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Assessment of increased glomerular permeability associated with recurrent focal segmental glomerulosclerosis using an in vitro model of the glomerular filtration barrier

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Abstract

The presence of circulating permeability factors (cPFs) has been hypothesized to be associated with recurrence of focal segmental glomerulosclerosis (rFSGS) in renal allografts. The available methods to detect cPFs are complex, not easily repeatable and inappropriate to represent the anatomical characteristics of the three-layer glomerular filtration barrier (GFB). Here we describe a novel method which measures the permeability to bovine serum albumin (BSA) through a three-layer device (3LD). The 3 layers comprise: (1) conditionally immortalized human podocytes (HCiPodo), (2) collagen type IV coated porous membrane and (3) human glomerular endothelial cells (HCiGEnC). Using this method, we found that sera from all rFSGS patients increased albumin permeability, while sera from non recurrent (nrFSGS) and genetic (gFSGS) forms of FSGS did not. The mechanisms underlying the increase of albumin permeability are probably due to endothelial cell damage as an initial event, which was demonstrated by the decrease of Platelet endothelial cell adhesion molecule (PECAM-1 or CD31), while the podocytes’ expressions of synaptopodin and podocin were normal. Furthermore, we also found that the plasmapheretic treatment (PPT) eliminated the effect of increasing BSA permeability in sera from rFSGS patients. These preliminary data suggest that our in vitro GFB model could not only be useful in predicting the recurrence of FSGS after renal transplantation (RTx), but also be a valuable in vitro model to study podocyte and endothelial cell biology.

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Acknowledgements

This study was funded by Fondazione IRCCS Ca’ Granda Current Research, Associazione Bambino Nefropatico and Fondazione La Nuova Speranza.

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Authors and Affiliations

  1. IRCCS Ospedale Maggiore Policlinico, Renal Research Laboratory, Foundation Ca’ Granda, Milan, Italy

    Min Li, Silvia Armelloni & Deborah Mattinzoli

  2. Unit of Adult Nephrology, Dialysis and Renal Transplant, Department of Medicine, Foundation Ca’ Granda IRCCS Ospedale Maggiore Policlinico, Via Commenda 15, 20122, Milan, Italy

    Carlo Maria Alfieri & Piergiorgio Messa

  3. Pediatric Nephrology, Dialysis and Transplant Unit, Foundation IRCCS Cà Granda, IRCCS Ospedale Maggiore Policlinico, Milan, Italy

    William Morello & Giovanni Montini

  4. Department of Clinical Sciences and Community Health, Università degli studi di Milano, Milan, Italy

    Carlo Maria Alfieri, Giovanni Montini & Piergiorgio Messa

  5. Politecnico di Milano, Dipartimento di Chimica, Materiali ed Ingegneria Chimica “G. Natta”, Milan, Italy

    Francesco Cellesi

Authors
  1. Min Li
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  2. Carlo Maria Alfieri
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  3. William Morello
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  4. Francesco Cellesi
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  5. Silvia Armelloni
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  6. Deborah Mattinzoli
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  7. Giovanni Montini
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  8. Piergiorgio Messa
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Corresponding author

Correspondence to Piergiorgio Messa.

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Conflict of interest

All the authors declared no competing interests. Dr Li and Dr Rastaldi are the inventors of the 3D co-culture method (Patent application PCT/IB2010/003298, 12/2010, filed by Fondazione IRCCS Ospedale Maggiore Policlinico di Milano; Italian Patent December 28, 2012, No. 0001397084; European Patent August 10, 2016, No. EP2513295).

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Ethical approval for this study was obtained from the ethical committee of Fondazione IRCCS Ca’ Granda Ospedale Maggiore – Policlnico of Milan (Ref: 1004_2019).

Informed consent

Informed consent was obtained from all individual participants included in the study and signed by adult legal guardians in case of minor age patients. The informed consent of involved patients had been previously approved by the EC of Fondazione IRCCS Ca’ Granda Ospedale Maggiore—Policlnico of Milan (ITA) [M. 02. F (A)].

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Li, M., Alfieri, C.M., Morello, W. et al. Assessment of increased glomerular permeability associated with recurrent focal segmental glomerulosclerosis using an in vitro model of the glomerular filtration barrier. J Nephrol 33, 747–755 (2020). https://doi.org/10.1007/s40620-019-00683-2

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  • DOI: https://doi.org/10.1007/s40620-019-00683-2

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