Restoring Ciliary Function: Gene Therapeutics for Primary Ciliary Dyskinesia

Keiser, Nicholas W.; Cant, Erin; Sitaraman, Sneha; Shoemark, Amelia; Limberis, Maria P.

doi:10.1089/hum.2023.102

Cited by 6 publications

(1 citation statement)

References 163 publications

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“…Additionally, our study focused on nonmotile cilia and it would be of interest to see if MI-181 also translates to improving cilia length and/or function of motile cilia, which could address respiratory illnesses like mucociliary diseases that are characterized by defective motile cilia. 22 ■ METHODS Cell Culture. hTERT RPE-1 cells (ATCC, verified by shorttandem repeat profiling) were cultured in DMEM/F12 1:1 (Cytiva) with 10% FBS (Gibco) in 5% CO 2 at 37 °C.…”

Section: ■ Conclusionmentioning

confidence: 99%

MI-181 Modulates Cilia Length and Restores Cilia Length in Cells with Defective Shortened Cilia

Gholkar,

Gimeno,

Edgemon

et al. 2024

ACS Chem. Biol.

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Primary cilia are membrane-covered microtubule-based structures that protrude from the cell surface and are critical for cell signaling and homeostasis during human development and adulthood. Dysregulation of cilia formation, length, and function can lead to a spectrum of human diseases and syndromes known as ciliopathies. Although some genetic and chemical screens have been performed to define important factors that modulate cilia biogenesis and length control, there are currently no clinical treatments that restore cilia length in patients. We report that the microtubule-targeting agent MI-181(mitotic inhibitor-181) is a potent modulator of cilia length and biogenesis. Treatment of retinal pigment epithelial-1 cells with MI-181 induced an increase in the average size of cilia and in the percent ciliated cells under nonstarved conditions. Importantly, MI-181 was effective at rescuing cilia length and ciliation defects in cells that had been treated with the intraflagellar transport inhibitor Ciliobrevin D or the O-GlcNAc transferase inhibitor OSMI-1. Most importantly, MI-181 induced an increase in cilia length and restored ciliation in cells with compromised shortened cilia at low nanomolar concentrations and did not show an inhibitory response at high concentrations. Therefore, MI-181 represents a lead molecule for developing drugs targeting ciliopathies characterized by shortened cilia.

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Section: ■ Conclusionmentioning

confidence: 99%

MI-181 Modulates Cilia Length and Restores Cilia Length in Cells with Defective Shortened Cilia

Gholkar,

Gimeno,

Edgemon

et al. 2024

ACS Chem. Biol.

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Promoter Deletion Leading to Allele Specific Expression in a Genetically Unsolved Case of Primary Ciliary Dyskinesia

Beaman,

Yin,

Smith

et al. 2024

American J of Med Genetics Pt A

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Variation in the non‐coding genome represents an understudied mechanism of disease and it remains challenging to predict if single nucleotide variants, small insertions and deletions, or structural variants in non‐coding genomic regions will be detrimental. Our approach using complementary RNA‐seq and targeted long‐read DNA sequencing can prioritize identification of non‐coding variants that lead to disease via alteration of gene splicing or expression. We have identified a patient with primary ciliary dyskinesia with a pathogenic coding variant on one allele of the SPAG1 gene, while the second allele appears normal by whole exome sequencing despite an autosomal recessive inheritance pattern. RNA sequencing revealed reduced SPAG1 transcript levels and exclusive allele specific expression of the known pathogenic allele, suggesting the presence of a non‐coding variant on the second allele that impacts transcription. Targeted long‐read DNA sequencing identified a heterozygous 3 kilobase deletion of the 5′ untranslated region of SPAG1, overlapping the promoter and first non‐coding exon. This non‐coding deletion was missed by whole exome sequencing and gene‐specific deletion/duplication analysis, highlighting the importance of investigating the non‐coding genome in patients with “missing” disease‐causing variation. This paradigm demonstrates the utility of both RNA and long‐read DNA sequencing in identifying pathogenic non‐coding variants in patients with unexplained genetic disease.

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