{"doi":"10.1016/j.jcmgh.2020.03.005","title":"Protein Kinase D1 (PKD1) Signaling Induces Growth-Promoting Effects in Murine Enteroids","abstract":"The mammalian intestine, covered by a single layer of epithelial cells, renews every 4–6 days throughout adult life. This high rate of turnover plays an essential role in the organization, maintenance, function, and restoration of intestinal tissue integrity. The ultimate source of cells are multipotent intestinal stem cells (ISCs) that reside in the lower regions of intestinal crypts.1Koo B.-K. et al.Gastroenterology. 2014; 147: 289-302Abstract Full Text Full Text PDF PubMed Scopus (111) Google Scholar Despite major advances in the identification of rapidly dividing ISCs and their regulation through growth factors and developmental cues from the microenvironment,1Koo B.-K. et al.Gastroenterology. 2014; 147: 289-302Abstract Full Text Full Text PDF PubMed Scopus (111) Google Scholar, 2Ootani A. et al.Nat Med. 2009; 15: 701-706Crossref PubMed Scopus (618) Google Scholar, 3Sato T. et al.Nature. 2009; 459: 262-265Crossref PubMed Scopus (4154) Google Scholar, 4Sato T. et al.Nature. 2011; 469: 415-418Crossref PubMed Scopus (1733) Google Scholar the intracellular signal transduction mechanisms remain incompletely understood. Protein kinase D1 (PKD1), an evolutionarily conserved protein kinase family, has emerged as a key node in cellular signaling.5Rozengurt E. et al.J Biol Chem. 2005; 280: 13205-13208Crossref PubMed Scopus (366) Google Scholar,6Rozengurt E. Physiology. 2011; 26: 23-33Crossref PubMed Scopus (188) Google Scholar By using transgenic mice that overexpress PKD1 in the intestine (PKD1-Tg), we showed that PKD1 induces crypt cell hyperproliferation and leads to a change in tissue architecture, manifested by an increase in the size and total number of epithelial cells in intestinal crypts.7Sinnett-Smith J. et al.J Biol Chem. 2011; 286: 511-520Crossref PubMed Scopus (16) Google Scholar,8Wang J. et al.Am J Physiol Cell Physiol. 2016; 310: C542-C557Crossref PubMed Scopus (9) Google Scholar These findings prompted us to hypothesize that PKD1 controls the number and function of ISCs, a proposition that remained untested. Here, we determined the impact of PKD1 activity on the function of ISCs by assessing the capacity of isolated intestinal crypts to form 3-dimensional enteroids.2Ootani A. et al.Nat Med. 2009; 15: 701-706Crossref PubMed Scopus (618) Google Scholar,3Sato T. et al.Nature. 2009; 459: 262-265Crossref PubMed Scopus (4154) Google Scholar We isolated crypts from PKD1-Tg and non-Tg littermates and plated them in Matrigel (Corning, Tewksbury, MA) containing R-spondin, noggin, and epidermal growth factor.3Sato T. et al.Nature. 2009; 459: 262-265Crossref PubMed Scopus (4154) Google Scholar Initially, we verified that PKD1 expression and activity were markedly more pronounced in lysates of PKD1-Tg enteroids, as shown by Western blotting with antibodies that detect total PKD1 and catalytically active PKD15Rozengurt E. et al.J Biol Chem. 2005; 280: 13205-13208Crossref PubMed Scopus (366) Google Scholar,6Rozengurt E. Physiology. 2011; 26: 23-33Crossref PubMed Scopus (188) Google Scholar phosphorylated at the activation loop Ser744/748 or at the C-terminal autophosphorylation site Ser916 (Figure 1A). Enteroids generated from PKD1-Tg mice show a significant increase in area compared with enteroids generated from crypts isolated from non-Tg mice (Figure 1B and Supplementary Figure 1). After 7 days, the area of enteroids formed from PKD1-Tg mice was 58 ± 2.3 × 103 μm2 compared with 42 ± 3.5 × 103 μm2 in enteroids from non-Tg mice (Figure 1C). Similarly, the number of crypt-like buds, which contain the stem cell compartment,3Sato T. et al.Nature. 2009; 459: 262-265Crossref PubMed Scopus (4154) Google Scholar was 8.1 ± 0.49 in enteroids from PKD1-Tg and 5.5 ± 0.31 in those from non-Tg mice (Figure 1D). Importantly, we obtained similar results with enteroids passaged 4 times in culture (Figure 1B–D), or with enteroids prepared from primary crypts from PKD1-Tg and non-Tg mice (Figure 1E–G). Given that the impact of PKD1 overexpression o","journal":"Cellular and Molecular Gastroenterology and Hepatology","year":2020,"id":112317,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9461,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":531727,"name":"James Sinnett‐Smith","orcid":"0000-0003-0074-7270","position":1,"is_corresponding":false},{"id":531728,"name":"Michelle K. Tenggara","orcid":"0000-0001-5582-7775","position":2,"is_corresponding":false},{"id":531729,"name":"M. Martín","orcid":"0000-0002-6326-8405","position":3,"is_corresponding":false},{"id":405221,"name":"Enrique Rozengurt","orcid":"0000-0002-7838-9170","position":4,"is_corresponding":false},{"id":531726,"name":"Yuki Shimizu","orcid":"0000-0003-0938-3135","position":0,"is_corresponding":true}],"reference_count":13,"raw_metadata":null,"created_at":"2026-07-18T23:13:13.874980Z","pmid":"32234448","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}