KAKENHI (Grants-in-Aid for Scientific Research) -
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International Study for Development and Clinical Aplication of Lupus Biomarkers Reflecting Renal Inflammation
Grant number：17H04667 2017.4 - 2020.3
Tsuboi Naotake
Authorship：Coinvestigator(s)
Because kidney biopsy essential for diagnosis is associated with risk of bleeding and requires hospitalization of patients, repeated examinations for the evaluation of therapeutic efficacy and disease recurrence are not encouraged.
We hypothesized that surface molecules on inflammatory glomerular leukocytes leak into urine. Accordingly, we analyzed bio-samples from domestic and international patients with kidney diseases. The study revealed significant elevation of urinary CD11b with increased numbers of glomerular leukocytes expressing CD11b in patients with lupus nephritis, a major kidney complication of systemic lupus erhythematosus, and in patients with vasculitis. Further analysis of the diagnostic performance for predicting active lupus nephritis demonstrated marked superiority of urinary CD11b compared with two other leukocyte-derived molecules (CD163 and CD16b). -
Elucidation of intracellular import and metabolic circuits involving CD147/Basigin and development of novel therapeutics through regulation of sugar chain modification
Grant number：17K09695 2017.4 - 2020.3
Kosugi Tomoki
Authorship：Coinvestigator(s)
In the present study, we aimed to elucidate the underlying mechanisms of chronic kidney disease (CKD) and hepatic steatosis caused by nutritional overburden more in details. Therefore, we investigated the role of CD147/Basigin (BSG) to maintain intracellular energy homeostasis, using high fat diet (HFD)-induced wild-type and BSG gene-deficient (BSGKO) mice. In the kidneys of BSGKO mice, vacuolar degeneration of tubular epithelial cells and accumulation of lipofuscin-like substances were remarkably suppressed. These findings might be due to the phenomena that transport of the substrates such as pyruvate and lactate in the cells was suppressed by BSG deficits subsequently leading to the the reduction of TCA cycle intermediates and enhanced ketogenesis through activation of free fatty acid beta oxidation. In the setting, augmented BSG protein might be due to the promotion of autophagy, but not translocation to nuclei.
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血管内皮障害を呈する腎疾患におけるSulf2の機能解析と新たな治療戦略の探求
2017.4 - 2019.3
科学研究費補助金 基盤研究(C)
増田 智広
Authorship：Coinvestigator(s)
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血管内皮障害を呈する腎疾患におけるSulf2の機能解析と新たな治療戦略の探求
Grant number：17K09694 2017.4 - 2019.3
増田 智広
Authorship：Coinvestigator(s)
細胞外スルファターゼSulf2は、分子量約130kDaの分泌型酵素で細胞外での膜タンパクに付着している糖鎖の脱硫酸化を担いWnt, BMP, GDNF, FGFというヘパリン結合性因子のシグナル伝達を制御する物質である。
腎臓領域ではSulf1およびSulf2が細胞外基質蓄積を制御し糖尿病性腎症悪化に寄与するとの既報がある。また、ヘパラン硫酸-S-ドメインの蓄積は原線維形成を促進し、腎臓での細胞毒性を促進するという知見が加わった。上記よりSulf2Tgは腎保護に寄与すると仮説を立て私はヒトSulf2全身強制発現マウス(HSulf2Tg)と血管内皮細胞特異的Sulf2強発現 (Tie2-Cre/Sulf2)を用いて、腎機能にどのように関与するか機序解明を目指した。
初年度の計画としてはHSulf2Tgの腎障害惹起時に表現型の確認。腎障害を惹起していない野生型とHSulf2Tgを比較し双方ともに腎機能が正常であった。また、腎障害モデルとして交付申請書に記載した抗基底膜抗体により直接的に糸球体内皮細胞を障害する加速型馬杉腎炎モデルに加えて、アドリアマイシン腎症モデル、糖尿病モデルを作成。いずれのモデルにおいても、HSulf2Tgの方が 尿素窒素・クレアチニン・尿タンパクが低値であった。しかし、Tie2-Cre/Sulf2は野生型と比して尿素窒素・クレアチニン・尿タンパクのいずれも差異は認めなかった。 -
CD147による細胞内輸送・代謝機構の解明と糖鎖修飾調整を介した新規治療法の開発
2017.3 - 2020.3
科学研究費補助金 基盤研究(C)
小杉 智規
Authorship：Coinvestigator(s)
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白血球接着因子とその調節分子の糸球体腎炎における機能解析と細胞移入治療への応用
2016.4 - 2019.3
科学研究費補助金 基盤研究(C)
坪井 直毅
Authorship：Coinvestigator(s)
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ExosomesとマイクロRNAを用いた、安全性の高いオーダーメイド治療の開発
2016.4 - 2019.3
科学研究費補助金 基盤研究(C)
加藤 規利
Authorship：Coinvestigator(s)
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Introduction of preventive and preemptive medicine against chronic antibody-mediated rejection for long-term graft survival
Grant number：16H05465 2016.4 - 2019.3
KOBAYASHI TAKAAKi
Authorship：Coinvestigator(s)
Chronic antibody-mediated rejection is one of major obstacles to long-term graft outcome after kidney transplantation. We focused on preventive or preemptive medicine for de novo DSA production, because anti-rejection treatment has proved ineffective after manifestation of renal dysfunction. CD8+ T cell and B cell-targeting drug sensitivity test could be established. The expression level of CD40L on T cell would be a promising biomarker for antibody production. Potential value of miRNA (142-3p, 1913, 374-5p), mRNA (CIITA, CTLA-4) and donor-derived cell free DNA for preemptive treatment was elucidated. Graft accommodation induced by ABO incompatibility was elicited by up-regulation of CD55/59 and down-regulation of HLA class II through ERK inactivation and mTOR inactivation, respectively. Establishment of T cell indirect recognition assay, analysis of T cell receptor (TCR), and the feasibility of drug-induced graft accommodation should be next challenging subjects.
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Functional analyses of leukocyte integrins and its regulatory molecule in glomerulonephritis and those application for cell transfer therapy
Grant number：16K09611 2016.4 - 2019.3
Tsuboi Naotake
Authorship：Coinvestigator(s)
【Background】PILRα expressed on leukocytes has regulatory functions in leukocyte β2 integrin activation during acute inflammation. Here, we investigated its roles in antibody-mediated glomerular inflammation.【Results】PILRα-/- mice with NTS-GN demonstrated severe glomerular injury. Enhanced glomerular neutrophil accumulation was observed in NTS-GN PILRα-/- only under pre-immunized conditions. PILRα-/- neutrophils exhibited enhanced spreading and adhesion to ICs compared to those in WT cells.【Conclusion】PILRα negatively regulates antibody-mediated neutrophil recruitment, leading to renal injury, by inhibiting Mac-1 integrin activation.
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Research on diagnosis, treatment and epidemiology of membrane nephropathy in the Asia-Pacific region
Grant number：16H05839 2016.4 - 2019.3
maruyama Shoichi
Authorship：Principal investigator
Grant amount：\17940000 （ Direct Cost: \13800000 、 Indirect Cost：\4140000 ）
In order to standardize treatment for membranous nephropathy (MN), we investigated the current status of diagnosis, treatment, and epidemiology of the MN in countries of Asia and the Pacific region including Japan. The medical treatment and patient registration system of MN were very different depending on the country. The prevalence of anti-PLA2R antibody in primary MN patients was reconfirmed in Japan as low as about 50%. On the other hand, the prevalence of PLA2R antibodies in patients in China, Taiwan and Korea were all 80%, equivalent to patients in Western countries. On the contrary, the prevalence of THSD7A antibody was slightly higher in Japan than in other countries. In countries where measurement of autoantibodies cannot be performed, it was concluded that pathological staining of antigens would be the first choice. This study was able to collect a lot of information for the standardization and diffusion of future MN medical care.
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Investigation of safer methods of therapeutic miRNA and exosomes
Grant number：16K09610 2016.4 - 2019.3
Kato Noritoshi
Authorship：Coinvestigator(s)
We investigated the pathophysiological role of exogenously applied microRNA (miRNA) in sepsis-induced multiple organ injury. In vitro, we tested possible miRNAs which suppressed the production of pro-inflammatory cytokines. Of these, miR-146a displayed the highest suppressive effect. Sepsis was induced in mice via cecal ligation and puncture (CLP) and an intravenous injection of a complex of miR-146a-expressing plasmid and polyethyleneimine. Treatment with this complex significantly decreased the level of serum inflammatory cytokines, attenuated organ injury, and led to increased survival from sepsis. miR-146a-expressing plasmid was abundantly distributed in splenic macrophages. CLP mice treated with miR-146a displayed significantly decreased NF-κB activation in the spleen. The collective results support the conclusion that the induction of miR-146a expression in splenic macrophages prevents excessive inflammation and sepsis-induced multiple organ injury.
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Development of novel antihypertensive therapy with tissue protection in renal and cardiovascular diseases through elucidation of MK-EETs blood pressure regulation mechanism
Grant number：16K09609 2016.4 - 2019.3
Kato sawako
Authorship：Coinvestigator(s)
In a system aiming at deep observation of the living body using multiphoton confocal laser microscope A1R MP (Nikon), administration of epoxy eicosatrienoic acids (EETs) inhibitor and adenosine inhibitor, induced the vasoconstrictive action in Midkine (MK) deficient mice. The blood vessels were contracted and blood flow was decreased, as blood pressure increased. The blood flow rate was rising. On the other hand, when a nicotinic acetylcholine receptor inhibitor was administered to evaluate the sympathetic nervous system, blood pressure dropped in MK deficient mice, indicating an increase in blood flow. MK has been shown to regulate renal and blood pressure via a vasodilator.
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RCTを用いたイコサペンタ酸（EPA）による腎保護戦略の確立と作用機序の解明
2016.4 - 2018.3
科学研究費補助金 基盤研究(B)
小林 孝彰
Authorship：Coinvestigator(s)
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Drug discovery of orphan drug for congenital kidney disease using kidney visualization transparent model animal
Grant number：16K15468 2016.4 - 2018.3
MARUYAMA Shoichi
Authorship：Principal investigator
Grant amount：\3510000 （ Direct Cost: \2700000 、 Indirect Cost：\810000 ）
We aimed to develop a novel drug discovery platform for medicine for congenital renal disease which was based on in vivo phenotype screening using kidney-visualized-transparent-zebrafish.
As a result, we bred a new line of transparent zebrafish that is more growth efficiency and more transparency than conventional lines. Although, the gene editing-based congenital nephrotic syndrome model lines could not be generated during study period, the congenital whole-body calcification model transparent kidney visualized line was generated using a phosphorus transporter mutant line. As a result of a compound administration experiment using frying fish, it was possible to observe dose-dependent occurrence of edema, kidney malformation, disappearance of nephron, etc. according to the nephrotoxicity of the compound. These data demonstrated the feasibility of in vivo screening system using kidney visualized transparent zebrafish. -
アジア太平洋地域における膜性腎症の診断・治療・疫学に関する調査研究
2016.4 - 2017.3
科学研究費補助金 基盤研究(B)
丸山 彰一
Authorship：Principal investigator
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腎臓可視化透明モデル動物を用いた先天性腎疾患に対するオーファンドラッグの創薬
2016.4 - 2017.3
科学研究費補助金 挑戦的萌芽研究
丸山 彰一
Authorship：Principal investigator
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Study on the clinical and pathological features of PLA2R-related membranous nephropathy in Japan
Grant number：15K09257 2015.4 - 2019.3
Akiyama Shin'ichi
Authorship：Coinvestigator(s)
We studied on the clinical reality and pathophysiology of PLA2R-associated membranous nephropathy, a new concept of primary membranous nephropathy, in Japanese patients. We optimized the method of measuring PLA2R antibody according to the clinical situation of Japanese patients. The PLA2R antibody measurement at the time of renal biopsy was shown not only useful for differentiation of PLA2R-related membranous nephropathy but also useful for predicting prognosis. PLA2R antibody-positive patients who have a history of smoking have been shown to delay complete remission. Patients with a PLA2R antibody concentration of 50 RU/ml or more at the time of renal biopsy were shown to have a significantly worse renal prognosis.
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Treatment for kidney disease using fetal membrane-derived mesenchymal stem cells.
Grant number：15K09256 2015.4 - 2019.3
Katsuno Takayuki
Authorship：Coinvestigator(s)
Applicants' previous studies have identified mesenchymal stem cells (MSC) differentiation under conditions where the serum concentration of the culture solution of fetal membrane-derived MSC (FM-MSC) was changed to 2%. However, in the low serum culture method with a serum concentration of 2%, the proliferation ability of the FN-MSCs was attenuated, and the proliferation rate was decreased in passage3. When hFGF (human fibroblast growth factor) was added for the purpose of promoting the growth of MSCs and the culture medium was introduced, an improvement in the growth rate was obtained but the problem that the undifferentiated state was not maintained became clear. The FM- MSC could not give the same result as that of adipose tissue-derived MSC. This study suggested that low serum culture may be dependent on cell source.
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慢性腎臓病・高血圧におけるフルクトース代謝の役割と分子機構の解明
2015.4 - 2018.3
科学研究費補助金 基盤研究(C)
石本 卓嗣
Authorship：Coinvestigator(s)
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日本における抗PLA2R抗体関連膜性腎症の実態と病態機序の解明
2015.4 - 2018.3
科学研究費補助金 基盤研究(C)
秋山 真一
Authorship：Coinvestigator(s)