Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results

IntroductionIntratumoral immunotherapy delivers immune-modifying therapeutic agents directly into the tumor microenvironment (TME), stimulating both local and systemic immune responses. In this pilot study, we evaluated the immunomodulatory effects of cowpea mosaic virus (CPMV) particles, which prim...

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Main Authors: Akansha Singh, Jessica Fernanda Affonso de Oliveira, Jessica Schrader, Deepan Kishore, Sri Vidhya Chandrasekar, Steven Fiering, Nicole F. Steinmetz, Ashish Ranjan
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1566394/full
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author Akansha Singh
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Schrader
Deepan Kishore
Sri Vidhya Chandrasekar
Steven Fiering
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Ashish Ranjan
author_facet Akansha Singh
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Schrader
Deepan Kishore
Sri Vidhya Chandrasekar
Steven Fiering
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Ashish Ranjan
author_sort Akansha Singh
collection DOAJ
description IntroductionIntratumoral immunotherapy delivers immune-modifying therapeutic agents directly into the tumor microenvironment (TME), stimulating both local and systemic immune responses. In this pilot study, we evaluated the immunomodulatory effects of cowpea mosaic virus (CPMV) particles, which primarily activates innate immunity, and calreticulin nanoparticle (CRT-NP), which enhance immunostimulatory signals of immunogenic cell death in canine cancers. The study focused on their potential to induce local and systemic antitumor immune responses and achieve tumor control.MethodCPMV was obtained through the mechanical inoculation of Vigna unguiculata, while CRT-NP was generated using cationic liposomes loaded with a CRT-expressing plasmid. Nine canine patients with oral melanoma, soft-tissue sarcoma (STS), and mammary gland carcinoma received CPMV or CRT-NP via intratumoral route. CPMV and CRT-NP was administered weekly at three-five intratumoral locations. To evaluate antitumor immune responses, biopsies and blood samples were obtained prior to treatment and during follow-up visits, extending up to one week after the final treatment. Endpoints included serum cytokine analysis, tumor transcriptomics via NanoString, immune cell profiling in blood and tumor biopsies, and efficacy assessment using RECIST criteria.ResultCPMV exhibited an icosahedral shape (~30 nm), while CRT-NP were spherical (~300 nm). CPMV induced stable disease (SD) in two of three melanoma and STS patients, with partial response (PR) in the third. CRT-NP induced SD in two of three patients, with one STS patient achieving partial remission. Post-treatment NanoString and flow cytometry analyses revealed a shift in the tumor microenvironment toward a more immunostimulatory state, marked by increased neutrophils and CD8+ T cells. CPMV, in particular, upregulated genes involved in antigen processing and immune activation while enhancing IFNγ+ CD4+ and CD8+ T cell populations. CRT-NP reduced Tregs in the TME. Further, serum cytokine levels, such as MCP-1, GM-CSF, IL-2, IL-6, IL-7 and IL-18, correlated with tumor growth independent of various treatments.DiscussionOur findings suggest that CPMV and CRT-NP, which activate distinct immunologic pathways, safely modulate the TME contributing to disease stabilization in spontaneous canine cancers. These results support the need for larger-scale trials to address current limitations, differentiate tumor-agnostic versus treatment-specific effects, and evaluate long-term clinical outcomes, including overall survival (OS).
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spelling doaj-art-ef34b2ab3f96473abc1a7b0eac67950b2025-08-20T02:13:35ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.15663941566394Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early resultsAkansha Singh0Jessica Fernanda Affonso de Oliveira1Jessica Fernanda Affonso de Oliveira2Jessica Fernanda Affonso de Oliveira3Jessica Schrader4Deepan Kishore5Sri Vidhya Chandrasekar6Steven Fiering7Nicole F. Steinmetz8Nicole F. Steinmetz9Nicole F. Steinmetz10Nicole F. Steinmetz11Nicole F. Steinmetz12Nicole F. Steinmetz13Nicole F. Steinmetz14Nicole F. Steinmetz15Ashish Ranjan16Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK, United StatesAiiso Yufeng Li Family Department of Chemical and Nanoengineering, University of California, San Diego, La Jolla, CA, United StatesShu and K.C. Chien and Peter Farrell Collaboratory, University of California, San Diego, La Jolla, CA, United StatesCenter for Nano-ImmunoEngineering, University of California, San Diego, La Jolla, CA, United StatesNeel Veterinary Hospital, Oklahoma City, OK, United StatesNeel Veterinary Hospital, Oklahoma City, OK, United StatesDepartment of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK, United StatesDepartment of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Lebanon, NH, United StatesAiiso Yufeng Li Family Department of Chemical and Nanoengineering, University of California, San Diego, La Jolla, CA, United StatesShu and K.C. Chien and Peter Farrell Collaboratory, University of California, San Diego, La Jolla, CA, United StatesCenter for Nano-ImmunoEngineering, University of California, San Diego, La Jolla, CA, United StatesDepartment of Bioengineering, University of California, San Diego, La Jolla, CA, United StatesDepartment of Radiology, University of California, San Diego, La Jolla, CA, United StatesInstitute for Materials Discovery and Design, University of California, San Diego, La Jolla, CA, United States0Moores Cancer Center, University of California, San Diego, La Jolla, CA, United States1Center for Engineering in Cancer, Institute of Engineering Medicine, University of California, San Diego, La Jolla, CA, United StatesDepartment of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK, United StatesIntroductionIntratumoral immunotherapy delivers immune-modifying therapeutic agents directly into the tumor microenvironment (TME), stimulating both local and systemic immune responses. In this pilot study, we evaluated the immunomodulatory effects of cowpea mosaic virus (CPMV) particles, which primarily activates innate immunity, and calreticulin nanoparticle (CRT-NP), which enhance immunostimulatory signals of immunogenic cell death in canine cancers. The study focused on their potential to induce local and systemic antitumor immune responses and achieve tumor control.MethodCPMV was obtained through the mechanical inoculation of Vigna unguiculata, while CRT-NP was generated using cationic liposomes loaded with a CRT-expressing plasmid. Nine canine patients with oral melanoma, soft-tissue sarcoma (STS), and mammary gland carcinoma received CPMV or CRT-NP via intratumoral route. CPMV and CRT-NP was administered weekly at three-five intratumoral locations. To evaluate antitumor immune responses, biopsies and blood samples were obtained prior to treatment and during follow-up visits, extending up to one week after the final treatment. Endpoints included serum cytokine analysis, tumor transcriptomics via NanoString, immune cell profiling in blood and tumor biopsies, and efficacy assessment using RECIST criteria.ResultCPMV exhibited an icosahedral shape (~30 nm), while CRT-NP were spherical (~300 nm). CPMV induced stable disease (SD) in two of three melanoma and STS patients, with partial response (PR) in the third. CRT-NP induced SD in two of three patients, with one STS patient achieving partial remission. Post-treatment NanoString and flow cytometry analyses revealed a shift in the tumor microenvironment toward a more immunostimulatory state, marked by increased neutrophils and CD8+ T cells. CPMV, in particular, upregulated genes involved in antigen processing and immune activation while enhancing IFNγ+ CD4+ and CD8+ T cell populations. CRT-NP reduced Tregs in the TME. Further, serum cytokine levels, such as MCP-1, GM-CSF, IL-2, IL-6, IL-7 and IL-18, correlated with tumor growth independent of various treatments.DiscussionOur findings suggest that CPMV and CRT-NP, which activate distinct immunologic pathways, safely modulate the TME contributing to disease stabilization in spontaneous canine cancers. These results support the need for larger-scale trials to address current limitations, differentiate tumor-agnostic versus treatment-specific effects, and evaluate long-term clinical outcomes, including overall survival (OS).https://www.frontiersin.org/articles/10.3389/fimmu.2025.1566394/fullcowpea mosaic virus (CPMV) nanoparticlescalreticulin nanoparticleintratumoral immunotherapymelanomasarcomaimmunomodulation
spellingShingle Akansha Singh
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Fernanda Affonso de Oliveira
Jessica Schrader
Deepan Kishore
Sri Vidhya Chandrasekar
Steven Fiering
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Nicole F. Steinmetz
Ashish Ranjan
Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
Frontiers in Immunology
cowpea mosaic virus (CPMV) nanoparticles
calreticulin nanoparticle
intratumoral immunotherapy
melanoma
sarcoma
immunomodulation
title Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
title_full Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
title_fullStr Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
title_full_unstemmed Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
title_short Immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors: early results
title_sort immunomodulatory effects of intratumoral cowpea mosaic virus and calreticulin nanoparticles in canine tumors early results
topic cowpea mosaic virus (CPMV) nanoparticles
calreticulin nanoparticle
intratumoral immunotherapy
melanoma
sarcoma
immunomodulation
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1566394/full
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