Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

Nutrient sensing triggers responses by the gut-brain axis modulating hormone release, feeding behavior and metabolism that become dysregulated in metabolic syndrome and some cancers. Except for absorptive enterocytes and secretory enteroendocrine cells, the ability of many intestinal cell types to sense nutrients is still unknown; hence we hypothesized that progenitor stem cells (intestinal stem cells, ISC) possess nutrient sensing ability inherited by progenies during differentiation. We directed via modulators of Wnt and Notch signaling differentiation of precursor mouse intestinal crypts into specialized organoids each containing ISC, enterocyte, goblet, or Paneth cells at relative proportions much higher than in situ as determined by mRNA expression and immunocytochemistry of cell type biomarkers. We identified nutrient sensing cell type(s) by increased expression of fructolytic genes in response to a fructose challenge. Organoids comprised primarily of enterocytes, Paneth, or goblet, but not ISC, cells responded specifically to fructose without affecting nonfructolytic genes. Sensing was independent of Wnt and Notch modulators and of glucose concentrations in the medium but required fructose absorption and metabolism. More mature enterocyte- and goblet-enriched organoids exhibited stronger fructose responses. Remarkably, enterocyte organoids, upon forced dedifferentiation to reacquire ISC characteristics, exhibited a markedly extended lifespan and retained fructose sensing ability, mimicking responses of some dedifferentiated cancer cells. Using an innovative approach, we discovered that nutrient sensing is likely repressed in progenitor ISCs then irreversibly derepressed during specification into sensing-competent absorptive or secretory lineages, the surprising capacity of Paneth and goblet cells to detect fructose, and the important role of differentiation in modulating nutrient sensing. NEW & NOTEWORTHY Small intestinal stem cells differentiate into several cell types transiently populating the villi. We used specialized organoid cultures each comprised of a single cell type to demonstrate that 1) differentiation seems required for nutrient sensing, 2) secretory goblet and Paneth cells along with enterocytes sense fructose, suggesting that sensing is acquired after differentiation is triggered but before divergence between absorptive and secretory lineages, and 3) forcibly dedifferentiated enterocytes exhibit fructose sensing and lifespan extension.

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Nutrient sensing by absorptive and secretory progenies of small intestinal stem cells

The FASEB Journal ◽

10.1096/fasebj.31.1_supplement.1090.1 ◽

2017 ◽

Vol 31 (S1) ◽

Author(s):

Kunihiro Kishida ◽

Sarah Pearce ◽

Shiyan Yu ◽

Nan Gao ◽

Ronaldo Ferraris

Keyword(s):

Stem Cells ◽

Nutrient Sensing ◽

Intestinal Stem Cells ◽

Small Intestinal

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Mouse telomerase reverse transcriptase (mTert) expression marks slowly cycling intestinal stem cells

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1013004108 ◽

2010 ◽

Vol 108 (1) ◽

pp. 179-184 ◽

Cited By ~ 337

Author(s):

Robert K. Montgomery ◽

Diana L. Carlone ◽

Camilla A. Richmond ◽

Loredana Farilla ◽

Mariette E. G. Kranendonk ◽

...

Keyword(s):

Stem Cells ◽

Reverse Transcriptase ◽

Tissue Injury ◽

Cell Types ◽

Lineage Tracing ◽

Intestinal Stem Cells ◽

Intestinal Cell ◽

Stem Cell Population ◽

Telomerase Reverse Transcriptase

The intestinal epithelium is maintained by a population of rapidly cycling (Lgr5+) intestinal stem cells (ISCs). It has been postulated, however, that slowly cycling ISCs must also be present in the intestine to protect the genome from accumulating deleterious mutations and to allow for a response to tissue injury. Here, we identify a subpopulation of slowly cycling ISCs marked by mouse telomerase reverse transcriptase (mTert) expression that can give rise to Lgr5+ cells. mTert-expressing cells distribute in a pattern along the crypt–villus axis similar to long-term label-retaining cells (LRCs) and are resistant to tissue injury. Lineage-tracing studies demonstrate that mTert+ cells give rise to all differentiated intestinal cell types, persist long term, and contribute to the regenerative response following injury. Consistent with other highly regenerative tissues, our results demonstrate that a slowly cycling stem cell population exists within the intestine.

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A novel in vitro survival assay of small intestinal stem cells after exposure to ionizing radiation

Journal of Radiation Research ◽

10.1093/jrr/rrt123 ◽

2014 ◽

Vol 55 (2) ◽

pp. 381-390 ◽

Cited By ~ 11

Author(s):

Motohiro Yamauchi ◽

Kensuke Otsuka ◽

Hisayoshi Kondo ◽

Nobuyuki Hamada ◽

Masanori Tomita ◽

...

Keyword(s):

Stem Cells ◽

Ionizing Radiation ◽

Intestinal Stem Cells ◽

Survival Assay ◽

Small Intestinal ◽

In Vitro Survival

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Abstract 17093: Human Neonatal Cardiac Mesenchymal Stem Cells Demonstrates Superior Cardiac Regenerative Abilities Compared to Other Stem Cells

Circulation ◽

10.1161/circ.142.suppl_3.17093 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

Muthukumar Gunasekaran ◽

Rachana Mishra ◽

Progyaparamita Saha ◽

Xuebin Fu ◽

Mohamed Abdullah ◽

...

Keyword(s):

Myocardial Infarction ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Inflammatory Cytokines ◽

Cell Types ◽

Wild Type ◽

Cell Type ◽

Anti Inflammatory ◽

Stem Cell Type

Stem cells transplantation is being explored as an effective therapy for heart diseases. However, majority of stem cell therapies for adult patients with myocardial infarction (MI) had mixed and inconsistent results implying chronological age may influence the effectiveness of regenerative therapies. Therefore, herein, we performed a head-to-head comparison between different, well-studied stem cell types to identify the superior regenerative cell type using rodent MI model.After our standard characterization for each stem cell type (FACS for cell surface markers), 1 million neonatal Cardiac Mesenchymal Stem cells (nMSCs), adult MSCs (aMSCs), adult derived cardiosphere derived cells (aCDCs), umbilical cord derived cells (UCBCs), Bone Marrow derived Mesenchymal Stem cells (BM-MSCs), or cell-free Iscove Modified Dulbecco Medium (IMDM as placebo control) were injected into athymic rat myocardial infarct model. Although all the tested groups significantly improved ejection fraction, nMSCs outperformed other stem cells in cardiac functional recovery. Additionally, nMSCs also showed significant increased cardiac functional recovery compared to aMSCs in wild type rat MI model. Mason trichrome staining with heart sections revealed that decreased fibrosis was evident on nMSCs injection compared to aMSCs in both athymic and wild type rat MI model. Myocardial sections from rats received nMSCs showed significantly reduced M1 macrophages (inflammatory) and increased M2 macrophages (anti-inflammatory) compared with sections from rats having received aMSCs and IMDM control. Pro and anti-inflammatory cytokines analyzed on sera collected on day 2 and 7 revealed that anti-inflammatory cytokine (IL10) was significantly increased and inflammatory cytokines (IL4 and IL12) reduced in nMSCs compared to aMSCs transplanted MI rat model.In conclusion, nMSCs demonstrated superior functional abilities, reduced fibrosis, inflammatory cells and cytokines compared to all the other cell types and with aMSCs demonstrating that nMSCs is an ideal stem cell type for therapeutic application in myocardial infarction.

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Heterogeneity in histone 2B-green fluorescent protein-retaining putative small intestinal stem cells at cell position 4 and their absence in the colon

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00080.2012 ◽

2012 ◽

Vol 303 (11) ◽

pp. G1188-G1201 ◽

Cited By ~ 20

Author(s):

Kevin R. Hughes ◽

Ricardo M. C. Gândara ◽

Tanvi Javkar ◽

Fred Sablitzky ◽

Hanno Hock ◽

...

Keyword(s):

Stem Cells ◽

Green Fluorescent Protein ◽

Fluorescent Protein ◽

Paneth Cells ◽

Epithelial Stem Cells ◽

Epithelial Regeneration ◽

Cell Position ◽

Small Intestinal ◽

Green Fluorescent ◽

Dose Radiation

Stem cells have been identified in two locations in small intestinal crypts; those intercalated between Paneth cells and another population (which retains DNA label) are located above the Paneth cell zone, at cell position 4. Because of disadvantages associated with the use of DNA label, doxycycline-induced transient transgenic expression of histone 2B (H2B)-green fluorescent protein (GFP) was investigated. H2B-GFP-retaining putative stem cells were consistently seen, with a peak at cell position 4, over chase periods of up to 112 days. After a 28-day chase, a subpopulation of the H2B-GFP-retaining cells was cycling, but the slow cycling status of the majority was illustrated by lack of expression of pHistone H3 and Ki67. Although some H2B-GFP-retaining cells were sensitive to low-dose radiation, the majority was resistant to low- and high-dose radiation-induced cell death, and a proportion of the surviving cells proliferated during subsequent epithelial regeneration. Long-term retention of H2B-GFP in a subpopulation of small intestinal Paneth cells was also seen, implying that they are long lived. In contrast to the small intestine, H2B-GFP-retaining epithelial cells were not seen in the colon from 28-day chase onward. This implies important differences in stem cell function between these two regions of the gastrointestinal tract, which may have implications for region-specific susceptibility to diseases (such as cancer and ulcerative colitis), in which epithelial stem cells and their progeny are involved.

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Cell-Type–Specific, Ketohexokinase-Dependent Induction by Fructose of Lipogenic Gene Expression in Mouse Small Intestine

Journal of Nutrition ◽

10.1093/jn/nxaa113 ◽

2020 ◽

Vol 150 (7) ◽

pp. 1722-1730 ◽

Cited By ~ 3

Author(s):

Arwa Al-Jawadi ◽

Chirag R Patel ◽

Reilly J Shiarella ◽

Emmanuellie Romelus ◽

Madelyn Auvinen ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Metabolic Diseases ◽

Free Access ◽

Wild Type ◽

Cell Type ◽

Lipogenic Gene ◽

Lipogenic Genes ◽

Lipogenic Gene Expression ◽

Small Intestinal

ABSTRACT Background High intakes of fructose are associated with metabolic diseases, including hypertriglyceridemia and intestinal tumor growth. Although small intestinal epithelia consist of many different cell types, express lipogenic genes, and convert dietary fructose to fatty acids, there is no information on the identity of the cell type(s) mediating this conversion and on the effects of fructose on lipogenic gene expression. Objectives We hypothesized that fructose regulates the intestinal expression of genes involved in lipid and apolipoprotein synthesis, that regulation depends on the fructose transporter solute carrier family 2 member a5 [Slc2a5 (glucose transporter 5)] and on ketohexokinase (Khk), and that regulation occurs only in enterocytes. Methods We compared lipogenic gene expression among different organs from wild-type adult male C57BL mice consuming a standard vivarium nonpurified diet. We then gavaged twice daily for 2.5 d fructose or glucose solutions (15%, 0.3 mL per mouse) into wild-type, Slc2a5-knockout (KO), and Khk-KO mice with free access to the nonpurified diet and determined expression of representative lipogenic genes. Finally, from mice fed the nonpurified diet, we made organoids highly enriched in enterocyte, goblet, Paneth, or stem cells and then incubated them overnight in 10 mM fructose or glucose. Results Most lipogenic genes were significantly expressed in the intestine relative to the kidney, liver, lung, and skeletal muscle. In vivo expression of Srebf1, Acaca, Fasn, Scd1, Dgat1, Gk, Apoa4, and Apob mRNA and of Scd1 protein increased (P < 0.05) by 3- to 20-fold in wild-type, but not in Slc2a5-KO and Khk-KO, mice gavaged with fructose. In vitro, Slc2a5- and Khk-dependent, fructose-induced increases, which ranged from 1.5- to 4-fold (P < 0.05), in mRNA concentrations of all these genes were observed only in organoids enriched in enterocytes. Conclusions Fructose specifically stimulates expression of mouse small intestinal genes for lipid and apolipoprotein synthesis. Secretory and stem cells seem incapable of transport- and metabolism-dependent lipogenesis, occurring only in absorptive enterocytes.

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Mo1835 Investigation of Slow Cycling Stem Cells and Paneth Cells in T. Spiralis-Induced Small Intestinal Inflammation

Gastroenterology ◽

10.1016/s0016-5085(15)32464-1 ◽

2015 ◽

Vol 148 (4) ◽

pp. S-722

Author(s):

Tanvi Javkar ◽

Fred Sablitzky ◽

Yashwant R. Mahida

Keyword(s):

Stem Cells ◽

Intestinal Inflammation ◽

Paneth Cells ◽

Small Intestinal ◽

Slow Cycling

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Chemical compound-based direct reprogramming for future clinical applications

Bioscience Reports ◽

10.1042/bsr20171650 ◽

2018 ◽

Vol 38 (3) ◽

Cited By ~ 10

Author(s):

Yukimasa Takeda ◽

Yoshinori Harada ◽

Toshikazu Yoshikawa ◽

Ping Dai

Keyword(s):

Stem Cells ◽

Transcription Factors ◽

Chemical Compound ◽

Chemical Compounds ◽

Cell Types ◽

Direct Reprogramming ◽

Cell Type ◽

Promising Alternative ◽

Cell Functions ◽

Transplantation Therapy

Recent studies have revealed that a combination of chemical compounds enables direct reprogramming from one somatic cell type into another without the use of transgenes by regulating cellular signaling pathways and epigenetic modifications. The generation of induced pluripotent stem (iPS) cells generally requires virus vector-mediated expression of multiple transcription factors, which might disrupt genomic integrity and proper cell functions. The direct reprogramming is a promising alternative to rapidly prepare different cell types by bypassing the pluripotent state. Because the strategy also depends on forced expression of exogenous lineage-specific transcription factors, the direct reprogramming in a chemical compound-based manner is an ideal approach to further reduce the risk for tumorigenesis. So far, a number of reported research efforts have revealed that combinations of chemical compounds and cell-type specific medium transdifferentiate somatic cells into desired cell types including neuronal cells, glial cells, neural stem cells, brown adipocytes, cardiomyocytes, somatic progenitor cells, and pluripotent stem cells. These desired cells rapidly converted from patient-derived autologous fibroblasts can be applied for their own transplantation therapy to avoid immune rejection. However, complete chemical compound-induced conversions remain challenging particularly in adult human-derived fibroblasts compared with mouse embryonic fibroblasts (MEFs). This review summarizes up-to-date progress in each specific cell type and discusses prospects for future clinical application toward cell transplantation therapy.

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Effects of Aging and TGF-Beta 3 on Chondrocyte and Mesenchymal Stem Cell Matrix Formation

ASME 2010 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2010-19517 ◽

2010 ◽

Author(s):

Isaac E. Erickson ◽

Steven C. van Veen ◽

Swarnali Sengupta ◽

Sydney R. Kestle ◽

Jason A. Burdick ◽

...

Keyword(s):

Stem Cells ◽

Articular Chondrocytes ◽

Cell Types ◽

Matrix Proteins ◽

Cell Type ◽

Cell Matrix ◽

Age Related ◽

Cartilage Restoration ◽

Effects Of Aging

Articular cartilage pathology is common in the aged population. Numerous studies have shown that aged chondrocytes (CHs) are inferior to juvenile CHs in their ability to proliferate and produce cartilage-specific extracellular matrix proteins, potentially limiting their use in tissue engineering applications for cartilage restoration [1,2]. Mesenchymal stem cells (MSCs) are an alternative cell type that can be expanded in vitro while maintaining their ability to differentiate into cell types comparable to articular chondrocytes. However, organismal aging also influences human MSC proliferation [3,4] and multi-potential differentiation [5], though for chondrogenesis these findings are mixed, with some suggesting that aged progenitor cells retain their chondrogenic capacity [6]. The objective of this study was to assess age related differences in donor-matched CH and MSC potential for chondrogenic repair. In addition, the effects of the chondrogenic growth factor TGF-β3 on CHs and MSCs were evaluated.

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