Models of virusinduced carcinogenesis and oncolytic viruses. Oncolytic herpes simplex virus engineering and preparation construction of capsidmodified adenoviruses by recombination in yeast and purification by iodixanolgradient construction of targeted and armed oncolytic adenoviruses syrian hamster tumor model to study oncolytic ad5based vectors adenoviral gene expression and replication in. The present invention relates to recombinant oncolytic viruses. Ccl5armed oncolytic virus augments ccr5engineered nk cell. Human mesenchymal stromal cells deliver systemic oncolytic. The idea for using these viruses as a cancer therapy is not a new one, and is based on patients demonstrating tumor regression after a natural viral infection. Conclusions overall, oncolytic adenovirus ad53e2fd24htnfaireshil2 was able to rewire the ovarian tumor microenvironment to accommodate heightened antitumor til reactivity.
Besides the direct killing potential oncolysis, what makes ov administration attractive for the present cancer immunotherapeutic scenario is the capacity to induce two new. Since the 1960s oncolytic viruses ovs have been a target of research as a therapeutic modality for cancer. Concepts for the production of viruses and viral vectors. Comparison of clinically relevant oncolytic virus platforms. Ovs and their resultant immunological events can act at both primary and metastatic sites. Methods and protocols provides a wide collection of protocols employed in various levels of herpes virus research, including basic protocols on growing viruses in cell culture and cloning, manipulating and preparing viral dna. Ovs can revert cancerassociated immune suppression and initiate clinically meaningful antitumor immune responses. The pancancerspecific oncolytic potency of nov was. Complex multistep processes are required in both cases, but the low virus titers in batch cultures and the temperature sensitivity of the virus particles limit the production scale.
Oncolytic viruses are promising for cancer therapeutics. This treatment involves a herpes virus that has been engineered to be less likely to infect healthy cells as well as cause infected cancer cells to produce the immunestimulating gmcsf protein. Oncolytic viruses are thought not only to cause direct destruction of the tumour cells, but also to stimulate host. However, in contrast to human studies, only one clinical. Oncolytic viruses are biologic agents able to selectively infect and destroy cancer cells while sparing the normal ones. More recently, some viruses, the socalled oncolytic viruses, have been used as agents against cancer.
However, their antitumoral potential has remained largely unexplored, due to safety concerns and some limitations in the techniques to manipulate viruses. This is achieved by producing cytotoxicityenhancing proteins andor eliciting an antitumor immune response. Current standards of care are also associated with significant morbidities, giving impetus for the development of new treatments. The ability to ascertain the infectability of live tissue specimens obtained from patients prior to treatment is a unique advantage of this therapeutic approach. The use of viruses as oncolytic agents ovs was considered in the past, while only recently ovs revealed a connection with immunotherapy. Dear colleagues, this special issue, oncolytic viruses as a novel form of immunotherapy for cancer, will mainly focus on oncolytic viruses ovs and their induced antitumor immunity for cancer immunotherapy. Oncolytic viruses range in size and complexity from large, doublestranded dna viruses such as vaccinia 190 kilobase kb 17 and hsv1 152 kb 18 to the tiny parvovirus h1 5kb linear, singlestranded dna. This volume that has been edited by pioneers in the field of oncolytic virus therapy represents an attempt to focus on specific methodologies and protocols that are currently being widely applied in this area of research. The preexisting neutralizing antibody in patients is a big challenge for oncolytic efficacy of ovs. Inhibition of pyruvate dehydrogenase kinase enhances the. Directed evolution generates a novel oncolytic virus for. A panel of diverse viruses, often genetically engineered, has advanced to clinical investigation, including phase 3 studies. Oncolytic viruses ovs constitute a new and promising immunotherapeutic approach toward cancer treatment. Immune checkpoint inhibitors have shown great promise for cancer therapy, but they do not treat all cancers, and neither breast nor brain tumors are usually treatable with these drugs.
Oncolytic viruses have the potential to induce immunogenic cell death icd that may provoke potent and longlasting anticancer immunity. Oncolytic viruses which infect and kill tumour cells can also be genetically modified to express therapeutic genes that augment their anticancer activities. The ultimate aim is to design a virus which can effectively replicate within the host, specifically target and lyse tumor cells and induce robust, long lasting tumorspecific immunity. Oncolytic viruses genetically engineering the future of.
Despite recent advances in both surgery and chemoradiotherapy, mortality rates for advanced cancer remain high. Replicationcompetent oncolytic viruses ovs have been proven to be a potent anticancer weapon for clinical therapy. The notes sections at the end of each chapter are a particularly useful source of guidance that will assist researchers in establishing new methods or in troubleshooting problematic techniques. Development of a versatile oncolytic virus platform for. Directed evolution generates a novel oncolytic virus for the. Oncolytic viruses ovs are unique anticancer agents based on their pleotropic modes of action, which include, besides viral tumor cell lysis, activation of antitumor immunity. Mg1 preferentially infects and eliminates cell lines latently infected with human immunodeficiency virus type 1 hiv1. An oncolytic virus named imlygic tm manufactured by amgen recently received the stamp of approval from the fda making exciting headlines in the research community across the globe. This volume that has been edited by pioneers in the field of oncolytic virus therapy represents an attempt to focus on specific methodologies and protocols.
Oncolytic viruses ovs preferentially infect and kill cancer cells without harming normal cells. However, advances in the treatment of highgrade gliomas, and specifically glioblastoma multiforme gbm, are the result of novel oncolytic viral therapies. The major obstacle facing the current development of antitumor drugs is not their potency, but rather their low selectivity for tumor cells. Modifying oncolytic viruses to produce effective cancer therapies is challenging as encoding transgenes often attenuates virus activity or prevents systemic delivery in patients due to the risk of offtarget expression of transgenes in. Perspectives on immunotherapy via oncolytic viruses.
Different production methods for viral particle vp have been described in the literature,19, 20, 21, 22, 23 including the growth of producer cells in suspension. Sep 09, 2016 oncolytic virus therapy has recently been recognized as a promising new therapeutic approach for cancer treatment. Mar 31, 2016 viruses associated with cancer and model systems to study these mechanisms have established basic paradigms for viruscell interactions and contributed greatly to the understanding of diseases associated with biomedically important viruses. This particular oncolytic virus is based on herpes simplex virus type 1 and has been modified to include a gene that codes for gmcsf, a protein that stimulates the production of immune cells in the body. Besides the direct oncolytic activity, ovs are also effective in stimulating the body immune responses which can further help recogonize, control or. Intravenous administration affords the opportunity to treat both the primary tumour and any metastatic deposits simultaneously. Oncolytic viruses ovs are an emerging treatment option for many cancer types and have recently been the focus of extensive research aiming to develop their therapeutic potential.
Methods and protocols aims to ensure successful results in the further study of this vital field. Oncolytic virus therapy shows benefit in patients with. We then upregulated ccr5 and ccl5 with lentiviruses and oncolytic viruses in nk and tumor cells, respectively. Written in the highly successful methods in molecular biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, stepbystep, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. An oncolytic virus is defined as a genetically engineered or naturally occurring virus that can selectively replicate in and kill cancer cells without harming the normal tissues. More specifically, the present invention relates to recombinant oncolytic viruses expressing a heterologous b cell attractant polypeptide or a t cell attractant polypeptide.
Thus, the mechanisms that dictate the susceptibility of cancer cells to ovinduced cytotoxicity hold the key to their success in clinics. We and others have developed a number of strategies to further stimulate antitumor immunity and to. This therapy takes advantage of the natural propensity of most tumor cells to be infected by specific ovs. Such effects may improve the clinical effectiveness of act with tils in patients with advanced ovca. Backgroundviralmediated oncolysis is a novel cancer therapeutic approach with the potential to be more effective and less toxic than current therapies due to the agents selective growth and amplification in tumor cells.
Ovs have been explored as therapeutic cancer vaccines for quite a few decades. An oncolytic virus is a virus that preferentially infects and kills cancer cells. Methods and protocols describes the construction and. Here, we investigated whether cancer cell metabolism defines its susceptibility to ov and if ovinduced metabolic perturbations can be therapeutically targeted. The conventional method for creation of tumortargeted oncolytic viruses is based on homologous recombination. Tumorselectively replicating oncolytic viruses are promising tools for therapy of solid cancers and have been initially developed to achieve potent tumor lysis with acceptable side effects on healthy tissue. Embodiments of the present disclosure concern oncolytic viruses, such as vaccinia virus, for example, for the treatment of cancer, wherein the viruses encode an engager molecule having an activation domain that recognizes a cell molecule, such as cd3, for example, on t cells and an antigen recognition domain that recognizes a tumor antigen, such as epha2, her2, gd2, or glypican3, for example.
Intravenous delivery of oncolytic reovirus to brain tumor. May 15, 2017 the concept of oncolytic virus ovmediated cancer therapy has been shifted from an operational virotherapy paradigm to an immunotherapy. Medulloblastoma is the most common type of pediatric brain tumor. Molecular imaging of oncolytic viral therapy sciencedirect. Methods and protocols describes the construction and purification of capsidmodified adenoviruses as well as oncolytic adenoviruses, presents protocols for many individual virus species including engineering and preparation of oncolytic hsv, propagation, purification, and in vivo testing of oncolytic vsv, details properties. Food and drug administration fda approved the first oncolytic virus immunotherapy for the treatment of cancertvec for melanoma. Oncolytic viruses are thought not only to cause direct destruction of the tumour cells, but also to stimulate host antitumour immune system responses. Methods and protocols methods in molecular biology. Feb 19, 20 instead, this book should be regarded as a how i do it guide to practical science with oncolytic viruses.
However, in recent years, oncolytic viruses have been recognized as therapeutic vehicles exhibiting multipronged antitumoral activity. Frontiers oncolytic virusesinteraction of virus and. Jan 18, 2016 the use of oncolytic viruses is a new cancer treatment that is excellent at achieving just that. Oncolytic viral therapies are increasingly being explored for the treatment of diverse cancer types, most notably melanoma. Thus, we analyzed ccr5 expression on nk cells from patients with cancer and healthy donors. Direct oncolysis releases tumor antigens, and viral replication within the tumor microenvironment is a potent danger signal. Methods the ccr5ccl5 axis is critical in nk cell homing to tumor sites. Oncolytic viruses have made their mark on the cancer world as a potential. The deeptissue imaging methods have been explored with oncolytic viral. We are also exploring the mechanisms of local immunity at sites to intratumoral virus. Reverse genetics of rna viruses methods and protocols. In this approach it is important that a therapeutic transgene insertion site within the viral genome be identified that does not compromise the life cycle and therefore the potency of the virus.
Characterization of virusmediated immunogenic cancer cell. Previously, we hierarchically developed and fit a mathematical model to the experimental data in huang et al. Treatment of medulloblastoma with oncolytic measles viruses. Oncolytic viruses offer a rational alternative to solve the limitations of those immunotherapies, but the lack of headtohead comparisons between different virus platforms has slowed optimization of the approach. Since the first report of an engineered oncolytic virus, there has been a. Evaluation of oncolytic virus induced therapeutic tumor vaccination effects in murine tumor models. In an effort to block the neurotropism yet retain the antitumor potency of a primary oncolytic virus, a second virus, ndv, was employed as an agent of interference. Here, we developed a novel oncolytic vaccinia virus nov with the dual advantages of cancer selectivity and normal vessel reconstructive activity by replacing the viral thymidine kinase vtk and vaccinia growth factor vgf genes with genes encoding tnfrelated apoptosisinducing ligand trail and angiopoietin 1 ang1, respectively. The testing strategy for molecular variants may need to reflect both the nature and amount of the variants. Although numerous factors influence patient survival rates, more than 30% of all cases will ultimately be refractory to conventional therapies. While certain viruses have been known to have the ability to kill cancer cells, it is only from 1990 to 2010 that technological advances in viral culture methodology and molecular engineering have made clinical translation possible. Part of the methods in molecular biology book series mimb, volume 2058 log in to check access. Thus, ovs can be exploited for cancer gene therapies and immunotherapies alone or in. Infectivity assays and immunohistochemical analysis of tumor sections were carried out to detect virus and viral proteins in the tumor.
Keywords virotherapy vector design immunomonitoring mathematical modeling talimogene laherparepvec clinical translation. Oncolytic viruses ov such as reovirus preferentially infect and kill cancer cells. Oncolytic viruses ovs preferentially infect and lyse transformed cells, leaving normal cells relatively unharmed. Join researchgate to discover and stay uptodate with the latest research from leading experts in oncolytic viruses and many. In vivo estimation of oncolytic virus populations within. Seneca valley virus, a systemically deliverable oncolytic. The introduction of oncolytic viruses for cancer therapy has provided a novel avenue of therapy that is showing increasingly promising results.
Activation of the immune response against cancer cells is emerging as one of the main mechanisms of action of oncolytic viruses ov. Dec 16, 2015 the limited efficacy of current treatments against pancreatic cancer has prompted the search of new alternatives such as virotherapy. With few exceptions, current chemotherapy and radiotherapy protocols only obtain a slightly prolonged survival with severe adverse effects in. Oncolytic virusbased immunotherapies for hepatocellular. Graphene oxide arms oncolytic measles virus for improved. Oncolytic virus therapy cancer research institute cri. Jan 19, 2006 although gene therapy has huge potential for modern medicine, our enthusiasm for its powerful potential must not cloud our judgment about the dangers of using increasingly diverse, yet relatively untested, replicating viruses gene therapy is currently being studied in both the laboratory and the clinic in relation to many conditions, including cancer, heart disease, and autoimmune diseases. Oncolytic viruses are a group of viruses that selectively infect and kill cancer cells while sparing normal tissues. Methods and protocols methods in molecular biology, vol. Furthermore, they also stimulate host immune system to combat the tumor growth and promote tumor removal. The lab has established numerous models to explore how innate and adaptive immunity responds to oncolytic immunotherapy. There is a pressing need for novel therapeutic strategies. Tvec is an oncolytic virus, a virus that selectively replicates inside and kills cancer cells.
Pdf the propagation and quantification of two emerging. One promising oncolytic virus therapy is called tvec, and is currently in latestage clinical testing. Oncolytic viruses as engineering platforms for combination. Tumorselective, replication competent viruses that have antineoplastic effects. Apr 25, 2018 in this opinion article, twumasiboateng et al. This volume that has been edited by pioneers in the field of oncolytic virus therapy represents an attempt to focus on specific methodologies and protocols that are currently being widely applied. Perspectives on immunotherapy via oncolytic viruses infectious. Developing a minimally structured mathematical model of.
Therefore, a new generation of more potent oncolytic viruses needs to be exploited, following our better understanding of the complex interactions between the tumor, its microenvironment, the virus, and the host immune response. To date, these agents have been highly safe in patients but have generally fallen short of their expected therapeutic value as monotherapies. Since the first report of an engineered oncolytic virus, there has been a continuing and steady increase of interest in the field, and while bench research remains vital for the translation of. Oncolytic viruses have been engineered to selectively infect and spread within the tumor cell population, leading to cancer cell death due to a variety of mechanisms including direct cell lysis, celltocell fusion, expression of. They lack crossresistance with existing therapies, and the acceptable safety profile of ovs has been demonstrated in numerous trials. The mechanism of these viruses involves both direct tumor cell lysis and the induction of immunogenic cell death icd. Methods and protocols, methods in molecular biology, vol. Ex vivo infection of live tissue with oncolytic viruses. Get your kindle here, or download a free kindle reading app. Pioneering work done by lindenmann and klein in 1967 demonstrated that viral oncolysis of tumor cells by influenza virus increases immunogenicity of tumor cell antigens. In addition, given major differences in the biology of popular oncolytic viruses, there might be different optimal uses for each. Genetic engineering of oncolytic viruses for cancer therapy. Ovs often induce immunogenic cell death icd of cancer cells, and they may interact directly with immune cells as well to prime antitumor immunity.
Clinical trials have explored a wide variety of viruses including naturally occurring viruses and genetically engineered viruses. Oncolytic virus mg1 targets and eliminates cells latently. Oncolytic viruses replicate in tumor cells and induce a cytopathic effect or kill tumor cells through other means, such as induction of apoptosis. The industrialscale manufacturing of viruses or virus like particles in cell culture is necessary for gene therapy and the treatment of cancer with oncolytic viruses. The first commercialized oncolytic virus product in the world. Differential equation models frequently paired with experimental data have been successfully used to improve treatment protocols involving oncolytic viruses 1116. Recently, oncolytic virotherapy has shown promising results in treating hccs and the effects can be more enhanced by adopting immune modulatory molecules. Written in the highly successful methods in molecular biology series format. This protocol describes how to process tissues for ex vivo infection with oncolytic virus and subsequent viral quantification. They are genetically engineered so that they can replicate in cancer cells but not in normal cells, and are used in oncolytic virotherapy. As the infected cancer cells are destroyed by oncolysis, they release new infectious virus particles or virions to help destroy the remaining tumour. Oncolytic virus oncolytic viruses ovs are a class of viruses that can selectively replicate in and lyse cancer cells, and subsequent spread with a tumor while not causing damage to normal cells.
Development of a versatile oncolytic virus platform for local. Since the first report of an engineered oncolytic virus, there has been a continuing and steady. The clearance of tumor depends on the balance between antiviral and antitumor immunity. Wo2016185414a1 recombinant oncolytic viruses and uses. This diversity of virotherapeutics not only offers interesting opportunities for the. Oncolytic viruses as therapeutic cancer vaccines molecular. As such, it comprises a collection of 15 chapters that span a little less than 250 pages. The ultimate goal was to develop svv001 as an oncolytic virus for the treatment of metastatic cancers that have neuroendocrine properties. Armed oncolytic viruses seek to complement the potency of the oncolytic virus by the addition of therapeutic transgenes.
1061 413 34 175 1056 274 316 1058 1477 1586 1156 460 1595 154 718 764 491 1330 1111 557 820 740 1 271 1247 862 1207 638 239 807 306 1074 774 1419 818 796 1352 740 751 155 192