TY - JOUR
T1 - Evaluation of the efficacy of cystinosin supplementation through CTNS mRNA delivery in experimental models for cystinosis
AU - Bondue, Tjessa
AU - Berlingerio, Sante Princiero
AU - Siegerist, Florian
AU - Sendino-Garví, Elena
AU - Schindler, Maximilian
AU - Baelde, Hans Jacobus
AU - Cairoli, Sara
AU - Goffredo, Bianca Maria
AU - Arcolino, Fanny Oliveira
AU - Dieker, J. rgen
AU - Janssen, Manoe Jacoba
AU - Endlich, Nicole
AU - Brock, Roland
AU - Gijsbers, Rik
AU - van den Heuvel, Lambertus
AU - Levtchenko, Elena
N1 - Funding Information: The authors thank Marianne Klawitter (Department of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Germany), Laleh Khodaparast and Ladan Khodaparast (Switch Laboratory, KU Leuven, Belgium and VIB Center for Brain & Disease Research, Leuven, Belgium), Martin Lowe (Faculty of Biology, University of Manchester, The United Kingdom), Peter de Witte and Jan Maes (Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Belgium), Kathleen Lambaerts, Frédéric Hendrickx and Kim van Kelst (Aquatic Facility, KU Leuven, Belgium) for their assistance and contribution in the zebrafish experiments. The authors gratefully acknowledge the VIB Bio Imaging Core (VIB Leuven, Belgium) for their support and assistance in this work and Pieter Vanden Berghe (Confocal Imaging Cluster (CIC), KU Leuven, Belgium) for the usage of the confocal microscope (supported by Hercules AKUL/15/37_GOH1816N and FWO G.0929.15 to Pieter Vanden Berghe, KU Leuven). Funding Information: The authors thank Marianne Klawitter (Department of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Germany), Laleh Khodaparast and Ladan Khodaparast (Switch Laboratory, KU Leuven, Belgium and VIB Center for Brain & Disease Research, Leuven, Belgium), Martin Lowe (Faculty of Biology, University of Manchester, The United Kingdom), Peter de Witte and Jan Maes (Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Belgium), Kathleen Lambaerts, Frédéric Hendrickx and Kim van Kelst (Aquatic Facility, KU Leuven, Belgium) for their assistance and contribution in the zebrafish experiments. The authors gratefully acknowledge the VIB Bio Imaging Core (VIB Leuven, Belgium) for their support and assistance in this work and Pieter Vanden Berghe (Confocal Imaging Cluster (CIC), KU Leuven, Belgium) for the usage of the confocal microscope (supported by Hercules AKUL/15/37_GOH1816N and FWO G.0929.15 to Pieter Vanden Berghe, KU Leuven). Funding Information: The authors express gratitude to F.W.O Vlaanderen, for the support to Tjessa Bondue (11A7821N and 11A7823N) and Elena Levtchenko (1801120N). We acknowledge the KU Leuven C1 Grant to Elena Levtchenko, Lambertus van den Heuvel and Rik Gijsbers (C14/17/11) and the Federal Ministry of Education and Research (BMBF) and STOP-FSGS Research Network for their support to Nicole Endlich (01GM1518B). We acknowledge Cystinosis Ireland and the Cystinosis Foundation UK co-funded Research Award 2021. The collaboration project is co-funded by the PPP Allowance made available by Health ~ Holland (Geval 2020–2022), Top Sector Life Sciences & Health, to stimulate public–private partnerships. Elena Levtchenko is a member of the European Reference Kidney Network (ERKNet). Publisher Copyright: © 2023, The Author(s).
PY - 2023/12/1
Y1 - 2023/12/1
N2 - Messenger RNA (mRNA) therapies are emerging in different disease areas, but have not yet reached the kidney field. Our aim was to study the feasibility to treat the genetic defect in cystinosis using synthetic mRNA in cell models and ctns −/− zebrafish embryos. Cystinosis is a prototype lysosomal storage disorder caused by mutations in the CTNS gene, encoding the lysosomal cystine-H+ symporter cystinosin, and leading to cystine accumulation in all cells of the body. The kidneys are the first and the most severely affected organs, presenting glomerular and proximal tubular dysfunction, progressing to end-stage kidney failure. The current therapeutic standard cysteamine, reduces cystine levels, but has many side effects and does not restore kidney function. Here, we show that synthetic mRNA can restore lysosomal cystinosin expression following lipofection into CTNS −/− kidney cells and injection into ctns −/− zebrafish. A single CTNS mRNA administration decreases cellular cystine accumulation for up to 14 days in vitro. In the ctns −/− zebrafish, CTNS mRNA therapy improves proximal tubular reabsorption, reduces proteinuria, and restores brush border expression of the multi-ligand receptor megalin. Therefore, this proof-of-principle study takes the first steps in establishing an mRNA-based therapy to restore cystinosin expression, resulting in cystine reduction in vitro and in the ctns −/− larvae, and restoration of the zebrafish pronephros function.
AB - Messenger RNA (mRNA) therapies are emerging in different disease areas, but have not yet reached the kidney field. Our aim was to study the feasibility to treat the genetic defect in cystinosis using synthetic mRNA in cell models and ctns −/− zebrafish embryos. Cystinosis is a prototype lysosomal storage disorder caused by mutations in the CTNS gene, encoding the lysosomal cystine-H+ symporter cystinosin, and leading to cystine accumulation in all cells of the body. The kidneys are the first and the most severely affected organs, presenting glomerular and proximal tubular dysfunction, progressing to end-stage kidney failure. The current therapeutic standard cysteamine, reduces cystine levels, but has many side effects and does not restore kidney function. Here, we show that synthetic mRNA can restore lysosomal cystinosin expression following lipofection into CTNS −/− kidney cells and injection into ctns −/− zebrafish. A single CTNS mRNA administration decreases cellular cystine accumulation for up to 14 days in vitro. In the ctns −/− zebrafish, CTNS mRNA therapy improves proximal tubular reabsorption, reduces proteinuria, and restores brush border expression of the multi-ligand receptor megalin. Therefore, this proof-of-principle study takes the first steps in establishing an mRNA-based therapy to restore cystinosin expression, resulting in cystine reduction in vitro and in the ctns −/− larvae, and restoration of the zebrafish pronephros function.
UR - http://www.scopus.com/inward/record.url?scp=85177775144&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/s41598-023-47085-w
DO - https://doi.org/10.1038/s41598-023-47085-w
M3 - Article
C2 - 38016974
SN - 2045-2322
VL - 13
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 20961
ER -