Page 83 - Haematologica April 2020

Red Cell Biology & its Disorders
ARHGEF12 regulates erythropoiesis and is involved in erythroid regeneration after chemotherapy in acute lymphoblastic leukemia patients
Yangyang Xie,1* Li Gao,2* Chunhui Xu,3 Liming Chu,3,7 Lei Gao,4 Ruichi Wu,1 Yu Liu,1 Ting Liu,1 Xiao-jian Sun,5 Ruibao Ren,5 Jingyan Tang,1 Yi Zheng,6 Yong Zhou7 and Shuhong Shen1
1Key Lab of Pediatrics Hematology/Oncology, Ministry of Health, Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiao Tong University, Shanghai, China; 2Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China; 3Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China; 4CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Beijing, China; 5State Key Laboratory for Medical Genomics, Shanghai Institute of Hematology, Ruijin Hospital, Shanghai, China; 6Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Research Foundation, Cincinnati, OH, USA and 7CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, China
*YX and LG contributed equally to this work.
ABSTRACT
Hematopoiesis is a finely regulated process in vertebrates under both homeostatic and stress conditions. By whole exome sequencing, we studied the genomics of acute lymphoblastic leukemia (ALL) patients who needed multiple red blood cell (RBC) transfusions after inten- sive chemotherapy treatment. ARHGEF12, encoding a RhoA guanine nucleotide exchange factor, was found to be associated with chemothera- py-induced anemia by genome-wide association study analyses. A single nucleotide polymorphism (SNP) of ARHGEF12 located in an intron predict- ed to be a GATA1 binding site, rs10892563, is significantly associated with patients who need RBC transfusion (P=3.469E-03, odds ratio 5.864). A luciferase reporter assay revealed that this SNP impairs GATA1-mediated trans-regulation of ARHGEF12, and quantitative polymerase chain reaction studies confirmed that the homozygotes status is associated with an approximately 61% reduction in ARHGEF12 expression (P=0.0088). Consequently, erythropoiesis was affected at the pro-erythroblast phases. The role of ARHGEF12 and its homologs in erythroid differentiation was confirmed in human K562 cells, mouse 32D cells and primary murine bone marrow cells. We further demonstrated in zebrafish by morpholino-medi- ated knockdown and CRISPR/Cas9-mediated knockout of arhgef12 that its reduction resulted in erythropoiesis defects. The p38 kinase pathway was affected by the ARHGEF12-RhoA signaling in K562 cells, and consistently, the Arhgef12-RhoA-p38 pathway was also shown to be important for ery- throid differentiation in zebrafish as active RhoA or p38 readily rescued the impaired erythropoiesis caused by arhgef12 knockdown. Finally, ARHGEF12-mediated p38 activity also appeared to be involved in pheno- types of patients of the rs10892563 homozygous genotype. Our findings present a novel SNP of ARHGEF12 that may involve ARHGEF12-RhoA-p38 signaling in erythroid regeneration in ALL patients after chemotherapy.
Introduction
Chemotherapy for hematologic malignancies such as acute lymphoblastic leukemia (ALL) often causes anemia. To alleviate chemotherapy-induced anemia, red blood cell (RBC) transfusion has become standard care. The need for RBC
Ferrata Storti Foundation
Haematologica 2020 Volume 105(4):925-936
Correspondence:
YI ZHENG
Yi.Zheng@cchmc.org
YONG ZHOU
zhouyong@sibs.ac.cn
SHUHONG SHEN
shenshuhong@scmc.com.cn
Received: October 30, 2018. Accepted: August 22, 2019. Pre-published: August 29, 2019.
doi:10.3324/haematol.2018.210286
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/105/4/925
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