Integrated in Vitro and in Silico Modelling Delineates the Molecular Effects of a Symbiotic Regimen on Colorectal Cancer-Derived Cells

Greenhalgh, Kacy; Ramiro-García, Javier; Heinken, Almut; Ullmann, Pit; Bintener, Tamara Jean Rita; Pacheco, Maria Pires; Bagińska, Joanna; Shah, Pranjul Jaykumar; Frachet, Audrey; Halder, Rashi; Fritz, Joëlle V.; Sauter, Thomas; Thiele, Ines; Haan, Serge; Letellier, Elisabeth; Wilmes, Paul

doi:10.2139/ssrn.3287784

Cited by 2 publications

(2 citation statements)

References 55 publications

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“…Samples were homogenized at 4 500 × g for 30 sec at 10 °C (Precellys24 Homogenizer) and centrifuged for 5 min at 21 000 x g and 4 °C. Further processing of the homogenate and measurements of SCFAs were then performed using high-performance liquid chromatography (HPLC), as previously described 15 .…”

Section: Intestinal Fatty Acid Analysismentioning

confidence: 99%

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium

Neumann

Steimle

Grant

et al. 2021

Preprint

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The change of dietary habits in Western societies, including reduced consumption of fiber, is linked to alterations in gut microbial ecology. Nevertheless, mechanistic connections between diet-induced microbiota changes that affect colonization resistance and enteric pathogen susceptibility are still emerging. We sought to investigate how a diet devoid of soluble plant fibers impacts the structure and function of a conventional gut microbiota in specific-pathogen-free (SPF) mice and how such changes alter susceptibility to a rodent enteric pathogen. We show that absence of dietary fiber intake leads to shifts in the abundances of specific taxa, microbiome-mediated erosion of the colonic mucus barrier, a reduction of intestinal barrier-promoting short-chain fatty acids, and increases in markers of mucosal barrier integrity disruption. Importantly, our results highlight that these low fiber diet-induced changes in the gut microbial ecology collectively contribute to a lethal colitis by the mucosal pathogen Citrobacter rodentium, which is used as a mouse model for enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively). Our study indicates that modern, low-fiber Western diets might make individuals more prone to infection by enteric pathogens via the disruption of mucosal barrier integrity by diet-driven changes in the gut microbiota, illustrating possible implications for EPEC and EHEC infections.

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Section: Intestinal Fatty Acid Analysismentioning

confidence: 99%

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium

Neumann

Steimle

Grant

et al. 2021

Preprint

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show abstract

“…25 However, the organoids lack fully organized crypt microenvironments, as well as accessible luminal regions that can be easily manipulated or assayed. 26 Among a variety of primary cell-derived microdevices, [27][28][29] in vitro intestinal crypt arrays have emerged as a promising technology for understanding intestinal homeostasis and drivers of stem cell fate decisions. [30][31][32][33] With this platform, primary intestinal cells from clinical biopsies can be cultured on a shaped hydrogel scaffold formed to mimic the crypt architecture and density observed in vivo.…”

Section: Introductionmentioning

confidence: 99%

Photopatterned Membranes and Chemical Gradients Enable Scalable Phenotypic Organization of Primary Human Colon Epithelial Models

Hinman¹,

Wang²,

Allbritton³

2019

Preprint

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Biochemical gradients across the intestinal epithelium play a major role in governing intestinal stem cell compartmentalization, differentiation dynamics, and organ-level self-renewal. Advances in primary cell-derived in vitro models, in which a full suite of stem and differentiated cell types are present, have vastly accelerated our understanding of intestinal homeostasis and disease. However, scalable platforms that recapitulate the architecture and gradients present in vivo are absent. We present a platform in which individually addressable arrays of chemical gradients along the crypt long axis can be generated, enabling scalable culture of in vitro colonic epithelial replicas. The platform utilizes standardized well plate spacing, maintains access to basal and luminal compartments, and relies on a photopatterned porous membrane to act as diffusion windows while supporting the in vitro crypts. Simultaneous fabrication of 3,875 crypts over a single membrane was developed. Growth factor gradients were modelled and then experimentally optimized to promote long-term health and self-renewal of the crypts which were assayed in situ by confocal fluorescence microscopy. The cultured in vitro crypt arrays successfully recapitulated the architecture, stem/proliferative and differentiated cell compartmentalization, and luminal-to-basal polarity observed in vivo. Furthermore, known signaling regulators produced measurable and predictable effects on the proliferative and differentiated cell compartments. This platform is readily adaptable to the screening of tissue from individual patients to assay the impact of food and bacterial metabolites and/or drugs on colonic crypt dynamics. Importantly, the cassette is compatible with a wide range of sensing/detection modalities, and the developed fabrication methods should find applications for other cell and tissue types.

show abstract

Integrated in Vitro and in Silico Modelling Delineates the Molecular Effects of a Symbiotic Regimen on Colorectal Cancer-Derived Cells

Cited by 2 publications

References 55 publications

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium

Deprivation of dietary fiber in specific-pathogen-free mice promotes susceptibility to the intestinal mucosal pathogen Citrobacter rodentium

Photopatterned Membranes and Chemical Gradients Enable Scalable Phenotypic Organization of Primary Human Colon Epithelial Models

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