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Obesity and type 2 diabetes are associated with disturbances in insulin-regulated glucose and lipid fluxes and severe comorbidities including cardiovascular disease and steatohepatitis. Whole body metabolism is regulated by lipid-storing white adipocytes as well as “brown” and “brite/beige” adipocytes that express thermogenic uncoupling protein 1 (UCP1) and secrete factors favorable to metabolic health. Implantation of brown fat into obese mice improves glucose tolerance, but translation to humans has been stymied by low abundance of primary human beige adipocytes. Here we apply methods to greatly expand human adipocyte progenitors from small samples of human subcutaneous adipose tissue and then disrupt the thermogenic suppressor gene NRIP1 by CRISPR. Ribonucleoprotein consisting of Cas9 and sgRNA delivered ex vivo are fully degraded by the human cells following high efficiency NRIP1 depletion without detectable off-target editing. Implantation of such CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreases adiposity and liver triglycerides while enhancing glucose tolerance compared to implantation with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic enhancement of human adipocytes without exposing the recipient to immunogenic Cas9 or delivery vectors. Worldwide pandemics of obesity and diabetes prompt an urgent need for new approaches to their prevention and cure. Here the authors present a CRISPR-based strategy that enhances the therapeutic potential of human adipocytes when implanted in obese mice.

Background Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent gastrointestinal malignancies with high mortality. Circular RNAs (CircRNAs) have become a research hotspot in recent years for their vital roles in cancer development and progression. This study aims to clarify the roles of circNTRK2 and its underlying molecular mechanisms in ESCC. Methods The levels of circNTRK2, miR-140-3p, and nuclear receptor-interacting protein 1 (NRIP1) mRNA were examined by qRT-PCR. The cell proliferation ability was detected via CCK-8, EdU and colony formation assays. The invasion capacity was tested by using transwell assay. The apoptotic rate was evaluated through flow cytometry. The protein levels of cleaved PARP, cleaved caspase-3, E-cadherin, vimentin, and NRIP1 were measured by western blot assay. The validation of circular structure was performed by Sanger sequencing, divergent primer PCR, and RNase R treatments. The ceRNA regulatory mechanism of circNTRK2 was observed via dual-luciferase reporter, RIP and RNA pull-down assays. The mice xenograft models were constructed to confirm the oncogenicity of circNTRK2 in ESCC in vivo. Results CircNTRK2 was highly expressed in ESCC tissues and cells. High expression of circNTRK2 was correlated with advanced TNM stage, lymph node metastasis and short survival. Knockdown of circNTRK2 inhibited ESCC cell proliferation, invasion and epithelial-mesenchymal transition (EMT), and accelerated apoptosis in vitro. Mechanistic assays disclosed that circNTRK2 could act as a sponge for miR-140-3p to abate its suppression on target NRIP1 expression. Moreover, miR-140-3p-induced inhibitory effects on ESCC cell malignant phenotypes were attenuated by the overexpression of circNTRK2. In addition, depletion of NRIP1 impeded cell proliferation, invasion and EMT, while enhanced apoptosis. Furthermore, silencing of circNTRK2 suppressed cell proliferation and invasion through regulating NRIP1 expression. Also, knockdown of circNTRK2 slowed ESCC tumor growth in vivo. Conclusion CircNTRK2 promoted ESCC progression by regulating miR-140-3p/NRIP1 pathway. Our findings contribute to a better understanding of circRNAs as miRNA sponges and highlight a promising therapy target in ESCC.

Using chip array assays, we identified differentially expressed genes via a comparison between luminal A breast cancer subtype and normal mammary ductal cells from healthy donors. In silico analysis confirmed by western blot and immunohistochemistry revealed that C-JUN and C-FOS transcription factors are activated in luminal A patients as potential upstream regulators of these differentially expressed genes. Using a chip-on-chip assay, we identified potential C-JUN and C-FOS targets. Among these genes, the NRIP1 gene was revealed to be targeted by C-JUN and C-FOS. This was confirmed after identification and validation with transfection assays specific binding of C-JUN and C-FOS at consensus binding sites. NRIP1 is not only upregulated in luminal A patients and cell lines but also regulates breast cancer-related genes, including PR , ESR1 and CCND1. These results were confirmed by NRIP1 siRNA knockdown and chip array assays, thus highlighting the putative role of NRIP1 in PGR , ESR1 and CCND1 transcriptional regulation and suggesting that NRIP1 could play an important role in breast cancer ductal cell initiation.

During the development of heart failure (HF), the capacity for cardiomyocyte (CM) fatty acid oxidation (FAO) and ATP production is progressively diminished, contributing to pathologic cardiac hypertrophy and contractile dysfunction. Receptor-interacting protein 140 (RIP140, encoded by Nrip1) has been shown to function as a transcriptional corepressor of oxidative metabolism. We found that mice with striated muscle deficiency of RIP140 (strNrip1-/-) exhibited increased expression of a broad array of genes involved in mitochondrial energy metabolism and contractile function in heart and skeletal muscle. strNrip1-/- mice were resistant to the development of pressure overload-induced cardiac hypertrophy, and CM-specific RIP140-deficient (csNrip1-/-) mice were protected against the development of HF caused by pressure overload combined with myocardial infarction. Genomic enhancers activated by RIP140 deficiency in CMs were enriched in binding motifs for transcriptional regulators of mitochondrial function (estrogen-related receptor) and cardiac contractile proteins (myocyte enhancer factor 2). Consistent with a role in the control of cardiac fatty acid oxidation, loss of RIP140 in heart resulted in augmented triacylglyceride turnover and fatty acid utilization. We conclude that RIP140 functions as a suppressor of a transcriptional regulatory network that controls cardiac fuel metabolism and contractile function, representing a potential therapeutic target for the treatment of HF.

Sjögren’s disease (SjD) is marked by dysfunction of the salivary gland (SG) caused by epithelial cell death. However, the mechanism remains unclear. Here we discovered that NRIP1 was abnormally upregulated in SjD and formed a protein complex with estrogen receptor α (ERα) to inhibit saliva secretion and lead to epithelial cell death. NRIP1 interacted with ERα and altered the estradiol (E2)–ERα downstream signal in the SG epithelium. In the context of SjD, NRIP1–ERα suppressed aquaporin-5 (AQP5) expression and promoted MYC expression. The NRIP1–ERα complex bound to the estrogen response elements of the AQP5 promoter, leading to the downregulation of AQP5 expression and reduced SG secretion. Conversely, the NRIP1–ERα complex bound to the estrogen response elements of the MYC promoter, resulting in the upregulation of MYC expression. Furthermore, we demonstrated that elevated MYC levels promoted apoptosis and altered immune regulation and cell metabolism in SjD. Nrip1 -knockout/ovariectomized mice did not develop the SjD phenotypes, confirming the role of NRIP1 in the pathophysiology of SjD. Molecular dynamic simulations revealed that NRIP1 competitively bound to ERα and masked the E2 binding site, providing structural insights into the disruption of hormonal signal. This study implicates NRIP1 as a potent diagnosis parameter and provides a putative target for SjD management. NRIP1 complex impairs saliva secretion in Sjögren’s disease This study explores Sjögren’s syndrome (SjD), a condition causing dry mouth and eyes, often affecting postmenopausal women. Researchers aimed to understand why salivary gland cells malfunction in SjD. The team found that a protein called NRIP1 is overactive in these cells during SjD. They used mouse models and cell experiments to study this. NRIP1 interacts with another protein, ERα, affecting two key genes: AQP5, which helps with saliva production, and MYC, which influences cell survival and immune response. In SjD, NRIP1 reduces AQP5, leading to less saliva, and increases MYC, causing cell damage. The researchers also discovered that removing NRIP1 in mice prevented SjD symptoms. This suggests that targeting NRIP1 could be a new way to treat SjD. Future research may focus on developing therapies that adjust NRIP1 activity to improve symptoms in patients with SjD. This summary was initially drafted using artificial intelligence, then revised and fact-checked by the author. SjD-related phlogosis not only switches off the E2–ERα signal but also activates the NRIP1–ERα signal that downregulates AQP5 expression and upregulates MYC expression to aggravate SjD.

Glioma is one of the most common and aggressive types of brain tumors, characterized by generally low survival rates. Circular RNAs (circRNAs) have been identified as key players in the development of glioma. Our study aims to investigate the effect of circNRIP1 on glioma progression and elucidate the underlying mechanisms involved. The mRNA expression of circNRIP1, miR-106a-5p, and GPR133 was determined using qRT-PCR. The protein expression of N-cadherin, E-cadherin, fibronectin, vimentin, and GPR133 in glioma cells and tissues was measured through Western blotting and IHC assays. Cell counting kit-8 (CCK-8), wound healing, and transwell assays were used to determine cell proliferation, migration, and invasion, respectively. The dual-luciferase reporter assay was utilized to elucidate the targeting interaction between miR-106a-5p and both circNRIP1 and GPR133. Finally, xenograft models were established to investigate the effects of circNRIP1 in vivo. The expression of circNRIP1 was significantly elevated in glioma cell lines and tissues, and this higher expression correlated with a reduced overall survival rate. Downregulation of circNRIP1 had been shown to inhibit the invasion, migration, proliferation, and epithelial-mesenchymal transition (EMT) of glioma cells and attenuate tumor growth in vivo. circNRIP1 functions as a molecular sponge for miR-106a-5p, consequently elevating the expression of GPR133. Additionally, upregulation of GPR133 could counteract the inhibition of the malignant phenotype of glioma cells caused by circNRIP1 knockdown. circNRIP1 knockdown suppresses glioma progression by elevating miR-106a-5p levels and simultaneously reducing GPR133 expression. The identified circNRIP1/miR-106a-5p/GPR133 axis could potentially be a therapeutic target for glioma.

Esophageal squamous cell carcinoma (ESCC) is characterized by a complex tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) play a crucial role in the TME that facilitate tumor progression via interactions with cancer cells. However, the mechanisms underlying the activation of CAFs in TME remain largely unknown. Here, we characterized the exosomes derived from normoxic and hypoxic ESCC cells using electron microscopy and western blot. The impact of exosomes on CAF activation and the motility of ESCC cells was examined in vitro. The molecular complex involving circNRIP1 was explored using RNA pull-down. We demonstrated that exosomes derived from ESCC cells, including KYSE-150 and TE-10 cells, exhibited a significantly increase in secretion under hypoxic conditions. These hypoxic exosomes were internalized by fibroblasts and further promoted the transformation of normal fibroblasts into CAFs, as evidenced by enhanced migration and secretion of pro-inflammatory cytokines. circNRIP1 was enriched in hypoxic exosomes, and its absence abolished the effect of hypoxic exosomes to activate CAFs. Furthermore, the CAFs activated by exosomal circNRIP1 further promoted the migration and invasion of ESCC cells. Mechanistically, circNRIP1 bound to the N1-methyladenosine (m1A) methyltransferase TRMT6 and activated CAFs in a TRMT6-dependent manner. This study revealed the role of hypoxia-induced exosomal circNRIP1 in the activation of CAFs, which contributes to ESCC development. These findings shed light on the mechanisms of the CAF activation in ESCC, positioning hypoxia-induced exosomal circNRIP1 as a potential molecular target for ESCC.

Background Genome-wide association studies (GWASs) have identified numerous single-nucleotide polymorphisms (SNPs) associated with small increases in breast cancer risk. Studies to date suggest that some SNPs alter the expression of the associated genes, which potentially mediates risk modification. On this basis, we hypothesised that some of these genes may be enriched for rare coding variants associated with a higher breast cancer risk. Methods The coding regions and exon-intron boundaries of 56 genes that have either been proposed by GWASs to be the regulatory targets of the SNPs and/or located < 500 kb from the risk SNPs were sequenced in index cases from 1043 familial breast cancer families that previously had negative test results for BRCA1 and BRCA2 mutations and 944 population-matched cancer-free control participants from an Australian population. Rare (minor allele frequency ≤ 0.001 in the Exome Aggregation Consortium and Exome Variant Server databases) loss-of-function (LoF) and missense variants were studied. Results LoF variants were rare in both the cases and control participants across all the candidate genes, with only 38 different LoF variants observed in a total of 39 carriers. For the majority of genes ( n  = 36), no LoF variants were detected in either the case or control cohorts. No individual gene showed a significant excess of LoF or missense variants in the cases compared with control participants. Among all candidate genes as a group, the total number of carriers with LoF variants was higher in the cases than in the control participants (26 cases and 13 control participants), as was the total number of carriers with missense variants (406 versus 353), but neither reached statistical significance ( p  = 0.077 and p  = 0.512, respectively). The genes contributing most of the excess of LoF variants in the cases included TET2 , NRIP1 , RAD51B and SNX32 (12 cases versus 2 control participants), whereas ZNF283 and CASP8 contributed largely to the excess of missense variants (25 cases versus 8 control participants). Conclusions Our data suggest that rare LoF and missense variants in genes associated with low-penetrance breast cancer risk SNPs may contribute some additional risk, but as a group these genes are unlikely to be major contributors to breast cancer heritability.

PurposeLigand-dependent corepressor (LCoR) and receptor-interacting protein 140 (RIP140/NRIP1) play an important role in the regulation of multiple oncogenic signaling pathways and the development of cancer. LCoR and RIP140 form a nuclear complex in breast cancer cells and are of prognostic value in further prostate and cervical cancer. The purpose of this study was to analyze the regulation of these proteins in the development of cervical intraepithelial neoplasia (CIN I–III).MethodsImmunohistochemical analysis was obtained to quantify RIP140 and LCoR expression in formalin-fixed paraffin embedded tissue sections of cervical intraepithelial neoplasia samples. Tissue (n = 94) was collected from patients treated in the Department of Gynecology and Obstetrics, Ludwig-Maximilians-University of Munich, Germany, between 2002 and 2014. Correlations of expression levels with clinical outcome were carried out to assess for prognostic relevance in patients with CIN2 progression. Kruskal–Wallis test and Mann–Whitney U test were used for data analysis.ResultsNuclear LCoR overexpression correlates significantly with CIN II progression. Nuclear RIP140 expression significantly increases and nuclear LCoR expression decreases with higher grading of cervical intraepithelial neoplasia. Cytoplasmic RIP140 expression is significantly higher in CIN III than in CIN I or CIN II.ConclusionA decrease of nuclear LCoR expression in line with an increase of dedifferentiation of CIN can be observed. Nuclear LCoR overexpression correlates with CIN II progression indicating a prognostic value of LCoR in cervical intraepithelial neoplasia. Nuclear and cytoplasmic RIP140 expression increases significantly with higher grading of cervical intraepithelial neoplasia underlining its potential role in the development of pre-cancerous lesions. These findings support the relevance of LCoR and RIP140 in the tumorigenesis indicating a possible role of LCoR and RIP140 as targets for novel therapeutic approaches in cervical intraepithelial neoplasia and cervical cancer.

The present study aimed to investigate the influence of circular RNA nuclear receptor-interacting protein 1 (circNRIP1) on the chemotherapeutic effect of 5-fluorouracil (5-FU) in colorectal cancer (CRC) and reveal its potential molecular mechanisms. The effects of circNRIP1 on cell proliferation, migration and invasion, and apoptosis were evaluated using Cell Counting Kit-8, Transwell and flow cytometric assays, respectively. A dual-luciferase reporter assay was performed to verify the potential interaction between circNRIP1 and microRNA (miR)-532-3p. The results of the present study indicated that circNRIP1 was upregulated in CRC and its increased expression was associated with CRC progression. Furthermore, overexpression of circNRIP1 promoted CRC cell proliferation, invasion and migration, while it inhibited apoptosis. Knockdown of circNRIP1 significantly enhanced the 5-FU-induced inhibition of the viability of HCT116 and SW480 cells. Bioinformatics analysis predicted that miR-532-3p was a direct target of circNRIP1, which was further confirmed by a dual-luciferase reporter assay. miR-532-3p silencing reversed the effects of circNRIP1 knockdown on the sensitivity of 5-FU in the chemotherapy of CRC. The results suggested that circNRIP1 and miR-532-3p may be utilized to improve the diagnosis of CRC and serve as diagnostic markers. In conclusion, overexpression of circNRIP1 promoted the progression of CRC, while circNRIP1 silencing sensitized CRC cells to 5-FU via sponging miR-532-3p.