Molecular targets for therapy

Anti-CD69 therapy induces rapid mobilization and high proliferation of HSPCs through S1P and mTOR

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CD69 regulates lymphocyte egress from the thymus and lymph nodes through cis-interactions and the downregulation of surface sphingosine-1-phosphate (S1P) receptor-1 (S1P1). However, its role in the regulation of cell egress from bone marrow has not been extensively studied. We show here that CD69 targeting induced rapid and massive mobilization of BM leukocytes, which was inhibited by desensitization to S1P with FTY720. This mobilization was reproduced with anti-human CD69 mAb treatment of mice expressing human CD69. In this strain, the mobilization occurred to the same extent as that induced by AMD3100. The anti-human CD69 treatment highly increased LSK and CLP cell proliferation and numbers, both in the periphery and in the BM, and also augmented S1P1 and CXCR4 expression. Additionally, increased mTOR, p70S6K, S6, and 4E-BP1 phosphorylation was detected after in vivo anti-CD69 treatment in the bone marrow. Importantly, mTOR inhibition with rapamycin inhibited anti-huCD69-induced mobilization of hematopoietic stem and progenitor cells (HSPCs). Together, our results indicated that CD69 targeting induces not only mobilization but also high proliferation of HSPCs, and thus is crucial for precursor cell replenishment over time. These results suggest that anti-CD69 mAbs are putative novel candidates for mobilization strategies.

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The study was supported by the Instituto de Salud Carlos III MPY 1366/13 and MPY 1346/16. ML was supported by SAF2015-74112-JIN from MINECO. GS was supported by ERC 260464, EFSD 2030, MINECO-FEDER SAF2016-79126-R, and Comunidad de Madrid S2010/BMD-2326. The CNIC is supported by the Ministerio de Economía y Competitividad and the Pro-CNIC Foundation. The Pro-CNIC Foundation is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505). We are grateful to Dra Ana Justel, of UAM, for help in statistical analysis. We thank Daniel Baizan, Cristina Pintos, and Maria Clemente for mouse husbandry.

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Correspondence to Pilar Lauzurica.

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