Development of a Robust Consensus Modeling Approach for Identifying Cellular and Media Metabolites Predictive of Mesenchymal Stromal Cell Potency DOI Creative Commons
Alexandria Van Grouw, Maxwell B. Colonna, Ty S. Maughon

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Feb. 4, 2023

Abstract Mesenchymal stromal cells (MSCs) have shown promise in regenerative medicine applications due part to their ability modulate immune cells. However, MSCs demonstrate significant functional heterogeneity terms of immunomodulatory function because differences MSC donor/tissue source, as well non-standardized manufacturing approaches. As metabolism plays a critical role expand therapeutic numbers ex vivo , we comprehensively profiled intracellular and extracellular metabolites throughout the expansion process identify predictors (T cell modulation indoleamine-2,3-dehydrogenase (IDO) activity). Here, media non-destructive manner through daily sampling nuclear magnetic resonance (NMR), at end using mass spectrometry (MS). Using robust consensus machine learning approach, were able panels predictive for 10 independent lines. This approach consisted identifying 2 or more models then building based on these metabolite panels. Consensus with high value included multiple lipid classes (such phosphatidylcholines, phosphatidylethanolamines, sphingomyelins) while proline, phenylalanine, pyruvate. Pathway enrichment identified metabolic pathways significantly associated such sphingolipid signaling metabolism, arginine proline autophagy. Overall, this work establishes generalizable framework that predict function, guiding future efforts identification potency lines engineering.

Language: Английский

Biomarkers for osteoarthritis: Current status and future prospects DOI Creative Commons
Ali Mobasheri, Christian S. Thudium, Anne‐Christine Bay‐Jensen

et al.

Best Practice & Research Clinical Rheumatology, Journal Year: 2023, Volume and Issue: 37(2), P. 101852 - 101852

Published: June 1, 2023

Osteoarthritis (OA) is the most common form of arthritis globally and a major cause pain, physical disability, loss economic productivity, with currently no causal treatment available. This review article focuses on current research OA biomarkers potential for using in future clinical practice trials investigational drugs. We discuss how biomarkers, specifically soluble ones, have long path to go before reaching standards care. also can help phenotyping subtyping achieve enhanced stratification move toward better-designed trials. describe be used molecular endotyping determining outcomes cell-based therapies. Biomarkers developed as surrogate end points private-public consortia biotechnology pharmaceutical industries develop more effective targeted personalized treatments enhance care patients OA.

Language: Английский

Citations

37
The enhancer landscape predetermines the skeletal regeneration capacity of stromal cells DOI Open Access
Sarah Hochmann, Kristy Ou, Rodolphe Poupardin

et al.

Science Translational Medicine, Journal Year: 2023, Volume and Issue: 15(688)

Published: March 22, 2023

Multipotent stromal cells are considered attractive sources for cell therapy and tissue engineering. Despite numerous experimental clinical studies, broad application of therapeutics is not yet emerging. A major challenge the functional diversity available sources. Here, we investigated regenerative potential clinically relevant human from bone marrow (BMSCs), white adipose tissue, umbilical cord compared with mature chondrocytes skin fibroblasts in vitro vivo. Although all types could express transcription factors related to endochondral ossification, only BMSCs formed cartilage discs that fully regenerated critical-size femoral defects after transplantation into mice. We identified type–specific epigenetic landscapes as underlying molecular mechanism controlling transcriptional differentiation networks. Binding sites commonly expressed enhancer promoter regions ossification-related genes, including Runt bZIP families, were accessible but extraskeletal cells. This suggests an epigenetically predetermined depending on origin allows common trigger distinct organ-specific programs, facilitating forward selection regeneration-competent Last, demonstrate viable initiated defect healing through secretion osteopontin contributed transient mineralized hard callus formation immunodeficient mice, which was eventually replaced by murine recipient during final remodeling.

Language: Английский

Citations

26
Individual immune cell and cytokine profiles determine platelet-rich plasma composition DOI Creative Commons
Marcel Niemann,

Melanie Ort,

Luis Lauterbach

et al.

Arthritis Research & Therapy, Journal Year: 2023, Volume and Issue: 25(1)

Published: Jan. 10, 2023

Platelet-rich plasma (PRP) therapy is increasingly popular to treat musculoskeletal diseases, including tendinopathies and osteoarthritis (OA). To date, it remains unclear which extent PRP compositions are determined by the immune cell cytokine profile of individuals or preparation method. investigate this, we compared leukocyte distributions different products donor blood samples assessed effect pro-inflammatory cytokines on chondrocytes.

Language: Английский

Citations

15
Lyophilized human platelet lysate: manufacturing, quality control, and application DOI Creative Commons

Kerstin Wendland,

Lea Koblin,

Dirk Stobbe

et al.

Frontiers in Cell and Developmental Biology, Journal Year: 2025, Volume and Issue: 13

Published: Jan. 27, 2025

Background A significant number of platelet concentrates (PCs) is discarded daily in blood banks due to limited shelf life. Human lysate (HPL), derived from expired PCs, has gained attention as an ethical and sustainable cell culture media supplement biomedical research therapy production. However, HPL subject decisive disadvantages such batch differences lack storage stability. To overcome these limitations enhance the applicability HPL, we developed manufacturing protocol including a lyophilization process. The aim this study was investigate influence on parameters quality control, growth factor concentrations human mesenchymal stromal cells (hMSCs). Methods We performed paired comparison six batches lyophilized (L-HPL) regarding pH, total protein, osmolality, sodium, potassium chloride concentration. Concentrations 11 factors cytokines were compared between L-HPL. Additionally, determined yield, proliferation capacity, viability trilineage differentiation potential hMSCs following expansion HPL- L-HPL-supplemented media. Results Quantification revealed non-altered osmolality slightly lower sodium L-HPL HPL. Growth cytokine did not differ Cell division cycles cultured either or L-HPL-containing comparable. Cells differentiated medium containing showed higher capacity for osteogenic differentiation, while adipogenic chondrogenic potentials remained unchanged. Conclusion successfully method produce well-applicable with reveal any relevant control routine testing, hMSC functionality, demonstrating suitability supplement. These results emphasize alternative animal-derived serum products drug development.

Language: Английский

Citations

0
Toward Machine Learning Electrospray Ionization Sensitivity Prediction for Semiquantitative Lipidomics in Stem Cells DOI Creative Commons
Alexandria Van Grouw, Markace A. Rainey,

Olivia K. Reid

et al.

Journal of Chemical Information and Modeling, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 5, 2025

Specificity, sensitivity, and high metabolite coverage make mass spectrometry (MS) one of the most valuable tools in metabolomics lipidomics. However, translation MS methods to multiyear studies conducted across multiple batches is limited by variability electrospray ionization response, making batch-to-batch comparisons challenging. This limitation creates an artificial divide between nontargeted discovery work that broad scope but terms absolute quantitation ability targeted highly accurate due need for matched isotopically labeled standards. These issues are often observed stem cell using metabolomic lipidomic approaches, where patient recruitment can be a years-long process samples become available discrete every few months. To bridge this gap, we developed machine learning model predicts sensitivity lipid classes have shown correlation with potency. Molecular descriptors derived from these lipids' chemical structures used as input predict enabling moderate accuracy (semiquantitation). Model performance was evaluated via internal external validation cultured cells various donors, achieving global percent errors 40% 20% positive negative ion modes, respectively. Although typically insufficient traditional lipidomics experiments, it sufficient semiquantitative estimation marker concentrations without specific standards many times unavailable. Furthermore, precision model-predicted 16.9% mode 7.5% mode, indicating promise data harmonization batches. The set molecular models described here able yield higher than those previously published literature, showing toward

Language: Английский

Citations

0
Humanising nanotoxicology: replacement of animal-derived products in the application of integrated approaches to testing and assessment of nanomaterial inhalation hazard DOI Creative Commons

Russell Fraser,

KC Campbell,

Pawel Pokorski

et al.

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

Published: Feb. 12, 2025

Over the past decade, development of nanomaterials (NMs) has surged, highlighting their potential benefits across multiple industries. However, concerns regarding human and environmental exposure remain significant. Traditional in vivo models for safety assessments are increasingly viewed as unfeasible unethical due to diverse forms biological effects NMs. This prompted design Novel Approach Methods (NAMs) streamline risk assessment predict hazards without relying on animal testing. A critical aspect advancing NAMs is urgent need replace animal-derived products assay protocols. Incorporating or synthetic alternatives can significantly reduce ethical burden use while enhancing relevance toxicity study evaluates impact removing from standard acellular vitro assays recommended a published Integrated Approaches Testing Assessment (IATA) inhaled We specifically assessed replacing fetal bovine serum with platelet lysate reactivity tests testing using panel well-characterized Significant differences NM dramatic changes A549 cell growth rates responses NMs were observed under different media conditions. Our findings demonstrate that variations experimental setup fundamentally hazard assessment, influencing interpretation results within specific tiered strategies. Further investigation needed support shift toward more does not rely materials.

Language: Английский

Citations

0
Enhance the therapeutic efficacy of human umbilical cord-derived mesenchymal stem cells in prevention of acute graft-versus-host disease through CRISPLD2 modulation DOI Creative Commons
Qing Xu, Rui Wang, Ke Sui

et al.

Stem Cell Research & Therapy, Journal Year: 2025, Volume and Issue: 16(1)

Published: May 1, 2025

Language: Английский

Citations

0
Genetic engineering of primary human mesenchymal stromal cells using the CRISPR/Cas9 synergistic activation mediator DOI

Madeleine C. Killer,

Xiaoqing Zhang, Till Adhikary

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: May 12, 2025

Abstract Background: Genomic editing technologies provide rich opportunities to approach inherited human diseases, but gene delivery strategies are still challenging. While engineered induced pluripotent stem cells or adeno-associated viral vectors may be associated with severe adverse effects, mesenchymal stromal (MSC) have abundantly been used for various cell therapeutic and proven safe in a large number of clinical trials. MSC can expanded ex vivoon scale, genetic engineering these somatic therefore offer an alternative mode delivery. Methods: We applied the guide RNA-directed CRISPR/Cas9 synergistic activation mediator (SAM) technology immortalized (iMSC) primary (pMSC) order activate expression CEBPA CIITA. Results: After meticulous protocol optimization, both genes were successfully MSC. For CIITA induction, we confirmed downstream target HLA-DR on mRNA protein level. Unlike adipogenic, chondrogenic osteogenic genes, which naturally upon in vitro differentiation, HLA class II molecules not constitutively expressed Lentiviral transposon-based applied, only lentiviral transduction SAM sequences, CEBPA, CIITA, HLA-DR HLA-DR achieved pMSC. Conclusion: guided system was capable induce transcriptionally silent iMSC The resulting genetically suitable promising cell-based strategies.

Language: Английский

Citations

0
The benefits of adipocyte metabolism in bone health and regeneration DOI Creative Commons
Lisa-Marie Burkhardt, Christian H. Bucher,

Julia Löffler

et al.

Frontiers in Cell and Developmental Biology, Journal Year: 2023, Volume and Issue: 11

Published: Feb. 21, 2023

Patients suffering from musculoskeletal diseases must cope with a diminished quality of life and an increased burden on medical expenses. The interaction immune cells mesenchymal stromal during bone regeneration is one the key requirements for restoration skeletal integrity. While osteo-chondral lineage support regeneration, excessive accumulation adipogenic thought to promote low-grade inflammation impair regeneration. Increasing evidence indicates that pro-inflammatory signaling adipocytes responsible various chronic diseases. This review aims summarize features marrow by phenotype, function, secretory features, metabolic properties their impact formation. In detail, master regulator adipogenesis prominent diabetes drug target, peroxisome proliferator-activated receptor γ (PPARG), will be debated as potential therapeutic approach enhance We explore possibilities using clinically established PPARG agonists, thiazolidinediones (TZDs), treatment strategy guide induction pro-regenerative, metabolically active adipose tissue. this induced tissue type providing necessary metabolites sustain osteogenic-as well beneficial fracture healing highlighted.

Language: Английский

Citations

9
Enhancer hijacking at the ARHGAP36 locus is associated with connective tissue to bone transformation DOI Creative Commons
Uirá Souto Melo, Jerome Jatzlau, Cesar A. Prada‐Medina

et al.

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: April 11, 2023

Heterotopic ossification is a disorder caused by abnormal mineralization of soft tissues in which signaling pathways such as BMP, TGFβ and WNT are known key players driving ectopic bone formation. Identifying novel genes related to the process important steps for future gene therapy disorders. In this study, we detect an inter-chromosomal insertional duplication female proband disrupting topologically associating domain causing ultra-rare progressive form heterotopic ossification. This structural variant lead enhancer hijacking misexpression ARHGAP36 fibroblasts, validated here orthogonal vitro studies. addition, overexpression inhibits TGFβ, activates hedgehog genes/proteins extracellular matrix production. Our work on genetic cause case has revealed that plays role formation metabolism, outlining first details contributing bone-formation -disease.

Language: Английский

Citations

8