LC Sciences – Technologies for Genomics & Proteomics Discoveries

Lactic acid bacteria naturally associated with ready-to-eat rocket salad can survive the human gastrointestinal transit

Admin — Tue, 07 May 2024 16:09:10 +0000

It was theorized that modernization and the decline in harmless microbial populations associated with food have altered the gut microbiota, impacting host metabolism and immunity. Western dietary patterns, characterized by processed foods and preservation methods, may significantly reduce the microbial population associated with food. To mitigate the consequences of bacterial deprivation, the integration of these diets with fermented foods is commonly proposed. Nonetheless, non-fermented food consumed raw may also be an important source of viable microbial cells for the human microbiome. In this study conducted by Università degli Studi di Milano, it investigates whether salad-associated LAB can survive the gastrointestinal transit (GIT) and contribute to the gut microbiota. They used LC Sciences’ 16S rRNA sequencing service on feces samples from the volunteers in the study. LAB strains were quantified and isolated from rocket salad (Eruca vesicaria subsp. sativa), and their survival through GIT was assessed via intervention trials in healthy adults and in vitro. Moreover, bacterial communities in fecal samples were analyzed after three days of rocket salad consumption. Washing with a sodium hypochlorite solution drastically reduced total bacterial load and eliminated viable LAB. The quantity of LAB introduced through salads did not significantly alter the gut microbiota composition. Rocket salads harbored Weissella and Leuconostoc species. A significant increase in Weissella spp. but not in Leuconostoc spp. was observed after the consumption of rocket salad. Simulated GIT experiments suggested that the food matrix and the initial number of ingested viable bacteria may have been important in determining survival. These findings propose that plant products could serve as sources of live LAB for the human gut. Further research with diverse vegetables and longer interventions is needed, encouraging studies on raw, non-fermented foods and their impact on the human intestinal microbiome.

Recovery on MRS agar (pH 6.7) of viable cells of the dominant lactic acid bacterial species from rocket salads in feces collected at the end of the three-day intervention trials

Panel A: Intervention with rocket salad R1 with (on the right, in green) or without (on the left, in red) washing using a sodium hypochlorite solution. Panel B: intervention with washed (on the right, in green) and unwashed (on the left, in red) rocket salad R2. Data are presented as the estimated minimum CFU count (eCFU; see the Methods section for details). Statistical analysis was performed using a Wilcoxon signed-rank test; *, p < 0.05; n.s., not significant.

Mantegazza Giacomo, Duncan Robin, Telesca Nicolò, Gargari Giorgio, Perotti Susanna, Riso Patrizia, Guglielmetti Simone. (2024) Lactic acid bacteria naturally associated with ready-to-eat rocket salad can survive the human gastrointestinal transit. Food Microbiology 118, 104418. [article]

Join LC Sciences at the AACR Annual Meeting – 2024

Admin — Wed, 13 Mar 2024 12:48:00 +0000

LC Sciences is thrilled to announce our participation in the upcoming American Association for Cancer Research
2024 annual meeting and conference
San Diego, CA- April 7th to April 10th!

Mark your calendars and come meet our team to discover how our innovative solutions are shaping the future of cancer research. Let’s collaborate, inspire, and drive progress towards a world free from cancer.

See you in San Diego!

We will be located in Booth 647

K235 acetylation couples with PSPC1 to regulate the m6A demethylation activity of ALKBH5 and tumorigenesis

Admin — Mon, 04 Mar 2024 20:02:52 +0000

N6-methyladenosine (m⁶A) modification plays important roles in bioprocesses and diseases. AlkB homolog 5 (ALKBH5) is one of two m⁶A demethylases. In this study led by Third Affiliated Hospital of Guangzhou Medical University, they reveal that ALKBH5 is acetylated at lysine 235 (K235) by lysine acetyltransferase 8 and deacetylated by histone deacetylase 7. They used LC Sciences’ m⁶A RNA sequencing service on total RNA extracted from cells and tissue samples. K235 acetylation strengthens the m⁶A demethylation activity of ALKBH5 by increasing its recognition of m⁶A on mRNA. RNA-binding protein paraspeckle component 1 (PSCP1) is a regulatory subunit of ALKBH5 and preferentially interacts with K235-acetylated ALKBH5 to recruit and facilitate the recognition of m⁶A mRNA by ALKBH5, thereby promoting m⁶A erasure. Mitogenic signals promote ALKBH5 K235 acetylation. K235 acetylation of ALKBH5 is upregulated in cancers and promotes tumorigenesis. Thus, their findings reveal that the m⁶A demethylation activity of ALKBH5 is orchestrated by its K235 acetylation and regulatory subunit PSPC1 and that K235 acetylation is necessary for the m⁶A demethylase activity and oncogenic roles of ALKBH5.

ALKBH5 acetylation at K235 is critical for the RNA m⁶A demethylation activity of ALKBH5

a, b K235 acetylation of ALKBH5 decreased the cellular mRNA m⁶A levels. The wild-type ALKBH5 and its mutant K235R and K235Q plasmids were transfected into ALKBH5 KO HeLa cells, and the cellular mRNA m⁶A level was determined by dot blotting (a) and quantified by LC‒MS/MS analysis (b) (n = 3, two-tailed unpaired Student’s t test, mean ± SD). c, d Wild-type ALKBH5, but not the K235R mutant, directly demethylated m⁶A in the m⁶A-RNA oligos in the in vitro demethylation reaction. Immunopurified wild-type ALKBH5 and its mutant K235R (c) or recombinant wild-type ALKBH5 and its mutant K235R (d) were incubated with m⁶A RNA oligos; the m⁶A level was determined by dot blotting or LC‒MS/MS assays. e Cumulative distribution curve for the m⁶A peak abundance in NC, WT and K235R cells. f Distribution of m⁶A peaks in the 5′ UTR, CDS, stop codon and 3′ UTR in NC, WT and K235R cells. g Top consensus motif identified by HOMER with m⁶A peaks in NC, WT and K235R cells. h The indicated ALKBH5 vectors together with the KAT8 plasmid were cotransfected into ALKBH5 KO HeLa cells, and the cellular RNA m⁶A level was determined. i, j Recombinant wild-type ALKBH5 or its K235R mutant was incubated with m⁶A RNA oligos after recombinant wild-type ALKBH5 or its K235R mutant was treated with immunopurified KAT8 (i) or immunopurified HDAC7 (j), and the m⁶A level was determined. Source data are provided as a Source Data file.

Zhang Xiao-Lan, Chen Xin-Hui, Xu Binwu, Chen Min, Zhu Song, Meng Nan, Wang Ji-Zhong, Zhu Huifang, Chen De, Liu Jin-Bao, Yan Guang-Rong. (2023) K235 acetylation couples with PSPC1 to regulate the m6A demethylation activity of ALKBH5 and tumorigenesis. Nature Communications 14(1), 3815. [article]

LPS adsorption and inflammation alleviation by polymyxin B-modified liposomes for atherosclerosis treatment

Admin — Mon, 26 Feb 2024 17:23:23 +0000

Chronic inflammation is critical in the onset and progression of atherosclerosis (AS). The lipopolysaccharide (LPS) level in the circulation system is elevated in AS patients and animal models, which is correlated with the severity of AS. Inspired by the underlying mechanism that LPS could drive the polarization of macrophages toward the M1 phenotype, aggravate inflammation, and ultimately contribute to the exacerbation of AS, LPS in the circulation system was supposed to be the therapeutic target for AS treatment. In the present study led by Huazhong University of Science and Technology, polymyxin (PMB) covalently conjugated to PEGylated liposomes (PLPs) were formulated to adsorb LPS through specific interactions between PMB and LPS. LC Sciences’ RNA Sequencing service was used on total RNA obtained from bone marrow- derived macrophages. In vitro, the experiments demonstrated that PLPs could adsorb LPS, reduce the polarization of macrophages to M1 phenotype and inhibit the formation of foam cells. In vivo, the study revealed that PLPs treatment reduced the serum levels of LPS and pro-inflammatory cytokines, decreased the proportion of M1-type macrophages in AS plaque, stabilized AS plaque, and downsized the plaque burdens in arteries, which eventually attenuated the progression of AS. This study highlighted LPS in the circulation system as the therapeutic target for AS and provided an alternative strategy for AS treatment.

The mechanism underlying the inhibitive effect of PLPs on LPS-induced macrophage activation

(A) Fluorescence images of macrophages treated with LPS alone or mixed with PLPs over time. The cells were stained with anti-TLR4 antibodies on the surface and counterstained with DAPI for the nucleus. Scale bar = 10 μm. Green: FITC-labeled LPS. Red: DiD-labeled PLPs. Purple: Cy3-labeled anti-TLR4 antibodies. Blue: DAPI-stained nucleus. (B) The number of differentially expressed genes (DEGs) in macrophages between different treatment groups. (C) KEGG enrichment analysis illustrating significantly enriched signaling pathways in LPS versus PBS groups based on DEGs. (D) Based on DEGs, KEGG enrichment analysis illustrates significantly enriched signaling pathways in LPS versus PLPs groups. (E–G) Heatmap of hierarchical cluster analysis displaying normalized gene expression related to Toll-like receptor signaling, NF-κB signaling, and JAK-STAT signaling by different treatments.

Liu Huiwen, Wang Honglan, Li Qiyu, Wang Yiwei, He Ying, Li Xuejing, Sun Chunyan, Ergonul Onder, Can Füsun , Pang Zhiqing, et. al. (2023) LPS adsorption and inflammation alleviation by polymyxin B-modified liposomes for atherosclerosis treatment. Acta Pharmaceutica Sinica B 13(9), 3817-3833. [article]

Protective Effect of Bilberry Anthocyanin Extracts on Dextran Sulfate Sodium-Induced Intestinal Damage in Drosophila melanogaster

Admin — Mon, 26 Feb 2024 15:49:44 +0000

Inflammatory bowel disease (IBD) is a chronic recurrent disease that can be controlled by various natural extracts. Anthocyanins (ANCs) from bilberry have significant antioxidant capacity and are widely used as food colors and antioxidants. In this study conducted by China Jiliang University, they investigated the protective effects of bilberry anthocyanin extracts (BANCs) against dextran sulphate sodium (DSS)-induced intestinal inflammation in a Drosophila melanogaster (D. melanogaster) model, and the effects on the lifespan, antioxidant capacity, intestinal characteristics, and microbiome and gene expression profiles were analyzed to elucidate the underlying biological mechanisms. In DSS-induced normal and axenic D. melanogaster, BANCs significantly increased the survival rate, maintained the intestinal morphology and integrity, and reduced the number of dead intestinal epithelial cells and the ROS level of these cells. They used LC Sciences’ 16S rRNA sequencing service to analyze the D. melanogaster intestinal microbial composition. BANC supplementation had no significant effect on the intestinal microflora of DSS-induced D. melanogaster, as demonstrated by a 16S rDNA analysis, but improved the antioxidant capacity by activating the relative gene expression of NRF2 signaling pathways in the intestine of D. melanogaster with DSS-induced inflammation. Therefore, the results demonstrate that BANCs effectively alleviate intestinal inflammatory injury induced by DSS and improve the antioxidant capacity of D. melanogaster by modulating NRF2 signaling pathways, and could thus promote the application of BANCs as functional foods.

Effects of BANCs on the DSS-induced female D. melanogaster intestinal microbiota

C, CA, D, and DA represent the blank control group, BANC control group, inflamed group, and BANCs + inflamed group, respectively. (a) Effect of BANCs on the α-diversity (Shannon and Simpson index) of the midgut microbiota of DSS-exposed D. melanogaster. The p value of all the groups was obtained by the Kruskal–Wallis test. (b) Effect of BANCs on the β-diversity of the midgut microbiota of DSS-exposed D. melanogaster and PCoA based on weighted UniFrac distance. (c) Bacterial taxonomic profiling of the four groups at the phylum level (n = 3). (d) Relative abundances of five intestinal microorganisms in DSS-induced D. melanogaster at the phylum level. The results are presented as the means ± SEMs (n = 3); statistical comparisons were performed with t tests. Statistical significance: * p < 0.05, ** p < 0.01.

Zhang G, Gu Y, Dai X. (2022) Protective Effect of Bilberry Anthocyanin Extracts on Dextran Sulfate Sodium-Induced Intestinal Damage in Drosophila melanogaster. Nutrients 14(14), 2875. [article]

Fecal microbiota and metabolomics revealed the effect of long-term consumption of gallic acid on canine lipid metabolism and gut health

Admin — Tue, 20 Feb 2024 15:52:42 +0000

Gallic acid (GA) is a natural polyphenolic compound with many health benefits. To assess the potential risk of long-term consumption of GA to gut health, healthy dogs were fed a basal diet supplemented with GA (0%, 0.02%, 0.04%, and 0.08%) for 45 d, and fecal microbiota and metabolomics were evaluated. This study, led by researchers from South China Agricultural University, demonstrated that GA supplementation regulated serum lipid metabolism by reducing serum triglyceride, fat digestibility, and Bacteroidetes/Firmicutes ratio. In addition, the relative abundance of Parasutterella was significantly lower, and the SCFAs-producing bacteria were increased along with fecal acetate and total SCFAs contents accumulation in the 0.08% GA group. They used LC Sciences’ 16S rRNA gene sequencing service on DNA extracted from dog fecal samples. Metabolomics data further elucidated that 0.08% GA significantly affected carbohydrate metabolism by downregulating succinic acid in feces, thereby alleviating inflammation and oxidative stress. Overall, this study confirmed the beneficial effects of long-term consumption of GA on lipid metabolism and gut health, and the optimal level of GA supplementation was 0.08%.

Relative abundance of fecal microbiota

Relative abundance of fecal microbiota at the (A, C) phylum and (B, D) genus levels. (E, F) LEfSe analysis between the CON and HGA groups. Different letters indicate significant (p < 0.05) difference according to ANOVA with Fisher’s LSD multiple comparison test. CON = control; LGA = 0.02% GA; MGA = 0.04% GA; HGA = 0.08% GA.

Yang K, Jian S, Guo D, Wen C, Xin Z, Zhang L, Kuang T, Wen J, Yin Y, Deng B. (2022) Fecal microbiota and metabolomics revealed the effect of long-term consumption of gallic acid on canine lipid metabolism and gut health. Food Chemistry: X 15, 100377. [article]

Intervening in hnRNPA2B1-mediated exosomal transfer of tumor-suppressive miR-184-3p for tumor microenvironment regulation and cancer therapy

Admin — Tue, 20 Feb 2024 15:05:41 +0000

Despite being a common malignant tumor, the molecular mechanism underlying the initiation and progression of triple-negative breast cancers (TNBCs) remain unclear. Tumor-associated macrophages (TAMs) are often polarized into a pro-tumor phenotype and are associated with a poor prognosis of TNBCs. Exosomes, important mediators of cell-cell communication, can be actively secreted by donor cells to reprogram recipient cells. The functions and molecular mechanisms of tumor cell-derived exosomes in TNBCs progression and TAMs reprogramming urgently need to be further explored. In this study led by Zhejiang University, they demonstrated that tumor cell-derived exosomes enriched with miR-184-3p were taken up by macrophages to inhibit JNK signaling pathway by targeting EGR1, thereby inducing M2 polarization of macrophages and synergistically promoting tumor progression. They used LC Sciences’ RNA Sequencing and miRNA Sequencing services on Total RNA that was extracted from M0 macrophages and M0-T-exo. Nanoparticles loaded with oncogene c-Myc inhibitor JQ1 could suppress the polarization process by reducing Rac1-related exosome uptake by macrophage. More importantly, it was found for the first time that tumor-suppressive miR-184-3p was actively sorted into exosomes by binding to RNA-binding protein heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1), thus facilitating tumor cell proliferation and metastasis by relieving the inhibitory effect of miR-184-3p on Mastermind-like 1 (MAML1). Overexpressing miR-184-3p in tumor cells and simultaneously knocking down hnRNPA2B1 to block its secretion through exosomes could effectively inhibit tumor growth and metastasis. Their study revealed that hnRNPA2B1-mediated exosomal transfer of tumor-suppressive miR-184-3p from breast cancer cells to macrophages was an important mediator of TNBCs progression, providing new insights into TNBCs pathogenesis and therapeutic strategies.

Exosomal miR-184-3p induced M2 macrophage polarization by inhibiting EGR1 expression and JNK signaling pathway

a Venn diagram showed the overlap of mRNAs in down-regulated genes in M0-T-exo detected by mRNA-seq and the predicted target genes of miR-184-3p. b Heatmap of 12 mRNAs screened from (a). c The expression of Egr1 gene in M0-T-exo and M0 macrophages detected by qRT-PCR. d The expression of EGR1 protein in M0-T-exo and M0 macrophages determined by IF staining. Cell nuclei were blue. Rac1 was red. e MFI of Rac1 in (d) calculated by ImageJ software. f The expression of Egr1 gene in macrophages transfected with N.C. or miR-184-3p mimics detected by qRT-PCR. g The expression of EGR1 protein in macrophages transfected with N.C. or miR-184-3p mimics determined by IF staining. Cell nuclei were blue. Rac1 was red. h MFI of Rac1 in (g) calculated by ImageJ software. Western blot assays for p-JNK expression in M0-T-exo (i) and macrophages transfected with miR-184-3p mimics (j). Data were expressed as mean ± SD (n = 3, ***p < 0.001)

Zhou Xueqing, Hong Yiling, Liu Yupeng, Wang Li, Liu Xuan, Li Yi, Yuan Hong, Hu Fuqiang. (2023) Intervening in hnRNPA2B1-mediated exosomal transfer of tumor-suppressive miR-184-3p for tumor microenvironment regulation and cancer therapy. Journal of Nanobiotechnology 21(1), 422. [article]

Humanized APOE genotypes influence lifespan independently of tau aggregation in the P301S mouse model of tauopathy

Admin — Mon, 19 Feb 2024 20:03:35 +0000

Apolipoprotein (APOE) E4 isoform is a major risk factor of Alzheimer’s disease and contributes to metabolic and neuropathological abnormalities during brain aging. To provide insights into whether APOE4 genotype is related to tau-associated neurodegeneration, this group of researchers from University of Florida have generated human P301S mutant tau transgenic mice (PS19) that carry humanized APOE alleles (APOE2, APOE3 or APOE4). In aging mice that succumbed to paralysis, PS19 mice homozygous for APOE3 had the longest lifespan when compared to APOE4 and APOE2 homozygous mice (APOE3 > APOE4 ~ APOE2). Heterozygous mice with one human APOE and one mouse Apoe allele did not show any variations in lifespan. At end-stage, PS19 mice homozygous for APOE3 and APOE4 showed equivalent levels of phosphorylated tau burden, inflammation levels and ventricular volumes. Compared to these cohorts, PS19 mice homozygous for APOE2 showed lower induction of phosphorylation on selective epitopes, though the effect sizes were small and variable. In spite of this, the APOE2 cohort showed shorter lifespan relative to APOE3 homozygous mice. None of the cohorts accumulated appreciable levels of phosphorylated tau compartmentalized in the insoluble cell fraction. They used LC Sciences’ RNA Sequencing service on total RNA extracted from frozen, pulverized mice forebrains. RNAseq analysis showed that the induction of immune gene expression was comparable across all the APOE genotypes in PS19 mice. Notably, the APOE4 homozygous mice showed additional induction of transcripts corresponding to the Alzheimer’s disease-related plaque-induced gene signature. In human Alzheimer’s disease brain tissues, they found no direct correlation between higher burden of phosphorylated tau and APOE4 genotype. As expected, there was a strong correlation between phosphorylated tau burden with amyloid deposition in APOE4-positive Alzheimer’s disease cases. Overall, their results indicate that APOE3 genotype may confer some resilience to tauopathy, while APOE4 and APOE2 may act through multiple pathways to increase the pathogenicity in the context of tauopathy.

Bulk RNAseq analysis of the forebrains of APOE homozygous mice

Differentially regulated genes from APOE2, APOE3 and APOE4 homozygous mice are presented in a heat map (A). Volcano plot of DEG and GO and KEGG pathway analysis of enriched gene sets are shown for E2H vs E3H mice (B–D), E4H vs E3H mice (E–G), and E4H vs E2H mice (H–J). Asterisks in volcano plot refer to predicted genes. The bubble plots are colored by p-value and sized by number of genes in enriched gene sets, as defined in the accompanying panel. N = 3 mice/group

Williams Tristan, Bathe Tim, Vo Quan, Sacilotto Patricia, McFarland Karen, Ruiz Alejandra Jolie, Hery Gabriela P., Sullivan Patrick, Borchelt David R., Prokop Stefan. (2023) Humanized APOE genotypes influence lifespan independently of tau aggregation in the P301S mouse model of tauopathy. Acta Neuropathologica Communications 11(1), 99. [article]

Exosomal miR-146a-5p derived from human umbilical cord mesenchymal stem cells can alleviate antiphospholipid antibody-induced trophoblast injury and placental dysfunction by regulating the TRAF6/NF-κB axis

Admin — Mon, 19 Feb 2024 17:25:57 +0000

Exosomes originating from human umbilical cord mesenchymal stem cells (hucMSC-exos) have become a novel strategy for treating various diseases owing to their ability to regulate intercellular signal communication. However, the potential of hucMSC-exos to improve placental injury in obstetric antiphospholipid syndrome and its underlying mechanism remain unclear. The objective of this study led by Shandong First Medical University was to explore the potential application of hucMSC-exos in the treatment of obstetric antiphospholipid syndrome and elucidate its underlying mechanism. In this study, hucMSC-exos ameliorated the functional impairment of trophoblasts caused by antiphospholipid antibodies in vitro and attenuated placental dysfunction in mice with obstetric antiphospholipid syndrome by delivering miR-146a-5p. They used LC Sciences’ miRNA microarray service on Total RNA that was extracted from hucMSC-exos. Exosomal miR-146a-5p suppressed the expression of tumor necrosis factor receptor-associated factor 6 (TRAF6) and inhibited the activation of NF-κB signaling, leading to the down-regulation of IL-1β and IL-18 to rescue inflammation and modulation of Cleaved-CASP3, BAX, and BCL2 to inhibit apoptosis in HTR8/SVneo cells and mice placenta. This study identified the potential molecular basis of how hucMSC-exos improved antiphospholipid antibody-induced placental injury and highlighted the functional importance of the miR-146a-5p/TRAF6 axis in the progression of obstetric antiphospholipid syndrome. More importantly, this study provided a fresh outlook on the promising use of hucMSC-exos as a novel and effective treatment approach in obstetric antiphospholipid syndrome.

MiRNA sequencing analysis of hucMSC-exos and comprehensive analysis of genes and pathways involved in hucMSC-exos treatment based on mRNA high-throughput sequencing

A Heatmap of the top 50 most-enriched miRNAs expressed in hucMSC-exos (n = 3). B Relative percentage of miRNAs in total miRNA reads. C qRT-PCR analyzed the expression levels of the top 10 most-enriched miRNAs in hucMSC–exos (n=3). D Volcano plot of RNA-seq transcriptome data displaying the pattern of the gene expression profile in the HTR8/SVneo cells with or without hucMSC-exos treatment. The aPL + EXO group was incubated with hucMSC-exos (100 ug/ml) for 24 h post aPL stimulation (200 ug/ml). Red and blue dots indicate the significantly up- or down-regulated genes, respectively. p < 0.05, |log2FC|> 1. E Representative heatmap of differentially expressed genes between the aPL + EXO group and aPL group based on the above mRNA-seq data. (Red, relatively upregulated expression; blue, relatively downregulated expression). Each column represents one individual sample, and each row represents one single gene (n = 3). F-K Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of the up- and down-regulated genes in HTR8/SVneo cells of aPL + EXO group compare with aPL group. Gene percentage refers to the percentage of genes that were significantly enriched in the corresponding secondary class. Rich factor indicates the up-regulated and down-regulated expressed genes divided by the total number of genes. The smaller the p value, the higher enrichment degree. The diameter of the dots indicate the number of genes enriched in the corresponding signal pathways

Lv Qingfeng, Wang Yuan, Tian Wei, Liu Yuqiu, Gu Mengqi, Jiang Xiaotong, Cai Yanjun, Huo Ruiheng, Li Yuchen, Li Lei. (2023) Exosomal miR-146a-5p derived from human umbilical cord mesenchymal stem cells can alleviate antiphospholipid antibody-induced trophoblast injury and placental dysfunction by regulating the TRAF6/NF-κB axis. Journal of Nanobiotechnology 21(1), 419. [article]

Extracellular vesicles derived from human ESC–MSCs target macrophage and promote anti-inflammation process, angiogenesis, and functional recovery in ACS-induced severe skeletal muscle injury

Admin — Mon, 12 Feb 2024 16:54:57 +0000

Acute compartment syndrome (ACS) is one of the most common complications of musculoskeletal injury, leading to the necrosis and demise of skeletal muscle cells. The previous study showed that embryonic stem cells-derived mesenchymal stem cells (ESC–MSCs) are novel therapeutics in ACS treatment. As extracellular vesicles (EVs) are rapidly gaining attention as cell-free therapeutics that have advantages over parental stem cells, the therapeutic potential and mechanisms of EVs from ESC–MSCs on ACS need to be explored. In this present study conducted by Zhejiang University, they examined the protective effects in the experimental ACS rat model and investigated the role of macrophages in mediating these effects. Next, they used transcriptome sequencing to explore the mechanisms by which ESC–MSC-EVs regulate macrophage polarization. Furthermore, miRNA sequencing was performed on ESC–MSC-EVs to identify miRNA candidates associated with macrophage polarization. They used LC Sciences’ microRNA sequencing service on total RNA extracted from cell samples. They found that intravenous administration of ESC–MSC-EVs, given at the time of fasciotomy, significantly promotes the anti-inflammation process, angiogenesis, and functional recovery of muscle in ACS. The beneficial effects were associated with ESC–MSC-EVs affecting macrophage polarization by delivering various miRNAs which regulate NF-κB, JAK/STAT, and PI3K/AKT pathways. Their data further illustrate that ESC–MSC-EVs mainly modulate macrophage polarization via the miR-21/PTEN, miR-320a/PTEN, miR-423/NLRP3, miR-100/mTOR, and miR-26a/TLR3 axes. Together, their results demonstrated the beneficial effects of ESC–MSC-EVs in ACS, wherein the miRNAs present in ESC–MSC-EVs regulate the polarization of macrophages.

Effects of ESC–MSC-EVs on NF-κB, JAK-STAT, and PI3K-AKT pathways

Jiang Xiangkang, Yang Jingyuan, Lin Yao, Liu Fei, Tao Jiawei, Zhang Wenbin, Xu Jiefeng, Zhang Mao. (2023) Extracellular vesicles derived from human ESC–MSCs target macrophage and promote anti-inflammation process, angiogenesis, and functional recovery in ACS-induced severe skeletal muscle injury. Stem Cell Research & Therapy 14(1), 331. [article]