Levels Of CD44 Research Articles

English

Phytoavailable cadmium (Cd) refers to the fraction of total cadmium in soil that is easily taken up by plants, which degrades the environmental quality and can cause contamination in the food chain. Cd contamination causes high carcinogenicity and toxicities, which harm both the environment and human health, including damage to the kidneys, bones, and respiratory system. In this study, seven extracting agents like ammonium bicarbonate diethylene triamine penta acetic acid (ABDTPA) and neutral salts having low and higher concentrations were used for the determination of phytoavailable Cd in sandy clay loam (SCL) and clay loam (CL) soils contaminated with Cd at naturally-Cd, Cd-20, Cd-40, and Cd-60 mg kg-1 and correlate it with Cd contents in roots, shoots, and grains of maize crop. Extraction efficiency of extractants remains in order: 10 M NH4AOc > ABDTPA > 1 M NH4AOc > 0.1 M CaCl2 > 0.01 M CaCl2 > 1 M NaNO3 > 0.1 M NaNO3. ABDTPA showed significantly the highest positive correlation with maize roots, shoots, and grains Cd contents in naturally Cd contaminated SCL and CL soils. While for artificially contaminated SCL and CL soils contaminated with Cd at Cd-20, Cd-40, and Cd-60 contamination levels, 0.1 M CaCl2 extractable Cd showed a stronger positive significant correlation with Cd contents in roots, shoots, and grains of maize crop. Overall results concluded that 10 M NH4AOc has the highest efficiency to extract Cd and 0.1 M NaNO3 has the lowest Cd extraction efficiency. Correlation results showed that ABDTPA and 0.1 M CaCl2 were found to be the most suitable extractants for predicting the bioavailability of Cd. Highly concentrated extractants give a stronger positive correlation as compared to low concentrated extractants. SCL soil showed stronger positive correlations as compared to CL soil

CD44 correlates with longevity and enhances basal ATF6 activity and ER stress resistance

The naked mole rat (NMR) is the longest-lived rodent, resistant to multiple age-related diseases including neurodegeneration. However, the mechanisms underlying the NMR's resistance to neurodegenerative diseases remain elusive. Here, we isolated oligodendrocyte progenitor cells (OPCs) from NMRs and compared their transcriptome with that of other mammals. Extracellular matrix (ECM) genes best distinguish OPCs of long- and short-lived species. Notably, expression levels of CD44, an ECM-binding protein that has been suggested to contribute to NMR longevity by mediating the effect of hyaluronan (HA), are not only high in OPCs of long-lived species but also positively correlate with longevity in multiple cell types/tissues. We found that CD44 localizes to the endoplasmic reticulum (ER) and enhances basal ATF6 activity. CD44 modifies proteome and membrane properties of the ER and enhances ER stress resistance in a manner dependent on unfolded protein response regulators without the requirement of HA. HA-independent role of CD44 in proteostasis regulation may contribute to mammalian longevity.

Activity of granulomatosis with polyangiitis and its correlation with mTOR phosphoproteomics in neutrophils.

Granulomatosis with polyangiitis (GPA) is a small vessel vasculitis with a complex pathomechanism. Organ damage in GPA is also mediated by extracellular trap formation (NETosis). We analyzed the functional status of phosphoproteins modulating NETosis in neutrophils by the mammalian target of rapamycin (mTOR) pathway in GPA along with NETosis biomarkers. Phosphoproteins levels measured in isolated neutrophils from 42 patients with GPA (exacerbation n=21; remission n=21) and 21 healthy controls were compared to serum biomarkers of the disease. Neutrophils in active disease manifested lowered levels of phosphorylated mTOR(Ser2448), PTEN(Ser380) and ULK1(Ser555), whereas phosphorylated GSK-3α/β(Ser21/Ser9) was elevated. Exacerbation of GPA was characterized by elevated neutrophil dsDNA in serum, circulating mitochondrial DNA, and DNA-MPO complexes. A significant negative correlation between mTOR or PTEN phosphoproteins and biomarkers of GPA activity was also present, reflecting the clinical activity score of GPA. Positive correlations between phosphorylated GSK-3 α/β and circulating mtDNA, DNA-MPO complexes, neutrophil-released dsDNA, or circulating proteins were also significant. Increased serum levels of IGFBP-2, TFF-3, CD147, and CHI3L1 accompanied GPA exacerbation, whereas DPP-IV levels were the lowest in active GPA. Using a principal component analysis basigin, PTEN and mTOR had the highest loadings on the discrimination function, allowing classification between active, remission, and control subjects with 98% performance. We present evidence that inhibited mTOR signaling accompanies NETosis in patients with GPA. The functional status of phosphoproteins suggests simultaneous activation of NETosis and autophagy. These results give rise to the study of autophagy as a mechanism underlying granuloma formation in GPA.

Effects of blocking CD24 and CD47 'don't eat me' signals in combination with rituximab in mantle-cell lymphoma and chronic lymphocytic leukaemia.

Mantle-cell lymphoma (MCL) is a B-cell non-Hodgkin Lymphoma (NHL) with a poor prognosis, at high risk of relapse after conventional treatment. MCL-associated tumour microenvironment (TME) is characterized by M2-like tumour-associated macrophages (TAMs), able to interact with cancer cells, providing tumour survival and resistance to immuno-chemotherapy. Likewise, monocyte-derived nurse-like cells (NLCs) present M2-like profile and provide proliferation signals to chronic lymphocytic leukaemia (CLL), a B-cell malignancy sharing with MCL some biological and phenotypic features. Antibodies against TAMs targeted CD47, a 'don't eat me' signal (DEMs) able to quench phagocytosis by TAMs within TME, with clinical effectiveness when combined with Rituximab in pretreated NHL. Recently, CD24 was found as valid DEMs in solid cancer. Since CD24 is expressed during B-cell differentiation, we investigated and identified consistent CD24 in MCL, CLL and primary human samples. Phagocytosis increased when M2-like macrophages were co-cultured with cancer cells, particularly in the case of paired DEMs blockade (i.e. anti-CD24 + anti-CD47) combined with Rituximab. Similarly, unstimulated CLL patients-derived NLCs provided increased phagocytosis when DEMs blockade occurred. Since high levels of CD24 were associated with worse survival in both MCL and CLL, anti-CD24-induced phagocytosis could be considered for future clinical use, particularly in association with other agents such as Rituximab.

Nestin, CD44, Ki-67, GS and AQP4 expression in the brain neurogenic niches of deceased patients with liver cirrhosis of different degree

The aim of the study. Immunohistochemical study of Nestin, CD44, Ki-67, GS, AQP4 expression in the subventricular zone of the lateral ventricle and hippocampus of deceased patients with liver cirrhosis depending on the age and Child–Pugh score. Materials and methods. The brains of 90 deceased patients aged 65 ± 3 years with non-alcoholic liver cirrhosis (LC) Child–Pugh class A, B and C were studied, which comprised 3 groups: group “A” – 30 deceased patients with compensated LC; “B” – 30 deceased patients with subcompensated LC (“mild decompensation”); “C” – 30 deceased patients with decompensated LC. Control group included brains of 30 patients died from acute cardiovascular failure and did not have liver disease. Each group was divided into 2 subgroups: patients ≤59 y. o. and patients ≥60 y. o. Grade 1–4 hepatic encephalopathy was detected in 59 out of 90 (65.55 %) patients with LC. The immunohistochemical levels of Nestin, CD44, Ki-67, GS and AQP4 were evaluated in paraffin tissue sections of the subventricular zones (SVZ) of the anterior and lower horns of the brain lateral ventricles, as well as the subgranular zone (SGZ) of the dentate gyrus (DG) and other structures of hippocampus in standardized fields of view of the microscope Scope A1 Carl Zeiss (Germany) using Videotest-Morphology 5.2.0.158 software. Results. In SVZ of control subgroups, Nestin+ astrocyte-like stem cells were localized mainly in subventricular glial nodules (SGN) and to a lesser extent in astrocytic ribbon. In brains of patients with compensated and subcompensated LC, there was increased Nestin expression compared to control (by 61.36 % and 208.74 %, respectively) due to increased numbers of Nestin+ cells in astrocytic ribbons. In the hippocampus of control and cirrhotic patients, Nestin expression was determined mainly in astrocyte-like cells of the fimbria-fornix, “glial plates” around the blood vessels entering the choroid plexus and subpial zone. In the SVZ of patients with subcompensated LC, the expressions of Nestin, CD44, and Ki-67 were maximally increased (by 208.74 %, 37.83 %, and 3 times, respectively), moreover, in the areas of periventricular reparative astrogliosis, in small foci of encephalolysis in the head of caudate nucleus, among GS+ and CD44+ astrocytes clusters of astrocyte-like Nestin+ and CD44+ cells were detected. In patients with decompensated LC, a significant decrease in Nestin and CD44 expression and absence of Ki-67 were observed in the SVZ, with a simultaneous maximum increase in the expression of GS and AQP4. Conclusions. In the neurogenic niches of the lateral ventricles and hippocampus of patients with compensated and subcompensated LC, there are signs of activation of neural stem cells and niche astrocytes with increased expression of Nestin, CD44, and Ki-67, which reaches maximum in subcompensated LC. Clusters of astrocyte-like Nestin+ and CD44+ cells appear in foci of periventricular repair, which probably migrate from active adjacent subventricular niche. In the brains of the patients with decompensated LC and severe Grade 3–4 hepatic encephalopathy, deep astrocytic dysmetabolic dystrophy is associated with substantial decrease in the activity of subventricular stem niche and expected astrocytogenesis.

The Effects of Premedication with Desipramine on Bone Marrow Mobilization for Autologous Bone Marrow Transplantation in Multiple Myeloma

Background: Multiple myeloma (MM) management includes primary chemotherapy, followed by autologous bone marrow transplantation (ABMT) if the bone marrow gets cleared of cancerous cells. In some cases disease relapse may occur disease relapse may occur if complete clearance is not achieved. Transplantation was primarily performed through the bone marrow; however, peripheral blood has become more favored due to its safety and convenient collection in case of appropriate bone marrow mobilization which is a challenging issue. Objectives: The current study was aimed to investigate the effectiveness of desipramine application for bone marrow mobilization in multiple myeloma (MM) who were candidate for autologous bone marrow transplantation (ABMT). Methods: The current randomized clinical trial involved 122 MM patients who were candidate for ABMT. The participants were divided into two intervention groups: the first group (n = 63) received G-CSF only treatment, while the second group (n = 59) received a similar G-CSF treatment plus desipramine. The first group received 30 microgram intravenous G-CSF for five days and the second one was treated with G-CSF with similar pattern in combination with daily 100 mg desipramine initiated within three days before G-CSF treatment and continued until the last dose of G-CSF. CD34+ cells and complete blood cells and differentiation were assessed by the end of the interventions. Results: The findings of the study show that the number of CD34+ cells, white blood cells (WBC) and platelet (PLT) count were remarkably higher among the patients receiving the combination therapy compared to the G-CSF only treated group (P-value < 0.001) within 5 days after the interventions; however, no significant differences were observed between the two groups when considering the stage of the disease and the frequency of chemotherapy sessions (P-value > 0.05). Conclusions: Desipramine application led to significantly increased levels of CD34+ cells as the representatives of bone marrow mobilization. Besides, the patients treated with this regimen had higher serum levels of WBC and PLT; however, the disease stage and the number of chemotherapy sessions did not affect the response to the treatment.

The Value of Neutrophil Gelatinase-Associated Lipocalin Receptor as a Novel Partner of CD38 in Chronic Lymphocytic Leukemia: From an Adverse Prognostic Factor to a Potential Pharmacological Target?

Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of neoplastic B lymphocytes that escape death, and correlates with the expression of negative prognostic markers such as the CD38 antigen. Although certain new drugs approved by the US Food and Drug Administration improve the clinical outcome of CLL patients, drug resistance and disease relapse still occur. Like CD38, neutrophil gelatinase-associated lipocalin receptor (NGAL-R) is frequently overexpressed in CLL cells. Here, we evaluated the concomitant surface expression of NGAL-R and CD38 in leukemic blood cells from 52 CLL patients (37 untreated, 8 in clinical remission, and 7 relapsed). We provide evidence of a positive correlation between NGAL-R and CD38 levels both in the interpatient cohorts (p < 0.0001) and in individual patients, indicating a constitutive association of NGAL-R and CD38 at the cell level. Patients with progressing CLL showed a time-dependent increase in NGAL-R/CD38 levels. In treated CLL patients who achieved clinical remission, NGAL-R/CD38 levels were decreased, and were significantly lower than in the untreated and relapsed groups (p < 0.02). As NGAL-R and CD38 participate in CLL cell survival, envisioning their simultaneous inhibition with bispecific NGAL-R/CD38 antibodies capable of inducing leukemic cell death might provide therapeutic benefit for CLL patients.

Activation of the inflammasome drives peritoneal deterioration in a mouse model of peritoneal fibrosis.

During peritoneal dialysis (PD), the peritoneum is exposed to a bioincompatible dialysate, deteriorating the tissue and limiting the long-term effectiveness of PD. Peritoneal fibrosis is triggered by chronic inflammation induced by a variety of stimuli, including peritonitis. Exposure to PD fluid alters peritoneal macrophages phenotype. Inflammasome activation triggers chronic inflammation. First, it was determined whether inflammasome activation causes peritoneal deterioration. In the in vivo experiments, the increased expression of the inflammasome components, caspase-1 activity, and concomitant overproduction of IL-1β and IL-18 were observed in a mouse model of peritoneal fibrosis. ASC-positive and F4/80-positive cells colocalized in the subperitoneal mesothelial cell layer. These macrophages expressed high CD44 levels indicating that the CD44-positive macrophages contribute to developing peritoneal deterioration. Furthermore, intravital imaging of the peritoneal microvasculature demonstrated that the circulating CD44-positive leukocytes may contribute to peritoneal fibrosis. Bone marrow transplantation in ASC-deficient mice suppressed inflammasome activation, thereby attenuating peritoneal fibrosis in a high glucose-based PD solution-injected mouse model. Our results suggest inflammasome activation in CD44-positive macrophages may be involved in developing peritoneal fibrosis. The inflammasome-derived pro-inflammatory cytokines might therefore serve as new biomarkers for developing encapsulating peritoneal sclerosis.

Granulocyte-colony stimulating factor in decompensated liver cirrhosis: a meta-analysis of four randomized controlled trials.

Decompensated liver cirrhosis (DC) has high mortality, but liver transplantation is limited due to organ scarcity and contraindications for transplantation. Granulocyte-colony stimulating factor (GCSF) shows potential for liver disease treatment with its regenerative and immunomodulatory properties. To assess the controversial use of GCSF in DC, a meta-analysis of randomized controlled trials (RCTs) compared survival benefits in patients receiving GCSF plus standard medical therapy (SMT) versus SMT alone. A literature search was performed in four databases from data inception up to December 2022, and all registered randomized controlled (RCTs) evaluating GCSF-based therapies for cirrhotic patients were included. A study combining four RCTs assessed the impact of GCSF with SMT in 595 patients with decompensated cirrhosis. The results indicated that GCSF + SMT led to higher odds of survival compared to SMT alone [risk ratio 1.28, 95% CI (1.08-1.5)]. Heterogeneity existed among the studies, but overall, GCSF showed potential in improving survival. The intervention group exhibited improved Child-Pugh-Turcotte scores [-2.51, CI (-4.33 to -0.70)], and increased CD34 levels, but no significant improvement in MELD scores. These findings suggest GCSF may benefit patients with decompensated cirrhosis in terms of survival and liver function. These results suggest that the combination of GCSF and SMT may have a positive impact on the survival rate and improvement in CPT score in patients with DC. Further RCTs are needed to shed more light on this promising modality in end-stage liver disease.

Cancer Stem Cell Markers-Clinical Relevance and Prognostic Value in High-Grade Serous Ovarian Cancer (HGSOC) Based on The Cancer Genome Atlas Analysis.

Cancer stem cells (CSCs) may contribute to an increased risk of recurrence in ovarian cancer (OC). Further research is needed to identify associations between CSC markers and OC patients' clinical outcomes with greater certainty. If they prove to be correct, in the future, the CSC markers can be used to help predict survival and indicate new therapeutic targets. This study aimed to determine the CSC markers at mRNA and protein levels and their association with clinical presentation, outcome, and risk of recurrence in HGSOC (High-Grade Serous Ovarian Cancer). TCGA (The Cancer Genome Atlas) database with 558 ovarian cancer tumor samples was used for the evaluation of 13 CSC markers (ALDH1A1, CD44, EPCAM, KIT, LGR5, NES, NOTCH3, POU5F1, PROM1, PTTG1, ROR1, SOX9, and THY1). Data on mRNA and protein levels assessed by microarray and mass spectrometry were retrieved from TCGA. Models to predict chemotherapy response and survival were built using multiple variables, including epidemiological data, expression levels, and machine learning methodology. ALDH1A1 and LGR5 mRNA expressions indicated a higher platinum sensitivity (p = 3.50 × 10-3; p = 0.01, respectively). POU5F1 mRNA expression marked platinum-resistant tumors (p = 9.43 × 10-3). CD44 and EPCAM mRNA expression correlated with longer overall survival (OS) (p = 0.043; p = 0.039, respectively). THY1 mRNA and protein levels were associated with worse OS (p = 0.019; p = 0.015, respectively). Disease-free survival (DFS) was positively affected by EPCAM (p = 0.004), LGR5 (p = 0.018), and CD44 (p = 0.012). In the multivariate model based on CSC marker expression, the high-risk group had 9.1 months longer median overall survival than the low-risk group (p < 0.001). ALDH1A1, CD44, EPCAM, LGR5, POU5F1, and THY1 levels in OC may be used as prognostic factors for the primary outcome and help predict the treatment response.

Mesenchymal stem cells shift the pro-inflammatory phenotype of neutrophils to ameliorate acute lung injury

BackgroundMesenchymal stem cell (MSC) treatment plays a major role in the management of acute lung injury (ALI), and neutrophils are the initial line of defense against ALI. However, the effect of MSCs on neutrophils in ALI remains mostly unknown.MethodsWe investigated the characteristics of neutrophils in lung tissue of ALI mice induced by lipopolysaccharide after treatment with MSCs using single-cell RNA sequencing. Neutrophils separated from lung tissue in ALI were co-cultured with MSCs, and then samples were collected for reverse transcription-polymerase chain reaction and flow cytometry.ResultsDuring inflammation, six clusters of neutrophils were identified, annotated as activated, aged, and circulatory neutrophils. Activated neutrophils had higher chemotaxis, reactive oxygen species (ROS) production, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase scores than aged neutrophils. Circulatory neutrophils occurred mainly in healthy tissue and were characterized by higher expression of Cxcr2 and Sell. Activated neutrophils tended to exhibit higher expression of Cxcl10 and Cd47, and lower expression of Cd24a, while aged neutrophils expressed a lower level of Cd47 and higher level of Cd24a. MSC treatment shifted activated neutrophils toward an aged neutrophil phenotype by upregulating the expression of CD24, thereby inhibiting inflammation by reducing chemotaxis, ROS production, and NADPH oxidase.ConclusionWe identified the immunosuppressive effects of MSCs on the subtype distribution of neutrophils and provided new insight into the therapeutic mechanism of MSC treatment in ALI.

SDPS-29 SERUM-DERIVED CD47 AS A MARKER OF TUMOR BURDENAND OUTCOME IN GBM PATIENTS – CORRELATIVE ANALYSISOF TUMOR LOCATION, RADIATION THERAPY VOLUMES,AND SURVIVAL

Abstract Glioblastomas (GBM) are the most aggressive central nervous system tumors exhibiting near universal recurrence following resection and chemoradiation (CRT). CD47 is an integrin-associated transmembrane protein associated with immunomodulation, invasion, and stemness that is highly expressed in GBM. We tested the hypothesis patient serum-derived CD47 measured pre vs. post-CRT may be related to tumor burden by exploring its association with tumor dissemination in the brain, RT volumes, and outcomes. CD47 was analyzed using an aptamer-based SOMAscan proteomic assay on serum collected pre and post CRT for 82 individuals with a pathologic diagnosis of GBM. Clinical features (tumor location (cortical/periventricular), gross tumor volumes (GTVT1, GTVT2), and outcome (overall survival (OS), progression-free survival (PFS)) were analyzed in conjunction with alteration in CD47 levels. Alteration in CD47 level was associated with OS (HR = 32.34 CI: 3.520 - 288.3, p = 0.0004053) and PFS (HR =8.5, p = .0008 CI: 0.2291-4.011). Expression of CD47 increased after CRT (p = .022). Patients with periventricular tumors had higher expression of CD47 pre vs. post CRT (p = 0.0188) and worse OS (median 13 vs. 25 months) and PFS (median 5 vs. 10 months) as compared to patients with strictly cortical tumor presence. A positive correlation was identified between GTVT1 and GTVT2 and CD47 expression levels (p = .0006). Lower CD47 expression values were associated with higher survival probability (p = .013). The stratification of CD47 expression levels yielded increasingly significant results with the separation of discrete survival groups (p = .00013), indicating clinically distinct subtypes of GBM. Tumor location (cortical vs. periventricular) and GTVT1 correlated with OS and PFS (p&amp;lt;0.05), unlike GTVT2 in this cohort in the absence of CD47 expression level. CD47 expression levels correlate with tumor location, RT volumes, OS, and PFS with distinct GBM subtypes potentially stratified via serum CD47 expression levels.

Abstract P3172: Elamipretide Reduces Mitochondrial Hyperacetylation In Left Ventricular Myocardium Of Dogs With Chronic Heart Failure

Background: The failing heart manifests abnormalities of mitochondrial (MITO) function characterized by reduced respiration and membrane potential and increased acetylation of MITO proteins. The cause(s) of MITO protein hyperacetylation is unclear, but can be due to reduced sirtuin 3 (Sirt3) possessing deacetylase activity and increased CD38 possessing NADase activity and increased NADH/NAD ratio. Long-term therapy with elamipretide (ELAM), a MITO-targeting tetra-peptide, was shown to improve MITO function in dogs with microembolization-induced heart failure (HF). Hypothesis: We tested the hypothesis that MITO hyperacetylation caused by dysregulation of Sirt3 and CD38 and increased NADH/NAD ratio in MITO of left ventricular (LV) myocardium of HF dogs can be normalized after chronic therapy with ELAM. Methods: MITO fractions were isolated from LV tissue of 14 HF dogs randomized to 3 months monotherapy with subcutaneous injections of ELAM (HF+ELAM, 0.5 mg/kg once daily, n=7) or no therapy at all (control, HF-CON, n=7) and from LV tissue of 6 normal (NL) dogs for comparison. An NAD/NADH quantitation kit was used to determined levels of NADH and NAD in MITO fractions. Protein levels of CD38, Sirt3, and porin, an internal MITO loading control, were determined in Mito-SDS-extracts using Western blotting and band intensity expressed in densitometric units (du). Protein levels of CD38 and Sirt3 were normalized to porin. Results: Protein levels of porin were similar among NL, HF-CON and HF+ELAM dogs. In HF-CON dogs, NADH/NAD ratio increased (0.70 ± 0.14 versus 0.32 ± 0.05, p&lt;0.05 versus HF) compared to NL dogs and decreased significantly in HF+ELAM (0.36 ± 0.02, p&lt;0.05 versus HF-CON). In HF-CON dogs, Sirt3 (0.50 ± 0.0.05 versus 1.24 ± 0.13, p&lt;0.05 versus NL) was significantly reduced and CD38 (1.69 ± 0.12 versus 0.70 ± 0.03, p&lt;0.05 versus NL) was significantly increased compared to NL dogs. Treatment with ELAM normalized Sirt3 (1.0 ± 0.11, p&lt;0.05 vs. HF-CON) and CD38 (0.92 ± 0.051, p&lt;0.05 vs. HF-CON). Conclusions: ELAM normalized levels of Sirt3 and CD38 and reduced NADH/NAD ratio in MITO of LV of HF dogs suggesting normalization of MITO acetylome. The later may be partly responsible for improved MITO function and possibly LV function in HF dogs treated with ELAM.

Adenosine metabolism by mesenchymal stromal cells isolated from different human tissues.

Mesenchymal stromal cells (MSCs) have unique biological properties and play important functions, which make them attractive tools for cell-based therapies. The basic mechanisms of these cells are not fully understood. However, the adenosinergic pathway contributes to the main effects attributed to MSCs. Adenosine is a highly immunosuppressive molecule and exerts a central role in inflammation by neutralizing the proinflammatory ATP influence. This nucleoside is produced by purinergic signaling, an important physiological pathway for MSCs, which involves proliferation, migration, differentiation, and apoptosis. Therefore, in this study, we analyzed the extracellular AMP hydrolysis and consequent adenosine production, as well as the expression of CD73 and adenosine receptors on the cell surface of MSCs isolated from different human tissues: dermis (D-MSCs), adipose tissue (AD-MSCs), and umbilical cord (UC-MSCs). All cells confirmed their multipotent capacity by adipogenic, osteogenic, and chondrogenic differentiation, as well as the expression of cell surface markers including CD44 + , CD105 + , and CD90 + . All MSCs expressed similar levels of CD73 and CD26 without a statistical difference among the different tissues, whereas ADA expression was lower in AD-MSCs. In addition, A1R and A3R mRNA levels were higher in D-MSCs and AD-MSCs, respectively. Enzymatic assay showed that AD-MSCs have the highest hydrolysis rate of AMP, leading to increased amount of adenosine production. Moreover, despite all MSCs completely hydrolyze extracellular AMP generating adenosine, the pattern of nucleosides metabolism was different. Therefore, although MSCs share certain characteristics as the multilineage potential and immunophenotype, they show different adenosinergic profiles according to tissue origin.

Fuyuan Huoxue Decoction Promotes Skin Flap Survival Via the P38MAPK/IKK/NF-κB Pathway in Rats

Objective Fuyuan Huoxue decoction (FHD) is a traditional Chinese medicine that can promote blood circulation, inhibit inflammatory reactions and effectively relieve pain. This study investigated the effect of FHD on rat skin flap survival and the p38 MAPK/IKK/NF-κB pathway. Methods The pathological condition of the flap tissue was examined by H&amp;E staining. The expression levels of CD34 and VEGF in skin flap tissues were examined by immunohistochemistry. The levels of TNF-α, IL-6, CRP, and HO-1, superoxide dismutase (SOD) activity, and MDA content were detected by ELISA. The protein expression levels of Bax, Bcl-2, cleaved-caspase 3, p-IKKαβ, p-p38, p-p65, α-SMA, CK14, CK15, and SOX2 were detected by western blotting. Results Fuyuan Huoxue decoction upregulated the levels of CD34, VEGF, and Bcl-2, downregulated the expression of TNF-α, IL-6, p-IKKαβ, P-P38, P-P65, Bax, and cleaved-caspase 3, increased the activity of SOD, and inhibited the level of MDA. In addition, FHD promoted the proliferation and differentiation of skin flap. Conclusions Fuyuan Huoxue decoction improved the survival of the rat skin flap model by regulating the p38MAPK/IKK/NF-κB signaling pathway, and the mechanism may be related to the promotion of skin flap proliferation, differentiation, and revascularization and the inhibition of inflammation and oxidative stress.

Gene Expression Profile of 3D Spheroids in Comparison with 2D Cell Cultures and Tissue Strains of Diffuse High-Grade Gliomas.

The use of relevant, accessible, and easily reproducible preclinical models of diffuse gliomas is a prerequisite for the development of successful therapeutic approaches to their treatment. Here we studied the gene expression profile of 3D spheroids in a comparison with 2D cell cultures and tissue strains of diffuse high-grade gliomas. Using real time PCR, we evaluated the expression of Gfap, Cd44, Pten, S100b, Vegfa, Hif1a, Sox2, Melk, Gdnf, and Mgmt genes playing an important role in the progression of gliomas and regulating tumor cell proliferation, adhesion, invasion, plasticity, apoptosis, DNA repair, and recruitment of tumor-associated cells. Gene expression analysis showed that 3D spheroids are more similar to tumor tissue strains by the expression levels of Gfap, Cd44, and Pten, while the expression levels of Hif1a and Sox2 in 3D spheroids did not differ from those of 2D cell cultures, the expression levels S100b and Vegfa in 3D spheroids was higher than in other models, and the expression levels of Melk, Gdnf, and Mgmt genes changed diversely. Thus, 3D spheroid model more closely mimics the tumor tissue than 2D cell culture, but still is not the most relevant, probably due to too small size of spheroids, which does not allow reproducing hypoxia and apoptotic and necrotic processes in the tumor tissue.

Uncovering the expression of circPVT1 in the extracellular vesicles of acute myeloid leukemia patients

Extracellular vesicles (EVs) act as molecular mediators in the tumor microenvironment, by shuttling information contained within malignant cells and functioning as regulators of the immune system. Circular (circ)RNAs are characterized by a closed loop-like structure that makes them more stable in the extracellular milieu and suitable to be packaged inside EVs. circPVT1 (hsa_circ_0001821) showed an oncogenic role in several cancer types and immunosuppressive properties in myeloid and lymphoid cell subsets. In this study, we characterized EVs from acute myeloid leukemia (AML) patients in terms of size, concentrations, surface markers and circPVT1 cargo. We showed that circPVT1 is overexpressed by primary blast cells from newly-diagnosed AML patients compared with hematopoietic stem-progenitor cells and is released as cell-free RNA in the plasma. We isolated EVs from the plasma of AML patients and healthy subjects by size exclusion chromatography and characterized them by nanoparticle tracking analysis. EVs from patients’ plasma are larger compared with those from healthy subjects and their surface profile is characterized by higher levels of the leukemic cell markers CD133, CD105, CD49e and other immune-related epitopes, with differences according to AML molecular profile. Moreover, digital PCR analysis revealed that circPVT1 is more abundant inside EVs from the plasma of AML patients compared with healthy subjects. Our findings provide new insights on the features and content of AML EVs and suggest a role of circPVT1 in the crosstalk between AML cells and the tumor microenvironment.

Role of SH3GLB1 in the regulation of CD133 expression in GBM cells

BackgroundGlioblastoma (GBM), a malignant brain tumor, has poor survival outcomes due to recurrence or drug resistance. We found that SH3GLB1 is a crucial factor for cells to evade temozolomide (TMZ) cytotoxicity through autophagy-mediated oxidative phosphorylation, which is associated with CD133 levels. Therefore, we propose that SH3GLB1 participate in the impact on tumor-initiating cells (TICs).MethodsThe parental, the derived resistant cell lines and their CD133+ cells were used, and the levels of the proteins were compared by western blotting. Then RNA interference was applied to observe the effects of the target protein on TIC-related features. Finally, in vitro transcription assays were used to validate the association between SH3GLB1 and CD133.ResultsThe CD133+ cells from resistant cells with enhanced SH3GLB1 levels more easily survived cytotoxic treatment than those from the parental cells. Inhibition of SH3GLB1 attenuated frequency and size of spheroid formation, and the levels of CD133 and histone 4 lysine 5 (H4K5) acetylation can be simultaneously regulated by SH3GLB1 modification. The H4K5 acetylation of the CD133 promoter was later suggested to be the mediating mechanism of SH3GLB1.ConclusionsThese data indicate that SH3GLB1 can regulate CD133 expression, suggesting that the protein plays a crucial role in TICs. Our findings on the effects of SH3GLB1 on the cells will help explain tumor resistance formation.

Hsa_circ_0071589 aggravates stemness and oxaliplatin resistance in colorectal cancer through sponging miR-133b to upregulate SOX13 expression.

Hsa_circ_0071589 can exacerbate the malignant behavior of colorectal cancer (CRC) cells. However, its function in stemness and oxaliplatin (OXP) resistance of CRC cells remains unclear. To assess the function of hsa_circ_0071589 in stemness and OXP resistance of CRC cells. Western blotting and qRT-PCR were applied to assess protein and mRNA levels. The association between hsa_circ_0071589, miR-133b and SOX13 was explored via a correlation analysis. Sphere formation was used to assess cell stemness. Meanwhile, the viability of CRC cells and OXP-resistant CRC cells was evaluated with the MTT assay. Cell stemness marker (CD133) levels and apoptosis of CRC cells and OXP-resistant CRC cells were tested using flow cytometry. The ALDH level was investigated using the related detection kit. In addition, the association between hsa_circ_0071589 and miR-133b and SOX13 was investigated using the RIP and dual luciferase assay. Finally, in vivo experiments were performed to detect the function of hsa_circ_0071589 in CRC, and the levels of SOX13, Ki67, and CD44 in mice were evaluated via immunohistochemistry staining. The expression of hsa_circ_0071589 and SOX13 was upregulated in CRC, whereas the expression of miR-133b was downregulated. Hsa_circ_0071589 knockdown significantly inhibited CRC stemness via the mediation of miR-133b. Moreover, hsa_circ_0071589 silencing significantly sensitized CRC cells to OXP by upregulating miR-133b. SOX13 was the direct target of miR-133b, and miR-133b could attenuate stemness and OXP resistance in CRC cells by targeting SOX13. Notably, hsa_circ_0071589 knockdown inhibited tumor growth and decreased OXP resistance in mice with CRC. Hsa_circ_0071589 aggravates stemness and OXP resistance by sponging miR-133b to indirectly target SOX13 in CRC. Thus, our study might present a novel treatment strategy against OXP-resistant CRC.

Exploring the Role of CD74 and D-Dopachrome Tautomerase in COVID-19: Insights from Transcriptomic and Serum Analyses.

The COVID-19 pandemic has posed a significant threat to public health worldwide. While some patients experience only mild symptoms or no symptoms at all, others develop severe illness, which can lead to death. The host immune response is believed to play a crucial role in determining disease severity. In this study, we investigated the involvement of CD74 and D-DT in COVID-19 patients with different disease severities, by employing an in silico analysis of a publicly available transcriptomic dataset and by measuring their serum levels by ELISA. Our results showed a significant increase in MIF levels in PBMCs from COVID-19 patients, as well as a significant increase in the D-DT levels in PBMCs. However, we observed no modulation in the serum levels of D-DT. We also observed a concordant reduction in the serum levels and PBMCs expression levels of CD74. Furthermore, we found a negative correlation between CD74 serum levels and IL-13. In conclusion, our study sheds light on the involvement of CD74 and D-DT in COVID-19, with potential implications for disease severity and treatment. Further studies are needed to fully elucidate the mechanisms underlying these observations and to explore the potential therapeutic value of targeting CD74 and IL-13 in COVID-19.