A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.

Diabetes is a devastating disease that is ultimately caused by the malfunction or loss of insulin-producing pancreatic beta-cells. Drugs capable of inducing the development of new beta-cells or improving the function or survival of existing beta-cells could conceivably cure this disease. We report a...

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Autores principales: Jessica A Hill, Marta Szabat, Corinne A Hoesli, Blair K Gage, Yu Hsuan C Yang, David E Williams, Michael J Riedel, Dan S Luciani, Tatyana B Kalynyak, Kevin Tsai, Ziliang Ao, Raymond J Andersen, Garth L Warnock, James M Piret, Timothy J Kieffer, James D Johnson
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Publicado: Public Library of Science (PLoS) 2010
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spelling oai:doaj.org-article:4597d03eed1c4485a31e9ca5441e029a2021-11-18T06:34:53ZA multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.1932-620310.1371/journal.pone.0012958https://doaj.org/article/4597d03eed1c4485a31e9ca5441e029a2010-09-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20886041/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Diabetes is a devastating disease that is ultimately caused by the malfunction or loss of insulin-producing pancreatic beta-cells. Drugs capable of inducing the development of new beta-cells or improving the function or survival of existing beta-cells could conceivably cure this disease. We report a novel high-throughput screening platform that exploits multi-parameter high-content analysis to determine the effect of compounds on beta-cell survival, as well as the promoter activity of two key beta-cell genes, insulin and pdx1. Dispersed human pancreatic islets and MIN6 beta-cells were infected with a dual reporter lentivirus containing both eGFP driven by the insulin promoter and mRFP driven by the pdx1 promoter. B-score statistical transformation was used to correct systemic row and column biases. Using this approach and 5 replicate screens, we identified 7 extracts that reproducibly changed insulin and/or pdx1 promoter activity from a library of 1319 marine invertebrate extracts. The ability of compounds purified from these extracts to significantly modulate insulin mRNA levels was confirmed with real-time PCR. Insulin secretion was analyzed by RIA. Follow-up studies focused on two lead compounds, one that stimulates insulin gene expression and one that inhibits insulin gene expression. Thus, we demonstrate that multi-parameter, high-content screening can identify novel regulators of beta-cell gene expression, such as bivittoside D. This work represents an important step towards the development of drugs to increase insulin expression in diabetes and during in vitro differentiation of beta-cell replacements.Jessica A HillMarta SzabatCorinne A HoesliBlair K GageYu Hsuan C YangDavid E WilliamsMichael J RiedelDan S LucianiTatyana B KalynyakKevin TsaiZiliang AoRaymond J AndersenGarth L WarnockJames M PiretTimothy J KiefferJames D JohnsonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 9, p e12958 (2010)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Jessica A Hill
Marta Szabat
Corinne A Hoesli
Blair K Gage
Yu Hsuan C Yang
David E Williams
Michael J Riedel
Dan S Luciani
Tatyana B Kalynyak
Kevin Tsai
Ziliang Ao
Raymond J Andersen
Garth L Warnock
James M Piret
Timothy J Kieffer
James D Johnson
A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
description Diabetes is a devastating disease that is ultimately caused by the malfunction or loss of insulin-producing pancreatic beta-cells. Drugs capable of inducing the development of new beta-cells or improving the function or survival of existing beta-cells could conceivably cure this disease. We report a novel high-throughput screening platform that exploits multi-parameter high-content analysis to determine the effect of compounds on beta-cell survival, as well as the promoter activity of two key beta-cell genes, insulin and pdx1. Dispersed human pancreatic islets and MIN6 beta-cells were infected with a dual reporter lentivirus containing both eGFP driven by the insulin promoter and mRFP driven by the pdx1 promoter. B-score statistical transformation was used to correct systemic row and column biases. Using this approach and 5 replicate screens, we identified 7 extracts that reproducibly changed insulin and/or pdx1 promoter activity from a library of 1319 marine invertebrate extracts. The ability of compounds purified from these extracts to significantly modulate insulin mRNA levels was confirmed with real-time PCR. Insulin secretion was analyzed by RIA. Follow-up studies focused on two lead compounds, one that stimulates insulin gene expression and one that inhibits insulin gene expression. Thus, we demonstrate that multi-parameter, high-content screening can identify novel regulators of beta-cell gene expression, such as bivittoside D. This work represents an important step towards the development of drugs to increase insulin expression in diabetes and during in vitro differentiation of beta-cell replacements.
format article
author Jessica A Hill
Marta Szabat
Corinne A Hoesli
Blair K Gage
Yu Hsuan C Yang
David E Williams
Michael J Riedel
Dan S Luciani
Tatyana B Kalynyak
Kevin Tsai
Ziliang Ao
Raymond J Andersen
Garth L Warnock
James M Piret
Timothy J Kieffer
James D Johnson
author_facet Jessica A Hill
Marta Szabat
Corinne A Hoesli
Blair K Gage
Yu Hsuan C Yang
David E Williams
Michael J Riedel
Dan S Luciani
Tatyana B Kalynyak
Kevin Tsai
Ziliang Ao
Raymond J Andersen
Garth L Warnock
James M Piret
Timothy J Kieffer
James D Johnson
author_sort Jessica A Hill
title A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
title_short A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
title_full A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
title_fullStr A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
title_full_unstemmed A multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and Pdx1 expression.
title_sort multi-parameter, high-content, high-throughput screening platform to identify natural compounds that modulate insulin and pdx1 expression.
publisher Public Library of Science (PLoS)
publishDate 2010
url https://doaj.org/article/4597d03eed1c4485a31e9ca5441e029a
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