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TNFα pathway blockade ameliorates toxic effects of FSGS plasma on podocyte cytoskeleton and β3 integrin activation

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Abstract

Background

In the absence of mutant genes encoding components of the podocyte slit diaphragm, about 30–50 % of children with primary glucocorticoid-resistant focal segmental glomerulosclerosis (FSGS) develop recurrent proteinuria and slowly progressive FSGS lesions following renal transplantation. Recurrence of FSGS in the allograft strongly suggests a circulating factor that disturbs normal podocyte biology. To date, the nature of the circulating factor is unclear, and there is no cure for the recurrent form of FSGS (R-FSGS).

Methods

Cultured differentiated human podocytes were exposed to the plasmapheresis effluent or blood plasma samples from pediatric patients with recurrent or primary FSGS; in some cases, podocytes were pre-incubated with specific antibodies to block the tumor necrosis factor-alpha (TNFα) signaling pathway. Integrity of focal adhesion complexes and actin cytoskeleton were investigated by immunofluorescent microscopy.

Results

Plasmapheresis effluent from an R-FSGS child or fresh plasma from two children with primary FSGS rapidly disturbed the cytoskeleton of normal human podocytes in vitro. Plasma from a child with R-FSGS also activated β3 integrin and dispersed focal adhesion complexes. The effects were reversed by pre-incubation with antibodies against TNFα or either of the two TNFα receptors. When our patient with R-FSGS became resistant to plasmapheresis, we initiated treatment with twice weekly etanercept injections and then infliximab. Within 3 weeks of regular anti-TNFα therapy, the patient achieved sustained partial remission of proteinuria, allowing us to wean her off plasmapheresis completely.

Conclusions

We suggest that in some FSGS patients, disruption of the podocyte cytoskeleton and β3 integrin-mediated podocyte attachment are driven by the TNFα pathway.

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Acknowledgments

We are indebted to the patients and their families for participating in the study. We thank LeeLee Chu, Annes Siji, and Ashwini Raghavendra for their excellent technical assistance with the Bangalore samples. The study was supported by an infrastructure support grant to the McGill University Health Centre Research Institute from the Fonds de la Recherche du Québec-Santé and by an operating grant to ET from the Canadian Institutes of Health Research. PG is the recipient of a James McGill Research Chair. ET is the recipient of the Fonds de la Recherche du Québec-Santé Chercheur Boursier Junior award. Collaboration between the Montreal Children’s Hospital and St. John’s Medical College Hospital (Bangalore) was supported by an International Society of Nephrology Sister Centre grant.

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Authors and Affiliations

  1. Department of Pediatrics, Montreal Children’s Hospital, McGill University, Montreal, Quebec, Canada

    Martin Bitzan & Paul Goodyer

  2. Department of Medicine, McGill University, Montreal, Quebec, Canada

    Sima Babayeva & Elena Torban

  3. St. John’s Medical College Hospital, Bangalore, India

    Anil Vasudevan

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  1. Martin Bitzan
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  3. Anil Vasudevan
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  4. Paul Goodyer
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  5. Elena Torban
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Corresponding author

Correspondence to Elena Torban.

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Supplementary Figure 1

Blockade of the TNFα pathway restores actin arrangement in cultured podocytes. Differentiated podocytes were incubated for 12 h with 5 % plasma from a healthy age-matched child (upper figure) or 5 % plasma from two children with primary glucocorticoid-resistant FSGS. Pre-incubation of podocytes for 1 h with 0.1 μg/mL of anti-TNFα or 0.2 μg/mL of anti-TNFRI antibodies restored filamentous actin (phalloidin, red) and podocyte shape. (JPG 396 kb)

Supplementary Table 1

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Bitzan, M., Babayeva, S., Vasudevan, A. et al. TNFα pathway blockade ameliorates toxic effects of FSGS plasma on podocyte cytoskeleton and β3 integrin activation. Pediatr Nephrol 27, 2217–2226 (2012). https://doi.org/10.1007/s00467-012-2163-3

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