Stephanie E.A. Burnell, James Davies, Michael M. Davies, Rachel Hargest, Simon Phillips, Adam D. Christian, Kevin E. Ashelford, Robert Andrews, Alan L. Parker, Richard J. Stanton, Awen Gallimore, Andrew Godkin. Cancer Antigen Discovery Is Enabled by RNA Sequencing of Highly Purified Malignant and Nonmalignant Cells. Clinical Cancer Research. 2020; 26 (13):3360-3370.
Chicago/Turabian StyleMartin J. Scurr; Alexander Greenshields-Watson; Emma Campbell; Michelle S. Somerville; Yuan Chen; Sarah L. Hulin-Curtis; Stephanie E.A. Burnell; James Davies; Michael M. Davies; Rachel Hargest; Simon Phillips; Adam D. Christian; Kevin E. Ashelford; Robert Andrews; Alan L. Parker; Richard J. Stanton; Awen Gallimore; Andrew Godkin. 2020. "Cancer Antigen Discovery Is Enabled by RNA Sequencing of Highly Purified Malignant and Nonmalignant Cells." Clinical Cancer Research 26, no. 13: 3360-3370.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Michael D. Crowther; Garry Dolton; Mateusz Legut; Marine E. Caillaud; Angharad Lloyd; Meriem Attaf; Sarah A. E. Galloway; Cristina Rius; Colin P. Farrell; Barbara Szomolay; Ann Ager; Alan L. Parker; Anna Fuller; Marco Donia; James McCluskey; Jamie Rossjohn; Inge Marie Svane; John D. Phillips; Andrew K. Sewell. Author Correction: Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nature Immunology 2020, 21, 695 -695.
AMA StyleMichael D. Crowther, Garry Dolton, Mateusz Legut, Marine E. Caillaud, Angharad Lloyd, Meriem Attaf, Sarah A. E. Galloway, Cristina Rius, Colin P. Farrell, Barbara Szomolay, Ann Ager, Alan L. Parker, Anna Fuller, Marco Donia, James McCluskey, Jamie Rossjohn, Inge Marie Svane, John D. Phillips, Andrew K. Sewell. Author Correction: Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nature Immunology. 2020; 21 (6):695-695.
Chicago/Turabian StyleMichael D. Crowther; Garry Dolton; Mateusz Legut; Marine E. Caillaud; Angharad Lloyd; Meriem Attaf; Sarah A. E. Galloway; Cristina Rius; Colin P. Farrell; Barbara Szomolay; Ann Ager; Alan L. Parker; Anna Fuller; Marco Donia; James McCluskey; Jamie Rossjohn; Inge Marie Svane; John D. Phillips; Andrew K. Sewell. 2020. "Author Correction: Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1." Nature Immunology 21, no. 6: 695-695.
Human leukocyte antigen (HLA)-independent, T cell–mediated targeting of cancer cells would allow immune destruction of malignancies in all individuals. Here, we use genome-wide CRISPR–Cas9 screening to establish that a T cell receptor (TCR) recognized and killed most human cancer types via the monomorphic MHC class I-related protein, MR1, while remaining inert to noncancerous cells. Unlike mucosal-associated invariant T cells, recognition of target cells by the TCR was independent of bacterial loading. Furthermore, concentration-dependent addition of vitamin B-related metabolite ligands of MR1 reduced TCR recognition of cancer cells, suggesting that recognition occurred via sensing of the cancer metabolome. An MR1-restricted T cell clone mediated in vivo regression of leukemia and conferred enhanced survival of NSG mice. TCR transfer to T cells of patients enabled killing of autologous and nonautologous melanoma. These findings offer opportunities for HLA-independent, pan-cancer, pan-population immunotherapies. Identifying selective tumor-associated molecules that can act as targets for T cells is a major goal of immunotherapy. Sewell and colleagues demonstrate that the nonclassical MHC molecule MR1 is expressed on a wide variety of cancer types and can be targeted by conventional T cells.
Michael Crowther; Garry Dolton; Mateusz Legut; Marine Caillaud; Angharad Lloyd; Meriem Attaf; Sarah A. E. Galloway; Cristina Rius; Colin P. Farrell; Barbara Szomolay; Ann Ager; Alan L. Parker; Anna Fuller; Marco Donia; James McCluskey; Jamie Rossjohn; Inge Marie Svane; John D. Phillips; Andrew K. Sewell. Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nature Immunology 2020, 21, 178 -185.
AMA StyleMichael Crowther, Garry Dolton, Mateusz Legut, Marine Caillaud, Angharad Lloyd, Meriem Attaf, Sarah A. E. Galloway, Cristina Rius, Colin P. Farrell, Barbara Szomolay, Ann Ager, Alan L. Parker, Anna Fuller, Marco Donia, James McCluskey, Jamie Rossjohn, Inge Marie Svane, John D. Phillips, Andrew K. Sewell. Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1. Nature Immunology. 2020; 21 (2):178-185.
Chicago/Turabian StyleMichael Crowther; Garry Dolton; Mateusz Legut; Marine Caillaud; Angharad Lloyd; Meriem Attaf; Sarah A. E. Galloway; Cristina Rius; Colin P. Farrell; Barbara Szomolay; Ann Ager; Alan L. Parker; Anna Fuller; Marco Donia; James McCluskey; Jamie Rossjohn; Inge Marie Svane; John D. Phillips; Andrew K. Sewell. 2020. "Genome-wide CRISPR–Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1." Nature Immunology 21, no. 2: 178-185.
Oncolytic virotherapies (OV) based on human adenoviral (HAdV) vectors hold significant promise for the treatment of advanced ovarian cancers where local, intraperitoneal delivery to tumour metastases is feasible, bypassing many complexities associated with intravascular delivery. The efficacy of HAdV-C5-based OV is hampered by a lack of tumour selectivity, where the primary receptor, hCAR, is commonly downregulated during malignant transformation. Conversely, folate receptor alpha (FRα) is highly expressed on ovarian cancer cells, providing a compelling target for tumour selective delivery of virotherapies. Here, we identify high-affinity FRα-binding oligopeptides for genetic incorporation into HAdV-C5 vectors. Biopanning identified a 12-mer linear peptide, DWSSWVYRDPQT, and two 7-mer cysteine-constrained peptides, CIGNSNTLC and CTVRTSAEC that bound FRα in the context of the phage particle. Synthesised lead peptide, CTVRTSAEC, bound specifically to FRα and could be competitively inhibited with folic acid. To assess the capacity of the elucidated FRα-binding oligopeptides to target OV to FRα, we genetically incorporated the peptides into the HAdV-C5 fiber-knob HI loop including in vectors genetically ablated for hCAR interactions. Unfortunately, the recombinant vectors failed to efficiently target transduction via FRα due to defective intracellular trafficking following entry via FRα, indicating that whilst the peptides identified may have potential for applications for targeted drug delivery, they require additional refinement for targeted virotherapy applications.
Sarah Curtis; James Davies; Davor Nestić; Emily A. Bates; Alexander Baker; Tabitha G. Cunliffe; Dragomira Majhen; John D. Chester; Alan L. Parker. Identification of folate receptor α (FRα) binding oligopeptides and their evaluation for targeted virotherapy applications. Cancer Gene Therapy 2020, 27, 785 -798.
AMA StyleSarah Curtis, James Davies, Davor Nestić, Emily A. Bates, Alexander Baker, Tabitha G. Cunliffe, Dragomira Majhen, John D. Chester, Alan L. Parker. Identification of folate receptor α (FRα) binding oligopeptides and their evaluation for targeted virotherapy applications. Cancer Gene Therapy. 2020; 27 (10):785-798.
Chicago/Turabian StyleSarah Curtis; James Davies; Davor Nestić; Emily A. Bates; Alexander Baker; Tabitha G. Cunliffe; Dragomira Majhen; John D. Chester; Alan L. Parker. 2020. "Identification of folate receptor α (FRα) binding oligopeptides and their evaluation for targeted virotherapy applications." Cancer Gene Therapy 27, no. 10: 785-798.
Early phase clinical trials have demonstrated good therapeutic index for oncolytic adenoviruses in patients with solid tumours when administered intratumorally, resulting in local tumour elimination. Entrapment and binding of adenovirus to erythrocytes, blood factors, and neutralising antibodies have prevented efficient systemic delivery and targeting of distant lesions in the clinic. We previously generated the novel replication-selective Ad-3∆-A20T to improve tumour targeting by increasing the viral dose at distant sites. Here, we developed a protocol to directly radiolabel the virus for rapid and sensitive detection by single-photon emitted computed tomography (SPECT/CT) providing a convenient method for determining biodistribution following intravenous administration in murine models. Longitudinal whole-body scans, demonstrated efficient viral uptake in pancreatic Suit-2 and Panc04.03 xenografts with trace amounts of 125I-Ad-3∆-A20T up to 48 h after tail vein delivery. Hepatic and splenic radioactivity decreased over time. Analysis of tissues harvested at the end of the study, confirmed potency and selectivity of mutant viruses. Ad-3∆-A20T-treated animals showed higher viral genome copy numbers and E1A gene expression in tumors than in liver and spleen compared to Ad5wt. Our direct radiolabeling approach, allows for immediate screening of novel oncolytic adenoviruses and selection of optimal viral genome alterations to generate improved mutants.
Y. K. Stella Man; Julie Foster; Elisabete Carapuça; James Davies; Alan L. Parker; Jane Sosabowski; Gunnel Halldén. Systemic delivery and SPECT/CT in vivo imaging of 125I-labelled oncolytic adenoviral mutants in models of pancreatic cancer. Scientific Reports 2019, 9, 1 -12.
AMA StyleY. K. Stella Man, Julie Foster, Elisabete Carapuça, James Davies, Alan L. Parker, Jane Sosabowski, Gunnel Halldén. Systemic delivery and SPECT/CT in vivo imaging of 125I-labelled oncolytic adenoviral mutants in models of pancreatic cancer. Scientific Reports. 2019; 9 (1):1-12.
Chicago/Turabian StyleY. K. Stella Man; Julie Foster; Elisabete Carapuça; James Davies; Alan L. Parker; Jane Sosabowski; Gunnel Halldén. 2019. "Systemic delivery and SPECT/CT in vivo imaging of 125I-labelled oncolytic adenoviral mutants in models of pancreatic cancer." Scientific Reports 9, no. 1: 1-12.
Adenoviruses are clinically important agents. They cause respiratory distress, gastroenteritis, and epidemic keratoconjunctivitis. As non-enveloped, double-stranded DNA viruses, they are easily manipulated, making them popular vectors for therapeutic applications, including vaccines. Species D adenovirus type 26 (HAdV-D26) is both a cause of EKC and other diseases and a promising vaccine vector. HAdV-D26–derived vaccines are under investigation as protective platforms against HIV, Zika, and respiratory syncytial virus infections and are in phase 3 clinical trials for Ebola. We recently demonstrated that HAdV-D26 does not use CD46 or Desmoglein-2 as entry receptors, while the putative interaction with coxsackie and adenovirus receptor is low affinity and unlikely to represent the primary cell receptor. Here, we establish sialic acid as a primary entry receptor used by HAdV-D26. We demonstrate that removal of cell surface sialic acid inhibits HAdV-D26 infection, and provide a high-resolution crystal structure of HAdV-D26 fiber-knob in complex with sialic acid.
Alexander T. Baker; Rosie M. Mundy; James A. Davies; Pierre J. Rizkallah; Alan L. Parker. Human adenovirus type 26 uses sialic acid–bearing glycans as a primary cell entry receptor. Science Advances 2019, 5, eaax3567 .
AMA StyleAlexander T. Baker, Rosie M. Mundy, James A. Davies, Pierre J. Rizkallah, Alan L. Parker. Human adenovirus type 26 uses sialic acid–bearing glycans as a primary cell entry receptor. Science Advances. 2019; 5 (9):eaax3567.
Chicago/Turabian StyleAlexander T. Baker; Rosie M. Mundy; James A. Davies; Pierre J. Rizkallah; Alan L. Parker. 2019. "Human adenovirus type 26 uses sialic acid–bearing glycans as a primary cell entry receptor." Science Advances 5, no. 9: eaax3567.
Adenoviruses are clinically important agents. They cause respiratory distress, gastroenteritis, and epidemic keratoconjunctivitis (EKC). As non-enveloped, double stranded DNA viruses, they are easily manipulated, making them popular vectors for therapeutic applications, including vaccines. Species D adenovirus serotype 26 (HAdV-D26) is both a cause of EKC and other disease, and a promising vaccine vector. HAdV-D26 derived vaccines are under investigation as protective platforms against HIV, Zika, RSV infections and are in Phase-III clinical trials for Ebola.We recently demonstrated that HAdV-D26 does not utilise CD46 or desmoglein 2 as entry receptors, whilst the putative interaction with Coxsackie and Adenovirus Receptor (CAR) is low affinity and unlikely to represent the primary cell receptor.Here, we definitively establish sialic acid as the primary entry receptor utilised by HAdV-D26. We demonstrate removal of cell surface sialic acid inhibits HAdV-D26 infection and provide a high-resolution crystal structure of HAdV-D26 fiber-knob in complex with sialic acid.
Alexander T. Baker; Rosie Mundy; James Davies; Pierre J. Rizkallah; Alan L Parker. Adenovirus serotype 26 utilises sialic acid bearing glycans as a primary cell entry receptor. 2019, 580076 .
AMA StyleAlexander T. Baker, Rosie Mundy, James Davies, Pierre J. Rizkallah, Alan L Parker. Adenovirus serotype 26 utilises sialic acid bearing glycans as a primary cell entry receptor. . 2019; ():580076.
Chicago/Turabian StyleAlexander T. Baker; Rosie Mundy; James Davies; Pierre J. Rizkallah; Alan L Parker. 2019. "Adenovirus serotype 26 utilises sialic acid bearing glycans as a primary cell entry receptor." , no. : 580076.
Adenovirus based vectors are of increasing importance for wide ranging therapeutic applications. As vaccines, vectors derived from human adenovirus species D serotypes 26 and 48 (HAdV-D26/48) are demonstrating promising efficacy as protective platforms against infectious diseases. Significant clinical progress has been made, yet definitive studies underpinning mechanisms of entry, infection, and receptor usage are currently lacking. Here, we perform structural and biological analysis of the receptor binding fiber-knob protein of HAdV-D26/48, reporting crystal structures, and modelling putative interactions with two previously suggested attachment receptors, CD46 and Coxsackie and Adenovirus Receptor (CAR). We provide evidence of a low affinity interaction with CAR, with modelling suggesting affinity is attenuated through extended, semi-flexible loop structures, providing steric hindrance. Conversely, in silico and in vitro experiments are unable to provide evidence of interaction between HAdV-D26/48 fiber-knob with CD46, or with Desmoglein 2. Our findings provide insight into the cell-virus interactions of HAdV-D26/48, with important implications for the design and engineering of optimised Ad-based therapeutics. Adenovirus based (AdV) vectors are promising platforms for therapeutics and vaccines, but receptor usage of serotypes in clinical development remains unclear. Here, based on crystal structures and modeling, Baker et al. show that HAdV-D26/48 fiber knob protein interacts weakly with CAR but not with CD46 or DSG2.
Alexander T. Baker; Alexander Greenshields-Watson; Lynda Coughlan; James A. Davies; Hanni Uusi-Kerttula; David K. Cole; Pierre J. Rizkallah; Alan L. Parker. Diversity within the adenovirus fiber knob hypervariable loops influences primary receptor interactions. Nature Communications 2019, 10, 1 -14.
AMA StyleAlexander T. Baker, Alexander Greenshields-Watson, Lynda Coughlan, James A. Davies, Hanni Uusi-Kerttula, David K. Cole, Pierre J. Rizkallah, Alan L. Parker. Diversity within the adenovirus fiber knob hypervariable loops influences primary receptor interactions. Nature Communications. 2019; 10 (1):1-14.
Chicago/Turabian StyleAlexander T. Baker; Alexander Greenshields-Watson; Lynda Coughlan; James A. Davies; Hanni Uusi-Kerttula; David K. Cole; Pierre J. Rizkallah; Alan L. Parker. 2019. "Diversity within the adenovirus fiber knob hypervariable loops influences primary receptor interactions." Nature Communications 10, no. 1: 1-14.