TY - JOUR
T1 - Modeling Choroideremia Disease with Isogenic Induced Pluripotent Stem Cells
AU - Fonseca, Ana Fragoso
AU - Coelho, Rita
AU - Lopes-Da-Silva, Mafalda
AU - Lemos, Luísa
AU - Hall, Michael J.
AU - Oliveira, Daniela
AU - Falcão, Ana Sofia
AU - Tenreiro, Sandra
AU - Seabra, Miguel C.
AU - Antas, Pedro
N1 - Funding Information:
The authors declare financial support was received for the research, authorship, and/or publication of this article. Research was supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (FCT)\u2014Portugal (including iNOVA4Health projects UIDB/04462/2020, UIDP/04462/2020, and associated laboratory LS4FUTURE with reference LA/P/0087/2020). A.F.F. is funded by FCT PhD studentship (2022.12254.BD). M.L.S. is funded by the individual grant CEECIND/01536/2018. A.S.F. postdoctoral contract is funded by \u201CLa Caixa Foundation\u201D (NASCENT HR22-00569). P.A. is supported by grant EXPL/MED-OUT/0599/2021, funded by national funds from FCT/MCTES, and individual grant CEECIND/03862/2020. L.L. is a recipient of a Choroideremia Research Foundation postdoctoral fellowship, Throssell-Hillier Research Award. Research at the Center for the Unknown is supported by Funda\u00E7\u00E3o Champalimaud.
Publisher Copyright:
© Mary Ann Liebert, Inc.
PY - 2024/8
Y1 - 2024/8
N2 - Choroideremia (CHM) is a rare X-linked chorioretinal dystrophy causing progressive vision loss due to mutations in the CHM gene, leading to Rab escort protein 1 loss of function. CHM disease is characterized by a progressive degeneration of the choroid, the retinal pigment epithelium (RPE), and the retina. The RPE is a monolayer of polarized cells that supports photoreceptors, providing nutrients, growth factors, and ions, and removes retinal metabolism waste products, having a central role in CHM pathogenesis. Commonly used models such as ARPE-19 cells do not reproduce accurately the nature of RPE cells. Human induced pluripotent stem cells (hiPSCs) can be differentiated into RPE cells (hiPSC-RPE), which mimic key features of native RPE, being more suited to study retinal diseases. Therefore, we took advantage of hiPSCs to generate new human-based CHM models. Two isogenic hiPSC lines were generated through CRISPR/Cas9: a CHM knock-out line from a healthy donor and a corrected CHM patient line using a knock-in approach. The differentiated hiPSC-RPE lines exhibited critical morphological and physiological characteristics of native RPE, including the presence of the tight junction markers Claudin-19 and Zonula Occludens-1, phagocytosis of photoreceptor outer segments, pigmentation, a postmitotic state, and the characteristic polygonal shape. In addition, all the studied cells were able to form retinal organoids. This work resulted in the establishment of isogenic hiPSC lines, representing a new and important CHM cellular model. To our knowledge, this is the first time that isogenic cell lines have been developed to model CHM disease, providing a valuable tool for studying the mechanisms at the onset of RPE degeneration.
AB - Choroideremia (CHM) is a rare X-linked chorioretinal dystrophy causing progressive vision loss due to mutations in the CHM gene, leading to Rab escort protein 1 loss of function. CHM disease is characterized by a progressive degeneration of the choroid, the retinal pigment epithelium (RPE), and the retina. The RPE is a monolayer of polarized cells that supports photoreceptors, providing nutrients, growth factors, and ions, and removes retinal metabolism waste products, having a central role in CHM pathogenesis. Commonly used models such as ARPE-19 cells do not reproduce accurately the nature of RPE cells. Human induced pluripotent stem cells (hiPSCs) can be differentiated into RPE cells (hiPSC-RPE), which mimic key features of native RPE, being more suited to study retinal diseases. Therefore, we took advantage of hiPSCs to generate new human-based CHM models. Two isogenic hiPSC lines were generated through CRISPR/Cas9: a CHM knock-out line from a healthy donor and a corrected CHM patient line using a knock-in approach. The differentiated hiPSC-RPE lines exhibited critical morphological and physiological characteristics of native RPE, including the presence of the tight junction markers Claudin-19 and Zonula Occludens-1, phagocytosis of photoreceptor outer segments, pigmentation, a postmitotic state, and the characteristic polygonal shape. In addition, all the studied cells were able to form retinal organoids. This work resulted in the establishment of isogenic hiPSC lines, representing a new and important CHM cellular model. To our knowledge, this is the first time that isogenic cell lines have been developed to model CHM disease, providing a valuable tool for studying the mechanisms at the onset of RPE degeneration.
KW - choroideremia
KW - CRISPR/Cas9
KW - human induced pluripotent stem cells
KW - retinal pigment epithelium
UR - http://www.scopus.com/inward/record.url?scp=85201771966&partnerID=8YFLogxK
U2 - 10.1089/scd.2024.0105
DO - 10.1089/scd.2024.0105
M3 - Article
C2 - 39078329
AN - SCOPUS:85201771966
SN - 1547-3287
JO - Stem Cells and Development
JF - Stem Cells and Development
ER -