32 results on '"von Furstenberg, Richard J."'
Search Results
2. A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion
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Boone, Peter G., Rochelle, Lauren K., Ginzel, Joshua D., Lubkov, Veronica, Roberts, Wendy L., Nicholls, P. J., Bock, Cheryl, Flowers, Mei Lang, von Furstenberg, Richard J., Stripp, Barry R., Agarwal, Pankaj, Borowsky, Alexander D., Cardiff, Robert D., Barak, Larry S., Caron, Marc G., Lyerly, H. Kim, and Snyder, Joshua C.
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- 2019
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3. Side population sorting separates subfractions of cycling and non-cycling intestinal stem cells
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von Furstenberg, Richard J., Buczacki, Simon J.A., Smith, Brian J., Seiler, Kristen M., Winton, Douglas J., and Henning, Susan J.
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- 2014
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4. Tissue underlying the intestinal epithelium elicits proliferation of intestinal stem cells following cytotoxic damage
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Seiler, Kristen M, Schenhals, Erica L, von Furstenberg, Richard J, Allena, Bhavya K, Smith, Brian J, Scaria, Denny, Bresler, Michele N, Dekaney, Christopher M, and Henning, Susan J
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- 2015
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5. Intestinal stem cells remain viable after prolonged tissue storage
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Fuller, Megan K., Faulk, Denver M., Sundaram, Nambirajan, Mahe, Maxime M., Stout, Kara M., von Furstenberg, Richard J., Smith, Brian J., McNaughton, Kirk K., Shroyer, Noah F., Helmrath, Michael A., and Henning, Susan J.
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- 2013
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6. Mouse Paneth cell antimicrobial function is independent of Nod2
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Shanahan, Michael T, Carroll, Ian M, Grossniklaus, Emily, White, Andrew, von Furstenberg, Richard J, Barner, Roshonda, Fodor, Anthony A, Henning, Susan J, Sartor, R Balfour, and Gulati, Ajay S
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- 2014
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7. Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal
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Yan, Kelley S., Janda, Claudia Y., Chang, Junlei, Zheng, Grace X. Y., Larkin, Kathryn A., Luca, Vincent C., Chia, Luis A., Mah, Amanda T., Han, Arnold, Terry, Jessica M., Ootani, Akifumi, Roelf, Kelly, Lee, Mark, Yuan, Jenny, Li, Xiao, Bolen, Christopher R., Wilhelmy, Julie, Davies, Paige S., Ueno, Hiroo, von Furstenberg, Richard J., Belgrader, Phillip, Ziraldo, Solongo B., Ordonez, Heather, Henning, Susan J., Wong, Melissa H., Snyder, Michael P., Weissman, Irving L., Hsueh, Aaron J., Mikkelsen, Tarjei S., Garcia, K. Christopher, and Kuo, Calvin J.
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Ligands (Biochemistry) -- Health aspects ,Stem cells -- Health aspects ,Environmental issues ,Science and technology ,Zoology and wildlife conservation - Abstract
Author(s): Kelley S. Yan [1, 2]; Claudia Y. Janda [3]; Junlei Chang [1]; Grace X. Y. Zheng [4]; Kathryn A. Larkin [1]; Vincent C. Luca [3]; Luis A. Chia [1]; [...]
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- 2017
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8. Rosiglitazone attenuates suppression of RXRα-dependent gene expression in inflamed liver
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Ghose, Romi, Mulder, Jaap, von Furstenberg, Richard J., Thevananther, Sundararajah, Kuipers, Folkert, and Karpen, Saul J.
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- 2007
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9. Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal
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Belgrader, Phillip, Ueno, Hiroo, Yuan, Jenny, Henning, Susan J., Lee, Mark, Terry, Jessica M., Wong, Melissa H., Ziraldo, Solongo B., Han, Arnold, Li, Xiao, Larkin, Kathryn A., Luca, Vincent C., Ootani, Akifumi, Von Furstenberg, Richard J., Snyder, Michael P., Ordonez, Heather, Kuo, Calvin J., Weissman, Irving L., Chang, Junlei, Hsueh, Aaron J., Yan, Kelley S., Chia, Luis A., Zheng, Grace X. Y., Wilhelmy, Julie, Roelf, Kelly, Garcia, K. Christopher, Janda, Claudia Y., Mikkelsen, Tarjei S., Davies, Paige S., Bolen, Christopher R., and Mah, Amanda T.
- Abstract
The canonical Wnt/β-catenin signaling pathway governs diverse developmental, homeostatic and pathologic processes. Palmitoylated Wnt ligands engage cell surface Frizzled (Fzd) receptors and Lrp5/6 co-receptors enabling β-catenin nuclear translocation and Tcf/Lef-dependent gene transactivation1–3. Mutations in Wnt downstream signaling components have revealed diverse functions presumptively attributed to Wnt ligands themselves, although direct attribution remains elusive, as complicated by redundancy between 19 mammalian Wnts and 10 Fzds1 and Wnt hydrophobicity2,3. For example, individual Wnt ligand mutations have not revealed homeostatic phenotypes in the intestinal epithelium4, an archetypal canonical Wnt pathway-dependent rapidly self-renewing tissue whose regeneration is fueled by proliferative crypt Lgr5+ intestinal stem cells (ISCs)5–9. R-spondin ligands (Rspo1–4) engage distinct Lgr4-6 and Rnf43/Znrf3 receptor classes10–13, markedly potentiate canonical Wnt/β-catenin signaling and induce intestinal organoid growth in vitro and Lgr5+ ISCs in vivo8,14–17. However, the interchangeability, functional cooperation and relative contributions of Wnt versus Rspo ligands to in vivo canonical Wnt signaling and ISC biology remain unknown. Here, we deconstructed functional roles of Wnt versus Rspo ligands in the intestinal crypt stem cell niche. We demonstrate that the default fate of Lgr5+ ISCs is lineage commitment, escape from which requires both Rspo and Wnt ligands. However, gain-of-function studies using Rspo versus a novel non-lipidated Wnt analog reveal qualitatively distinct, non-interchangeable roles for these ligands in ISCs. Wnts are insufficient to induce Lgr5+ ISC self-renewal, but rather confer a basal competency by maintaining Rspo receptor expression that enables Rspo to actively drive and specify the extent of stem cell expansion. This functionally non-equivalent yet cooperative interplay between Wnt and Rspo ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precision control of tissue regeneration.
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- 2017
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10. Prior tonsillectomy is associated with an increased risk of esophageal adenocarcinoma.
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Garman, Katherine S., Ajayi, Teminioluwa A., Boutte, Harold J., Chiu, Shih-Ting, von Furstenberg, Richard J., Lloyd, Benjamin R., Zhang, Cecelia, Onaitis, Mark W., Chow, Shein-Chung, and McCall, Shannon J.
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TONSILLECTOMY ,BARRETT'S esophagus ,SURGICAL indications ,THORACIC surgery ,LOGISTIC regression analysis ,GLEASON grading system ,ODDS ratio ,STAPLERS (Surgery) - Abstract
Background: Esophageal cancer is a deadly cancer with 5-year survival <20%. Although multiple risk factors for esophageal adenocarcinoma (EAC) including obesity, GERD and smoking have been identified, these risk factors do not fully explain the rising incidence of EAC. In this study, we evaluated the association between prior history of tonsillectomy and EAC. Our goal was to determine whether tonsillectomies were more frequent in patients with EAC (cases) than in our thoracic surgery controls. Methods: Cases included 452 esophagectomy cases, including 396 with EAC and 56 who underwent esophagectomy for Barrett's esophagus (BE) with high grade dysplasia (HGD). 1,102 thoracic surgery patients with surgical indications other than dysplastic BE or esophageal cancer represented the controls for our analysis. The association of tonsillectomy and HGD/EAC were primarily evaluated by using univariate tests and then verified by logistic regression analysis. Baseline demographics, medical history, and thoracic surgery controls were compared by using χ
2 tests or 95% CIs. Significant risk factors were considered as covariates in the multivariate models while evaluating the association between tonsillectomy and HGD/EAC. P-values or odds ratios were estimated with 95% confidence limits to identify significances which was more appropriate. Results: Tonsillectomy was more common in cases than controls and was found to have a significant association with esophageal cancer (19.9% vs. 12.7%; p-value = 0.0003). This significant association persisted after controlling for other known risk factors/covariates. Conclusion: A prior history of tonsillectomy was significantly associated with HGD/EAC and may represent an independent risk factor for the development of EAC. However, the underlying biology driving this association remains unclear. [ABSTRACT FROM AUTHOR]- Published
- 2020
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11. Plasmonic nanobiosensors for detection of microRNA cancer biomarkers in clinical samples.
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Crawford, Bridget M., Wang, Hsin-Neng, Stolarchuk, Christina, von Furstenberg, Richard J., Strobbia, Pietro, Zhang, Dadong, Qin, Xiaodi, Owzar, Kouros, Garman, Katherine S., and Vo-Dinh, Tuan
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BARRETT'S esophagus ,EARLY diagnosis ,MICRORNA ,ESOPHAGEAL cancer ,RAMAN scattering - Abstract
MicroRNAs (miRNAs) play an important role in the regulation of biological processes and have demonstrated great potential as biomarkers for the early detection of various diseases, including esophageal adenocarcinoma (EAC) and Barrett's esophagus (BE), the premalignant metaplasia associated with EAC. Herein, we demonstrate the direct detection of the esophageal cancer biomarker, miR-21, in RNA extracted from 17 endoscopic tissue biopsies using the nanophotonics technology our group has developed, termed the inverse molecular sentinel (iMS) nanobiosensor, with surface-enhanced Raman scattering (SERS) detection. The potential of this label-free, homogeneous biosensor for cancer diagnosis without the need for target amplification was demonstrated by discriminating esophageal cancer and Barrett's esophagus from normal tissue with notable diagnostic accuracy. This work establishes the potential of the iMS nanobiosensor for cancer diagnostics via miRNA detection in clinical samples without the need for target amplification, validating the potential of this assay as part of a new diagnostic strategy. Combining miRNA diagnostics with the nanophotonics technology will result in a paradigm shift in achieving a general molecular analysis tool that has widespread applicability for cancer research as well as detection of cancer. We anticipate further development of this technique for future use in point-of-care testing as an alternative to histopathological diagnosis as our method provides a quick result following RNA isolation, allowing for timely treatment. [ABSTRACT FROM AUTHOR]
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- 2020
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12. GI stem cells – new insights into roles in physiology and pathophysiology
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Henning, Susan J. and von Furstenberg, Richard J.
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inorganic chemicals ,Intestines ,Stem Cells ,fungi ,Cell Culture Techniques ,Animals ,Humans ,Special section reviews: GI stem cells ‐ new insights into roles in physiology and pathophysiology ,Intestinal Mucosa ,digestive system - Abstract
This overview gives a brief historical summary of key discoveries regarding stem cells of the small intestine. The current concept is that there are two pools of intestinal stem cells (ISCs): an actively cycling pool that is marked by Lgr5, is relatively homogeneous and is responsible for daily turnover of the epithelium; and a slowly cycling or quiescent pool that functions as reserve ISCs. The latter pool appears to be quite heterogeneous and may include partially differentiated epithelial lineages that can reacquire stem cell characteristics following injury to the intestine. Markers and methods of isolation for active and quiescent ISC populations are described as well as the numerous important advances that have been made in approaches to the in vitro culture of ISCs and crypts. Factors regulating ISC biology are briefly summarized and both known and unknown aspects of the ISC niche are discussed. Although most of our current knowledge regarding ISC physiology and pathophysiology has come from studies with mice, recent work with human tissue highlights the potential translational applications arising from this field of research. Many of these topics are further elaborated in the following articles.
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- 2016
13. The enteric microbiota regulates jejunal Paneth cell number and function without impacting intestinal stem cells.
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Schoenborn, Alexi A., von Furstenberg, Richard J., Valsaraj, Smrithi, Hussain, Farah S., Stein, Molly, Shanahan, Michael T., Henning, Susan J., and Gulati, Ajay S.
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- 2019
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14. Side population sorting separates subfractions of cycling and non-cycling intestinal stem cells
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Seiler, Kristen M., Henning, Susan J., von Furstenberg, Richard J., and Winton, Douglas J.
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fungi ,digestive system - Abstract
Isolation of quiescent intestinal stem cells (ISCs) has remained a challenge.By side population (SP) sorting we show cycling ISCs to be in the upper SP.The lower SP is non-cycling and has markers of the quiescent ISCs.In culture both upper and lower SP cells yield all 4 intestinal lineages.Evidence for a role of lower SP cells in repair of intestinal damage is discussed.We report here that side population (SP) sorting allows for the simultaneous isolation of two intestinal stem cell (ISC) subsets from wild-type (WT) mice which are phenotypically different and represent cycling and non-cycling pools of cells. Following 5-ethynyl-2′-deoxyuridine (EdU) injection, in the upper side population (USP) the percentage of EdU + was 36% showing this fraction to be highly proliferative. In the lower side population (LSP), only 0.4% of cells were EdU +, indicating this fraction to be predominantly non-cycling. Using Lgr5-EGFP mice, we show that Lgr5-EGFPhi cells, representing actively cycling ISCs, are essentially exclusive to the USP. In contrast, using histone 2B-YFP mice, SP analysis revealed YFP label retaining cells (LRCs) in both the USP and the LSP. Correspondingly, evaluation of the SP fractions for mRNA markers by qRT-PCR showed that the USP was enriched in transcripts associated with both quiescent and active ISCs. In contrast, the LSP expressed mRNA markers of quiescent ISCs while being de-enriched for those of the active ISC. Both the USP and LSP are capable of generating enteroids in culture which include the four intestinal lineages. We conclude that sorting of USP and LSP fractions represents a novel isolation of cycling and non-cycling ISCs from WT mice.Figure optionsDownload full-size imageDownload high-quality image (243 K)Download as PowerPoint slide
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- 2014
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15. Su1762 - Esophageal Submucosal Glands as a Source of Progenitor Cells Capable of Differentiating Into Squamous or Barrett's-Like Columnar Cells
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Iich, Elhadi, von Furstenberg, Richard J., Garman, Katherine S., Duong, Cuong, Clemons, Nicholas J., and Phillip, Wayne A.
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- 2018
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16. 1075 - Increased Gastrin Responsiveness after Injury in Esophageal Submucosal Glands
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Martinez-Uribe, Omar, von Furstenberg, Richard J., Su, Zuowei, Hussain, Farah S., McCall, Shannon J., and Garman, Katherine S.
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- 2018
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17. Ductular and proliferative response of esophageal submucosal glands in a porcine model of esophageal injury and repair.
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Krüger, Leandi, Gonzalez, Liara M., Pridgen, Tiffany A., McCall, Shannon J., von Furstenberg, Richard J., Harnden, Ivan, Carnighan, Gwendolyn E., Cox, Abigail M., Blikslager, Anthony T., and Garman, Katherine S.
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ESOPHAGEAL injuries ,BARRETT'S esophagus ,ADENOCARCINOMA - Abstract
Esophageal injury is a risk factor for diseases such as Barrett's esophagus (BE) and esophageal adenocarcinoma. To improve understanding of signaling pathways associated with both normal and abnormal repair, animal models are needed. Traditional rodent models of esophageal repair are limited by the absence of esophageal submucosal glands (ESMGs), which are present in the human esophagus. Previously, we identified acinar ductal metaplasia in human ESMGs in association with both esophageal injury and cancer. In addition, the SOX9 transcription factor has been associated with generation of columnar epithelium and the pathogenesis of BE and is present in ESMGs. To test our hypothesis that ESMGs activate after esophageal injury with an increase in proliferation, generation of a ductal phenotype, and expression of SOX9, we developed a porcine model of esophageal injury and repair using radiofrequency ablation (RFA). The porcine esophagus contains ESMGs, and RFA produces a consistent and reproducible mucosal injury in the esophagus. Here we present a temporal assessment of this model of esophageal repair. Porcine esophagus was evaluated at 0, 6, 18, 24, 48, and 72 h and 5 and 7 days following RFA and compared with control uninjured esophagus. Following RFA, ESMGs demonstrated an increase in ductal phenotype, echoing our prior studies in humans. Proliferation increased in both squamous epithelium and ESMGs postinjury with a prominent population of SOX9-positive cells in ESMGs postinjury. This model promises to be useful in future experiments evaluating mechanisms of esophageal repair. [ABSTRACT FROM AUTHOR]
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- 2017
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18. Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal.
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Yan, Kelley S., Janda, Claudia Y., Chang, Junlei, Zheng, Grace X. Y., Larkin, Kathryn A., Luca, Vincent C., Chia, Luis A., Mah, Amanda T., Han, Arnold, Terry, Jessica M., Ootani, Akifumi, Roelf, Kelly, Lee, Mark, Yuan, Jenny, Li, Xiao, Bolen, Christopher R., Wilhelmy, Julie, Davies, Paige S., Ueno, Hiroo, and von Furstenberg, Richard J.
- Abstract
The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium-an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5
+ intestinal stem cells (ISCs). R-spondin ligands (RSPO1-RSPO4) engage distinct LGR4-LGR6, RNF43 and ZNRF3 receptor classes, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5+ ISCs in vivo. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5+ ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5+ ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration. [ABSTRACT FROM AUTHOR]- Published
- 2017
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19. The Viral Mimetic Polyinosinic:Polycytidylic Acid Alters the Growth Characteristics of Small Intestinal and Colonic Crypt Cultures.
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Davies, Julie M., Santaolalla, Rebeca, von Furstenberg, Richard J., Henning, Susan J., and Abreu, Maria T.
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SMALL intestine physiology ,PATHOGENIC microorganisms ,TOLL-like receptors ,LIGANDS (Biochemistry) ,FLOW cytometry ,CELL culture - Abstract
Background & Aims: The intestinal epithelium is the first line of defense against enteric pathogens. We investigated the response of small intestinal and colonic crypt cultures to a panel of toll-like receptor ligands to assess the impact of microbial pattern recognition on epithelial growth. Methods: Primary murine jejunal enteroids and colonoids were cultured with lipopeptide Pam3CSK4, lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (Poly I:C) for 4 to 6 days. Surface area, budding and survival were assessed. Proliferation and numbers of lysozyme positive cells were quantified by flow cytometry. Gene expression was assessed by Nanostring and qRT-PCR. Results: Exposure to Pam3CSK4 and LPS had minimal impact on either enteroids or colonoids. In contrast, Poly I:C increased the surface area of enteroids, while colonoids demonstrated decreased budding. Survival was decreased by Poly I:C in enteroids but not in colonoids. Both enteroids and colonoids exhibited upregulated gene expression of chemokines, but these were increased in magnitude in enteroids. Decreases in gene expression associated with epithelial differentiation and lysozyme positive cells were more apparent in enteroids than in colonoids. Baseline gene expression between enteroids and colonoids differed markedly in levels of stem cell and inflammatory markers. The changes in morphology induced by Poly I:C were mediated by the toll-like receptor adaptor molecule 1 (Ticam1) in enteroids but not in colonoids. Conclusions: Poly I:C alters the molecular program of epithelial cells and shifts from absorption and digestion towards defense and inflammation. Diversity of responses to microbial patterns in enteroids and colonoids may underlie differences in susceptibility to infection along the intestinal tract. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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20. CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice.
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King, Jeffrey B., von Furstenberg, Richard J., Smith, Brian J., McNaughton, Kirk K., Galanko, Joseph A., and Henning, Susan J.
- Abstract
A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24+ fraction that contained the majority of Lgr5-EGFP+putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24+EpCAM+ fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase- like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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21. Mouse Background Strain Profoundly Influences Paneth Cell Function and Intestinal Microbial Composition.
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Gulati, Ajay S., Shanahanc, Michael T., Arthur, Janelle C., Grossniklaus, Emily, von Furstenberg, Richard J., Kreuk, Lieselotte, Henning, Susan J., Jobin, Christian, and Sartor, R. Balfour
- Subjects
CELL physiology ,BACTERIA ,HOMEOSTASIS ,LABORATORY mice ,PEPTIDES ,INTESTINES - Abstract
Background: Increasing evidence supports the central role of Paneth cells in maintaining intestinal host-microbial homeostasis. However, the direct impact of host genotype on Paneth cell function remains unclear. Here, we characterize key differences in Paneth cell function and intestinal microbial composition in two widely utilized, genetically distinct mouse strains (C57BL/6 and 129/SvEv). In doing so, we demonstrate critical influences of host genotype on Paneth cell activity and the enteric microbiota. Methodology and Principal Findings: Paneth cell numbers were determined by flow cytometry. Antimicrobial peptide (AMP) expression was evaluated using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), acid ureapolyacrylamide gel electrophoresis, and mass spectrometry. Effects of mouse background on microbial composition were assessed by reciprocal colonization of germ-free mice from both background strains, followed by compositional analysis of resultant gut bacterial communities using terminal restriction fragment length polymorphism analysis and 16 S qPCR. Our results revealed that 129/SvEv mice possessed fewer Paneth cells and a divergent AMP profile relative to C57BL/6 counterparts. Novel 129/SvEv á-defensin peptides were identified, including Defa2/18v, Defa11, Defa16, and Defa18. Host genotype profoundly affected the global profile of the intestinal microbiota, while both source and host factors were found to influence specific bacterial groups. Interestingly, ileal a-defensins from 129/SvEv mice displayed attenuated antimicrobial activity against pro-inflammatory E. coli strains, a bacterial species found to be expanded in these animals. Conclusions and Significance: This work establishes the important impact of host genotype on Paneth cell function and the composition of the intestinal microbiota. It further identifies specific AMP and microbial alterations in two commonly used inbred mouse strains that have varying susceptibilities to a variety of disorders, ranging from obesity to intestinal inflammation. This will be critical for future studies utilizing these murine backgrounds to study the effects of Paneth cells and the intestinal microbiota on host health and disease. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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22. Sorting mouse jejunal epithelial cells with CD24 yields a population with characteristics of intestinal stem cells.
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von Furstenberg, Richard J., Gulati, Ajay S., Baxi, Anand, Doherty, Jason M., Stappenbeck, Thaddeus S., Gracz, Adam D., Magness, Scott T., and Henning, Susan J.
- Subjects
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EPITHELIAL cells , *STEM cells , *ANTIGENS , *IMMUNOHISTOCHEMISTRY , *CELL adhesion , *REVERSE transcriptase polymerase chain reaction - Abstract
Intestinal stem cells (ISCs) have been studied for more than three decades; however, their isolation has remained a challenge. We hypothesized that, just as for stem cells of other tissues, one or more membrane markers would allow positive selection of ISCs by antibody-based sorting. To explore this hypothesis, microarray data of putative ISC fractions generated by side population sorting and laser capture microdissection were subjected to bioinformatic analysis to identify common membrane antigens. The microarray comparison suggested CD24 as a candidate surface marker, and immunohistochemistry showed expression of CD24 in epithelial cells of crypt bases. Flow cytometry of jejunal epithelial preparations revealed a CD24+ CD45- fraction comprising ~1% of the cells. Analysis with epithelial cell adhesion molecule and CD31 confirmed that the cell preparations were epithelial and without endothelial contamination. Cycling cells identified by prior injection with 5-ethynyl-2′-deoxyuridine were found predominantly in the CD24lo subfraction. Transcript analysis by real-time RT-PCR showed this subfraction to be enriched in the ISC markers leucine-rich-repeat-containing G-protein-coupled receptor 5 (40-fold) and Bmi1 (5-fold), but also enriched in lysozyme (10-fold). Flow cytometry with anti-lysozyme antibodies demonstrated that Paneth cells comprise ~30% of the CD24lo subfraction. Additional flow analyses with leucine-rich-repeat-containing G-protein-coupled receptor 5-enhanced green fluorescent protein (EGFP) epithelium demonstrated colocalization of EGFPhi and CD24lo. In contrast, CD24 cells were negative for the quiescent ISC marker doublecortin and CaM kinase-like-1. Culture of CD24lo cells in Matrigel generated organoid structures, which included all four epithelial lineages, thus giving functional evidence for the presence of ISCs. We conclude that the CD24lo fraction of jejunal epithelium is highly enriched with cycling ISCs. This isolation method should be useful to many investigators in the field to advance both the basic understanding of ISC biology and the therapeutic applications of ISCs. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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23. Tu1856 NOD2 Does Not Regulate Mouse Paneth Cell a-Defensin Expression
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Shanahan, Michael T., Grossniklaus, Emily, von Furstenberg, Richard J., Henning, Susan J., Sartor, Ryan B., and Gulati, Ajay S.
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- 2012
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24. CD24 as a Potential Marker to Directly Isolate LGR5+ Colonic Epithelial Cells
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King, Jeffrey B., von Furstenberg, Richard J., and Henning, Susan J.
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- 2011
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25. 787 Sorting With CD24 Yields a Population Markedly Enriched for Markers of Intestinal Stem Cells
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von Furstenberg, Richard J., Gulati, Ajay S., Stappenbeck, Thaddeus S., and Henning, Susan J.
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- 2010
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26. 523 Esophageal Submucosal Glands Contain Cells That Exhibit Progenitor Characteristics In Vitro.
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von Furstenberg, Richard J., Li, Joy, Henning, Susan J., and Garman, Katherine S.
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- 2016
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27. Tu1179 Previous Tonsillectomy Is Associated With Increased Risk of Esophageal Cancer.
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Garman, Katherine S., Boutte, Harold J., von Furstenberg, Richard J., Chiu, Shih-Ting, Lloyd, Benjamin, Zhang, Cecelia, Onaitis, Mark, Chow, Shein-Chung, and McCall, Shannon J.
- Published
- 2016
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28. Mo1253 Ductular and Proliferative Response of Esophageal Submucosal Glands in a Porcine Model of Esophageal Injury and Repair.
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Kruger, Leandi, Gonzalez, Liara M., von Furstenberg, Richard J., Henning, Susan J., Blikslager, Anthony, and Garman, Katherine S.
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- 2016
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29. Mo1836 Viral Products Elicit Different Phenotypes From Small Intestine and Colonic Crypt Cultures.
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Davies, Julie M., Santaolalla, Rebeca, von Furstenberg, Richard J., Henning, Susan J., and Abreu, Maria T.
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- 2015
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30. Intestinal Enteroendocrine Lineage Cells Possess Homeostatic and Injury-Inducible Stem Cell Activity.
- Author
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Yan KS, Gevaert O, Zheng GXY, Anchang B, Probert CS, Larkin KA, Davies PS, Cheng ZF, Kaddis JS, Han A, Roelf K, Calderon RI, Cynn E, Hu X, Mandleywala K, Wilhelmy J, Grimes SM, Corney DC, Boutet SC, Terry JM, Belgrader P, Ziraldo SB, Mikkelsen TS, Wang F, von Furstenberg RJ, Smith NR, Chandrakesan P, May R, Chrissy MAS, Jain R, Cartwright CA, Niland JC, Hong YK, Carrington J, Breault DT, Epstein J, Houchen CW, Lynch JP, Martin MG, Plevritis SK, Curtis C, Ji HP, Li L, Henning SJ, Wong MH, and Kuo CJ
- Subjects
- Animals, Antigens, Differentiation genetics, Enteroendocrine Cells pathology, Gene Expression Regulation, Intestinal Mucosa pathology, Jejunum pathology, Mice, Mice, Transgenic, Stem Cells pathology, Antigens, Differentiation metabolism, Enteroendocrine Cells metabolism, Intestinal Mucosa injuries, Intestinal Mucosa metabolism, Jejunum injuries, Jejunum metabolism, Stem Cells metabolism
- Abstract
Several cell populations have been reported to possess intestinal stem cell (ISC) activity during homeostasis and injury-induced regeneration. Here, we explored inter-relationships between putative mouse ISC populations by comparative RNA-sequencing (RNA-seq). The transcriptomes of multiple cycling ISC populations closely resembled Lgr5
+ ISCs, the most well-defined ISC pool, but Bmi1-GFP+ cells were distinct and enriched for enteroendocrine (EE) markers, including Prox1. Prox1-GFP+ cells exhibited sustained clonogenic growth in vitro, and lineage-tracing of Prox1+ cells revealed long-lived clones during homeostasis and after radiation-induced injury in vivo. Single-cell mRNA-seq revealed two subsets of Prox1-GFP+ cells, one of which resembled mature EE cells while the other displayed low-level EE gene expression but co-expressed tuft cell markers, Lgr5 and Ascl2, reminiscent of label-retaining secretory progenitors. Our data suggest that the EE lineage, including mature EE cells, comprises a reservoir of homeostatic and injury-inducible ISCs, extending our understanding of cellular plasticity and stemness., (Copyright © 2017 Elsevier Inc. All rights reserved.)- Published
- 2017
- Full Text
- View/download PDF
31. GI stem cells - new insights into roles in physiology and pathophysiology.
- Author
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Henning SJ and von Furstenberg RJ
- Subjects
- Animals, Cell Culture Techniques, Humans, Intestines transplantation, Intestinal Mucosa cytology, Stem Cells physiology
- Abstract
This overview gives a brief historical summary of key discoveries regarding stem cells of the small intestine. The current concept is that there are two pools of intestinal stem cells (ISCs): an actively cycling pool that is marked by Lgr5, is relatively homogeneous and is responsible for daily turnover of the epithelium; and a slowly cycling or quiescent pool that functions as reserve ISCs. The latter pool appears to be quite heterogeneous and may include partially differentiated epithelial lineages that can reacquire stem cell characteristics following injury to the intestine. Markers and methods of isolation for active and quiescent ISC populations are described as well as the numerous important advances that have been made in approaches to the in vitro culture of ISCs and crypts. Factors regulating ISC biology are briefly summarized and both known and unknown aspects of the ISC niche are discussed. Although most of our current knowledge regarding ISC physiology and pathophysiology has come from studies with mice, recent work with human tissue highlights the potential translational applications arising from this field of research. Many of these topics are further elaborated in the following articles., (© 2016 The Authors. The Journal of Physiology © 2016 The Physiological Society.)
- Published
- 2016
- Full Text
- View/download PDF
32. CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice.
- Author
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King JB, von Furstenberg RJ, Smith BJ, McNaughton KK, Galanko JA, and Henning SJ
- Subjects
- Animals, Antigens, Neoplasm metabolism, Biomarkers metabolism, Cell Adhesion Molecules metabolism, Cell Proliferation, Colon cytology, Epithelial Cell Adhesion Molecule, Gene Expression Regulation, Green Fluorescent Proteins biosynthesis, Green Fluorescent Proteins genetics, Immunohistochemistry, Intestinal Mucosa cytology, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Confocal, RNA, Messenger metabolism, Receptors, G-Protein-Coupled genetics, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic, CD24 Antigen metabolism, Cell Separation methods, Colon immunology, Epithelial Cells immunology, Flow Cytometry, Intestinal Mucosa immunology, Receptors, G-Protein-Coupled metabolism, Stem Cells immunology
- Abstract
A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24(+) fraction that contained the majority of Lgr5-EGFP(+) putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24(+)EpCAM(+) fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase-like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies.
- Published
- 2012
- Full Text
- View/download PDF
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