Barbosa Christiano M V, Fock Ricardo Ambrósio, Hastreiter Araceli Aparecida, Reutelingsperger Cris, Perretti Mauro, Paredes-Gamero Edgar J, Farsky Sandra H P
1Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, Sao Paulo, Brazil.
2CARIM School for Cardiovascular Diseases, Maastricht University, 6200 MD Maastricht, The Netherlands.
Cell Death Discov. 2019 Sep 23;5:135. doi: 10.1038/s41420-019-0215-1. eCollection 2019.
Annexin A1 (AnxA1) modulates neutrophil life span and bone marrow/blood cell trafficking thorough activation of formyl-peptide receptors (FPRs). Here, we investigated the effect of exogenous AnxA1 on haematopoiesis in the mouse. Treatment of C57BL/6 mice with recombinant AnxA1 (rAnxA1) reduced the granulocyte-macrophage progenitor (GMP) population in the bone marrow, enhanced the number of mature granulocytes Gr-1Mac-1 in the bone marrow as well as peripheral granulocytic neutrophils and increased expression of mitotic cyclin B1 on hematopoietic stem cells (HSCs)/progenitor cells (LinSca-1c-Kit: LSK). These effects were abolished by simultaneous treatment with Boc-2, an FPR pan-antagonist. In in vitro studies, rAnxA1 reduced both HSC (LSKCD90FLK-2) and GMP populations while enhancing mature cells (Gr1Mac1). Moreover, rAnxA1 induced LSK cell proliferation (Ki67), increasing the percentage of cells in the S/G2/M cell cycle phases and reducing Notch-1 expression. Simultaneous treatment with WRW4, a selective FPR2 antagonist, reversed the in vitro effects elicited by rAnxA1. Treatment of LSK cells with rAnxA1 led to phosphorylation of PCLγ2, PKC, RAS, MEK, and ERK1/2 with increased expression of NFAT2. In long-term bone marrow cultures, rAnxA1 did not alter the percentage of LSK cells but enhanced the Gr-1Mac-1 population; treatment with a PLC (U73122), but not with a PKC (GF109203), inhibitor reduced rAnxA1-induced phosphorylation of ERK1/2 and Elk1. Therefore, we identify here rAnxA1 as an inducer of HSC/progenitor cell differentiation, favouring differentiation of the myeloid/granulocytic lineage, via Ca/MAPK signalling transduction pathways.
膜联蛋白A1（AnxA1）通过甲酰肽受体（FPRs）的激活来调节中性粒细胞寿命以及骨髓/血细胞运输。在此，我们研究了外源性AnxA1对小鼠造血作用的影响。用重组AnxA1（rAnxA1）处理C57BL/6小鼠可减少骨髓中粒细胞-巨噬细胞祖细胞（GMP）群体，增加骨髓中成熟粒细胞Gr-1Mac-1以及外周粒细胞性中性粒细胞的数量，并增加造血干细胞（HSCs）/祖细胞（LinSca-1c-Kit：LSK）上有丝分裂细胞周期蛋白B1的表达。同时用FPR泛拮抗剂Boc-2处理可消除这些作用。在体外研究中，rAnxA1减少了HSC（LSKCD90FLK-2）和GMP群体，同时增加了成熟细胞（Gr1Mac1）。此外，rAnxA1诱导LSK细胞增殖（Ki67），增加处于S/G2/M细胞周期阶段的细胞百分比并降低Notch-1表达。同时用选择性FPR2拮抗剂WRW4处理可逆转rAnxA1引发的体外作用。用rAnxA1处理LSK细胞导致PCLγ2、PKC、RAS、MEK和ERK1/2磷酸化，同时NFAT2表达增加。在长期骨髓培养中，rAnxA1未改变LSK细胞百分比，但增加了Gr-1Mac-1群体；用磷脂酶C（U73122）而非蛋白激酶C（GF109203）抑制剂处理可降低rAnxA1诱导的ERK1/2和Elk1磷酸化。因此，我们在此确定rAnxA1是HSC/祖细胞分化的诱导剂，通过钙/丝裂原活化蛋白激酶信号转导途径促进髓系/粒细胞系的分化。
