Supplementary Materials1: Table S1
May 13, 2021
Supplementary Materials1: Table S1. of action is still unknown. Here, we present in vivo genetic evidence showing that Whsc1 plays an important role at several points of hematopoietic development. Particularly, our results demonstrate that both differentiation and function of plays an important function in hematopoiesis in vivo, demonstrating a role for in the immunodeficiency in Wolf-Hirschhorn Syndrome. gene (and is also involved in other pathologies affecting B lymphocytes, like multiple myeloma (Chesi et al., 1998; Stec et al., 1998) and child years B cell acute lymphoblastic leukemias (Huether et al., 2014; Jaffe et al., 2013). Furthermore, it belongs to the protein family of Nuclear SET [Su(var)3C9, Enhancer-of-zeste, Trithorax] Domain name proteins (NSD) whose other users are also involved in developmental and tumoral pathologies (Morishita and di Luccio, 2011). The WHSC1 protein Tasisulam sodium contains a SET domain name that confers it with histone-methyltransferase activity (Marango et al., 2008; Stec et al., 1998). Its most important in-vivo activity is to mediate H3K36 mono- and di-methylation (Kuo et al., 2011), therefore acting as an epigenetic regulator (Kuo et al., 2011). Methylation at H3K36 has been associated with Tasisulam sodium regulation of transcription, splicing, DNA replication and DNA repair (Wagner and Carpenter, 2012). So far, a specific role for WHSC1 in the immune defects associated to WHS patients has not been proven and, in general, the functions of the users of the NSD family in normal hematopoiesis have not been investigated, even though they are recurrently involved in hematopoietic malignancies (Shilatifard and Hu, 2016). Here, we present in vivo genetic evidence showing that deficiency impairs normal hematopoietic development at several stages and lineages, and particularly affects B cell differentiation and mature B cell function. These findings reveal the role of Whsc1 as a player in hematopoietic development and also show that many of the immune defects associated to WHS can be directly attributed to the reduced levels of gene, we first analyzed the hematopoietic development in heterozygous mice (Nimura et al., 2009). We could not identify any major hematopoietic switch in leads to an impairment in lymphoid development that, under normal conditions, only manifests as the mice get older. Whsc1 is required for normal hematopoietic development Given that is not purely essential for the development of any KRT17 of the hematopoietic Tasisulam sodium lineages. However, there were differences in the reconstitutive capacity of erythroid progenitors (erythroblasts). Within (Physique 1G). Also in the spleen there was a strong increase in the percentages of erythroblasts (Physique S3A and Physique 1G), suggesting the presence of extramedullary erythropoiesis. Finally, these alterations also led to a reduction of total cellularity in the spleen of in erythropoiesis in the long term can already be seen in secondary recipients by hematic counting, which shows reductions in reddish blood cells, hemoglobin, hematocrit and platelets (Physique S3B). All these effects show an impairment in the repopulation capacity of dose-dependent, reduction in the percentages of LSK cells in the bone marrow. Open in a separate window Physique 3 Impaired functionality of is required for an efficient CSR to most of the isotypes, providing a model that really recapitulates one of the most severe complications confronted by WHS patients. Open in a separate window Physique 4 Impaired CSR in led to important malfunctions, we performed in vivo BrdU labellings. The results showed that, in the BM, both B cells at all the different developmental stages (Physique 5B,F) and LSK cells (Physique 5C) also offered an increase in the number of BrdU+ S-phase cells, while cluster (Physique S6A and Furniture S1C2). These developmental genes, although of great importance to the morphogenetic pathways affected in WHS patients, do not explain the B cell phenotypes that we have described. However, by using pathway analysis, we can see Tasisulam sodium that many key processes like cell cycle, splicing, ribosome synthesis, DNA replication or DNA repair are very significantly altered in proliferating (Physique 6C), confirming an impairment in the advancement of the replication fork, coupled with the activation.