Background Leber’s inherited optic neuropathy (LHON) is well known for incomplete penetrance. A pair of monozygotic twins carrying 14484T > C LHON mutation: one displayed LHON characteristics (affected LHON) and the other twin was an unaffected LHON carrier, were studied to identify possible modifier(s) for LHON manifestation. Methods Primary fibroblasts from affected and unaffected monozygotic twins with 14484T > C LHON mutation were treated with different insults to differentiate cellular phenotype between the two fibroblasts. RNA sequencing of the fibroblasts indicated differentially expressed genes and whole genome sequencing was used to identify candidates for disease modifier. Results Our results suggested that fibroblast from unaffected carrier was able to adapt to galactose and hydrogen peroxide insult, while affected fibroblasts were not. We found reduced expression of total SOD2 with high proportion of inactive SOD2 (acetylated SOD2) in affected LHON fibroblast, while decreased expression of SIRT3 was detected in affected LHON fibroblasts treated with combination of insults. Differential expression indicated enrichment of a pathway relating to negative regulator of cell death pathway in unaffected carrier fibroblast. Expression of receptor for prostaglandin E receptor (PTGER3) was found to be affected by two SNPs. Unaffected LHON fibroblast possessed rs75523942 that indicates a positive effect on PTGER3 expression, while affected LHON fibroblast possessed rs496483 that indicates a negative effect on PTGER3 expression. Discordant SNPs on prostaglandin E receptor 3 (PTGER3) were identified as eQTL. Conclusions This study indicates that prostanoid receptor could be a possible modifier for LHON manifestation of these twins.

Spinal muscular atrophy (SMA) is a severe, debilitating neuromuscular condition characterised by loss of motor neurons and progressive muscle wasting. SMA is caused by a loss of expression of SMN1 that encodes the survival motor neuron (SMN) protein necessary for the survival of motor neurons. Restoration of SMN expression through increased inclusion of SMN2 exon 7 is known to ameliorate symptoms in SMA patients. As a consequence, regulation of pre-mRNA splicing of SMN2 could provide a potential molecular therapy for SMA. In this study, we explored if splice switching antisense oligonucleotides could redirect the splicing repressor hnRNPA1 to the hnRNPA1b isoform and restore SMN expression in fibroblasts from a type I SMA patient. Antisense oligonucleotides (AOs) were designed to promote exon 7b retention in the mature mRNA and induce the hnRNPA1b isoform. RT-PCR and western blot analysis were used to assess and monitor the efficiency of different AO combinations. A combination of AOs targeting multiple silencing motifs in hnRNPA1 pre-mRNA led to robust hnRNPA1b induction, which, in turn, significantly increased expression of full-length SMN (FL-SMN) protein. A combination of PMOs targeting the same motifs also strongly induced hnRNPA1b isoform, but surprisingly SMN2 exon 5 skipping was detected, and the PMO cocktail did not lead to a significant increase in expression of FL-SMN protein. We further performed RNA sequencing to assess the genome-wide effects of hnRNPA1b induction. Some 3244 genes were differentially expressed between the hnRNPA1b-induced and untreated SMA fibroblasts, which are functionally enriched in cell cycle and chromosome segregation processes. RT-PCR analysis demonstrated that expression of the master regulator of these enrichment pathways, MYBL2 and FOXM1B, were reduced in response to PMO treatment. These findings suggested that induction of hnRNPA1b can promote SMN protein expression, but not at sufficient levels to be clinically relevant.