Local Application of Autologous Platelet-Rich Fibrin Patch (PRF-P) Suppresses Regulatory T Cell Recruitment in a Murine Glioma Model
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The immunosuppressive microenvironment is one of the major factors promoting the growth of glioblastoma multiforme (GBM). Infiltration of CD4+CD25+Foxp3+ regulatory T cells (Tregs) into the tumor microenvironment plays a significant role in the suppression of the anti-tumor immunity and portends a dismal prognosis for patients. Glioma-mediated secretion of chemo-attractant C-C motif ligand 2 and 22 (CCL2/22) has previously been shown by our group to promote Treg migration in vitro. In this study, we show that a local implantation of platelet-rich fibrin patch (PRF-P) into the brain of GL261 glioma-bearing mice prolonged the survival of affected animals by 42.85% (p = 0.0011). Analysis performed on brain tumor tissue harvested from PRF-P-treated mice revealed a specific decrease in intra-tumoral lymphocytes with a preferential depletion of immunosuppressive Tregs. Importantly, co-culture of GL261 or chemo-attractants (CCL2/22) with PRF-P abrogated Treg migration. Pharmacological blockade of the CCL2/22 interaction with their receptors potentiated the inhibitory effect of PRF-P on Tregs recruitment in culture. Moreover, our findings revealed the soluble CD40 ligand (sCD40L) as a major Treg inhibitory player produced by activated platelets entrapped within the fibrin matrix of the PRF-P. Blockade of sCD40L restored the migratory capacity of Tregs, emphasizing the role of PRF-P in preventing the Treg migration to glioma tissue. Our findings highlight autologous PRF-P as a personalized, Treg-selective suppression platform that can potentially supplement and enhance the efficacy of glioma therapies.
KeywordsGlioblastoma multiforme Immunosuppressive microenvironment Platelet-rich fibrin patch Regulatory T cells and glioma immune therapies
The authors would like to thank Irina Balyasnikova and Atique U. Ahmed for their comments and suggestions.
W.K.P., K.C.P., and M.S.L. conceived the study. W.K.P., K.C.P., J.M., J.W.K., A.R., D.K., D.Y., C.L.C., J.R.K., and A.C designed, performed, and analyzed the experiments. A.L.R. performed all animal breeding for the study. Y.H. and P.Z provided assistance with animal surgeries and reagent preparations. M.S.L. provided critical feedback, contributed to manuscript preparation, and oversaw the research program. All authors listed reviewed the manuscript and provided feedback with writing and revisions.
This work is supported by National Cancer Institute Outstanding Investigator Award from NIH/National Cancer Institute to M.S.L. (R35CA197725) and by a grant from NIH/National Cancer Institute to M.S.L. (R01 NS087990).
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