One Year of Enzyme Replacement Therapy Reduces Globotriaosylceramide Inclusions in Podocytes in Male Adult Patients with Fabry Disease - Fig 5

<p><b>(A)</b> Foot process effacement(black arrows) in a biopsy from a Fabry patient at baseline (ERT-naïve). Asterisk marks a GL-3 inclusion in a podocyte (PC). White arrow shows GL-3 inclusions in an endothelial cell (En). <b>(B)</b> A biopsy after 11–12 months of ERT from the same patient still shows areas of glomerular basement membranes with foot process effacement (black arrows). Note that podocytes contain GL-3 inclusions (asterisk), while the endothelial cells are cleared from inclusions. <b>(C)</b> Intact foot processes (black arrows) from a normal control biopsy. <b>(D)</b> Foot process width (FPW) changes estimated by unbiased morphometry. Although FPW was numerically reduced from baseline to follow up in 4/6 cases, the difference was not statistically significant (p = 0.29). The dashed area shows the range of FPW in biopsies from 9 healthy kidney donor normal controls. FPW in baseline and follow up biopsies from Fabry patients were significantly greater than normal controls (p = 0.002 and p = 0.0006, respectively). <b>(E)</b> Foot process effacement (FPE) semi-quantitative scores did not change significantly from baseline to follow up. FPE scoring was based on Tøndel et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152812#pone.0152812.ref016" target="_blank">16</a>], where a score of “0” = no foot identified process effacement; “(+)” = foot process effacement in short segments; and “+” = foot process effacement in longer segments.</p

Risk of death due to categories of renal disease according to whether or not a first degree relative has end stage renal disease.

<p>Risk of death due to categories of renal disease according to whether or not a first degree relative has end stage renal disease.</p

Various stages of GL3 inclusion exocytosis in podocytes.

<p><b>(A)</b> Fusion of GL3 inclusions or their surrounding membranes with the cell membrane (arrows); <b>(B)</b> The space surrounding an intracellular GL3 inclusion is connected to the urinary space through an orifice (arrow); <b>(C)</b> A portion of a GL3 inclusion is squeezed into the urinary space through an orifice (arrow). An adjacent GL3 inclusion is partially extruded from the podocyte while preserving its round shape (arrowhead): <b>(D)</b> A GL3 inclusions extruding from a podocyte shows partial unfolding of its multilamellar structure (arrow); <b>(E)</b> Unfolding of the multilamellar structure of a GL3 inclusion while being squeezed out of a podocyte through an orifice (arrow); <b>(F)</b> Extruded GL3 inclusions in the urinary space (arrow); <b>(G)</b> A large round GL3 inclusion almost completed its extrusion from the podocyte (arrow); <b>(H)</b> An empty round space in a podocyte with partial protrusion into the urinary space, reflecting recent exocytosis (asterisk) and a nearby extracellular GL3 inclusion (arrow); <b>(I)</b> A large and partially unfolded GL3 inclusion in the urinary space (arrow).</p

Risk of death due to categories of cardiovascular disease, according to whether or not a first degree relative had ESRD.

<p>Risk of death due to categories of cardiovascular disease, according to whether or not a first degree relative had ESRD.</p

Characteristics of the population.

<p>Characteristics of the population.</p

One Year of Enzyme Replacement Therapy Reduces Globotriaosylceramide Inclusions in Podocytes in Male Adult Patients with Fabry Disease - Fig 3

<p><b>(A)</b> A representative glomerulus from an 18 year old male (Case #1, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152812#pone.0152812.t001" target="_blank">Table 1</a> for patient's characteristics) with Fabry disease at baseline (ERT-naïve). Podocytes (PC) are remarkably enlarged with abundant GL3 inclusions (Inc); <b>(B)</b> A representative glomerulus from the same patient after 12 months of ERT (1 mg/kg/EOH agalsidase-beta) shows smaller podocytes. The majority of podocytes still showed many GL3 inclusions (asterisks). However, occasional podocytes showed no GL3 inclusions (Ø); <b>(C)</b>Total volume of GL3 inclusions per podocytes V(Inc/PC) was reduced after 11–12 months of ERT (p = 0.02); <b>(D)</b> Podocyte GL3 score (semiquantitative [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152812#pone.0152812.ref014" target="_blank">14</a>]) did not reduce significantly after 11–12 months (follow-up) of ERT (p = 0.18); <b>(E)</b> Podocyte volume significantly decreased after 11–12 months of ERT compared to baseline (p = 0.02). The dashed area shows the range of podocyte volume in biopsies from 5 healthy kidney donor normal controls. The difference between podocyte volume in Fabry patients after 11–12 months of ERT and these healthy controls was not statistically significant; <b>(F)</b> Podocyte GL3 inclusion volume fraction [Vv(Inc/PC)] did not change significantly after 11–12 months of ERT (p = 0.42). The numbers written by each line in C-F represents case numbers according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152812#pone.0152812.t001" target="_blank">Table 1</a>. Panels C-F show the average values of presented parameters in each biopsy.</p

Risk of death due to selected causes according to whether or not a first degree relative had ESRD.

<p>Risk of death due to selected causes according to whether or not a first degree relative had ESRD.</p

Adjusted Kaplan Meier failure curve for all cause death in individuals with and without a first degree relative with ESRD.

<p>Adjusted Kaplan Meier failure curve for all cause death in individuals with and without a first degree relative with ESRD.</p

Various stages of GL3 inclusion exocytosis in podocytes.

<p><b>(A)</b> Fusion of GL3 inclusions or their surrounding membranes with the cell membrane (arrows); <b>(B)</b> The space surrounding an intracellular GL3 inclusion is connected to the urinary space through an orifice (arrow); <b>(C)</b> A portion of a GL3 inclusion is squeezed into the urinary space through an orifice (arrow). An adjacent GL3 inclusion is partially extruded from the podocyte while preserving its round shape (arrowhead): <b>(D)</b> A GL3 inclusions extruding from a podocyte shows partial unfolding of its multilamellar structure (arrow); <b>(E)</b> Unfolding of the multilamellar structure of a GL3 inclusion while being squeezed out of a podocyte through an orifice (arrow); <b>(F)</b> Extruded GL3 inclusions in the urinary space (arrow); <b>(G)</b> A large round GL3 inclusion almost completed its extrusion from the podocyte (arrow); <b>(H)</b> An empty round space in a podocyte with partial protrusion into the urinary space, reflecting recent exocytosis (asterisk) and a nearby extracellular GL3 inclusion (arrow); <b>(I)</b> A large and partially unfolded GL3 inclusion in the urinary space (arrow).</p

Correlation between volume fraction of GL-3 inclusions per podocyte [Vv(Inc/PC)] % change from baseline to follow up (11–12 months post ERT) and foot process width (FPW) at follow up.

<p>Correlation between volume fraction of GL-3 inclusions per podocyte [Vv(Inc/PC)] % change from baseline to follow up (11–12 months post ERT) and foot process width (FPW) at follow up.</p