CGT 4.0: a distant dream or inevitable future? Smart process automation is critical to make efficient scalability of CGT manufacturing a reality

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: March 19, 2025

1IntroductionCell and gene therapies (CGTs) are new treatment modalities with demonstrated clinical results against a wide range of hard-to-treat diseases.[1, 2] They either ex vivo treatments, obtained by manipulating cells in laboratory before returning them to the patient, or applications, that involve direct injection genetic material into bloodstream target organ.[3] Cell therapy (CGT) products significantly different from previous generations biologics, such as recombinant proteins vaccines, have been challenging production capabilities, supply chain business models pharmaceutical industry.[4, 5] With CGT market still its infancy, even decade after first approvals, three factors related their manufacturing limiting broad adoption clinic: 1) highly variable starting materials due personalised nature treatments[6, 7], 2) diversity complexity processes[8], 3) lack fit-for-purpose tools supporting scalable at commercial stage[9]. The combination these leads limited availability high cost CGTs, making it reach success while equitable patient access.[3, 10]To tackle this challenge, authors article partnered PAT4CGT consortium aims develop miniaturised process analytical technology (PAT) platform, tailored specifically for manufacturing. Advanced PAT is critical understanding R&D also becoming key tool monitoring during manufacturing.[11, 12] This trend aligned concept Industry 4.0, transform systems through integration advanced digital technologies.[13] Adoption 4.0 principles paradigm shift, introducing cutting-edge, technologies, moving away centralised, manual paper-led, towards distributed, automated knowledge-driven processes.[14] In our opinion, solutions innovative approaches support essential deliver on promise novel modalities. Thus, goal project lay foundation minimally-invasive parameters (CPPs) CGT. To end, we standalone, closed, automated, sensor platform line CPPs, suited applications manufacturing.2Closed, single-use cell culture processingWe believe layers innovation necessary achieve transformation (Figure 1). fundamental layer consists perform two unit operations core all bioprocesses: culture[15] processing[16]. These steps traditionally performed bioreactors centrifugation systems, respectively. Single-use closed become standard small- mid-scale biologics increasingly used large scale processes.[17] While there technologies implemented large-scale upstream processes, remain hard scale. Overall, need approaches[18] take consideration unique properties set apart biopharmaceutical products.[19, 20]In case autologous, gene-edited therapies, needed complex, multi-step processes streamlined sequence operations. Replacing automation systems[21, 22] has potential increase standardisation, robustness, scalability reproducibility lowering labour clean room infrastructure costs.[23-25] There empirical evidence[26] automation[27] can substantially reduce "hands-on" operator time required CAR T-cell therapies.[28] Similarly, higher transduction lower variability between batches were haemopoietic stem using semi-automated approach, when compared process.[29] Novel enabling end-to-end autologous will be field enable point care production. modular being implemented, bringing single device, thus removing transfer tools, lead yields product quality. Solutions manipulations across volumes numbers instrumental unlocking full treatments. Furthermore, dose within same device costs currently invested transition pre-clinical stage.In therapy, cryopreservation thawing master bank step process. methods allow handling banks use (high density) bag-based approach microfluidic-based system. Immortalised lines (e.g. HEK-derived cells) already characterised exist viral vector manufacturing.[30] However, industry lacking fully workflows covering steps, expansion, virus purification up final formulation.[31] traditional operations, flatware flask ultracentrifugation purification, unsatisfactory they not only hinder but possibility continuous data acquisition. 3Miniaturised sensors monitoringThe middle composed measure CPPs real-time, providing vital information about status. Monitoring capabilities capture record development batch documentation provide inputs control.[32] facilitates decision early detection deviations out specification (OOS) events. timely intervention improve rate, well minimisation losses associated failure. this, variety offline, inline remains questionable.[12] concurring approaches: (i) measuring sample high-precision method HPLC-MS) automating sampling process, (ii) relying an situ spectroscopy) often indirect measurement principle requires extensive analysis applied system without risk contamination. arrays biologics' – temperature, pH, dissolved oxygen CO2 insufficient characterise status complex because tracking metabolism development.[33-36] historical design medium supplementation strategy, absence least frequent limits flexibility adjust media components important changes occur, hindering productivity Inline spectroscopic sensors, infrared, Raman dielectric spectroscopy, offer monitor, nutrients, metabolites[37-40] count.[41, 42] influenced non-specific (background) variations.[41] Moreover, interpretation, validation software control, physical probe innovative, small-scale CGTs challenging.[41]The limitations background signals above-mentioned overcome specific recognition element, antibody-based enzymatic biosensor.[43, 44] These, however, require representative sample. technique must requirements production, mainly scales. Sterile opens further downstream processing offline analytics. general, collection samples environment precise, contamination continuous/frequent measurements. play role increasing subsequent analysis. 4Digital twin knowledge controlThis us top which analytics solution allowing information, collected actionable knowledge, efficiently manage lifecycle. mathematical model known "digital twin", integration, aim predictive control quality attributes (CQAs) adjusting CPPs.[45] Historically, experimental mechanistic modelling balance substrates metabolites) describe microbial systems.[46] successfully adapted more cases, Chinese hamster ovary (CHO) cells.[47, 48] assumptions universal transferred types.[49] imminent translation generally low CGTs.[50-52] Therefore, prerequisite developing accurate clearly define, then time-resolved values CQAs processes,[53] in-situ central role.In view, purely data-driven, neural network could processes.[54, 55] For generation amounts train models, novel, culturing resolve bottleneck current capacities. Another apply sophisticated algorithms, hybrid modelling. Hybrid combines representations data-driven methods.[56] instance, long-short term memory[57] physics informed approaches,[58] reinforced learning-based methods.[59] missing connection phenotype patients) outcome, poses extra challenge. classical potentially effect nutrients metabolites growth, completely cytotoxicity similar efficacy other CGTs. All things considered, that, eventually prevail CGT, ability incorporate simpler unconventional descriptions allows users desired prescriptive element 4.0.Naturally, every technique, acceptable validate depending model's importance found.[60] Active discussions filed indicate establish good practice (GMP) compliance.[61, 62] definition suitable regulation technical question addressed collaboration parties concerned.5ConclusionsWe conclude robust products: measurements execution well-characterised adaptive methods. cohesive multidisciplinary combining profound underlying biology engineering skills identify relationships compile twin. Considering problem hand, wish highlight management experts very diverse fields. Product design, bioprocesses, development, sciences, end regulators come together guide meaningful concepts applicable products, order therapeutic options vast number patients.

Language: Английский

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Nanotechnology revolutionizing osteosarcoma treatment: Advances in targeted kinase inhibitors

Nanotechnology Reviews, Journal Year: 2025, Volume and Issue: 14(1)

Published: Jan. 1, 2025

Abstract Osteosarcoma (OS) is the most frequent primary malignant bone tumor in adolescents and young adults. Despite advances therapy, OS remains an ominous problem because of its high metastatic potential, resistance to standard great physical, psychological, financial burden on patients. Available treatment options like surgery high-dose chemotherapy are limited by chemotoxicity, multimed resistance, adverse effects quality life Extrapolated from wide array vitro vivo studies, application kinase inhibitors targeting oncogenic signaling pathways, such as insulin-like growth factor 1 receptor, PDGFR, PI3K/AKT/mTOR pathway, appears quite promising. However, patients plagued with challenges poor bioavailability, off-target effects, mechanisms, which prevent clinical application. This review explores how nanotechnology beginning meet these challenges. Liposomes, polymeric nanoparticles, metallic nanoparticles among that provide new solutions for delivery bioavailability inhibitors, reducing systemic toxicity enhancing therapeutic accuracy. Active or passive enabled nanocarriers, enable drugs specifically act tissues while minimizing healthy cells. Additionally, diagnostic functionalities combined into theranostic platforms through pave way personalized medicine approaches. Nanoparticle-based have shown efficacy preclinical setting overcome drug improve targeting, sustained release drug. These dramatic improving outcomes at much less than currently available treatments. shows need further exploration bridge exciting findings practice. Future studies should seek optimize nanoparticle design evade enhance target specificity, reduce time-dependent toxicity. Further, incorporation a strategy has possibility changing treated bringing promise better patient life.

Language: Английский

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Next frontier in cancer treatment: an iPSC supported CAR-T cell therapies

Asia-Pacific Journal of Oncology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 27, 2024

The spread of cancer and huge number related deaths has made it imperative for scientist to search new diagnostic, prognostic treatment tools its effective management. Chimeric Antigen Receptor (CAR) T-cell (CAR-T) therapy been brought into limelight as novel strategy cancer. In this review we briefly introduced the CAR-T highlighting successful application various cancers along with possibility integrating induced pluripotent stem cell (iPSC) technology overcome intrinsic limitation. mechanism is based on isolation white blood cells through leukapheresis from patients’ or healthy donors followed by T-cells (a type immune cells). isolated are engineered using a viral vector other methods incorporate CAR gene expanded in lab. This modification enables T distinguish attack antigens surfaces cells. However, approach not without inherent limitations. notable limitation exhaustible availability within donor patient system, which can restrict feasibility effectiveness therapeutic intervention. A potential solution may lie synergizing technology. iPSC lies ability provide an endless supply resources. substantial amount research revealed production human Therefore, such combinatorial utilizing mounting immunological against continues iPSCs prove revolutionary designing pragmatic treatment.

Language: Английский

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