• Coyne Hubbard posted an update a month ago

    The innovative therapy, chimeric antigen receptor (CAR) T-cells, is considered a living drug and presents a compelling alternative to standard anticancer treatments. T-cells are redirected, via gene-editing techniques, to a specific cancer cell surface target antigen, employing a synthetic chimeric antigen receptor (CAR) protein. A modular architecture of cars incorporates four principal elements: an antigen-binding domain, a hinge region, a transmembrane domain, and one or more intracellular signaling domains necessary for T-cell activation. Striking a balance between product quality, industrial-scale production capabilities, and cost-effectiveness in CAR T-cell manufacturing is a substantial challenge, particularly when moving from the confines of clinical trials to commercial availability across many collection, manufacturing, and treatment sites. A significant barrier is maintaining product consistency in the presence of the inherent variability characteristic of autologous materials. To resolve these limitations, a deep understanding of the product and its biological mechanisms of action is paramount to establishing a precisely defined target product profile, including a list of specific quality attributes to be rigorously assessed in every batch before certification. As this area of study advances, new hurdles appear, particularly regarding the safety implications of using allogenic T-cells and gene-editing tools. The chapter will discuss the release and potency assays indispensable for CAR T-cell manufacturing, exploring their significance, present challenges, and future trends.

    Cellular therapy products are evaluated, in part, through potency testing. In vitro, identified quality-related biomarkers are often quantified in a laboratory setting. In spite of these aspects, the relatively unstable nature of the majority of cellular therapy products, the variance between batches, and the restricted time for evaluation currently discourage their widespread implementation. Thanks to the advancement of material technology and miniaturization techniques within the past two decades, researchers have been able to direct their attention to the Lab-on-Chip concept for diagnostic applications. To expand the application range of healthcare services, these devices enable portable, rapid, sensitive, automated, and affordable biochemical analyses. Nevertheless, one could contend that the hopes for their affordability are not fully met, primarily because of the lack of a tangible, workable manufacturing process. Developing economical diagnostic platforms utilizing the Lab-on-Printed Circuit Board (Lab-on-PCB) technique demonstrates substantial potential by capitalizing on the considerable economies of scale in the long-standing PCB sector. The Lab-on-Chip concept benefits from the PCB platform’s apparently singular integration capabilities for electronics and microfluidics. In this chapter, the progress of Lab-on-PCB prototypes is analyzed, quantitatively assessing critical quality attributes within miniature microchips. Potential for use in potency testing is explored. With a focus on their technology and its applications, we will simultaneously examine its potential for practical usage and commercialization within the marketplace.

    Regulatory approval criteria, critical for cell product application, must be met before infusion into patients, following thorough characterization within cell manufacturing facilities. Amongst the leading cell therapy candidates in clinical trials worldwide, mesenchymal stromal cells (MSCs) hold a prominent position. Early-phase clinical trials have revealed that mesenchymal stem cells exhibit a strong safety profile and are well-accepted by the patients. MSCs, despite initial promise, have displayed contrasting effectiveness in later stages of clinical trials. Potential explanations for these discrepancies include the currently incompletely understood mechanisms driving their therapeutic effects. Due to the probable involvement of numerous factors in MSC-derived clinical outcomes, an assay targeting only a single quality might fail to fully represent potency. Consequently, a combination of bioassays and analytical techniques, encompassing an assay matrix, is a more suitable approach for determining MSC potency precisely. Quantitative assessment of MSC attributes, including immunological profiles, genomic characteristics, secretome composition, phosphorylation markers, morphological features, biomaterial interactions, angiogenic potential, and metabolic function, is achieved through the advanced technologies and targets discussed in this chapter.

    In synovial joints, articular cartilage, functioning as a shock absorber, is found at the ends of bones, facilitating their movement. Treatment for damaged articular cartilage is essential, as it does not repair itself and this damage can lead to osteoarthritis. Osteoarthritis is characterized by a progressive degradation of cartilage and inflammation throughout the joint tissues. epz004777 inhibitor Despite its proven efficacy, autologous chondrocyte implantation suffers from the significant impediment of demanding two separate surgical operations. Multipotent mesenchymal stromal cells (MSCs) have demonstrated the ability to facilitate one-step cartilage repair. The potential of mesenchymal stem cells (MSCs) in treating osteoarthritis is attributed to their remarkable immunomodulatory and regenerative properties. In addition, since the paracrine effects of mesenchymal stem cells (MSCs) are partly attributed to extracellular vesicles (EVs), tiny membrane-bound particles released by cells, EVs are now being intensely studied for their potential therapeutic advantages. While mesenchymal stem cells (MSCs) have found their way into clinical cartilage therapies and extracellular vesicles (EVs) are employed in in vivo effectiveness assessments, the determination of their potency and the development of potency assays have received relatively little consideration. This chapter examines considerations and suggestions for the design of potency assays to assess the therapeutic potential of mesenchymal stem cells (MSCs) and their vesicles (MSC-EVs) in managing cartilage defects and osteoarthritis.

    A significant amount of research has been dedicated to examining the potential of cells as advanced therapeutic medicinal products in treating pathologies of the skeletal system. The osteoblast lineage, originating from a rare bone marrow subpopulation of cells that readily adhere to plastic surfaces in culture, is responsible for the production and subsequent mineralization of the extracellular matrix during bone development. Fibroblastoid cells, conveniently forming single-cell derived colonies, termed colony-forming unit fibroblasts, are capable of subsequent differentiation into aggregates resembling small regions of cartilage or bone. Variability in donor characteristics and the loss of osteogenic potential during prolonged cell culture have complicated the search for trustworthy potency assay markers. Despite this, functional osteoblast models, derived from telomerized human bone marrow stromal cells, have permitted in-depth comparative analyses of gene expression, microRNA profiles, morphological phenotypes, and secreted protein profiles. This chapter explores the intricate molecular mechanisms driving osteogenic differentiation in multipotent stromal cells and bone development, examining the selection of quantifiable biomarkers for osteogenic potency assays.

    Cell-based products, significantly altered for human consumption, are classified as medicines, requiring detailed characterization of their identity, purity, and potency by regulatory authorities. Identifying and quantifying the latter crucial quality attribute presents the greatest challenge, demanding potency assays that accurately mirror the intended mechanism of action and demonstrate the drug’s biological impact. Despite this, the operational mechanisms involved are generally not fully understood, making the precise definition and validation of suitable potency tests a significant hurdle, a formidable quest. Although substantial work still remains within the scientific field, this chapter concentrates on the present strategies used by cell and gene therapy developers to showcase the potency of new medicines. It delves into the regulatory environment and the necessity for standardization to clarify critical considerations in designing a potency assay.

    The capacity of an Advanced Therapy Medicinal Product (ATMP) to elicit the required clinical effect, that is, potency, is a quantitative measure of its biological activity. The quality control strategy for ATMP batch release and market application includes potency testing, which is essential for both. In order to ensure accuracy and dependability, a potency assay needs to be developed. For assay development focused on potency, the product’s mode of action must first be established, thereby defining the critical biological activity to be measured. The implementation of a potency assay should be initiated early on in the product development cycle, and it should be progressively adopted by the ATMP’s manufacturing, quality control, and release procedures. Potency testing, with its broad spectrum of applications, is a cornerstone of clinical practice. The potency assay serves as a crucial instrument in evaluating the product’s stability, detecting the consequences of modifications in the manufacturing process, showcasing consistent quality and manufacturing standards across batches, determining clinical efficacy, and establishing the effective dose required. This chapter examines the requirements and difficulties encountered in the development of potency assays, emphasizing the importance of a well-characterized potency assay for clinical applications.

    Potency assays represent indispensable experiments, situated at the hub of the comprehensive intricacy surrounding cellular therapies. In addition, a broad array of considerations surpassing biological and scientific criteria are critical to producing validated potency assays that satisfy regulatory agency acceptance criteria for innovative advanced therapy medicinal products. Though periods of experimentation and advancement in cell therapy can be frustratingly lengthy, progress is presently happening at a remarkably accelerated pace, aided by potency assays that emphasize the need for a thorough analysis of the key elements driving the therapeutic mechanism.

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