HVJ-Envelope (HVJ-E), an inactivated particle derived from the Hemagglutinating virus of Japan, represents a unique virus-based approach for cancer immunotherapy. Unlike replication-competent oncolytic viruses, which exert antitumor effects through viral propagation and tumor cell lysis, HVJ-E lacks replicative capacity and pathogenicity because its viral genome is fragmented. Conversely, although most non-replicative viral vectors are designed primarily as delivery vehicles, HVJ-E itself possesses intrinsic antitumor activity. By preserving the native viral structure and membrane-fusion capacity of the parental virus, HVJ-E enables fragmented viral genomes to enter target cells, thereby inducing antitumor immune activation and tumor cell death. Mechanistic studies have shown that HVJ-E promotes dendritic cell maturation, activates natural killer cells, and stimulates cytotoxic T lymphocytes, leading to systemic antitumor immunity. Clinical trials have demonstrated favorable safety profiles and encouraging signs of efficacy. Furthermore, reverse translational research has identified combination strategies with T cell co-stimulatory signaling and the apolipoprotein system as a downstream mediator that enhances sensitivity to immune attack. This review summarizes the historical development, mechanistic insights, and emerging therapeutic concepts of HVJ-E as a novel cancer immunotherapy platform. A comprehensive literature search was conducted using PubMed for articles related to HVJ-E and cancer published between 2002 and 2025.

Background Virotherapy eradicates tumors by directly killing cancer cells and causing adjuvant effects. However, the mechanism by which non-replicating virotherapy exerts anti-tumor effects is unclear.Methods In this study, we investigated the genes that mediate the anti-tumor effects of ultraviolet (UV)-irradiated Hemagglutinating Virus of Japan envelope (HVJ-E) using RNA sequencing, gene knockout, and a drug-inducible gene expression system. We examined the antitumor effects of Apolipoprotein d (Apod) using genome-wide CRISPR library screening, in situ biotinylation combined with mass spectrometry, flow cytometry, biochemistry, and tumor-bearing mouse models.Results Here, we show that HVJ-E represses tumor growth via Irf7-induced Apod expression in tumor cells in vivo. Irf7 in B16F10 cells is a pivotal transcription factor for HVJ-E-induced anti-tumor effects. Apod substantially suppresses tumor growth even in HVJ-E-insensitive tumors. Apod is required to increase NKG2D-ligand genes in HVJ-E-treated tumors. Genome-wide CRISPR library screening and in situ biotinylation of Apod reveal an association of Apod with ERK2. Mechanistically, Apod prevents the nuclear translocation of ERK2 and Importin7, increasing NKG2D-ligands expression in B16F10 cells and attenuating tumor growth. Treating a local tumor with a combination therapy of Apod with the anti-OX40, T cell costimulatory molecule, antibody substantially repressed tumor growth in target and non-target lesions alongside T cell activation.Conclusion Our findings provide insights into the molecular mechanisms of how HVJ-E induces anti-tumor effects and can aid the development of therapeutic strategies for eliciting anti-tumor immunity.

Despite advances in androgen receptor signaling inhibitors (ARSIs) and poly (ADP-ribose) polymerase inhibitors (PARPIs), metastatic castration-resistant prostate cancer (mCRPC) remains lethal. PARPIs clinical efficacy is limited in patients with homologous recombination repair deficiencies, such as BRCA1/2 mutations, due to resistance. Thus, identifying novel synthetic lethal interactions with PARP may expand treatment options and improve therapeutic efficacy. Here, to identify genes that influence sensitivity to the PARPI olaparib, we conducted a genome-wide CRISPR-Cas9 knockout screening of 18,010 genes in DU145, 22Rv1, and LNCaP prostate cancer cell lines. Our screening identified PARP and LIG1 as synthetic lethality-inducing factors, whereas TP53 conferred resistance to PARPIs. Simultaneous inhibition of LIG1 and PARP increased DNA damage and apoptosis. Additionally, the combination of the LIG1 inhibitor L82-G17 with olaparib exhibited synergistic effects. To the best of our knowledge, we validated this combination therapy in vivo for the first time, suppressing tumor growth in a DU145 xenograft model while minimizing toxicity in normal tissues. Immunohistochemical analysis revealed that LIG1 was overexpressed in CRPC tissues, suggesting its potential as a therapeutic target. This study established LIG1 as a novel synthetic lethality-inducing factor in prostate cancer, showing that L82-G17 enhances the efficacy of olaparib, regardless of the BRCA mutation status. These findings suggest that the combination of PARP and LIG1 inhibitors could be a novel therapeutic strategy for mCRPC.

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder caused by pathogenic variants in the TSC1 and TSC2 genes. This condition manifests as diverse symptoms across multiple organ systems and is frequently associated with neuropsychiatric symptoms. However, the relationship between the severity of these manifestations remains poorly understood. This study aimed to elucidate correlations between the severity of various skin and neuropsychiatric symptoms and the underlying TSC1/TSC2 gene pathology.

Integration of human papillomavirus (HPV) into the host genome drives HPV-positive head and neck squamous cell carcinoma (HPV+ HNSCC). Whole-genome sequencing of 51 tumors revealed intratumor heterogeneity of HPV integration, with 44% of breakpoints subclonal, and a biased distribution of integration breakpoints across the HPV genome. Four HPV physical states were identified, with at least 49% of tumors progressing without integration. HPV integration was associated with APOBEC-induced broad genomic instability and focal genomic instability, including structural variants at integration sites. HPV+ HNSCCs exhibited almost no smoking-induced mutational signatures. Heterozygous loss of ataxia-telangiectasia mutated (ATM) was observed in 67% of tumors, with its downregulation confirmed by single-cell RNA sequencing and immunohistochemistry, suggesting ATM haploinsufficiency contributes to carcinogenesis. PI3K activation was the major oncogenic mutation, with JAK-STAT activation in tumors with clonal integration and NF-kappa B activation in those without. These findings provide valuable insights into HPV integration in HPV+ HNSCC.

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology III (PD35)1 May 2024PD35-03 GENOME-WIDE CRISPR SCREENS IDENTIFIED THAT DCLRE1C REGULATES THE RADIOSENSITIVITY OF PROSTATE CANCER Toshiki Oka, Koji Hatano, Keisuke Nimura, Yu Ishizuya, Yuki Horibe, Akihiro Yoshimura, Masaru Tani, Yutong Liu, Sassi Nesrine, Yohei Okuda, Akinaru Yamamoto, Toshihiro Uemura, Gaku Yamamichi, Yoshiyuki Yamamoto, Taigo Kato, Atsunari Kawashima, Kazutoshi Fujita, and Norio Nonomura Toshiki OkaToshiki Oka , Koji HatanoKoji Hatano , Keisuke NimuraKeisuke Nimura , Yu IshizuyaYu Ishizuya , Yuki HoribeYuki Horibe , Akihiro YoshimuraAkihiro Yoshimura , Masaru TaniMasaru Tani , Yutong LiuYutong Liu , Sassi NesrineSassi Nesrine , Yohei OkudaYohei Okuda , Akinaru YamamotoAkinaru Yamamoto , Toshihiro UemuraToshihiro Uemura , Gaku YamamichiGaku Yamamichi , Yoshiyuki YamamotoYoshiyuki Yamamoto , Taigo KatoTaigo Kato , Atsunari KawashimaAtsunari Kawashima , Kazutoshi FujitaKazutoshi Fujita , and Norio NonomuraNorio Nonomura View All Author Informationhttps://doi.org/10.1097/01.JU.0001009396.21455.74.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Radiotherapy is widely used for prostate cancer (PCa), but the therapeutic efficacy varies widely among patients. For more effective radiotherapy, it is necessary to elucidate the molecular mechanisms that regulate radiosensitivity. Androgen receptor (AR) is associated with radiosensitivity in PCa, yet the details remain unclear. Here, we used genome-wide CRISPR screens to identify a gene that regulates radiosensitivity in PCa. We further elucidated how the gene regulates radiosensitivity, including its association with AR. METHODS: PCa cell lines, DU145, 22Rv1, and LNCaP, were used to establish CRISPR/Cas9 cell libraries. The CRISPR library contained 90,709 sgRNAs targeting 18,010 genes. The cell libraries were irradiated (4 or 6 Gy) or untreated (control), and cultured for 11 days. The number of each sgRNA was evaluated by sequencing and compared to controls. Enrichment of sgRNA means that the knockout of its target gene enhances radioresistance, and drop-off of sgRNA means that the knockout of its target gene enhances radiosensitivity. RESULTS: CRISPR screens showed that many cell cycle and apoptosis-related genes were enriched, while many DNA repair pathway genes were dropped off. The most potent radiosensitizing target gene, DCLRE1C, was identified by CRISPR screens of three PCa cell lines. Knockout of DCLRE1C significantly increased γH2AX positive cell rate 2.1-fold and comet tail intensity 1.5-fold after irradiation (IR) in DU145 cells. γH2AX and comet tail reflect DNA double-strand breaks and DNA damage, respectively. Knockout of DCLRE1C reduced the radiation dose by 55% required to achieve 90% cell death, but overexpression of DCLRE1C canceled this phenotype. Similar results were confirmed in 22RV1 and PC3 cells. DCLRE1C was activated following IR by Ser-516 phosphorylation. Notably, DCLRE1C phosphorylation was attenuated by an AR signaling inhibitor. CONCLUSIONS: The most potent radiosensitizing target gene, DCLRE1C, was identified by CRISPR screens. The phosphorylation of DCLRE1C plays a pivotal role in AR-mediated radiosensitivity in PCa. Source of Funding: This research was supported by KAKENHI (grant number: 20K18090, 22K09447) © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e725 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Toshiki Oka More articles by this author Koji Hatano More articles by this author Keisuke Nimura More articles by this author Yu Ishizuya More articles by this author Yuki Horibe More articles by this author Akihiro Yoshimura More articles by this author Masaru Tani More articles by this author Yutong Liu More articles by this author Sassi Nesrine More articles by this author Yohei Okuda More articles by this author Akinaru Yamamoto More articles by this author Toshihiro Uemura More articles by this author Gaku Yamamichi More articles by this author Yoshiyuki Yamamoto More articles by this author Taigo Kato More articles by this author Atsunari Kawashima More articles by this author Kazutoshi Fujita More articles by this author Norio Nonomura More articles by this author Expand All Advertisement PDF downloadLoading ...

mRNA is a promising modality for expressing a protein in vivo. . Drug delivery systems are required for the efficient fi cient transfection of mRNA into cells. In this study, we evaluated several drug delivery systems for transfecting mRNA into tumors. A lipid nanoparticle delivered mRNA to the draining lymph nodes and liver, even by intratumoral injection. A liposome-based system did not consistently provide mRNA for different types of tumor cells. We found that PBS introduced mRNA into several tumors, and calcium ions enhanced the efficiency, fi ciency, particularly in male mice. The circular dichroism spectrometer suggested a structural change in mRNA in PBS. Transmission electron microscopy revealed that calcium ions promoted the formation of mRNA nanoparticles in PBS. Transfection of mRNAs coding OX40-ligand, interleukin (IL)-36g, g , and IL-23 by PBS + calcium ions attenuated tumor growth. Our results indicate that combining PBS with calcium ions promotes the transfection of mRNA into tumors. These data provide information for the development of methods for transfection of mRNA for cancer therapy.

The tumor-infiltrating lymphocyte (TIL) is a crucial factor in controlling tumor growth. A therapeutic method activating TIL is desired for treating patients with metastatic tumors. Here, we show that treating a local tumor with a combination therapy of UV-irradiated hemagglutinating virus of Japan envelope (HVJ-E) plus agonist antibodies, including OX40, against T cell costimulatory molecules induces systemic antitumor effects in a T cell-dependent manner in multiple cancer cell lines. Transcriptome and T cell receptor repertoire analyses revealed that HVJ-E + anti-OX40 antibody treatment activates CD4 and CD8 T cells and promotes T cell trafficking between tumors. These systemic anti-tumor effects required an association between Nkg2d and Nkg2d ligands. Our fi ndings provide insights into how systemic anti-tumor effects are induced and may help the development of therapeutic strategies for eliciting such effects.

Neurofibromatosis 1 is a prevalent hereditary neurocutaneous disorder. Among the clinical phenotypes of neurofibromatosis 1, cutaneous neurofibroma (cNF) and plexiform neurofibroma (pNF) have distinct clinical manifestations, and pNF should be closely monitored owing to its malignant potential. However, the detailed distinct features of neurofibromatosis 1 phenotypes remain unknown. To determine whether the transcriptional features and microenvironment of cNF and pNF differ, single-cell RNA sequencing was performed on isolated cNF and pNF cells from the same patient. Six cNF and five pNF specimens from different subjects were also immunohistochemically analyzed. Our findings revealed that cNF and pNF had distinct transcriptional profiles even within the same subject. pNF is enriched in Schwann cells with characteristics similar to those of their malignant counterpart, fibroblasts, with a cancer-associated fibroblast-like phenotype, angiogenic endothelial cells, and M2-like macrophages, whereas cNF is enriched in CD8 T cells with tissue residency markers. The results of immunohistochemical analyses performed on different subjects agreed with those of single-cell RNA sequencing. This study found that cNF and pNF, the different neurofibromatosis phenotypes in neurofibromatosis 1, from the same subject are transcriptionally distinct in terms of the cell types involved, including T cells.Journal of Investigative Dermatology (2024) 144, 133-141; doi:10.1016/j.jid.2023.03.1688

To confirm the hypothesis that the terminal epoxide structure of spliceostatin A may be active through hydrogen bond formation, we designed and synthesized novel ether derivatives of spliceostatin A, whose terminal epoxide is replaced with an ether-based moiety capable of hydrogen bond formation. The results of biological evaluation against AR-V7 and the conformational analysis of each derivative suggest that the terminal epoxide moiety of spliceostatin A expresses its activity through covalent bond formation rather than hydrogen bond formation.

A list of primers used for PCR and gRNA

A list of antibodies used in the study