The role of filamins in mechanically stressed podocytes.
Actin Cytoskeleton
/ metabolism
Adult
Aged
Aged, 80 and over
Animals
Case-Control Studies
Cell Adhesion
Cell Movement
Diabetic Nephropathies
/ metabolism
Filamins
/ metabolism
Focal Adhesions
/ metabolism
Humans
Kidney Glomerulus
/ metabolism
Mice
Middle Aged
Podocytes
/ metabolism
Retrospective Studies
Signal Transduction
Stress, Mechanical
Flna
diabetic nephropathy
mechanical load
mechanosensor
mechanotransducer
Journal
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
ISSN: 1530-6860
Titre abrégé: FASEB J
Pays: United States
ID NLM: 8804484
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
revised:
26
02
2021
received:
19
05
2020
accepted:
15
03
2021
entrez:
16
4
2021
pubmed:
17
4
2021
medline:
28
7
2021
Statut:
ppublish
Résumé
Glomerular hypertension induces mechanical load to podocytes, often resulting in podocyte detachment and the development of glomerulosclerosis. Although it is well known that podocytes are mechanosensitive, the mechanosensors and mechanotransducers are still unknown. Since filamin A, an actin-binding protein, is already described to be a mechanosensor and mechanotransducer, we hypothesized that filamins could be important for the outside-in signaling as well as the actin cytoskeleton of podocytes under mechanical stress. In this study, we demonstrate that filamin A is the main isoform of the filamin family that is expressed in cultured podocytes. Together with filamin B, filamin A was significantly up-regulated during mechanical stretch (3 days, 0.5 Hz, and 5% extension). To study the role of filamin A in cultured podocytes under mechanical stress, filamin A was knocked down (Flna KD) by specific siRNA. Additionally, we established a filamin A knockout podocyte cell line (Flna KO) by CRISPR/Cas9. Knockdown and knockout of filamin A influenced the expression of synaptopodin, a podocyte-specific protein, focal adhesions as well as the morphology of the actin cytoskeleton. Moreover, the cell motility of Flna KO podocytes was significantly increased. Since the knockout of filamin A has had no effect on cell adhesion of podocytes during mechanical stress, we simultaneously knocked down the expression of filamin A and B. Thereby, we observed a significant loss of podocytes during mechanical stress indicating a compensatory mechanism. Analyzing hypertensive mice kidneys as well as biopsies of patients suffering from diabetic nephropathy, we found an up-regulation of filamin A in podocytes in contrast to the control. In summary, filamin A and B mediate matrix-actin cytoskeleton interactions which are essential for the adaptation of cultured podocyte to mechanical stress.
Identifiants
pubmed: 33860543
doi: 10.1096/fj.202001179RR
doi:
Substances chimiques
Filamins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e21560Subventions
Organisme : University Medicine Greifswald
Organisme : University Erlangen
Organisme : University Clinic Hamburg Eppendorf
Organisme : University of Munich
Informations de copyright
© 2021 Federation of American Societies for Experimental Biology.
Références
Global, regional, and national burden of chronic kidney disease, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet (London, England). 2020;395:709-733.
Saran R, Li Y, Robinson B, et al. US renal data system 2014 annual data report: epidemiology of kidney disease in the United States. Am J Kidney Dis. 2015;66, Svii. S1-S305.
Hill NR, Fatoba ST, Oke JL, et al. Global prevalence of chronic kidney disease - a systematic review and meta-analysis. PLoS ONE. 2016;11:e0158765.
Kretzler M, Koeppen-Hagemann I, Kriz W. Podocyte damage is a critical step in the development of glomerulosclerosis in the uninephrectomised-desoxycorticosterone hypertensive rat. Vichows Archiv A Pathol Anat. 1994;425:181-193.
Kriz W, Hosser H, Hahnel B, Gretz N, Provoost A. From segmental glomerulosclerosis to total nephron degeneration and interstitial fibrosis: a histopathological study in rat models and human glomerulopathies. Nephrol Dial Transplant. 1998;132781-2798.
Simons JL, Provoost AP, Anderson S, et al. Pathogenesis of glomerular injury in the fawn-hooded rat: early glomerular capillary hypertension predicts glomerular sclerosis. J Am Soc Nephrol: JASN. 1993;1775-1782. https://jasn.asnjournals.org/content/jnephrol/3/11/1775.full.pdf.
van Dokkum RPE, Sun C-W, Provoost AP, Jacob HJ, Roman RJ. Altered renal hemodynamics and impaired myogenic responses in the fawn-hooded rat. Am J Physiol-Regul, Integrat Comparat Physiol. 1999;276:R855-R863.
Friedrich C, Endlich N, Kriz W, Endlich K. Podocytes are sensitive to fluid shear stress in vitro. Am J Physiol Renal Physiol. 2006;291:F856-F865.
Endlich N, Kress KR, Reiser J, et al. Podocytes respond to mechanical stress in vitro. J Am Soc Nephrol: JASN. 2001;12:413-422.
Gorlin JB, Yamin R, Egan S, et al. Human endothelial actin-binding protein (ABP-280, nonmuscle filamin): a molecular leaf spring. J Cell Biol. 1990;111:1089-1105.
Stossel TP, Condeelis J, Cooley L, et al. Filamins as integrators of cell mechanics and signalling. Nat Rev Mol Cell Biol. 2001;2:138-145.
Nakamura F, Stossel TP, Hartwig JH. The filamins: organizers of cell structure and function. Cell Adh Migr. 2011;5:160-169.
Kim H, McCulloch CA. Filamin A mediates interactions between cytoskeletal proteins that control cell adhesion. FEBS Lett. 2011;585:18-2.
Chakarova C, Wehnert MS, Uhl K, et al. Genomic structure and fine mapping of the two human filamin gene paralogues FLNB and FLNC and comparative analysis of the filamin gene family. Hum Genet. 2000;107:597-611.
Takafuta T, Wu G, Murphy GF, Shapiro SS. Human β-filamin is a new protein that interacts with the cytoplasmic tail of glycoprotein Ibα. J Biol Chem. 1998;273:17531-17538.
Thompson TG, Chan Y-M, Hack AA, et al. Filamin 2 (FLN2): a muscle-specific sarcoglycan interacting protein. J Cell Biol. 2000;148:115-126.
Seppälä J, Tossavainen H, Rodic N, Permi P, Pentikäinen U, Ylänne J. Flexible structure of peptide-bound filamin A mechanosensor domain pair 20-21. PLoS ONE. 2015;10:e0136969.
Rognoni L, Stigler J, Pelz B, Ylänne J, Rief M. Dynamic force sensing of filamin revealed in single-molecule experiments. Proc Natl Acad Sci USA. 2012;109:19679-19684.
Chen H, Chandrasekar S, Sheetz MP, Stossel TP, Nakamura F, Yan J. Mechanical perturbation of filamin A immunoglobulin repeats 20-21 reveals potential non-equilibrium mechanochemical partner binding function. Sci Rep. 2013;3:1642.
Lad Y, Kiema T, Jiang P, et al. Structure of three tandem filamin domains reveals auto-inhibition of ligand binding. EMBO J. 2007;26:3993-4004.
Kiema T, Lad Y, Jiang P, et al. The molecular basis of filamin binding to integrins and competition with talin. Mol Cell. 2006;21:337-347.
Ehrlicher AJ, Nakamura F, Hartwig JH, Weitz DA, Stossel TP. Mechanical strain in actin networks regulates FilGAP and integrin binding to filamin A. Nature. 2011;478:260-263.
Campa CC, Ciraolo E, Ghigo A, Germena G, Hirsch E. Crossroads of PI3K and Rac pathways. Small GTPases. 2015;6:71-80.
Lawson CD, Burridge K. The on-off relationship of Rho and Rac during integrin-mediated adhesion and cell migration. Small GTPases. 2014;5:e27958.
Nakamura F, Heikkinen O, Pentikäinen OT, et al. Molecular basis of filamin A-FilGAP interaction and its impairment in congenital disorders associated with filamin A mutations. PLoS ONE. 2009;4:e4928.
Shifrin Y, Arora PD, Ohta Y, Calderwood DA, McCulloch CA. The role of FilGAP-filamin A interactions in mechanoprotection. Mol Biol Cell. 2009;20:1269-1279.
Kanasaki K, Kanda Y, Palmsten K, et al. Integrin beta1-mediated matrix assembly and signaling are critical for the normal development and function of the kidney glomerulus. Dev Biol. 2008;313:584-593.
Pozzi A, Jarad G, Moeckel GW, et al. Beta1 integrin expression by podocytes is required to maintain glomerular structural integrity. Dev Biol. 2008;316:288-301.
Pozzi A, Zent R. Integrins in kidney disease. J Am Soc Nephrol: JASN. 2013;24:1034-1039.
Liu J, Das M, Yang J, et al. Structural mechanism of integrin inactivation by filamin. Nat Struct Mol Biol. 2015;22:383-389.
MacPherson M, Fagerholm SC. Filamin and filamin-binding proteins in integrin-regulation and adhesion. Focus on: “FilaminA is required for vimentin-mediated cell adhesion and spreading”. Am J Physiol Cell Physiol. 2010;298:C206-C208.
Nakamura F, Osborn TM, Hartemink CA, Hartwig JH, Stossel TP. Structural basis of filamin A functions. J Cell Biol. 2007;179:1011-1025.
Holstein-Rathlou NH, Wagner AJ, Marsh DJ. Tubuloglomerular feedback dynamics and renal blood flow autoregulation in rats. Am J Physiol-Renal Physiol. 1991;260:F53.
Endlich N, Endlich K. The challenge and response of podocytes to glomerular hypertension. Semin Nephrol. 2012;32:327-341.
Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29:e45.
Schindelin J, Arganda-Carreras I, Frise E, et al. Fiji: an open-source platform for biological-image analysis. Nat Methods. 2012;9:676-682.
Rueden CT, Schindelin J, Hiner MC, et al. Image J2: ImageJ for the next generation of scientific image data. BMC Bioinformatics. 2017;18:529.
Cohen CD, Frach K, Schlöndorff D, Kretzler M. Quantitative gene expression analysis in renal biopsies: a novel protocol for a high-throughput multicenter application. Kidney Int. 2002;61:133-140.
Martini S, Nair V, Keller BJ, et al. Integrative biology identifies shared transcriptional networks in CKD. J Am Soc Nephrol: JASN. 2014;25:2559-2572.
Cohen CD, Klingenhoff A, Boucherot A, et al. Comparative promoter analysis allows de novo identification of specialized cell junction-associated proteins. Proc Natl Acad Sci USA. 2006;103:5682-5687.
Tusher VG, Tibshirani R, Chu G. Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA. 2001;98:5116-5121.
Püspöki Z, Storath M, Sage D, Unser M. Transforms and operators for directional bioimage analysis: a survey. Adv Anat Embryol Cell Biol. 2016;219:69-93.
Rezakhaniha R, Agianniotis A, Schrauwen JTC, et al. Experimental investigation of collagen waviness and orientation in the arterial adventitia using confocal laser scanning microscopy. Biomech Model Mechanobiol. 2012;11:461-473.
Fonck E, Feigl GG, Fasel J, et al. Effect of aging on elastin functionality in human cerebral arteries. Stroke. 2009;40:2552-2556.
Kliewe F, Scharf C, Rogge H, et al. Studying the role of fascin-1 in mechanically stressed podocytes. Sci Rep. 2017;7:9916.
Kliewe F, Kaling S, Lötzsch H, et al. Fibronectin is up-regulated in podocytes by mechanical stress. FASEB J. 2019;33:14450-14460.
Hodgin JB, Nair V, Zhang H, et al. Identification of cross-species shared transcriptional networks of diabetic nephropathy in human and mouse glomeruli. Diabetes. 2013;62:299-308.
Lindenmeyer MT, Eichinger F, Sen K, et al. Systematic analysis of a novel human renal glomerulus-enriched gene expression dataset. PLoS One. 2010;5:e11545.
Endlich N, Schordan E, Cohen C, et al. Palladin is a dynamic actin-associated protein in podocytes. Kidney Int. 2009;75:214-226.
Endlich K, Kliewe F, Endlich N. Stressed podocytes-mechanical forces, sensors, signaling and response. Pflugers Arch. 2017;469:937-949.
D'Addario M, Arora PD, Fan J, Ganss B, Ellen RP, McCulloch CA. Cytoprotection against mechanical forces delivered through beta 1 integrins requires induction of filamin A. J Biol Chem. 2001;276:31969-31977.
He F-F, Chen S, Su H, Meng X-F, Zhang C. Actin-associated proteins in the pathogenesis of podocyte injury. Curr Genomics. 2013;14:477-484.
Mathieson PW. The podocyte cytoskeleton in health and in disease. Clin Kidney J. 2012;5:498-501.
Ichimura K, Kurihara H, Sakai T. Actin filament organization of foot processes in rat podocytes. J Histochem Cytochem. 2003;51:1589-1600.
Tseng Y, An KM, Esue O, Wirtz D. The bimodal role of filamin in controlling the architecture and mechanics of F-actin networks. J Biol Chem. 2004;279:1819-1826.
Wang K, Singer SJ. Interaction of filamin with f-actin in solution. Proc Natl Acad Sci USA. 1977;74:2021-2025.
Kumar A, Shutova MS, Tanaka K, et al. Filamin A mediates isotropic distribution of applied force across the actin network. J Cell Biol. 2019;218:2481-2491.
Asanuma K, Yanagida-Asanuma E, Faul C, Tomino Y, Kim K, Mundel P. Synaptopodin orchestrates actin organization and cell motility via regulation of RhoA signalling. Nat Cell Biol. 2006;8:485-491.
Wang L, Nakamura F. Identification of Filamin A mechanobinding partner I: smoothelin specifically interacts with the filamin A mechanosensitive domain 21. Biochemistry. 2019;58:4726-4736.
Mundel P, Heid HW, Mundel TM, Krüger M, Reiser J, Kriz W. Synaptopodin: an actin-associated protein in telencephalic dendrites and renal podocytes. J Cell Biol. 1997;139:193-204.
Hugo C, Nangaku M, Shankland SJ, et al. The plasma membrane-actin linking protein, ezrin, is a glomerular epithelial cell marker in glomerulogenesis, in the adult kidney and in glomerular injury. Kidney Int. 1998;54:1934-1944.
Artelt N, Ludwig TA, Rogge H, et al. The role of palladin in podocytes. J Am Soc Nephrol: JASN. 2018;29:1662-1678.
Brown EJ, Schlöndorff JS, Becker DJ, et al. Mutations in the formin gene INF2 cause focal segmental glomerulosclerosis. Nat Genet. 2010;42:72-76.
Pinto VI, Senini VW, Wang Y, Kazembe MP, McCulloch CA. Filamin A protects cells against force-induced apoptosis by stabilizing talin- and vinculin-containing cell adhesions. FASEB J. 2014;28:453-463.
Mezawa M, Pinto VI, Kazembe MP, Lee WS, McCulloch CA. Filamin A regulates the organization and remodeling of the pericellular collagen matrix. FASEB J. 2016;30:3613-3627.
Hu J, Lu J, Goyal A, et al. Opposing FlnA and FlnB interactions regulate RhoA activation in guiding dynamic actin stress fiber formation and cell spreading. Hum Mol Genet. 2017;26:1294-1304.
Sheen VL, Feng Y, Graham D, Takafuta T, Shapiro SS, Walsh CA. Filamin A and Filamin B are co-expressed within neurons during periods of neuronal migration and can physically interact. Hum Mol Genet. 2002;11:2845-2854.
Fox JW, Lamperti ED, Ekşioğlu YZ, et al. Mutations in filamin 1 prevent migration of cerebral cortical neurons in human periventricular heterotopia. Neuron. 1998;21:1315-1325.
Savoy RM, Ghosh PM. The dual role of filamin A in cancer: can't live with (too much of) it, can't live without it. Endocr Relat Cancer. 2013;20:R341-R356.
Shao Q-Q, Zhang T-P, Zhao W-J, et al. Filamin A: insights into its exact role in cancers. Pathol Oncol Res: POR. 2016;22:245-252.
Baldassarre M, Razinia Z, Burande CF, Lamsoul I, Lutz PG, Calderwood DA. Filamins regulate cell spreading and initiation of cell migration. PLoS ONE. 2009;4:e7830.
Xu Y, Bismar TA, Su J, et al. Filamin A regulates focal adhesion disassembly and suppresses breast cancer cell migration and invasion. J Exp Med. 2010;207:2421-2437.
Schordan S, Grisk O, Schordan E, et al. OPN deficiency results in severe glomerulosclerosis in uninephrectomized mice. Am J Physiol Renal Physiol. 2013;304:F1458-F1470.
Wang B, Koh P, Winbanks C, et al. miR-200a prevents renal fibrogenesis through repression of TGF-β2 expression. Diabetes. 2011;60:280-287.
Albrecht T, Schilperoort M, Zhang S, et al. Carnosine attenuates the development of both type 2 diabetes and diabetic nephropathy in BTBR ob/ob mice. Sci Rep. 2017;7:44492.