FTO Deficiency Aggravates Ulcerative Colitis via Disrupting CerS6-Mediated Sphingolipid Metabolism: The Empowering Role of ANT BIO PTE. LTD.’s Absin Multiplex IHC Kits

Ulcerative Colitis (UC), a major subtype of inflammatory bowel disease (IBD), is a chronic, relapsing inflammatory disorder characterized by persistent inflammation and ulceration of the colonic mucosa. The pathogenesis of UC involves complex interactions between genetic factors, environmental triggers, and immune dysregulation. N6-methyladenosine (m6A) modification, the most prevalent post-transcriptional modification of eukaryotic mRNA, has been implicated in various inflammatory diseases. Fat mass and obesity-associated protein (FTO), a nuclear-localized m6A demethylase, exerts its biological function by reducing m6A levels through oxidative demethylase activity, specifically targeting m6A in single-stranded RNA. Emerging evidence suggests that m6A modification participates in the pathogenesis of UC, but the specific role and underlying mechanism of FTO in UC remain unclear.

On September 21, 2023, a joint research team from Jie Hong and Haoyan Chen of Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, and Ruixin Liu of Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, published a research paper entitled "Disruption of CerS6-Mediated Sphingolipid Metabolism by FTO Deficiency Aggravates Ulcerative Colitis" in Gut. The study revealed that downregulation of FTO promotes the occurrence of UC by reducing the expression of Ceramide Synthase 6 (CerS6), leading to the accumulation of Sphingosine-1-Phosphate (S1P) in intestinal epithelial cells (IECs), and exacerbates UC through an m6A-dependent mechanism. Additionally, the research team found that reduced FTO expression in UC patients may enhance their response to the monoclonal antibody drug vedolizumab. Notably, multiplex immunohistochemistry (mIHC) kits from the Absin product line of ANT BIO PTE. LTD. played a crucial role in validating the expression of integrin α4β7 (the target of vedolizumab) in Th17 cells of mouse models, providing direct experimental evidence for the correlation between FTO expression and vedolizumab therapeutic efficacy.

1. Literature Information

•         Title: Disruption of CerS6-Mediated Sphingolipid Metabolism by FTO Deficiency Aggravates Ulcerative Colitis

•         Journal: Gut

•         Publication Date: Online on September 21, 2023 (Epub ahead of print)

•         DOI: 10.1136/gutjnl-2023-330009

•         Research Team: Jie Hong, Haoyan Chen et al., Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine; Ruixin Liu et al., Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

•         PMID: 37734910

•         Core Reagents from ANT BIO PTE. LTD.: Multiplex Fluorescence IHC Staining Kits (Absin product line), including 4-color to 7-color plus kits (e.g., Catalog No.: abs50012, abs50015) and supporting reagent Antibody Elution Buffer (mIHC-Specific, Catalog No.: abs994)

•         Related Product Link: AntBio - Fueling Research, Feeding Discovery

2. Research Background

UC is a chronic inflammatory bowel disease with increasing incidence worldwide, severely affecting patients' quality of life. The complex pathogenesis of UC involves immune dysfunction, intestinal epithelial barrier damage, and abnormal metabolic regulation. m6A modification, as a key post-transcriptional regulatory mechanism, participates in the regulation of gene expression, RNA stability, and translation efficiency, and is closely associated with the occurrence and development of inflammatory diseases. FTO, as an m6A demethylase, has been reported to be involved in the regulation of various physiological and pathological processes, but its role in UC and the underlying molecular mechanisms have not been fully elucidated. In addition, the clinical efficacy of biological agents for UC varies among patients, and there is an urgent need to identify reliable biomarkers to predict therapeutic responses and guide personalized treatment.

3. Research Strategy

The research team adopted a multi-dimensional approach combining clinical sample analysis, animal models, and molecular biology experiments to systematically explore the role and mechanism of FTO in UC, as well as its correlation with therapeutic efficacy:

1.       Clinical Sample Analysis: Collected intestinal mucosal tissues from UC patients and non-inflammatory control groups. Performed RNA sequencing analysis to detect the expression level of FTO in intestinal tissues. Conducted functional analysis, Western Blot (WB), and immunofluorescence experiments to verify the role of FTO as an m6A-related gene in the occurrence and development of UC.

2.       Animal Model Construction and Phenotype Detection: Constructed intestinal epithelium-specific FTO knockout mice (Fto; Villin-cre). Induced colitis models using Dextran Sodium Sulfate (DSS). Compared the severity of colitis phenotypes between FTO conditional knockout mice and control mice.

3.       Molecular Mechanism Exploration: Identified the main target genes of FTO in intestinal epithelial cells through molecular biology experiments. Explored the regulatory effect of FTO on CerS6-mediated sphingolipid metabolism and its impact on S1P accumulation in intestinal epithelial cells. Investigated the role of FTO in regulating pro-inflammatory macrophages through endothelial cell-secreted S1P.

4.       Correlation Analysis Between FTO Expression and Therapeutic Efficacy: Searched for data on biological agent treatment of UC patients in the GEO database to analyze the correlation between FTO expression level and therapeutic response to vedolizumab. Performed IHC experiments on clinical samples to compare the effective rate of vedolizumab treatment between the low-FTO expression group and the high-FTO expression group.

5.       mIHC Validation with Absin Kits: Used ANT BIO PTE. LTD.’s Absin multiplex IHC kits to detect the expression of integrin α4β7 (the target of vedolizumab) in Th17 cells of Fto-DSS mice and Fto; Villin-Cre-DSS mice, verifying the correlation between FTO expression and the expression of vedolizumab target molecules.

4. Key Research Findings

4.1 FTO Expression is Persistently Reduced in Intestinal Tissues of UC Patients

RNA sequencing analysis of intestinal mucosal tissues from UC patients and non-inflammatory control groups showed that the level of FTO in the intestinal tissues of UC patients was persistently reduced. Functional analysis, WB, and immunofluorescence experiments further confirmed that FTO, as an m6A-related gene, may play an important role in the occurrence and development of UC. This finding suggests that FTO downregulation may be closely associated with the pathogenesis of UC.

4.2 Intestinal Epithelium-Specific FTO Knockout Aggravates DSS-Induced Colitis in Mice

The research team successfully constructed intestinal epithelium-specific FTO knockout mice (Fto; Villin-cre) and induced colitis models using DSS. The results showed that compared with control mice (Fto), FTO conditional knockout mice exhibited more severe colitis phenotypes, including more obvious weight loss, intestinal mucosal damage, and inflammatory cell infiltration. This in vivo experiment confirms that FTO deficiency exacerbates UC progression.

4.3 FTO Deficiency Disrupts CerS6-Mediated Sphingolipid Metabolism to Aggravate UC via m6A-Dependent Mechanism

Mechanistic studies identified CerS6 as the main target gene of FTO in intestinal epithelial cells. FTO downregulation reduces the expression of CerS6, leading to the accumulation of S1P in intestinal epithelial cells. Further experiments confirmed that FTO regulates CerS6 expression through an m6A-dependent mechanism, and the accumulated S1P regulates pro-inflammatory macrophages, thereby exacerbating intestinal inflammation. This finding reveals a novel molecular pathway by which FTO deficiency contributes to UC progression.

4.4 Low FTO Expression Enhances UC Patients' Response to Vedolizumab Treatment

To explore more suitable treatment options for UC patients, the research team analyzed data on biological agent treatment of UC patients in the GEO database. The results showed that UC patients with low FTO expression were more likely to respond to vedolizumab treatment, but not to other biological agents. IHC results of clinical samples further confirmed that the effective rate of vedolizumab treatment in the low-FTO expression group was significantly higher than that in the high-FTO expression group.

To verify the underlying mechanism, the research team used ANT BIO PTE. LTD.’s Absin multiplex IHC kits to detect the expression of integrin α4β7 (the target of vedolizumab) in Th17 cells of mouse models. The results showed that the expression of integrin α4β7 in Th17 cells of Fto; Villin-Cre-DSS mice was significantly higher than that of Fto-DSS mice. These results collectively indicate that UC patients with low FTO expression have better therapeutic effects on vedolizumab.

5. Product Empowerment: The Critical Role of ANT BIO PTE. LTD.’s Absin Multiplex IHC Kits

Validating the expression difference of integrin α4β7 (vedolizumab target) in Th17 cells between FTO knockout and control mice was a key step in this study, as it provided direct experimental evidence for the correlation between FTO expression and vedolizumab therapeutic efficacy. ANT BIO PTE. LTD.’s Absin multiplex fluorescence IHC kits delivered the high-performance multiplexing capability, sensitivity, and specificity required for this critical validation, enabling the simultaneous detection of multiple molecular markers in mouse intestinal tissue sections while preserving the spatial context of their expression.

5.1 Core Products and Their Application Value

5.2 Technical Value in Inflammatory Bowel Disease Research

In inflammatory bowel disease research, the accurate detection of the spatial distribution and expression level of key molecules in specific cell subsets is crucial for deciphering the pathogenesis and exploring therapeutic targets. ANT BIO PTE. LTD.’s Absin multiplex fluorescence IHC kits possess unique advantages in this field compared with traditional single-marker IHC. Their ability to simultaneously detect multiple key molecules on a single tissue section preserves the native spatial context of cell-cell interactions and molecular expression, which is essential for exploring the complex regulatory networks in inflammatory bowel diseases.

In this UC research, the high sensitivity and specificity of the Absin kits ensured clear detection of low-abundance molecules (such as integrin α4β7) in complex intestinal tissue samples. The kits’ compatibility with animal model tissues and clinical samples also supports the translational application of research findings from the laboratory to the clinic. This technical empowerment was instrumental in validating the correlation between FTO expression and vedolizumab therapeutic efficacy, highlighting the important value of ANT BIO PTE. LTD.’s products in promoting inflammatory bowel disease research and guiding personalized treatment.

6. Brand Mission

As a professional supplier of life science reagents, ANT BIO PTE. LTD. is dedicated to providing high-quality, reliable products and comprehensive solutions to empower global life science research. The company's three specialized sub-brands cover the full spectrum of research needs in the life science field: Absin focuses on general reagents and kits, Starter specializes in antibodies, and UA is dedicated to recombinant proteins. Our core mission is to bridge the gap between cutting-edge scientific research and practical applications, accelerate the pace of scientific discovery, and contribute to the advancement of human health and regenerative medicine.

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8. Disclaimer

This article is AI-compiled and interpreted based on the original work in DOI: 10.1136/gutjnl-2023-330009. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

9. Brand Promotion Copy

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At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. The Absin multiplex fluorescence IHC kits, with their unique advantages of breaking species limitations and supporting multi-marker simultaneous detection, provide powerful technical support for inflammatory bowel disease research, immunology, and precision medicine. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.