Long-term intravital imaging of the multicolor-coded tumor microenvironment during combination immunotherapy

  1. Shuhong Qi
  2. Hui Li
  3. Lisen Lu
  4. Zhongyang Qi
  5. Lei Liu
  6. Lu Chen
  7. Guanxin Shen
  8. Ling Fu
  9. Qingming Luo  Is a corresponding author
  10. Zhihong Zhang  Is a corresponding author
  1. Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, China
  2. Tongji Medical College, China

Abstract

The combined-immunotherapy of adoptive cell therapy (ACT) and cyclophosphamide (CTX) is one of the most efficient treatments for melanoma patients. However, synergistic effects of CTX and ACT on the spatio-temporal dynamics of immunocytes in vivo have not been described. Here, we visualized key cell events of immunotherapy-elicited immunoreactions in a multicolor-coded tumor microenvironment, and then established an optimal strategy of metronomic combined-immunotherapy to enhance anti-tumor efficacy. Intravital imaging data indicated that regulatory T cells formed an 'immunosuppressive ring' around a solid tumor. The CTX-ACT combined-treatment elicited synergistic immunoreactions in tumor areas, which included relieving the immune suppression, triggering the transient activation of endogenous tumor-infiltrating immunocytes, increasing the accumulation of adoptive cytotoxic T lymphocytes, and accelerating the infiltration of dendritic cells. These insights into the spatio-temporal dynamics of immunocytes are beneficial for optimizing immunotherapy and provide new approaches for elucidating the mechanisms underlying the involvement of immunocytes in cancer immunotherapy.

Article and author information

Author details

  1. Shuhong Qi

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Hui Li

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Lisen Lu

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Zhongyang Qi

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Lei Liu

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Lu Chen

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Guanxin Shen

    Department of Immunology, Tongji Medical College, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Ling Fu

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Qingming Luo

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    For correspondence
    qluo@mail.hust.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
  10. Zhihong Zhang

    Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China
    For correspondence
    czyzzh@mail.hust.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5227-8926

Funding

National Natural Science Foundation of China (91442201)

  • Shuhong Qi
  • Lisen Lu
  • Lei Liu
  • Zhihong Zhang

Ministry of Science and Technology of the People's Republic of China (2011CB910401)

  • Shuhong Qi
  • Hui Li
  • Lei Liu
  • Qingming Luo
  • Zhihong Zhang

National Natural Science Foundation of China (61421064)

  • Ling Fu
  • Qingming Luo
  • Zhihong Zhang

Ministry of Education of the People's Republic of China (2015ZDTD014)

  • Shuhong Qi
  • Lisen Lu
  • Lei Liu
  • Zhihong Zhang

Ministry of Science and Technology of the People's Republic of China (Director fund of Wuhan National Laboratory for Optoelectronics)

  • Shuhong Qi
  • Lisen Lu
  • Lei Liu
  • Qingming Luo
  • Zhihong Zhang

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of Hubei Provincial Animal Care and Use Committee. The protocol was approved by the Animal Experimentation Ethics Committee of Huazhong University of Science and Technology (reference number: 452). All surgery was performed under ketamine and xylazine, and all intravital imaging experiments were performed under 1-3 % isoflurane in oxygen, every effort was made to minimize suffering.

Reviewing Editor

  1. Xuetao Cao, Zhejiang University School of Medicine, China

Publication history

  1. Received: January 28, 2016
  2. Accepted: October 17, 2016
  3. Accepted Manuscript published: November 18, 2016 (version 1)
  4. Version of Record published: December 20, 2016 (version 2)

Copyright

© 2016, Qi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,237
    Page views
  • 734
    Downloads
  • 33
    Citations

Article citation count generated by polling the highest count across the following sources: Scopus, Crossref, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Shuhong Qi
  2. Hui Li
  3. Lisen Lu
  4. Zhongyang Qi
  5. Lei Liu
  6. Lu Chen
  7. Guanxin Shen
  8. Ling Fu
  9. Qingming Luo
  10. Zhihong Zhang
(2016)
Long-term intravital imaging of the multicolor-coded tumor microenvironment during combination immunotherapy
eLife 5:e14756.
https://doi.org/10.7554/eLife.14756

Further reading

    1. Cancer Biology
    Ning Yang, Xuebo Lu ... Kangdong Liu
    Research Article Updated

    Human esophageal cancer has a global impact on human health due to its high incidence and mortality. Therefore, there is an urgent need to develop new drugs to treat or prevent the prominent pathological subtype of esophageal cancer, esophageal squamous cell carcinoma (ESCC). Based upon the screening of drugs approved by the Food and Drug Administration, we discovered that Arbidol could effectively inhibit the proliferation of human ESCC in vitro. Next, we conducted a series of cell-based assays and found that Arbidol treatment inhibited the proliferation and colony formation ability of ESCC cells and promoted G1-phase cell cycle arrest. Phosphoproteomics experiments, in vitro kinase assays and pull-down assays were subsequently performed in order to identify the underlying growth inhibitory mechanism. We verified that Arbidol is a potential ataxia telangiectasia and Rad3-related (ATR) inhibitor via binding to ATR kinase to reduce the phosphorylation and activation of minichromosome maintenance protein 2 at Ser108. Finally, we demonstrated Arbidol had the inhibitory effect of ESCC in vivo by a patient-derived xenograft model. All together, Arbidol inhibits the proliferation of ESCC in vitro and in vivo through the DNA replication pathway and is associated with the cell cycle.

    1. Cancer Biology
    2. Computational and Systems Biology
    Pan Cheng, Xin Zhao ... Teresa Davoli
    Research Article

    How cells control gene expression is a fundamental question. The relative contribution of protein-level and RNA-level regulation to this process remains unclear. Here, we perform a proteogenomic analysis of tumors and untransformed cells containing somatic copy number alterations (SCNAs). By revealing how cells regulate RNA and protein abundances of genes with SCNAs, we provide insights into the rules of gene regulation. Protein complex genes have a strong protein-level regulation while non-complex genes have a strong RNA-level regulation. Notable exceptions are plasma membrane protein complex genes, which show a weak protein-level regulation and a stronger RNA-level regulation. Strikingly, we find a strong negative association between the degree of RNA-level and protein-level regulation across genes and cellular pathways. Moreover, genes participating in the same pathway show a similar degree of RNA- and protein-level regulation. Pathways including translation, splicing, RNA processing, and mitochondrial function show a stronger protein-level regulation while cell adhesion and migration pathways show a stronger RNA-level regulation. These results suggest that the evolution of gene regulation is shaped by functional constraints and that many cellular pathways tend to evolve one predominant mechanism of gene regulation at the protein level or at the RNA level.