Patient-specific induced pluripotent stem cell-derived RPE cells

Understanding the pathogenesis of retinopathy in long-chain 3-hydroxyacyl-CoA dehydrogenase deiciency

Padmini P. Polinati, Tanja Ilmarinen, Ras Trokovic, Tuulia Hyotylainen, Timo Otonkoski, Anu Suomalainen, Heli Skottman, Tiina Tyni

Research output: Contribution to journalArticleScientificpeer-review

12 Citations (Scopus)

Abstract

Purpose. Retinopathy is an important manifestation of trifunctional protein (TFP) deficiencies but not of other defects of fatty acid oxidation. The common homozygous mutation in the TFP a-subunit gene HADHA (hydroxyacyl-CoA dehydrogenase), c.1528G>C, affects the long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) activity of TFP and blindness in infancy. The pathogenesis of the retinopathy is unknown. This study aimed to utilize human induced pluripotent stem cell (hiPSC) technology to create a disease model for the disorder, and to derive clues for retinopathy pathogenesis. Methods. We implemented hiPSC technology to generate LCHAD deficiency (LCHADD) patient-specific retinal pigment epithelial (RPE) monolayers. These patient and control RPEs were extensively characterized for function and structure, as well as for lipid composition by mass spectrometry. Results. The hiPSC-derived RPE monolayers of patients and controls were functional, as they both were able to phagocytose the photoreceptor outer segments in vitro. Interestingly, the patient RPEs had intense cytoplasmic neutral lipid accumulation, and lipidomic analysis revealed an increased triglyceride accumulation. Further, patient RPEs were small and irregular in shape, and their tight junctions were disorganized. Their ultratructure showed decreased pigmentation, few melanosomes, and more melanolysosomes. Conclusions. We demonstrate that the RPE cell model reveals novel early pathogenic changes in LCHADD retinopathy, with robust lipid accumulation, inefficient pigmentation that is evident soon after differentiation, and a defect in forming tight junctions inducing apoptosis. We propose that LCHADD-RPEs are an important model for mitochondrial TFP retinopathy, and that their early pathogenic changes contribute to infantile blindness of LCHADD.
Original languageEnglish
Pages (from-to)3371-3382
JournalInvestigative Ophthalmology and Visual Science
Volume56
Issue number5
DOIs
Publication statusPublished - 2015
MoE publication typeA1 Journal article-refereed

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3-Hydroxyacyl-CoA Dehydrogenase
Induced Pluripotent Stem Cells
Retinal Pigments
Epithelial Cells
Tight Junctions
Pigmentation
Blindness
Lipids
Mitochondrial Trifunctional Protein
Technology
Melanosomes
Protein Subunits
Coenzyme A
Phagocytosis
Mass Spectrometry
Oxidoreductases
Triglycerides
Fatty Acids
Apoptosis
Mutation

Keywords

  • beta oxidation
  • mitochondria
  • retinal pigment epithelium
  • retinopathy

Cite this

Polinati, Padmini P. ; Ilmarinen, Tanja ; Trokovic, Ras ; Hyotylainen, Tuulia ; Otonkoski, Timo ; Suomalainen, Anu ; Skottman, Heli ; Tyni, Tiina. / Patient-specific induced pluripotent stem cell-derived RPE cells : Understanding the pathogenesis of retinopathy in long-chain 3-hydroxyacyl-CoA dehydrogenase deiciency. In: Investigative Ophthalmology and Visual Science. 2015 ; Vol. 56, No. 5. pp. 3371-3382.
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title = "Patient-specific induced pluripotent stem cell-derived RPE cells: Understanding the pathogenesis of retinopathy in long-chain 3-hydroxyacyl-CoA dehydrogenase deiciency",
abstract = "Purpose. Retinopathy is an important manifestation of trifunctional protein (TFP) deficiencies but not of other defects of fatty acid oxidation. The common homozygous mutation in the TFP a-subunit gene HADHA (hydroxyacyl-CoA dehydrogenase), c.1528G>C, affects the long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) activity of TFP and blindness in infancy. The pathogenesis of the retinopathy is unknown. This study aimed to utilize human induced pluripotent stem cell (hiPSC) technology to create a disease model for the disorder, and to derive clues for retinopathy pathogenesis. Methods. We implemented hiPSC technology to generate LCHAD deficiency (LCHADD) patient-specific retinal pigment epithelial (RPE) monolayers. These patient and control RPEs were extensively characterized for function and structure, as well as for lipid composition by mass spectrometry. Results. The hiPSC-derived RPE monolayers of patients and controls were functional, as they both were able to phagocytose the photoreceptor outer segments in vitro. Interestingly, the patient RPEs had intense cytoplasmic neutral lipid accumulation, and lipidomic analysis revealed an increased triglyceride accumulation. Further, patient RPEs were small and irregular in shape, and their tight junctions were disorganized. Their ultratructure showed decreased pigmentation, few melanosomes, and more melanolysosomes. Conclusions. We demonstrate that the RPE cell model reveals novel early pathogenic changes in LCHADD retinopathy, with robust lipid accumulation, inefficient pigmentation that is evident soon after differentiation, and a defect in forming tight junctions inducing apoptosis. We propose that LCHADD-RPEs are an important model for mitochondrial TFP retinopathy, and that their early pathogenic changes contribute to infantile blindness of LCHADD.",
keywords = "beta oxidation, mitochondria, retinal pigment epithelium, retinopathy",
author = "Polinati, {Padmini P.} and Tanja Ilmarinen and Ras Trokovic and Tuulia Hyotylainen and Timo Otonkoski and Anu Suomalainen and Heli Skottman and Tiina Tyni",
year = "2015",
doi = "10.1167/iovs.14-14007",
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Patient-specific induced pluripotent stem cell-derived RPE cells : Understanding the pathogenesis of retinopathy in long-chain 3-hydroxyacyl-CoA dehydrogenase deiciency. / Polinati, Padmini P.; Ilmarinen, Tanja; Trokovic, Ras; Hyotylainen, Tuulia; Otonkoski, Timo; Suomalainen, Anu; Skottman, Heli; Tyni, Tiina.

In: Investigative Ophthalmology and Visual Science, Vol. 56, No. 5, 2015, p. 3371-3382.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Patient-specific induced pluripotent stem cell-derived RPE cells

T2 - Understanding the pathogenesis of retinopathy in long-chain 3-hydroxyacyl-CoA dehydrogenase deiciency

AU - Polinati, Padmini P.

AU - Ilmarinen, Tanja

AU - Trokovic, Ras

AU - Hyotylainen, Tuulia

AU - Otonkoski, Timo

AU - Suomalainen, Anu

AU - Skottman, Heli

AU - Tyni, Tiina

PY - 2015

Y1 - 2015

N2 - Purpose. Retinopathy is an important manifestation of trifunctional protein (TFP) deficiencies but not of other defects of fatty acid oxidation. The common homozygous mutation in the TFP a-subunit gene HADHA (hydroxyacyl-CoA dehydrogenase), c.1528G>C, affects the long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) activity of TFP and blindness in infancy. The pathogenesis of the retinopathy is unknown. This study aimed to utilize human induced pluripotent stem cell (hiPSC) technology to create a disease model for the disorder, and to derive clues for retinopathy pathogenesis. Methods. We implemented hiPSC technology to generate LCHAD deficiency (LCHADD) patient-specific retinal pigment epithelial (RPE) monolayers. These patient and control RPEs were extensively characterized for function and structure, as well as for lipid composition by mass spectrometry. Results. The hiPSC-derived RPE monolayers of patients and controls were functional, as they both were able to phagocytose the photoreceptor outer segments in vitro. Interestingly, the patient RPEs had intense cytoplasmic neutral lipid accumulation, and lipidomic analysis revealed an increased triglyceride accumulation. Further, patient RPEs were small and irregular in shape, and their tight junctions were disorganized. Their ultratructure showed decreased pigmentation, few melanosomes, and more melanolysosomes. Conclusions. We demonstrate that the RPE cell model reveals novel early pathogenic changes in LCHADD retinopathy, with robust lipid accumulation, inefficient pigmentation that is evident soon after differentiation, and a defect in forming tight junctions inducing apoptosis. We propose that LCHADD-RPEs are an important model for mitochondrial TFP retinopathy, and that their early pathogenic changes contribute to infantile blindness of LCHADD.

AB - Purpose. Retinopathy is an important manifestation of trifunctional protein (TFP) deficiencies but not of other defects of fatty acid oxidation. The common homozygous mutation in the TFP a-subunit gene HADHA (hydroxyacyl-CoA dehydrogenase), c.1528G>C, affects the long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) activity of TFP and blindness in infancy. The pathogenesis of the retinopathy is unknown. This study aimed to utilize human induced pluripotent stem cell (hiPSC) technology to create a disease model for the disorder, and to derive clues for retinopathy pathogenesis. Methods. We implemented hiPSC technology to generate LCHAD deficiency (LCHADD) patient-specific retinal pigment epithelial (RPE) monolayers. These patient and control RPEs were extensively characterized for function and structure, as well as for lipid composition by mass spectrometry. Results. The hiPSC-derived RPE monolayers of patients and controls were functional, as they both were able to phagocytose the photoreceptor outer segments in vitro. Interestingly, the patient RPEs had intense cytoplasmic neutral lipid accumulation, and lipidomic analysis revealed an increased triglyceride accumulation. Further, patient RPEs were small and irregular in shape, and their tight junctions were disorganized. Their ultratructure showed decreased pigmentation, few melanosomes, and more melanolysosomes. Conclusions. We demonstrate that the RPE cell model reveals novel early pathogenic changes in LCHADD retinopathy, with robust lipid accumulation, inefficient pigmentation that is evident soon after differentiation, and a defect in forming tight junctions inducing apoptosis. We propose that LCHADD-RPEs are an important model for mitochondrial TFP retinopathy, and that their early pathogenic changes contribute to infantile blindness of LCHADD.

KW - beta oxidation

KW - mitochondria

KW - retinal pigment epithelium

KW - retinopathy

U2 - 10.1167/iovs.14-14007

DO - 10.1167/iovs.14-14007

M3 - Article

VL - 56

SP - 3371

EP - 3382

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 5

ER -