GENE THERAPY FOR CYSTIC FIBROSIS

Mundappilliyel Anu Joshi, Department of biotechnology, Pillai College of Arts, Commerce

and Science (Autonomous), Panvel, Navi Mumbai, Maharashtra.

Abstract

Cystic fibrosis is an autosomal recessive disease caused by mutations in the cystic fibrosis
transmembrane conductance regulator (CFTR) gene. Gene therapy holds promise for the
treatment of a range of inherited diseases, such as cystic fibrosis. For gene therapy to be
effective in patients with cystic fibrosis, the cDNA encoding the cystic fibrosis
transmembrane conductance regulator protein must be delivered effectively to the nucleus of
the epithelial cells lining the bronchial tree within the lung. The expression of transgene
should be maintained for life time, but it can only maintain for the 30 days for the best.
Various delivery agents are used for this but viral agents can’t be used frequently for the
delivery. The earliest clinical trials in CF patients were conducted in 1993 and used viral and
non-viral gene transfer agents in both the nasal and bronchial airway epithelium. This review
helps to understand the vectors used and the progress in the field of using gene therapy as the
treatment for cystic fibrosis.

Keywords – CFTR, gene therapy, viral vectors, non-viral vectors.

Introduction

Cystic fibrosis (CF)is an autosomal recessive genetic disease affecting > 70,000 individuals
worldwide. CF is caused by mutations in the cystic fibrosis transmembrane conductance
regulator gene (CFTR), which encodes a chloride channel located at the apical membrane of
epithelial cells [1]. Close to 2,000 mutations in this gene have so far been described (see the
Cystic Fibrosis Mutation Database), although fewer than 150 are known to be disease causing
(see the Clinical and Functional Translation of CFTR (CFTR2) website. The cystic fibrosis
phenotype is characterized by progressive lung disease, exocrine pancreatic insufficiency that
results in gastrointestinal malabsorption, intestinal abnormalities that result in malnutrition,
impaired growth, and a variety of other manifestations, including sinusitis and diabetes [2].
Cystic fibrosis is an ideal candidate for gene therapy because (1) it is a single gene defect (2)
it is a recessive condition, with heterozygotes being phenotypically normal (suggesting gene
dosage effects are not critical); (3) the main pathology is in the lung, which is accessible for
treatment; and (4) it is a progressive disease with a virtually normal phenotype at birth,
offering a therapeutic window [3].

Gene therapy in cystic fibrosis

The CFTR gene, mutations of which result in CF, was discovered in 1989; since then
multiple attempts have been made, in the laboratory, preclinical trial, and clinical trial
settings, to correct the underlying defect with gene therapy [3]. For the delivery of cDNA
encoding the cystic fibrosis transmembrane conductance regulator protein to epithelial cells
vectors are required. The ideal vector system would have the following characteristics: (1) an
adequate carrying capacity; (2) to be undetectable by the immune system; (3) to be non-
inflammatory; (4) to be safe to the patients with pre-existing lung inflammation; (5) to have
an efficiency sufficient to correct the cystic fibrosis phenotype; and (6) to have long duration
of expression and/or the ability to be safely re-administered [4]. A variety of different vectors
and delivery techniques have been applied in gene therapy trials. Although nonviral
approaches are becoming increasingly common, viral vectors remain by far the most popular
approach, having been used in approximately two‐thirds of the trials performed to date [5].
To date, more than 20 clinical trials have been conducted with a variety of viral and nonviral
gene transfer agents [3].

Viral vectors

Gene delivery systems have been derived from various viruses. Adenoviral, adeno-associated
viral, lentiviral vectors, poxvirus, Sendai and herpes virus are investigated for gene transfer.
In general, the main advantage of viral vectors is the high transduction efficiency in vivo,
compared to current synthetic systems. The first viral GDS for the treatment of CF that was
tested in primates and clinical trial was the adenoviral vector [6]. There are other viral vector
systems under development, the most promising being pseudo typed lentiviral vectors and
parainfluenza virus type 3. Lentiviral vectors offer the possibility of providing sustained
expression of the therapeutic gene, as they integrate their genome into the host DNA [4].

Non-viral vectors

Non-viral vectors have the potential to avoid some of the critical problems observed with
viral vectors, such as the immune response, limited packaging capacity, and random
integration. There are three main non-viral vector systems: cationic liposomes, DNA–
polymer conjugates and naked DNA. To date, only cationic liposome-based systems have to
circumvent lysosomal degradation, though much less efficient than virus derived GDS [6].
Clinical trials have demonstrated that CFTR gene transfer to the airway can be achieved
safely and, for nonviral vectors, repeatedly. However, side effects were observed with both
viral and nonviral approaches and, in general, the duration of expression was relatively short
[3].

Conclusion

During years of clinical research in CF gene therapy, researchers have proved the principle of
gene delivery to the CF airway and correction of some of the basic functions of the CFTR
protein [3]. For monogenetic hereditary diseases, gene therapy offers the potential of
correcting the underlying cause for which the responsible gene is known. Since delivery of a
CFTR gene in the relevant cells, at the right time, and at the proper expression level seems
difficult to achieve with available vector systems, alternatives should be considered [6].

References

[1] Josef Rosenecker, Stephanie Huth & Carsten Rudolph, Gene therapy for cystic fibrosis
lung disease: Current status and future perspective.

[2] Felix Ratjen, Scott C. Bell, Steven M. Rowe, Christopher H. Goss,

Alexandra L. Quittner and Andrew Bush, Cystic fibrosis.

[3] Jane C. Davies and Eric W. F. W. Alton, Gene Therapy for Cystic Fibrosis, Department
of Gene Therapy, Imperial College London, London, United Kingdom.

[4] Tim W. R. Lee, David A. Matthews and G. Eric Blair, Novel molecular approaches to
cystic fibrosis gene therapy.

[5] Samantha L. Ginn, Anais K. Amaya, Ian E. Alexander, Michael Edelstein, Mohammad R.
Abedi, Gene therapy clinical trials worldwide to 2017.

[6] Daniel Klinka, Dirk Schindelhauerb, Andreas Lanerc, Torry Tuckerd, Zsuzsanna Bebokd,
Erik M. Schwiebertd, A. Christopher Boyde, Bob J. Scholte, Gene delivery systems—gene
therapy vectors for cystic fibrosis.