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Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells.
Autophagy. 2020 10; 16(10):1889-1904.A

Abstract

Macroautophagy/autophagy is a lysosomal degradation system which plays a protective role against kidney injury. RUBCN/Rubicon (RUN domain and cysteine-rich domain containing, Beclin 1-interacting protein) inhibits the fusion of autophagosomes and lysosomes. However, its physiological role in kidney proximal tubular epithelial cells (PTECs) remains uncertain. In the current study, we analyzed the phenotype of newly generated PTEC-specific rubcn-deficient (KO) mice. Additionally, we investigated the role of RUBCN in lipid metabolism using isolated rubcn-deficient PTECs. Although KO mice exhibited sustained high autophagic flux in PTECs, they were not protected from acute ischemic kidney injury. Unexpectedly, KO mice exhibited hallmark features of metabolic syndrome accompanied by expanded lysosomes containing multi-layered phospholipids in PTECs. RUBCN deficiency in cultured PTECs promoted the mobilization of phospholipids from cellular membranes to lysosomes via enhanced autophagy. Treatment of KO PTECs with oleic acid accelerated fatty acids transfer to mitochondria. Furthermore, KO PTECs promoted massive triglyceride accumulation in hepatocytes (BNL-CL2 cells) co-cultured in transwell, suggesting accelerated fatty acids efflux from the PTECs contributes to the metabolic syndrome in KO mice. This study shows that sustained high autophagic flux by RUBCN deficiency in PTECs leads to metabolic syndrome concomitantly with an accelerated mobilization of phospholipids from cellular membranes to lysosomes. Abbreviations: ABC: ATP binding cassette; ACADM: acyl-CoA dehydrogenase medium chain; ACTB: actin, beta; ATG: autophagy related; AUC: area under the curve; Baf: bafilomycin A1; BAT: brown adipose tissue; BODIPY: boron-dipyrromethene; BSA: bovine serum albumin; BW: body weight; CAT: chloramphenicol acetyltransferase; CM: complete medium; CPT1A: carnitine palmitoyltransferase 1a, liver; CQ: chloroquine; CTRL: control; EGFP: enhanced green fluorescent protein; CTSD: cathepsin D; EAT: epididymal adipose tissue; EGFR: epidermal growth factor receptor; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; FA: fatty acid; FBS: fetal bovine serum; GTT: glucose tolerance test; HE: hematoxylin and eosin; HFD: high-fat diet; I/R: ischemia-reperfusion; ITT: insulin tolerance test; KAP: kidney androgen regulated protein; KO: knockout; LAMP1: lysosomal associated membrane protein 1; LD: lipid droplet; LRP2: low density lipoprotein receptor related protein 2; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MAT: mesenteric adipose tissue; MS: mass spectrometry; MTOR: mechanistic target of rapamycin kinase; MTORC1: MTOR complex 1; NDRG1: N-myc downstream regulated 1; NDUFB5: NADH:ubiquinone oxidoreductase subunit B5; NEFA: non-esterified fatty acid; OA: oleic acid; OCT: optimal cutting temperature; ORO: Oil Red O; PAS: Periodic-acid Schiff; PFA: paraformaldehyde; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PPARA: peroxisome proliferator activated receptor alpha; PPARGC1A: PPARG coactivator 1 alpha; PTEC: proximal tubular epithelial cell; RAB7A: RAB7A, member RAS oncogene family; RPS6: ribosomal protein S6; RPS6KB1: ribosomal protein S6 kinase B1; RT: reverse transcription; RUBCN: rubicon autophagy regulator; SAT: subcutaneous adipose tissue; SFC: supercritical fluid chromatography; SQSTM1: sequestosome 1; SREBF1: sterol regulatory element binding transcription factor 1; SV-40: simian virus-40; TFEB: transcription factor EB; TG: triglyceride; TS: tissue specific; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; UN: urea nitrogen; UQCRB: ubiquinol-cytochrome c reductase binding protein; UVRAG: UV radiation resistance associated; VPS: vacuolar protein sorting; WAT: white adipose tissue.

Authors+Show Affiliations

Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan. Reverse Translational Project, Center for Rare Disease Research, National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN) , Osaka, Japan.Department of Advanced Technology for Transplantation, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University , Fukuoka, Japan.Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University , Fukuoka, Japan.Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University , Fukuoka, Japan.Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University , Fukuoka, Japan.Institute of Medical Science and Department of Basic Sciences, Tokai University School of Medicine , Isehara, Japan.Department of Pediatrics, Tokai University School of Medicine , Isehara, Japan.Department of Nephrology, Kyoto University Graduate School of Medicine , Kyoto, Japan. Institute for the Advanced Study of Human Biology (ASHBi), Kyoto University , Kyoto, Japan.Department of Genetics, Osaka University Graduate School of Medicine , Osaka, Japan.Department of Nephrology, Osaka University Graduate School of Medicine , Osaka, Japan.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

31944172

Citation

Matsuda, Jun, et al. "Metabolic Effects of RUBCN/Rubicon Deficiency in Kidney Proximal Tubular Epithelial Cells." Autophagy, vol. 16, no. 10, 2020, pp. 1889-1904.
Matsuda J, Takahashi A, Takabatake Y, et al. Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells. Autophagy. 2020;16(10):1889-1904.
Matsuda, J., Takahashi, A., Takabatake, Y., Sakai, S., Minami, S., Yamamoto, T., Fujimura, R., Namba-Hamano, T., Yonishi, H., Nakamura, J., Kimura, T., Kaimori, J. Y., Matsui, I., Takahashi, M., Nakao, M., Izumi, Y., Bamba, T., Matsusaka, T., Niimura, F., ... Isaka, Y. (2020). Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells. Autophagy, 16(10), 1889-1904. https://doi.org/10.1080/15548627.2020.1712107
Matsuda J, et al. Metabolic Effects of RUBCN/Rubicon Deficiency in Kidney Proximal Tubular Epithelial Cells. Autophagy. 2020;16(10):1889-1904. PubMed PMID: 31944172.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells. AU - Matsuda,Jun, AU - Takahashi,Atsushi, AU - Takabatake,Yoshitsugu, AU - Sakai,Shinsuke, AU - Minami,Satoshi, AU - Yamamoto,Takeshi, AU - Fujimura,Ryuta, AU - Namba-Hamano,Tomoko, AU - Yonishi,Hiroaki, AU - Nakamura,Jun, AU - Kimura,Tomonori, AU - Kaimori,Jun-Ya, AU - Matsui,Isao, AU - Takahashi,Masatomo, AU - Nakao,Motonao, AU - Izumi,Yoshihiro, AU - Bamba,Takeshi, AU - Matsusaka,Taiji, AU - Niimura,Fumio, AU - Yanagita,Motoko, AU - Yoshimori,Tamotsu, AU - Isaka,Yoshitaka, Y1 - 2020/01/16/ PY - 2020/1/17/pubmed PY - 2021/9/28/medline PY - 2020/1/17/entrez KW - Autophagic flux KW - RUBCN/Rubicon KW - autophagy KW - lipid efflux KW - lysosome KW - metabolic syndrome SP - 1889 EP - 1904 JF - Autophagy JO - Autophagy VL - 16 IS - 10 N2 - Macroautophagy/autophagy is a lysosomal degradation system which plays a protective role against kidney injury. RUBCN/Rubicon (RUN domain and cysteine-rich domain containing, Beclin 1-interacting protein) inhibits the fusion of autophagosomes and lysosomes. However, its physiological role in kidney proximal tubular epithelial cells (PTECs) remains uncertain. In the current study, we analyzed the phenotype of newly generated PTEC-specific rubcn-deficient (KO) mice. Additionally, we investigated the role of RUBCN in lipid metabolism using isolated rubcn-deficient PTECs. Although KO mice exhibited sustained high autophagic flux in PTECs, they were not protected from acute ischemic kidney injury. Unexpectedly, KO mice exhibited hallmark features of metabolic syndrome accompanied by expanded lysosomes containing multi-layered phospholipids in PTECs. RUBCN deficiency in cultured PTECs promoted the mobilization of phospholipids from cellular membranes to lysosomes via enhanced autophagy. Treatment of KO PTECs with oleic acid accelerated fatty acids transfer to mitochondria. Furthermore, KO PTECs promoted massive triglyceride accumulation in hepatocytes (BNL-CL2 cells) co-cultured in transwell, suggesting accelerated fatty acids efflux from the PTECs contributes to the metabolic syndrome in KO mice. This study shows that sustained high autophagic flux by RUBCN deficiency in PTECs leads to metabolic syndrome concomitantly with an accelerated mobilization of phospholipids from cellular membranes to lysosomes. Abbreviations: ABC: ATP binding cassette; ACADM: acyl-CoA dehydrogenase medium chain; ACTB: actin, beta; ATG: autophagy related; AUC: area under the curve; Baf: bafilomycin A1; BAT: brown adipose tissue; BODIPY: boron-dipyrromethene; BSA: bovine serum albumin; BW: body weight; CAT: chloramphenicol acetyltransferase; CM: complete medium; CPT1A: carnitine palmitoyltransferase 1a, liver; CQ: chloroquine; CTRL: control; EGFP: enhanced green fluorescent protein; CTSD: cathepsin D; EAT: epididymal adipose tissue; EGFR: epidermal growth factor receptor; EIF4EBP1: eukaryotic translation initiation factor 4E binding protein 1; FA: fatty acid; FBS: fetal bovine serum; GTT: glucose tolerance test; HE: hematoxylin and eosin; HFD: high-fat diet; I/R: ischemia-reperfusion; ITT: insulin tolerance test; KAP: kidney androgen regulated protein; KO: knockout; LAMP1: lysosomal associated membrane protein 1; LD: lipid droplet; LRP2: low density lipoprotein receptor related protein 2; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MAT: mesenteric adipose tissue; MS: mass spectrometry; MTOR: mechanistic target of rapamycin kinase; MTORC1: MTOR complex 1; NDRG1: N-myc downstream regulated 1; NDUFB5: NADH:ubiquinone oxidoreductase subunit B5; NEFA: non-esterified fatty acid; OA: oleic acid; OCT: optimal cutting temperature; ORO: Oil Red O; PAS: Periodic-acid Schiff; PFA: paraformaldehyde; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PPARA: peroxisome proliferator activated receptor alpha; PPARGC1A: PPARG coactivator 1 alpha; PTEC: proximal tubular epithelial cell; RAB7A: RAB7A, member RAS oncogene family; RPS6: ribosomal protein S6; RPS6KB1: ribosomal protein S6 kinase B1; RT: reverse transcription; RUBCN: rubicon autophagy regulator; SAT: subcutaneous adipose tissue; SFC: supercritical fluid chromatography; SQSTM1: sequestosome 1; SREBF1: sterol regulatory element binding transcription factor 1; SV-40: simian virus-40; TFEB: transcription factor EB; TG: triglyceride; TS: tissue specific; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; UN: urea nitrogen; UQCRB: ubiquinol-cytochrome c reductase binding protein; UVRAG: UV radiation resistance associated; VPS: vacuolar protein sorting; WAT: white adipose tissue. SN - 1554-8635 UR - https://www.unboundmedicine.com/medline/citation/31944172/Metabolic_effects_of_RUBCN/Rubicon_deficiency_in_kidney_proximal_tubular_epithelial_cells_ DB - PRIME DP - Unbound Medicine ER -