Search

Gluten is a key component that allows wheat flour to form a dough, and it is also a byproduct of the production of wheat starch. As a commercial product, wheat gluten is increasingly used in the food-related industry because of its versatile functional properties and wide range of sources. Wheat gluten is manufactured industrially on a large scale through the Martin process and batter process and variants thereof. Gliadin and glutenin impart cohesiveness and elasticity properties, respectively, to wheat gluten. The formation of gluten networks and polymers depends mainly on covalent bonds (disulfide bonds) and noncovalent bonds (ionic bonds, hydrogen bonds, and hydrophobic interactions). The multifunctional properties (viscoelasticity, gelation, foamability, etc.) of wheat gluten are shown by rehydration and other processing techniques. Wheat gluten has been widely used in wheat-based products, food auxiliary agents, food packaging, encapsulation and release of food functional ingredients, food adsorption and heat insulation materials, special purpose foods, and versatile applications. In the future, wheat gluten protein will be used as an important raw material to participate in the development and preparation of various food and degradable materials, and the application potential of wheat gluten in food-related industries will be massive. This review summarizes the main manufacturing processes, composition, and structure of gluten protein, and the various functional properties that support its application in the food and related industries.The versatile functional properties of wheat gluten are closely related to structureThe application as a binder and substitute for meat is growing in popularityRenewable and degradable gluten-based materials are important exploited fieldsWheat gluten shows vast potential and application value in food-related industry.

Purpose:All uveal melanoma and a fraction of other melanoma subtypes are driven by activation of the G-protein alpha-q (Gαq) pathway. Targeting these melanomas has proven difficult despite advances in the molecular understanding of key driver signaling pathways in the disease pathogenesis. Inhibitors of Gαq have shown promising preclinical results, but their therapeutic activity in distinct Gαq mutational contexts and in vivo have remained elusive.Experimental Design:We used an isogenic melanocytic cellular system to systematically examine hotspot mutations in GNAQ (e.g., G48V, R183Q, Q209L) and CYSLTR2 (L129Q) found in human uveal melanoma. This cellular system and human uveal melanoma cell lines were used in vitro and in in vivo xenograft studies to assess the efficacy of Gαq inhibition as a single agent and in combination with MEK inhibition.Results:We demonstrate that the Gαq inhibitor YM-254890 inhibited downstream signaling and in vitro growth in all mutants. In vivo, YM-254890 slowed tumor growth but did not cause regression in human uveal melanoma xenografts. Through comprehensive transcriptome analysis, we observed that YM-254890 caused inhibition of the MAPK signaling with evidence of rebound by 24 hours and combination treatment of YM-254890 and a MEK inhibitor led to sustained MAPK inhibition. We further demonstrated that the combination caused synergistic growth inhibition in vitro and tumor shrinkage in vivo.Conclusions:These data suggest that the combination of Gαq and MEK inhibition provides a promising therapeutic strategy and improved therapeutic window of broadly targeting Gαq in uveal melanoma.See related commentary by Neelature Sriramareddy and Smalley, p. 1217

Supplementary figures S1-S6 with figure legends.

raw uncropped labeled western blots

Purpose: All uveal melanoma and a fraction of other melanoma subtypes are driven by activation of the G-protein alpha-q (Gαq) pathway. Targeting these melanomas has proven difficult despite advances in the molecular understanding of key driver signaling pathways in the disease pathogenesis. Inhibitors of Gαq have shown promising preclinical results, but their therapeutic activity in distinct Gαq mutational contexts and in vivo have remained elusive. Experimental Design: We used an isogenic melanocytic cellular system to systematically examine hotspot mutations in GNAQ (e.g., G48V, R183Q, Q209L) and CYSLTR2 (L129Q) found in human uveal melanoma. This cellular system and human uveal melanoma cell lines were used in vitro and in in vivo xenograft studies to assess the efficacy of Gαq inhibition as a single agent and in combination with MEK inhibition. Results: We demonstrate that the Gαq inhibitor YM-254890 inhibited downstream signaling and in vitro growth in all mutants. In vivo, YM-254890 slowed tumor growth but did not cause regression in human uveal melanoma xenografts. Through comprehensive transcriptome analysis, we observed that YM-254890 caused inhibition of the MAPK signaling with evidence of rebound by 24 hours and combination treatment of YM-254890 and a MEK inhibitor led to sustained MAPK inhibition. We further demonstrated that the combination caused synergistic growth inhibition in vitro and tumor shrinkage in vivo. Conclusions: These data suggest that the combination of Gαq and MEK inhibition provides a promising therapeutic strategy and improved therapeutic window of broadly targeting Gαq in uveal melanoma. See related commentary by Neelature Sriramareddy and Smalley, p. 1217

BackgroundDynamic N6-methyladenosine (m6A) RNA modification generated and erased by N6-methyltransferases and demethylases regulates gene expression, alternative splicing and cell fate. Ocular melanoma, comprising uveal melanoma (UM) and conjunctival melanoma (CM), is the most common primary eye tumor in adults and the 2nd most common melanoma. However, the functional role of m6A modification in ocular melanoma remains unclear.Methodsm6A assays and survival analysis were used to explore decreased global m6A levels, indicating a late stage of ocular melanoma and a poor prognosis. Multiomic analysis of miCLIP-seq, RNA-seq and Label-free MS data revealed that m6A RNA modification posttranscriptionally promoted HINT2 expression. RNA immunoprecipitation (RIP)-qPCR and dual luciferase assays revealed thatHINT2mRNA specifically interacted with YTHDF1. Furthermore, polysome profiling analysis indicated a greater amount ofHINT2mRNA in the translation pool in ocular melanoma cells with higher m6A methylation.ResultsHere, we show that RNA methylation significantly inhibits the progression of UM and CM. Ocular melanoma samples showed decreased m6A levels, indicating a poor prognosis. Changes in global m6A modification were highly associated with tumor progression in vitro and in vivo. Mechanistically, YTHDF1 promoted the translation of methylatedHINT2mRNA, a tumor suppressor in ocular melanoma.ConclusionsOur work uncovers a critical function for m6A methylation in ocular melanoma and provides additional insight into the understanding of m6A modification.

To understand the formation of gluten network and its regulation on noodle qualities upon mixing and resting, the dynamic distribution and molecular transformation of gluten were tracked and quantified. Confocal laser scanning microscopy and scanning electron microscopy images showed that appropriate mixing (8 min) and resting (60 min) induced a compact gluten network with higher gluten junctions. Both height and width of protein molecular chains were increased by hydration during mixing and reduced after excessive resting (90 min). According to the size exclusion/reversed phase-HPLC profiles, mixing induced slight depolymerization of large glutenin polymer, and α-gliadin subunits were more susceptible to polymerization after appropriate mixing and resting. Increased mixing time was accompanied by the strengthening of ionic and hydrogen bonds, and the weakening of hydrophobic interaction. PCA and correlation analysis revealed the accurate regulation of mixing and resting induced dynamic distribution and evolution of gluten on the macroscopic noodle qualities.

N

Uveal melanoma (UM) and retinoblastoma (RB), which cause blindness and even death, are the most frequently observed primary intraocular malignancies in adults and children, respectively. Epigenetic studies have shown that changes in the epigenome contribute to the rapid progression of both UM and RB following classic genetic changes. The loss of epigenetic homeostasis plays an important role in oncogenesis by disrupting the normal patterns of gene expression. The targetable nature of epigenetic modifications provides a unique opportunity to optimize treatment paradigms and establish new therapeutic options for both UM and RB with these aberrant epigenetic modifications. We aimed to review the research findings regarding relevant epigenetic changes in UM and RB. Herein, we 1) summarize the literature, with an emphasis on epigenetic alterations, including DNA methylation, histone modifications, RNA modifications, noncoding RNAs and an abnormal chromosomal architecture; 2) elaborate on the regulatory role of epigenetic modifications in biological processes during tumorigenesis; and 3) propose promising therapeutic candidates for epigenetic targets and update the list of epigenetic drugs for the treatment of UM and RB. In summary, we endeavour to depict the epigenetic landscape of primary intraocular malignancy tumorigenesis and provide potential epigenetic targets in the treatment of these tumours.

Chromatin remodeling impacts the structural neighborhoods and regulates gene expression. However, the role of enhancer-guided chromatin remodeling in the gene regulation remains unclear. Here, using RNA-seq and ChIP-seq, we identified for the first time that neurotensin (NTS) serves as a key oncogene in uveal melanoma and that CTCF interacts with the upstream enhancer of NTS and orchestrates an 800 kb chromosomal loop between the promoter and enhancer. Intriguingly, this novel CTCF-guided chromatin loop was ubiquitous in a cohort of tumor patients. In addition, a disruption in this chromosomal interaction prevented the histone acetyltransferase EP300 from embedding in the promoter of NTS and resulted in NTS silencing. Most importantly, in vitro and in vivo experiments showed that the ability of tumor formation was significantly suppressed via deletion of the enhancer by CRISPR-Cas9. These studies delineate a novel onco-enhancer guided epigenetic mechanism and provide a promising therapeutic concept for disease therapy.

Most uveal melanomas harbor mutations in Gαq and show constitutive MAPK activation. Although MEK inhibition has some efficacy against uveal melanoma, clinical responses are typically poor. The Gαq inhibitor-MEK inhibitor combination showed prolonged suppression of MAPK signaling in preclinical uveal melanoma models and led to improved therapeutic responses.

Aberrant chromatin transformation dysregulates gene expression and may be an important driver of tumorigenesis. However, the functional role of chromosomal dynamics in tumorigenesis remains to be elucidated. Here, using in vitro and in vivo experiments, we reveal a novel long noncoding (lncing) driver at chr12p13.3, in which a novel lncRNA GALNT8 Antisense Upstream 1 (GAU1) is initially activated by an open chromatin status, triggering recruitment of the transcription elongation factor TCEA1 at the oncogene GALNT8 promoter and cis-activates the expression of GALNT8. Analysis of The Cancer Genome Atlas (TCGA) clinical database revealed that the GAU1/GALNT8 driver serves as an important indicative biomarker, and targeted silencing of GAU1 via the HKP-encapsulated method exhibited therapeutic efficacy in orthotopic xenografts. Our study presents a novel oncogenetic mechanism in which aberrant tuning of the chromatin state at specific chromosomal loci exposes factor-binding sites, leading to recruitment of trans-factor and activation of oncogenetic driver, thereby provide a novel alternative concept of chromatin dynamics in tumorigenesis.

Background Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a hereditary disease caused by a mutation in the forkhead box L2 ( FOXL2 ) gene. Female patients suffering from premature ovarian failure (POF) were classified as type I, and others were classified as type II. We aimed to clarify a novel FOXL2 indel mutation in Chinese families and to predict the POF risk in the affected patient.Methods Three generations of one Chinese family with BPES were enrolled in this study. Blood samples from patients of this family were collected and then analysed by whole-exome sequencing. Confocal microscopy was performed to observe the subcellular location. Transactivation studies were performed with real-time PCR.Results This novel mutation (c.1068G>C) is located in the downstream of DNA-binding forkhead(FHD) domain, and the mutant protein could also exhibit transactivation capacity of StAR , a key regulator of POF. Conclusively, we discovered a novel FOXL2 mutation and predicted that female patient in this family should be classified as type II BPES.Conclusions Our study revealed a novel missense mutation (c.1068G>C) and expanded the spectrum of FOXL2 gene mutations. Although we were not able to determine the classification from clinical manifestation, we discovered the patient developed type II BPES through subcellular distribution and transactivation analysis.

Background Dynamic N6-methyladenosine (m6A) RNA modification generated and erased by N6-methyltransferases and demethylases regulates gene expression, alternative splicing and cell fate. Ocular melanoma, comprising uveal melanoma (UM) and conjunctival melanoma (CM), is the most common primary eye tumor in adults and the 2nd most common melanoma. However, the functional role of m6A modification in ocular melanoma remains unclear. Methods m6A assays and survival analysis were used to explore decreased global m6A levels, indicating a late stage of ocular melanoma and a poor prognosis. Multiomic analysis of miCLIP-seq, RNA-seq and Label-free MS data revealed that m6A RNA modification posttranscriptionally promoted HINT2 expression. RNA immunoprecipitation (RIP)-qPCR and dual luciferase assays revealed that HINT2 mRNA specifically interacted with YTHDF1. Furthermore, polysome profiling analysis indicated a greater amount of HINT2 mRNA in the translation pool in ocular melanoma cells with higher m6A methylation. Results Here, we show that RNA methylation significantly inhibits the progression of UM and CM. Ocular melanoma samples showed decreased m6A levels, indicating a poor prognosis. Changes in global m6A modification were highly associated with tumor progression in vitro and in vivo. Mechanistically, YTHDF1 promoted the translation of methylated HINT2 mRNA, a tumor suppressor in ocular melanoma. Conclusions Our work uncovers a critical function for m6A methylation in ocular melanoma and provides additional insight into the understanding of m6A modification.

Additional file 12: Table S5. Oligonucleotides used in this study.

N6-methyladenosine (m6A), the most prevalent modification of mammalian RNA, has received increasing attention. Although m6A has been shown to be associated with biological activities, such as spermatogenesis modulation, cell spermatogenesis and pluripotency, Drosophila sex determination, and the control of T cell homeostasis and response to heat shock, little is known about its roles in cancer biology and cancer stem cells. Recent articles have noted that some genes have abnormal m6A expression after tumorigenesis, including genes ABS2, RARA, MYB, MYC, ADAM19 and FOX1. Abnormal changes in the m6A levels of these genes are closely related to tumour occurrence and development. In this review, we summarized the ‘dual edge weapon’ role of RNA methylation in the tumorigenesis. We discussed RNA methylation could lead to not only tumour progression but also tumour suppression. Moreover, we clarified that the abnormal changes in the m6A enrichment of specific loci contribute to tumour occurrence and development, thereby representing a novel anti-cancer strategy by restoration to balanced RNA methylation in tumour cells.

Exploring gene function is critical for understanding the complexity of life. DNA sequences and the three-dimensional organization of chromatin (chromosomal interactions) are considered enigmatic factors underlying gene function, and interactions between two distant fragments can regulate transactivation activity via mediator proteins. Thus, a series of chromosome conformation capture techniques have been developed, including chromosome conformation capture (3C), circular chromosome conformation capture (4C), chromosome conformation capture carbon copy (5C), and high-resolution chromosome conformation capture (Hi-C). The application of these techniques has expanded to various fields, but cancer remains one of the major topics. Interactions mediated by proteins or long noncoding RNAs (lncRNAs) are typically found using 4C-sequencing and chromatin interaction analysis by paired-end tag sequencing (ChIA-PET). Currently, Hi-C is used to identify chromatin loops between cancer risk-associated single-nucleotide polymorphisms (SNPs) found by genome-wide association studies (GWAS) and their target genes. Chromosomal conformations are responsible for altered gene regulation through several typical mechanisms and contribute to the biological behavior and malignancy of different tumors, particularly prostate cancer, breast cancer and hematologic neoplasms. Moreover, different subtypes may exhibit different 3D-chromosomal conformations. Thus, C-tech can be used to help diagnose cancer subtypes and alleviate cancer progression by destroying specific chromosomal conformations. Here, we review the fundamentals and improvements in chromosome conformation capture techniques and their clinical applications in cancer to provide insight for future research.

Sertoli cells are essential for regulating normal spermatogenesis. However, the mechanisms underlying human Sertoli cell development remain largely elusive. Here we examined the function and signaling pathways of BMP6 in regulating human Sertoli cells. RT-PCR, immunocytochemistry and Western blots revealed that BMP6 and its multiple receptors were expressed in human Sertoli cells. CCK-8 and EDU assays showed that BMP6 promoted the proliferation of Sertoli cells. Conversely, BMP6 siRNAs inhibited the division of these cells. Annexin V/PI assay indicated that BMP6 reduced the apoptosis in human Sertoli cells, whereas BMP6 knockdown assumed reverse effects. BMP6 enhanced the expression levels of ZO1, SCF, GDNF and AR in human Sertoli cells, and ELISA assay showed an increase of SCF by BMP6 and a reduction by BMP6 siRNAs. Notably, Smad2/3 phosphorylation and cyclin D1 were enhanced by BMP6 and decreased by BMP6 siRNAs in human Sertoli cells. The levels of DACH1 and TFAP2A were increased by BMP6 and reduced by BMP6 siRNAs, and the growth of human Sertoli cells was inhibited by these siRNAs. Collectively, these results suggest that BMP6 regulates the proliferation and apoptosis of human Sertoli cells via activating the Smad2/3/cyclin D1 and DACH1 and TFAP2A pathway.

Stem cells, by definition, are the primitive cells that have the potential of both self-renewal and differentiation into a number of mature and functional cells, and thus they have great applications in cell therapy and tissue engineering for regenerative medicine. Bone morphogenetic protein 6 (BMP6) belongs to transforming growth factor β(TGF-β) superfamily. The fate determinations of stem cells require complex regulatory networks that involve BMP6 signaling pathway. Recent studies have demonstrated that BMP6 plays crucial roles in controlling the self-renewal and differentiation of stem cells. In this review, we address the expression, function and regulation of BMP6 in various kinds of stem cells, with focus on mesenchymal stem cells (MSCs), germline stem cells (GSCs), hematopoietic stem cells (HSCs), and neural stem cells (NSCs). Notably, there are distinct effects of BMP6 on promoting self-renewal and differentiation of these stem cells. We also discuss the involement of BMP6 in diseases, including leukemia, astrocytic glioma, and Alzheimer's disease, and the therapy of these diseases via gene targeting. We further highlight certain issues for further investigation on the regulation and function of BMP6 in stem cells. Significantly, a thorough understanding of BMP6 regulation on a variety of adult stem cells could make them feasible for applications in both regenerative and reproductive medicine, and it would shed novel insights into the etiology of the diseases and offer new targets for drug design to treat these disorders.