Gene editing of<i>DNAH11</i>restores normal cilia motility in primary ciliary dyskinesia

Lai, Michele; Pifferi, Massimo; Bush, Andrew; Piras, Martina; Michelucci, Angela; Cicco, Maria Di; Grosso, Ambra Del; Quaranta, Paola; Cursi, Chiara; Tantillo, Elena; Franceschi, Sara; Mazzanti, M. C.; Simi, Paolo; Saggese, Giuseppe; Boner, Attilio; Pistello, Mauro

doi:10.1136/jmedgenet-2015-103539

Cited by 53 publications

(37 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This vector contains Blue Fluorescent Protein (BFP) and, as outlined above, gRNA targeting exon 6. We have used a multiplicity of infection (MOI) of 1, as described 28 . The A375 cell line was first transduced with the lentiviral vector.…”

Section: Methodsmentioning

confidence: 99%

Complete Acid Ceramidase ablation prevents cancer-initiating cell formation in melanoma cells

Lai

Realini

Ferla

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

Acid ceramidase (AC) is a lysosomal cysteine hydrolase that catalyzes the conversion of ceramide into fatty acid and sphingosine. This reaction lowers intracellular ceramide levels and concomitantly generates sphingosine used for sphingosine-1-phosphate (S1P) production. Since increases in ceramide and consequent decreases of S1P reduce proliferation of various cancers, AC might offer a new target for anti-tumor therapy. Here we used CrispR-Cas9-mediated gene editing to delete the gene encoding for AC, ASAH1, in human A375 melanoma cells. ASAH1-null clones show significantly greater accumulation of long-chain saturated ceramides that are substrate for AC. As seen with administration of exogenous ceramide, AC ablation blocks cell cycle progression and accelerates senescence. Importantly, ASAH1-null cells also lose the ability to form cancer-initiating cells and to undergo self-renewal, which is suggestive of a key role for AC in maintaining malignancy and self-renewal of invasive melanoma cells. The results suggest that AC inhibitors might find therapeutic use as adjuvant therapy for advanced melanoma.

show abstract

Section: Methodsmentioning

confidence: 99%

Complete Acid Ceramidase ablation prevents cancer-initiating cell formation in melanoma cells

Lai

Realini

Ferla

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Targeted endonucleases create double-stranded breaks at specific points in the genome allowing for DNA repair, which can restore the wild-type genotype ( 43 ). Promising results have been achieved in cells from patients with the motile ciliopathy primary ciliary dyskinesia where ciliation was restored on replacement of the wild-type DNAH11 sequence ( 73 ). Preclinical work and clinical trials for other diseases using genome editing techniques are progressing with significant advances in animal models of epidermolysis bullosa ( 74 ).…”

Section: Future Therapies For Bbsmentioning

confidence: 99%

Managing Bardet–Biedl Syndrome—Now and in the Future

et al. 2018

View full text Add to dashboard Cite

Bardet–Biedl syndrome is a rare autosomal recessive multisystem disorder caused by defects in genes encoding for proteins that localize to the primary cilium/basal body complex. Twenty-one disease-causing genes have been identified to date. It is one of the most well-studied conditions in the family of diseases caused by defective cilia collectively known as ciliopathies. In this review, we provide an update on diagnostic developments, clinical features, and progress in the management of Bardet–Biedl syndrome. Advances in diagnostic technologies including exome and whole genome sequencing are expanding the spectrum of patients who are diagnosed with Bardet–Biedl syndrome and increasing the number of cases with diagnostic uncertainty. As a result of the diagnostic developments, a small number of patients with only one or two clinical features of Bardet–Biedl syndrome are being diagnosed. Our understanding of the syndrome-associated renal disease has evolved and is reviewed here. Novel interventions are developing at a rapid pace and are explored in this review including genetic therapeutics such as gene therapy, exon skipping therapy, nonsense suppression therapy, and gene editing. Other non-genetic therapies such as gene repurposing, targeted therapies, and non-pharmacological interventions are also discussed.

show abstract

“…A great expectation has originated from the recent study by Pifferi et al who first applied the “gene editing” to PCD; thus, they restored DNAH11 gene function ex vivo by replacing the inactivating mutation with wild-type sequence in the diseased cell (150). A new exciting era is cheerfully rising from genetic studies that will result in improving the outcome of affected patients.…”

Section: Current and Future Treatment Strategiesmentioning

confidence: 99%

Primary Ciliary Dyskinesia: An Update on Clinical Aspects, Genetics, Diagnosis, and Future Treatment Strategies

2017

View full text Add to dashboard Cite

Primary ciliary dyskinesia (PCD) is an orphan disease (MIM 244400), autosomal recessive inherited, characterized by motile ciliary dysfunction. The estimated prevalence of PCD is 1:10,000 to 1:20,000 live-born children, but true prevalence could be even higher. PCD is characterized by chronic upper and lower respiratory tract disease, infertility/ectopic pregnancy, and situs anomalies, that occur in ≈50% of PCD patients (Kartagener syndrome), and these may be associated with congenital heart abnormalities. Most patients report a daily year-round wet cough or nose congestion starting in the first year of life. Daily wet cough, associated with recurrent infections exacerbations, results in the development of chronic suppurative lung disease, with localized-to-diffuse bronchiectasis. No diagnostic test is perfect for confirming PCD. Diagnosis can be challenging and relies on a combination of clinical data, nasal nitric oxide levels plus cilia ultrastructure and function analysis. Adjunctive tests include genetic analysis and repeated tests in ciliary culture specimens. There are currently 33 known genes associated with PCD and correlations between genotype and ultrastructural defects have been increasingly demonstrated. Comprehensive genetic testing may hopefully screen young infants before symptoms occur, thus improving survival. Recent surprising advances in PCD genetic designed a novel approach called “gene editing” to restore gene function and normalize ciliary motility, opening up new avenues for treating PCD. Currently, there are no data from randomized clinical trials to support any specific treatment, thus, management strategies are usually extrapolated from cystic fibrosis. The goal of treatment is to prevent exacerbations, slowing the progression of lung disease. The therapeutic mainstay includes airway clearance maneuvers mainly with nebulized hypertonic saline and chest physiotherapy, and prompt and aggressive administration of antibiotics. Standardized care at specialized centers using a multidisciplinary approach that imposes surveillance of lung function and of airway biofilm composition likely improves patients’ outcome. Pediatricians, neonatologists, pulmonologists, and ENT surgeons should maintain high awareness of PCD and refer patients to the specialized center before sustained irreversible lung damage develops. The recent creation of a network of PCD clinical centers, focusing on improving diagnosis and treatment, will hopefully help to improve care and knowledge of PCD patients.

show abstract

Gene editing ofDNAH11restores normal cilia motility in primary ciliary dyskinesia

Abstract: This study demonstrates that gene editing can rescue ciliary beating ex vivo, opening up new avenues for treating PCD.

Cited by 53 publications

References 48 publications

Complete Acid Ceramidase ablation prevents cancer-initiating cell formation in melanoma cells

Complete Acid Ceramidase ablation prevents cancer-initiating cell formation in melanoma cells

Managing Bardet–Biedl Syndrome—Now and in the Future

Primary Ciliary Dyskinesia: An Update on Clinical Aspects, Genetics, Diagnosis, and Future Treatment Strategies

Contact Info

Product

Resources

About