The Scope for Thalassemia Gene Therapy by Disruption of Aberrant Regulatory Elements.
Petros PatsaliClaudio MussolinoPetros LadasArgyro FlogaAnnita KolnagouSoteroula ChristouMaria SitarouMichael N AntoniouToni CathomenCarsten Werner LedererMarina KleanthousPublished in: Journal of clinical medicine (2019)
The common IVSI-110 (G>A) β-thalassemia mutation is a paradigm for intronic disease-causing mutations and their functional repair by non-homologous end joining-mediated disruption. Such mutation-specific repair by disruption of aberrant regulatory elements (DARE) is highly efficient, but to date, no systematic analysis has been performed to evaluate disease-causing mutations as therapeutic targets. Here, DARE was performed in highly characterized erythroid IVSI-110(G>A) transgenic cells and the disruption events were compared with published observations in primary CD34+ cells. DARE achieved the functional correction of β-globin expression equally through the removal of causative mutations and through the removal of context sequences, with disruption events and the restriction of indel events close to the cut site closely resembling those seen in primary cells. Correlation of DNA-, RNA-, and protein-level findings then allowed the extrapolation of findings to other mutations by in silico analyses for potential repair based on the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9, Cas12a, and transcription activator-like effector nuclease (TALEN) platforms. The high efficiency of DARE and unexpected freedom of target design render the approach potentially suitable for 14 known thalassemia mutations besides IVSI-110(G>A) and put it forward for several prominent mutations causing other inherited diseases. The application of DARE, therefore, has a wide scope for sustainable personalized advanced therapy medicinal product development for thalassemia and beyond.
Keyphrases
- induced apoptosis
- crispr cas
- genome editing
- highly efficient
- cell cycle arrest
- gene therapy
- high efficiency
- sickle cell disease
- transcription factor
- endoplasmic reticulum stress
- genome wide
- systematic review
- poor prognosis
- randomized controlled trial
- inflammatory response
- binding protein
- immune response
- circulating tumor
- regulatory t cells
- cell proliferation
- toll like receptor
- cell therapy
- dendritic cells
- nuclear factor
- dna binding
- cell free
- replacement therapy
- nucleic acid