Elucidation of the mechanism of peritoneal dialysis-related peritoneal injury through complement pathway and establishment of novel anti-complement therapy.

• Project/Area Number: 21K08275
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53040:Nephrology-related
• Research Institution: Nagoya University
• Principal Investigator: 鈴木 康弘 名古屋大学, 医学系研究科, 特任講師 (20584676)
• Co-Investigator(Kenkyū-buntansha): 石本 卓嗣 愛知医科大学, 医学部, 教授 (00534835) ; 水野 正司 名古屋大学, 医学系研究科, 特任教授 (20303638) ; 金 恒秀 名古屋大学, 医学系研究科, 特任助教 (40745238) ; 伊藤 恭彦 愛知医科大学, 経営戦略推進本部, 特命教授 (60402632)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Project Status: Discontinued (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2023: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 人工透析学 / 腹膜透析 / 補体活性化 / 補体制御 / 腹膜傷害 / 腹膜障害 / 補体

Outline of Research at the Start

腹膜透析療法において、補体はいかにしてその活性と制御のバランスを乱し腹膜傷害に進展するのか検討するために、PD患者排液を用いてPD期間による補体系分子発現の変化を比較検討する。PD離脱時腹膜組織の病理学的解析とあわせ、PD療法の経年的腹膜傷害における補体の関与の全体像を明らかにする。腹膜炎時のPD排液、腹膜組織において腹膜炎から進展する腹膜傷害における補体の関与を明らかにする。
また、補体経路でどの経路がより重要であるかを検討するために、補体成分に対する中和抗体やノックアウトマウスを用いて補体経路の要所をブロックすることで最も有効な補体経路のターゲットに対する抗補体療法を確立する。

Outline of Annual Research Achievements

1. 腹膜透析（PD）患者のPD排液から得られる中皮細胞の初代培養細胞（HMPC）を用いて補体活性化を検討した。３つの膜補体制御因子(CReg)、CD46、CD55、CD59について、PD継続期間の前後1年でPCR、FACSを用いて、mRNAと蛋白の発現を観察した。結果は、CD46、CD55、CD59の中で、CD46とCD59の発現が、mRNAレベルと細胞表面の蛋白発現の両方で有意に増加していることを確認した。一方で、CD55の発現には有意な変化を認めなかった。HMPC作成のために回収したPD排液中のsC5b-9レベルも低下していることを示し、一部を論文化した（Front Med. 9.972592, 2022）。
2. 腹膜中皮細胞上のCReg発現の変化が起こる原因について、現在、長期PDが、生体の腹膜に与える影響、特に補体活性系と制御系のバランスに与える影響の検討を継続してすすめている。
3. 腹膜中皮細胞株（Met-5A 細胞）の解析を継続中である。本年度は、1.で得られたHMPC上のCRegの変化について、乳酸、ブドウ糖、pHの影響、特に前者は反復刺激による影響を確認し、一部を論文化した（Front Med. 9.972592, 2022）。
4. 腹膜傷害への治療介入を期待して、現在、これまで未知であった補体の終末経路を抑制する中和抗体を用いて、真菌性腹膜炎の実験動物モデルでの炎症細胞集積、脈管新生、及び線維化進展などの腹膜傷害の進展への補体終末経路の役割の検討を進めている。その結果、中和抗体により有意な腹膜炎症抑制効果が得られ、これをまとめて学会に発表し（EMCHD 2022）、現在投稿中である。

Research Products

• [Journal Article] Recurrent Cerebrovascular Complications under Enzyme Replacement Therapy in a Patient with Fabry Disease on Peritoneal Dialysis (2023)
  • Author(s): Muto R. Suzuki Y, Shimizu H, Yasuda K, Ishimoto T, Maruyama S, Ito Y, Mizuno M.
  • Journal Title: Internal Medicine Volume: 62 Issue: 4 Pages: 565-569
  • DOI: 10.2169/internalmedicine.0185-22
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2023-02-15
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of Transglutaminase 2 Reduces Peritoneal Injury in a Chlorhexidine-Induced Peritoneal Fibrosis Model (2023)
  • Author(s): Kunoki S, Tatsukawa H, Sakai Y, Kinashi H, Kariya T, Suzuki Y, Mizuno M, Yamaguchi M, Sasakura H, Ikeno M, Takeuchi K, Ishimoto T, Hitomi K, Ito Y
  • Journal Title: Laboratory Investigation Volume: 103 Issue: 4 Pages: 100050-100050
  • DOI: 10.1016/j.labinv.2022.100050
  • Peer Reviewed / Open Access
• [Journal Article] Serum and plasma levels of Ba, but not those of soluble C5b-9, might be affected by renal function in chronic kidney disease patients. (2023)
  • Author(s): Yamane R, Yasuda Y, Oshima A, Suzuki Y, Kojima H, Kim H, Fukui S, Maruyama S, Ito Y, Mizuno M.
  • Journal Title: BMC Nephrol. Volume: 24 Issue: 1 Pages: 26-26
  • DOI: 10.1186/s12882-022-03022-z
  • Peer Reviewed / Open Access
• [Journal Article] Long-term peritoneal dialysate exposure modulates expression of membrane complement regulators in human peritoneal mesothelial cells. (2022)
  • Author(s): Kobayashi K, Ozeki T. Kim H, Imai M, Kojima H, Iguchi D, Fukui S, Suzuki M, Suzuki Y, Maruyama S, Ito Y, Mizuno M.
  • Journal Title: Front Med. Volume: 9 Pages: 972592-972592
  • DOI: 10.3389/fmed.2022.972592
  • Peer Reviewed / Open Access
• [Journal Article] Refractory Hypotension Caused by Selenium Deficiency in a Patient on Peritoneal Dialysis (2021)
  • Author(s): Ryuge A, Kim H, Suzuki Y, Okazaki M, Kojima H, Saito S, Kato N, Ishimoto T, Kosugi T, Maruyama S, Mizuno M.
  • Journal Title: Internal Medicine Volume: 60 Issue: 15 Pages: 2461-2464
  • DOI: 10.2169/internalmedicine.6632-20
  • NAID: 130008070597
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2021-08-01
  • Peer Reviewed
• [Journal Article] Oral Antibiotics are Effective for Preventing Colonoscopy-associated Peritonitis as a Preemptive Therapy in Patients on Peritoneal Dialysis (2021)
  • Author(s): Suzuki Y, Mizuno M, Kojima H, Sato Y, Kim H, Kinashi H, Katsuno T, Maruyama S, Ito Y.
  • Journal Title: Internal Medicine Volume: 60 Issue: 3 Pages: 353-356
  • DOI: 10.2169/internalmedicine.5092-20
  • NAID: 130007979708
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2021-02-01
  • Peer Reviewed
• [Journal Article] A case of temporary metastatic pulmonary calcification on peritoneal dialysis accompanied with ectopic hyperparathyroidism. (2021)
  • Author(s): Kawase K, Takagi K, Mizuno M, Horie M.
  • Journal Title: Clin Nephrol. Volume: 95 Issue: 3 Pages: 161-165
  • DOI: 10.5414/cn110337
  • Peer Reviewed
• [Journal Article] C1 inhibitor mitigates peritoneal injury in zymosan-induced peritonitis. (2021)
  • Author(s): Ozeki T, Mizuno M, Iguchi D, Kojima H, Kim H, Suzuki Y, Kinashi H, Ishimoto T, Maruyama S, Ito Y.
  • Journal Title: Am J Physiol Renal Physiol. Volume: － Issue: 6 Pages: F1123-F1132
  • DOI: 10.1152/ajprenal.00600.2020
  • Peer Reviewed
• [Journal Article] Safety and utility of the alpha replacer for treatment of intraluminal obstruction of peritoneal catheters by fibrin clots. (2021)
  • Author(s): Asai A, Kinashi H, Suzuki Y, Kojima H, Sato Y, Matsuoka N, Asai N, Kuroyanagi Y, Yamaguchi M, Nobata H, Katsuno T, Ishimoto T, Mizuno M, Ito Y.
  • Journal Title: Clin Exp Nephrol. Volume: 25 Issue: 4 Pages: 418-427
  • DOI: 10.1007/s10157-020-02001-8
  • Peer Reviewed
• [Journal Article] Low-GDP, pH-neutral solutions preserve peritoneal endothelial glycocalyx during long-term peritoneal dialysis (2021)
  • Author(s): Sugiyama N, Tawada M, Sun T, Suzuki Y, Kinashi H, Yamaguchi M, Katsuno T, Aten J, Vlahu CA, van Kuppevelt TH, Takei Y, Ishimoto T, Maruyama S, Mizuno M, Ito Y.
  • Journal Title: Clin Exp Nephrol Volume: 25 Issue: 9 Pages: 1035-1046
  • DOI: 10.1007/s10157-021-02078-9
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Expression of a Crry/p65 is reduced in acute lung injury induced by extracellular histones (2021)
  • Author(s): Nagano Fumihiko、Mizuno Tomohiro、Imai Masaki、Takahashi Kazuo、Tsuboi Naotake、Maruyama Shoichi、Mizuno Masashi
  • Journal Title: FEBS Open Bio Volume: 12 Issue: 1 Pages: 192-202
  • DOI: 10.1002/2211-5463.13322
  • Peer Reviewed / Open Access
• [Journal Article] A case of C3 glomerulopathy with nephritis-associated plasmin receptor positivity without a history of streptococcal infection (2021)
  • Author(s): Asano Marina、Oda Takashi、Mizuno Masashi
  • Journal Title: CEN Case Reports Volume: 11 Issue: 2 Pages: 259-264
  • DOI: 10.1007/s13730-021-00662-2
  • Peer Reviewed / Open Access
• [Journal Article] Demographic, clinical characteristics and treatment outcomes of immune-complex membranoproliferative glomerulonephritis and C3 glomerulonephritis in Japan: A retrospective analysis of data from the Japan Renal Biopsy Registry (2021)
  • Author(s): Nakagawa Naoki、Mizuno Masashi、Kato Sawako、Maruyama Shoichi、Sato Hiroshi、Nakaya Izaya、Sugiyama Hitoshi、et al.
  • Journal Title: PLOS ONE Volume: 16 Issue: 9 Pages: e0257397-e0257397
  • DOI: 10.1371/journal.pone.0257397
  • Peer Reviewed / Open Access
• [Presentation] Modulation of the complement terminal pathway ameliorates peritoneal injuries in a rat peritoneal dialysis model. (2022)
  • Author(s): Mizuno M, Ozeki T, Iguchi D, Kojima H, Suzuki Y, Kinashi H, Maruyama S, Ito Y.
  • Organizer: EMCHD 2022（国際学会）
  • Int'l Joint Research
• [Presentation] C1 inhibitor prevents peritoneal injuries in a rat peritonitis model. (2022)
  • Author(s): Mizuno M, Ozeki T, Iguchi D, Kojima H, Suzuki Y, Kinashi H, Maruyama S, Ito Y.
  • Organizer: ISPD 2022（国際学会）
  • Int'l Joint Research
• [Presentation] A history of abdominal surgery do not necessary decrease the feasibility of PD: second report on a single-center observational study. (2022)
  • Author(s): Kojima H, Suzuki Y, Yoshioka T, Ishii T, Sakata F, Sato Y, Kinashi H, Katsuno T, Matsukawa Y, Ito Y, Mizuno M.
  • Organizer: ISPD 2022（国際学会）
  • Int'l Joint Research
• [Presentation] Comparison of early withdrawal within three years after induction of peritoneal dialysis (PD) in the Tokai PD registries 1 and 2 in Japan. (2021)
  • Author(s): Mizuno M, Ito Y, Suzuki Y, Kojima H, Saka Y, Morinaga T, Mizutani M, Nonaka K, Naruse N, Hiramatsu T, Ohashi N, Kasuga H, Shimizu H, Kurata H, Kurata K, Suzuki S, Horie M, Maruyama S, Matsuo S
  • Organizer: ISPD EuroPD Joint Congress 2020
  • Int'l Joint Research