Detailed Protocol for the Novel and Scalable Viral Vector Upstream Process for AAV Gene Therapy Manufacturing

Selvaraj, Nagarathinam; Wang, Chao-Kuei; Bowser, Brian; Broadt, Trevor; Shaban, Samir; Burns, Jenna; Saptharishi, Nirmala; Pecháň, Peter; Golebiowski, Diane; Alimardanov, Asaf; Yang, N. Nora; Mitra, Gautam; Vepachedu, Ramarao

doi:10.1089/hum.2020.054

Cited by 19 publications

(19 citation statements)

References 54 publications

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“…A representative HSV system design which utilizes two vectors is shown here, 72 but there is also a similar HSV approach involving just one vector. 75 free media which decreases the risk of adventitious agents, the inability of most human pathogens to replicate in cultured insect cells, and the inability of BVs themselves to replicate in humans. 64,70 Despite the intriguing advantages of BV-based AAV production methods, the manufacturing of AAVs remains generally challenging.…”

Section: Acs Synthetic Biologymentioning

confidence: 99%

Synthetic Biology Design as a Paradigm Shift toward Manufacturing Affordable Adeno-Associated Virus Gene Therapies

2023

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Gene therapy has demonstrated enormous potential for changing how we combat disease. By directly engineering the genetic composition of cells, it provides a broad range of options for improving human health. Adeno-associated viruses (AAVs) represent a leading gene therapy vector and are expected to address a wide range of conditions in the coming decade. Three AAV therapies have already been approved by the FDA to treat Leber’s congenital amaurosis, spinal muscular atrophy, and hemophilia B. Yet these therapies cost around $850,000, $2,100,000, and $3,500,000, respectively. Such prices limit the broad applicability of AAV gene therapy and make it inaccessible to most patients. Much of this problem arises from the high manufacturing costs of AAVs. At the same time, the field of synthetic biology has grown rapidly and has displayed a special aptitude for addressing biomanufacturing problems. Here, we discuss emerging efforts to apply synthetic biology design to decrease the price of AAV production, and we propose that such efforts could play a major role in making gene therapy much more widely accessible.

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Section: Acs Synthetic Biologymentioning

confidence: 99%

Synthetic Biology Design as a Paradigm Shift toward Manufacturing Affordable Adeno-Associated Virus Gene Therapies

2023

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“…AAV vectors are episomal, have excellent safety profiles, and persist long-term in mitotically quiescent cells. Additionally, AAV can be produced to high titers and has a scalability to produce clinical grade vector (Selvaraj et al, 2021). However, the ∼4.7 kb packaging capacity limit is challenging for large transgenes.…”

Section: Gene Therapy Approach: Adeno-associated Virus Vectorsmentioning

confidence: 99%

Gene Therapy Potential for Genetic Disorders of Surfactant Dysfunction

et al. 2022

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Pulmonary surfactant is critically important to prevent atelectasis by lowering the surface tension of the alveolar lining liquid. While respiratory distress syndrome (RDS) is common in premature infants, severe RDS in term and late preterm infants suggests an underlying genetic etiology. Pathogenic variants in the genes encoding key components of pulmonary surfactant including surfactant protein B (SP-B, SFTPB gene), surfactant protein C (SP-C, SFTPC gene), and the ATP-Binding Cassette transporter A3 (ABCA3, ABCA3 gene) result in severe neonatal RDS or childhood interstitial lung disease (chILD). These proteins play essential roles in pulmonary surfactant biogenesis and are expressed in alveolar epithelial type II cells (AEC2), the progenitor cell of the alveolar epithelium. SP-B deficiency most commonly presents in the neonatal period with severe RDS and requires lung transplantation for survival. SFTPC mutations act in an autosomal dominant fashion and more commonly presents with chILD or idiopathic pulmonary fibrosis than neonatal RDS. ABCA3 deficiency often presents as neonatal RDS or chILD. Gene therapy is a promising option to treat monogenic lung diseases. Successes and challenges in developing gene therapies for genetic disorders of surfactant dysfunction include viral vector design and tropism for target cell types. In this review, we explore adeno-associated virus (AAV), lentiviral, and adenoviral (Ad)-based vectors as delivery vehicles. Both gene addition and gene editing strategies are compared to best design treatments for lung diseases resulting from pathogenic variants in the SFTPB, SFTPC, and ABCA3 genes.

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“…The majority of vectors are viral vectors such as adenoviruses ( Zhu et al, 2020 ; Voysey et al, 2021 ; Watanabe et al, 2021 ), retroviruses ( Rastegar et al, 2009 ), lentiviruses ( Maude et al, 2014 ; Sessa et al, 2016 ) or adeno-associated viruses ( Wuh-Liang et al, 2012 ; Keiser et al, 2016 ; Mendell et al, 2017 ; Wang et al, 2018 ). Currently, many scientific groups propose generally safe viral vectors as efficient gene delivery platforms ( Walther and Stein, 2000 ; Bulcha et al, 2021 ; Selvaraj et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Lipid-based nanoparticles and RNA as innovative neuro-therapeutics

et al. 2022

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RNA-delivery is a promising tool to develop therapies for difficult to treat diseases such as neurological disorders, by silencing pathological genes or expressing therapeutic proteins. However, in many cases RNA delivery requires a vesicle that could effectively protect the molecule from bio-degradation, bypass barriers i.e., the blood brain barrier, transfer it to a targeted tissue and efficiently release the RNA inside the cells. Many vesicles such as viral vectors, and polymeric nanoparticles have been mentioned in literature. In this review, we focus in the discussion of lipid-based advanced RNA-delivery platforms. Liposomes and lipoplexes, solid lipid nanoparticles and lipid nanoparticles are the main categories of lipidic platforms for RNA-delivery to the central nervous systems (CNS). A variety of surface particles’ modifications and routes of administration have been studied to target CNS providing encouraging results in vivo. It is concluded that lipid-based nanoplatforms will play a key role in the development of RNA neuro-therapies.

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Detailed Protocol for the Novel and Scalable Viral Vector Upstream Process for AAV Gene Therapy Manufacturing

Cited by 19 publications

References 54 publications

Synthetic Biology Design as a Paradigm Shift toward Manufacturing Affordable Adeno-Associated Virus Gene Therapies

Synthetic Biology Design as a Paradigm Shift toward Manufacturing Affordable Adeno-Associated Virus Gene Therapies

Gene Therapy Potential for Genetic Disorders of Surfactant Dysfunction

Lipid-based nanoparticles and RNA as innovative neuro-therapeutics

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