Calpain Activation and Secretion Promote Glomerular Injury in Experimental Glomerulonephritis

Peltier, Julie; Bellocq, A.; Perez, Joëlle; Doublier, Sophie; Dubois, Yi-Chun Xu; Haymann, Jean‐Philippe; Camussi, Giovanni; Baud, Laurent

doi:10.1681/asn.2006050542

Cited by 56 publications

(76 citation statements)

References 27 publications

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“…Alterations in these signaling pathways, frequently caused by genetic mutations within these networks that are responsible for biochemical dysfunctions in proteins and in their related signaling cascades, may progress to glomerular diseases (Wiggins, 2007;Mathieson, 2008). One of the key components of the podocyte signaling hub is nephrin, which appears to be controlled at multiple levels by the modulation of its phosphorylation state via Src family tyrosine kinases (Fyn, Yes, Src) and trafficking (Quack et al, 2006) or by a direct degradation process mediated by calpain (Peltier et al, 2006;Tian et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Rostafuroxin Protects from Podocyte Injury and Proteinuria Induced by Adducin Genetic Variants and Ouabain

Ferrandi

Molinari

Rastaldi

et al. 2014

J Pharmacol Exp Ther

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Glomerulopathies are important causes of morbidity and mortality. Selective therapies that address the underlying mechanisms are still lacking. Recently, two mechanisms, mutant b-adducin and ouabain, have been found to be involved in glomerular podocytopathies and proteinuria through nephrin downregulation. The main purpose of the present study was to investigate whether rostafuroxin, a novel antihypertensive agent developed as a selective inhibitor of Src-SH2 interaction with mutant adducinand ouabain-activated Na,K-ATPase, may protect podocytes from adducin-and ouabain-induced effects, thus representing a novel pharmacologic approach for the therapy of podocytopathies and proteinuria caused by the aforementioned mechanisms. To study the effect of rostafuroxin on podocyte protein changes and proteinuria, mice carrying mutant b-adducin and ouabain hypertensive rats were orally treated with 100 mg/kg per day rostafuroxin. Primary podocytes from congenic rats carrying mutant a-adducin or b-adducin (NB) from Milan hypertensive rats and normal rat podocytes incubated with 10 29 M ouabain were cultured with 10 29 M rostafuroxin. The results indicated that mutant b-adducin and ouabain caused podocyte nephrin loss and proteinuria in animal models. These alterations were reproduced in primary podocytes from NB rats and normal rats incubated with ouabain. Treatment of animals, or incubation of cultured podocytes with rostafuroxin, reverted mutant b-adducin-and ouabain-induced effects on nephrin protein expression and proteinuria. We conclude that rostafuroxin prevented podocyte lesions and proteinuria due to mutant b-adducin and ouabain in animal models. This suggests a potential therapeutic effect of rostafuroxin in patients with glomerular disease progression associated with these two mechanisms.

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Section: Discussionmentioning

confidence: 99%

Rostafuroxin Protects from Podocyte Injury and Proteinuria Induced by Adducin Genetic Variants and Ouabain

Ferrandi

Molinari

Rastaldi

et al. 2014

J Pharmacol Exp Ther

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“…To examine the role of calpain, we used a fully allogenic skin allograft model and transgenic mice expressing high levels of calpastatin (CalpTG) recently generated in our laboratory, as there is no pharmacological tool yet allowing us to specifically suppress the activity of calpains [12,13]. Our results demonstrate that calpain inhibition in transgenic mice attenuates skin allograft rejection.…”

Section: Introductionmentioning

confidence: 90%

“…by both measuring the calpain-specific cleavage of fluorescent AMC substrate and by measuring the accumulation of 145/150-kDa spectrin BDP by Western blot analysis, as previously described [12,13].…”

Section: Calpain Activity Assaymentioning

confidence: 99%

“…All procedures involving these animals were conducted in accordance with national guidelines and institutional policies. CalpTG mice were created in the laboratory using the cDNA clone of rabbit calpastatin inserted on the PCI expression vector, which includes a viral promoter (CMV immediate-early enhancer/ promoter region) [12,13]. The presence and the expression of the transgene were identified in founder CalpTG mice by PCR and RT-PCR analysis, respectively [12].…”

Section: Mice and Skin Transplantationmentioning

confidence: 99%

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Critical role of the calpain/calpastatin balance in acute allograft rejection

et al. 2010

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Rejection of solid organ allograft involves alloreactive T-cell expansion. The importance of NF-jB and NFAT in this process is underscored by the therapeutic efficacy of immunosuppressive agents, which target the two transcription factors. Since calpains, calciumactivated proteases, are involved in the activation of NF-jB and NFAT, we investigated the role of calpains in allograft rejection. In human transplant kidneys undergoing acute or chronic rejection, we show an increased expression of CAPN 1 gene encoding l-calpain, associated with a marked expression of l-calpain, mainly in infiltrating T cells. To address the role of calpain in rejection, we used a skin transplant model in transgenic mice expressing high levels of calpastatin, a calpain-specific inhibitor. We show that calpain inhibition extended skin allograft survival, from 11 to 20 days. This delay was associated with a limitation in allograft infiltration by T cells. In vitro, calpain inhibition by calpastatin transgene expression limited dramatically T-cell migration but, unexpectedly, increased slightly T-cell proliferation. Amplification of IL-2 signaling via the stabilization of IL-2R common c-chain provided an explanation for the proliferation response. This is the first study establishing that calpain inhibition delays allograft rejection by slowing down T-cell migration rather than proliferation.Key words: Allograft . Calpain . Calpastatin . T cells IntroductionSolid organ transplantation represents an important means of treating end stage organ failure. However, this strategy is limited by the immune response mounted by the recipient against donor tissue. Acute allograft rejection involves T cells of the adaptive immune system in addition to cells of the innate immune system [1]. T-cell receptor (TCR) engagement in naïve T cells initiates changes in gene expression that are essential for the generation of effector T cells. They require the activation of transcription factors, primarily NF-kB and NFAT [2,3]. TCR/CD28-induced NF-kB activation characteristically elicits the expression of both IL-2 and IL-2 receptor a chain together with the antiapoptotic molecule Bcl-x L [2]. NFAT, which is activated by the calmodulin-dependent phosphatase calcineurin, is also required for the expression of the IL-2 gene through its interaction with proteins of the AP1 family of transcription factors [3]. Given the importance of these two pathways in the generation Ã These authors contributed equally to this work. Eur. J. Immunol. 2011. 41: 473-484 DOI 10.1002 Leukocyte signaling 473 of effector T cells, a number of different pharmacological inhibitors of NF-kB and/or calcineurin/NFAT are currently used as immunosuppressive agents in transplantation, including steroids, cyclosporin A and tacrolimus (FK-506) [4]. Calpains are calcium-activated neutral cysteine proteases [5]. Two major isoforms, calpain m (or I) and calpain m (or II), which require micromolar and millimolar Ca 21 concentrations for activity, respectively, are ubiquitously expressed, whereas the ...

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“…Mice-Transgenic C57BL/6 mice constitutively overexpressing rabbit calpastatin (under the control of the cytomegalovirus immediate early enhancer/promoter region) have been described previously (31). Samples of back skin were obtained from 8-week-old transgenic and wild type mice.…”

Section: Nh 2 -Terminal Amino Acid Sequencing-recombinant Flg2mentioning

confidence: 99%

Deimination of Human Filaggrin-2 Promotes Its Proteolysis by Calpain 1

Hsu

Henry

Raymond

et al. 2011

Journal of Biological Chemistry

113

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Filaggrin-2 (FLG2), a member of the S100-fused type protein family, shares numerous features with filaggrin (FLG), a key protein implicated in the epidermal barrier functions. Both display a related structural organization, an identical pattern of expression and localization in human epidermis, and proteolytic processing of a large precursor. Here, we tested whether FLG2 was a substrate of calpain 1, a calcium-dependent protease directly involved in FLG catabolism. In addition, deimination being critical for FLG degradation, we analyzed whether FLG2 deimination interfered with its proteolytic processing. With this aim, we first produced a recombinant form of FLG2 corresponding to subunits B7 to B10 fused to a COOH-terminal His tag. Incubation with calpain 1 in the presence of calcium induced a rapid degradation of the recombinant protein and the production of several peptides, as shown by Coomassie Blue-stained gels and Western blotting with anti-FLG2 or anti-His antibodies. MALDI-TOF mass spectrometry confirmed this result and further evidenced the production of non-immunoreactive smaller peptides. The degradation was not observed when a calpain 1-specific inhibitor was added. The calpain cleavage sites identified by Edman degradation were regularly present in the B-type repeats of FLG2. Moreover, immunohistochemical analysis of normal human skin revealed colocalization of FLG2 and calpain 1 in the upper epidermis. Finally, the FLG2 deiminated by human peptidylarginine deiminases was shown to be more susceptible to calpain 1 than the unmodified protein. Altogether, these data demonstrate that calpain 1 is essential for the proteolytic processing of FLG2 and that deimination accelerates this process.In the epidermis, the program of keratinocyte terminal differentiation is an oriented process during which cells of the basal layer undergo a series of metabolic and structural changes throughout their migration to the surface of the tissue. The stratum corneum, the outermost layer of the epidermis, is formed by the stacking of so-called corneocytes, the end products of the process. The stratum corneum functions as an effective barrier between the body and its outside environment, limiting skin dehydration and preventing the penetration of outside pathogens, UV radiation, and exogenous chemicals. It also contributes to mechanical protection of the body. So that this function can be achieved, peculiar structures are formed, such as the cornified cell envelope, a resistant and insoluble protein shell that replaces the plasma membrane, and the intracorneocyte fibrous matrix made by the aggregation of keratin intermediate filaments.

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Calpain Activation and Secretion Promote Glomerular Injury in Experimental Glomerulonephritis

Cited by 56 publications

References 27 publications

Rostafuroxin Protects from Podocyte Injury and Proteinuria Induced by Adducin Genetic Variants and Ouabain

Rostafuroxin Protects from Podocyte Injury and Proteinuria Induced by Adducin Genetic Variants and Ouabain

Critical role of the calpain/calpastatin balance in acute allograft rejection

Deimination of Human Filaggrin-2 Promotes Its Proteolysis by Calpain 1

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