• NK cell
• T cell
• autophagy
• cancer immunotherapy
• cancer therapy resistance
• cancer vaccine
• dendritic cell
• exosome
• immunogenic cell death
• interferon-Ι

• biochemistry
• biological techniques
• biotechnology
• cancer
• genetics
• molecular biology
• oncology

As an oncolytic virus, Newcastle disease virus (NDV) can specifically kill tumor cells and has been tested as an attractive oncolytic agent for cancer virotherapy. Virus infection can trigger the changes of the cellular microRNA (miRNA) expression profile, which can greatly influence viral replication and pathogenesis. However, the interplay between NDV replication and cellular miRNA expression in tumor cells is still largely unknown. In the present study, we compared the profiles of cellular miRNAs in uninfected and NDV-infected HeLa cells by small RNA deep sequencing. Here we report that NDV infection in HeLa cells significantly changed the levels of 40 miRNAs at 6 h post-infection (hpi) and 62 miRNAs at 12 hpi. Among 23 highly differentially expressed miRNAs, NDV infection greatly promoted the levels of 3 miRNAs and suppressed the levels of 20 miRNAs at both time points. These 23 miRNAs are predicted to target various genes involved in virus replication and antiviral immunity such as ErbB, Jak-STAT, NF-kB and RIG-I-like receptor. Verification of deep sequencing results by quantitative RT-PCR showed that 9 out of 10 randomly selected miRNAs chosen from this 23-miRNA pool were consistent with deep sequencing data, including 6 down-regulated and 3 up-regulated. Further functional research revealed that hsa-miR-4521, a constituent in this 23-miRNA pool, inhibited NDV replication in HeLa cells. Moreover, dual-luciferase and gene expression array uncovered that the member A of family with sequence similarity 129 (FAM129A) was directly targeted by hsa-miR-4521 and positively regulated NDV replication in HeLa cells, indicating that hsa-miR-4521 may regulate NDV replication via interaction with FAM129A. To our knowledge, this is the first report of the dynamic cellular miRNA expression profile in tumor cells after NDV infection and may provide a valuable basis for further investigation on the roles of miRNAs in NDV-mediated oncolysis.

• FAM129A
• HeLa cells
• NDV
• hsa-miR-4521
• miRNA expression profile
• oncologic virus

• Oncolytic viruses
• cancer therapy
• clinical trials

Colorectal cancer (CRC) is one of the main reasons of tumor-related deaths worldwide. At present, the main treatment is surgery, but the results are unsatisfactory, and the prognosis is poor. The majority of patients die due to liver or lung metastasis or recurrence. In recent years, great progress has been made in the field of tumor gene therapy, providing a new treatment for combating CRC. As oncolytic viruses selectively replicate almost exclusively in the cytoplasm of tumor cells and do not require integration into the host genome, they are safer, more effective and more attractive as oncolytic agents. Newcastle disease virus (NDV) is a natural RNA oncolytic virus. After NDV selectively infects tumor cells, the immune response induced by NDV’s envelope protein and intracellular factors can effectively kill the tumor without affecting normal cells. Reverse genetic techniques make NDV a vector for gene therapy. Arming the virus by inserting various exogenous genes or using NDV in combination with immunotherapy can also improve the anti-CRC capacity of NDV, and good results have been achieved in animal models and clinical treatment trials. This article reviews the molecular biological characteristics and oncolytic mechanism of NDV and discusses in vitro and in vivo experiments on NDV anti-CRC capacity and clinical treatment. In conclusion, NDV is an excellent candidate for cancer treatment, but more preclinical studies and clinical trials are needed to ensure its safety and efficacy.

• Newcastle disease virus
• Exogenous gene
• Armed virus
• Oncolytic therapy
• Colorectal cancer

• directed viral evolution
• oncolytic viruses
• virus adaptation
• virus selection

Background: Oncolytic viruses (OVs) are emerging as potent inducers of immunogenic cell death (ICD), releasing danger-associated molecular patterns (DAMPs) that induce potent anticancer immunity. Oncolytic Newcastle disease virus (NDV) has been shown to educe ICD in both glioma and lung cancer cells. The objective of this study is to investigate whether oncolytic NDV induces ICD in melanoma cells and how it is regulated.

Methods: Various time points were actuated to check the expression and release of ICD markers induced by NDV strain, NDV/FMW in melanoma cell lines. The expression and release of ICD markers induced by oncolytic NDV strain, NDV/FMW, in melanoma cell lines at various time points were determined. Surface-exposed calreticulin (CRT) was inspected by confocal imaging. The supernatants of NDV/FMW infected cells were collected and concentrated for the determination of ATP secretion by ELISA, HMGB1, and HSP70/90 expression by immunoblot (IB) analysis. Pharmacological inhibition of apoptosis, autophagy, necroptosis, ER Stress, and STAT3 (signal transducer and activator of transcription 3) was achieved by treatment with small molecule inhibitors. Melanoma cell lines stably depleted of STAT3 were established with lentiviral constructs. Supernatants from NDV-infected cells were intratumorally injected to mice bearing melanoma cells-derived tumors.

Results: Oncolytic NDV induced CRT exposure, the release of HMGB1 and HSP70/90 as well as secretion of ATP in melanoma cells. Inhibition of apoptosis, autophagy, necroptosis or ER stress attenuated NDV/FMW-induced release of HMGB1 and HSP70/90. Moreover, NDV/FMW-induced ICD markers in melanoma cells were also suppressed by either treatment with a STAT3 inhibitor or shRNA-mediated depletion of STAT3. Of translational importance, treatment of mice bearing melanoma cells-derived tumors with supernatants from NDV/FMW-infected cells significantly inhibited tumor growth.

Conclusions: Our data authenticate that oncolytic NDV/FMW might be a potent inducer of ICD in melanoma cells, which is amalgamated with several forms of cell death. We also show that STAT3 plays a role in NDV/FMW-induced ICD in melanoma cells. Together, our data highlight oncolytic NDV as propitious for cancer therapeutics by stimulatingan anti-melanoma immune response.

• immunogenic cell death
• melanoma
• newcastle disease virus
• signal transducer and activator of transcription 3
• virotherapy

• Oncolytic virus
• antitumor immune response
• herpes simplex virus
• immunotherapy
• newcastle disease virus
• oncolysis
• productive infection
• reovirus
• vaccinia virus
• vesicular stomatitis virus

Oncolytic viruses are under intense development and have earned their place among the novel class of cancer immunotherapeutics that are changing the face of cancer therapy. Their ability to specifically infect and efficiently kill tumor cells, while breaking immune tolerance and mediating immune responses directed against the tumor, make oncolytic viruses highly attractive candidates for immunotherapy. Increasing evidence indicates that a subclass of oncolytic viruses, which encodes for fusion proteins, could outperform non-fusogenic viruses, both in their direct oncolytic potential, as well as their immune-stimulatory properties. Tumor cell infection with these viruses leads to characteristic syncytia formation and cell death due to fusion, as infected cells become fused with neighboring cells, which promotes intratumoral spread of the infection and releases additional immunogenic signals. In this review, we discuss the potential of fusogenic oncolytic viruses as optimal candidates to enhance immunotherapy and initiate broad antitumor responses. We provide an overview of the cytopathic mechanism of syncytia formation through viral-mediated expression of fusion proteins, either endogenous or engineered, and their benefits for cancer therapy. Growing evidence indicates that fusogenicity could be an important feature to consider in the design of optimal oncolytic virus platforms for combinatorial oncolytic immunotherapy.

• cancer
• fusion
• fusogenic
• fusogenicity
• immunogenic
• immunotherapy
• oncolytic
• syncytium
• virus

To investigate the specific role of immune responses induced by lentogenic Newcastle disease virus (NDV) for its antitumor effect.

NDV LaSota strain was used to infect the following human cells: non-small cell lung carcinoma (A549), glioblastoma (U87MG and T98G), mammary gland adenocarcinoma (MCF7 and MDA-MB-453), hepatocellular carcinoma (Huh7), transformed embryonic kidney cells (HEK293), primary monocytes, lung fibroblast (HF19), skin fibroblast (NB1RGB) and rat astroglia (RCR-1) at 0.001 multiplicity of infection. NDV-induced cytotoxicity and expression of proinflammatory cytokines were analyzed using 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide assay and multiplex enzyme-linked immunosorbent assay, respectively.

Tumor cells (A549, U87MG, T98G, Huh7, MDA-MB-453, and MCF7) showed viability of <44%, while normal cell lines HEK293, NB1RGB, and RCR-1 showed 84%, 73%, and 69% viability at 72 hours postinfection, respectively. Proinflammatory cytokine profiling showed that NDV mainly induced the secretion of interferon (IFN)-α, IFN-β, and IFN-λ in tumor cells and only IFN-λ in normal cells. In addition, NDV infection induced the production of interleukin (IL)-6 in most cells.

Our findings suggest a new perspective regarding the role of IFN-λ and IL-6 in the mechanism of tumor selectivity and oncolysis of NDV.

• Newcastle disease virus
• cytokines
• interferons
• oncolytic virus

• 5-FU (5-fluorouracil)
• NDV (Newcastle disease virus)
• virotherapy

Preclinical studies demonstrate that a broad spectrum of human malignant cells can be killed by oncolytic paramyxoviruses, which include cells of ecto-, endo-, and mesodermal origin. In clinical trials, significant reduction in size or even complete elimination of primary tumors and established metastases are reported. Different routes of viral administration (intratumoral, intravenous, intradermal, intraperitoneal, or intrapleural), and single- versus multiple-dose administration schemes have been explored. The reported side effects are grade 1 and 2, with the most common among them being mild fever. Some advantages in using paramyxoviruses as oncolytic agents versus representatives of other viral families exist. The cytoplasmic replication results in a lack of host genome integration and recombination, which makes paramyxoviruses safer and more attractive candidates for widely used therapeutic oncolysis in comparison with retroviruses or some DNA viruses. The list of oncolytic paramyxovirus representatives includes attenuated measles virus (MV), mumps virus (MuV), low pathogenic Newcastle disease (NDV), and Sendai (SeV) viruses. Metastatic cancer cells frequently overexpress on their surface some molecules that can serve as receptors for MV, MuV, NDV, and SeV. This promotes specific viral attachment to the malignant cell, which is frequently followed by specific viral replication. The paramyxoviruses are capable of inducing efficient syncytium-mediated lyses of cancer cells and elicit strong immunomodulatory effects that dramatically enforce anticancer immune surveillance. In general, preclinical studies and phase 1–3 clinical trials yield very encouraging results and warrant continued research of oncolytic paramyxoviruses as a particularly valuable addition to the existing panel of cancer-fighting approaches.

• Animals
• Cancer Vaccines/genetics
• Cancer Vaccines/immunology
• Genetic Vectors/genetics
• Genetic Vectors/immunology
• Humans
• Immunologic Factors
• Immunotherapy
• Immunotherapy, Adoptive
• Neoplasms/genetics
• Neoplasms/immunology
• Neoplasms/therapy
• Oncolytic Virotherapy
• Oncolytic Viruses/genetics
• Oncolytic Viruses/immunology

• Canadian Institutes of Health Research

• Newcastle disease virus
• Paramyxovirus
• Pathogenicity
• Reverse genetics system

Oncolytic virotherapy has been an attractive drug platform for targeted therapy of cancer over the past few years. Viral vectors can be used to target and lyse cancer cells, but achieving good efficacy and specificity with this treatment approach is a major challenge. Here, we assessed the ability of a novel dual-specific anti-tumor oncolytic adenovirus, expressing the hemagglutinin-neuraminidase (HN) gene from the Newcastle disease virus under the human telomerase reverse transcriptase (hTERT) promoter (Ad-hTERTp-E1a-HN), to inhibit esophageal cancer EC-109 cells in culture and to reduce tumor burden in xenografted BALB/c nude mice. In vitro, infection with Ad-hTERT-E1a-HN could inhibit the growth of EC-109 cells significantly and also protect normal human liver cell line L02 from growth suppression in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Ad-hTERT-E1a-HN also effectively and selectively decreased the sialic acid level on EC-109 cells, but not on L02 cells. Furthermore, Ad-hTERT-E1a-HN was shown to induce the apoptosis pathway via acridine orange and ethidium bromide staining (AO/EB staining), increase reactive oxygen species (ROS), reduce mitochondrial membrane potential and release cytochrome c. In vivo, xenografted BALB/c nude mice were treated via intratumoral or intravenous injections of Ad-hTERT-E1a-HN. Although both treatments showed an obvious suppression in tumor volume, only Ad-hTERT-E1a-HN delivered via intratumoral injection elicited a complete response to treatment. These results reinforced previous findings and highlighted the potential therapeutic application of Ad-hTERT-E1a-HN for treatment of esophageal cancer in clinical trials.

• Adenoviruses, Human/genetics
• Adenoviruses, Human/growth & development
• Animals
• Cell Line, Tumor
• Disease Models, Animal
• Female
• Gene Expression
• HN Protein/genetics
• Humans
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Neoplasms/therapy
• Newcastle disease virus/genetics
• Oncolytic Virotherapy/methods
• Oncolytic Viruses/genetics
• Oncolytic Viruses/growth & development
• Treatment Outcome

• cancer
• gene therapy
• oncolytic therapy
• treatment
• virus

Newcastle disease virus (NDV) is an avian virus that causes deadly infection to over 250 species of birds, including domestic and wild-type, thus resulting in substantial losses to the poultry industry worldwide. Many reports have demonstrated the oncolytic effect of NDV towards human tumor cells. The interesting aspect of NDV is its ability to selectively replicate in cancer cells. Some of the studies have undergone human clinical trials, and favorable results were obtained. Therefore, NDV strains can be the potential therapeutic agent in cancer therapy. However, investigation on the therapeutic perspectives of NDV, especially human immunological effects, is still ongoing. This paper provides an overview of the current studies on the cytotoxic and anticancer effect of NDV via direct oncolysis effects or immune stimulation. Safety of NDV strains applied for cancer immunotherapy is also discussed in this paper.

Newcastle disease virus (NDV) is an avian virus that is attenuated in primates and is a potential vaccine vector for human use. We evaluated NDV as a vector for expressing selected antigens of the Lyme disease pathogen Borrelia burgdorferi. A series of recombinant NDVs were generated that expressed intracellular or extracellular forms of two B. burgdorferi antigens: namely, the basic membrane protein A (BmpA) and the outer surface protein C (OspC). Expression of the intracellular and extracellular forms of these antigens was confirmed in cultured chicken cells. C3H or Balb/C mice that were immunized intranasally with the NDV vectors mounted vigorous serum antibody responses against the NDV vector, but failed to mount a robust response against either the intracellular or extracellular forms of BmpA or OspC. By contrast, a single immunization of hamsters with the NDV vectors via the intranasal, intramuscular, or intraperitoneal route resulted in rapid and rigorous antibody responses against the intracellular or extracellular forms of BmpA and OspC. When groups of hamsters were separately inoculated with various NDV vectors and challenged with B. burgdorferi (10⁸ cells/animal), immunization with vector expressing either intracellular or extracellular BmpA was associated with a significant reduction of the pathogen load in the joints. Taken together, our studies highlighted the importance of NDV as vaccine vector that can be used for simple yet effective immunization of hosts against bacterial infections including Lyme disease.

• oncolytic viral therapy
• malignant glioma
• g207
• hsv 1716
• onyx-015 adenovirus
• reolysin
• vaccinia
• newcastle disease virus
• measles virus

• Adenoviridae/genetics
• Adenoviridae/physiology
• Animals
• Brain Neoplasms/genetics
• Brain Neoplasms/therapy
• Clinical Trials as Topic
• Genetic Vectors
• Glioma/genetics
• Glioma/therapy
• Herpesvirus 1, Human/genetics
• Herpesvirus 1, Human/physiology
• Humans
• Oncolytic Virotherapy
• Oncolytic Viruses/genetics
• Oncolytic Viruses/physiology
• Paramyxoviridae/genetics
• Paramyxoviridae/physiology
• Poliovirus/genetics
• Poliovirus/physiology
• Poxviridae/genetics
• Poxviridae/physiology
• Reoviridae/genetics
• Reoviridae/physiology
• Rhabdoviridae/genetics
• Rhabdoviridae/physiology

• P01 CA071933 NCI NIH HHS
• P20 CA151129 NCI NIH HHS
• P50 CA097247 NCI NIH HHS

Oncolytic virotherapy is a promising form of gene therapy for cancer, employing nature’s own agents to find and destroy malignant cells. The purpose of this review is to provide an introduction to this very topical field of research and to point out some of the current observations, insights and ideas circulating in the literature. We have strived to acknowledge as many different oncolytic viruses as possible to give a broader picture of targeting cancer using viruses. Some of the newest additions to the panel of oncolytic viruses include the avian adenovirus, foamy virus, myxoma virus, yaba-like disease virus, echovirus type 1, bovine herpesvirus 4, Saimiri virus, feline panleukopenia virus, Sendai virus and the non-human coronaviruses. Although promising, virotherapy still faces many obstacles that need to be addressed, including the emergence of virus-resistant tumor cells.

• Animals
• Disease Models, Animal
• Genetic Therapy/methods
• Humans
• Neoplasms/pathology
• Neoplasms/therapy
• Neoplasms/virology
• Oncolytic Virotherapy/methods
• Oncolytic Viruses/classification
• Oncolytic Viruses/physiology
• Virus Replication

• Animals
• Cell Line
• Gene Expression Regulation, Viral
• Interferons/biosynthesis
• Interferons/immunology
• Newcastle Disease/physiopathology
• Newcastle disease virus/physiology
• Tumor Cells, Cultured/metabolism
• Tumor Cells, Cultured/virology
• Viral Proteins/genetics
• Viral Proteins/physiology

• R01 AI038956 NIAID NIH HHS
• R01 AI055940 NIAID NIH HHS
• AI 38956 NIAID NIH HHS
• AI 055940 NIAID NIH HHS

AIM: To treat patients with stage I-IV malignant tumors of digestive tract using autologous tumor cell vaccine and NDV (Newcastle disease virus) vaccine, and observe the survival period and curative effect.

METHODS: 335 patients with malignant tumors of digestive tract were treated with autologous tumor cell vaccine and NDV vaccine. The autologous tumor cell vaccine received were assigned for long-term survival observation. While these failed to obtain the autologous tumor tissue were given with NDV vaccine for a received short-term observation on curative effect.

RESULTS: The colorectal cancer patients treated with autologous tumor cell vaccine were divided into two groups: the controlled group (subjected to resection alone) (n = 257), the vaccine group (subjected to both resection and immunotherapy) (n = 310). 25 patients treated with NDV immunotherapy were all at stage IV without having resection. In postoperation adjuvant therapy patients, the 5, 6 and 7-year survival rates were 66.51%, 60.52%, 56.50% respectively; whereas in patients with resection alone, only 45.57%, 44.76% and 43.42% respectively. The average survival period was 5.13 years (resection alone group 4.15 years), the median survival period was over 7 years (resection alone group 4.46 years). There were significant differences between the two groups. The patients treated with resection plus vaccine were measured delayed-type hypersensitivity (DTH) reactions after vaccination, (indurative scope > 5 mm). The magnitude of DTH was related to the prognosis. The 5-year survival rate was 80% for those with indurations greater than 5 mm, compared with 30% for those with indurations less than 5 mm. The 1-year survival rate was 96% for 25 patients treated with NDV immunotherapy. The total effective rate (CR+PR) was 24.00% in NDV immunotherapy; complete remission (CR) in 1 case (4.00%), partial remission (PR) in 5 cases (20.00%), stabilizedin in 16 cases (64.00%), progression (PD) in 1 case (4.00%). After NDV vaccine immunotherapy, the number of NK cell increased and immune function imporved obviously.

CONCLUSION: The autologous tumor cell vaccine and NDV vaccine can prolong the patients’ life. NDV vaccine is notably effective for short-term with promotion of quality of life and can be used whenever necessary with good prospects.

• Cancer Vaccines/therapeutic use
• Gastrointestinal Neoplasms/therapy
• Humans
• Immunotherapy
• Newcastle disease virus/immunology
• Viral Vaccines/therapeutic use