Center for Molecular Medicine Cologne

Brinkkötter, Paul T | Brähler, Sebastian - B 01

Metabolic reprogramming of renal immune cells in a model of glomerulonephritis

Introduction

Inflammatory diseases of the kidney filter are leading causes of chronic renal insufficiency. Despite tremendous advances in our disease understanding, the treatment options remain limited. Targeted strategies to tackle specific intracellular pathways are warranted. Metabolic reprogramming is one the key steps in immune cells activation. Our project will not only advance our knowledge about a fundamental disease mechanism with the perspective of new treatment approaches for renal diseases.

The kidney contains a complex network of immune cells important in the defence against pathogens and also in development and tissue homeostasis in states of health and disease. The prominent role of inflammatory processes is reflected by the wide use of anti-inflammatory drugs in the treatment of patients with glomerular diseases of many different causes. Metabolic reprogramming has recently been recognized as central event in the regulation of T cell and macrophage function. With respect to the kidney, very little is known about the inflammatory signal transduction itself and how cells of the immune system are orchestrated during glomerular disease. Our overarching goal is to gain mechanistic insights into the role of metabolic signalling events and their influence on myeloid cell function in glomerulonephritis. For this purpose, we perform detailed metabolic profiling of FACS sorted immune cells of the kidney in a model of experimental glomerulonephritis. We are particularly interested in the role of insulin-receptor signalling in the activation of various immune cell subsets in inflammatory kidney diseases.

In this project we will also analyse the role of the tricarboxylic acid (TCA) cycle during inflammatory macrophage activation by studying the function of TCA-cycle associated proteins and their metabolites.

Our ultimate goal is to identify new therapeutic targets and transfer these findings in innovative treatment strategies for our patients.

Our Aims

  1. Analysis of metabolic properties of key immune cell populations during glomerulonephritis.
  2. Characterization of the role of insulin signaling in macrophages, DCs and T-cells.
  3. Evaluation of the function of the TCA cycle and associated metabolites in inflammatory kidney disease.

Own Previous Work

Our lab is interested in the metabolic and inflammatory pathways that lead to kidney failure.

Recently, using innovative imaging techniques like multiphoton-micorscopy were able to redefine the morphology of podocytes, macrophages and two major DC subsets in the mouse kidney: cDC1 and cDC2 (1,2).We challenged mice with the standard model of immune complex-mediated nephritis, i.e. nephrotoxic nephritis, a disease mechanism that causes the most devastating form of GN: crescentic or rapid-progressive GN. We found that the known extensive network of immune cells in the kidney under healthy conditions is not consisting of dendritic cells, but of macrophages, while cDC1 and cDC2 are smaller migratory populations located in clusters (Figure 1).  

Figure 1

We were able to characterize the function of cDC1 and cDC2. cDC2, which represent about 90% of DCs in the kidney, have a strong pro-inflammatory function during GN by recruiting Th17 through IL23 and secreting neutrophil-attracting cytokines. In contrast, the cDC1 subset had a robust anti-inflammatory function by counteracting the activity of the cDC2.

In order to understand the function of cDC1 and cDC2 in detail, we obtained comprehensive gene expression data by RNAseq from sorted mouse kidney cDCs under healthy and inflamed conditions. Our data showed fundamental differences in the gene expression and metabolic profiles of the two DC subsets, supporting the hypothesis that the opposing functions of these immune cell subsets are reflected by their metabolic profile demanding for a comprehensive metabolic analysis of the major players in GN.

Alterations in metabolic signaling have been implicated in a variety of human diseases. Podocytes, which are part of the kidney filtration barrier, do not represent classical insulin-responsive target cells. However, recent fundamental studies highlighted the pivotal role of insulin signaling in podocytes beyond mere glucose uptake comprising activation of AKT and mTOR.

Figure 2

We have shown that insulin signaling in podocytes is tightly controlled by mitochondrial function. An impairment of the mitochondrial fusion and fission machinery caused by loss of the expression of Oma1 or Phb2 is associated with an increased activation of the insulin signal cascade (3). In a recently published comprehensive analysis podocyte metabolism, we found that anaerobic glycolysis represents the predominant metabolic pathway of podocytes (4).

These findings offer a strategy to therapeutically interfere with the enhanced podocyte metabolism in various progressive kidney diseases, such as diabetic nephropathy or focal segmental glomerulosclerosis (FSGS).

  • Brähler S, Zinselmeyer BH, Raju S, Nitschke M, Suleiman H, Saunders, BT, Johnson MW, Boehner, AMC, Viehmann S, Theisen, DJ, Kretzer MN, Briseño CM, Zaitsev K, Ornatsky O, Chang Q 7, Carrero JA, Kopp JB, Artyomov MN, Kurts C, Murphy KM, Miner JH and Shaw AS.Opposing roles of dendritic cell subsets in experimental glomerulonephritis. J Am Soc Nephrol JASN, 2018, Jan;29(1):138-154.
  • Brähler S*, Yu H*, Suleiman H, Krishnan GM, Saunders BT, Kopp JB, Miner JH, Zinselmeyer BH, Shaw AS.Intravital and kidney slice imaging of podocyte membrane dynamics. J Am Soc Nephrol. JASN 2016 Nov;27(11):3285-3290.
  • Ising C, Koehler S, Brähler S, Merkwirth C, Höhne M, Baris OR, Hagmann H, Kann M, Fabretti F, Dafinger C, Bloch W, Schermer B, Linkermann A, Brüning JC, Kurschat CE, Müller RU, Wiesner RJ, Langer T, Benzing T, BrinkkoetterPT. Inhibition of insulin/IGF-1 receptor signaling protects from mitochondria-mediated kidney failure. EMBO Mol Med. 2015 Feb 2;7(3):275-87
  • Brinkkoetter PT, Bork T, Salou S, Liang W, Mizi A, Özel C, et al. Anaerobic Glycolysis Maintains the Glomerular Filtration Barrier Independent of Mitochondrial Metabolism and Dynamics. Cell Rep. 2019 Apr 30;27(5):1551-1566.e5.
  • Butt L, Unnersjo-Jess D, Hohne M, Hahnfeldt R, Reilly D, Rinschen MM, Plagmann I, Diefenhardt P, Brahler S, Brinkkotter PT, Brismar H, Blom H, Schermer B, and Benzing T (2022). Super-Resolution Imaging of the Filtration Barrier Suggests a Role for Podocin R229Q in Genetic Predisposition to Glomerular Disease. J Am Soc Nephrol33, 138-154. doi:10.1681/ASN.2020060858.
  • Koehler S, Odenthal J, Ludwig V, Unnersjo Jess D, Hohne M, Jungst C, Grawe F, Helmstadter M, Janku JL, Bergmann C, Hoyer PF, Hagmann HH, Walz G, Bloch W, Niessen C, Schermer B, Wodarz A, Denholm B, Benzing T, Iden S, and Brinkkoetter PT (2022). Scaffold polarity proteins Par3A and Par3B share redundant functions while Par3B acts independent of atypical protein kinase C/Par6 in podocytes to maintain the kidney filtration barrier. Kidney Int101, 733-751. doi:10.1016/j.kint.2021.11.030.
  • Kuhne L, Kaufeld J, Volker LA, Wendt R, Schonermarck U, Hagele H, Osterholt T, Eichenauer DA, Bieringer M, von Bergwelt-Baildon A, Fischereder M, Buxhofer-Ausch V, Menne J, Brinkkoetter PT, and Knobl P (2022). Alternate-day dosing of caplacizumab for immune-mediated thrombotic thrombocytopenic purpura. J Thromb Haemost20, 951-960. doi:10.1111/jth.15637.
  • Mukherjee K, Gu C, Collins A, Mettlen M, Samelko B, Altintas MM, Sudhini YR, Wang X, Bouley R, Brown D, Pedro BP, Bane SL, Gupta V, Brinkkoetter PT, Hagmann H, Reiser J, and Sever S (2022). Simultaneous stabilization of actin cytoskeleton in multiple nephron-specific cells protects the kidney from diverse injury. Nat Commun13, 2422. doi:10.1038/s41467-022-30101-4.
  • Eller K, Knoebl P, Bakkaloglu SA, Menne JJ, Brinkkoetter PT, Grandt L, Thiem U, Coppo P, Scully M, and Haller MC (2022). European Renal Best Practice endorsement of guidelines for diagnosis and therapy of thrombotic thrombocytopaenic purpura published by the International Society on Thrombosis and Haemostasis. Nephrol Dial Transplant 37, 1229-1234. doi:10.1093/ndt/gfac034.
  • Kuhne L, Volker LA, Hagmann H, Hagele H, Osterholt T, Eichenauer DA, Thomas A, Breuer J, Gruttner B, Gottschalk I, Kann M, Benzing T, Thevis M, Muller AM, and Brinkkoetter PT (2022). First use of the anti-VWF nanobody caplacizumab to treat iTTP in pregnancy. Br J Haematol 196, e30-e33. doi:10.1111/bjh.17833.
  • Ehren R, Benz MR, Brinkkotter PT, Dotsch J, Eberl WR, Gellermann J, Hoyer PF, Jordans I, Kamrath C, Kemper MJ, Latta K, Muller D, Oh J, Tonshoff B, Weber S, Weber LT, and German Society for Pediatric N (2021). Pediatric idiopathic steroid-sensitive nephrotic syndrome: diagnosis and therapy -short version of the updated German best practice guideline (S2e) - AWMF register no. 166-001, 6/2020. Pediatr Nephrol36, 2971-2985. doi:10.1007/s00467-021-05135-3.
  • Butt L, Unnersjo-Jess D, Hohne M, Hahnfeldt R, Reilly D, Rinschen MM, Plagmann I, Diefenhardt P, Brahler S, Brinkkotter PT, Brismar H, Blom H, Schermer B, and Benzing T (2022). Super-Resolution Imaging of the Filtration Barrier Suggests a Role for Podocin R229Q in Genetic Predisposition to Glomerular Disease. J Am Soc Nephrol33, 138-154. doi:10.1681/ASN.2020060858.
  • Koehler S, Odenthal J, Ludwig V, Jess DU, Hohne M, Jungst C, Grawe F, Helmstadter M, Janku JL, Bergmann C, Hoyer PF, Hagmann HHH, Walz G, Bloch W, Niessen C, Schermer B, Wodarz A, Denholm B, Benzing T, Iden S, and Brinkkoetter PT (2021). Scaffold polarity proteins Par3A and Par3B share redundant functions while Par3B acts independent of atypical protein kinase C/Par6 in podocytes to maintain the kidney filtration barrier. Kidney Int. doi:10.1016/j.kint.2021.11.030.
                                            • Mangold N, Pippin J, Unnersjoe-Jess D, Koehler S, Shankland S, Brahler S, Schermer B, Benzing T, Brinkkoetter PT, and Hagmann H (2021). The Atypical Cyclin-Dependent Kinase 5 (Cdk5) Guards Podocytes from Apoptosis in Glomerular Disease While Being Dispensable for Podocyte Development. Cells10. doi:10.3390/cells10092464.
                                            • Talyan S, Filipow S, Ignarski M, Smieszek M, Chen H, Kuhne L, Butt L, Gobel H, Hoyer-Allo KJR, Koehler FC, Altmuller J, Brinkkotter P, Schermer B, Benzing T, Kann M, Muller RU, and Dieterich C (2021). CALINCA-A Novel Pipeline for the Identification of lncRNAs in Podocyte Disease. Cells10. doi:10.3390/cells10030692.
                                            • Butt L, Unnersjo-Jess D, Hohne M, Edwards A, Binz-Lotter J, Reilly D, Hahnfeldt R, Ziegler V, Fremter K, Rinschen MM, Helmstadter M, Ebert LK, Castrop H, Hackl MJ, Walz G, Brinkkoetter PT, Liebau MC, Tory K, Hoyer PF, Beck BB, Brismar H, Blom H, Schermer B, and Benzing T (2020). A molecular mechanism explaining albuminuria in kidney disease. Nature metabolism 2, 461-74.
                                               
                                            • Kaufeld J, Brinkkoetter PT, Schreiber A, Jabs WJ, Bieringer M, Bruck H, Menne J, and Volker LA (2020). Caplacizumab: frequent local skin reactions. Annals of hematology 10.1007/s00277-020-04260-7.
                                               
                                            • Koehler S, Kuczkowski A, Kuehne L, Jungst C, Hoehne M, Grahammer F, Eddy S, Kretzler M, Beck BB, Hohfeld J, Schermer B, Benzing T, Brinkkoetter PT, and Rinschen MM (2020). Proteome Analysis of Isolated Podocytes Reveals Stress Responses in Glomerular Sclerosis. J Am Soc Nephrol 31, 544-59.
                                               
                                            • Matin M, Morgelin M, Stetefeld J, Schermer B, Brinkkoetter PT, Benzing T, Koch M, and Hagmann H (2020). Affinity-Enhanced Multimeric VEGF (Vascular Endothelial Growth Factor) and PlGF (Placental Growth Factor) Variants for Specific Adsorption of sFlt-1 to Restore Angiogenic Balance in Preeclampsia. Hypertension 76, 1176-84.
                                               
                                            • Volker LA, Brinkkoetter PT, Knobl PN, Krstic M, Kaufeld J, Menne J, Buxhofer-Ausch V, and Miesbach W (2020a). Treatment of acquired thrombotic thrombocytopenic purpura without plasma exchange in selected patients under caplacizumab. Journal of Thrombosis and Haemostasis 18, 3061-6.
                                            • Volker LA, Kaufeld J, Miesbach W, Brahler S, Reinhardt M, Kuhne L, Muhlfeld A, Schreiber A, Gaedeke J, Tolle M, Jabs WJ, Ozcan F, Markau S, Girndt M, Bauer F, Westhoff TH, Felten H, Hausberg M, Brand M, Gerth J, Bieringer M, Bommer M, Zschiedrich S, Schneider J, Elitok S, Gawlik A, Gackler A, Kribben A, Schwenger V, Schoenermarck U, Roeder M, Radermacher J, Bramstedt J, Morgner A, Herbst R, Harth A, Potthoff SA, von Auer C, Wendt R, Christ H, Brinkkoetter PT, and Menne J (2020). ADAMTS13 and VWF activities guide individualized caplacizumab treatment in patients with aTTP. Blood Advances4, 3093-3101. doi:10.1182/bloodadvances.2020001987.
                                            Prof. Dr. Paul T Brinkkötter CMMC Cologne
                                            Prof. Dr. Paul T Brinkkötter

                                            Clinic II of Internal Medicine

                                            CMMC - PI - B 01

                                            Publications - Paul T Brinkkötter

                                            Link to PubMed

                                            Dr. Sebastian Brähler CMMC Cologne
                                            Dr. Sebastian Brähler

                                            Clinic II of Internal Medicine - CMMC Research Building

                                            CMMC - PI - CAP 17

                                            CMMC - Co-PI - B 01

                                            +49 221 478 4429

                                            Clinic II of Internal Medicine - CMMC Research Building

                                            Robert-Koch-Str. 21

                                            50931 Cologne

                                            https://www.kidneyresearchcenter.org/52n31/Inflammatory-cells-and-glomerular-injury.htm

                                            CMMC Profile Page

                                            Curriculum Vitae (CV)

                                            Publications on PubMed

                                            Publications - Sebastian Brähler

                                            Link to PubMed

                                            Group Members

                                            Paul Diefenhardt (PostDoc)        
                                            Henning Hagmann (PostDoc)
                                            Cem Özel (PostDoc)
                                            Jasper Nies (PostDoc
                                            Marie-Kristin Kroll (PhD student)
                                            Johanna Odenthal (PhD student)
                                            Sachiko Kolar (MTA)
                                            Angelika Köser (MTA)
                                            Duc Nguyen Minh (MD student)
                                            Bastian Trinsch (MD student)