TY - JOUR
T1 - Significant role for microRNA-21 affecting toll-like receptor pathway in primary graft dysfunction after human lung transplantation
AU - Xu, Zhongping
AU - Sharma, Monal
AU - Gelman, Andrew
AU - Hachem, Ramsey
AU - Mohanakumar, Thalachallour
N1 - Publisher Copyright:
© 2017 International Society for Heart and Lung Transplantation
PY - 2017/3/1
Y1 - 2017/3/1
N2 - Background MicroRNAs (miRNAs) were recently identified as modulators of immune responses after human lung transplantation (LTx). This study was undertaken to assess the contribution of miRNAs to the pathogenesis of primary graft dysfunction (PGD) after LTx. Methods Of the 39 recipients, 14 (35.9%) developed Grade 3 PGD (i.e., severe PGD) within the first 72 hours of LTx. The remaining 25 recipients (64.1%) had Grade 2 or less PGD, and served as the control group. miRNAs were isolated from cells purified by bronchoalveolar lavage (BAL). Bioinformatic prediction and validation by luciferase reporter assays were performed to identify targets regulated by miR-21. Transfection of human monocytic cell line (THP-1) was conducted to determine miR-21’s cellular function. Results Pilot miRNA profiling of donor BAL samples before implantation in PGD (n = 6) revealed significant upregulation in 44 miRNAs and downregulation in 80 miRNAs compared with control (n = 6). Validation using a separate cohort demonstrated significant underexpression of miR-21 in patients with severe PGD. Furthermore, underexpression of miR-21 levels was negatively correlated with clinical PGD grades (Grade 2 PGD vs Grade 0 PGD: p = 0.042; Grade 3 PGD vs Grade 0 PGD: p = 0.004). Molecular analysis demonstrated that miR-21 targeted key components in the toll-like receptor (TLR) signaling pathway, including TLR4, IRAK3 and CXCL10. Further, incubation of THP-1 cells with cell-free BAL from severe PGD resulted in transactivation of inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). In contrast, increased expression of miR-21 resulted in marked suppression of IL-1-β and TNF-α production. Conclusions Underexpression of miR-21 may lead to the development of severe PGD by activating key components of the TLR pathway.
AB - Background MicroRNAs (miRNAs) were recently identified as modulators of immune responses after human lung transplantation (LTx). This study was undertaken to assess the contribution of miRNAs to the pathogenesis of primary graft dysfunction (PGD) after LTx. Methods Of the 39 recipients, 14 (35.9%) developed Grade 3 PGD (i.e., severe PGD) within the first 72 hours of LTx. The remaining 25 recipients (64.1%) had Grade 2 or less PGD, and served as the control group. miRNAs were isolated from cells purified by bronchoalveolar lavage (BAL). Bioinformatic prediction and validation by luciferase reporter assays were performed to identify targets regulated by miR-21. Transfection of human monocytic cell line (THP-1) was conducted to determine miR-21’s cellular function. Results Pilot miRNA profiling of donor BAL samples before implantation in PGD (n = 6) revealed significant upregulation in 44 miRNAs and downregulation in 80 miRNAs compared with control (n = 6). Validation using a separate cohort demonstrated significant underexpression of miR-21 in patients with severe PGD. Furthermore, underexpression of miR-21 levels was negatively correlated with clinical PGD grades (Grade 2 PGD vs Grade 0 PGD: p = 0.042; Grade 3 PGD vs Grade 0 PGD: p = 0.004). Molecular analysis demonstrated that miR-21 targeted key components in the toll-like receptor (TLR) signaling pathway, including TLR4, IRAK3 and CXCL10. Further, incubation of THP-1 cells with cell-free BAL from severe PGD resulted in transactivation of inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). In contrast, increased expression of miR-21 resulted in marked suppression of IL-1-β and TNF-α production. Conclusions Underexpression of miR-21 may lead to the development of severe PGD by activating key components of the TLR pathway.
KW - lung transplantation
KW - microRNAs
KW - primary graft dysfunction
UR - http://www.scopus.com/inward/record.url?scp=85008930057&partnerID=8YFLogxK
U2 - 10.1016/j.healun.2016.08.028
DO - 10.1016/j.healun.2016.08.028
M3 - Article
C2 - 27773452
AN - SCOPUS:85008930057
SN - 1053-2498
VL - 36
SP - 331
EP - 339
JO - Journal of Heart and Lung Transplantation
JF - Journal of Heart and Lung Transplantation
IS - 3
ER -