Analysis of Wnt signalling dynamics during colon crypt development in 3D culture

Tan, Chin Wee; Hirokawa, Yumiko; Burgess, Antony W.

doi:10.1038/srep11036

Cited by 14 publications

(18 citation statements)

References 38 publications

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“…The detailed image acquisition and analysis procedures for acquiring images in 96‐well plates have been described previously (Hirokawa et al , ; Tan et al , ). In this experiment, 384‐well plates (Corning #3985) were used.…”

Section: Methodsmentioning

confidence: 99%

Repurposing the selective estrogen receptor modulator bazedoxifene to suppress gastrointestinal cancer growth

et al. 2019

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Excessive signaling through gp130, the shared receptor for the interleukin ( IL )6 family of cytokines, is a common hallmark in solid malignancies and promotes their progression. Here, we established the in vivo utility of bazedoxifene, a steroid analog clinically approved for the treatment of osteoporosis, to suppress gp130‐dependent tumor growth of the gastrointestinal epithelium. Bazedoxifene administration reduced gastric tumor burden in gp130 Y757F mice, where tumors arise exclusively through excessive gp130/ STAT 3 signaling in response to the IL 6 family cytokine IL 11. Likewise, in mouse models of sporadic colon and intestinal cancers, which arise from oncogenic mutations in the tumor suppressor gene Apc and the associated β‐catenin/canonical WNT pathway, bazedoxifene treatment reduces tumor burden. Consistent with the proposed orthogonal tumor‐promoting activity of IL 11‐dependent gp130/ STAT 3 signaling, tumors of bazedoxifene ‐treated Apc ‐mutant mice retain excessive nuclear accumulation of β‐catenin and aberrant WNT pathway activation. Likewise, bazedoxifene treatment of human colon cancer cells harboring mutant APC did not reduce aberrant canonical WNT signaling, but suppressed IL 11‐dependent STAT 3 signaling. Our findings provide compelling proof of concept to support the repurposing of bazedoxifene for the treatment of gastrointestinal cancers in which IL 11 plays a tumor‐promoting role.

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Section: Methodsmentioning

confidence: 99%

Repurposing the selective estrogen receptor modulator bazedoxifene to suppress gastrointestinal cancer growth

et al. 2019

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“…Intestinal crypt isolation was performed as previously described by Tan et al . 10 , with few modifications. In brief, the distal half of the colon from C57BL/6 WT mice, excluding the cecum and the most proximal portion, was harvested and transferred to a petri dish on ice; then, the sample was flushed with ice-cold phosphate-buffered saline (PBS) and cut open longitudinally.…”

Section: Methodsmentioning

confidence: 99%

Cytoglobin affects tumorigenesis and the expression of ulcerative colitis-associated genes under chemically induced colitis in mice

Yassin

Kissow

Vainer

et al. 2018

Sci Rep

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Cytoglobin (Cygb) is a member of the hemoglobin family and is thought to protect against cellular hypoxia and oxidative stress. These functions may be particularly important in inflammation-induced cancer, e.g., in patients with ulcerative colitis (UC). In this study, we investigated the development of inflammation and tumors in a murine model of inflammation-induced colorectal cancer using a combined treatment of azoxymethane and dextran sulfate sodium. A bioinformatics analysis of genome-wide expression data revealed increased colonic inflammation at the molecular level accompanied by enhanced macroscopic tumor development in Cygb-deficient mice. Moreover, the expression of the UC-associated gene neurexophilin and PC-esterase domain family member 4 (Nxpe4) depended on the presence of Cygb in the inflamed colonic mucosa. Compared to wild type mice, RT-qPCR confirmed a 14-fold (p = 0.0003) decrease in Nxpe4 expression in the inflamed colonic mucosa from Cygb-deficient mice. An analysis of Cygb protein expression suggested that Cygb is expressed in fibroblast-like cells surrounding the colonic crypts. Histological examinations of early induced lesions suggested that the effect of Cygb is primarily at the level of tumor promotion. In conclusion, in this model, Cygb primarily seemed to inhibit the development of established microadenomas.

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“…One possible explanation for this extended stability is that with every medium exchange, culture debris is removed and the organoids in the microwells are in direct contact with fresh growth factors and other cytokines contained in the medium, whereas nutrients have to diffuse through the gel for organoids embedded in Matrigel domes and metabolic waste products are more likely to remain. [43] Alternatively, organoids grown in microwells may be exposed to higher and more equally distributed oxygen concentrations to facilitate growth and viability, as demonstrated previously for organoids in suspension cultures compared to ones grown embedded at different locations and different depths within dense gels. Okkelman et al [37] demonstrated that organoids randomly distributed within Matrigel domes are not equally exposed to oxygen, imposing differences between organoids located close to the surface and deeper in the matrix.…”

Section: Organoids In Microwells Show Lower Cell Death Ratesmentioning

confidence: 99%

Intestinal Organoid Culture in Polymer Film‐Based Microwell Arrays

et al. 2020

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compartment occupied by continuously dividing cells at the bottom, which after differentiation migrate upward toward the villus. In the villus, six different mature cell types are found, including secretory cells, such as goblet, Paneth, enteroendocrine, and tuft cells, and also absorptive cells, such as enterocytes and microfold cells. [1-3] The intestinal epithelium serves two main functions, nutrient uptake and protection against harmful substances and pathogens. Τhe presence of strong apical-basolateral compartmentalization is of critical importance for proper intestinal function. [3-7] Despite various advances in cell and tissue culture technologies [8-11] and organ-on-chip models [12] that have been used to study intestinal physiology over the past years, numerous morphological and functional aspects remain unknown. Miniaturized 3D, multicellular, stem cell-derived constructs that mimic in vivo tissue, called organoids, along with organ-on-chip systems represent culture systems able to recapitulate the complexity of an organ closer than any previously utilized technique. [13,14] Organoids have high self-organization capacity and hold great potential as a research tool to explore unknown aspects of organ development, tissue regeneration, disease pathology, cell biology, and as drug-screening platforms. In the past few years, numerous protocols have been described to grow organoids that resemble various organs, such as liver, brain, intestine, and lung. [15-21] Intestinal organoid culture is a relatively simple system, that typically involves the embedding of small multicellular fragments (containing LGR5 + cells) in Matrigel, which serves as an extracellular matrix mimic, supplemented with the right cocktail of growth factors. [16] This results in growing and self-sustaining intestinal organoids, which contain all the above mentioned cell types found in vivo and are organized in a crypt-villus structure that surrounds a central lumen, with strong apical-basolateral polarity. [13,22] Hence, these organoids have contributed significantly to the understanding of normal intestinal function and dysfunction in the last years. Even though organoids are a powerful tool, their use still faces numerous limitations. Bioengineering approaches hold great potential in overcoming some constraints. [23-27] More specifically, mass transport in organoids is usually restricted by their size, a limitation which researchers hope to overcome with either the use of bioreactors, microfluidic chips or integration of vascular

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Analysis of Wnt signalling dynamics during colon crypt development in 3D culture

Cited by 14 publications

References 38 publications

Repurposing the selective estrogen receptor modulator bazedoxifene to suppress gastrointestinal cancer growth

Repurposing the selective estrogen receptor modulator bazedoxifene to suppress gastrointestinal cancer growth

Cytoglobin affects tumorigenesis and the expression of ulcerative colitis-associated genes under chemically induced colitis in mice

Intestinal Organoid Culture in Polymer Film‐Based Microwell Arrays

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