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Viral vector engineer researching clinical translational oncolytic and gene transfer viruses.
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.
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.
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.
Recombinant vesicular stomatitis virus (VSV)-fusion and hemagglutinin (FH) was developed by substituting the promiscuous VSV-G glycoprotein (G) gene in the backbone of VSV with genes encoding for the measles virus envelope proteins F and H. Hybrid VSV-FH exhibited a multifaceted mechanism of cancer-cell killing and improved neurotolerability over parental VSV in preclinical studies. In this study, we evaluated VSV-FH in vitro and in vivo in models of hepatobiliary and pancreatic cancers. Our results indicate that high intrahepatic doses of VSV-FH did not result in any significant toxicity and were well tolerated by transgenic mice expressing the measles virus receptor CD46. Furthermore, a single intratumoral treatment with VSV-FH yielded improved survival and complete tumor regressions in a proportion of mice in the Hep3B hepatocellular carcinoma model but not in mice xenografted with BxPC-3 pancreatic cancer cells. Our preliminary findings indicate that VSV-FH can induce potent oncolysis in hepatocellular and pancreatic cancer cell lines with concordant results in vivo in hepatocellular cancer and discordant in pancreatic cancer without the VSV-mediated toxic effects previously observed in laboratory animals. Further study of VSV-FH as an oncolytic virotherapy is warranted in hepatocellular carcinoma and pancreatic cancer to understand broader applicability and mechanisms of sensitivity and resistance.
Bolni Marius Nagalo; Camilo Ayala Breton; Yumei Zhou; Mansi Arora; James M. Bogenberger; Oumar Barro; Michael B. Steele; Nathan J. Jenks; Alexander Baker; Dan G. Duda; Lewis Rowland Roberts; Stephen J. Russell; Kah Whye Peng; Mitesh J. Borad. Oncolytic Virus with Attributes of Vesicular Stomatitis Virus and Measles Virus in Hepatobiliary and Pancreatic Cancers. Molecular Therapy - Oncolytics 2020, 18, 546 -555.
AMA StyleBolni Marius Nagalo, Camilo Ayala Breton, Yumei Zhou, Mansi Arora, James M. Bogenberger, Oumar Barro, Michael B. Steele, Nathan J. Jenks, Alexander Baker, Dan G. Duda, Lewis Rowland Roberts, Stephen J. Russell, Kah Whye Peng, Mitesh J. Borad. Oncolytic Virus with Attributes of Vesicular Stomatitis Virus and Measles Virus in Hepatobiliary and Pancreatic Cancers. Molecular Therapy - Oncolytics. 2020; 18 ():546-555.
Chicago/Turabian StyleBolni Marius Nagalo; Camilo Ayala Breton; Yumei Zhou; Mansi Arora; James M. Bogenberger; Oumar Barro; Michael B. Steele; Nathan J. Jenks; Alexander Baker; Dan G. Duda; Lewis Rowland Roberts; Stephen J. Russell; Kah Whye Peng; Mitesh J. Borad. 2020. "Oncolytic Virus with Attributes of Vesicular Stomatitis Virus and Measles Virus in Hepatobiliary and Pancreatic Cancers." Molecular Therapy - Oncolytics 18, no. : 546-555.
Viruses as cancer therapies have attracted attention since the 19th century. Scientists observation that viruses can preferentially lyse cancer cells rather than healthy cells, created the field of oncolytic virology. Like other therapeutic strategies, oncolytic virotherapy has challenges, such as penetration into tumor bulk, anti-viral immune responses, off-target infection, adverse conditions in the tumor microenvironment, and the lack of specific predictive and therapeutic biomarkers. Whilst much progress has been made, as highlighted by the first Food and Drug Administration approval of an oncolytic virus (T-VEC) in 2015, addressing these issues remains a significant hurdle. Here we discuss different types of oncolytic viruses, their application in clinical trials, and finally challenges faced by the field of oncolytic virotherapy and strategies to overcome them.
Nasser Hashemi Goradel; Alexander Baker; Arash Arashkia; Nasim Ebrahimi; Sajjad Ghorghanlu; Babak Negahdari. Oncolytic virotherapy: Challenges and solutions. Current Problems in Cancer 2020, 45, 100639 .
AMA StyleNasser Hashemi Goradel, Alexander Baker, Arash Arashkia, Nasim Ebrahimi, Sajjad Ghorghanlu, Babak Negahdari. Oncolytic virotherapy: Challenges and solutions. Current Problems in Cancer. 2020; 45 (1):100639.
Chicago/Turabian StyleNasser Hashemi Goradel; Alexander Baker; Arash Arashkia; Nasim Ebrahimi; Sajjad Ghorghanlu; Babak Negahdari. 2020. "Oncolytic virotherapy: Challenges and solutions." Current Problems in Cancer 45, no. 1: 100639.
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.
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.
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.
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 performed 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 silicoandin vitroexperiments are unable to provide evidence of interaction between HAdV-D26/48 fiber-knob with CD46, or with Desmoglein 2. Our findings provide new insight to the cell-virus interactions of HAdV-D26/48, with important implications for the design and engineering of optimised Ad-based therapeutics.
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. 2018, 406819 .
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. . 2018; ():406819.
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. 2018. "Diversity within the adenovirus fiber knob hypervariable loops influences primary receptor interactions." , no. : 406819.
The licensing of talimogene laherparepvec (T-Vec) represented a landmark moment for oncolytic virotherapy, since it provided unequivocal evidence for the long-touted potential of genetically modified replicating viruses as anti-cancer agents. Whilst T-Vec is promising as a locally delivered virotherapy, especially in combination with immune-checkpoint inhibitors, the quest continues for a virus capable of specific tumour cell killing via systemic administration. One candidate is oncolytic adenovirus (Ad); it’s double stranded DNA genome is easily manipulated and a wide range of strategies and technologies have been employed to empower the vector with improved pharmacokinetics and tumour targeting ability. As well characterised clinical and experimental agents, we have detailed knowledge of adenoviruses’ mechanisms of pathogenicity, supported by detailed virological studies and in vivo interactions. In this review we highlight the strides made in the engineering of bespoke adenoviral vectors to specifically infect, replicate within, and destroy tumour cells. We discuss how mutations in genes regulating adenoviral replication after cell entry can be used to restrict replication to the tumour, and summarise how detailed knowledge of viral capsid interactions enable rational modification to eliminate native tropisms, and simultaneously promote active uptake by cancerous tissues. We argue that these designer-viruses, exploiting the viruses natural mechanisms and regulated at every level of replication, represent the ideal platforms for local overexpression of therapeutic transgenes such as immunomodulatory agents. Where T-Vec has paved the way, Ad-based vectors now follow. The era of designer oncolytic virotherapies looks decidedly as though it will soon become a reality.
Alexander T. Baker; Carmen Aguirre-Hernández; Gunnel Hallden; Alan L. Parker. Designer Oncolytic Adenovirus: Coming of Age. 2018, 1 .
AMA StyleAlexander T. Baker, Carmen Aguirre-Hernández, Gunnel Hallden, Alan L. Parker. Designer Oncolytic Adenovirus: Coming of Age. . 2018; ():1.
Chicago/Turabian StyleAlexander T. Baker; Carmen Aguirre-Hernández; Gunnel Hallden; Alan L. Parker. 2018. "Designer Oncolytic Adenovirus: Coming of Age." , no. : 1.
The licensing of talimogene laherparepvec (T-Vec) represented a landmark moment for oncolytic virotherapy, since it provided unequivocal evidence for the long-touted potential of genetically modified replicating viruses as anti-cancer agents. Whilst T-Vec is promising as a locally delivered virotherapy, especially in combination with immune-checkpoint inhibitors, the quest continues for a virus capable of specific tumour cell killing via systemic administration. One candidate is oncolytic adenovirus (Ad); it’s double stranded DNA genome is easily manipulated and a wide range of strategies and technologies have been employed to empower the vector with improved pharmacokinetics and tumour targeting ability. As well characterised clinical and experimental agents, we have detailed knowledge of adenoviruses’ mechanisms of pathogenicity, supported by detailed virological studies and in vivo interactions. In this review we highlight the strides made in the engineering of bespoke adenoviral vectors to specifically infect, replicate within, and destroy tumour cells. We discuss how mutations in genes regulating adenoviral replication after cell entry can be used to restrict replication to the tumour, and summarise how detailed knowledge of viral capsid interactions enable rational modification to eliminate native tropisms, and simultaneously promote active uptake by cancerous tissues. We argue that these designer-viruses, exploiting the viruses natural mechanisms and regulated at every level of replication, represent the ideal platforms for local overexpression of therapeutic transgenes such as immunomodulatory agents. Where T-Vec has paved the way, Ad-based vectors now follow. The era of designer oncolytic virotherapies looks decidedly as though it will soon become a reality.
Alexander Baker; Carmen Aguirre-Hernández; Gunnel Halldén; Alan Parker. Designer Oncolytic Adenovirus: Coming of Age. Cancers 2018, 10, 201 .
AMA StyleAlexander Baker, Carmen Aguirre-Hernández, Gunnel Halldén, Alan Parker. Designer Oncolytic Adenovirus: Coming of Age. Cancers. 2018; 10 (6):201.
Chicago/Turabian StyleAlexander Baker; Carmen Aguirre-Hernández; Gunnel Halldén; Alan Parker. 2018. "Designer Oncolytic Adenovirus: Coming of Age." Cancers 10, no. 6: 201.
Purpose: Virotherapies are maturing in the clinical setting. Adenoviruses (Ad) are excellent vectors for the manipulability and tolerance of transgenes. Poor tumor selectivity, off-target sequestration, and immune inactivation hamper clinical efficacy. We sought to completely redesign Ad5 into a refined, tumor-selective virotherapy targeted to αvβ6 integrin, which is expressed in a range of aggressively transformed epithelial cancers but nondetectable in healthy tissues. Experimental Design: Ad5NULL-A20 harbors mutations in each major capsid protein to preclude uptake via all native pathways. Tumor-tropism via αvβ6 targeting was achieved by genetic insertion of A20 peptide (NAVPNLRGDLQVLAQKVART) within the fiber knob protein. The vector's selectivity in vitro and in vivo was assessed. Results: The tropism-ablating triple mutation completely blocked all native cell entry pathways of Ad5NULL-A20 via coxsackie and adenovirus receptor (CAR), αvβ3/5 integrins, and coagulation factor 10 (FX). Ad5NULL-A20 efficiently and selectively transduced αvβ6+ cell lines and primary clinical ascites-derived EOC ex vivo, including in the presence of preexisting anti-Ad5 immunity. In vivo biodistribution of Ad5NULL-A20 following systemic delivery in non–tumor-bearing mice was significantly reduced in all off-target organs, including a remarkable 107-fold reduced genome accumulation in the liver compared with Ad5. Tumor uptake, transgene expression, and efficacy were confirmed in a peritoneal SKOV3 xenograft model of human EOC, where oncolytic Ad5NULL-A20–treated animals demonstrated significantly improved survival compared with those treated with oncolytic Ad5. Conclusions: Oncolytic Ad5NULL-A20 virotherapies represent an excellent vector for local and systemic targeting of αvβ6-overexpressing cancers and exciting platforms for tumor-selective overexpression of therapeutic anticancer modalities, including immune checkpoint inhibitors. Clin Cancer Res; 24(17); 4215–24. ©2018 AACR.
Hanni Uusi-Kerttula; James Davies; Jill M. Thompson; Phonphimon Wongthida; Laura Evgin; Kevin G. Shim; Angela Bradshaw; Alexander T. Baker; Pierre Rizkallah; Rachel Jones; Louise Hanna; Emma Hudson; Richard G. Vile; John D. Chester; Alan L. Parker. Ad5NULL-A20: A Tropism-Modified, αvβ6 Integrin-Selective Oncolytic Adenovirus for Epithelial Ovarian Cancer Therapies. Clinical Cancer Research 2018, 24, 4215 -4224.
AMA StyleHanni Uusi-Kerttula, James Davies, Jill M. Thompson, Phonphimon Wongthida, Laura Evgin, Kevin G. Shim, Angela Bradshaw, Alexander T. Baker, Pierre Rizkallah, Rachel Jones, Louise Hanna, Emma Hudson, Richard G. Vile, John D. Chester, Alan L. Parker. Ad5NULL-A20: A Tropism-Modified, αvβ6 Integrin-Selective Oncolytic Adenovirus for Epithelial Ovarian Cancer Therapies. Clinical Cancer Research. 2018; 24 (17):4215-4224.
Chicago/Turabian StyleHanni Uusi-Kerttula; James Davies; Jill M. Thompson; Phonphimon Wongthida; Laura Evgin; Kevin G. Shim; Angela Bradshaw; Alexander T. Baker; Pierre Rizkallah; Rachel Jones; Louise Hanna; Emma Hudson; Richard G. Vile; John D. Chester; Alan L. Parker. 2018. "Ad5NULL-A20: A Tropism-Modified, αvβ6 Integrin-Selective Oncolytic Adenovirus for Epithelial Ovarian Cancer Therapies." Clinical Cancer Research 24, no. 17: 4215-4224.
Holomycin and its derivatives belong to a class of broad-spectrum antibacterial natural products containing a rare dithiolopyrrolone heterobicyclic scaffold. The antibacterial mechanism of dithiolopyrrolone compounds has been attributed to the inhibition of bacterial RNA polymerase activities, although the exact mode of action has not been established in vitro. Some dithiopyrrolone derivatives display potent anticancer activities. Recently the biosynthetic gene cluster of holomycin has been identified and characterized in Streptomyces clavuligerus. Here we report that the fish pathogen Yersinia ruckeri is a holomycin producer, as evidenced through genome mining, chemical isolation, and structural elucidation as well as genetic manipulation. We also identified a unique regulatory gene hom15 at one end of the gene cluster encoding a cold-shock-like protein that likely regulates the production of holomycin in low cultivation temperatures. Inactivation of hom15 resulted in a significant loss of holomycin production. Finally, gene disruption of an RNA methyltransferase gene hom12 resulted in the sensitivity of the mutant toward holomycin. A complementation experiment of hom12 restored the resistance against holomycin. Although the wild-type Escherichia coli BL21(DE3) Gold is susceptible to holomycin, the mutant harboring hom12 showed tolerance toward holomycin. High resolution liquid chromatography (LC)-ESI/MS analysis of digested RNA fragments demonstrated that the wild-type Y. ruckeri and E. coli harboring hom12 contain a methylated RNA fragment, whereas the mutated Y. ruckeri and the wild-type E. coli only contain normal non-methylated RNA fragments. Taken together, our results strongly suggest that this putative RNA methyltransferase Hom12 is the self-resistance protein that methylates the RNA of Y. ruckeri to reduce the cytotoxic effect of holomycin during holomycin production.
Zhiwei Qin; Alexander Baker; Andrea Raab; Sheng Huang; Tiehui Wang; Yi Yu; Marcel Jaspars; Christopher J. Secombes; Hai Deng. The Fish Pathogen Yersinia ruckeri Produces Holomycin and Uses an RNA Methyltransferase for Self-resistance. Journal of Biological Chemistry 2013, 288, 14688 -14697.
AMA StyleZhiwei Qin, Alexander Baker, Andrea Raab, Sheng Huang, Tiehui Wang, Yi Yu, Marcel Jaspars, Christopher J. Secombes, Hai Deng. The Fish Pathogen Yersinia ruckeri Produces Holomycin and Uses an RNA Methyltransferase for Self-resistance. Journal of Biological Chemistry. 2013; 288 (21):14688-14697.
Chicago/Turabian StyleZhiwei Qin; Alexander Baker; Andrea Raab; Sheng Huang; Tiehui Wang; Yi Yu; Marcel Jaspars; Christopher J. Secombes; Hai Deng. 2013. "The Fish Pathogen Yersinia ruckeri Produces Holomycin and Uses an RNA Methyltransferase for Self-resistance." Journal of Biological Chemistry 288, no. 21: 14688-14697.