PMID- 28714864 OWN - NLM STAT- MEDLINE DCOM- 20171009 LR - 20240210 IS - 1558-8238 (Electronic) IS - 0021-9738 (Print) IS - 0021-9738 (Linking) VI - 127 IP - 8 DP - 2017 Aug 1 TI - An erythroid-specific ATP2B4 enhancer mediates red blood cell hydration and malaria susceptibility. PG - 3065-3074 LID - 94378 [pii] LID - 10.1172/JCI94378 [doi] AB - The lack of mechanistic explanations for many genotype-phenotype associations identified by GWAS precludes thorough assessment of their impact on human health. Here, we conducted an expression quantitative trait locus (eQTL) mapping analysis in erythroblasts and found erythroid-specific eQTLs for ATP2B4, the main calcium ATPase of red blood cells (rbc). The same SNPs were previously associated with mean corpuscular hemoglobin concentration (MCHC) and susceptibility to severe malaria infection. We showed that Atp2b4-/- mice demonstrate increased MCHC, confirming ATP2B4 as the causal gene at this GWAS locus. Using CRISPR-Cas9, we fine mapped the genetic signal to an erythroid-specific enhancer of ATP2B4. Erythroid cells with a deletion of the ATP2B4 enhancer had abnormally high intracellular calcium levels. These results illustrate the power of combined transcriptomic, epigenomic, and genome-editing approaches in characterizing noncoding regulatory elements in phenotype-relevant cells. Our study supports ATP2B4 as a potential target for modulating rbc hydration in erythroid disorders and malaria infection. FAU - Lessard, Samuel AU - Lessard S AD - Montreal Heart Institute and Universite de Montreal, Montreal, Quebec, Canada. FAU - Gatof, Emily Stern AU - Gatof ES AD - Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA. AD - Tufts University School of Medicine, Boston, Massachusetts, USA. FAU - Beaudoin, Melissa AU - Beaudoin M AD - Montreal Heart Institute and Universite de Montreal, Montreal, Quebec, Canada. FAU - Schupp, Patrick G AU - Schupp PG AD - Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA. FAU - Sher, Falak AU - Sher F AD - Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA. FAU - Ali, Adnan AU - Ali A AD - Cancer Research UK Manchester Institute, and. FAU - Prehar, Sukhpal AU - Prehar S AD - Division of Cardiovascular Sciences, The University of Manchester, Manchester Academic Health Science Centre, Manchester, United Kingdom. FAU - Kurita, Ryo AU - Kurita R AD - Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan. FAU - Nakamura, Yukio AU - Nakamura Y AD - Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan. AD - Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan. FAU - Baena, Esther AU - Baena E AD - Cancer Research UK Manchester Institute, and. FAU - Ledoux, Jonathan AU - Ledoux J AD - Montreal Heart Institute and Universite de Montreal, Montreal, Quebec, Canada. FAU - Oceandy, Delvac AU - Oceandy D AD - Division of Cardiovascular Sciences, The University of Manchester, Manchester Academic Health Science Centre, Manchester, United Kingdom. FAU - Bauer, Daniel E AU - Bauer DE AD - Division of Hematology/Oncology, Boston Children's Hospital, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Stem Cell Institute, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA. FAU - Lettre, Guillaume AU - Lettre G AD - Montreal Heart Institute and Universite de Montreal, Montreal, Quebec, Canada. LA - eng GR - K08 DK093705/DK/NIDDK NIH HHS/United States GR - MR/P015816/1/MRC_/Medical Research Council/United Kingdom GR - 19996/CRUK_/Cancer Research UK/United Kingdom GR - MC_QA137853/MRC_/Medical Research Council/United Kingdom GR - R03 DK109232/DK/NIDDK NIH HHS/United States GR - DP2 HL137300/HL/NHLBI NIH HHS/United States GR - PG/13/12/30017/BHF_/British Heart Foundation/United Kingdom GR - PG/16/77/32400/BHF_/British Heart Foundation/United Kingdom PT - Journal Article DEP - 20170717 PL - United States TA - J Clin Invest JT - The Journal of clinical investigation JID - 7802877 RN - 0 (PMCA4 protein, mouse) RN - EC 3.6.1.8 (ATP2B4 protein, human) RN - EC 3.6.3.8 (Plasma Membrane Calcium-Transporting ATPases) RN - EC 7.2.2.10 (Calcium-Transporting ATPases) RN - SY7Q814VUP (Calcium) SB - IM MH - Animals MH - CRISPR-Cas Systems MH - Calcium/metabolism MH - Calcium-Transporting ATPases/*genetics/metabolism MH - Chromosome Mapping MH - Enhancer Elements, Genetic MH - Epigenomics MH - Erythroblasts/metabolism MH - Erythrocytes/*cytology MH - Gene Expression Profiling MH - Gene Regulatory Networks MH - *Genetic Predisposition to Disease MH - Genome-Wide Association Study MH - HEK293 Cells MH - Humans MH - Malaria/*genetics/metabolism MH - Male MH - Mice MH - Mice, Transgenic MH - Phenotype MH - Plasma Membrane Calcium-Transporting ATPases/*genetics/metabolism MH - Polymorphism, Single Nucleotide MH - Quantitative Trait Loci PMC - PMC5531409 COIS- Conflict of interest: The authors have declared that no conflict of interest exists. EDAT- 2017/07/18 06:00 MHDA- 2017/10/11 06:00 PMCR- 2017/11/01 CRDT- 2017/07/18 06:00 PHST- 2017/04/03 00:00 [received] PHST- 2017/06/01 00:00 [accepted] PHST- 2017/07/18 06:00 [pubmed] PHST- 2017/10/11 06:00 [medline] PHST- 2017/07/18 06:00 [entrez] PHST- 2017/11/01 00:00 [pmc-release] AID - 94378 [pii] AID - 10.1172/JCI94378 [doi] PST - ppublish SO - J Clin Invest. 2017 Aug 1;127(8):3065-3074. doi: 10.1172/JCI94378. Epub 2017 Jul 17.