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Raza Ali Naqvi, Imran Ahmad, Araceli Valverde, Afsar R Naqvi, Long noncoding RNAs RN7SK and GAS5 Regulate Macrophage Innate Immune Function and Polarization, The Journal of Immunology, Volume 206, Issue 1_Supplement, May 2021, Page 64.10, https://doi.org/10.4049/jimmunol.206.Supp.64.10
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Abstract
Macrophages (Mϕ) are polarized in two phenotypes: classically activated M1 phenotype and alternatively activated M2 phenotype to marginalize the pathogenic/non-pathogenic threat. Burgeoning reports are now elucidating the regulatory role of lncRNAs in various immune process. In this study, we aimed to identify the lncRNAs that mediate Mϕ function and polarization.
CD14+ monocytes isolated from PBMCs and differentiated to M1 and M2Mϕ, using GM-CSF or M-CSF, respectively. Expression of 88 different lncRNAs was examined by RT-qPCR. Cells transfected with siRNAs targeting RN7SK, GAS5, IPW and ZFAS1 were examined for Mϕ polarization markers and innate immune functions (phagocytosis and antigen uptake/processing).
Our results show differential expression of seventeen lncRNAs in M2Mϕ compared to monocytes including RN7SK, GAS5, IPW and ZFAS1. Knockdown of RN7SK and GAS5 showed downregulation of M2 surface markers (CD163, CD206 or Dectin) and concomitant increase in M1 markers (MHC II or CD23). M1/M2 markers were not impacted by IPW or ZFAS1 knockdown. mRNA levels of polarization markers were not effected by RN7SK or GAS5 RNAi. We observed enhanced antigen uptake and processing both in M1/M2 Mϕ transfected with siRNA targeting GAS5 and RN7SK but not IPW and ZFAS1. In addition, knockdown of RN7SK significantly augmented labelled E.coli uptake by M1/M2 Mϕ, while no significant difference was noticed in GAS5 silenced cells.
This study clearly deciphered that remarkable changes in lncRNA occur during monocyte to Mϕ differentiation, polarization and regulate integral innate functions of Mϕ. Manipulation of lncRNAs could be tested in therapeutic reversal of pro and anti-inflammatory pathologies.