Exploiting hiPSCs in Leber's Hereditary Optic Neuropathy (LHON): Present Achievements and Future Perspectives

Peron, Camille; Maresca, Alessandra; Cavaliere, Andrea; Iannielli, Angelo; Broccoli, Vania; Carelli, Valerio; Meo, Ivano Di; Tiranti, Valeria

doi:10.3389/fneur.2021.648916

Cited by 8 publications

(3 citation statements)

References 88 publications

(102 reference statements)

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“…However, some limitations still exist. For example, the clone number was insufficient after differentiation due to immature technology ( Peron et al, 2021 ), and mutated mtDNA might be missed in partial RGCs differentiated from iPSC{Peron, 2021 #79}. Skin-derived fibroblasts are a proven alternative cell model ( Jankauskaite et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

Superoxide dismutase 2 ameliorates mitochondrial dysfunction in skin fibroblasts of Leber’s hereditary optic neuropathy patients

et al. 2022

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BackgroundIn Leber’s hereditary optic neuropathy (LHON), mtDNA mutations mediate mitochondrial dysfunction and apoptosis of retinal ganglion cells. Mitochondrial superoxide dismutase 2 (SOD2) is a crucial antioxidase against reactive oxygen species (ROS). This study aims to investigate whether SOD2 could ameliorate mtDNA mutation mediated mitochondrial dysfunction in skin fibroblasts of LHON patients and explore the underlying mechanisms.MethodsThe skin of normal healthy subjects and severe LHON patients harboring m.11778G > A mutation was taken to prepare immortalized skin fibroblast cell lines (control-iFB and LHON-iFB). LHON-iFB cells were transfected with SOD2 plasmid or negative control plasmid, respectively. In addition, human neuroblastoma SH-SY5Y cells and human primary retinal pigmental epithelium (hRPE) cells were stimulated by H2O2 after gene transfection. The oxygen consumption rate (OCR) was measured with a Seahorse extracellular flux analyzer. The level of ATP production, mitochondrial membrane potential, ROS and malondialdehyde (MDA) were measured separately with the corresponding assay kits. The expression level of SOD2, inflammatory cytokines and p-IκBα/IκBα was evaluated by western-blot. Assessment of apoptosis was performed by TUNEL assay.ResultsLHON-iFB exhibited lower OCR, ATP production, mitochondrial membrane potential but higher level of ROS and MDA than control-iFB. Western-blot revealed a significantly increased expression of IL-6 and p-IκBα/IκBα in LHON-iFB. Compared with the negative control, SOD2 overexpression increased OCR, ATP production and elevated mitochondrial membrane potential, but impaired ROS and MDA production. Besides, western-blot demonstrated exogenous SOD2 reduced the protein level of IL-6 and p-IκBα/IκBα. TUNEL assays suggested SOD2 inhibited cells apoptosis. Analogously, in SH-SY5Y and hRPE cells, SOD2 overexpression increased ATP production and mitochondrial membrane potential, but decreased ROS, MDA levels and suppressed apoptosis.ConclusionSOD2 upregulation inhibited cells apoptosis through ameliorating mitochondrial dysfunction and reducing NF-κB associated inflammatory response. This study further support exogenous SOD2 may be a promising therapy for the treatment of LHON.

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Section: Discussionmentioning

confidence: 99%

Superoxide dismutase 2 ameliorates mitochondrial dysfunction in skin fibroblasts of Leber’s hereditary optic neuropathy patients

et al. 2022

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“…The iPSCs-derived RGCs from patients carrying m.11778G > A mutation displayed defective mitochondrial biogenesis, decreased basal respiration, increased oxidative stress and increased apoptosis ( 28 , 29 , 33 ). Here, the RGC-like cells bearing only m.11778G > A or p.Arg53Trp mutation exhibited mild decreases in the mitochondrial ATP contents, comparable with those in fibroblast, lymphoblastoid or cybrid cell lines ( 17 , 20 , 25 ).…”

Section: Discussionmentioning

confidence: 99%

“…The use of human-induced pluripotent stem cells (hiPSCs) derived from patients to obtain terminally differentiated RGCs and neurons is a revolutionary approach to understanding pathogenic mechanisms of retinal disorders ( 28–34 ). To further elucidate the pathophysiology of LHON-linked m.11778G > A and PRICKLE3 p.Arg53Trp mutations, we generated the induced pluripotent stem cells (iPSCs) derived from matrilineal relatives of one Chinese family (asymptomatic carriers carrying only the m.11778G > A mutation or PRICKLE3 p.Arg53Trp mutation, symptomatic individuals bearing both m.11778G > A and heterozygous or hemizygous PRICKLE3 p.Arg53Trp mutations), and married-in control lacking these mutations ( 25 , 30 , 31 ).…”

Section: Introductionmentioning

confidence: 99%

Abnormal morphology and function in retinal ganglion cells derived from patients-specific iPSCs generated from individuals with Leber’s hereditary optic neuropathy

Nie

Wang

Chen

et al. 2022

Human Molecular Genetics

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Leber’s hereditary optic neuropathy (LHON) is a maternally inherited eye disease that results from degeneration of retinal ganglion cells (RGC). Mitochondrial ND4 11778G > A mutation, which affects structural components of complex I, is the most prevalent LHON-associated mitochondrial DNA (mtDNA) mutation worldwide. The m.11778G > A mutation is the primary contributor underlying the development of LHON and X-linked PRICKLE3 allele (c.157C > T, p.Arg53Trp) linked to biogenesis of ATPase interacts with m.11778G > A mutation to cause LHON. However, the lack of appropriate cell and animal models of LHON has been significant obstacles for deep elucidation of disease pathophysiology, specifically the tissue specific effects. Using RGC-like cells differentiated from induced pluripotent stem cells (iPSCs) from members of one Chinese family (asymptomatic subjects carrying only m.11778G > A mutation or PRICKLE3 p.Arg53Trp mutation, symptomatic individuals bearing both m.11778G > A and PRICKLE3 p.Arg53Trp mutations and control lacking these mutations), we demonstrated the deleterious effects of mitochondrial dysfunctions on the morphology and functions of RGCs. Notably, iPSCs bearing only m.11778G > A or p.Arg53Trp mutation exhibited mild defects in differentiation to RGC-like cells. The RGC-like cells carrying only m.11778G > A or p.Arg53Trp mutation displayed mild defects in RGC morphology, including the area of soma and numbers of neurites, electrophysiological properties, ATP production and apoptosis. Strikingly, those RGC-like cells derived from symptomatic individuals harboring both m.11778G > A and p.Arg53Trp mutations displayed greater defects in the development, morphology and functions than those in cells bearing single mutation. These findings provide new insights into pathophysiology of LHON arising from RGC deficiencies caused by synergy between m.11778G > A and PRICKLE3 p.Arg53Trp mutation.

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Leveraging neural crest pluripotency to extend retinal and craniofacial niches for building neurovascular organoids—a theranostic and drug development perspective

Sharma¹,

Jangra²,

Dhiman³

et al. 2023

The Eye, Volume 4

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Exploiting hiPSCs in Leber's Hereditary Optic Neuropathy (LHON): Present Achievements and Future Perspectives

Cited by 8 publications

References 88 publications

Superoxide dismutase 2 ameliorates mitochondrial dysfunction in skin fibroblasts of Leber’s hereditary optic neuropathy patients

Superoxide dismutase 2 ameliorates mitochondrial dysfunction in skin fibroblasts of Leber’s hereditary optic neuropathy patients

Abnormal morphology and function in retinal ganglion cells derived from patients-specific iPSCs generated from individuals with Leber’s hereditary optic neuropathy

Leveraging neural crest pluripotency to extend retinal and craniofacial niches for building neurovascular organoids—a theranostic and drug development perspective

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