title: In vivo regulation of the heme oxygenase-1 gene in humanized transgenic mice.
abstract: Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, producing equimolar amounts of carbon monoxide, iron, and biliverdin. Induction of HO-1 is a beneficial response to tissue injury in diverse animal models of diseases including acute kidney injury. In vitro analysis has shown that the human HO-1 gene is transcriptionally regulated by changes in chromatin conformation, but whether such control occurs in vivo is not known. To enable such an analysis, we generated transgenic mice, harboring an 87-kb bacterial artificial chromosome expressing human HO-1 mRNA and protein and bred these mice with HO-1 knockout mice to generate humanized BAC transgenic mice. This successfully rescued the phenotype of the knockout mice including reduced birth rates, tissue iron overload, splenomegaly, anemia, leukocytosis, dendritic cell abnormalities, and survival after acute kidney injury induced by rhabdomyolysis or cisplatin nephrotoxicity. Transcription factors such as USF1/2, JunB, Sp1, and CTCF were found to associate with regulatory regions of the human HO-1 gene in the kidney following rhabdomyolysis. Chromosome conformation capture and ChIP-loop assays confirmed this in the formation of chromatin looping in vivo. Thus, these bacterial artificial chromosome humanized HO-1 mice are a valuable model to study the human HO-1 gene, providing insight to the in vivo architecture of the gene in acute kidney injury and other diseases.
authors.affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
authors.forename: Junghyun
authors.initials: J
authors.lastname: Kim
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Zarjou
authors.forename: Amie M
authors.initials: AM
authors.lastname: Traylor
authors.forename: Subhashini
authors.initials: S
authors.lastname: Bolisetty
authors.forename: Edgar A
authors.initials: EA
authors.lastname: Jaimes
authors.forename: Travis D
authors.initials: TD
authors.lastname: Hull
authors.forename: James F
authors.initials: JF
authors.lastname: George
authors.forename: Fady M
authors.initials: FM
authors.lastname: Mikhail
authors.forename: Anupam
authors.initials: A
authors.lastname: Agarwal
mesh_terms: D058186:Acute Kidney Injury / Q000201:enzymology / Q000209:etiology; D000818:Animals; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D002945:Cisplatin / Q000633:toxicity; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D051547:Heme Oxygenase-1 / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012206:Rhabdomyolysis / Q000150:complications; D013045:Species Specificity; D013449:Sulfonamides; D014157:Transcription Factors / Q000378:metabolism; D014529:Uridine / Q000031:analogs & derivatives
chemical_list: D002843:Chromatin; D008565:Membrane Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013449:Sulfonamides; D014157:Transcription Factors; C504402:penoxsulam; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D002945:Cisplatin; D014529:Uridine