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IPSC nanovesicles cardiac cell repair

ABSTRACT: Here, we engineered functional EV-like nanovesicles (NVs) from human-induced pluripotent stem cells and highlight their potential as a functional surrogate for natural EVs for tissue repair. Proteome profiling of target cells further identified pathways implicated in angiogenesis (MYH10, AAMP, SLC3A2), cell survival (CCN2, RHEB, IGF2R, MFGE8), and fibroblast activation (CCN2, STAT3, COL1A1/2/4A2/12A1, ACTN1/4, ACTC1). This study demonstrates scalable generation of functional NVs and highlights their potential as a functional surrogate for natural EVs for tissue repair.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Heart, Multipotent Stem Cell

DISEASE(S): Cardiovascular System Disease

SUBMITTER: david greening

LAB HEAD: David Greening

PROVIDER: PXD036654 | Pride | 2023-01-15

REPOSITORIES: pride

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			Action	DRS
	Angio_CE_1.raw	Raw
	Angio_CE_2.raw	Raw
	Angio_CE_3.raw	Raw
	Angio_CE_4.raw	Raw
	Angio_CL_1.raw	Raw

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Publications

Scalable Generation of Nanovesicles from Human-Induced Pluripotent Stem Cells for Cardiac Repair.

Lozano Jonathan J Rai Alin A Lees Jarmon G JG Fang Haoyun H Claridge Bethany B Lim Shiang Y SY Greening David W DW

International journal of molecular sciences 20221118 22

Extracellular vesicles (EVs) from stem cells have shown significant therapeutic potential to repair injured cardiac tissues and regulate pathological fibrosis. However, scalable generation of stem cells and derived EVs for clinical utility remains a huge technical challenge. Here, we report a rapid size-based extrusion strategy to generate EV-like membranous nanovesicles (NVs) from easily sourced human iPSCs in large quantities (yield 900× natural EVs). NVs isolated using density-gradient separa ...[more]

PMID: 36430812

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Functional nanovesicles from human trophectodermal cells for embryo implantation

Project description:Extracellular vesicles (EVs) are important mediators of embryo attachment and outgrowth critical for successful implantation. While EVs have garnered immense interest in their therapeutic potential in assisted reproductive technology by improving implantation success, their large-scale generation remains a major challenge. Here, we report a rapid and scalable production of nanovesicles (NVs) from human trophectoderm cells (hTSCs) via membrane extrusion; these NVs can be generated in approximately 6 hours with a 20-fold higher yield than EVs isolated from the same number of cells. NVs display similar biophysical traits (morphologically intact, spherical, 90-130 nm) to EVs, and are laden with hallmark players of implantation (ITGA2/V, ITGB1, PRDX1, SOD2, and MFGE8) that include cell-matrix adhesion and extracellular matrix organisation proteins. Functionally, NVs are readily taken up by low-receptive endometrial HEC1A cells and reprogram their proteome towards a receptive phenotype that support hTSC spheroid attachment. Moreover, a single dose treatment of NVs significantly enhanced adhesion and spreading of mouse embryo trophoblast on fibronectin matrix. We show functional potential of NVs in enhancing embryo implantation and highlight their rapid and scalable generation, amenable to clinical utility.

2023-02-20 | PXD038856 | Pride

Transcriptional profiling of human fibroblasts cells from normal, PD patient, and PD patient treatment with mouse cavernous pericyte-derived extracellular vesicles-mimetic nanovesicles (PC-NVs)

Project description:Peyronie’s disease (PD) is a severe fibrotic diseases in the tunica albuginea that causes penis curvatures and leads to penile pain, deformity as well as erectile dysfunction (ED). The role of pericytes in the pathogenesis of fibrosis have been perceived recently. Extracellular vesicles (EVs)-mimetic nanovesicles (NVs) have emerged particular interest in intercellular communication between cells in various fields of fibrosis disease. However, the global gene expression of pericytes-derived EVs-mimetic NVs in regulating fibrosis remains unknown. Here we used RNA-sequencing technology to investigate the potential target genes regulated by pericytes-derived EVs-mimetic NVs in primary fibroblasts derived from human PD plaque.

2021-07-07 | GSE146500 | GEO

culture media formulations on extracellular vesicle production by amniotic epithelial cells

Project description:Abstract The therapeutic properties of extracellular vesicles (EVs) derived from stem cells and stem-like cells make them promising cell-free alternative to regenerative medicine. However, clinical translation of this technology relies heavily on the ability to manufacture EVs in a scalable, reproducible, cGMP-compliant manner. The choice of cell culture media is a critical component for EV manufacturing. In this study, we used human amniotic epithelial cells (hAECs) as a cell model system to explore the effect of chemically defined serum-free media on EV production. Here we showed that different culture media and different cell culture supplements have variable effects on EV production including culture parameters, EV yield, and EV biogenesis. Cell viability and proliferation rate are not reliable quality indicators of EV manufacturing. The levels of common EV tetraspanins and epitope makers can be impacted by different culture media formulations even when culture parameters are maintained, and EV yield are comparable. This study has uncovered some critical aspects regarding EV production culture media that need to be considered in clinical-grade scalable EV manufacturing.

2021-12-20 | PXD024733 | Pride

RBM7 regulates breast cancer progression

Project description:In order to profile the alternative splicing events that were regulated by RBM7, RNA-seq was performed on MDA-MB-231 cells which was used to generate RBM7 stably depleted breast cancer cells. And to explore how RBM7 regulated splicing target MFGE8, MDA-MB-231 cells stably overexpressed respectively MFGE8 long isoform (MFGE8-L) and short isoform (MFGE8-S) that were used for RNA-sequence.

2024-07-10 | GSE248938 | GEO

Pericyte-derived extracellular vesicle-mimetic nanovesicles ameliorates erectile dysfunction through Lipocalin 2 in the diabetic mice

Project description:Diabetes mellitus is one of the main causes of erectile dysfunction (ED). Due to neurovascular dysfunction, men with diabetic ED do not respond well to oral phosphodiesterase 5 inhibitors. Pericyte-derived extracellular vesicles-mimetic nanovesicles (PC-NVs) are known to play a role in promoting nerve regeneration in a mouse model of cavernous nerve injury. Here, we report that administration of PC-NVs can effectively promote penile angiogenesis and neural regeneration, thereby improving erectile function under diabetic conditions. We found that PC-NVs can induce endothelial proliferation, migration, and reduce cell apoptosis under diabetic conditions. In addition, PC-NVs induce neural regeneration in STZ-induced diabetic mice in vivo and ex vivo MPG or DRG explants under high-glucose conditions. We found that Lipocalin 2 (Lcn2) is a new target of PC-NVs in this process, indicating that PC-NVs exert their angiogenesis and nerve regeneration effects by activating MAP kinase, PI3K/Akt and suppressing P53 signaling pathway in an Lcn2-dependent manner under diabetic conditions. Our findings provide new conclusive evidence that PC-NVs mediate neurovascular regeneration and erectile function recovery under diabetic conditions through an Lcn2-dependent mechanism. Local administration of PC-NVs may be a promising treatment strategy for the treatment of diabetic ED.

2022-07-13 | GSE183220 | GEO

Independent human mesenchymal stromal cell-derived extracellular vesicle preparations differentially attenuate symptoms in an advanced murine Graft-versus-Host-Disease model

Project description:Extracellular vesicles (EVs) harvested from conditioned media of human mesenchymal stromal cells (MSCs) suppress acute inflammation in various disease models and promote regeneration of damaged tissues. Following successful treatment of an acute steroid-refractory Graft-versus-Host disease (GvHD) patient with EVs prepared from conditioned media of human bone marrow-derived MSCs, we focus on improving the MSC-EV production for the clinical application. Independent MSC-EV preparations all produced according to a standardized procedure, reveal broad immunomodulatory differences. Only a proportion of our MSC-EV products effectively modulate immune responses in a multi-donor mixed lymphocyte reaction (mdMLR) assay. To explore the relevance of such differences, we have established an optimized mouse GvHD model. The functional testing of selected MSC-EV preparations demonstrate that MSC-EV preparations revealing immunomodulatory capabilities in the mdMLR assay also effectively suppress GvHD symptoms in this model. In contrast, MSC-EV preparations, lacking such in vitro activities, also fail to modulate GvHD symptoms in vivo. Searching for differences of the active and inactive MSC-EV preparations, we failed to identify concrete proteins or miRNAs that could serve as surrogate markers. Thus, standardized MSC-EV production strategies may not be sufficient to warrant manufacturing of MSC-EV products with reproducible qualities. Consequently, given this functional heterogeneity, every individual MSC-EV preparation considered for the clinical application should be evaluated for its therapeutic potency prior to administration to patients. Here, we qualified the mdMLR assay for such analyses.

2023-06-12 | PXD032744 | Pride

Gene regulation in gastrocnemius muscle of denervated mouse with HDAC4 inducible knockout or class IIa HDAC inhibitior NVS-HD1 treatment

Project description:Our class IIa HDAC inhibitor, NVS-HD1, inhibited HDAC4 with less than 1 nM potency while exhibiting >200 fold selectivity on class IIa HDACs compared to class I (HDAC1, 3, 8) and class IIb (HDAC6) HDACs, making it the most potent and selective class IIa HDAC inhibitor reported so far. We tested the efficacy of NVS-HD1 in the mouse denervation model, either alone or on the genetic background of HDAC4 whole-body inducible knockout (HDAC4 iRKO). Global gene expression changes in gastrocnemius muscles were profiled by RNAseq. In the innervated control legs, HDAC4 knockout or NVS-HD1 caused little changes in gene expression compared to WT mice. HDAC4 knockout or NVS-HD1 mainly reversed denervation induced changes and the genes regulated by them largely overlap, suggesting that NVS-HD1 is quite specific against class IIa HDACs.

2019-11-27 | GSE136627 | GEO

RNA-Seq profiling of ERCC1-XPF DNA repair defect (Er1F/-) versus wild type (Er1F/+) macrophages

Project description:DNA damage and metabolic disorders are intimately linked with premature disease onset but the underlying mechanisms remain poorly understood. Persistent DNA damage accumulation in tissue-infiltrating macrophages carrying an ERCC1-XPF DNA repair defect (Er1F/-) riggers Golgi dispersal, dilation of endoplasmic reticulum, autophagy and exosome biogenesis leading to the secretion of extracellular vesicles (EVs) in vivo and ex vivo.

2019-11-04 | E-MTAB-8439 | biostudies-arrayexpress

hUC-MSCs-derived MFGE8 ameliorates locomotor dysfunction via inhibition of ITGB3/NF-κB signaling in an NMO mouse model

Project description:Neuromyelitis optica (NMO) is a severe autoimmune inflammatory disease of the central nervous system that affects motor function and causes relapsing disability. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) have been used extensively in the treatment of various inflammatory diseases, due to their potent regulatory roles that can mitigate inflammation and repair damaged tissues. However, their use in NMO is currently limited, and the mechanism underlying the beneficial effects of hUC-MSCs on motor function in NMO remains unclear. In this study, we investigate the effects of hUC-MSCs on the recovery of motor function in an NMO systemic model. Our findings demonstrate that milk fat globule epidermal growth 8 (MFGE8), a key functional factor secreted by hUC-MSCs, plays a critical role in ameliorating motor impairments. We also elucidate that the MFGE8/Integrin αvβ3/NF-κB signaling pathway is partially responsible for structural and functional recovery, in addition to motor functional enhancements induced by hUC-MSC exposure. Taken together, these findings strongly support the involvement of MFGE8 in mediating hUC-MSCs-induced improvements in motor functional recovery in an NMO mouse model. In addition, this provides new insight on the therapeutic potential of hUC-MSCs and the mechanisms underlying their beneficial effects in NMO.

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