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Updated: Feb 21, 2026

Paramyxoviruses for Tumor-targeted Immunomodulation: Design and Evaluation Ex Vivo
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Construct Optimization Enables Oncolytic Virus-Mediated Functional Membrane Localization of Calreticulin and

Xinyuan Zhang1,2, Shengfeng Xiong1,2, Song Zhang1,2

  • 1Department of Obstetrics and Gynecology, National Clinical Research Center for Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Human Gene Therapy
|February 19, 2026
PubMed
Summary

Researchers engineered an oncolytic adenovirus (oAd) to express calreticulin (CALR), enhancing tumor cell lysis and stimulating antitumor immune responses. This optimized viral therapy shows promise for improving cancer immunotherapy by modulating the tumor microenvironment.

Keywords:
calreticulinmacrophage polarizationoncolytic adenovirus

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Area of Science:

  • Oncolytic virotherapy
  • Cancer immunotherapy
  • Immunomodulation

Background:

  • Oncolytic viruses are engineered to selectively infect and kill cancer cells.
  • Enhancing viral platforms for improved tumor targeting and immune stimulation is crucial.
  • Calreticulin (CALR) exposure on tumor cells can promote immune recognition and anti-tumor responses.

Purpose of the Study:

  • To optimize an oncolytic adenovirus (oAd) for enhanced tumor cell lysis and immunomodulation.
  • To identify the optimal insertion site for calreticulin (CALR) expression in an oAd platform.
  • To investigate the mechanism of oAd-mediated CALR expression and its functional consequences on immune cells and tumor growth.

Main Methods:

  • Systematic exploration of CALR insertion sites within an oncolytic adenovirus.
  • Genetic engineering of the oAd including E1A CR2 deletion and E3-gp19k replacement with a CMV promoter.
  • Bulk-RNA sequencing (RNA-Seq) to analyze gene expression changes post-transfection.
  • Ex vivo coculture assays with macrophages and patient-derived ovarian cancer spheroids.
  • In vivo studies using Hepa1-6 xenograft models in mice.

Main Results:

  • An optimal oAd variant (oAd-CMV-CALR) was identified, effectively expressing CALR on tumor cell membranes.
  • oAd-CMV-CALR induced endoplasmic reticulum stress and selective replication in retinoblastoma-deficient tumor cells.
  • Transfected macrophages showed enhanced phagocytic capacity and M1-like repolarization.
  • In vivo studies demonstrated suppressed tumor growth, increased CD8+ T-cell infiltration, and favorable safety.
  • The approach showed translational potential in patient-derived ovarian cancer spheroids.

Conclusions:

  • oAd-CMV-CALR represents a promising therapeutic strategy for cancer immunotherapy.
  • This engineered virus effectively combines direct tumor cell killing with potent immune system activation.
  • The findings highlight the potential of oAd-CMV-CALR to modulate the tumor microenvironment and improve treatment outcomes.