TY - JOUR
T1 - Differential expression of inflammasome-related genes in induced pluripotent stem-cell-derived retinal pigment epithelial cells with or without history of age-related macular degeneration
AU - Hytti, Maria
AU - Korhonen, Eveliina
AU - Hongisto, Heidi
AU - Kaarniranta, Kai
AU - Skottman, Heli
AU - Kauppinen, Anu
N1 - Funding Information:
Funding: This research was funded by the Academy of Finland (HS and HH (315085), KK (296840), AK (297267, 328443)), the Emil Aaltonen Foundation, the Sigrid Juselius Foundation, the Mary and Georg C. Ehrnrooth Foundation, the Kuopio University Hospital (Grant Number 5503743), the Finnish Eye Foundation, and the Päivikki and Sakari Sohlberg Foundation.
Funding Information:
This research was funded by the Academy of Finland (HS and HH (315085), KK (296840), AK (297267, 328443)), the Emil Aaltonen Foundation, the Sigrid Juselius Foundation, the Mary and Georg C. Ehrnrooth Foundation, the Kuopio University Hospital (Grant Number 5503743), the Finnish Eye Foundation, and the P?ivikki and Sakari Sohlberg Foundation. Laboratory technicians Outi Melin and Hanna Pekkanen, Tampere University, are thanked for excellent assistance with iPSC and iPSC-RPE cell culture and analyses. The authors wish to thank laboratory technician Anne Sepp?nen, University of Eastern Finland, for excellent technical assistance. The authors acknowledge the Tampere Imaging Facility (TIF), Tampere University, Finland for their services. Ewen MacDonald is acknowledged for his assistance in correcting the language of the manuscript.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/6
Y1 - 2021/6
N2 - Inflammation is a key underlying factor of age-related macular degeneration (AMD) and inflammasome activation has been linked to disease development. Induced pluripotent stem-cell-derived retinal pigment epithelial cells (iPSC-RPE) are an attractive novel model system that can help to further elucidate disease pathways of this complex disease. Here, we analyzed the effect of dysfunctional protein clearance on inflammation and inflammasome activation in iPSC-RPE cells generated from a patient suffering from age-related macular degeneration (AMD) and an age-matched control. We primed iPSC-RPE cells with IL-1α and then inhibited both proteasomal degradation and autophagic clearance using MG-132 and bafilomycin A1, respectively, causing inflammasome activation. Subsequently, we determined cell viability, analyzed the expression levels of inflammasome-related genes using a PCR array, and measured the levels of pro-inflammatory cytokines IL-1β, IL-6, IL-8, and MCP-1 secreted into the medium. Cell treatments modified the expression of 48 inflammasome-related genes and increased the secretion of mature IL-1β, while reducing the levels of IL-6 and MCP-1. Interestingly, iPSC-RPE from an AMD donor secreted more IL-1β and expressed more Hsp90 prior to the inhibition of protein clearance, while MCP-1 and IL-6 were reduced at both protein and mRNA levels. Overall, our results suggest that cellular clearance mechanisms might already be dysfunctional, and the inflammasome activated, in cells with a disease origin.
AB - Inflammation is a key underlying factor of age-related macular degeneration (AMD) and inflammasome activation has been linked to disease development. Induced pluripotent stem-cell-derived retinal pigment epithelial cells (iPSC-RPE) are an attractive novel model system that can help to further elucidate disease pathways of this complex disease. Here, we analyzed the effect of dysfunctional protein clearance on inflammation and inflammasome activation in iPSC-RPE cells generated from a patient suffering from age-related macular degeneration (AMD) and an age-matched control. We primed iPSC-RPE cells with IL-1α and then inhibited both proteasomal degradation and autophagic clearance using MG-132 and bafilomycin A1, respectively, causing inflammasome activation. Subsequently, we determined cell viability, analyzed the expression levels of inflammasome-related genes using a PCR array, and measured the levels of pro-inflammatory cytokines IL-1β, IL-6, IL-8, and MCP-1 secreted into the medium. Cell treatments modified the expression of 48 inflammasome-related genes and increased the secretion of mature IL-1β, while reducing the levels of IL-6 and MCP-1. Interestingly, iPSC-RPE from an AMD donor secreted more IL-1β and expressed more Hsp90 prior to the inhibition of protein clearance, while MCP-1 and IL-6 were reduced at both protein and mRNA levels. Overall, our results suggest that cellular clearance mechanisms might already be dysfunctional, and the inflammasome activated, in cells with a disease origin.
KW - Age-related macular degeneration
KW - Induced pluripotent stem cells
KW - Inflammasomes
KW - Inflammation
KW - Retinal pigment epithelium
U2 - 10.3390/ijms22136800
DO - 10.3390/ijms22136800
M3 - Article
AN - SCOPUS:85108401869
SN - 1661-6596
VL - 22
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 13
M1 - 6800
ER -