Bach2 Deficiency Promotes Intestinal Epithelial Regeneration by Accelerating DNA Repair in Intestinal Stem Cells

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gut. Available drugs aim to suppress gut inflammation. These drugs have significantly delayed disease progression and improved patients’ quality of life. However, the disease continues to progress, underscoring the need to develop novel therapies. Aside from chronic gut inflammation, IBD patients also experience a leaky gut problem due to damage to the intestinal epithelial layer. In this regard, epithelial regeneration and repair are mediated by intestinal stem cells. However, no therapies are available to directly enhance the intestinal stem cells’ regenerative and repair function. Recently, it was shown that active vitamin D, i.e., 1,25-dihydroxyvitamin D or 1,25(OH)2D, was necessary to maintain Lgr5+ intestinal stem cells, actively cycling under physiological conditions. In this study, we used two strategies to investigate the role of 1,25(OH)2D in intestinal stem cells’ regenerative function. First, to avoid the side effects of systemic high 1,25(OH)2D conditions, we used our recently developed novel strategy to deliver locally high 1,25(OH)2D concentrations specifically to inflamed intestines. Second, because of the Lgr5+ intestinal stem cells’ active cycling status, we used a pulse-and-chase strategy via 5-bromo-2′-deoxyuridine (BrdU) labeling to trace the Lgr5+ stem cells through the whole epithelial regeneration process. Our data showed that locally high 1,25(OH)2D concentrations enhanced intestinal stem cell migration. Additionally, the migrated cells differentiated into mature epithelial cells. Our data, therefore, suggest that local delivery of high 1,25(OH)2D concentrations is a promising strategy to augment intestinal epithelial repair in IBD patients.

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Epithelial Regeneration after Doxorubicin Arises Primarily from Early Progeny of Active Intestinal Stem Cells

SSRN Electronic Journal ◽

10.2139/ssrn.3593526 ◽

2020 ◽

Author(s):

Breanna Sheahan ◽

Ally N. Freeman ◽

Theresa M. Keeley ◽

Linda C. Samuelson ◽

Jatin Roper ◽

...

Keyword(s):

Stem Cells ◽

Intestinal Stem Cells ◽

Epithelial Regeneration

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Unveiling role of Sphingosine-1-phosphate receptor 2 as a brake of epithelial stem cell proliferation and a tumor suppressor in colorectal cancer

10.21203/rs.3.rs-56319/v1 ◽

2020 ◽

Author(s):

Luciana Petti ◽

Giulia Rizzo ◽

Federica Rubbino ◽

Sudharshan Elangovan ◽

Piergiuseppe Colombo ◽

...

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Normal Mucosa ◽

Genetically Engineered ◽

Colorectal Carcinogenesis ◽

Intestinal Stem Cells ◽

Cancer Information ◽

Intestinal Epithelial ◽

Epithelial Stem Cells ◽

Mucosal Regeneration

Abstract BackgroundSphingosine-1-phosphate receptor 2 (S1PR2) mediates pleiotropic functions encompassing cell proliferation, survival, and migration, which become collectively de-regulated in cancer. Information onto whether S1PR2 participates in colorectal carcinogenesis/cancer is scanty, and we set out to fill the gap.MethodsWe screened expression changes of S1PR2 in human CRC and matched normal mucosa specimens [N = 76]. We compared CRC arising in inflammation-driven and genetically engineered models in wild-type (S1PR2+/+) and S1PR2 deficient (S1PR2−/−) mice. We reconstituted S1PR2 expression in RKO cells and assessed their growth in xenografts. Functionally, we mimicked ablation of S1PR2 in normal mucosa by treating S1PR2+/+ organoids with JTE013, and characterized intestinal epithelial stem cells isolated from S1PR2−/−Lgr5-EGFP- mice.ResultsS1PR2 expression was lost in 33% of CRC; in 55%, it was significantly decreased, only 12% retaining expression comparable to normal mucosa. Both colitis-induced and genetic Apc+/min mouse models of CRC showed a higher incidence in size and number of carcinomas and/or high-grade adenomas, with increased cell proliferation in S1PR2−/− mice compared to S1PR2+/+ controls. Loss of S1PR2 impaired mucosal regeneration, ultimately promoting the expansion of intestinal stem cells. Whereas its overexpression attenuated cell cycle progression, it reduced the phosphorylation of AKT and augmented the levels of PTEN.ConclusionsIn normal colonic crypts, S1PR2 gains expression along with intestinal epithelial cells differentiation, but not in intestinal stem cells, and contrasts intestinal tumorigenesis by promoting epithelial differentiation, preventing the expansion of stem cells and braking their malignant transformation. Targeting of S1PR2 may be of therapeutic benefit for CRC expressing high Lgr5.

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Unveiling role of sphingosine-1-phosphate receptor 2 as a brake of epithelial stem cell proliferation and a tumor suppressor in colorectal cancer

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-020-01740-6 ◽

2020 ◽

Vol 39 (1) ◽

Author(s):

Luciana Petti ◽

Giulia Rizzo ◽

Federica Rubbino ◽

Sudharshan Elangovan ◽

Piergiuseppe Colombo ◽

...

Keyword(s):

Colorectal Cancer ◽

Stem Cells ◽

Cell Proliferation ◽

Stem Cell ◽

Tumor Suppressor ◽

Normal Mucosa ◽

Intestinal Stem Cells ◽

Intestinal Epithelial ◽

Sphingosine 1 Phosphate

Abstract Background Sphingosine-1-phosphate receptor 2 (S1PR2) mediates pleiotropic functions encompassing cell proliferation, survival, and migration, which become collectively de-regulated in cancer. Information on whether S1PR2 participates in colorectal carcinogenesis/cancer is scanty, and we set out to fill the gap. Methods We screened expression changes of S1PR2 in human CRC and matched normal mucosa specimens [N = 76]. We compared CRC arising in inflammation-driven and genetically engineered models in wild-type (S1PR2+/+) and S1PR2 deficient (S1PR2−/−) mice. We reconstituted S1PR2 expression in RKO cells and assessed their growth in xenografts. Functionally, we mimicked the ablation of S1PR2 in normal mucosa by treating S1PR2+/+ organoids with JTE013 and characterized intestinal epithelial stem cells isolated from S1PR2−/−Lgr5-EGFP- mice. Results S1PR2 expression was lost in 33% of CRC; in 55%, it was significantly decreased, only 12% retaining expression comparable to normal mucosa. Both colitis-induced and genetic Apc+/min mouse models of CRC showed a higher incidence in size and number of carcinomas and/or high-grade adenomas, with increased cell proliferation in S1PR2−/− mice compared to S1PR2+/+ controls. Loss of S1PR2 impaired mucosal regeneration, ultimately promoting the expansion of intestinal stem cells. Whereas its overexpression attenuated cell cycle progression, it reduced the phosphorylation of AKT and augmented the levels of PTEN. Conclusions In normal colonic crypts, S1PR2 gains expression along with intestinal epithelial cells differentiation, but not in intestinal stem cells, and contrasts intestinal tumorigenesis by promoting epithelial differentiation, preventing the expansion of stem cells and braking their malignant transformation. Targeting of S1PR2 may be of therapeutic benefit for CRC expressing high Lgr5. Graphical Abstract. Schematic drawing of the role of S1PR2 in normal mucosa and colorectal cancer. In the normal mucosa, S1PR2 is highly expressed by differentiated cells at the upper region of both colon and intestinal crypts (S1PR2 ON), but not by the undifferentiated stem cell at the base of the crypts (S1PR2 OFF), in which acts as a negative proliferative regulator promoting epithelial differentiation. Its loss leads to the expansion of stem cells and reduced levels of PTEN and Axin-2, two negative regulators respectively of PI3K/AKT and Wnt signaling that control β-catenin signaling. The translocation of β-catenin into the nucleus promotes the transcription of target genes involved in the proliferation and malignant transformation. Thereby, S1PR2 works in the intestine as a tumor suppressor

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Intestinal epithelial cell-specific IGF1 promotes the expansion of intestinal stem cells during epithelial regeneration and functions on the intestinal immune homeostasis

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00022.2018 ◽

2018 ◽

Vol 315 (4) ◽

pp. E638-E649 ◽

Cited By ~ 5

Author(s):

Yu Zheng ◽

Yongli Song ◽

Qi Han ◽

Wenjie Liu ◽

Jiuzhi Xu ◽

...

Keyword(s):

Stem Cells ◽

Epithelial Cell ◽

Intestinal Epithelial Cell ◽

High Throughput Sequencing ◽

Secretory Cells ◽

Intestinal Epithelial ◽

Epithelial Stem Cells ◽

Microbial Composition ◽

Epithelial Regeneration

It is well known that insulin-like growth factor 1 (IGF1) acts as a trophic factor in small intestine under both physiological and pathophysiological conditions. However, it still lacks direct in vivo evidence of the functions of intestinal epithelial cell (IEC)-specific IGF1 under both normal and pathological conditions. Using IEC-specific IGF1-knockout (cKO) mice and Lgr5-eGFP-CreERT mice, we demonstrate that IEC-specific IGF1 can enhance nutrient uptake, reduce protein catabolism and energy consumption, and promote the proliferation and expansion of intestinal epithelial cells, including intestinal epithelial stem cells and intestinal secretory cells. Next, we showed that IEC-specific IGF1 renders IECs resistant to irradiation and promotes epithelial regeneration. Strikingly, transcriptome profiling assay revealed that many differentially expressed genes involved in the differentiation and maturation of lymphoid lineages were significantly suppressed in the cKO mice as compared with the control mice. We demonstrated that deletion of IGF1 in IECs enhances bacterial translocation to the mesenteric lymph nodes and liver. Furthermore, high-throughput sequencing of 16S ribosomal RNA genes of gut microbiota revealed that IEC-specific IGF1 loss profoundly affected the gut microbial composition at various levels of classification. Therefore, our findings shed light on the in vivo roles of IEC-specific IGF1 in intestinal homeostasis, epithelial regeneration, and immunity, broadening our current insights on IGF1 functions.

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Intestinal epithelial regeneration: active versus reserve stem cells and plasticity mechanisms

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00126.2019 ◽

2020 ◽

Vol 318 (4) ◽

pp. G796-G802 ◽

Cited By ~ 2

Author(s):

Soham Karmakar ◽

Lu Deng ◽

Xi C. He ◽

Linheng Li

Keyword(s):

Stem Cells ◽

Small Intestine ◽

Large Intestine ◽

Definitive Endoderm ◽

Intestinal Epithelial ◽

Developmental Systems ◽

Epithelial Regeneration ◽

Colon And Rectum ◽

Food Absorption ◽

Further Development

The gastrointestinal system is arguably one of the most complicated developmental systems in a multicellular organism, as it carries out at least four major functions: digestion of food, absorption of nutrients, excretion of hormones, and defense against pathogens. Anatomically, the fetal gut has a tubular structure with an outer layer of smooth muscle derived from lateral splanchnic mesoderm and an inner lining of epithelium derived from the definitive endoderm. During morphogenesis of the gut tube, the definitive endoderm transforms into a primitive gut tube with a foregut, midgut, and hindgut. During the course of further development, the midgut gives rise to the small and proximal large intestine and the hindgut gives rise to the distal large intestine and rectum. The small intestine is subdivided into three parts: duodenum, jejunum, and ileum, whereas the large intestine is subdivided into the cecum, colon, and rectum.

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Source and Impact of the EGF Family of Ligands on Intestinal Stem Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.685665 ◽

2021 ◽

Vol 9 ◽

Author(s):

Helen E. Abud ◽

Wing Hei Chan ◽

Thierry Jardé

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Ex Vivo ◽

Intestinal Stem Cells ◽

Intestinal Epithelial ◽

Neuregulin 1 ◽

Epithelial Development ◽

Epidermal Growth ◽

Egf Family

Epidermal Growth Factor (EGF) has long been known for its role in promoting proliferation of intestinal epithelial cells. EGF is produced by epithelial niche cells at the base of crypts in vivo and is routinely added to the culture medium to support the growth of intestinal organoids ex vivo. The recent identification of diverse stromal cell populations that reside underneath intestinal crypts has enabled the characterization of key growth factor cues supplied by these cells. The nature of these signals and how they are delivered to drive intestinal epithelial development, daily homeostasis and tissue regeneration following injury are being investigated. It is clear that aside from EGF, other ligands of the family, including Neuregulin 1 (NRG1), have distinct roles in supporting the function of intestinal stem cells through the ErbB pathway.

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Qingchang Wenzhong Decoction Accelerates Intestinal Mucosal Healing Through Modulation of Dysregulated Gut Microbiome, Intestinal Barrier and Immune Responses in Mice

Frontiers in Pharmacology ◽

10.3389/fphar.2021.738152 ◽

2021 ◽

Vol 12 ◽

Author(s):

Zhongmei Sun ◽

Junxiang Li ◽

Wenting Wang ◽

Yuyue Liu ◽

Jia Liu ◽

...

Keyword(s):

Stem Cells ◽

Gut Microbiota ◽

Intestinal Inflammation ◽

Intestinal Barrier ◽

Mucosal Healing ◽

Intestinal Stem Cells ◽

Mucosal Inflammation ◽

Protective Effects ◽

Fecal Microbiome ◽

Epithelial Regeneration

Inflammatory bowel disease (IBD), a group of multifactorial and inflammatory infirmities, is closely associated with dysregulation of gut microbiota and host metabolome, but effective treatments are currently limited. Qingchang Wenzhong Decoction (QCWZD) is an effective and classical traditional herbal prescription for the treatment of IBD and has been proved to attenuate intestinal inflammation in a model of acute colitis. However, the role of QCWZD in recovery phase of colitis is unclear. Here, we demonstrated that mice treated with QCWZD showed a faster recovery from dextran sulfate sodium (DSS)-induced epithelial injury, accompanied by reduced mucosal inflammation and attenuated intestinal dysbiosis using bacterial 16S rRNA amplicon sequencing compared to those receiving sterile water. The protective effects of QCWZD are gut microbiota dependent, as demonstrated by fecal microbiome transplantation and antibiotics treatment. Gut microbes transferred from QCWZD-treated mice displayed a similar role in mucosal protection and epithelial regeneration as QCWZD on colitis in mice, and depletion of the gut microbiota through antibiotics treatments diminished the beneficial effects of QCWZD on colitis mice. Moreover, metabolomic analysis revealed metabolic profiles alternations in response to the gut microbiota reprogrammed by QCWZD intervention, especially enhanced tryptophan metabolism, which may further accelerate intestinal stem cells-mediated epithelial regeneration to protect the integrity of intestinal mucosa through activation of Wnt/β-catenin signals. Collectively, our results suggested that orally administrated QCWZD accelerates intestinal mucosal healing through the modulation of dysregulated gut microbiota and metabolism, thus regulating intestinal stem cells-mediated epithelial proliferation, and hold promise for novel microbial-based therapies in the treatment of IBD.

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PTBP1/HNRNP I controls intestinal epithelial cell regeneration by maintaining stem cell survival and stemness

10.1101/2021.11.03.466345 ◽

2021 ◽

Author(s):

Wesley Tung ◽

Ullas Valiya Chembazhi ◽

Jing Yang ◽

Ka Lam Nguyen ◽

Aryan Lalwani ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Epithelial Cell ◽

Cell Survival ◽

Intestinal Epithelial Cell ◽

Intestinal Stem Cells ◽

Intestinal Stem Cell ◽

Cell Regeneration ◽

Intestinal Epithelial ◽

Stem Cell Survival

Properly controlled intestinal epithelial cell regeneration is not only vital for protection against insults from environmental hazards but also crucial for preventing intestinal cancer. Intestinal stem cells located in the crypt region provide the driving force for epithelial regeneration, and thus their survival and death must be precisely regulated. We show here that polypyrimidine tract binding protein 1 (PTBP1, also called heterogeneous nuclear ribonucleoprotein I, or HNRNP I), an RNA-binding protein that post-transcriptionally regulates gene expression, is critical for intestinal stem cell survival and stemness. Mechanistically, we show that PTBP1 inhibits the expression of PHLDA3, an AKT repressor, and thereby maintains AKT activity in the intestinal stem cell compartment to promote stem cell survival and proliferation. Furthermore, we show that PTBP1 inhibits the expression of PTBP2, a paralog of PTBP1 that is known to induce neuron differentiation, through repressing inclusion of alternative exon 10 to Ptbp2 transcript. Loss of PTBP1 results in a significant upregulation of PTBP2, which is accompanied by splicing changes in genes that are important for neuron cell development. This finding suggests that PTBP1 prevents aberrant differentiation of intestinal stem cells into neuronal cells through inhibiting PTBP2. Our results thus reveal a novel mechanism whereby PTBP1 maintains intestinal stem cell survival and stemness through the control of gene function post-transcriptionally.

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Lactobacillus plantarum IS-10506 activates intestinal stem cells in a rodent model

Beneficial Microbes ◽

10.3920/bm2017.0118 ◽

2018 ◽

Vol 9 (5) ◽

pp. 755-760 ◽

Cited By ~ 2

Author(s):

A.F. Athiyyah ◽

A. Darma ◽

R. Ranuh ◽

W. Riawan ◽

A. Endaryanto ◽

...

Keyword(s):

Stem Cells ◽

Lactobacillus Plantarum ◽

Gastric Tube ◽

Intestinal Stem Cells ◽

Intestinal Epithelial ◽

Sprague Dawley ◽

Leukaemia Virus ◽

Extracellular Signal ◽

Probiotic Treatment ◽

Lgr5 Expression

This study investigated the probiotic effect of Lactobacillus plantarum IS-10506 in activating and regenerating leucine-rich repeat-containing G-protein-coupled receptor (Lgr)5- and B lymphoma Moloney murine leukaemia virus insertion region (Bmi)1-expressing intestinal stem cells in rodents following Escherichia coli serotype 055:B5 lipopolysaccharide (LPS) exposure. Male Sprague-Dawley rats (n=64) were randomised into control (KN), LPS (KL), probiotic + LPS (KL-Pr), and sequential probiotic + LPS + probiotic (KPR-7L) groups. Microencapsulated L. plantarum IS-10506 (2.86×1010 cfu/day) was administered via a gastric tube once daily for up to 7 days, and LPS (250 ng/kg body weight) was administered via a gastric tube on the first day of the experiment to all but the KN group. On day 3, 4, 6, and 7, four rats per group were sacrificed, and Lgr5, Bmi1, extracellular signal-regulated kinase (ERK), and β-catenin expression in the ileum was assessed by immunohistochemistry. LPS treatment reduced Lgr5 (P≤0.05) and Bmi1 (P=0.000) levels in intestinal epithelial cells, whereas probiotic treatment increased levels of Lgr5 (KPR-7L, P=0.008) and Bmi1 (KL-Pr, P=0.008; and KPR-7L, P=0.000). Lgr5 expression was upregulated in the KL-Pr group on day 3, 4, 6, and 7 (P=0.056). Additionally, ERK levels were elevated in Bmi1- and Lgr5-expressing cells in rats treated with probiotics (KL-Pr and KPR-7L), whereas β-catenin levels were increased in Lgr5-expressing cells from KPR-7L rats and in Bmi1-expressing cells from KL-Pr and KPR-7L rats on day 3 and 4. These results demonstrated that the probiotic L. plantarum IS-10506 activated intestinal stem cells to counter inflammation and might be useful for maintaining intestinal health, especially when used as a prophylactic agent.

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