Machine Learning and Metabolomics Predict Mesenchymal Stem Cell Osteogenic Differentiation in 2D and 3D Cultures DOI Creative Commons
Michail E. Klontzas, Spyros I. Vernardis,

Aristea Batsali

et al.

Journal of Functional Biomaterials, Journal Year: 2024, Volume and Issue: 15(12), P. 367 - 367

Published: Dec. 5, 2024

Stem cells have been widely used to produce artificial bone grafts. Nonetheless, the variability in degree of stem cell differentiation is an inherent drawback graft development and requires robust evaluation tools that can certify quality cell-based products avoid source-tissue-related patient-specific outcomes. Omics analyses utilised for attributes all stages biomanufacturing. Herein, metabolomics combination with machine learning was benchmarking osteogenic 2D 3D cultures. Metabolomics analysis performed use gas chromatography–mass spectrometry (GC-MS). A set 11 metabolites train XGboost model which achieved excellent performance distinguishing between differentiated undifferentiated umbilical cord blood mesenchymal (UCB MSCs). The benchmarked against samples not present training set, being able efficiently capture osteogenesis UCB MSC cultures area under curve (AUC) 82.6%. On contrary, did any Wharton’s Jelly samples, are well-known underperformers (AUC 56.2%). Mineralisation significantly correlated levels fumarate, glycerol, myo-inositol, four found most important (R2 = 0.89, R2 0.94, 0.96, p 0.016, 0.0059, 0.0022, respectively). In conclusion, our results indicate be reliable potency assays Advanced Therapy Medicinal Products.

Language: Английский

Beneath the radar: immune-evasive cell sources for stroke therapy DOI Creative Commons
Beatriz Achón Buil, Nora H. Rentsch, R. Weber

et al.

Trends in Molecular Medicine, Journal Year: 2024, Volume and Issue: 30(3), P. 223 - 238

Published: Jan. 25, 2024

Stem cell therapy is an emerging treatment paradigm for stroke patients with remaining neurological deficits. While allogeneic transplants overcome the manufacturing constraints of autologous grafts, they can be rejected by recipient's immune system, which identifies foreign cells through human leukocyte antigen (HLA) system. The heterogeneity HLA molecules in population would require a very high number lines, may still inadequate rare genetic HLAs. Here, we outline key progress engineering pluripotent stem and derived to evade host's reducing lines required, examine safety measures explored both preclinical studies upcoming clinical trials.

Language: Английский

Citations

10
Biomarkers of Efficacy and Safety of the Academic BCMA-CART ARI0002h for the Treatment of Refractory Multiple Myeloma DOI
Aina Oliver‐Caldés, Marta Español‐Rego, Aintzane Zabaleta

et al.

Clinical Cancer Research, Journal Year: 2024, Volume and Issue: 30(10), P. 2085 - 2096

Published: March 11, 2024

Abstract Purpose: B-cell maturation antigen (BCMA)-chimeric receptor T-cells (CART) improve results obtained with conventional therapy in the treatment of relapsed/refractory multiple myeloma. However, high demand and expensive costs associated CART might prove unsustainable for health systems. Academic CARTs could potentially overcome these issues. Moreover, response biomarkers resistance mechanisms need to be identified addressed efficacy patient selection. Here, we present clinical ancillary 60 patients treated academic BCMA-CART, ARI0002h, CARTBCMA-HCB-01 trial. Patients Methods: We collected apheresis, final product, peripheral blood bone marrow samples before after infusion. assessed BCMA, T-cell subsets, kinetics antibodies, aplasia, cytokines, measurable residual disease by next-generation flow cytometry, correlated outcomes. Results: At cut-off date March 17, 2023, a median follow-up 23.1 months (95% CI, 9.2–37.1), overall rate first 3 was 95% [95% confidence interval (CI), 89.5–100]; cytokine release syndrome (CRS) observed 90% (5% grades ≥3) grade 1 immune effector cell-associated neurotoxicity reported 2 (3%). Median progression-free survival 15.8 11.5–22.4). Surface BCMA not predictive or survival, but soluble worse outcomes CRS severity. Activation marker HLA-DR apheresis longer increased exhaustion markers poorer ARI0002h loss aplasia were relapse. Conclusions: Despite deep sustained responses achieved several that correlate poor

Language: Английский

Citations

7
Mesenchymal Stromal Cell-Based Products: Challenges and Clinical Therapeutic Options DOI Open Access
Débora B. Mello, Fernanda Mesquita, Danúbia Silva dos Santos

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(11), P. 6063 - 6063

Published: May 31, 2024

Mesenchymal stromal cell (MSC)-based advanced therapy medicinal products (ATMPs) are being tried in a vast range of clinical applications. These cells can be isolated from different donor tissues by using several methods, or they even derived induced pluripotent stem embryonic cells. However, ATMP heterogeneity may impact product identity and potency, and, consequently, trial outcomes. In this review, we discuss these topics the need to establish minimal criteria regarding manufacturing MSCs so that innovative therapeutics better positioned contribute advancement regenerative medicine.

Language: Английский

Citations

6
A Quantitative Approach to Potency Testing for Chimeric Antigen Receptor-Encoding Lentiviral Vectors and Autologous CAR-T Cell Products, Using Flow Cytometry DOI Creative Commons
Juan José Mata, Letícia Alserawan,

Carolina España

et al.

Pharmaceutics, Journal Year: 2025, Volume and Issue: 17(3), P. 303 - 303

Published: Feb. 25, 2025

Background/Objectives: Potency testing of clinical-grade lentiviral vectors (LVVs) is critical to support a drug’s commercial approval. Careful consideration should be paid the development suitable potency test during clinical development. We aimed develop an affordable, quantitative for our CAR19-LVV, based on measure transgene’s functional activity. Methods: Several indicators activity CAR19-LVV were explored in co-culture setting CAR-transduced Jurkat cells and CD19-expressing target cells. The selected assay was further developed subjected validation. Assay’s adaptability other CAR-encoding LVV autologous CAR-T cell products also investigated. Results: Measure CD69 expression membrane Jurkat-CAR-expressing specific indicator CAR functionality. Quantification terms mean fluorescence intensity (MFI), coupled with intra-assay standard curve calibration, allows high precision, specificity, robustness, linearity accuracy. has shown optimal performance CARBCMA-LVV product. Importantly, we show that primary T cells, reflects cytotoxicity. After adaptation, have applied CD69-based test, simultaneous measurement cytotoxicity, products, demonstrating assay’s specificity this context. Conclusions: validated, vitro cell-based using flow-cytometry method, CAR19-LVV. detection T-cell activation upon binding antigen, which transgene easily adapted another LVV, targeting different molecule. Furthermore, same principle can context products. shows reduced variability among compared IFNγ evaluation traditional semi-quantitative thereby directly evaluating mechanism action (MoA) assay.

Language: Английский

Citations

0
Callus organoids reveal distinct cartilage to bone transition mechanisms across donors and a role for biological sex DOI Creative Commons
Isaak Decoene, Hanna Svitina, Mohamed Belal Hamed

et al.

Bone Research, Journal Year: 2025, Volume and Issue: 13(1)

Published: March 26, 2025

Language: Английский

Citations

0
Advancing Potency Assay Development for Advanced Therapy Medicinal Products: A Comprehensive Approach and Regulatory Insights DOI
Alaa Abdellatif,

Mélissa Bou Jaoudeh,

Alex Zwiers

et al.

Human Gene Therapy, Journal Year: 2025, Volume and Issue: unknown

Published: April 21, 2025

The development of potency assays for Advanced Therapy Medicinal Products (ATMPs) presents significant challenges due to the variability starting materials and complex mechanisms action involved. This article aims address following key question: How can we design robust reliable ATMPs that accommodate product-specific align with evolving regulatory standards? To answer this, employed a mixed-methods approach, synthesizing data from scientific literature, industry reports, guidelines identify current limitations innovative solutions assay development. Our methodology integrates systematic review academic publications (2018-2024) capture recent advancements in biotechnology their applicability testing. We complemented this an analysis perspectives, drawn webinars white papers, as well detailed comparison global frameworks, including FDA's new guidance on assurance Cellular Gene (CGTs/ATMPs). Additionally, developed comprehensive database analyze used approved, rejected, withdrawn CGT/ATMP products, focusing technical challenges. Based multilevel analysis, propose framework designing, developing, validating different ATMP categories, taking into account unique constraints. also highlight emerging technologies, such droplet digital polymerase chain reaction reporter gene assays, tools improving precision reliability findings underscore need flexible, risk-based strategies evolve throughout product clinical trial phases. Future recommendations emphasize standardization, definition acceptable variability, stronger correlations between vitro outcomes.

Language: Английский

Citations

0
A novel multicolor fluorescent spot assay for the functional assessment of chimeric antigen receptor (CAR) T-cell products DOI
Djordje Atanackovic,

Thierry Iraguha,

Destiny Omili

et al.

Cytotherapy, Journal Year: 2024, Volume and Issue: 26(4), P. 318 - 324

Published: Feb. 10, 2024

Language: Английский

Citations

2
Potency assay to predict the anti-inflammatory capacity of a cell therapy product for macrophage-driven diseases: overcoming the challenges of assay development and validation DOI Creative Commons
Samar Sadeghi,

Laura Nimtz,

Elke Niebergall‐Roth

et al.

Cytotherapy, Journal Year: 2024, Volume and Issue: 26(5), P. 512 - 523

Published: Feb. 18, 2024

BackgroundGiven the high level of product complexity and limited regulatory guidance, designing implementing appropriate potency assays is often most challenging part establishing a quality control testing matrix for cell-based medicinal product. Among elusive tasks are selection suitable read-out parameters, development assay designs that closely model pathophysiological conditions, validation methods. Here we describe these challenges how they were addressed in developing an measures anti-inflammatory mesenchymal stromal cells (MSCs) M1 macrophage-dominated inflammatory environment.MethodsAn vitro inflammation was established by coculturing skin-derived ABCB5+ MSCs with THP-1 monocyte-derived M1-polarized macrophages. Readout amount interleukin 1 receptor antagonist (IL-1RA) secreted coculture, measured enzyme-linked immunosorbent assay.ResultsIL-1RA quantified guideline-concordant selectivity, accuracy precision over relevant concentration range. Consistent induction macrophage markers CD36 CD80 indicated successful differentiation polarization cells, which functionally confirmed release proinflammatory tumor necrosis factor α. Testing wide range MSC/macrophage ratios revealed optimal ratio near-maximal stimulation to secrete IL-1RA, providing absolute maximum levels per individual MSC can be used future comparison clinical efficacy. Batch 71 consecutively manufactured batches showed low overall failure rate comparability between donors.ConclusionsWe systematic therapeutically relevant, straightforward, robust reproducible measure immunomodulatory capacity macrophage-driven inflammation. The insights into may also helpful developers related other cellular functions indications.

Language: Английский

Citations

2
Production of donor-derived cytotoxic T lymphocytes with potent anti-leukemia activity for adoptive immunotherapy in high-risk pediatric patients given haploidentical hematopoietic stem cell transplantation DOI
Matteo Tanzi,

Enrica Montini,

Agnese Rumolo

et al.

Cytotherapy, Journal Year: 2024, Volume and Issue: 26(8), P. 878 - 889

Published: April 21, 2024

Language: Английский

Citations

1
Advancing CAR T-cell therapies: Preclinical insights and clinical translation for hematological malignancies DOI Creative Commons
Arun Kumar Arunachalam, Céline Gregoire, Beatriz Coutinho de Oliveira

et al.

Blood Reviews, Journal Year: 2024, Volume and Issue: unknown, P. 101241 - 101241

Published: Sept. 1, 2024

Language: Английский

Citations

1