MUC1 Levels Research Articles

Functionalized carbon nanotube field-effect transistor biosensor for highly sensitive detection of exosomal protein

Since the exosomal protein level is related to many diseases, sensitive detection of exosomal protein is highly desirable. Here, we describe a polymer-sorted high-purity semiconducting carbon nanotubes (CNTs) films-based field-effect transistor (FET) biosensor for ultrasensitive and label-free detection of MUC1, a transmembrane protein highly expressed in breast cancer exosomes. Polymer-sorted semiconducting CNTs hold advantages including high purity (>99%), high CNT concentration, and short processing time (<1 h), but they are difficult to be stably functionalized with biomolecules because of lacking hanging bonds on their surface. To solve this issue, poly-lysine (PLL) was employed to modify the CNT films after they were deposited on the sensing channel surface of the fabricated FET chip. To specifically recognize the exosomal protein, sulfhydryl aptamer probes were immobilized on the gold nanoparticles (AuNPs) surface that was assembled on PLL substrate. The aptamer-modified CNT FET was capable of sensitively and selectively detecting exosomal MUC1 as high as 0.34 fg/mL. Moreover, the CNT FET biosensor was able to recognize breast cancer patients from healthy individuals by comparing the expression level of exosomal MUC1. The developed CNT FET biosensor is expected to be a novel assay for early diagnosis of cancer.

Influenza virus infection modulates expression of the anti-inflammatory mucin 1 in both epithelial and immune cells

Abstract Cell-associated mucin 1(MUC1/Muc1) is a component of the mucus barrier in the respiratory tract that acts as a physical barrier and signaling molecule. Our prior work shows that MUC1 expression is increased during influenza virus (IAV) infection in human airway epithelial (HAE) cultures in vitro. Here we further detail MUC1 expression patterns and report that IAV-induced cytokines, including type I interferons (IFNs), trigger the expression of a low-molecular-weight form of MUC1 which localizes to the cytoplasm and nucleus, and is expressed in different epithelial cell types. Further, IFN-driven changes in the cellular composition of HAE toward a more secretory phenotype require MUC1, suggesting that MUC1 plays an important role in shaping the epithelial response during viral infection. Using a mouse model of IAV infection, we confirmed the upregulation of Muc1 throughout the epithelium in vivoand observed altered Muc1 levels in lung immune cells. Specifically, Muc1 was upregulated in interstitial macrophages but downregulated in antigen-presenting cells (APCs) including alveolar macrophages and dendritic cells as early as 2 days post-infection (dpi). By 5 dpi, Muc1 was significantly upregulated in various T cell subsets and their effector counterparts including CD4 +, CD8 +, and double-negative effector T cells. We also found a negative correlation between Muc1 levels in APCs and the expansion of effector T cell populations, supporting the putative immunoregulatory roles of MUC1/Muc1. Our ongoing work further explores the impact of Muc1 expression in specific cellular subsets on the immune response and IAV pathogenesis. This work was supported by the National Heart, Lung, and Blood Institute (R01HL151840, to MAS)

A Review of Salivary Biomarkers in Breast Cancer

Objective: This study aimed to review the literature related to salivary biomarkers used to diagnose breast cancer or predict responses to therapy and its prognosis. Methods: Studies were eligible for inclusion if they had compared salivary diagnostics for breast cancer with the gold standard or other biomarkers. Six databases (PubMed, EMBASE, ScienceDirect, Scopus, Web of Science, and Ovid) and Google Scholar search engine searched for pertinent literature using specific search terms: breast cancer, diagnosis, prognosis saliva, and salivary biomarker. Of the 513 studies screened, 110 were selected for inclusion in this review. Different salivary biomarkers were classified and discussed. Results: Our analysis indicated that the sensitivity and specificity of salivary biomarkers, such as cerb- B2 and sialic acid in BC detection are comparable with serum values. Salivary levels of autoantibodies against MUC1, CA 15-3, and adiponectin had a better correlation with breast cancer than serum levels. Assessment of biomarkers, such as HER-2, helps develop a treatment plan and evaluates response to treatment. Conclusion: Our study confirms that salivary biomarkers are becoming increasingly important in breast cancer diagnosis and treatment planning, as well as in predicting therapeutic response and prognosis. Salivary biomarkers such as c-erb-B2 in association with traditional criteria offer the promise of use as a noninvasive screening method for breast cancer. Additional studies are warranted to determine the most sensitive and specific salivary biomarkers for this purpose.

Comparison of mucin levels at the ocular surface of visual display terminal users with and without dry eye disease

BackgroundThe long-term use of visual display terminals (VDT) is linked to an increased risk of dry eye disease (DED). Numerous studies have indicated that ocular mucins play a vital role in the pathogenesis of DED. Therefore, we aimed to evaluate (1) whether mRNA levels of membrane-associated mucins (MAMs), including MUC1, MUC4, MUC16, and MUC20, as well as MUC5AC are altered in conjunctival cells of VDT users with and without DED and (2) the relationship between mucin levels and subjective and objective tests of DED in VDT users.MethodsSeventy-nine VDT users were enrolled and divided into DED (n = 53) and control (n = 26) groups. All participants were evaluated for parameters of DED using the Ocular Surface Disease Index (OSDI) questionnaire, tear breakup time (TBUT), corneal fluorescein staining (CFS), lissamine green (LG) staining, and tear meniscus height (TMH). Based on the conjunctival impression cytology (CIC) method, differences in MUC1, MUC4, MUC16, MUC20, and MUC5AC mRNA expression levels were observed between the DED and control groups, and between symptomatic and asymptomatic participants.ResultsThe DED group showed significantly decreased MUC1, MUC16, and MUC20 expressions (all P < 0.05) compared to the control group. In addition, these mucin levels were lower in subjects with frequent ocular symptoms (foreign body sensation, blurred vision and painful or sore eyes) than in asymptomatic participants (all P < 0.05). Correlation analysis revealed that MUC1, MUC16, and MUC20 levels in VDT users were positively correlated with TBUT or TMH, or both. However, no significant relationship was found between MUC4 and MUC5AC levels and the DED parameters.ConclusionVDT users with an increased frequency of ocular discomfort or a diagnosis of DED had a decreased MUC1, MUC16 and MUC20 mRNA expression in their conjunctival cells. MAMs deficiency in the conjunctival epithelium may be one of the mechanisms leading to tear film instability and DED in VDT users.

Abstract LB214: A first-in-class bispecific antibody-drug conjugate (DM002) targeting HER3 and the juxtamembrane domain of MUC1

Abstract Despite the entry of new anti-cancer drugs into the market, there were more than 10 million deaths from cancer globally in 2021, with cancers of the lung, stomach, breast, and pancreas contributing to the most cancer-related deaths in China and the USA. Accordingly, there is an urgent need for improved therapeutic interventions. Antibody-drug conjugates (ADC) are novel drugs that exploit the specificity of a monoclonal antibody for target antigens expressed on cancer cells in order to achieve targeted delivery of a potent cytotoxic payload. More recently, bispecific ADCs (BsADC) targeting two tumor-associated antigens (TAA) have been developed to further amplify tumor cell specificity while minimizing toxicity to normal tissue, thus allowing for broader therapeutic windows. HER3 and MUC1 are two TAAs that are commonly expressed/co-expressed on multiple tumor types, including lung, gastric, breast and pancreatic cancer; however, neither antigen has been successfully targeted by effective drugs: of the candidates targeting HER3 antigen being evaluated in human clinical studies, most are well-tolerated, but with limited efficacy. On the other hand, high levels of MUC1 in diseased tissues can undergo auto-proteolysis, so that drugs targeting the MUC1-N region will be neutralized before reaching the tumor tissues, thereby limiting the recognition of tumor cells. To overcome these challenges, we generated fully human bispecific antibody candidates with human-monkey cross-species reactivity that target HER3 and the juxtamembrane domain of MUC1. These bsAbs exhibit higher endocytic activity in HER3-low tumor cell lines compared with other currently available HER3 mAb. These bsAbs were subsequently conjugated with monomethyl auristatin E (MMAE) via a protease-cleavable linker to obtain first-in-class BsADC candidates, DM002. DM002 candidates showed robust anti-tumor activity in multiple CDX and PDX models of lung, breast, gastric and pancreatic cancer; most notably, DM002 candidates outperformed benchmark ADCs in BP0508 lung PDX models. Together, these data indicate that DM002 will be a promising therapeutic drug for patients with HER3 and MUC1 co-expressing tumors. Citation Format: Yifu Zhang, Chenzhang Shang, Nannan Wang, Gao An, Ellen Zhang, Qingcong Lin, Yi Yang. A first-in-class bispecific antibody-drug conjugate (DM002) targeting HER3 and the juxtamembrane domain of MUC1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB214.

Adhesion to the Brain Endothelium Selects Breast Cancer Cells with Brain Metastasis Potential.

Tumor cells metastasize from a primary lesion to distant organs mainly through hematogenous dissemination, in which tumor cell re-adhesion to the endothelium is essential before extravasating into the target site. We thus hypothesize that tumor cells with the ability to adhere to the endothelium of a specific organ exhibit enhanced metastatic tropism to this target organ. This study tested this hypothesis and developed an in vitro model to mimic the adhesion between tumor cells and brain endothelium under fluid shear stress, which selected a subpopulation of tumor cells with enhanced adhesion strength. The selected cells up-regulated the genes related to brain metastasis and exhibited an enhanced ability to transmigrate through the blood-brain barrier. In the soft microenvironments that mimicked brain tissue, these cells had elevated adhesion and survival ability. Further, tumor cells selected by brain endothelium adhesion expressed higher levels of MUC1, VCAM1, and VLA-4, which were relevant to breast cancer brain metastasis. In summary, this study provides the first piece of evidence to support that the adhesion of circulating tumor cells to the brain endothelium selects the cells with enhanced brain metastasis potential.

Abstract 1148: 1XX mutations slow CAR T cell signaling and increase in vivo persistence

Abstract Purpose: Develop CAR T cells that have increased persistence in vivo and kill low antigen expressing solid tumor cancer cells. Experimental: Three CARs were generated that were all targeted to the tumor with the same antibody fragment, huMNC2, that recognizes a cryptic epitope on the MUC1 cleavage product called MUC1* (muk 1 star). One construct comprises a 41BB co-stimulatory domain, while the second construct has a CD28 co-stimulatory domain. The first 2 CARs have a wild-type CD3z, whereas the 3rd bears Michel Sadelain’s “1XX” mutations in CD3z. These are 2 Tyrosine to Phenylalanine mutations in ITAM 2 and ITAM 3. The 1XX mutations were designed to slow signaling to eliminate the problem of CAR T cell exhaustion. Human T cells were separately transduced with the 3 CARs and tested in NSG mice bearing human breast tumors. The tumors were heterogeneous tumors comprising parent cells that fluoresce red and express medium to low amounts of the antigen, MUC1*, and cells that fluoresce green wherein the parent cells have been transduced to express more MUC1*. The percent of the tumor that expressed high levels of MUC1* was varied from 7.5% to 15% to 30%. 150 female NSG mice were implanted with 500,000 tumor cells. The animals received a single CAR T cell injection of one of the three CAR T cells at an Effector to Target ratio of 10:1, 5:1, or 1:1. The experiment was allowed to proceed for 96 days, with periodic IVIS measurements to track tumor growth with a focus on tumor recurrence. After sacrifice, residual tumors were excised, dissociated and analyzed to determine if tumor recurrence was being driven by the high antigen cells or the low antigen cells. CAR T cells were recovered from the blood and the spleen. They were enumerated and analyzed by flow cytometry for the presence of markers of exhaustion. Unpublished Results: Tumor recurrence occurred in mice treated with CAR T cells bearing wild-type CD3z, but essentially did not recur in the mice treated with CAR T cells bearing the 1XX mutations. Four hundredfold more CAR T cells were recovered from the mice treated with the CAR-1XX cells compared to CARs with wild-type CD3z, and the CAR-1XX cells appeared not to be exhausted even after 96 days. CAR-1XX T cells administered at a 1:1 E:T ratio, with only 7.5% of the tumor comprised of high antigen expressing cells, were effectively killed with no significant tumor recurrence. In contrast, the CARs with wild-type CD3z had no effect on tumor growth in this group. Surprisingly, analysis of excised tumors showed that tumor recurrence in mice treated with CAR wild-type CD3z T cells was driven by the low antigen density cells, whereas the CAR-1XX T cells killed high antigen and low antigen density cells at the same rate. Conclusions: 1XX mutations in CD3z greatly increase CAR T persistence in vivo and unexpectedly enable the killing of low antigen tumor cells. Citation Format: Cynthia C. Bamdad, Benoit J. Smagghe, Mark G. Carter, Danica M. Walkley, Jac-Leen S. Nash, Kevin R. Yi, Trevor J. Grant, Laura M. Reale, Michael J. Nash, Gregory L. Riley, Andrew K. Stewart. 1XX mutations slow CAR T cell signaling and increase in vivo persistence [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 1148.

Transcriptomic profile of the porcine oviductal epithelial cells during the peri-conceptional period.

The optimal environment in the oviduct is created by adjusting its ultrastructure and secretory activity to serve the most suitable protection of gametes and to support embryo development. Through gametes/embryo's presence inside the oviduct, the oviductal transcriptomic profile may be altered, and these changes may be caused by DNA methylation. The results of the present study documented that in the epithelial cells of the ampulla and isthmus of the oviducts collected from pigs during the peri-conceptional period, the most differentially expressed genes (DEGs) were down-regulated. Identified DEGs were classified into gene ontology categories as well as annotated into different biological pathways. From evaluated DEGs, genes important for embryo development were selected and the level of DNA methylation was determined. It was documented CLDN18, MUC1, CYP19A3, SOCS1, and ESR1 methylation level have been altered. The presence of embryos in the oviduct changed the transcriptomic profile and the level of DNA methylation in the epithelial cells of ampulla and isthmus during the peri-conceptional period.

Prevention of Inflammation-Driven Colon Carcinogenesis in Human MUC1 Transgenic Mice by Vaccination with MUC1 DNA and Dendritic Cells

The preventive efficacy of MUC1-specific DNA immunization on inflammation-driven colon carcinogenesis in human MUC1 transgenic (MUC1.Tg) mice was investigated. Mice were vaccinated with MUC1 DNA mixed with autologous bone-marrow-derived dendritic cells (BMDCs), and then colonic tumors were induced by azoxymethane (AOM) injection and oral administration of dextran sulfate sodium (DSS). Two types of tumors, squamous metaplasia and tubular adenoma, were observed. Both expressed high levels of MUC1 as indicated by the binding of anti-MUC1 antibodies with different specificities, whereas MUC1 expression was not detected in normal colonic mucosa. When mice were immunized with MUC1 DNA + BMDCs, tumor incidence, tumor number, and tumor size were significantly reduced. In contrast, vaccination with MUC1 DNA alone or BMDCs alone was ineffective in reducing tumor burden. Inflammation caused by DSS was not suppressed by the MUC1 DNA + BMDCs vaccination. Furthermore, MUC1 protein expression levels, as judged by anti-MUC1 antibody binding in tumors grown after vaccination, did not significantly differ from the control. In conclusion, an inflammation-driven carcinogenesis model was established in MUC1.Tg mice, closely resembling human colon carcinogenesis. In this model, vaccination with MUC1 DNA + BMDCs was effective in overriding MUC1 tolerance and reducing the tumor burden by a mechanism not affecting the level of colonic inflammation.

Comparative efficacy of a novel Bacillus subtilis-based probiotic and pharmacological zinc oxide on growth performance and gut responses in nursery pigs

In this study, we assessed the efficacy of a novel Bacillus subtilis probiotic in improving growth performance and gut responses in comparison to pharmacological zinc oxide (ZnO) in nursery pigs. A total of 96 piglets were randomly assigned to four groups: Negative control (NC), Positive control (PC, 3000 mg Zn /kg feed), B.subtilis low dose (BS9-L, 2 × 107 CFU/pig) and B.subtilis high dose (BS9-H, 2 × 109 CFU/pig). Growth performance, diarrhea rate, gut mucosal gene expression and fecal microbial populations were evaluated. B.subtilis administration did not improve piglet bodyweight. BS9-L showed (P < 0.05) higher average daily gain (ADG) in Period 2 (D14-D28). BS9 groups had (P < 0.001) lower feed conversion ratio (FCR) in Period 2 (D14-D28) and overall. Like the ZnO-group, BS9 groups had lower (P < 0.01) diarrhea rate. A significant reduction (P < 0.05) in fecal E. coli, total coliforms, and an increase in lactic acid bacteria and Bacillus spp. in BS9 groups was observed. BS9 group had reduced (P < 0.05) mRNA levels of intestinal IL-8 and higher levels of MUC-1 and occludin and TJP-1 compared to negative control. These findings suggest that probiotic BS9, may promote growth performance, and ameliorate various indicators of intestinal health in piglets. Hence, it may serve as a prospective alternative to ZnO growth promoter in commercial swine production.

Downregulation of Mucin1 in Cancer Cells is Associated with Modulation of Calcium Signalling Pathways and Alteration in Procoagulant Related Activity

Background Intracellular calcium ions play an essential role in regulating numerous physiological functions by acting as a second messenger in signal transduction pathways. Alterations in calcium signaling are also implicated in several pathological conditions including cancer. Aim The purpose of this study is to investigate whether MUC1, a well-established tumor marker, plays a role in modulating calcium signalling in cancer cells. Methods MUC1 knockdown breast cancer MCF-7 cells were used to investigate whether the overexpression of MUC1 by cancer cells has an influence on calcium signalling pathways, by determining the level of a range of calcium signalling proteins and the dynamics of intracellular calcium mobilization as well as the cellular effects induced by calcium treatment. The investigation was also extended to normal human mammary epithelial, human breast and pancreatic cancer cell lines, which express different levels of MUC1. Result MUC1 downregulation altered the aberrant expression of several calcium transporter proteins in MCF-7 cancer cells. The level of each of these calcium signaling pathways proteins was correlated to MUC1 level in normal human mammary epithelial, human breast and pancreatic cancer cell lines. Consequently, a number of proteins whose activities are calcium dependent such as p-CaMKII, and pro-coagulant proteins Va, Xa and thrombin were found to be reduced in MUC1 downregulated MCF-7 cells. In addition, the effect of thrombin on calcium mobilization and the cleavage of the thrombin receptors was also reduced in MUC1 downregulated MCF-7 cells. Conclusion The downregulation of MUC1 is capable of modulating calcium cellular effects via alteration in the expression of calcium signalling proteins and by impacting calcium induced initiation of procoagulant activity, which is capable of influencing the effect of thrombin on cancer cells. Such properties contribute to another crucial role for MUC1 within the signalling network associated with tumorigenesis.

MUC5AC and MUC5B expression in canine gallbladder mucocele epithelial cells.

Gallbladder mucocele (GBM) is one of the most common gallbladder diseases in dogs. Its pathogenesis has not yet been clarified, but excessive accumulation of a secretory gel-forming mucin, MUC5AC in the gallbladder has been reported. This study aimed to ascertain if MUC5AC overproduction resulted in mucus accumulation in the gallbladder during GBM development. Eleven dogs undergoing cholecystectomy who were pathologically diagnosed with GBM were included, and the expression level of mucins, particularly MUC5AC and MUC5B, in their gallbladder epithelial cells was compared with those in normal gallbladder epithelial cells. On reverse transcription-quantitative polymerase chain reaction screening, there was a significant difference (P<0.05) in the mRNA expression level of MUC1, but not of other mucins including MUC5AC and MUC5B, between mucocele and normal gallbladder epithelial cells. Protein expression levels were also evaluated for MUC5AC and MUC5B using immunohistochemistry. There was little immunoreactivity for MUC5AC, whereas MUC5B showed definitive staining in gallbladder epithelial cells. There was no difference in MUC5AC and MUC5B protein expression levels between mucocele and normal gallbladder epithelial cells. These data suggest that excessive production of mucin, especially MUC5AC and MUC5B, does not occur in canine GBM, and that abnormal mucus excretion, rather than excessive mucus production, may be the cause of GBM development.

TUMOR MARKERS EXPRESSION LEVELS IN GASTRIC CANCER PATIENT'S PERIPHERAL BLOOD BY RT-PCR ASSESSMENT.

Hematological recurrence is the second most frequent cause of failure in the treatment of gastric cancer. The detection of circulating tumor markers in peripheral blood by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) method may be a useful tool to predict recurrence and determine the patient's prognosis. However, no consensus has been reached regarding the association between the tumor markers level in peripheral blood and its impact on patient survival. To evaluate the expression of the circulating tumor markers CK20 and MUC1 in peripheral blood samples from patients with gastric cancer by qRT-PCR, and to verify the association of their expression levels with clinicopathological characteristics and survival. A total of 31 patients with gastric adenocarcinoma were prospectively included in this study. CK20 and MUC1 expression levels were analyzed from peripheral blood by the qRT-PCR technique. There was no statistically significant (p>0.05) association between CK20 expression levels and clinical, pathological, and surgical features. Higher MUC1 expression levels were associated with female patients (p=0.01). There was a correlation between both gene levels (R=0.81, p<0.001), and CK20 level and tumor size (R=0.39, p=0.034). CK20 and MUC1 expression levels could be assessed by qRT-PCR from total peripheral blood samples of patients with gastric cancer. CK20 levels were correlated to MUC1 levels as well as to tumor size. There was no difference in disease-free survival and overall survival regarding both genetic markers expression in this series.

Transcorneal but not transpalpebral electrical stimulation disrupts mucin homeostasis of the ocular surface

PurposeTranscorneal electrical stimulation (TcES) is increasingly applied as a therapy for preserving and improving vision in retinal neurodegenerative and ischemic disorders. However, a common complaint about TcES is its induction of eye pain and dryness in the clinic, while the mechanisms remain unknown.MethodTcES or transpalpebral ES (TpES) was conducted in C57BL6j mice for 14 days. The contralateral eyes were used as non-stimulated controls. Levels of intracellular [Ca2+] ([Ca2+]i) were assessed by Fura-2AM. The conductance resistances of the eye under various ES conditions were measured in vivo by an oscilloscope.ResultsAlthough TcES did not affect tear production, it significantly induced damage to the ocular surface, as revealed by corneal fluorescein staining that was accompanied by significantly decreased mucin (MUC) 4 expression compared to the control. Similar effects of ES were detected in cultured primary corneal epithelium cells, showing decreased MUC4 and ZO-1 levels after the ES in vitro. In addition, TcES decreased secretion of MUC5AC from the conjunctiva in vivo, which was also corroborated in goblet cell cultures, where ES significantly attenuated carbachol-induced [Ca2+]i increase. In contrast to TcES, transpalpebral ES (TpES) did not induce corneal fluorescein staining while significantly increasing tear production. Importantly, the conductive resistance from orbital skin to the TpES was significantly smaller than that from the cornea to the retina in TcES.ConclusionTcES, but not TpES, induces corneal epithelial damage in mice by disrupting mucin homeostasis. TpES thus may represent a safer and more effective ES approach for treating retinal neurodegeneration clinically.

The ulcerative colitis-associated gene FUT8 regulates the quantity and quality of secreted mucins

Mucins are the main macrocomponents of the mucus layer that protects the digestive tract from pathogens. Fucosylation of mucins increases mucus viscoelasticity and its resistance to shear stress. These properties are altered in patients with ulcerative colitis (UC), which is marked by a chronic inflammation of the distal part of the colon. Here, we show that levels of Fucosyltransferase 8 (FUT8) and specific mucins are increased in the distal inflamed colon of UC patients. Recapitulating this FUT8 overexpression in mucin-producing HT29-18N2 colonic cell line increases delivery of MUC1 to the plasma membrane and extracellular release of MUC2 and MUC5AC. Mucins secreted by FUT8 overexpressing cells are more resistant to removal from the cell surface than mucins secreted by FUT8-depleted cells (FUT8 KD). FUT8 KD causes intracellular accumulation of MUC1 and alters the ratio of secreted MUC2 to MUC5AC. These data fit well with the Fut8-/- mice phenotype, which are protected from UC. Fut8-/- mice exhibit a thinner proximal colon mucus layer with an altered ratio of neutral to acidic mucins. Together, our data reveal that FUT8 modifies the biophysical properties of mucus by controlling levels of cell surface MUC1 and quantity and quality of secreted MUC2 and MUC5AC. We suggest that these changes in mucus viscoelasticity likely facilitate bacterial-epithelial interactions leading to inflammation and UC progression.

Defects in the expression of colonic host defense factors associate with barrier dysfunction induced by a high-fat/high-cholesterol diet.

The effect of Western diets in the gastrointestinal system is largely mediated by their ability to promote alterations in the immunity and physiology of the intestinal epithelium, and to affect the composition of the commensal microbiota. To investigate the response of the colonic epithelium to high-fat/high-cholesterol diets (HFHCDs), we evaluated the synthesis of host defense factors involved in the maintenance of the colonic homeostasis. C57BL/6 mice were fed an HFHCD for 3 weeks and their colons were evaluated for histopathology, gene expression, and microbiota composition. In addition, intestinal permeability and susceptibility to Citrobacter rodentium were also studied. HFHCD caused colonic hyperplasia, loss of goblet cells, thinning of the mucus layer, moderate changes in the composition of the intestinal microbiota, and an increase in intestinal permeability. Gene expression analyses revealed significant drops in the transcript levels of Muc1, Muc2, Agr2, Atoh1, Spdef, Ang4, Camp, Tff3, Dmbt1, Fcgbp, Saa3, and Retnlb. The goblet cell granules of HFHCD-fed mice were devoid of Relmβ and Tff3, indicating defective production of those two factors critical for intestinal epithelial defense and homeostasis. In correspondence with these defects, colonic bacteria were in close contact with, and invading the epithelium. Fecal shedding of C. rodentium showed an increased bacterial burden in HFHCD-fed animals accompanied by increased epithelial damage. Collectively, our results show that HFHCD perturbs the synthesis of colonic host defense factors, which associate with alterations in the commensal microbiota, the integrity of the intestinal barrier, and the host's susceptibility to enteric infections.

Tumour-associated Mucin1 correlates with the procoagulant properties of cancer cells of epithelial origin

BackgroundCancer-associated thrombosis (CAT) is caused, at least in part, by procoagulant factors produced by the tumour itself. Although MUC1 is an established biomarker for the diagnosis, immunotherapy, and prognosis of cancer, it is unclear whether it contributes to the procoagulant phenotype of cancer cells. MethodsMUC1 knockdown breast cancer MCF-7 cells were used to investigate the influence of overexpression of MUC1 on procoagulant parameters. In addition, the effect of treating normal human epithelial cells with extracellular vesicles from several human breast and pancreatic cancer cell lines, which overexpress MUC1, was determined. The impact of a pharmacological anti-MUC1 antibody on cancer cells was also analysed. ResultsThe level of a range of procoagulant proteins was observed to correlate with the MUC1 level of human breast and pancreatic cancer cell lines. MUC1 downregulation in MCF-7 cells led to a reduction in the procoagulant parameters particularly thrombin activity. The levels of selected tumorigenic markers, procoagulant proteins and miRNAs associated with tumorigenicity and thromboembolism were also modulated by treatment of normal cells with tumour cell derived extracellular vesicles in correlation with that of the extracellular vesicles donor cells. Moreover, the procoagulant properties were also reduced by an anti-MUC1 antibody in these cancer cells. ConclusionsA range of procoagulant proteins found in human breast and pancreatic cancer cells were shown to exhibit a positive correlation with the level of MUC1 and thereby potentially contribute to the pathogenesis of CAT. The reduction of the procoagulant activity by MUC1 antibody could be an additional beneficial effect of its therapeutic efficacy. These findings also suggest that the level of tumour associated MUC1 could be of use as a risk factor for CAT.

Mucin expression in liver tumors

Difficulties in diagnosis, the prevalence of oncological processes, their medical, social and economic significance and high mortality rate determine the relevance of the fight against them and make the problem of studying the mechanisms of their early diagnosis one of the key ones. The subject of research now is the description of many biological markers, the determination of which allows us to assess the risks of developing a disease, diagnose a tumor, and serve as a biological indicator of the tumor process. The subjects of the study were tissue samples from 65 patients with liver cancer. The level of antibodies to MUC-1 and MUC-13 was studied. The established value of the concentration of antibodies to MUC-1 and MUC-13 in the blood serum of people with liver cancer is higher than the values in practically healthy individuals (p = 0.002; p = 0.0001), and the concentration of antibodies to MUC-1 and MUC-13 in the tissue tumors of cancer patients is higher than in the blood serum of healthy individuals and patients with liver cancer (p&lt;0.05). The expression level of MUC-1 and MUC-13 in the tumor tissue is not related to the age and sex of patients.

Overexpression of Decay Accelerating Factor Mitigates Fibrotic Responses to Lung Injury.

CD55 or decay accelerating factor (DAF), a ubiquitously expressed glycosylphosphatidylinositol (GPI)-anchored protein, confers a protective threshold against complement dysregulation which is linked to the pathogenesis of idiopathic pulmonary fibrosis (IPF). Since lung fibrosis is associated with downregulation of DAF, we hypothesize that overexpression of DAF in fibrosed lungs will limit fibrotic injury by restraining complement dysregulation. Normal primary human alveolar type II epithelial cells (AECs) exposed to exogenous complement 3a or 5a, and primary AECs purified from IPF lungs demonstrated decreased membrane-bound DAF expression with concurrent increase in the endoplasmic reticulum (ER) stress protein, ATF6. Increased loss of extracellular cleaved DAF fragments was detected in normal human AECs exposed to complement 3a or 5a, and in lungs of IPF patients. C3a-induced ATF6 expression and DAF loss was inhibited using pertussis toxin (an enzymatic inactivator of G-protein coupled receptors), in murine AECs. Treatment with soluble DAF abrogated tunicamycin-induced C3a secretion and ER stress (ATF6 and BiP expression) and restored epithelial cadherin. Bleomycin-injured fibrotic mice subjected to lentiviral overexpression of DAF demonstrated diminished levels of local collagen deposition and complement activation. Further analyses showed diminished release of DAF fragments, as well as reduction in apoptosis (TUNEL and caspase 3/7 activity), and ER stress-related transcripts. Loss-of-function studies using Daf1 siRNA demonstrated worsened lung fibrosis detected by higher mRNA levels of Col1a1 and epithelial injury-related Muc1 and Snai1, with exacerbated local deposition of C5b-9. Our studies provide a rationale for rescuing fibrotic lungs via DAF induction that will restrain complement dysregulation and lung injury.

Effects of dietary dihydromyricetin supplementation on intestinal barrier and humoral immunity in growing-finishing pigs

In this study, we investigated the effect of dietary dihydromyricetin (DHM) supplementation on intestinal barrier and humoral immunity in growing-finishing pigs. The data showed that dietary DHM supplementation improved jejunal barrier function by upregulating the protein expressions of Occludin and Claudin-1 and the mRNA levels of MUC1 and MUC2. Dietary DHM supplementation increased the amylase, lipase, sucrase and maltase activities and the mRNA expression of nutrient transporter (SGLT1, GLUT2, PepT1) in the jejunum mucosa. Dietary DHM supplementation significantly reduced the E. coli population in the cecum and colon and increased the Lactobacillus population in the cecum. In addition, dietary DHM supplementation increased the contents of butyric acid and valeric acid in cecum and colon. In serum, dietary DHM supplementation reduced interleukin-1β (IL-1β) content and increased interleukin-10 (IL-10), Immunoglobulin M (IgM) and Immunoglobulin A (IgA) contents (p < 0.05). In addition, compared with the control group, dietary DHM supplementation improved secretory immunoglobulin A (sIgA) and interleukin-10 (IL-10) contents and down-regulated TNF-α protein expression in jejunum mucosa (p < 0.05). Together, this study demonstrated that dietary DHM supplementation improved intestinal barrier function, digestion and absorption capacity and immune function in growing-finishing pigs.