Clonal hematopoiesis of indeterminate potential, DNA methylation, and risk for coronary artery disease

Age-related changes to the genome-wide DNA methylation (DNAm) pattern observed in blood are well-documented. Clonal hematopoiesis of indeterminate potential (CHIP), characterized by the age-related acquisition and expansion of leukemogenic mutations in hematopoietic stem cells (HSCs), is associated with blood cancer and coronary artery disease (CAD). Epigenetic regulators DNMT3A and TET2 are the two most frequently mutated CHIP genes. Here, we present results from an epigenome-wide association study for CHIP in 582 Cardiovascular Health Study (CHS) participants, with replication in 2655 Atherosclerosis Risk in Communities (ARIC) Study participants. We show that DNMT3A and TET2 CHIP have distinct and directionally opposing genome-wide DNAm association patterns consistent with their regulatory roles, albeit both promoting self-renewal of HSCs. Mendelian randomization analyses indicate that a subset of DNAm alterations associated with these two leading CHIP genes may promote the risk for CAD

Mendelian randomization supports bidirectional causality between telomere length and clonal hematopoiesis of indeterminate potential

Human genetic studies support an inverse causal relationship between leukocyte telomere length (LTL) and coronary artery disease (CAD), but directionally mixed effects for LTL and diverse malignancies. Clonal hematopoiesis of indeterminate potential (CHIP), characterized by expansion of hematopoietic cells bearing leukemogenic mutations, predisposes both hematologic malignancy and CAD. TERT (which encodes telomerase reverse transcriptase) is the most significantly associated germline locus for CHIP in genome-wide association studies. Here, we investigated the relationship between CHIP, LTL, and CAD in the Trans-Omics for Precision Medicine (TOPMed) program (n = 63,302) and UK Biobank (n = 47,080). Bidirectional Mendelian randomization studies were consistent with longer genetically imputed LTL increasing propensity to develop CHIP, but CHIP then, in turn, hastens to shorten measured LTL (mLTL). We also demonstrated evidence of modest mediation between CHIP and CAD by mLTL. Our data promote an understanding of potential causal relationships across CHIP and LTL toward prevention of CAD

Amino acid carriers of Ricinus communis expressed during seedling development: molecular cloning and expression analysis of two putative amino acid transporters, RcAAP and RcAAP2

This study reports on the isolation of two putative amino acid carrier cDNAs, RcAAP1 and RcAAP2, from Ricinus communis. Northern analysis shows that RcAAP1 and RcAAP2 are expressed abundantly in the cotyledon and root tissues of developing seedlings and at lower levels in the endosperm and hypocotyl. In the mature plant low expression was observed in the source and sink leaves. We have further characterized the expression of RcAAP1 in Ricinus roots by in situ hybridization. The transcripts are localized in many cell types of the root tip region, including the epidermal and cortical cells, but the highest expression was observed in the cells of the stele situated adjacent to the xylem poles. This is the first report describing the cellular expression of an amino acid transporter in roots, and the results are discussed in relation to the physiological role of this transporte

Expression analysis of sucrose and amino acid carriers in Ricinus communis

Paper given at Plant Biology '97 held in Vancouver, British Columbia, Canada on August 2-6, 1997

Biochemical and molecular characterization of sucrose and amino acid carriers in <i>Ricinus communis</i>

The use of energized plasma membrane vesicle preparations from cotyledons and roots of Ricinus communis seedlings is described, and evidence is presented for the existence of plasma membrane H+/sucrose and H+/amino acid symporters. Using fractions isolated from roots, there is evidence for at least two carriers which can transport neutral amino acids and one which can also transport basic amino acids. A method for the solubilization and reconstitution of glutamine transport activity is described. Preliminary results on the molecular characterization of two putative amino acid carriers and a putative sucrose transporter from Ricinus are presented

Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis

This study describes the expression of a sucrose carrier at various developmental stages in Ricinus communis. A partial-length cDNA clone, RcSUT1, was isolated by RT-PCR from Ricinus seedling RNA. This is almost identical to a sucrose carrier cDNA, Rscr1, which has previously been isolated by library screening. However, we have observed a very different expression pattern in the seedling to that previously reported. Northern analysis, with RcSUT1 as a probe, revealed high expression of a 2 kb transcript in the cotyledons of the germinating seedling; transcript levels were similar in cotyledons harvested 3–6 days after germination. A much lower level of this transcript was detected in the root, hypocotyl and endosperm RNA of the seedling and very low levels were also present in the sink and source leaves of the mature plant. This pattern of expression was also reflected at the protein level with an antipeptide antibody raised to part of the RcSUT1 deduced amino acid sequence.Tissue print hybridisation analysis of the hypocotyl revealed that the sucrose carrier transcripts were localised to the phloem cells of the vascular bundles. A more detailed analysis of sucrose carrier gene expression in the cotyledons of the germinating seedling was carried out by in situ hybridisation; the strongest signals were observed from the lower epidermal layer and the phloem, consistent with an active loading role for these cells. An ultrastructural study of the cells in the lower epidermis showed that they have wall ingrowths which are characteristic of transfer cells. The results are discussed in relation to the physiological role of the sucrose carrier in the Ricinus seedling and to the pathways of sucrose movement from endosperm to the sieve elements in the cotyledon

Cloning of a cDNA coding for an amino acid carrier from Ricinus communis (RcAAP1) by functional complementation in yeast: kinetic analysis, inhibitor sensitivity and substrate specificity

A cDNA for the amino acid permease gene RcAAP1 has been isolated from Ricinus communis by yeast complementationand subjected to a detailed kinetic analysis. RcAAP1 cDNA is 1.5 kb with an open reading frame that codes for a proteinwith 486 amino acids and a calculated molecular mass of 53.1 kDa. RcAAPl-mediated histidine uptake was pH dependentwith highest transport rates at acidic pH; it was sensitive to protonophores and uncouplers and the Km for histidine uptakewas 96 gM. The substrate specificity was investigated by measuring the levels of inhibition of histidine uptake by a range ofamino acids. The basic amino acids (histidine, lysine and arginine) showed strongest inhibition of uptake whereas acidicamino acids competed less effectively. Alanine was the most efficient competitor of the neutral amino acids. Glutamine,serine, asparagine, methionine and cysteine showed moderate inhibition whereas threonine, isoleucine, leucine, phenylalanine, tyrosine and tryptophan showed only low levels of inhibition. Glycine, proline and citrulline caused slightstimulation. More detailed competition kinetics indicated that both lysine and arginine showed simple competitive inhibitionof histidine uptake. When direct uptake measurements were carried out, both lysine and arginine were found to be effectivesubstrates for RcAAP