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  1. Article ; Online: Has a physiologic function for NHE2 finally been identified?

    Donowitz, Mark

    Acta physiologica (Oxford, England)

    2022  Volume 234, Issue 3, Page(s) e13792

    MeSH term(s) Hydrogen ; Intestinal Mucosa ; RNA, Messenger ; Sodium
    Chemical Substances RNA, Messenger ; Hydrogen (7YNJ3PO35Z) ; Sodium (9NEZ333N27)
    Language English
    Publishing date 2022-02-11
    Publishing country England
    Document type Editorial ; Comment
    ZDB-ID 2218636-0
    ISSN 1748-1716 ; 1748-1708
    ISSN (online) 1748-1716
    ISSN 1748-1708
    DOI 10.1111/apha.13792
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  2. Article ; Online: Presentation of the AGA Distinguished Achievement Award in Basic Science to Kim E. Barrett, PhD, AGAF.

    Donowitz, Mark / Keely, Stephen

    Gastroenterology

    2021  Volume 161, Issue 1, Page(s) 336–338

    MeSH term(s) Awards and Prizes ; Biomedical Research/history ; Gastroenterology/history ; History, 20th Century ; History, 21st Century ; Humans
    Language English
    Publishing date 2021-05-21
    Publishing country United States
    Document type Biography ; Editorial ; Historical Article ; Portrait
    ZDB-ID 80112-4
    ISSN 1528-0012 ; 0016-5085
    ISSN (online) 1528-0012
    ISSN 0016-5085
    DOI 10.1053/j.gastro.2021.05.037
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  3. Article: Practical Advice From a Basic and Translational Researcher at a Major Medical School.

    Donowitz, Mark

    Cellular and molecular gastroenterology and hepatology

    2016  Volume 2, Issue 6, Page(s) 707–709

    Language English
    Publishing date 2016-11
    Publishing country United States
    Document type Editorial
    ISSN 2352-345X
    ISSN 2352-345X
    DOI 10.1016/j.jcmgh.2016.08.002
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  4. Article ; Online: The Air-Liquid Interface Reorganizes Membrane Lipids and Enhances the Recruitment of Slc26a3 to Lipid-Rich Domains in Human Colonoid Monolayers.

    Tse, C Ming / Zhang, Zixin / Lin, Ruxian / Sarker, Rafiquel / Donowitz, Mark / Singh, Varsha

    International journal of molecular sciences

    2023  Volume 24, Issue 9

    Abstract: Cholesterol-rich membrane domains, also called lipid rafts (LRs), are specialized membrane domains that provide a platform for intracellular signal transduction. Membrane proteins often cluster in LRs that further aggregate into larger platform-like ... ...

    Abstract Cholesterol-rich membrane domains, also called lipid rafts (LRs), are specialized membrane domains that provide a platform for intracellular signal transduction. Membrane proteins often cluster in LRs that further aggregate into larger platform-like structures that are enriched in ceramides and are called ceramide-rich platforms (CRPs). The role of CRPs in the regulation of intestinal epithelial functions remains unknown. Down-regulated in adenoma (DRA) is an intestinal Cl
    MeSH term(s) Humans ; Caco-2 Cells ; Chloride-Bicarbonate Antiporters/metabolism ; Membrane Lipids ; Antiporters/metabolism ; Cell Differentiation ; Sulfate Transporters/metabolism
    Chemical Substances Chloride-Bicarbonate Antiporters ; Membrane Lipids ; Antiporters ; SLC26A3 protein, human ; Sulfate Transporters
    Language English
    Publishing date 2023-05-05
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms24098273
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  5. Article ; Online: SLC26A3 (DRA) is stimulated in a synergistic, intracellular Ca

    Sarker, Rafiquel / Lin, Ruxian / Singh, Varsha / Donowitz, Mark / Tse, Chung-Ming

    American journal of physiology. Cell physiology

    2023  Volume 324, Issue 6, Page(s) C1263–C1273

    Abstract: In polarized intestinal epithelial cells, downregulated in adenoma (DRA) is an apical ... ...

    Abstract In polarized intestinal epithelial cells, downregulated in adenoma (DRA) is an apical Cl
    MeSH term(s) Humans ; Caco-2 Cells ; Sodium Chloride ; Epithelial Cells/metabolism ; Anions/metabolism ; Sodium-Hydrogen Exchanger 3/metabolism ; Diarrhea/metabolism ; Adenosine Triphosphate/pharmacology ; Adenosine Triphosphate/metabolism ; Sulfate Transporters/genetics ; Sulfate Transporters/metabolism ; Chloride-Bicarbonate Antiporters/genetics ; Chloride-Bicarbonate Antiporters/metabolism
    Chemical Substances 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester (139890-68-9) ; Sodium Chloride (451W47IQ8X) ; Anions ; Sodium-Hydrogen Exchanger 3 ; 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (K22DDW77C0) ; Adenosine Triphosphate (8L70Q75FXE) ; SLC26A3 protein, human ; Sulfate Transporters ; Chloride-Bicarbonate Antiporters
    Language English
    Publishing date 2023-05-08
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 392098-7
    ISSN 1522-1563 ; 0363-6143
    ISSN (online) 1522-1563
    ISSN 0363-6143
    DOI 10.1152/ajpcell.00523.2022
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  6. Article ; Online: Intestinal enteroids/organoids: A novel platform for drug discovery in inflammatory bowel diseases.

    Yoo, Jun-Hwan / Donowitz, Mark

    World journal of gastroenterology

    2019  Volume 25, Issue 30, Page(s) 4125–4147

    Abstract: The introduction of biologics such as anti-tumor necrosis factor (TNF) monoclonal antibodies followed by anti-integrins has dramatically changed the therapeutic paradigm of inflammatory bowel diseases (IBD). Furthermore, a newly developed anti-p40 ... ...

    Abstract The introduction of biologics such as anti-tumor necrosis factor (TNF) monoclonal antibodies followed by anti-integrins has dramatically changed the therapeutic paradigm of inflammatory bowel diseases (IBD). Furthermore, a newly developed anti-p40 subunit of interleukin (IL)-12 and IL-23 (ustekinumab) has been recently approved in the United States for patients with moderate to severe Crohn's disease who have failed treatment with anti-TNFs. However, these immunosuppressive therapeutics which focus on anti-inflammatory mechanisms or immune cells still fail to achieve long-term remission in a significant percentage of patients. This strongly underlines the need to identify novel treatment targets beyond immune suppression to treat IBD. Recent studies have revealed the critical role of intestinal epithelial cells (IECs) in the pathogenesis of IBD. Physical, biochemical and immunologic driven barrier dysfunctions of epithelial cells contribute to the development of IBD. In addition, the recent establishment of adult stem cell-derived intestinal enteroid/organoid culture technology has allowed an exciting opportunity to study human IECs comprising all normal epithelial cells. This long-term epithelial culture model can be generated from endoscopic biopsies or surgical resections and recapitulates the tissue of origin, representing a promising platform for novel drug discovery in IBD. This review describes the advantages of intestinal enteroids/organoids as a research tool for intestinal diseases, introduces studies with these models in IBD, and gives a description of the current status of therapeutic approaches in IBD. Finally, we provide an overview of the current endeavors to identify a novel drug target for IBD therapy based on studies with human enteroids/organoids and describe the challenges in using enteroids/organoids as an IBD model.
    MeSH term(s) Biological Products/pharmacology ; Biological Products/therapeutic use ; Biopsy ; Drug Discovery/methods ; Drug Evaluation, Preclinical/methods ; Epithelial Cells ; Humans ; Immunosuppressive Agents/pharmacology ; Immunosuppressive Agents/therapeutic use ; Induced Pluripotent Stem Cells ; Inflammatory Bowel Diseases/drug therapy ; Inflammatory Bowel Diseases/immunology ; Inflammatory Bowel Diseases/pathology ; Intestinal Mucosa/cytology ; Intestinal Mucosa/drug effects ; Intestinal Mucosa/immunology ; Organoids/drug effects ; Organoids/immunology ; Primary Cell Culture/methods
    Chemical Substances Biological Products ; Immunosuppressive Agents
    Language English
    Publishing date 2019-07-24
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 2185929-2
    ISSN 2219-2840 ; 1007-9327
    ISSN (online) 2219-2840
    ISSN 1007-9327
    DOI 10.3748/wjg.v25.i30.4125
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  7. Article ; Online: Patients' perspectives on the derivation and use of organoids.

    Bollinger, Juli / May, Elizabeth / Mathews, Debra / Donowitz, Mark / Sugarman, Jeremy

    Stem cell reports

    2021  Volume 16, Issue 8, Page(s) 1874–1883

    Abstract: Organoid research is enhancing understanding of human development and diseases as well as aiding in medication development and selection, raising hopes for even more future therapeutic options. Nevertheless, this work raises important ethical issues and ... ...

    Abstract Organoid research is enhancing understanding of human development and diseases as well as aiding in medication development and selection, raising hopes for even more future therapeutic options. Nevertheless, this work raises important ethical issues and there is a paucity of data regarding patients' perspectives on them. We report on 60 interviews with adult patients or parents of pediatric patients from diverse disease populations who receive medical care at a major academic research institution in the United States. Interviewees expressed broad support for organoid development and use. However, patients viewed brain organoids, and sometimes gonadal organoids, as morally distinct; and some organoid research poses moral concerns. Nonetheless, patients generally understood the potential value of such research and approved of it, provided it was aimed at good intent and conducted with ethical oversight and a robust consent process. These data should help inform conceptual and policy deliberations about appropriate organoid use.
    MeSH term(s) Adult ; Aged ; Biomedical Research/ethics ; Biomedical Research/methods ; Brain/cytology ; Brain/metabolism ; Female ; Gonads/cytology ; Gonads/metabolism ; Humans ; Interviews as Topic ; Male ; Middle Aged ; Organoids/cytology ; Organoids/metabolism ; Patients/psychology ; Surveys and Questionnaires ; Young Adult
    Language English
    Publishing date 2021-07-29
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2720528-9
    ISSN 2213-6711 ; 2213-6711
    ISSN (online) 2213-6711
    ISSN 2213-6711
    DOI 10.1016/j.stemcr.2021.07.004
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  8. Article: Identification of Intestinal NaCl Absorptive-Anion Secretory Cells: Potential Functional Significance.

    Donowitz, Mark / Sarker, Rafiquel / Lin, Ruxian / McNamara, George / Tse, Chung Ming / Singh, Varsha

    Frontiers in physiology

    2022  Volume 13, Page(s) 892112

    Abstract: Use of human enteroids studied in the undifferentiated and differentiated state that mimic the intestinal crypt and villus, respectively, has allowed studies of multiple enterocyte populations, including a large population of enterocytes that are ... ...

    Abstract Use of human enteroids studied in the undifferentiated and differentiated state that mimic the intestinal crypt and villus, respectively, has allowed studies of multiple enterocyte populations, including a large population of enterocytes that are transitioning from the crypt to the villus. This population expresses NHE3, DRA, and CFTR, representing a combination of Na absorptive and anion secretory functions. In this cell population, these three transporters physically interact, which affects their baseline and regulated activities. A study of this cell population and differentiated Caco-2 cells transduced with NHE3 and endogenously expressing DRA and CFTR has allowed an understanding of previous studies in which cAMP seemed to stimulate and inhibit DRA at the same time. Understanding the contributions of these cells to overall intestinal transport function as part of the fasting and post-prandial state and their contribution to the pathophysiology of diarrheal diseases and some conditions with constipation will allow new approaches to drug development.
    Language English
    Publishing date 2022-07-19
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2564217-0
    ISSN 1664-042X
    ISSN 1664-042X
    DOI 10.3389/fphys.2022.892112
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  9. Article ; Online: mOrange2, a Genetically Encoded, pH Sensitive Fluorescent Protein, is an Alternative to BCECF-AM to Measure Intracellular pH to Determine NHE3 and DRA Activity.

    Sarker, Rafiquel / Tse, Chung Ming / Lin, Ruxian / McNamara, George / Singh, Varsha / Donowitz, Mark

    Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

    2022  Volume 56, Issue 1, Page(s) 39–49

    Abstract: Background/aims: NHE3 (Na: Methods: The use of a genetically modified fluorescent protein, mOrange2 was explored as an intracellular pH sensor protein to measure exchange activity of NHE3 and DRA. The model used was FRT cells stably expressing NHE3 ... ...

    Abstract Background/aims: NHE3 (Na
    Methods: The use of a genetically modified fluorescent protein, mOrange2 was explored as an intracellular pH sensor protein to measure exchange activity of NHE3 and DRA. The model used was FRT cells stably expressing NHE3 or DRA with intracellular pH measured by changes of mOrange2 fluorescence intensity. Intracellular pH was monitored using a) Isolated single clones of FRT/mOrange2/HA-NHE3 cells studied in a confocal microscope with time-lapse live cell imaging under basal conditions and when NHE3 was inhibited by exposure to forskolin and stimulated by dexamethasone, b) coverslip grown FRT/mOrange2 cells expressing NHE3 or DRA using a computerized fluorometer with a perfused cuvette with standardization of the mOrange2 absorption and emission signal using K
    Results: A similar rate of intracellular alkalization by Na
    Conclusion: This study demonstrates that mOrange2 protein can be an effective alternate to BCECF-AM in measuring intracellular pH (preferred setting Ex520nm, Em 563nm) as affected by NHE3 and DRA activity, with the advantage, compared to AM ester dyes, that genetic expression can provide uniform expression of the pH sensor.
    MeSH term(s) Animals ; Antiporters/genetics ; Antiporters/metabolism ; Fluoresceins/pharmacology ; Hydrogen-Ion Concentration ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism ; Rats ; Rats, Inbred F344 ; Sodium-Hydrogen Exchanger 3/genetics ; Sodium-Hydrogen Exchanger 3/metabolism ; Sulfate Transporters/genetics ; Sulfate Transporters/metabolism
    Chemical Substances Antiporters ; Fluoresceins ; Luminescent Proteins ; Slc26a3 protein, rat ; Slc9a3 protein, rat ; Sodium-Hydrogen Exchanger 3 ; Sulfate Transporters ; fluorescent protein 583 ; 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (85138-49-4)
    Language English
    Publishing date 2022-01-25
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 1067572-3
    ISSN 1421-9778 ; 1015-8987
    ISSN (online) 1421-9778
    ISSN 1015-8987
    DOI 10.33594/000000493
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  10. Article ; Online: Calcium-sensing receptor activator cinacalcet for treatment of cyclic nucleotide-mediated secretory diarrheas.

    Chu, Tifany / Yottasan, Pattareeya / Goncalves, Livia de Souza / Oak, Apurva A / Lin, Ruxian / Tse, Ming / Donowitz, Mark / Cil, Onur

    Translational research : the journal of laboratory and clinical medicine

    2023  Volume 263, Page(s) 45–52

    Abstract: Cyclic nucleotide elevation in intestinal epithelial cells is the key pathology causing intestinal fluid loss in secretory diarrheas such as cholera. Current secretory diarrhea treatment is primarily supportive, and oral rehydration solution is the ... ...

    Abstract Cyclic nucleotide elevation in intestinal epithelial cells is the key pathology causing intestinal fluid loss in secretory diarrheas such as cholera. Current secretory diarrhea treatment is primarily supportive, and oral rehydration solution is the mainstay of cholera treatment. There is an unmet need for safe, simple and effective diarrhea treatments. By promoting cAMP hydrolysis, extracellular calcium-sensing receptor (CaSR) is a regulator of intestinal fluid transport. We studied the antidiarrheal mechanisms of FDA-approved CaSR activator cinacalcet and tested its efficacy in clinically relevant human cell, mouse and intestinal organoid models of secretory diarrhea. By using selective inhibitors, we found that cAMP agonists-induced secretory short-circuit currents (I
    MeSH term(s) Mice ; Humans ; Animals ; Antidiarrheals/metabolism ; Antidiarrheals/pharmacology ; Antidiarrheals/therapeutic use ; Cholera/drug therapy ; Cholera/metabolism ; Cholera/pathology ; Cholera Toxin/metabolism ; Cholera Toxin/pharmacology ; Cholera Toxin/therapeutic use ; Cinacalcet/pharmacology ; Cinacalcet/therapeutic use ; Cinacalcet/metabolism ; Receptors, Calcium-Sensing/metabolism ; Receptors, Calcium-Sensing/therapeutic use ; Nucleotides, Cyclic/metabolism ; Nucleotides, Cyclic/pharmacology ; Nucleotides, Cyclic/therapeutic use ; Colforsin/metabolism ; Colforsin/pharmacology ; Colforsin/therapeutic use ; Diarrhea/drug therapy ; Diarrhea/metabolism ; Intestinal Mucosa/metabolism ; Cystic Fibrosis Transmembrane Conductance Regulator/metabolism ; Cystic Fibrosis Transmembrane Conductance Regulator/therapeutic use ; Mice, Knockout
    Chemical Substances Antidiarrheals ; Cholera Toxin (9012-63-9) ; Cinacalcet (UAZ6V7728S) ; Receptors, Calcium-Sensing ; Nucleotides, Cyclic ; Colforsin (1F7A44V6OU) ; Cystic Fibrosis Transmembrane Conductance Regulator (126880-72-6)
    Language English
    Publishing date 2023-09-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2246684-8
    ISSN 1878-1810 ; 1532-6543 ; 1931-5244
    ISSN (online) 1878-1810 ; 1532-6543
    ISSN 1931-5244
    DOI 10.1016/j.trsl.2023.09.001
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