Nutraceutical‐mediated restoration of wild‐type levels of <i><scp>IKBKAP</scp></i>‐encoded <scp>IKAP</scp> protein in familial dysautonomia‐derived cells

Anderson, Sylvia L.; Liu, Bo; Qiu, Jinsong; Sturm, Andrea J.; Schwartz, Jamie A.; Peters, Austin; Sullivan, Kerry A.; Rubin, Berish Y.

doi:10.1002/mnfr.201100670

Cited by 11 publications

(5 citation statements)

References 56 publications

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“…C). In the light of our recent report demonstrating the ability of genistein to promote the inclusion of exon 4 in the CLK1 transcript , we examined the impact of genistein on the transcripts generated from these mutated constructs. A clear genistein‐mediated enhancement of exon 4 inclusion was observed, demonstrating that, while the base changes spanning nucleotides 829–838 disrupt the sequence required for digoxin responsiveness, these changes have no effect on the genistein‐mediated response (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, we recently reported that the isoflavones genistein and daidzein, which are found in soy, also modulate the splicing of the IKBKAP ‐derived transcript in FD‐derived cells, resulting in the enhanced production of the wild‐type, exon‐20‐containing, transcript from the IKBKAP gene bearing the FD‐causing mutation . We further showed that treatment of FD‐derived cells with a combination of genistein and EGCG induces cellular levels of IKAP protein that are indistinguishable from those produced in normal cells.…”

Section: Introductionmentioning

confidence: 99%

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Cardiac glycosides correct aberrant splicing of IKBKAP‐encoded mRNA in familial dysautonomia derived cells by suppressing expression of SRSF3

et al. 2013

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The ability to modulate the production of the wild-type transcript in cells bearing the splice-altering familial dysautonomia (FD) causing mutation in the IKBKAP gene prompted a study of the impact of a panel of pharmaceuticals on the splicing of this transcript, which revealed the ability of the cardiac glycoside digoxin to increase the production of the wild-type, exon-20-containing, IKBKAP-encoded transcript and the full-length IjB-kinasecomplex-associated protein in FD-derived cells. Characterization of the cis elements and trans factors involved in the digoxin-mediated effect on splicing reveals that this response is dependent on an SRSF3 binding site(s) located in the intron 5′ of the alternatively spliced exon and that digoxin mediates its effect by suppressing the level of the SRSF3 protein. Characterization of the digoxin-mediated effect on the RNA splicing process was facilitated by the identification of several RNA splicing events in which digoxin treatment mediates the enhanced inclusion of exonic sequence. Moreover, we demonstrate the ability of digoxin to impact the splicing process in neuronal cells, a cell type profoundly impacted by FD. This study represents the first demonstration that digoxin possesses splice-altering capabilities that are capable of reversing the impact of the FD-causing mutation. These findings support the clinical evaluation of the impact of digoxin on the FD patient population.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cardiac glycosides correct aberrant splicing of IKBKAP‐encoded mRNA in familial dysautonomia derived cells by suppressing expression of SRSF3

et al. 2013

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“…Most attempts to develop FD therapeutics have focused on correcting the splicing defect by increasing the levels of exon 20 inclusion through treatment with various small molecules, with varying degrees of success. Notable among these small molecules are rectifier of aberrant mRNA splicing (RECTAS) ( 13 ), epigallocatechin gallate ( 17 ), protease inhibitors ( 18 ), phosphatidylserine ( 5 , 19 , 20 ), tocotrienols ( 21 ), and kinetin ( 22 , 23 ). In clinical trials, tocotrienols did not demonstrate significant clinical efficacy ( 21 ), whereas kinetin administered orally moderately improved IKBKAP splicing in the white blood cells of the treated patients ( 22 ).…”

Section: Introductionmentioning

confidence: 99%

Antisense oligonucleotides correct the familial dysautonomia splicing defect in IKBKAP transgenic mice

Sinha

Kim

Nomakuchi

et al. 2018

Nucleic Acids Research

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Familial dysautonomia (FD) is a rare inherited neurodegenerative disorder caused by a point mutation in the IKBKAP gene that results in defective splicing of its pre-mRNA. The mutation weakens the 5′ splice site of exon 20, causing this exon to be skipped, thereby introducing a premature termination codon. Though detailed FD pathogenesis mechanisms are not yet clear, correcting the splicing defect in the relevant tissue(s), thus restoring normal expression levels of the full-length IKAP protein, could be therapeutic. Splice-switching antisense oligonucleotides (ASOs) can be effective targeted therapeutics for neurodegenerative diseases, such as nusinersen (Spinraza), an approved drug for spinal muscular atrophy. Using a two-step screen with ASOs targeting IKBKAP exon 20 or the adjoining intronic regions, we identified a lead ASO that fully restored exon 20 splicing in FD patient fibroblasts. We also characterized the corresponding cis-acting regulatory sequences that control exon 20 splicing. When administered into a transgenic FD mouse model, the lead ASO promoted expression of full-length human IKBKAP mRNA and IKAP protein levels in several tissues tested, including the central nervous system. These findings provide insights into the mechanisms of IKBKAP exon 20 recognition, and pre-clinical proof of concept for an ASO-based targeted therapy for FD.

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“…Remarkably, an antioxidant flavonoid named epigallocatechin gallate (EGCG), which is the most abundant and bioactive polyphenol in green tea, has been identified by the Rubin group as capable of correcting IKBKAP mRNA aberrant splicing by decreasing hnRNP A2/B1 expression level (Anderson et al, 2012;Anderson et al, 2003a). Although its efficacy in FD remains controversial (Lee et al, 2009), its anticancer properties have been in part related to proteasome inhibition (Chen et al, 2011;Modernelli et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Proteasome inhibitors to alleviate aberrant IKBKAP mRNA splicing and low IKAP/hELP1 synthesis in familial dysautonomia

Hervé

Ibrahim

2017

Neurobiology of Disease

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FD is a rare neurodegenerative disorder caused by a mutation of the IKBKAP gene, which induces low expression levels of the Elongator subunit IKAP/hELP1 protein. A rational strategy for FD treatment could be to identify drugs increasing IKAP/hELP1 expression levels by blocking protein degradation pathways such as the 26S proteasome. Proteasome inhibitors are promising molecules emerging in cancer treatment and could thus constitute an enticing pharmaceutical strategy for FD treatment. Therefore, we tested three proteasome inhibitors on FD human olfactory ecto-mesenchymal stem cells (hOE-MSCs): two approved by the Food and Drug Administration (FDA) and European Medicines Agency (EMA), bortezomib and carfilzomib, as well as epoxomicin. Although all 3 inhibitors demonstrated activity in correcting IKBKAP mRNA aberrant splicing, carfilzomib was superior in enhancing IKAP/hELP1 quantity. Moreover, we observed a synergistic effect of suboptimal doses of carfilzomib on kinetin in improving IKBKAP isoforms ratio and IKAP/hELP1 expression levels allowing to counterbalance carfilzomib toxicity. Finally, we identified several dysregulated miRNAs after carfilzomib treatment that target proteasome-associated mRNAs and determined that IKAP/hELP1 deficiency in FD pathology is correlated to an overactivity of the 26S proteasome. Altogether, these results reinforce the rationale for using chemical compounds inhibiting the 26S proteasome as an innovative option for FD and a promising therapeutic pathway for many other neurodegenerative diseases.

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Nutraceutical‐mediated restoration of wild‐type levels of IKBKAP‐encoded IKAP protein in familial dysautonomia‐derived cells

Cited by 11 publications

References 56 publications

Cardiac glycosides correct aberrant splicing of IKBKAP‐encoded mRNA in familial dysautonomia derived cells by suppressing expression of SRSF3

Cardiac glycosides correct aberrant splicing of IKBKAP‐encoded mRNA in familial dysautonomia derived cells by suppressing expression of SRSF3

Antisense oligonucleotides correct the familial dysautonomia splicing defect in IKBKAP transgenic mice

Proteasome inhibitors to alleviate aberrant IKBKAP mRNA splicing and low IKAP/hELP1 synthesis in familial dysautonomia

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