DOCSLIB.ORG

Synthetic and Systems Biology Principles in the Design of Programmable Oncolytic Virus Immunotherapies for Glioblastoma

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Synthetic and Systems Biology Principles in the Design of Programmable Oncolytic Virus Immunotherapies for Glioblastoma NEUROSURGICAL FOCUS Neurosurg Focus 50 (2):E10, 2021 Synthetic and systems biology principles in the design of programmable oncolytic virus immunotherapies for glioblastoma *Dileep D. Monie, BA,1,6,7 Archis R. Bhandarkar, BS,6 Ian F. Parney, MD, PhD,1,3 Cristina Correia, PhD,5 Jann N. Sarkaria, MD,4 Richard G. Vile, PhD,1,2 and Hu Li, PhD5 Departments of 1Immunology, 2Molecular Medicine, 3Neurosurgery, 4Radiation Oncology, and 5Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic; 6Mayo Clinic Alix School of Medicine; 7Mayo Clinic Graduate School of Biomedical Sciences; and Mayo Clinic College of Medicine and Science, Rochester, Minnesota Oncolytic viruses (OVs) are a class of immunotherapeutic agents with promising preclinical results for the treatment of glioblastoma (GBM) but have shown limited success in recent clinical trials. Advanced bioengineering principles from disciplines such as synthetic and systems biology are needed to overcome the current challenges faced in developing effective OV-based immunotherapies for GBMs, including off-target effects and poor clinical responses. Synthetic biol- ogy is an emerging field that focuses on the development of synthetic DNA constructs that encode networks of genes and proteins (synthetic genetic circuits) to perform novel functions, whereas systems biology is an analytical framework that enables the study of complex interactions between host pathways and these synthetic genetic circuits. In this review, the authors summarize synthetic and systems biology concepts for developing programmable, logic-based OVs to treat GBMs. Programmable OVs can increase selectivity for tumor cells and enhance the local immunological response using synthetic genetic circuits. The authors discuss key principles for developing programmable OV-based immunotherapies, including how to 1) select an appropriate chassis, a vector that carries a synthetic genetic circuit, and 2) design a syn- thetic genetic circuit that can be programmed to sense key signals in the GBM microenvironment and trigger release of a therapeutic payload. To illustrate these principles, some original laboratory data are included, highlighting the need for systems biology studies, as well as some preliminary network analyses in preparation for synthetic biology applications. Examples from the literature of state-of-the-art synthetic genetic circuits that can be packaged into leading candidate OV chassis are also surveyed and discussed. https://thejns.org/doi/abs/10.3171/2020.12.FOCUS20855 KEYWORDS synthetic biology; systems biology; oncolytic virus; cancer immunotherapy; glioblastoma LIOBLASTOMA (GBM) is the most common and tions for many cancer patients but, despite promising safe- lethal malignancy of the CNS. This devastating ty profiles, are not currently effective for those suffering disease has a median overall survival of 3 months from GBM.4 Gfrom the time of diagnosis in untreated patients.1 Despite Oncolytic viruses (OVs) are a class of immunotherapeu- the significant cost and morbidity associated with the stan- tic agents with an FDA-approved treatment for melanoma, dard of care, i.e., resection combined with adjuvant che- a solid tumor.5 OVs work by directly lysing tumor cells, motherapy and radiation, the median patient life expec- which then release tumor antigens in the context of danger tancy is only extended to about 14 months and the disease signals—both damage-associated (DAMPs) and pathogen- remains almost uniformly fatal.2,3 Yet, this is the best we associated (PAMPs) molecular patterns—that elicit antitu- can do for patients after hundreds of clinical trials over mor immunity (Fig. 1A).6 Since 1991, many attempts to use several decades. Immunotherapies are breakthrough op- OVs in GBM have had limited success. More than 20 clini- ABBREVIATIONS Ad5 = adenovirus serotype 5; CCN1 = cellular communication network factor 1; DAMP = damage-associated molecular pattern; dsDNA = double- stranded DNA; GBM = glioblastoma; GFAP = glial fibrillary acidic protein; GM-CSF = granulocyte-macrophage colony-stimulating factor; HSV-1 = herpes simplex virus type 1; hTERT = telomerase reverse transcriptase; IL = interleukin; miRNA = micro RNA; OV = oncolytic virus; PAMP = pathogen-associated molecular pattern; pCancer = cancer-selective promoter; PD-1 = programmed death–1; PD-L1 = programmed death–ligand 1; PDX = patient-derived xenograft; PPI = protein-protein interaction; scFv = single-chain variable fragment; sTRAIL = secreted tumor necrosis factor–related apoptosis-inducing ligand; VSV = vesicular stomatitis virus. SUBMITTED October 1, 2020. ACCEPTED December 4, 2020. INCLUDE WHEN CITING DOI: 10.3171/2020.12.FOCUS20855. * D.D.M. and A.R.B. contributed equally to this work. ©AANS 2021, except where prohibited by US copyright law Neurosurg Focus Volume 50 • February 2021 1 Unauthenticated | Downloaded 10/10/21 07:15 PM UTC Monie et al. FIG. 1. A: Mechanisms of OV immunotherapy for GBM. B: Sense-compute-actuate framework of programmable OV immunothera- py for GBM. C: Design-build-test-analyze development cycle of synthetic OVs for GBM blends systems and synthetic biology. 2 Neurosurg Focus Volume 50 • February 2021 Unauthenticated | Downloaded 10/10/21 07:15 PM UTC Monie et al. cal trials of 7 different OVs did not translate encouraging tropisms. To generate antitumor immunity, the innate im- preclinical results to patients with GBM.7 The root causes mune system must be activated at some point during OV of these failures can be traced to failed oncotropism, de- replication. This awakens the adaptive immune system spite neurosurgical delivery, and oncolysis mechanisms.7 to precisely target all cancerous cells, both infected and Therefore, advanced bioengineering methods are neces- uninfected.15 If the patient has been exposed to the viral sary to design and implement better armaments. chassis before, by either infection or vaccination, then this Synthetic biology involves the creation and manipula- may limit the efficacy of the OV. Host genome integra- tion of biological systems using rational, modular design tion (e.g., lentivirus) may be advantageous for persistent principles from electrical engineering, computer sci- antitumor effects but comes with the risk of genetic dam- ence, and related disciplines.8 The sense-compute-actuate age and chronic latent infection.16 Ultimately, however, the framework (Fig. 1B)9 is a guiding design principle in syn- immune system must be able to clear the OV infection, thetic biology, in which long stretches of synthetic DNA particularly in the CNS, to prevent chronic inflammation constructs that encode networks of genes and proteins and its sequelae. (synthetic genetic circuits) are designed to sense stimuli of interest, compute which environmental state a cell is in Neurotropic Viruses based on a permutation of stimuli, and actuate a response A chassis needs to target glioma cells, which arise from accordingly. In November 2019, the first living medicine healthy cells of the brain parenchyma and share many cell containing a synthetic genetic circuit entered phase 1 surface and metabolic features, so naturally occurring clinical trials in the form of a bacteria-based cancer im- neurotropic viruses are the obvious starting point. munotherapy.10 Such engineered genetic circuits are now primed to be packaged into OVs and programmed to co- Herpes Simplex Virus Type 1 ordinate local tissue responses, with preclinical studies Herpes simplex virus type 1 (HSV-1) is an enveloped demonstrating enhanced 1) tumor cell targeting and 2) virus with a linear 152-kilobase (Kb) double-stranded generation of antitumor immunity. DNA (dsDNA) genome.17 It is capable of carrying large Systems biology complements synthetic biology by en- payloads—as much as 20 Kb—making it particularly at- abling complex design and analysis of genetic circuits. Tu- tractive for synthetic biology applications. It is the first and mor microenvironment cell states can be defined by using only OV that has garnered FDA approval with talimogene single-cell multiomics data sets such as RNA sequencing laherparepvec, which has been engineered extensively to (RNA-seq) and mass cytometry. These high-dimensional target melanoma.18 Using HSV-1 against GBM was also data sets are amenable to machine learning11 and network- the first proposed OV with experimental success in a mu- based classification.12 Computation identifies biological rine model;19 therefore, HSV-1 is particularly promising as features unique to glioma cells,13 elucidates pathways a treatment for GBM. orthogonal to genetic circuit designs, and helps priori- The high prevalence of HSV-1 infection in humans, tize OV-based approaches.14 After the synthetic OVs are however, means that preexisting immunity (e.g., neutral- constructed and tested, further systems analyses ensure izing antibodies) poses a barrier to its application as an ef- that the OVs have predictable effects on the cells and the fective OV. Modulating this OV-host immune interaction greater tumor microenvironment, statistically accounting is a possibility with synthetic gene circuits.20 Additionally, for complex stochastic behavior. This knowledge is fed several molecular features of GBM have been identified back into the design of improved, next-generation OV ge- that influence the efficacy of HSV-1 OV in vitro and in netic circuits (Fig. 1C). vivo, including the integrin ligand cellular communication network factor 1 (CCN1) protein found in the extracellular Selection and Delivery of OV Chassis matrix
Load more

Recommended publications
  • Promoting the Accumulation of Tumor-Specific T Cells in Tumor
    PROTOCOL Promoting the accumulation of tumor-specific T cells in tumor tissues by dendritic cell vaccines and chemokine-modulating agents Nataša Obermajer1 , Julie Urban2, Eva Wieckowski1,2,6, Ravikumar Muthuswamy1, Roshni Ravindranathan1, David L Bartlett1 & Pawel Kalinski1–5 1Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. 2Immunotransplantation Center, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. 3University of Pittsburgh Cancer Institute, Hillman Cancer Center, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. 4Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. 5Department of Bioengineering, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. 6Deceased. Correspondence should be addressed to P.K. ([email protected]). Published online 18 January 2018; doi:10.1038/nprot.2017.130 This protocol describes how to induce large numbers of tumor-specific cytotoxic T cells (CTLs) in the spleens and lymph nodes of mice receiving dendritic cell (DC) vaccines and how to modulate tumor microenvironments (TMEs) to ensure effective homing of the vaccination-induced CTLs to tumor tissues. We also describe how to evaluate the numbers of tumor-specific CTLs within tumors. The protocol contains detailed information describing how to generate a specialized DC vaccine with augmented ability to induce tumor-specific CTLs. We also describe methods to modulate the production of chemokines in the TME and show how to quantify tumor-specific CTLs in the lymphoid organs and tumor tissues of mice receiving different treatments. The combined experimental procedure, including tumor implantation, DC vaccine generation, chemokine-modulating (CKM) approaches, and the analyses of tumor-specific systemic and intratumoral immunity is performed over 30–40 d. The presented ELISpot-based ex vivo CTL assay takes 6 h to set up and 5 h to develop.
    [Show full text]
  • Pharmacological Improvement of Oncolytic Virotherapy
    PHARMACOLOGICAL IMPROVEMENT OF ONCOLYTIC VIROTHERAPY Mohammed Selman Thesis submitted to the Faculty of Graduate and Postdoctoral Studies in partial fulfillment of the requirements for the Doctorate of Philosophy in Biochemistry Department of Biochemistry, Microbiology & Immunology Faculty of Medicine University of Ottawa © Mohammed Selman, Ottawa, Canada, 2018 Abstract Oncolytic viruses (OV) are an emerging class of anticancer bio-therapeutics that induce antitumor immunity through selective replication in cancer cells. However, the efficacy of OVs as single agents remains limited. We postulate that resistance to oncolytic virotherapy results in part from the failure of tumor cells to be sufficiently infected. In this study, we provide evidence that in the context of sarcoma, a highly heterogeneous malignancy, the infection of tumors by different oncolytic viruses varies greatly. Similarly, for a given oncolytic virus, productive infection of tumors across patient samples varies by many orders of magnitude. To overcome this issue, we hypothesize that the infection of resistant tumors can be achieved through the use of selected small molecules. Here, we have identified two novel drug classes with the ability to improve the efficacy of OV therapy: fumaric and maleic acid esters (FMAEs) and vanadium compounds. FMAEs are enhancing infection of cancer cells by several oncolytic viruses in cancer cell lines and human tumor biopsies. The ability of FMAEs to enhance viral spread is due to their ability to inhibit type I IFN production and response, which is associated with their ability to block nuclear translocation of transcription factor NF-κB. Vanadium-based phosphatase inhibitors enhance OV infection of RNA viruses in vitro and ex vivo, in resistant cancer cell lines.
    [Show full text]
  • Oncolytic Herpes Simplex Virus-Based Therapies for Cancer
    cells Review Oncolytic Herpes Simplex Virus-Based Therapies for Cancer Norah Aldrak 1 , Sarah Alsaab 1,2, Aliyah Algethami 1, Deepak Bhere 3,4,5 , Hiroaki Wakimoto 3,4,5,6 , Khalid Shah 3,4,5,7, Mohammad N. Alomary 1,2,* and Nada Zaidan 1,2,* 1 Center of Excellence for Biomedicine, Joint Centers of Excellence Program, King Abdulaziz City for Science and Technology, P.O. Box 6086, Riyadh 11451, Saudi Arabia; [email protected] (N.A.); [email protected] (S.A.); [email protected] (A.A.) 2 National Center for Biotechnology, Life Science and Environmental Research Institute, King Abdulaziz City for Science and Technology, P.O. Box 6086, Riyadh 11451, Saudi Arabia 3 Center for Stem Cell Therapeutics and Imaging (CSTI), Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA; [email protected] (D.B.); [email protected] (H.W.); [email protected] (K.S.) 4 Department of Neurosurgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA 5 BWH Center of Excellence for Biomedicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA 6 Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA 7 Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA * Correspondence: [email protected] (M.N.A.); [email protected] (N.Z.) Abstract: With the increased worldwide burden of cancer, including aggressive and resistant cancers, oncolytic virotherapy has emerged as a viable therapeutic option. Oncolytic herpes simplex virus (oHSV) can be genetically engineered to target cancer cells while sparing normal cells.
    [Show full text]
  • IL-6 Signaling Blockade During CD40-Mediated Immune Activation Favors Antitumor Factors by Reducing TGF-Β, Collagen Type I
    IL-6 Signaling Blockade during CD40-Mediated Immune Activation Favors Antitumor Factors by Reducing TGF-β, Collagen Type I, and PD-L1/PD-1 This information is current as of September 26, 2021. Emma Eriksson, Ioanna Milenova, Jessica Wenthe, Rafael Moreno, Ramon Alemany and Angelica Loskog J Immunol 2019; 202:787-798; Prepublished online 7 January 2019; doi: 10.4049/jimmunol.1800717 Downloaded from http://www.jimmunol.org/content/202/3/787 References This article cites 50 articles, 12 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/202/3/787.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 26, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2019 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology IL-6 Signaling Blockade during CD40-Mediated Immune Activation Favors Antitumor Factors by Reducing TGF-b, Collagen Type I, and PD-L1/PD-1 Emma Eriksson,* Ioanna Milenova,* Jessica Wenthe,* Rafael Moreno,† Ramon Alemany,† and Angelica Loskog*,‡ IL-6 plays a role in cancer pathogenesis via its connection to proteins involved in the formation of desmoplastic stroma and to immunosuppression by driving differentiation of myeloid suppressor cells together with TGF-b.
    [Show full text]
  • Development of a New Hyaluronic Acid Based Redox-Responsive Nanohydrogel for the Encapsulation of Oncolytic Viruses for Cancer Immunotherapy
    nanomaterials Article Development of a New Hyaluronic Acid Based Redox-Responsive Nanohydrogel for the Encapsulation of Oncolytic Viruses for Cancer Immunotherapy Siyuan Deng 1, Alessandra Iscaro 2, Giorgia Zambito 3,4, Yimin Mijiti 5 , Marco Minicucci 5 , Magnus Essand 6, Clemens Lowik 3,4 , Munitta Muthana 2, Roberta Censi 1, Laura Mezzanotte 3,4 and Piera Di Martino 1,* 1 School of Pharmacy, University of Camerino, Via S. Agostino 1, 62032 Camerino, Italy; [email protected] (S.D.); [email protected] (R.C.) 2 Medical School, University of Sheffield, Beech Hill Road, Sheffield S10 2RX, UK; [email protected] (A.I.); m.muthana@sheffield.ac.uk (M.M.) 3 Department of Radiology and Nuclear Medicine, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands; [email protected] (G.Z.); [email protected] (C.L.); [email protected] (L.M.) 4 Department of Molecular Genetics, Erasmus Medical Center, 3015 GD Rotterdam, The Netherlands 5 Physics Division, School of Science and Technology, University of Camerino, Via Madonna delle Carceri 9, 62032 Camerino, Italy; [email protected] (Y.M.); [email protected] (M.M.) 6 Department of Immunology, Genetics and Pathology, Uppsala University, SE-751 85 Uppsala, Sweden; [email protected] * Correspondence: [email protected]; Tel.: +39-0737-40-2215 Abstract: Oncolytic viruses (OVs) are emerging as promising and potential anti-cancer thera- peutic agents, not only able to kill cancer cells directly by selective intracellular viral replication, Citation: Deng, S.; Iscaro, A.; but also to promote an immune response against tumor. Unfortunately, the bioavailability Zambito, G.; Mijiti, Y.; Minicucci, M.; under systemic administration of OVs is limited because of undesired inactivation caused by Essand, M.; Lowik, C.; Muthana, M.; host immune system and neutralizing antibodies in the bloodstream.
    [Show full text]
  • Treating Cancerous Cells with Viruses: Insights from a Minimal Model for Oncolytic Virotherapy
    LETTERS IN BIOMATHEMATICS 2018, VOL. 5, NO. S1, S117–S136 https://doi.org/10.1080/23737867.2018.1440977 RESEARCH ARTICLE OPEN ACCESS Treating cancerous cells with viruses: insights from a minimal model for oncolytic virotherapy Adrianne L. Jennera, Adelle C. F. Costerb, Peter S. Kima and Federico Frascolic aSchool of Mathematics and Statistics, University of Sydney, Sydney, Australia; bSchool of Mathematics and Statistics, University of New South Wales, Sydney, Australia; cFaculty of Science, Engineering and Technology, Department of Mathematics, Swinburne University of Technology, Melbourne, Australia ABSTRACT ARTICLE HISTORY In recent years, interest in the capability of virus particles as a Received 13 December 2017 treatment for cancer has increased. In this work, we present a Accepted 30 January 2018 mathematical model embodying the interaction between tumour KEYWORDS cells and virus particles engineered to infect and destroy cancerous Oncolytic virotherapy; tissue. To quantify the effectiveness of oncolytic virotherapy, we bifurcation analysis; conduct a local stability analysis and bifurcation analysis of our model. oscillations In the absence of tumour growth or viral decay, the model predicts that oncolytic virotherapy will successfully eliminate the tumour cell population for a large proportion of initial conditions. In comparison, for growing tumours and decaying viral particles there are no stable equilibria in the model; however, oscillations emerge for certain regions in our parameter space. We investigate how the period and amplitude of oscillations depend on tumour growth and viral decay. We find that higher tumour replication rates result in longer periods between oscillations and lower amplitudes for uninfected tumour cells. From our analysis, we conclude that oncolytic viruses can reduce growing tumours into a stable oscillatory state, but are insufficient to completely eradicate them.
    [Show full text]
  • Combination Therapy of Novel Oncolytic Adenovirus with Anti-PD1 Resulted in Enhanced Anti-Cancer Effect in Syngeneic Immunocompetent Melanoma Mouse Model
    pharmaceutics Article Combination Therapy of Novel Oncolytic Adenovirus with Anti-PD1 Resulted in Enhanced Anti-Cancer Effect in Syngeneic Immunocompetent Melanoma Mouse Model Mariangela Garofalo 1,* , Laura Bertinato 1, Monika Staniszewska 2, Magdalena Wieczorek 3 , Stefano Salmaso 1 , Silke Schrom 4, Beate Rinner 4, Katarzyna Wanda Pancer 3 and Lukasz Kuryk 3,5,* 1 Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy; [email protected] (L.B.); [email protected] (S.S.) 2 Centre for Advanced Materials and Technologies, Warsaw University of Technology, Poleczki 19, 02-822 Warsaw, Poland; [email protected] 3 Department of Virology, National Institute of Public Health—National Institute of Hygiene, Chocimska 24, 00-791 Warsaw, Poland; [email protected] (M.W.); [email protected] (K.W.P.) 4 Division of Biomedical Research, Medical University of Graz, Roseggerweg 48, 8036 Graz, Austria; [email protected] (S.S.); [email protected] (B.R.) 5 Clinical Science, Targovax Oy, Lars Sonckin kaari 14, 02600 Espoo, Finland * Correspondence: [email protected] (M.G.); [email protected] (L.K.) Abstract: Malignant melanoma, an aggressive form of skin cancer, has a low five-year survival rate Citation: Garofalo, M.; Bertinato, L.; in patients with advanced disease. Immunotherapy represents a promising approach to improve sur- Staniszewska, M.; Wieczorek, M.; vival rates among patients at advanced stage. Herein, the aim of the study was to design and produce, Salmaso, S.; Schrom, S.; Rinner, B.; by using engineering tools, a novel oncolytic adenovirus AdV-D24- inducible co-stimulator ligand Pancer, K.W.; Kuryk, L.
    [Show full text]
  • Review the Oncolytic Virotherapy Treatment Platform for Cancer
    Cancer Gene Therapy (2002) 9, 1062 – 1067 D 2002 Nature Publishing Group All rights reserved 0929-1903/02 $25.00 www.nature.com/cgt Review The oncolytic virotherapy treatment platform for cancer: Unique biological and biosafety points to consider Richard Vile,1 Dale Ando,2 and David Kirn3,4 1Molecular Medicine Program, Mayo Clinic, Rochester, Minnesota, USA; 2Cell Genesys Corp., Foster City, California, USA; 3Department of Pharmacology, Oxford University Medical School, Oxford, UK; and 4Kirn Oncology Consulting, San Francisco, California, USA. The field of replication-selective oncolytic viruses (virotherapy) has exploded over the last 10 years. As with many novel therapeutic approaches, initial overexuberance has been tempered by clinical trial results with first-generation agents. Although a number of significant hurdles to this approach have now been identified, novel solutions have been proposed and improvements are being made at a furious rate. This article seeks to initiate a discussion of these hurdles, approaches to overcome them, and unique safety and regulatory issues to consider. Cancer Gene Therapy (2002) 9, 1062 – 1067 doi:10.1038/sj.cgt.7700548 Keywords: oncolytic; virotherapy; experimental therapeutics; cancer ew cancer treatments are needed. These agents must limitation of levels of delivery to cancer cells in a solid tumor Nhave novel mechanisms of action and thereby lack mass. Over 10 different virotherapy agents have entered, or cross-resistance with currently available treatments. Viruses will soon be entering, clinical trials; one such adenovirus have evolved to infect, replicate in, and kill human cells (dl1520) has entered a Phase III clinical trial in recurrent through diverse mechanisms. Clinicians treated hundreds of head and neck carcinoma.
    [Show full text]
  • Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus
    viruses Article Forced Degradation Studies to Identify Critical Process Parameters for the Purification of Infectious Measles Virus Daniel Loewe 1,2 , Julian Häussler 1, Tanja A. Grein 1 , Hauke Dieken 1, Tobias Weidner 1, Denise Salzig 1 and Peter Czermak 1,2,3,* 1 Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstraße 14, 35390 Giessen, Germany 2 Faculty of Biology and Chemistry, University of Giessen, Heinrich-Buff-Ring 17, 35392 Giessen, Germany 3 Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Project group Bioresources, Winchesterstr. 3, 35394 Giessen, Germany * Correspondence: [email protected]; Tel.: +49-641-309-2551 Received: 25 June 2019; Accepted: 3 August 2019; Published: 7 August 2019 Abstract: Oncolytic measles virus (MV) is a promising treatment for cancer but titers of up to 1011 infectious particles per dose are needed for therapeutic efficacy, which requires an efficient, robust, and scalable production process. MV is highly sensitive to process conditions, and a substantial fraction of the virus is lost during current purification processes. We therefore conducted forced degradation studies under thermal, pH, chemical, and mechanical stress to determine critical process parameters. We found that MV remained stable following up to five freeze–thaw cycles, but was inactivated during short-term incubation (< 2 h) at temperatures exceeding 35 ◦C. The infectivity of MV declined at pH < 7, but was not influenced by different buffer systems or the ionic strength/osmolality, except high concentrations of CaCl2 and MgSO4. We observed low shear sensitivity (dependent on the flow rate) caused by the use of a peristaltic pump.
    [Show full text]
  • Artificial Riboswitches for Gene Expression and Replication Control of DNA and RNA Viruses
    Artificial riboswitches for gene expression and replication control of DNA and RNA viruses Patrick Ketzera, Johanna K. Kaufmanna,1, Sarah Engelhardta, Sascha Bossowb, Christof von Kalleb, Jörg S. Hartigc, Guy Ungerechtsb,d, and Dirk M. Nettelbecka,2 aOncolytic Adenovirus Group, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany; bDepartment of Translational Oncology, National Center for Tumor Diseases, DKFZ, 69120 Heidelberg, Germany; cDepartment of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, 78457 Konstanz, Germany; and dDepartment of Medical Oncology, National Center for Tumor Diseases, Heidelberg University Hospital, 69120 Heidelberg, Germany Aptazymes are small, ligand-dependent self-cleaving ribozymes virus strains for which pathogenicity is of concern (refs. 2, 3, and that function independently of transcription factors and can be references therein) and of new generations of oncolytic viruses customized for induction by various small molecules. Here, we that were engineered for enhanced potency by different strategies introduce these artificial riboswitches for regulation of DNA and for which side effects in patients cannot be predicted (1, 4). Fur RNA viruses. We hypothesize that they represent universally appli- thermore, transient immunosuppression is being investigated as cable tools for studying viral gene functions and for applications as a means to block antiviral immunity thereby enhancing oncolysis a safety switch for oncolytic and live vaccine viruses. Our study (5). However, this strategy bears an increased risk of uncontrolled shows that the insertion of artificial aptazymes into the adenoviral virus replication, which makes a drug triggered genetic safety immediate early gene E1A enables small-molecule–triggered, dose- switch desirable as a clinical countermeasure.
    [Show full text]
  • Corporate Presentation
    Corporate Presentation April 2020 Forward Looking Safe Harbor Statement This presentation may contain “forward-looking statements” within the meaning of Section 27A of the Securities Act of 1933 and Section 21E of the Securities Exchange Act of 1934, each as amended. These statements are often, but not always, made through the use of words or phrases such as “anticipates”, expects”, plans”, believes”, “intends”, and similar words or phrases. Such statements involve risks and uncertainties that could cause Mustang Bio’s actual results to differ materially from the anticipated results and expectations expressed in these forward-looking statements. These statements are only predictions based on current information and expectations and involve a number of risks and uncertainties. Actual events or results may differ materially from those projected in any such statements due to various factors, including the risks and uncertainties inherent in clinical trials, drug development, and commercialization. You are cautioned not to place undue reliance on these forward-looking statements, which speak only as of the date hereof. All forward-looking statements are qualified in their entirety by this cautionary statement and Mustang Bio undertakes no obligation to update these statements, except as required by law. 2 Building a Fully Integrated Gene & Cell Therapy Company First IND for Mustang-sponsored trial approved; 4 more INDs expected in 2020 ▪ Transformational ex vivo lentiviral gene therapy for XSCID licensed from St. Jude – Promising results in 2 ongoing clinical trials led by St. Jude & NIH ▪ 6 CAR-Ts from COH & FHCRC – all potentially first in class – All programs in Ph 1 trials; 1st Mustang IND to start enrolling 2Q2020 ▪ Ph.
    [Show full text]
  • Combination Immunotherapy Using Oncolytic Virus for the Treatment of Advanced Solid Tumors
    International Journal of Molecular Sciences Review Combination Immunotherapy Using Oncolytic Virus for the Treatment of Advanced Solid Tumors Chang-Myung Oh 1, Hong Jae Chon 2,* and Chan Kim 2,* 1 Department of Biomedical Science and Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Korea; [email protected] 2 Medical Oncology, CHA Bundang Medical Center, CHA University School of Medicine, Seongnam 13497, Korea * Correspondence: [email protected] (H.J.C.); [email protected] (C.K.) Received: 24 September 2020; Accepted: 16 October 2020; Published: 19 October 2020 Abstract: Oncolytic virus (OV) is a new therapeutic strategy for cancer treatment. OVs can selectively infect and destroy cancer cells, and therefore act as an in situ cancer vaccine by releasing tumor-specific antigens. Moreover, they can remodel the tumor microenvironment toward a T cell-inflamed phenotype by stimulating widespread host immune responses against the tumor. Recent evidence suggests several possible applications of OVs against cancer, especially in combination with immune checkpoint inhibitors. In this review, we describe the molecular mechanisms of oncolytic virotherapy and OV-induced immune responses, provide a brief summary of recent preclinical and clinical updates on this rapidly evolving field, and discuss a combinational strategy that is able to overcome the limitations of OV-based monotherapy. Keywords: oncolytic virus; combination immunotherapy; immune checkpoint inhibitor; tumor microenvironment 1. Introduction Viruses are continuously co-evolving with our immune systems and have developed sophisticated mechanisms to manipulate host immune responses [1]. With scientific advances in the understanding of the molecular interplay between viruses and host immune systems, we can develop a new therapeutic modality against cancers using finely tuned viruses, called viroceuticals [2,3].
    [Show full text]