Evidence map›Paper›PMID 39949490›Full record

ArticleCellular and molecular bioengineering2025

CRISPR-dCas9 Activation of TSG-6 in MSCs Modulates the Cargo of MSC-Derived Extracellular Vesicles and Attenuates Inflammatory Responses in Human Intervertebral Disc Cells In Vitro.

Iker Martinez-Zalbidea, Gabbie Wagner, Nea Bergendahl, Addisu Mesfin, Varun Puvanesarajah, Wolfgang Hitzl, Stefan Schulze, Karin Wuertz-Kozak

Abstract read
In one paragraph

Article in Cellular and molecular bioengineering, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.

0numbers the graph read from it
0cells of the map it votes in
14citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

14 citing papers in PubMed.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Iker Martinez-ZalbideaDepartment of Biomedical Engineering, Rochester Institute of Technology (RIT), Rochester, NY USA.ORCID 0009-0005-2354-1058
Gabbie WagnerDepartment of Biomedical Engineering, Rochester Institute of Technology (RIT), Rochester, NY USA.ORCID 0009-0000-9195-4114
Nea BergendahlDepartment of Biomedical Engineering, Rochester Institute of Technology (RIT), Rochester, NY USA.
Addisu MesfinMedstar Orthopaedic Institute, Georgetown University School of Medicine Washington, Washington, DC USA.ORCID 0000-0003-0076-4185
Varun PuvanesarajahDepartment of Orthopedics and Rehabilitation, University of Rochester Medical Center, Rochester, NY USA.ORCID 0000-0002-3296-6403
Wolfgang HitzlResearch and Innovation Management (RIM), Paracelsus Medical University, Salzburg, Austria.ORCID 0000-0002-7696-1479
Stefan SchulzeThomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY USA.ORCID 0000-0002-4771-7987
Karin Wuertz-KozakDepartment of Biomedical Engineering, Rochester Institute of Technology (RIT), Rochester, NY USA.ORCID 0000-0002-3281-4629

Funding

Extracellular vesicles produced by CRISPR-activated MSCs: A potential therapy for degenerative disc diseaseR21AR081987 · NIAMS · ROCHESTER INSTITUTE OF TECHNOLOGY · PI WUERTZ-KOZAK, KARIN · 2023 to 2024
$365k
NIAMS NIH HHS R21 AR081987
6 · The paper itself

Abstract

Purpose: The purpose of this study was to boost the therapeutic effect of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) by overexpressing the gene TSG-6 through CRISPR activation, and assess the biological activity of EVs from these modified MSCs Methods: An immortalized human MSC line was transduced with a CRISPR activation lentivirus system targeting TSG-6. MSC-EVs were harvested by ultracentrifugation and particle number/size distribution was determined by nanoparticle tracking analysis. The efficiency of transduction activation was assessed by analyzing gene and protein expression. EV proteomic contents were analyzed by mass spectrometry. Human IVD cells from patients undergoing spinal surgery were isolated, expanded, exposed to IL-1β pre-stimulation and co-treated with MSC-EVs. Results: MSC-EVs presented size distribution, morphology, and molecular markers consistent with common EV characteristics. The expression level of TSG-6 was significantly higher (> 800 fold) in transduced MSCs relative to controls. Protein analysis of MSCs and EVs showed higher protein expression of TSG-6 in CRISPR activated samples than controls. Proteomics of EVs identified 35 proteins (including TSG-6) that were differentially expressed in TSG-6 activated EVs vs control EVs. EV co-Treatment of IL-1β pre-Stimulated IVD cells resulted in a significant downregulation of IL-8 and COX-2. Conclusions: We successfully generated an MSC line overexpressing TSG-6. Furthermore, we show that EVs isolated from these modified MSCs have the potential to attenuate the pro-inflammatory gene expression in IVD cells. This genomic engineering approach hence holds promise for boosting the therapeutic effects of EVs. Supplementary Information: The online version contains supplementary material available at 10.1007/s12195-025-00843-4.

Indexed as

ExosomesGenomic engineeringIVDProteomicsRegenerationTranscriptomic modulation

Identifiers

PMID39949490
PMCPMC11813855

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.