TGF-β1 Gene Expression Research Articles

Investigation of the roles of TGFβ1, CUG2, TGFBI genes, and thiol-disulfide balance on prostate cancer and metastasis

Abstract Objectives Transforming growth factor-beta (TGFβ1) is involved in tumorigenesis and metastasis. It provides this effect both by disrupting the thiol-disulfide balance and through the cancer-upregulated gene (CUG2) and transforming growth factor beta-induced (TGFBI) genes in the signaling pathway. In this study, the roles of TGFβ1 and related genes, as well as thiol-disulfide balance, in the formation of prostate cancer and metastasis were investigated. Methods Tissue samples were taken from 33 benign prostatic hyperplasia (BPH) and 35 prostate cancer (PC) patients to determine the Gleason score and metastasis. TGFβ1, CUG2, and TGFBI gene expression levels were measured by RT-PCR. Serum prostate specific antigen (PSA) levels were measured in patients, and PSA density (PSAD) was calculated. Total thiol and native thiol measurements in serum were performed spectrophotometrically, and disulfide was calculated. Results In patients with prostate cancer and metastases, PSA and PSAD levels were high, while total thiol and native thiol were significantly lower (p<0.05). TGFβ1, CUG2 and TGFBI gene expression levels were higher in patients with prostate cancer and metastases and were negatively correlated with total thiol and native thiol (p<0.001). Conclusions As a result of our study, we determined that the increase in TGFβ1, CUG 2 and TGFBI in prostate cancer plays an important role in cancer formation and metastasis by disrupting the thiol-disulfide balance.

In ovo injection of silver nanoparticles modulates some productive traits and hepatic gene expression of broilers exposed to lipopolysaccharide challenge.

This study aimed to evaluate the effectiveness of the embryonic injection of silver nanoparticles (SilNPs) on some productive traits and hepatic gene expression of lipopolysaccharide (LPS)-challenged broilers after a 42d rearing period. 560 fertile eggs were randomly allocated to four groups and received either of the following treatments at d 7 of incubation, control (no injection), placebo (1mL saline), SilNP20 (20mg/kg silver nanoparticles), or SilNP40 (40mg/kg silver nanoparticles). After the incubation, 320 hatchlings experienced a 42d standard rearing period. Live body weight (LBW), feed intake (FI), and feed conversion ratio (FCR) were weekly recorded. At the end of the experiment, two birds from each replicate (n = 8 per treatment) were exposed to LPS intraperitoneal injection at 48, 24, and 12h before slaughter time. They were also used for blood, intestinal, and microbial evaluations. The hepatic mRNA levels of tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), transforming growth factor beta (TGF-β), and insulin-like growth factor I (IGF-I) were assessed at d 1 and 42 of the experiment. Adminstration of SiLNPs improved LBW, FI, and FCR and also enhanced liver and spleen weights (P < 0.05). SilNP20 birds had significantly lower bursa of Fabricius weight (P < 0.05). SilNP20 had lower total cholesterol levels than others. There was a significant difference (P < 0.05) between SliNP40 and SilNP20 in the ratio of villus height to crypt width. Compared to control groups, chicks of SilNP20, but not SilNP40, showed a significant increase in the relative expression of TNF-α, IL-6, TGF-β, and IFG-I genes at d 1. On d 42, however, both SilNP20 and SilNP40 had significantly higher TNF-α and TGF-β levels than both controls. Silver nanoparticles did not significantly affect the microflora of the ileum and cecum in the current study. In summary, SilNPs administration to chick embryos showed a long-term positive effect on their productive performance.

The Effects of Mesenchymal Stem Cells on the Gene Expression of TGF-beta and IFN-gamma in Patients with Rheumatoid Arthritis.

The therapeutic and immunomodulatory potential of mesenchymal stem cells (MSCs) in rheumatoid arthritis (RA) has attracted considerable scientific attention in recent decades. This study aimed to evaluate the expression of genes encoding interleukin (IL)4 and IL10, as well as interferon-gamma (IFNG) and transforming growth factor beta (TGFB1) in refractory RA patients following intravenous injection of autologous bone marrow-derived MSCs (BM-MSCs). This study was registered in Iranian Registry of Clinical Trials (IRCT) (2015102824760N1) and ClinicalTrials.gov (identifier: NCT03333681). Blood samples were taken from 13 patients before and 1 and 6 months after the MSC injection to evaluate the clinical manifestations, paraclinical factors, and expression of IL4, IL10, IFNG, and TGFB1 genes employing the SYBR Green real-time reverse-transcriptase polymerase chain reaction (RT-PCR) technique. There was a significant increase in the expression of TGFB1 at 1 and 6 months after the MSC injection compared to that in the baseline, while the expression of IL4 and IL10 did not change significantly. On the other hand, the expression of IFNG increased significantly after 1 month but decreased significantly at 6 months compared to 1 month after the intervention. Nevertheless, it showed no significant decrease compared to the baseline. A significant decrease was observed for the expression of IFNG 6 months after the injection compared to that after 1 month, which was in concordance with the rise in the expression of the TGFB1 gene. A significant change in the gene expression of TGFB1 and IFNG in our study wasconsistent with the amelioration of clinical manifestations, suggesting a mechanism of action for MSCs in the treatment of RA.

Assessment of the Influence of 5-Fluorouracil on SMAD4 and TGFB1 Gene Expression, Apoptosis Induction and DNA Damage in Human Cell Lines.

Suppressor of mothers against decapentaplegic homolog 4 (SMAD family member 4, SMAD4) is involved in the adenoma-carcinoma pathway, leading to the development of colon cancer. The encoded protein is a key downstream signaling mediator in the TGFβ pathway. This pathway has tumor-suppressor functions, including cell-cycle arrest and apoptosis. Its activation in late-stage cancer can promote tumorigenesis, including metastasis and chemoresistance. Most colorectal cancer patients receive chemotherapy based on 5-FU as an adjuvant treatment. However, the success of therapy is hampered by multidrug resistance by neoplastic cells. In colorectal cancer, resistance to 5-FU-based therapy is influenced by SMAD4 gene expression, as patients with decreased SMAD4 gene expression probably have a higher risk of developing 5-FU-induced resistance. The mechanism leading to the development of this phenomenon is not fully understood. Therefore, the present study assesses the possible influence of 5-FU on changes in the expression of the SMAD4 and TGFB1 genes. The effect of 5-FU on the expression of SMAD4 and TGFB1 in colorectal cancer cells derived from the CACO-2, SW480 and SW620 cell lines was evaluated using real-time PCR. The cytotoxicity of 5-FU on colon cancer cells was assessed by the MTT method, and its effect on the induction of cell apoptosis and the initiation of DNA damage using a flow cytometer. Significant changes in the level of SMAD4 and TGFB1 gene expression were noted in the CACO-2, SW480 and SW620 cells treated with 5-FU at various concentrations during 24 h and 48 h exposure. The use of 5-FU at a concentration of 5 µmol/L resulted in a decrease in the expression of the SMAD4 gene in all cell lines at both exposure times, while the concentration of 100 µmol/L increased the expression of the SMAD4 gene in CACO-2 cells. The level of expression of the TGFB1 gene was higher for all cells treated with 5-FU at the highest concentrations, while the exposure time was extended to 48 h. The observed in vitro changes in CACO-2 cells caused by 5-FU may be of clinical relevance when choosing the drug concentration for treating patients with colorectal cancer. It is possible that 5-FU has a stronger effect on colorectal cancer cells at the higher concentrations. Low concentrations of 5-FU may not have a therapeutic effect and may also influence drug resistance in cancer cells. Higher concentrations and prolonged exposure time may affect SMAD4 gene expression, which may increase the effectiveness of therapy.

Clitoria ternatea Flower Extract-Based Gel Elevates TGF-β1 Gene Expression and Collagen Density in UVB-Induced Collagen Loss Rat Skin

Ultraviolet B (UVB) irradiation on the skin induces collagen loss by the ROS pathways and pro-collagen factors, mainly the TGF-β family. Many studies of antioxidant compounds in Clitoria ternatea have shown to induce decreasing ROS production, in the collagen loss process. However, the molecular mechanism of action is still not clearly studied, inducing the mechanism of TGF-β1 activation and regulation of collagen loss need to be explored. This study aimed to investigate the effect of Clitoria ternatea flower extract-based gel on TGF-β1 gene expression and collagen density in UVB-induced rat skin. A randomized control group post-test-only design was conducted on 20 male Wistar rats aged 8-10 weeks (150-250 grams). Rats were divided into four groups: Untreated group (Healthy Control), UVB+Based gel group (Negative Control), 5% Clitoria ternatea flower extract-based gel group (T-5%), and 10% Clitoria ternatea flower extract-based gel group (T-10%). Rats exposed to UVB for 5 consecutive days and treated with gel every day for two weeks. At week 3, rat skins were isolated for TGF-β1 gene expression using qRT-PCR, and Masson Trichrome-collagen specific staining. Comparative analysis of qRT-PCR showed that the lowest mean TGF-β1 expression was seen in the Negative Control group (0.01±0.00), followed by T-5% (0.11±0.22), Healthy Control group (1.01±0.01) and T-10% (2.32±2.46). The highest amount of collagen is in the Healthy Control group (43.83±5.3), followed by T-10% (38.39±3.1), T-5% (29.04±3.2), and the Negative Control group. (13.87±2.7). Clitoria ternatea flower extract-based gel may increase TGF-β1 gene expression and collagen deposition in the UVB-induced collagen loss rat skin.

Salivary transforming growth factor beta in oral submucous fibrosis: A diagnostic and predictive marker.

Growth factors and cytokines like transforming growth factor beta (TGF-β) play a key role in the pathogenesis of oral submucous fibrosis. To elucidate the role of Salivary TGF-β isoforms as a predictive and diagnostic marker for oral submucous fibrosis. A total of 30 OSMF and 10 control patients were included in this study, and their clinic-epidemiological data was recorded. The expression of TGF-β genes-TGF-β1, TGF-β2, TGF-β3-was studied by a real-time polymerase chain reaction in tissue and saliva. Patients were given medicinal intervention for 12 weeks along with jaw-opening exercises. Expression of salivary TGF-β genes was studied at 12 weeks. SPSS software version 20. Expression of salivary TGF beta isoforms in OSMF was more than in the control group. There was an increase in salivary TGF-β1, β2, β3 expressions with increasing clinical grades of OSMF and advancing the stage of the disease. Expression of all the TGF beta isoforms was decreased after treatment with statistically significant results. Statistically significant correlations were found between the mean difference of TGF-β1 and the mean difference between mouth opening and tongue protrusion. Salivary TGF-β isoforms may be used in diagnosis, risk assessment, and screening of the entire population at risk of OSMF after its clinical validation. However, adequate sample size and segmental assessment of the expression of TGF-β isoforms are needed for further evaluation.

In Vitro Study of the Anti-inflammatory and Antifibrotic Activity of Tannic Acid-Coated Curcumin-Loaded Nanoparticles in Human Tenocytes.

Tendinitis is a tendon disorder related to inflammation and pain, due to an injury or overuse of the tissue, which is hypocellular and hypovascular, leading to limited repair which occurs in a disorganized deposition of extracellular matrix that leads to scar formation and fibrosis, ultimately resulting in impaired tendon integrity. Current conventional treatments are limited and often ineffective, highlighting the need for new therapeutic strategies. In this work, acetalated-dextran nanoparticles (AcDEX NPs) loaded with curcumin and coated with tannic acid (TA) are developed to exploit the anti-inflammatory and anti-fibrotic properties of the two compounds. For this purpose, a microfluidic technique was used in order to obtain particles with a precise size distribution, aiming to decrease the batch-to-batch variability for possible future clinical translation. Coating with TA increased not only the stability of the nanosystem in different media but also enhanced the interaction and the cell-uptake in primary human tenocytes and KG-1 macrophages. The nanosystem exhibited good biocompatibility toward these cell types and a good release profile in an inflammatory environment. The efficacy was demonstrated by real-time quantitative polymerase chain reaction, in which the curcumin loaded in the particles showed good anti-inflammatory properties by decreasing the expression of NF-κb and TA-coated NPs showing anti-fibrotic effect, decreasing the gene expression of TGF-β. Overall, due to the loading of curcumin and TA in the AcDEX NPs, and their synergistic activity, this nanosystem has promising properties for future application in tendinitis.

Curative effect and mechanisms of Radix Arnebiae oilon burn wound healing in rats.

Radix Arnebiae oil (RAO) is a clinically useful traditional Chinese medical formula with outstanding curative effects on burns. However, the mechanism of the effect of RAO on wound healing remains unclear. The present study investigated the molecular mechanisms of the potential curative effect of RAO on wound healing. The concentrations of the main constituents shikonin, imperatorin and ferulic acid in RAO detected by HPLC were 24.57, 3.15 and 0.13 mg/mL, respectively. A rat burn model was established, and macroscopic and histopathological studies were performed. RAO significantly accelerated wound closure and repair scarring, increased superoxide dismutase activities, and reduced malondialdehyde. RAO also downregulated interleukin (IL)-6, IL-1β and tumor necrosis factor-α in wound tissues, and increased secretion of vascular endothelial growth factor, epidermal growth factor and transforming growth factor (TGF)-β1. RAO increased the gene expression of TGF-β1, type Ⅰ and Ⅲ collagen, and increased the protein expression of TGF-β1, phosphorylation of PI3K and Akt. In conclusion, RAO likely promoted wound healing via antioxidant and anti-inflammatory activities, and increased re-epithelization. Activation of the TGF-β1/PI3K/Akt pathway may play an important role in the healing efficacy of RAO. These findings suggested that RAO could be a promising alternative local treatment for burn wound healing.

Growth hormone secretagogues modulate inflammation and fibrosis in mdx mouse model of Duchenne muscular dystrophy.

Growth hormone secretagogues (GHSs) exert multiple actions, being able to activate GHS-receptor 1a, control inflammation and metabolism, to enhance GH/insulin-like growth factor-1 (IGF-1)-mediated myogenesis, and to inhibit angiotensin-converting enzyme. These mechanisms are of interest for potentially targeting multiple steps of pathogenic cascade in Duchenne muscular dystrophy (DMD). Here, we aimed to provide preclinical evidence for potential benefits of GHSs in DMD, via a multidisciplinary in vivo and ex vivo comparison in mdx mice, of two ad hoc synthesized compounds (EP80317 and JMV2894), with a wide but different profile. 4-week-old mdx mice were treated for 8 weeks with EP80317 or JMV2894 (320 µg/kg/d, s.c.). In vivo, both GHSs increased mice forelimb force (recovery score, RS towards WT: 20% for EP80317 and 32% for JMV2894 at week 8). In parallel, GHSs also reduced diaphragm (DIA) and gastrocnemius (GC) ultrasound echodensity, a fibrosis-related parameter (RS: ranging between 26% and 75%). Ex vivo, both drugs ameliorated DIA isometric force and calcium-related indices (e.g., RS: 40% for tetanic force). Histological analysis highlighted a relevant reduction of fibrosis in GC and DIA muscles of treated mice, paralleled by a decrease in gene expression of TGF-β1 and Col1a1. Also, decreased levels of pro-inflammatory genes (IL-6, CD68), accompanied by an increment in Sirt-1, PGC-1α and MEF2c expression, were observed in response to treatments, suggesting an overall improvement of myofiber metabolism. No detectable transcript levels of GHS receptor-1a, nor an increase of circulating IGF-1 were found, suggesting the presence of a novel receptor-independent mechanism in skeletal muscle. Preliminary docking studies revealed a potential binding capability of JMV2894 on metalloproteases involved in extracellular matrix remodeling and cytokine production, such as ADAMTS-5 and MMP-9, overactivated in DMD. Our results support the interest of GHSs as modulators of pathology progression in mdx mice, disclosing a direct anti-fibrotic action that may prove beneficial to contrast pathological remodeling.

Fishmeal Protein Replacement by Defatted and Full-Fat Black Soldier Fly Larvae Meal in Juvenile Turbot Diet: Effects on the Growth Performance and Intestinal Microbiota.

A 12-week feeding trial was conducted to investigate the effect of the same fishmeal protein level replaced by black soldier fly larvae (Hermetia illucens) meal (BSFL) with different lipid contents on the growth performance and intestinal health of juvenile turbot (Scophthalmus maximus L.) (initial body weight 12.64 g). Three isonitrogenous and isolipidic diets were formulated: fish meal-based diet (FM), diets DF and FF, in which 14% fish meal protein of the FM diet was replaced by defatted and full-fat BSFL, respectively. There were no significant differences in growth performance, intestinal morphology, and mucosal barrier function between the DF and the FM group. However, diet FF markedly reduced the growth performance, intestinal perimeter ratio, and the gene expression of anti-inflammatory cytokine TGF-β (P < 0.05). Compared to group FF, the communities of intestinal microbiota in group DF were more similar to group FM. Moreover, diet DF decreased the abundance of some potential pathogenic bacteria and enriched the potential probiotics, such as Bacillus. Diet FF obviously altered the composition of intestinal microbiota and increased the abundance of some potential pathogenic bacteria. These results suggested that the application of defatted BSFL showed more positive effects on fish growth and intestinal health than the full-fat BSFL, and the intestinal microbiota was closely involved in these effects.

Abstract 4919: Estrogen regulates the immune microenvironment and immunotherapy response of colorectal liver metastases

Abstract Liver metastases (LM) remain a major cause of cancer-related death and many cancers preferably metastasize to the liver due to its unique anatomical location, rich blood supply and immune-tolerant microenvironment. Up to 50% of colorectal carcinoma patients develop LM during their disease and this is associated with a poor prognosis. Our laboratory previously identified a sexual dimorphism in the regulation of the immune microenvironment (IME) of LM and showed that estrogen could promote the accumulation of myeloid-derived suppressor cells (MDSCs) and Tregs in the liver in response to invading cancer cells. The objective of this study was to elucidate the role of estrogen in the recruitment and polarization of other immune cells and to determine the therapeutic potential of Selective Estrogen Receptor Degrader (SERD) therapy in CRCLM bearing female mice, and evaluate if SERD would improve anti PD-1 immunotherapy efficacy. In estrogen-competent female mice bearing CRCLM, we found increased gene and protein expression of the immunosuppressive cytokine TGF-β, and inversely a decrease in the pro-inflammatory cytokine TNF-alpha. Furthermore, we identified a significant decrease of immunosuppressive CD68+CD163+ M2 macrophages, and an increase of the pro-inflammatory CD68+TNF-alpha+ M1 macrophages in estrogen-depleted (ovariectomized) mice as compared to estrogen-competent mice. This observation was reversed upon estradiol supplementation. Moreover, treatment of female mice with the SERD Fulvestrant, markedly reduced the number of CRCLM compared to vehicle-treated mice, reduced the cell frequencies and counts of M2 macrophages, significantly increased NK cells recruitment in the liver. The addition of SERD to immunotherapy has a better therapeutic effect than both therapies alone, and increased CD8+ T-cells, NK cells, and M1/M2 macrophages ratio. Taken together, our results identify estrogen as a critical regulator of an immunosuppressive TME in the liver and a potential therapeutic target to enhance immunotherapy efficacy. Citation Format: Yasmine Benslimane, Sarah Lapin, Julien Chambon, Stephanie Perrino, Pnina Brodt. Estrogen regulates the immune microenvironment and immunotherapy response of colorectal liver metastases. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4919.

Chamomile and Urtica dioica extracts improve immunological and histological alterations associated with polycystic ovarian syndrome in DHEA -induced mice

BackgroundOne of the novel mechanisms in the pathogenesis of Polycystic ovary syndrome (PCOS) is low-grade chronic inflammation. Chamomile (Matricaria recutita L.) and Nettle (Urtica dioica), with phytoestrogenic and antioxidant properties, are traditionally used to treat gynecological diseases. This study investigated the immune-modulating effects of these two plants.MethodsFollowing the induction of PCOS by subcutaneous injection (SC) of Dehydroepiandrosterone (DHEA) in BALB / C mice. Mice were treated in five groups: Sham, PCOS, PCOS + Chamomile, PCOS + Nettle, and PCOS + Chamomile and Nettle for 21 days. Ovarian morphology, blood antioxidant capacity, the abundance of Treg cells, and expression of matrix metalloproteinase-9 (MMP-9), transforming growth factor-ß (TGF-ß), cyclooxygenase-2 genes (COX-2), and tumor necrosis factor-alpha (TNF-α) were measured.ResultsFolliculogenesis, Cystic follicles, and corpus luteum improved in the treatment groups (P < 0. 05). Treg cells in the DHEA group were significantly reduced compared to the Sham group (P < 0. 01). However, this decrease was not corrected in treatment groups (P > 0. 05). Total serum antioxidant capacity was significantly increased in the treatment group of Nettle and Chamomile + Nettle (P < 0. 05). The expression of MMP9 and TGFβ genes in the PCOS group was significantly higher than the Sham group (P < 0. 05), which the expression of MMP9 was corrected by treatment with Chamomile + Nettle extract (P < 0. 05).ConclusionChamomile and Nettle extract may be an effective supplement in improving the histological and immunological changes of PCOS. However, more research is needed to confirm its effectiveness in humans.

Tyndallized Bacteria Preferentially Induce Human Macrophage M1 Polarization: An Effect Useful to Balance Allergic Immune Responses and to Control Infections

Macrophage polarization is a dynamic process through which macrophages acquire specific features whose extremes are represented by M1 and M2 polarization. Interleukin (IL)-6, IL-1β, IL-12 and IL-8 belong to M1 macrophages while transforming growth factor-beta (TGF-β belongs to M2 cytokines. M2 polarization prevalence is observed in allergic diseases. Tyndallization is a thermal process able to inactivate microorganisms and to allow their use for chronic respiratory disease treatment via immune response modulation. The present study explores the effects of a blend of tyndallized bacteria (TB) on macrophage polarization. THP-1-derived macrophages were exposed to different concentrations of TB (106, 5 × 106, 107, 5 × 107, 108 CFU/mL) and then cell viability and TB phagocytosis, and IL-8, IL-1β, IL-6, IL-12 and TGF-β1 gene expression and release were assessed. TB were tolerated, phagocyted and able to increase IL-8, IL-1β and IL-6 gene expression and release IL-12 gene expression, as well as decrease TGF-β1 gene expression and release. The effects on IL-8, IL-6 and TGF-β1 release were confirmed in human monocyte-derived macrophages (hMDMs) exposed to TB. In conclusion, TB promote M1 polarization, and this mechanism might have valuable potential in controlling allergic diseases and infections, possibly preventing disease exacerbations.

Differential optineurin expression controls TGFβ signaling and is a key determinant for metastasis of triple negative breast cancer.

Triple-negative breast cancer (TNBC) is the most challenging breast cancer subtype to treat due to its aggressive characteristics and low response to the existing clinical therapies. Distant metastasis is the main cause of death of TNBC patients. Better understanding of the mechanisms underlying TNBC metastasis may lead to new strategies of early diagnosis and more efficient treatment. In our study, we uncovered that the autophagy receptor optineurin (OPTN) plays an unexpected role in TNBC metastasis. Data mining of publicly available data bases revealed that the mRNA level of OPTN in TNBC patients positively correlates with relapse free and distance metastasis free survival. Importantly, in vitro and in vivo models demonstrated that OPTN suppresses TNBC metastasis. Mechanistically, OPTN inhibited the pro-oncogenic transforming growth factor-β (TGFβ) signaling in TNBC cells by interacting with TGFβ type I receptor (TβRI) and promoting its ubiquitination for degradation. Consistent with our experimental findings, the clinical TNBC samples displayed a negative correlation between OPTN mRNA expression and TGFβ gene response signature and expression of proto-typic TGFβ target genes. Altogether, our study demonstrates that OPTN is a negative regulator for TGFβ receptor/SMAD signaling and suppresses metastasis in TNBC.

Nasal administration of anti-CD3 mAb (Foralumab) downregulates NKG7 and increases TGFB1 and GIMAP7 expression in T cells in subjects with COVID-19

T cells are present in early stages of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and play a major role in disease outcome and long-lasting immunity. Nasal administration of a fully human anti-CD3 monoclonal antibody (Foralumab) reduced lung inflammation as well as serum IL-6 and C-reactive protein in moderate cases of COVID-19. Using serum proteomics and RNA-sequencing, we investigated the immune changes in patients treated with nasal Foralumab. In a randomized trial, mild to moderate COVID-19 outpatients received nasal Foralumab (100 μg/d) given for 10 consecutive days and were compared to patients that did not receive Foralumab. We found that naïve-like T cells were increased in Foralumab-treated subjects and NGK7+ effector T cells were reduced. CCL5, IL32, CST7, GZMH, GZMB, GZMA, PRF1, and CCL4 gene expression were downregulated in T cells and CASP1 was downregulated in T cells, monocytes, and B cells in subjects treated with Foralumab. In addition to the downregulation of effector features, an increase in TGFB1 gene expression in cell types with known effector function was observed in Foralumab-treated subjects. We also found increased expression of GTP-binding gene GIMAP7 in subjects treated with Foralumab. Rho/ROCK1, a downstream pathway of GTPases signaling was downregulated in Foralumab-treated individuals. TGFB1, GIMAP7, and NKG7 transcriptomic changes observed in Foralumab-treated COVID-19 subjects were also observed in healthy volunteers, MS subjects, and mice treated with nasal anti-CD3. Our findings demonstrate that nasal Foralumab modulates the inflammatory response in COVID-19 and provides a novel avenue to treat the disease.

Profiling of Immunomodulatory Genes and Quantification of CD25+ Cells in Different Types of Early Pregnancy Loss.

Maternal regulatory T (Treg) cells play a pivotal role in establishing general immune homeostasis in the decidua for maintenance of pregnancy. We aimed in this study to investigate the relationship between mRNA expression of immunomodulatory genes and CD25+ Treg cells with early pregnancy losses. Our study included 3 groups of early pregnancy losses including sporadic spontaneous abortions, recurrent spontaneous abortions, sporadic spontaneous abortions post IVF treatment and the control group. We performed RT-PCR for analyzing mRNA expression levels of 6 immunomodulatory genes and CD25 immunohistochemistry for quantification of Treg cells. Only FOXP3, CD274 (PDL1), and TGFβ1 mRNA expression levels were significantly decreased in the miscarriage groups in comparison to the control group, whereas there was no significant mRNA expression change of CD4, IL2RA, and IL10. We also found significantly lower number of CD25+ cells in the miscarriages. We conclude that decreased expression of FOXP3 and PD-L1 may play a significant role in the pathogenesis of spontaneous abortion cases whereas decreased expression of TGFβ1 gene may be associated with the occurrence of early loss in IVF-treated pregnancies. Additional immunoprofiling of Treg cell population is needed to quantify Treg cells in early pregnancy losses.

MicroRNA-122 mimic/microRNA-221 inhibitor combination as a novel therapeutic tool against hepatocellular carcinoma.

Therapeutic microRNAs (miRNAs) delivery holds a lot of promise for treating human malignancies. So, this study was carried out to examine the potential of miR-122 mimic and/or miR-221 inhibitor as an innovative therapeutic strategy for HCC in an animal model. Mice were categorized into five groups comprising: (1) a normal control group, (2) an HCC group subjected to diethylnitrosamine (DEN) injection for 12 weeks, (3) a miR-122 mimic-treated HCC group, (4) a miR-221 inhibitor-treated HCC group, and (5) a miR-122 mimic/miR-221 inhibitor-treated HCC group. After 16 weeks, all animals were sacrificed and underwent biochemical, miRNAs and genes expression, histopathological, and immunohistochemical examinations. The miR-122 mimic/miR-221 inhibitor combination dramatically reduced the levels of pro-inflammatory, liver cancer, angiogenesis, and cell proliferation markers when compared to either treatment alone. It also down-regulated the expression of cyclin D1, TGF-β, and β-catenin genes, which are involved in promoting cell cycle progression and cancer cell proliferation. Furthermore, it caused the resolution of nearly all the histological malignant features as well as the reduction of malignant cellular markers, including α-smooth muscle actin, arginase-1, and tropomyosin-1. The co-treatment with miR-122 mimic and miR-221 inhibitor amplifies the benefits of either treatment on HCC through targeting the SENP1 and ARF4 genes, respectively. This combination can inhibit cancer cell proliferation and angiogenesis while inducing tumor apoptosis and necrosis. This study demonstrates the therapeutic potential of reversing a dysregulated miRNAs expression pattern in HCC. As a result, future research should concentrate on turning miRNA understanding into therapeutic applications.

Xenogeneic mesenchymal stem cell biocurative improves skin wounds healing in diabetic mice by increasing mast cells and the regenerative profile.

Diabetes mellitus (DM) is a chronic disease and a major cause of mortality and morbidity worldwide. The hyperglycemia caused by DM induces micro and macrovascular complications that lead, among other consequences, to chronic wounds and amputations. Cell therapy and tissue engineering constitute recent therapeutic alternatives to improve wound healing in diabetic patients. The current study aimed to analyze the effectiveness of biocuratives containing human mesenchymal stem cells (MSCs) associated with a hydrogel matrix in the wound healing process and related inflammatory cell profile in diabetic mice. Biocuratives containing MSCs were constructed by 3D bioprinting, and applied to skin wounds on the back of streptozotocin (STZ)-induced type 1 diabetic (T1D) mice. The healing process, after the application of biocuratives with or without MSCs was histologically analyzed. In parallel, genes related to growth factors, mast cells (MC), M1 and M2 macrophage profiles were evaluated by RT-PCR. Macrophages were characterized by flow cytometry, and MC by toluidine blue staining and flow cytometry. Mice with T1D exhibited fewer skin MC and delayed wound healing when compared to the non-diabetic group. Treatment with the biocuratives containing MSCs accelerated wound healing and improved skin collagen deposition in diabetic mice. Increased TGF-β gene expression and M2 macrophage-related markers were also detected in skin of diabetic mice that received MSCs-containing biocuratives. Finally, MSCs upregulated IL-33 gene expression and augmented the number of MC in the skin of diabetic mice. These results reveal the therapeutic potential of biocuratives containing MSCs in the healing of skin wounds in diabetic mice, providing a scientific base for future treatments in diabetic patients.

TGFβ carrying exosomes in plasma: potential biomarkers of cancer progression in patients with head and neck squamous cell carcinoma.

Contributions of TGFβ to cancer progression are well documented. However, plasma TGFβ levels often do not correlate with clinicopathological data. We examine the role of TGFβ carried in exosomes isolated from murine and human plasma as a contributor to disease progression in head and neck squamous cell carcinoma (HNSCC). The 4-nitroquinoline-1-oxide (4-NQO) mouse model was used to study changes in TGFβ expression levels during oral carcinogenesis. In human HNSCC, TGFβ and Smad3 protein expression levels and TGFB1 gene expression were determined. Soluble TGFβ levels were evaluated by ELISA and TGFβ bioassays. Exosomes were isolated from plasma using size exclusion chromatography, and TGFβ content was quantified using bioassays and bioprinted microarrays. During 4-NQO carcinogenesis, TGFβ levels in tumour tissues and in serum increased as the tumour progressed. The TGFβ content of circulating exosomes also increased. In HNSCC patients, TGFβ, Smad3 and TGFB1 were overexpressed in tumour tissues and correlated with increased soluble TGFβ levels. Neither TGFβ expression in tumours nor levels of soluble TGFβ correlated with clinicopathological data or survival. Only exosome-associated TGFβ reflected tumour progression and correlated with tumour size. Circulating TGFβ+ exosomes in the plasma of patients with HNSCC emerge as potential non-invasive biomarkers of disease progression in HNSCC.

Effects of Six Weeks High-Intensity Interval Training on TGF-β1 and SMAD7 Genes Expression in the Kidney Tissue of Elderly Diabetic Male Wistar Rats

Introduction: Diabetic nephropathy is a common complication in the patients with diabetes. The aim of this study was to investigate the effect of six weeks of high-intensity interval training on TGF-b1 and SMAD7 gene expression in kidney tissue of elderly diabetic male Wistar rats.&#x0D; Methods: In this study, 36 male 21-month old Wistar rats, with the weight of 340 to 390 grams were randomly divided into three groups: healthy control, diabetes, and diabetes- training. The training group, performed high-intensity interval training five times per week for six weeks, healthy and diabetic control groups have not done any exercises. 24 hours after the last training session, rats were sacrificed and kidney tissues were extracted to examine TGF-b1 and SMAD7 genes expression by Real-Time PCR method. To determine the difference among the groups, one-way analysis of variance with Tukeys Post hoc test was used at the significant level of P&lt;0.05. Statistical analysis was done by SPSS software version 16.&#x0D; Results: The results showed that six weeks of high intensity interval training led to a significant decrease in TGF-b1 gene expression in kidney tissue compared to the diabetic group (P=0.001). In addition, a significant increase in SMAD7 gene expression in kidney tissue was observed in the diabetes- training group compared to the diabetes group (P=0.008).&#x0D; Conclusion: According to the findings of the present study, it seems that high-intensity interval training can probably exert protective effects against nephropathy by reducing the expression of the TGF-b1gene and increasing the expression of the SMAD7 gene in kidney tissue. Because this research was conducted on elderly diabetic rats, it is suggested to check and confirm this training method in diabetic human models.