Effective viral-mediated lung gene therapy: is airway surface preparation necessary?

McCarron, Alexandra; Cmielewski, Patricia; Drysdale, Victoria; Parsons, David; Donnelley, Martin

doi:10.1038/s41434-022-00332-7

Cited by 18 publications

(7 citation statements)

References 108 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…cannula. To ensure that the tracheal airway tissue was receptive to LV transduction 27 , it was conditioned using our previously-described mechanical perturbation protocol, in which the tracheal airway surface was rubbed axially with a wire basket (N-Circle, Nitinol Tipless Stone Extractor NTSE-022115-UDH, Cook Medical, USA) for 30 s 28 . LV-MP tracheal dosing was then performed in a biosafety cabinet ~ 10 min after perturbation.…”

Section: Methodsmentioning

confidence: 99%

Improved in-vivo airway gene transfer via magnetic-guidance, with protocol development informed by synchrotron imaging

Donnelley

Cmielewski

Morgan

et al. 2022

Sci Rep

Self Cite

View full text Add to dashboard Cite

Gene vectors to treat cystic fibrosis lung disease should be targeted to the conducting airways, as peripheral lung transduction does not offer therapeutic benefit. Viral transduction efficiency is directly related to the vector residence time. However, delivered fluids such as gene vectors naturally spread to the alveoli during inspiration, and therapeutic particles of any form are rapidly cleared via mucociliary transit. Extending gene vector residence time within the conducting airways is important, but hard to achieve. Gene vector conjugated magnetic particles that can be guided to the conducting airway surfaces could improve regional targeting. Due to the challenges of in-vivo visualisation, the behaviour of such small magnetic particles on the airway surface in the presence of an applied magnetic field is poorly understood. The aim of this study was to use synchrotron imaging to visualise the in-vivo motion of a range of magnetic particles in the trachea of anaesthetised rats to examine the dynamics and patterns of individual and bulk particle behaviour in-vivo. We also then assessed whether lentiviral-magnetic particle delivery in the presence of a magnetic field increases transduction efficiency in the rat trachea. Synchrotron X-ray imaging revealed the behaviour of magnetic particles in stationary and moving magnetic fields, both in-vitro and in-vivo. Particles could not easily be dragged along the live airway surface with the magnet, but during delivery deposition was focussed within the field of view where the magnetic field was the strongest. Transduction efficiency was also improved six-fold when the lentiviral-magnetic particles were delivered in the presence of a magnetic field. Together these results show that lentiviral-magnetic particles and magnetic fields may be a valuable approach for improving gene vector targeting and increasing transduction levels in the conducting airways in-vivo.

show abstract

Section: Methodsmentioning

confidence: 99%

Improved in-vivo airway gene transfer via magnetic-guidance, with protocol development informed by synchrotron imaging

Donnelley

Cmielewski

Morgan

et al. 2022

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Challenges and Barriersmentioning

confidence: 99%

“…Physical barriers that prevent material reaching the airway epithelial cells include the airway surface liquid and glycocalyx in combination with mucociliary clearance 8 , 9 . In many cases the receptors used by viral vectors are not exposed at the apical surface, but instead reside at the basolateral surface obscured by tight junctions 10 . Strategies to promote accessibility to receptors on the basolateral surface have included forms of chemical preconditioning that transiently open the tight junctions 10 .…”

Section: Challenges and Barriersmentioning

confidence: 99%

“…In many cases the receptors used by viral vectors are not exposed at the apical surface, but instead reside at the basolateral surface obscured by tight junctions 10 . Strategies to promote accessibility to receptors on the basolateral surface have included forms of chemical preconditioning that transiently open the tight junctions 10 . There is also the additional barrier of viscous mucus in the airways of CF lungs which is not only difficult to clear from the airways but can also trap inhaled particles and inhibit gene transfer 11 .…”

Section: Challenges and Barriersmentioning

confidence: 99%

See 1 more Smart Citation

Progress in Respiratory Gene Therapy

et al. 2022

View full text Add to dashboard Cite

The prospect of gene therapy for inherited and acquired respiratory disease has energized the research community since the 1980s, with cystic fibrosis, as a monogenic disorder, driving early efforts to develop effective strategies. The fact that there are still no approved gene therapy products for the lung, despite many early phase clinical trials, illustrates the scale of the challenge: in the 1990s, first generation non-viral and viral vector systems demonstrated proof-of-concept but low efficacy. Since then, there has been steady progress towards improved vectors with the capacity to overcome at least some of the formidable barriers presented by the lung. In addition, the inclusion of features such as codon optimisation and promoters providing long-term expression have improved the expression characteristics of therapeutic transgenes. Early approaches were based on gene addition, where a new DNA copy of a gene is introduced to complement a genetic mutation: however, the advent of RNA-based products that can directly express a therapeutic protein or manipulate gene expression, together with the expanding range of tools for gene editing, has stimulated the development of alternative approaches. This review discusses the range of vector systems being evaluated for lung delivery; the variety of cargoes they deliver, including DNA, antisense oligonucleotides, mRNA, siRNA and peptide nucleic acids; and exemplifies progress in selected respiratory disease indications. INTRODUCTION.Respiratory diseases are amongst the leading causes of death globally with numbers rising significantly since 1990 1 . Few effective treatments exist for many indications, often as a result of poor understanding of the disease aetiology. This unmet need has stimulated efforts to develop gene therapeutic approaches for a wide range of genetic, acquired and infectious diseases of the lung. The current era of genomics and transcriptomics has identified causative genes for monogenetic diseases, and a growing understanding of genetic pathways associated with polygenic or acquired conditions. As a result, new potential targets for therapeutic intervention are constantly emerging. This, together with an increasing understanding of stem cells, the availability of induced pluripotent stem (iPS) cells, the ability to direct their differentiation to specific lineages and the rapidly expanding toolbox available for gene replacement/editing is fuelling renewed efforts to develop new gene and cell therapy-based approaches.

show abstract

“…More often, they involve cell types that are present in the airway epithelium (including chronic obstructive pulmonary disease, asthma, and cystic fibrosis) or the alveolar epithelium (such as pneumonia, lung adenocarcinomas, acute lung injury, and idiopathic pulmonary fibrosis) − (Figure ). Because many of these diseases occur as the result of inherited anomalies, RNA-carrying LNPs provide the unique opportunity to use techniques like gene editing, silencing, or replacement therapies to address the root cause of disease rather than just managing symptoms . Because directing LNPs into the lung epithelium via IV injection has proven to be difficult in the past, the administration of LNPs via inhalation has often been explored as a more direct alternative route of delivery. , This introduces the benefits of avoiding rapid hepatic clearance, minimizing delivery to other off-target organs and tissues, and bypassing the need for LNPs to cross the pulmonary endothelial barrier.…”

mentioning

confidence: 99%