Advancing CGT: A GENEWIZ & Brooks Life Sciences Virtual Event

The adeno-associated virus (AAV) has evolved overlapping genes to maximize its genome use, as with the recently-discovered ORF in the cap gene which encodes a membrane-associated accessory protein (MAAP) located in the same genomic region as the VP1/2 unique domain. This 13 KDa protein, unique to the dependovirus genus, is not homologous to any known protein. To study the role of MAAP in wild-type AAV (wt-AAV), we made point mutations along the MAAP ORF while keeping overlapping capsid proteins' ORFs intact. In cells co-transfected with plasmids encoding wt-AAV genome and adenovirus helper genes, MAAP localized in the plasma membrane, as well as intracellular membranes. Both inactivation and truncation of MAAP translation affected the emergence and intracellular distribution of the AAV capsid proteins. Importantly, while MAAP was beneficial for wt-AAV replication/infection, it was not essential for AAV generation. Oppositely, some MAAP-modifications resulted in substantially improved virus yields and capsid integrity. Altogether, our results reveal some MAAP functions having important implications for better-quality and quantity production of AAV vectors for therapeutic purposes.

Advanced therapy medicinal products (ATMPs) are ‘living drugs’ that have proven highly effective at treating diseases, such as cancer, that were once thought untreatable. As ‘living drugs’, ATMPs require complex chemistry, manufacturing, and controls (CMC) to enable efficient commercialization and maintain therapeutic consistency and potency. Part of these controls is the need to implement optimizing freezing protocols to facilitate the long-term frozen storage and transport of ATMPs from the manufacturing site to the clinic. Cryopreservation for ATMPs requires specialized storage conditions and distribution channels to maintain ATMPs at ultralow temperatures (e.g. below -80°C) that permit proper biological function post-thaw. As evidenced by the recent COVID vaccine approvals, cryogenic storage and distribution at ultralow temperatures is a challenge for the existing pharmaceutical infrastructure and a hinderance to the widespread distribution of ATMPs. Solutions to overcome the challenges of cell cryopreservation, storage, and distribution at ultralow temperatures need to be identified, validated, and implemented to increase ATMP adoption for the general population. This presentation will introduce the audience to the challenges associated with ATMP cryopreservation, as well as provide potential solutions for how these challenges can be tackled during ATMP development, from pre-clinical through commercialization.

Cell therapies, including chimeric antigen receptor (CAR) T-cell therapy, are emerging as promising treatments, especially for hematological malignancies. However, many challenges remain in developing this emerging class of therapeutics, from deepening responses in blood-based tumors to expanding efficacy in solid tumors. To face these challenges, understanding the mechanisms behind response, resistance, and toxicity to cell therapies is key. Tools with the resolution and scalability to reveal intra- and inter-patient heterogeneity across cancer types will enable researchers to identify key biomarkers and optimize therapeutic products. Explore how 10x Genomics single-cell solutions can help uncover molecular insights into cell therapeutic efficacy and toxicity, transforming the way we approach immune cell therapies for cancer.

Interest in cell and gene therapy-based disease prevention and treatment has increased rapidly over the last few decades, however there are still many hurdles to overcome and further progress to be made in the field. In this presentation, Dr. Elizabeth Louie from Brooks Life Sciences will discuss our innovative cell and gene therapy solutions across the clinical development pipeline. She will highlight GENEWIZ’s adeno-associated virus (AAV) plasmid DNA sequencing (RUO/GLP), preparation, and synthesis solutions for recombinant AAV (rAAV) production, which includes confirmation, maintenance, and correction of inverted terminal repeat (ITR) regions. She will also talk about GENEWIZ’s proprietary Sanger & NGS viral genome sequencing confirmation solutions post-AAV virus packaging, including at the regulatory level, to optimize AAV production for gene therapy development. Finally, Dr. Louie will spotlight Brooks Life Sciences’ robust cold chain infrastructure solutions and expertise, demonstrating how having the appropriate systems and processes in place to maintain strict cold chain-of-custody, while getting the right treatment to the right patient at the right time, addresses this challenge in the industry. A brief overview of biomaterial storage and transport solutions will be capped off with a mention of cold chain products and field service offerings to support research from benchtop to bedside.