Innovative therapies in genetic diseases: Cystic fibrosis

Cystic fibrosis, also named mucoviscidosis, is the most frequent hereditary pulmonary disease and is produced by mutations in the CFTR gene, encoding an anionic channel for chloride and bicarbonate involved in the regulation of salt and bicarbonate metabolisms. Currently, about half of the patients with cystic fibrosis can benefit personalized therapy consisting in modulators, drugs which restore or improve the functionality and stability of CFTR. Moreover, presently, other therapies, such as gene therapy using the CRISP/CAS-9, modified antisense oligonucleotides or the insertion of the wild-type gene using nanolipidic particles or viral vectors, are being developed. This article aims to take stock of the principal types of cystic fibrosis therapies which have been approved or are in clinical trials.

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Potential of helper-dependent Adenoviral vectors in CRISPR-cas9-mediated lung gene therapy

Cell & Bioscience ◽

10.1186/s13578-021-00662-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ranmal Avinash Bandara ◽

Ziyan Rachel Chen ◽

Jim Hu

Keyword(s):

Cystic Fibrosis ◽

Gene Therapy ◽

Lung Disease ◽

Viral Vectors ◽

Gene Editing ◽

Genetic Diseases ◽

Adenoviral Vectors ◽

The Status

AbstractSince CRISPR/Cas9 was harnessed to edit DNA, the field of gene therapy has witnessed great advances in gene editing. New avenues were created for the treatment of diseases such as Cystic Fibrosis (CF). CF is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. Despite the success of gene editing with the CRISPR/Cas9 in vitro, challenges still exist when using CRISPR/Cas9 in vivo to cure CF lung disease. The delivery of CRISPR/Cas9 into lungs, as well as the difficulty to achieve the efficiency required for clinical efficacy, has brought forth new challenges. Viral and non-viral vectors have been shown to deliver DNA successfully in vivo, but the sustained expression of CFTR was not adequate. Before the introduction of Helper-Dependent Adenoviral vectors (HD-Ad), clinical trials of treating pulmonary genetic diseases with first-generation viral vectors have shown limited efficacy. With the advantages of larger capacity and lower immunogenicity of HD-Ad, together with the versatility of the CRISPR/Cas9 system, delivering CRISPR/Cas9 to the airway with HD-Ad for lung gene therapy shows great potential. In this review, we discuss the status of the application of CRISPR/Cas9 in CF gene therapy, the existing challenges in the field, as well as new hurdles introduced by the presence of CRISPR/Cas9 in the lungs. Through the analysis of these challenges, we present the potential of CRISPR/Cas9-mediated lung gene therapy using HD-Ad vectors with Cystic Fibrosis lung disease as a model of therapy.

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Vectors for Glioblastoma Gene Therapy: Viral & Non-Viral Delivery Strategies

Nanomaterials ◽

10.3390/nano9010105 ◽

2019 ◽

Vol 9 (1) ◽

pp. 105 ◽

Cited By ~ 26

Author(s):

Breanne Caffery ◽

Jeoung Lee ◽

Angela Alexander-Bryant

Keyword(s):

Gene Therapy ◽

Clinical Trials ◽

Glioblastoma Multiforme ◽

Viral Vectors ◽

Survival Rates ◽

Genetic Alterations ◽

Primary Brain Tumor ◽

Promising Alternative ◽

Gene Vectors ◽

Polymeric Vectors

Glioblastoma multiforme is the most common and aggressive primary brain tumor. Even with aggressive treatment including surgical resection, radiation, and chemotherapy, patient outcomes remain poor, with five-year survival rates at only 10%. Barriers to treatment include inefficient drug delivery across the blood brain barrier and development of drug resistance. Because gliomas occur due to sequential acquisition of genetic alterations, gene therapy represents a promising alternative to overcome limitations of conventional therapy. Gene or nucleic acid carriers must be used to deliver these therapies successfully into tumor tissue and have been extensively studied. Viral vectors have been evaluated in clinical trials for glioblastoma gene therapy but have not achieved FDA approval due to issues with viral delivery, inefficient tumor penetration, and limited efficacy. Non-viral vectors have been explored for delivery of glioma gene therapy and have shown promise as gene vectors for glioma treatment in preclinical studies and a few non-polymeric vectors have entered clinical trials. In this review, delivery systems including viral, non-polymeric, and polymeric vectors that have been used in glioblastoma multiforme (GBM) gene therapy are discussed. Additionally, advances in glioblastoma gene therapy using viral and non-polymeric vectors in clinical trials and emerging polymeric vectors for glioma gene therapy are discussed.

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Codon optimization of human factor VIII cDNAs leads to high-level expression

Blood ◽

10.1182/blood-2010-05-282707 ◽

2011 ◽

Vol 117 (3) ◽

pp. 798-807 ◽

Cited By ~ 109

Author(s):

Natalie J. Ward ◽

Suzanne M. K. Buckley ◽

Simon N. Waddington ◽

Thierry VandenDriessche ◽

Marinee K. L. Chuah ◽

...

Keyword(s):

Gene Therapy ◽

Factor Viii ◽

Viral Vectors ◽

Hemophilia A ◽

Lentiviral Vector ◽

Wild Type ◽

Fviii Activity ◽

Human Factor Viii

Abstract Gene therapy for hemophilia A would be facilitated by development of smaller expression cassettes encoding factor VIII (FVIII), which demonstrate improved biosynthesis and/or enhanced biologic properties. B domain deleted (BDD) FVIII retains full procoagulant function and is expressed at higher levels than wild-type FVIII. However, a partial BDD FVIII, leaving an N-terminal 226 amino acid stretch (N6), increases in vitro secretion of FVIII tenfold compared with BDD-FVIII. In this study, we tested various BDD constructs in the context of either wild-type or codon-optimized cDNA sequences expressed under control of the strong, ubiquitous Spleen Focus Forming Virus promoter within a self-inactivating HIV-based lentiviral vector. Transduced 293T cells in vitro demonstrated detectable FVIII activity. Hemophilic mice treated with lentiviral vectors showed expression of FVIII activity and phenotypic correction sustained over 250 days. Importantly, codon-optimized constructs achieved an unprecedented 29- to 44-fold increase in expression, yielding more than 200% normal human FVIII levels. Addition of B domain sequences to BDD-FVIII did not significantly increase in vivo expression. These significant findings demonstrate that shorter FVIII constructs that can be more easily accommodated in viral vectors can result in increased therapeutic efficacy and may deliver effective gene therapy for hemophilia A.

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Mmp10 is required for post-translational methylation of arginine at the active site of methyl-coenzyme M reductase

10.1101/211441 ◽

2017 ◽

Cited By ~ 3

Author(s):

Zhe Lyu ◽

Chau-wen Chou ◽

Hao Shi ◽

Ricky Patel ◽

Evert C. Duin ◽

...

Keyword(s):

Active Site ◽

Methane Formation ◽

Wild Type ◽

Coenzyme M ◽

Methane Cycle ◽

Gene Encoding ◽

Post Translational Modifications ◽

Whole Cells ◽

Type Gene ◽

Methyl Coenzyme M Reductase

AbstractCatalyzing the key step for anaerobic methane production and oxidation, methyl-coenzyme M reductase or Mcr plays a key role in the global methane cycle. The McrA subunit possesses up to five post-translational modifications (PTM) at its active site. Bioinformatic analyses had previously suggested that methanogenesis marker protein 10 (Mmp10) could play an important role in methanogenesis. To examine its role, MMP1554, the gene encoding Mmp10 inMethanococcus maripaludis, was deleted with a new genetic tool, resulting in the specific loss of the 5-(S)-methylarginine PTM of residue 275 in the McrA subunit and a 40~60 % reduction in the maximal rates of methane formation by whole cells. Methylation was restored by complementations with the wild-type gene. However, the rates of methane formation of the complemented strains were not always restored to the wild type level. This study demonstrates the importance of Mmp10 and the methyl-Arg PTM on Mcr activity.

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Progress Toward The Synthesis Of Labeled Derivatives Of The Cystic Fibrosis Drug Ivacaftor

10.32920/ryerson.14665104.v1 ◽

2021 ◽

Author(s):

Salma Elmallah

Keyword(s):

Cystic Fibrosis ◽

Plasma Membrane ◽

Genetic Diseases ◽

Early Death ◽

Open Probability ◽

Gene Encoding ◽

Short Period ◽

Cftr Mutations ◽

Cftr Protein ◽

Orally Bioavailable

Cystic fibrosis (CF) is one of the most common genetic diseases, affecting approximately 70,000 people worldwide causing severe complications and often leading to early death. CF is caused by a mutation in the gene encoding for the cystic fibrosis transmembrane conductance regulator (CFTR) protein which is responsible for fluid and ion transport through epithelial membranes maintaining the formation of a thin slippery mucous layer. CFTR mutations either lead to a trafficking defect where the CFTR protein does not reach the plasma membrane or a gating defect where CFTR protein at the plasma membrane does not function properly. Treatment of cystic fibrosis usually addresses the symptoms to overcome the complications of the disease such as pneumonia, lung infections, pancreatitis, maldigestion and infertility. Vertex pharmaceuticals has been interested in developing small molecules that have the ability to interact with mutated CFTR proteins, aiding in their delivery to the cell membrane and/or restoring their channel function. VX-770 is an orally bioavailable potentiator that has the ability to improve the gating activity and increasing the open probability of CFTR protein in patients carrying the G551D mutation. VX770, Ivacaftor, was recently approved by the US FDA after showing very good improvements in the lung function in CF patients with good safety profile. Our research is focusing on the synthesis of VX770 under mild conditions and formation of labeled derivatives to help in the understanding of its exact mode of action. Different methods were developed toward the synthesis of the two main components, LHS and RHS, of VX770 by using less harsh conditions for a short period of time. We were successfully able to make two photoaffinity labeled derivatives, aryl azide and benzophenone derivatives, which will be beneficial in tracking the drug molecule and revealing the exact site of interaction between the drug and the protein. Synthesis of VX770 fragments was is another focus of interest in our research as they will provide us with information about the best positions for further derivatization.

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Not as easy as it looks: Gene therapy for cystic fibrosis

The Biochemist ◽

10.1042/bio03003022 ◽

2008 ◽

Vol 30 (3) ◽

pp. 22-25

Author(s):

Tim Lee ◽

Eric Blair

Keyword(s):

Cystic Fibrosis ◽

Gene Therapy ◽

Clinical Trials ◽

Respiratory Tract ◽

Low Levels

The gene responsible for cystic fibrosis (CF) was first identified in 1989. Since that time, gene therapy has held the promise of a cure for cystic fibrosis; however, progress has been much slower than initially hoped. Clinical trials of gene therapy have demonstrated that the corrected CF gene can be expressed successfully in the respiratory tract of subjects with CF, but only for approximately 30 days at best, and at low levels.

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TREATMENT POSSIBILITIES FOR ACQUIRED AND HEREDITARY DISEASES BY GENE THERAPY: A REVIEW

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2021v13i2.39995 ◽

2021 ◽

pp. 26-32

Author(s):

P. V. KAMALA KUMARI ◽

G. EKSHITHA, V. HARIKA

Keyword(s):

Gene Therapy ◽

Nucleic Acids ◽

Viral Vectors ◽

Target Gene ◽

Genetic Diseases ◽

Hereditary Diseases ◽

Root Cause ◽

Delivery Strategies ◽

Foreign Genes ◽

And Control

Therapeutic nucleic acids demand specificity and accuracy in design as well as delivery strategies used in replacement or silencing of the target gene. Gene therapy is believed to be the therapy in which the root cause of the diseases can be treated at the molecular level. Generally gene therapy helps in the identification of the origin of the disorder instead of using drugs to diminish or control the symptoms. The application of nucleic acids to treat and control diseases is known as “gene therapy.” Gene therapy consists on the substitution or addition of a functional gene into the nucleus of a living cell, in order to treat a disease or repair a dysfunction, caused by this gene failure. This therapy is used to correct defective genes, which are responsible for genetic diseases. Thus, gene therapy can be used to prevent, treat or regulate hereditary or acquired disorders, by the production of therapeutic proteins. The gene therapy is mediated by the use of viral and non-viral vectors to transport foreign genes into somatic cells to restorative defective genes. This review focuses on viral vectors in detail.

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Next-Generation Gene Therapy for Parkinson’s Disease Using Engineered Viral Vectors

Journal of Parkinson s Disease ◽

10.3233/jpd-212674 ◽

2021 ◽

pp. 1-9

Author(s):

Tomas Björklund ◽

Marcus Davidsson

Keyword(s):

Parkinson’S Disease ◽

Gene Therapy ◽

Parkinson's Disease ◽

Clinical Trials ◽

Genome Editing ◽

Viral Vectors ◽

State Of The Art ◽

The State ◽

Next Generation ◽

Suitable Vector

Recent technological and conceptual advances have resulted in a plethora of exciting novel engineered adeno associated viral (AAV) vector variants. They all have unique characteristics and abilities. This review summarizes the development and their potential in treating Parkinson’s disease (PD). Clinical trials in PD have shown over the last decade that AAV is a safe and suitable vector for gene therapy but that it also is a vehicle that can benefit significantly from improvement in specificity and potency. This review provides a concise collection of the state-of-the-art for synthetic capsids and their utility in PD. We also summarize what therapeutical strategies may become feasible with novel engineered vectors, including genome editing and neuronal rejuvenation.

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