Development of a reliable automated screening system to identify small molecules and biologics that promote human β-cell regeneration
Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell proliferation in response to such compounds, we developed a medium-throughput in vitro method of quantifying adult human β-cell proliferation ma...
Saved in:
| Published in: | American journal of physiology: endocrinology and metabolism Vol. 311; no. 5; p. E859 |
|---|---|
| Main Authors: | , , , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
United States
01.11.2016
|
| Subjects: | |
| ISSN: | 1522-1555, 1522-1555 |
| Online Access: | Get more information |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Abstract | Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell proliferation in response to such compounds, we developed a medium-throughput in vitro method of quantifying adult human β-cell proliferation markers. This method is based on high-content imaging of dispersed islet cells seeded in 384-well plates and automated cell counting that identifies fluorescently labeled β-cells with high specificity using both nuclear and cytoplasmic markers. β-Cells from each donor were assessed for their function and ability to enter the cell cycle by cotransduction with adenoviruses encoding cell cycle regulators cdk6 and cyclin D3. Using this approach, we tested 12 previously identified mitogens, including neurotransmitters, hormones, growth factors, and molecules, involved in adenosine and Tgf-1β signaling. Each compound was tested in a wide concentration range either in the presence of basal (5 mM) or high (11 mM) glucose. Treatment with the control compound harmine, a Dyrk1a inhibitor, led to a significant increase in Ki-67
β-cells, whereas treatment with other compounds had limited to no effect on human β-cell proliferation. This new scalable approach reduces the time and effort required for sensitive and specific evaluation of human β-cell proliferation, thus allowing for increased testing of candidate human β-cell mitogens. |
|---|---|
| AbstractList | Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell proliferation in response to such compounds, we developed a medium-throughput in vitro method of quantifying adult human β-cell proliferation markers. This method is based on high-content imaging of dispersed islet cells seeded in 384-well plates and automated cell counting that identifies fluorescently labeled β-cells with high specificity using both nuclear and cytoplasmic markers. β-Cells from each donor were assessed for their function and ability to enter the cell cycle by cotransduction with adenoviruses encoding cell cycle regulators cdk6 and cyclin D3. Using this approach, we tested 12 previously identified mitogens, including neurotransmitters, hormones, growth factors, and molecules, involved in adenosine and Tgf-1β signaling. Each compound was tested in a wide concentration range either in the presence of basal (5 mM) or high (11 mM) glucose. Treatment with the control compound harmine, a Dyrk1a inhibitor, led to a significant increase in Ki-67+ β-cells, whereas treatment with other compounds had limited to no effect on human β-cell proliferation. This new scalable approach reduces the time and effort required for sensitive and specific evaluation of human β-cell proliferation, thus allowing for increased testing of candidate human β-cell mitogens.Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell proliferation in response to such compounds, we developed a medium-throughput in vitro method of quantifying adult human β-cell proliferation markers. This method is based on high-content imaging of dispersed islet cells seeded in 384-well plates and automated cell counting that identifies fluorescently labeled β-cells with high specificity using both nuclear and cytoplasmic markers. β-Cells from each donor were assessed for their function and ability to enter the cell cycle by cotransduction with adenoviruses encoding cell cycle regulators cdk6 and cyclin D3. Using this approach, we tested 12 previously identified mitogens, including neurotransmitters, hormones, growth factors, and molecules, involved in adenosine and Tgf-1β signaling. Each compound was tested in a wide concentration range either in the presence of basal (5 mM) or high (11 mM) glucose. Treatment with the control compound harmine, a Dyrk1a inhibitor, led to a significant increase in Ki-67+ β-cells, whereas treatment with other compounds had limited to no effect on human β-cell proliferation. This new scalable approach reduces the time and effort required for sensitive and specific evaluation of human β-cell proliferation, thus allowing for increased testing of candidate human β-cell mitogens. Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell proliferation in response to such compounds, we developed a medium-throughput in vitro method of quantifying adult human β-cell proliferation markers. This method is based on high-content imaging of dispersed islet cells seeded in 384-well plates and automated cell counting that identifies fluorescently labeled β-cells with high specificity using both nuclear and cytoplasmic markers. β-Cells from each donor were assessed for their function and ability to enter the cell cycle by cotransduction with adenoviruses encoding cell cycle regulators cdk6 and cyclin D3. Using this approach, we tested 12 previously identified mitogens, including neurotransmitters, hormones, growth factors, and molecules, involved in adenosine and Tgf-1β signaling. Each compound was tested in a wide concentration range either in the presence of basal (5 mM) or high (11 mM) glucose. Treatment with the control compound harmine, a Dyrk1a inhibitor, led to a significant increase in Ki-67 β-cells, whereas treatment with other compounds had limited to no effect on human β-cell proliferation. This new scalable approach reduces the time and effort required for sensitive and specific evaluation of human β-cell proliferation, thus allowing for increased testing of candidate human β-cell mitogens. |
| Author | Wang, Peng Aramandla, Radhika Brissova, Marcela Powers, Alvin C Aamodt, Kristie I Brown, Judy J Fiaschi-Taesch, Nathalie Stewart, Andrew F |
| Author_xml | – sequence: 1 givenname: Kristie I surname: Aamodt fullname: Aamodt, Kristie I organization: Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, Tennessee – sequence: 2 givenname: Radhika surname: Aramandla fullname: Aramandla, Radhika organization: Division of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee – sequence: 3 givenname: Judy J surname: Brown fullname: Brown, Judy J organization: Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee – sequence: 4 givenname: Nathalie surname: Fiaschi-Taesch fullname: Fiaschi-Taesch, Nathalie organization: Division of Endocrinology, Diabetes, and Bone Disease, Department of Medicine, Mount Sinai Medical Center, New York, New York; and – sequence: 5 givenname: Peng surname: Wang fullname: Wang, Peng organization: Division of Endocrinology, Diabetes, and Bone Disease, Department of Medicine, Mount Sinai Medical Center, New York, New York; and – sequence: 6 givenname: Andrew F surname: Stewart fullname: Stewart, Andrew F organization: Division of Endocrinology, Diabetes, and Bone Disease, Department of Medicine, Mount Sinai Medical Center, New York, New York; and – sequence: 7 givenname: Marcela surname: Brissova fullname: Brissova, Marcela email: marcela.brissova@vanderbilt.edu organization: Division of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee; marcela.brissova@vanderbilt.edu – sequence: 8 givenname: Alvin C surname: Powers fullname: Powers, Alvin C organization: Veterans Affairs Tennessee Valley Healthcare System, Nashville, Tennessee |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/27624103$$D View this record in MEDLINE/PubMed |
| BookMark | eNpNkE1OwzAQhS1URH_gAiyQl2xSbNdunCUqv1IlNrCObGfSuvJPiR2kXoADcRDORBAgsZonzftmnt4UjUIMgNA5JXNKBbtSuz2EJs4JEVTMGaHiCE2GBSuoEGL0T4_RNKUdIaQUnJ2gMSuXjFOymKD3G3gDF_ceQsaxxQp34KzSDrDqc_QqQ4OT6QCCDRucDimDxzli2wyEbQ84eeUc9tGB6R0krEKDtY0ubqxJOG9Vxvsu-pgBb3uvAv78KAwMSAcbCNCpbGM4RcetcgnOfucMvdzdPq8eivXT_ePqel0YTlguVEkbWUkiW0FaKbhsSzBLoyoClZbAtKSMSV62hMuF1qXUrWBqaQyRFS85sBm6_Lk7RHrtIeXa2_SdRgWIfaqp5MMjKRZisF78Wnvtoan3nfWqO9R_3bEvyRJ3YQ |
| CitedBy_id | crossref_primary_10_1016_j_cmet_2018_10_007 crossref_primary_10_1016_j_cophys_2019_10_019 crossref_primary_10_1210_endocr_bqac193 crossref_primary_10_1016_j_biopha_2022_112957 crossref_primary_10_1038_s41598_018_33875_0 crossref_primary_10_3390_cells12060849 crossref_primary_10_3390_molecules25081983 crossref_primary_10_1155_2019_5783545 crossref_primary_10_3389_fcell_2021_734597 crossref_primary_10_3390_ijms22073306 crossref_primary_10_3389_fendo_2025_1612576 crossref_primary_10_1016_j_cmet_2017_05_011 crossref_primary_10_3390_ijms21218244 crossref_primary_10_1016_j_cmet_2019_12_009 crossref_primary_10_1002_jcp_28214 crossref_primary_10_1016_j_cmet_2018_12_005 crossref_primary_10_1080_19382014_2024_2334044 crossref_primary_10_3389_fendo_2022_854094 crossref_primary_10_1016_j_jbc_2025_110645 crossref_primary_10_1016_j_transproceed_2018_12_028 crossref_primary_10_1172_JCI144833 crossref_primary_10_3389_fendo_2021_636182 crossref_primary_10_1007_s00125_018_4639_6 crossref_primary_10_1016_j_cbpa_2022_102150 crossref_primary_10_1038_s41589_024_01822_y crossref_primary_10_1007_s12020_021_02941_5 crossref_primary_10_1038_s41598_021_90643_3 crossref_primary_10_1111_dom_13031 |
| ContentType | Journal Article |
| DBID | CGR CUY CVF ECM EIF NPM 7X8 |
| DOI | 10.1152/ajpendo.00515.2015 |
| DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed MEDLINE - Academic |
| DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) MEDLINE - Academic |
| DatabaseTitleList | MEDLINE - Academic MEDLINE |
| Database_xml | – sequence: 1 dbid: NPM name: PubMed url: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: 7X8 name: MEDLINE - Academic url: https://search.proquest.com/medline sourceTypes: Aggregation Database |
| DeliveryMethod | no_fulltext_linktorsrc |
| Discipline | Medicine Anatomy & Physiology |
| EISSN | 1522-1555 |
| ExternalDocumentID | 27624103 |
| Genre | Journal Article |
| GrantInformation_xml | – fundername: NIDDK NIH HHS grantid: R01 DK105015 – fundername: NIDDK NIH HHS grantid: UC4 DK104211 – fundername: NIDDK NIH HHS grantid: P30 DK020541 – fundername: NIDDK NIH HHS grantid: P30 DK020593 – fundername: NIGMS NIH HHS grantid: T32 GM007347 – fundername: NIH HHS grantid: S10 OD021630 – fundername: NIDDK NIH HHS grantid: F30 DK097921 – fundername: NIDDK NIH HHS grantid: R24 DK106755 |
| GroupedDBID | --- 23M 2WC 39C 4.4 53G 5GY 5VS 6J9 AAFWJ ABJNI ACPRK ADBBV AENEX AFRAH ALMA_UNASSIGNED_HOLDINGS BAWUL BKKCC BKOMP BTFSW CGR CUY CVF DIK E3Z EBS ECM EIF EJD EMOBN F5P GX1 H13 ITBOX KQ8 NPM OK1 P2P P6G PQQKQ RAP RHF RHI RPL RPRKH TR2 W8F WH7 WOQ XSW YSK 7X8 |
| ID | FETCH-LOGICAL-c402t-a71d89808f50f8548f7ec6ca90e9b8e2b8122847f0483bb78bf52a6cc089474e2 |
| IEDL.DBID | 7X8 |
| ISICitedReferencesCount | 32 |
| ISICitedReferencesURI | http://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=Summon&SrcAuth=ProQuest&DestLinkType=CitingArticles&DestApp=WOS_CPL&KeyUT=000390110800007&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| ISSN | 1522-1555 |
| IngestDate | Wed Oct 15 13:45:23 EDT 2025 Thu Jan 02 22:58:57 EST 2025 |
| IsDoiOpenAccess | false |
| IsOpenAccess | true |
| IsPeerReviewed | true |
| IsScholarly | true |
| Issue | 5 |
| Keywords | β-cell proliferation human islet |
| Language | English |
| LinkModel | DirectLink |
| MergedId | FETCHMERGED-LOGICAL-c402t-a71d89808f50f8548f7ec6ca90e9b8e2b8122847f0483bb78bf52a6cc089474e2 |
| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| OpenAccessLink | https://www.physiology.org/doi/pdf/10.1152/ajpendo.00515.2015 |
| PMID | 27624103 |
| PQID | 1844028535 |
| PQPubID | 23479 |
| ParticipantIDs | proquest_miscellaneous_1844028535 pubmed_primary_27624103 |
| PublicationCentury | 2000 |
| PublicationDate | 2016-11-01 |
| PublicationDateYYYYMMDD | 2016-11-01 |
| PublicationDate_xml | – month: 11 year: 2016 text: 2016-11-01 day: 01 |
| PublicationDecade | 2010 |
| PublicationPlace | United States |
| PublicationPlace_xml | – name: United States |
| PublicationTitle | American journal of physiology: endocrinology and metabolism |
| PublicationTitleAlternate | Am J Physiol Endocrinol Metab |
| PublicationYear | 2016 |
| SSID | ssj0007542 |
| Score | 2.368912 |
| Snippet | Numerous compounds stimulate rodent β-cell proliferation; however, translating these findings to human β-cells remains a challenge. To examine human β-cell... |
| SourceID | proquest pubmed |
| SourceType | Aggregation Database Index Database |
| StartPage | E859 |
| SubjectTerms | Activins - pharmacology Adenosine - analogs & derivatives Adenosine - pharmacology Adenosine A2 Receptor Agonists - pharmacology Adenosine-5'-(N-ethylcarboxamide) - pharmacology Adult Automation Cell Culture Techniques Cell Proliferation - drug effects Drug Evaluation, Preclinical Erythropoietin - pharmacology Exenatide Female GABA Agents - pharmacology gamma-Aminobutyric Acid - pharmacology Harmine - pharmacology Humans Incretins - pharmacology Insulin-Secreting Cells - drug effects Male Middle Aged Monoamine Oxidase Inhibitors - pharmacology Myostatin - pharmacology Nucleosides - pharmacology Peptides - pharmacology Platelet-Derived Growth Factor - pharmacology Prolactin - pharmacology Regeneration - drug effects Serotonin - pharmacology Serotonin Receptor Agonists - pharmacology Vasodilator Agents - pharmacology Venoms - pharmacology Young Adult |
| Title | Development of a reliable automated screening system to identify small molecules and biologics that promote human β-cell regeneration |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/27624103 https://www.proquest.com/docview/1844028535 |
| Volume | 311 |
| WOSCitedRecordID | wos000390110800007&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| hasFullText | |
| inHoldings | 1 |
| isFullTextHit | |
| isPrint | |
| link | http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwpV1LTsMwELWAIsSGT8un_DRIiF0gSZPYWaEKgVjQigWg7irbsaGoTUqTIvUCHIiDcCbGTirYICGxyc6K44xn5tkz7xFywgQXbiB8J6GedAKa4JZqUeFIL6ZK6zBMLM_24y3tdlmvF99VB255VVY594nWUSeZNGfk54hEEOpgcAkvxq-OUY0yt6uVhMYiqbUwlTFWTXvfbOFG3dXypSLgwrgZzptmQv-cvxiJ2ezMapyYCq_w9xTThprr9f9OcoOsVUkmtEur2CQLKq2TRjtFgD2awSnYsk97nl4nK53qdr1B3n9UEEGmgcNEDQemtwr4FIdiWpoAuhmEvhjwoCSBhiKDge321TPIR3w4hFEpuaty4GkCJc3TQOZQPPMCxrb-T4EVB4TPD8dcHeCLniz_tTGTLfJwfXV_eeNUOg2OxK8tHE69hMXMZTp0NUMIpKmSkeSxq2LBlC8wiTBRUBv6eiEoEzr0eSSly-KABsrfJktplqpdAgiuWrFCSGhY54VApO9rzSgOj7Qfaa9JjucL38d9YGbIU5VN8_730jfJTvn3-uOSsKPvo8cPPLe194fR-2QVrSEq2w0PSE2jF1CHZFm-FYN8cmQNDJ_du84Xdzzd1Q |
| linkProvider | ProQuest |
| openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Development+of+a+reliable+automated+screening+system+to+identify+small+molecules+and+biologics+that+promote+human+%CE%B2-cell+regeneration&rft.jtitle=American+journal+of+physiology%3A+endocrinology+and+metabolism&rft.au=Aamodt%2C+Kristie+I&rft.au=Aramandla%2C+Radhika&rft.au=Brown%2C+Judy+J&rft.au=Fiaschi-Taesch%2C+Nathalie&rft.date=2016-11-01&rft.issn=1522-1555&rft.eissn=1522-1555&rft.volume=311&rft.issue=5&rft.spage=E859&rft_id=info:doi/10.1152%2Fajpendo.00515.2015&rft.externalDBID=NO_FULL_TEXT |
| thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=1522-1555&client=summon |
| thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=1522-1555&client=summon |
| thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=1522-1555&client=summon |