Therapeutic cancer vaccination against mutant calreticulin in myeloproliferative neoplasms induces expansion of specific T cells in the periphery but specific T cells fail to enrich in the bone marrow

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  • Morten Orebo Holmström
  • Morten Andersen
  • Sofie Traynor
  • Shamaila Munir Ahmad
  • Thomas Landkildehus Lisle
  • Jacob Handlos Grauslund
  • Vibe Skov
  • Lasse Kjær
  • Johnny T. Ottesen
  • Morten Frier Gjerstorff
  • Hasselbalch, Hans K
  • Andersen, Mads Hald

Background: Therapeutic cancer vaccination against mutant calreticulin (CALR) in patients with CALR-mutant (CALRmut) myeloproliferative neoplasms (MPN) induces strong T-cell responses against mutant CALR yet fails to demonstrate clinical activity. Infiltration of tumor specific T cells into the tumor microenvironment is needed to attain a clinical response to therapeutic cancer vaccination. Aim: Determine if CALRmut specific T cells isolated from vaccinated patients enrich in the bone marrow upon completion of vaccination and explore possible explanations for the lack of enrichment. Methods: CALRmut specific T cells from four of ten vaccinated patients were expanded, enriched, and analyzed by T-cell receptor sequencing (TCRSeq). The TCRs identified were used as fingerprints of CALRmut specific T cells. Bone marrow aspirations from the four patients were acquired at baseline and at the end of trial. T cells were enriched from the bone marrow aspirations and analyzed by TCRSeq to identify the presence and fraction of CALRmut specific T cells at the two different time points. In silico calculations were performed to calculate the ratio between transformed cells and effector cells in patients with CALRmut MPN. Results: The fraction of CALRmut specific T cells in the bone marrow did not increase upon completion of the vaccination trial. In general, the T cell repertoire in the bone marrow remains relatively constant through the vaccination trial. The enriched and expanded CALRmut specific T cells recognize peripheral blood autologous CALRmut cells. In silico analyses demonstrate a high imbalance in the fraction of CALRmut cells and CALRmut specific effector T-cells in peripheral blood. Conclusion: CALRmut specific T cells do not enrich in the bone marrow after therapeutic cancer peptide vaccination against mutant CALR. The specific T cells recognize autologous peripheral blood derived CALRmut cells. In silico analyses demonstrate a high imbalance between the number of transformed cells and CALRmut specific effector T-cells in the periphery. We suggest that the high burden of transformed cells in the periphery compared to the number of effector cells could impact the ability of specific T cells to enrich in the bone marrow.

OriginalsprogEngelsk
Artikelnummer1240678
TidsskriftFrontiers in Immunology
Vol/bind14
Antal sider14
ISSN1664-3224
DOI
StatusUdgivet - 2023

Bibliografisk note

Funding Information:
The authors declare that this study received funding from the BRIDGE – Translational Excellence Programme (bridge.ku.dk) at the Faculty of Health and Medical Sciences, University of Copenhagen, funded by the Novo Nordisk Foundation. Grant agreement no. NNF20SA0064340. Sundhedsstyrelsen – sagsnummer 05-0400-18. Sundhedsstyrelsen – sagsnummer 05-0400-50, Det Frie Forskningsråd – sagsnummer 0134-00072B, Kræftens Bekæmpelse– sagsnummer R149-A10159. The funders were not involved in the study design, collection, analysis, interpretation of data, the writing of this article nor the decision to submit it for publication. Acknowledgments

Funding Information:
The authors declare that this study received funding from the BRIDGE – Translational Excellence Programme (bridge.ku.dk) at the Faculty of Health and Medical Sciences, University of Copenhagen, funded by the Novo Nordisk Foundation. Grant agreement no. NNF20SA0064340. Sundhedsstyrelsen – sagsnummer 05-0400-18. Sundhedsstyrelsen – sagsnummer 05-0400-50, Det Frie Forskningsråd – sagsnummer 0134-00072B, Kræftens Bekæmpelse– sagsnummer R149-A10159. The funders were not involved in the study design, collection, analysis, interpretation of data, the writing of this article nor the decision to submit it for publication.

Publisher Copyright:
Copyright © 2023 Holmström, Andersen, Traynor, Ahmad, Lisle, Handlos Grauslund, Skov, Kjær, Ottesen, Gjerstorff, Hasselbalch and Andersen.

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