Subtype-specific gout susceptibility loci and enrichment of selection pressure on ABCG2 and ALDH2 identified by subtype genome-wide meta-analyses of clinically defined gout patients

Objectives Genome-wide meta-analyses of clinically defined gout were performed to identify subtype-specific susceptibility loci. Evaluation using selection pressure analysis with these loci was also conducted to investigate genetic risks characteristic of the Japanese population over the last 2000–3000 years. Methods Two genome-wide association studies (GWASs) of 3053 clinically defined gout cases and 4554 controls from Japanese males were performed using the Japonica Array and Illumina Array platforms. About 7.2 million single-nucleotide polymorphisms were meta-analysed after imputation. Patients were then divided into four clinical subtypes (the renal underexcretion type, renal overload type, combined type and normal type), and meta-analyses were conducted in the same manner. Selection pressure analyses using singleton density score were also performed on each subtype. Results In addition to the eight loci we reported previously, two novel loci, PIBF1 and ACSM2B, were identified at a genome-wide significance level (p<5.0×10–8) from a GWAS meta-analysis of all gout patients, and other two novel intergenic loci, CD2-PTGFRN and SLC28A3-NTRK2, from normal type gout patients. Subtype-dependent patterns of Manhattan plots were observed with subtype GWASs of gout patients, indicating that these subtype-specific loci suggest differences in pathophysiology along patients’ gout subtypes. Selection pressure analysis revealed significant enrichment of selection pressure on ABCG2 in addition to ALDH2 loci for all subtypes except for normal type gout. Conclusions Our findings on subtype GWAS meta-analyses and selection pressure analysis of gout will assist elucidation of the subtype-dependent molecular targets and evolutionary involvement among genotype, phenotype and subtype-specific tailor-made medicine/prevention of gout and hyperuricaemia

<所内学術研究成果報告>H. 「環境保全・地球環境温暖化防止をターゲットとする新パルプ資源ケナフの栽培と利用に関する研究」

本研究は, エコマテリアルとしての非木材繊維資源に最も適切である一年生植物ケナフ(Hibiscus cannabinus L.)の栽培とその利用を目的に, 1993年より開始した研究である。従来の成果は, すでに本年報1992,\u2794,\u2795,\u2796,\u2797,\u2798,および\u2799年に報告した。特に従来のケナフ栽培の成果の総決算として, 1998年より平塚市および平塚ケナフ普及協会との共同研究が行われてきた。特に, 平塚市では休耕田対策としてケナフの栽培を推奨し, 現在, 栽培したケナフのパルプ化と紙製造を行って市政に還元している。この現状はさらに展開し, 平塚市のみならず日本全国にその輪が広がり大きな活動となっている。これらの栽培や利用は最も基礎的な指導と, より学術的な研究成果の提供が常に必要であり, この点を最も重要な課題としている。さらに, 環境教育に対する展開を学校, 公民館などを中心に行い, 2000年度は, 平塚キャンパスで市内6小学校の生徒28名のケナフ教育を行った。まお, 研究室内では, 栽培研究の他に, a)種子の発芽阻害実験, b)海水による阻害実験, c)生長に伴うクロロフィル量および水分量の測定実験, d)光合成測定実験, e)花の成分(色素)研究, f)葉など各器官の成分研究などを行っている。取り扱った種類も, ローゼル(H. sabdariffa L.)類も加えると30種に近い

Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles.

Nanoparticles represent promising gene delivery systems in biomedicine to facilitate prolonged gene expression with low toxicity compared to viral vectors. Specifically, nanoparticles of calcium phosphate (nCaP), the main inorganic component of human bone, exhibit high biocompatibility and good biodegradability and have been reported to have high affinity for protein or DNA, having thus been used as gene transfer vectors. On the other hand, Octa-arginine (R8), which has a high permeability to cell membrane, has been reported to improve intracellular delivery systems. Here, we present an optimized method for nCaP-mediated gene delivery using an octa-arginine (R8)-functionalized nCaP vector containing a marker or functional gene construct. nCaP particle size was between 220-580 nm in diameter and all R8-functionalized nCaPs carried a positive charge. R8 concentration significantly improved nCaP transfection efficiency with high cell compatibility in human mesenchymal stem cells (hMSC) and human osteoblasts (hOB) in particular, suggesting nCaPs as a good option for non-viral vector gene delivery. Furthermore, pre-treatment with different endocytosis inhibitors identified that the endocytic pathway differed among cell lines and functionalized nanoparticles, with amiloride increasing transfection efficiency of R8-functionalized nCaPs in hMSC and hOB

PIH1D1 interacts with mTOR complex 1 and enhances ribosome RNA transcription

AbstractPIH1D1 is the defining component of the R2TP complex. Recently, R2TP has been reported to stabilize mTOR (mammalian target of rapamycin), an important regulator of cell growth and protein synthesis. Two complexes of mTOR, mTORC1 and mTORC2, have been identified. We demonstrate that immunoprecipitation (IP) of PIH1D1 results in the co-IP of Raptor (mTORC1 specific), but not Rictor (mTORC2 specific), and that knockdown of PIH1D1 decreases mTORC1 assembly, S6 kinase phosphorylation (indicator of mTORC1 activity), and rRNA transcription without affecting mTORC2 in human breast cancer MCF-7 cells. In addition, we provide evidence that PIH1D1 is overexpressed in various breast cancer cell lines. These findings collectively suggest that PIH1D1 may have an important role in mTORC1 regulation in breast cancers

Exosome-bound WD repeat protein Monad inhibits breast cancer cell invasion by degrading amphiregulin mRNA.

Increased stabilization of mRNA coding for key cancer genes can contribute to invasiveness. This is achieved by down-regulation of exosome cofactors, which bind to 3'-UTR in cancer-related genes. Here, we identified amphiregulin, an EGFR ligand, as a target of WD repeat protein Monad, a component of R2TP/prefoldin-like complex, in MDA-MB-231 breast cancer cells. Monad specifically interacted with both the 3'-UTR of amphiregulin mRNA and the RNA degrading exosome, and enhanced decay of amphiregulin transcripts. Knockdown of Monad increased invasion and this effect was abolished with anti-amphiregulin neutralizing antibody. These results suggest that Monad could prevent amphiregulin-mediated invasion by degrading amphiregulin mRNA

Transfection efficiency (TE) and cell viability (CV) in HeLa, Saos-2, hMSC, and hOB cells.

<p>Transfection efficiency (TE) and cell viability (CV) in HeLa, Saos-2, hMSC, and hOB cells.</p

BMP-2 concentration percentages in the cell culture medium of cells pre-treated with endocytosis inhibitors.

<p><b>A)</b> HeLa, <b>b)</b> Saos-2, <b>c)</b> hMSC, and <b>d)</b> hOB cells transfected with CaP nanoparticles loaded with pUC57 plasmid and functionalized with different concentrated solutions of R8 (0.1, 1, 5, 10, 50, 100 mg ml⁻¹), PEI, and protamine. pUC57 group are single shell non-functionalized CaP nanoparticles. The bars represent the mean ± standard deviation. *p < 0.05 compared to transfection of non-pre-treated cells (NT).</p