The discovery of new and effective treatments for human cardiovascular diseases requires the Identification and validation of novel disease mechanisms. Recently, studies of genomic variation entered a new phase. In which unbiased genome-wide association studies (GWAS) can identify novel genetic loci associated with common diseases. We have recently described 95 loci associate with blood lipid levels-LDL cholesterol (LDL-C), HDL cholesterol (HDL-C), or triglycerides, which are strongly associated with risk for myocardial infarction (Ml) In the NHLBl Framingham Heart Study (FHS) and other population cohorts. Much work will be needed to convert the novel associations into functional insights and, ultimately, therapies to reduce the risk of Ml. A key step is to determine how these genetic loci affect phenotypes in human tissue types relevant to lipid metabolism, principally liver and adipose. We have performed expression quantitative trait locus (eQTL) analyses of genotype vs. gene expression in surgical liver and adipose tissue samples from patients;from this work, we found a strong association between an LDL-C- and Ml-associated SNP on chromosome 1p13-rs12740374-and hepatic expression of the S0RT1 gene. However, these studies were limited by scarcity of tissue and the inability to address key cellular phenotypes such as lipoprotein secretion. Thus, there is a need to establish infinitely renewable sources of hepatocytes and adipocytes from patients of defined genotypes. We have developed the ability to obtain reprogrammed induced pluripotent stem (IPS) cells via peripheral blood cell (PBC) collection. We have also developed a novel technology to accelerate and scale up IPS generation, without any genomic alteration, using in vitro transcribed RNAs encoding reprogramming factors. Finally, we have established protocols to differentiate iPS cells into functional hepatocytes and adipocytes. Our consortium comprises investigators from the FHS, the Harvard Stem Cell Institute (HSCI), Massachusetts General Hospital (MGH), and Harvard Medical School (HMS). We are uniquely positioned to develop a library of IPS cell lines and iPS- derived hepatocytes and adipocytes from several hundred FHS participants, who have genome-wide genotype data as well as many cardiovascular phenotypes. We propose to leverage these resources to: (1) develop efficient protocols to obtain IPS cell lines from blood samples, followed by differentiation into functional hepatocytes and adipocytes;(2) scale up these protocols to enable high-throughput generation of IPS cell lines, hepatocytes, and adipocytes from ~400 Individuals In the Framingham Offspring Cohort;and (3) perform gene expression and metaboiomic profiling from these hepatocytes and adipocytes, enabling Integrative statistical analyses of genotypes with gene expression and metabolite levels, as well as existing phenotype data such as subclinical measures of atherosclerosis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HL107440-02
Application #
8294699
Study Section
Special Emphasis Panel (ZHL1-CSR-N (F1))
Program Officer
Srinivas, Pothur R
Project Start
2011-07-05
Project End
2016-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
2
Fiscal Year
2012
Total Cost
$1,337,440
Indirect Cost
$207,879
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
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Friesen, Max; Camahort, Raymond; Lee, Youn-Kyoung et al. (2017) Activation of IRF1 in Human Adipocytes Leads to Phenotypes Associated with Metabolic Disease. Stem Cell Reports 8:1164-1173
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Lee, Jungmin; Dykstra, Brad; Spencer, Joel A et al. (2017) mRNA-mediated glycoengineering ameliorates deficient homing of human stem cell-derived hematopoietic progenitors. J Clin Invest 127:2433-2437
Gutierrez-Martinez, Paula; Rossi, Derrick J; Beerman, Isabel (2016) DNA Damage and Aging Around the Clock. Trends Mol Med 22:635-7
Hendriks, William T; Warren, Curtis R; Cowan, Chad A (2016) Genome Editing in Human Pluripotent Stem Cells: Approaches, Pitfalls, and Solutions. Cell Stem Cell 18:53-65
Pope, Benjamin D; Warren, Curtis R; Parker, Kevin Kit et al. (2016) Microenvironmental Control of Adipocyte Fate and Function. Trends Cell Biol 26:745-755
Peters, Derek T; Henderson, Christopher A; Warren, Curtis R et al. (2016) Asialoglycoprotein receptor 1 is a specific cell-surface marker for isolating hepatocytes derived from human pluripotent stem cells. Development 143:1475-81
Dykstra, Brad; Lee, Jungmin; Mortensen, Luke J et al. (2016) Glycoengineering of E-Selectin Ligands by Intracellular versus Extracellular Fucosylation Differentially Affects Osteotropism of Human Mesenchymal Stem Cells. Stem Cells 34:2501-2511
Vedantham, Vasanth; Galang, Giselle; Evangelista, Melissa et al. (2015) RNA sequencing of mouse sinoatrial node reveals an upstream regulatory role for Islet-1 in cardiac pacemaker cells. Circ Res 116:797-803

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