The epigenetic regulator RINF (CXXC5) maintains SMAD7 expression in human immature erythroid cells and sustains red blood cell expansion

• Published in: Haematologica (Roma) 2020-11, Vol.107 (1), p.268-283
• Main Authors: Astori, Audrey, Matherat, Gabriel, Munoz, Isabelle, Gautier, Emilie-Fleur, Surdez, Didier, Zermati, Yaël, Verdier, Frédérique, Zaidi, Sakina, Feuillet, Vincent, Kadi, Amir, Lauret, Evelyne, Delattre, Olivier, Lefèvre, Carine, Fontenay, Michaela, Ségal-Bendirdjian, Evelyne, Dusanter-Fourt, Isabelle, Bouscary, Didier, Hermine, Olivier, Mayeux, Patrick, Pendino, Frédéric
• Format: Article
• Language: English
• Subjects: Life Sciences
• ISSN: 0390-6078; 1592-8721
• DOI: 10.3324/haematol.2020.263558

The gene CXXC5, encoding a retinoid-inducible nuclear factor (RINF), is located within a region at 5q31.2 commonly deleted in myelodysplastic syndrome and adult acute myeloid leukemia. RINF may act as an epigenetic regulator and has been proposed as a tumor suppressor in hematopoietic malignancies. However, functional studies in normal hematopoiesis are lacking, and its mechanism of action is unknown. Here, we evaluated the consequences of RINF silencing on cytokine-induced erythroid differentiation of human primary CD34+ progenitors. We found that RINF is expressed in immature erythroid cells and that RINF-knockdown accelerated erythropoietin-driven maturation, leading to a significant reduction (~45%) in the number of red blood cells, without affecting cell viability. The phenotype induced by RINF-silencing was dependent on tumor growth factor b (TGFb) and mediated by SMAD7, a TGFb-signaling inhibitor. RINF upregulates SMAD7 expression by direct binding to its promoter and we found a close correlation between RINF and SMAD7 mRNA levels both in CD34+ cells isolated from bone marrow of healthy donors and myelodysplastic syndrome patients with del(5q). Importantly, RINF knockdown attenuated SMAD7 expression in primary cells and ectopic SMAD7 expression was sufficient to prevent the RINF knockdown-dependent erythroid phenotype. Finally, RINF silencing affects 5’-hydroxymethylation of human erythroblasts, in agreement with its recently described role as a TET2-anchoring platform in mouse. Collectively, our data bring insight into how the epigenetic factor RINF, as a transcriptional regulator of SMAD7, may fine-tune cell sensitivity to TGFb superfamily cytokines and thus play an important role in both normal and pathological erythropoiesis.