ABSTRACT
Background & Aim: The recent supply chain crisis highlights a need to establish alternative manufacturing (MFG) protocols ensuring continuity of existing and new cell therapy (CT) clinical trials. Our academic CT program, and likely others, experienced purchasing delays and restrictions caused by diversion of critical supplies to meet COVID-19- related research demands and/or reduced vendor capacity due to resource constraints, including attrition of skilled workforce. Mitigation strategies aimed at creating process redundancies overcome production challenges resulting from a scarcity of goods. Here, we validated an alternative ex vivo culture system to clinically MFG lentiviral vector (LV) modified CAR T cells due to limited availability of cell expansion culture bags for the Wave bioreactor, a critical unit of operation that we have used to successfully MFG thousands of gene-modified T cell products for 30+ clinical trials. Methods, Results & Conclusion: The disposable G-REX culture vessels were compatible and seamlessly integrated with our closed system platform. Mesothelin CAR T cells were manufactured in parallel via the G-REX or conventional Wave bioreactor using consented patient starting material. Critical quality attributes of the final T cell products, including viability, transduction efficiency, phenotype and function were assessed. Transduction efficiencies assessed by flow cytometry and/or molecular qPCR were lower in products generated in the G-REX compared to the wave using the same multiplicity of infection. However, at least 50-fold expansion was achieved, with cell viabilities greater than 90% and with comparable cellular phenotypes. The Meso CAR T cells generated by either process were capable of eliciting CAR-mediated cytotoxicity and effector cytokine production. Strikingly, 2-4 billion T cells were harvested from a starting seed number of just 50 million T cells in the 1L G-REX, which may be sufficient to meet most protocol- specified cell therapy doses, suggesting that a full apheresis collection may not be needed. Notably, this process required just 1/3 of the starting material, 1/5 of the media and decreased manual effort through culture duration compared to the Wave. Additionally, the reduced reliance on specialized capital equipment combined with a small footprint enables simultaneous MFG of several immunotherapy products. These advantages propose consideration in replacement of current expansion platform as well as validating an alternative process for MFG CAR T cells.
ABSTRACT
Objectives: The COVID-19 pandemic has led to immense challenges for healthcare systems worldwide. People with cystic fibrosis (CF) were included in the clinically extremely vulnerable group for complications of coronavirus by the UK government and advised to shield during a national lockdown. Data suggests that pandemic-related restrictions have been linked to a reduction in pulmonary exacerbation events (PEx). We sought to explore whether an increase in medicine possession ratio and potentially adherence may be a factor in this finding. Methods: 50 patients who received medication through a homecare delivery system at a single large adult centre were randomly selected. Data from 12 months ‘pre-lockdown’ was compared to data for 9 months following start of shielding in March 2020. MPR was calculated and capped at 100%. Medications that were started or stopped during the pandemic were not included. Wilcoxon signed rank test was used to compare pre- and post-values. Results: 91 prescription medications were valid for analysis (45 nebulised antibiotics, 34 mucolytics and 12 CFTR modulators). MPR increased for 41 prescriptions (45.1%), decreased for 21 medications (23.1%) and remained unchanged for 29 medications (31.9%). Median MPR increased from 83% [57–100%] to 89% [66–100%], p = 0.037. MPR for nebulised antibiotics significantly increased (median 75% [54–100%] vs 89% [61–100%], p = 0.027). Median MPR for CFTR modulators was 100% throughout and did not change for mucolytics (75% [42–100%] vs 78% [53–100%], p = 0.419). Conclusion: We report a significant change in medication possession in adults with CF during the coronavirus pandemic in the UK. It is unclear whether this change translated to an increase in adherence but may be one factor in the reported decrease in PEx events described during this time. It is notable that increases were largely driven by inhaled antibiotics and this may represent a concerted effort to achieve maximal protection from infection.