title: Integrative DNA, RNA, and protein evidence connects TREML4 to coronary artery calcification.
abstract: Coronary artery calcification (CAC) is a heritable and definitive morphologic marker of atherosclerosis that strongly predicts risk for future cardiovascular events. To search for genes involved in CAC, we used an integrative transcriptomic, genomic, and protein expression strategy by using next-generation DNA sequencing in the discovery phase with follow-up studies using traditional molecular biology and histopathology techniques. RNA sequencing of peripheral blood from a discovery set of CAC cases and controls was used to identify dysregulated genes, which were validated by ClinSeq and Framingham Heart Study data. Only a single gene, TREML4, was upregulated in CAC cases in both studies. Further examination showed that rs2803496 was a TREML4 cis-eQTL and that the minor allele at this locus conferred up to a 6.5-fold increased relative risk of CAC. We characterized human TREML4 and demonstrated by immunohistochemical techniques that it is localized in macrophages surrounding the necrotic core of coronary plaques complicated by calcification (but not in arteries with less advanced disease). Finally, we determined by von Kossa staining that TREML4 colocalizes with areas of microcalcification within coronary plaques. Overall, we present integrative RNA, DNA, and protein evidence implicating TREML4 in coronary artery calcification. Our findings connect multimodal genomics data with a commonly used clinical marker of cardiovascular disease.
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
authors: Shurjo K
authors: SK
authors: Sen
authors: National Cancer Institute, NIH, Frederick, MD 21702, USA.
authors: Kimberly C
authors: KC
authors: Boelte
authors: Center for Information Technology, NIH, Bethesda, MD 20892, USA.
authors: Jennifer J
authors: JJ
authors: Barb
authors: Center for Information Technology, NIH, Bethesda, MD 20892, USA.
authors: Roby
authors: R
authors: Joehanes
authors: CVPath Institute, Gaithersburg, MD 20878, USA.
authors: XiaoQing
authors: X
authors: Zhao
authors: CVPath Institute, Gaithersburg, MD 20878, USA.
authors: Qi
authors: Q
authors: Cheng
authors: CVPath Institute, Gaithersburg, MD 20878, USA.
authors: Lila
authors: L
authors: Adams
authors: Moffitt Cancer Center, Tampa, FL 33612, USA.
authors: Jamie K
authors: JK
authors: Teer
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
authors: David S
authors: DS
authors: Accame
authors: Center for Biologics Evaluation and Research, FDA, Bethesda, MD 20892, USA.
authors: Soma
authors: S
authors: Chowdhury
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
authors: Larry N
authors: LN
authors: Singh
authors: Netherlands Genomics-Initiative-Sponsored Netherlands Consortium for Healthy Aging and Department of Epidemiology, Erasmus University Medical Center, 3000 CA Rotterdam, the Netherlands.
authors: Maryam
authors: M
authors: Kavousi
authors: Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, MI 48104, USA.
authors: Patricia A
authors: PA
authors: Peyser
authors: National Cancer Institute, NIH, Frederick, MD 21702, USA.
authors: Laura
authors: L
authors: Quigley
authors: Applied/Developmental Research Directorate, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors: Debra Long
authors: DL
authors: Priel
authors: Applied/Developmental Research Directorate, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors: Karen
authors: K
authors: Lau
authors: Applied/Developmental Research Directorate, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors: Douglas B
authors: DB
authors: Kuhns
authors: National Cancer Institute, NIH, Frederick, MD 21702, USA.
authors: Teizo
authors: T
authors: Yoshimura
authors: Cardiovascular Epidemiology and Human Genomics Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892, USA; National Heart, Lung and Blood Institute's Framingham Heart Study, Framingham, MA 01702, USA.
authors: Andrew D
authors: AD
authors: Johnson
authors: Cardiovascular Epidemiology and Human Genomics Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892, USA; National Heart, Lung and Blood Institute's Framingham Heart Study, Framingham, MA 01702, USA.
authors: Shih-Jen
authors: SJ
authors: Hwang
authors: Cardiovascular and Pulmonary Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892, USA.
authors: Marcus Y
authors: MY
authors: Chen
authors: Cardiovascular and Pulmonary Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892, USA.
authors: Andrew E
authors: AE
authors: Arai
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
authors: Eric D
authors: ED
authors: Green
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA.
authors: James C
authors: JC
authors: Mullikin
authors: CVPath Institute, Gaithersburg, MD 20878, USA.
authors: Frank D
authors: FD
authors: Kolodgie
authors: Cardiovascular Epidemiology and Human Genomics Branch, Division of Intramural Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD 20892, USA; National Heart, Lung and Blood Institute's Framingham Heart Study, Framingham, MA 01702, USA.
authors: Christopher J
authors: CJ
authors: O'Donnell
authors: CVPath Institute, Gaithersburg, MD 20878, USA.
authors: Renu
authors: R
authors: Virmani
authors: Center for Information Technology, NIH, Bethesda, MD 20892, USA.
authors: Peter J
authors: PJ
authors: Munson
authors: National Cancer Institute, NIH, Frederick, MD 21702, USA.
authors: Daniel W
authors: DW
authors: McVicar
authors: National Human Genome Research Institute, NIH, Bethesda, MD 20892, USA. Electronic address: lesb@mail.nih.gov.
authors: Leslie G
authors: LG
authors: Biesecker
mesh_terms: D001483:Base Sequence; D002114:Calcinosis; D003331:Coronary Vessels / Q000473:pathology*; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers; D057809:HEK293 Cells; D006801:Humans; D011506:Proteins / Q000378:metabolism*; D040641:Quantitative Trait Loci; D012313:RNA / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000235:genetics / Q000502:physiology*
chemical_list: D017931:DNA Primers; D011506:Proteins; D011971:Receptors, Immunologic; C000591058:TREML4 protein, human; D012313:RNA; D004247:DNA