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BioAssay: AID 1858

Summary of probe development efforts to identify inhibitors of kruppel-like factor 5 (KLF5)

Transcription factors are essential regulators of transcription that bind DNA to control both the rate and frequency of gene expression (1). Many diseases of cell homeostasis are associated with aberrant transcription factor activity (2). Colon cancer, in particular, is a disease of uncontrolled proliferation of the epithelial cells that line the intestinal crypts. Kruppel-like factor 5 (KLF5) is more ..
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AID: 1858
Data Source: The Scripps Research Institute Molecular Screening Center (KLF5_INH_ LEADS_SUMMARY)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-07-09
Modify Date: 2011-10-31
Target
Related Experiments
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AIDNameTypeComment
1700Primary cell-based high throughput screening assay to identify inhibitors of kruppel-like factor 5 (KLF5)Screeningdepositor-specified cross reference: Primary screen (KLF5 inhibitors in singlicate)
1825Luminescence-based counterscreen assay for KLF5 inhibitors: cell-based high throughput screening assay to identify cytotoxic compounds using the IEC-6 intestinal epithelial cell line.Screeningdepositor-specified cross reference: Counterscreen (Cytotoxic compounds in singlicate)
1834Luminescence-based confirmation cell-based high throughput screening assay to identify inhibitors of kruppel-like factor 5 (KLF5)Screeningdepositor-specified cross reference: Confirmation screen (KLF5 inhibitors in triplicate)
1905Luminescence-based counterscreen assay for KLF5 inhibitors: cell-based high throughput screening assay to identify cytotoxic compounds using the IEC-6 intestinal epithelial cell line in triplicate.Screeningdepositor-specified cross reference: Counterscreen (Cytotoxic compounds in triplicate)
1907Luminescence-based confirmation cell-based assay for cytotoxic compounds using the IEC-6 intestinal epithelial cell line.Screeningdepositor-specified cross reference: Confirmation screen (Cytotoxic compounds in triplicate)
1972Luminescence-based dose response cell-based high throughput screening assay for cytotoxic compounds using the IEC-6 intestinal epithelial cell line.Confirmatorydepositor-specified cross reference: Dose response (Cytotoxic compounds in triplicate)
1973Luminescence-based dose response cell-based high throughput screening assay for inhibitors of kruppel-like factor 5 (KLF5).Confirmatorydepositor-specified cross reference: Dose response (KLF5 inhibitors in triplicate)
1975Luminescence-based counterscreen assay for KLF5 inhibitors: dose response cell-based high throughput screening assay to identify cytotoxic compounds using the IEC-6 intestinal epithelial cell line.Confirmatorydepositor-specified cross reference: Dose response counterscreen (Cytotoxic compounds in triplicate)
2749Late stage counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): luminescence-based cell-based dose response assay for cytotoxic compounds using the IEC-6 intestinal epithelial cell lineConfirmatorydepositor-specified cross reference: Late stage counterscreen dose response (Cytotoxic compounds in triplicate)
2750Late stage results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): luminescence-based cell-based dose response assay for inhibitors of KLF5Confirmatorydepositor-specified cross reference: Late stage dose response (KLF5 inhibitors in triplicate)
434956Late stage counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): luminescence-based cell-based dose response assay for cytotoxic compounds using the IEC-6 intestinal epithelial cell line (Round 1)Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (Cytotoxic compounds in triplicate)
434957Late stage results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): luminescence-based cell-based dose response assay for inhibitors of KLF5 (Round 1)Confirmatorydepositor-specified cross reference: Late stage dose response (KLF5 inhibitors in triplicate)
485336Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of KLF5 protein levelsConfirmatorydepositor-specified cross reference: Late stage dose response (KLF5 protein levels inhibitors in triplicate)
485338Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): luminescence-based cell-based dose response assay for cytotoxic compounds using the DLD-1 cell line (Round 1)Confirmatorydepositor-specified cross reference: Late stage dose response counterscreen (Cytotoxicity in triplicate)
588539Late stage assay provider counterscreen results for the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): Cell cycle analysis of the DLD-1 cell lineOtherdepositor-specified cross reference: Late stage counterscreen (Cell cycle analysis of the DLD-1 cell line)
588610Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Epidermal Growth Factor Receptor (EGFR) protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGFR protein levels in DLD1 cells inhibitors)
588612Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Mitogen-Activated Protein Kinase 1 (P-MAPK1; P-ERK1/2) protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-ERK1/2 protein levels in DLD1 cells inhibitors)
588613Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Epidermal Growth Factor Receptor (EGFR) protein levels in DLD1 cells.Otherdepositor-specified cross reference: Late stage counterscreen (P-EGFR protein levels in DLD1 cells inhibitors)
588614Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Mitogen-Activated Protein Kinase 1 (MAPK1; ERK1/2) protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (ERK1/2 protein levels in DLD1 cells inhibitors)
588615Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of P38 protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (P38 protein levels in DLD1 cells inhibitors)
588616Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of phospho-P38 protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (phospho-P38 protein levels in DLD1 cells inhibitors)
588617Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of KLF5 protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (KLF5 protein levels in DLD1 cells inhibitors)
588618Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Early Growth Response 1 (EGR1) protein levels in DLD1 cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGR1 protein levels in DLD1 cells inhibitors)
588640Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of P38 protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P38 protein levels in HCT116 cells inhibitors)
588642Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Mitogen-Activated Protein Kinase 1 (MAPK1; ERK1/2) protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (ERK1/2 protein levels in HCT116 cells inhibitors)
588643Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Mitogen-Activated Protein Kinase 1 (P-MAPK1; P-ERK1/2) protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-ERK1/2 protein levels in HCT116 cells inhibitors)
588644Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Early Growth Response 1 (EGR1) protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGR1 protein levels in HCT116 cells inhibitors)
588646Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of phosphorylated P38 protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (phospho-P38 protein levels in HCT116 cells inhibitors)
588648Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Epidermal Growth Factor Receptor (EGFR) protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGFR protein levels in HCT116 cells inhibitors)
588650Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Epidermal Growth Factor Receptor (EGFR) protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-EGFR protein levels in HCT116 cells inhibitors)
588652Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of KLF5 protein levels in HCT116 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (KLF5 protein levels in HCT116 cells inhibitors)
588653Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Epidermal Growth Factor Receptor (EGFR) protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-EGFR protein levels in HT29 cells inhibitors)
588654Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Epidermal Growth Factor Receptor (EGFR) protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGFR protein levels in HT29 cells inhibitors)
588655Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Mitogen-Activated Protein Kinase 1 (P-MAPK1; P-ERK1/2) protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-ERK1/2 protein levels in HT29 cells inhibitors)
588656Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Mitogen-Activated Protein Kinase 1 (MAPK1; ERK1/2) protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (ERK1/2 protein levels in HT29 cells inhibitors)
588657Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Early Growth Response 1 (EGR1) protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGR1 protein levels in HT29 cells inhibitors)
588658Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of phosphorylated P38 protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (phospho-P38 protein levels in HT29 cells inhibitors)
588659Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of P38 protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P38 protein levels in HT29 cells inhibitors)
588660Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of KLF5 protein levels in HT29 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (KLF5 protein levels in HT29 cells inhibitors)
588661Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of KLF5 protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (KLF5 protein levels in SW620 cells inhibitors)
588662Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of P38 protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P38 protein levels in SW620 cells inhibitors)
588663Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of phosphorylated P38 protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-P38 protein levels in SW620 cells inhibitors)
588665Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Early Growth Response 1 (EGR1) protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGR1 protein levels in SW620 cells inhibitors)
588666Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Mitogen-Activated Protein Kinase 1 (MAPK1; ERK1/2) protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (ERK1/2 protein levels in SW620 cells inhibitors)
588667Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Epidermal Growth Factor Receptor (EGFR) protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (EGFR protein levels in SW620 cells inhibitors)
588668Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Mitogen-Activated Protein Kinase 1 (P-MAPK1; P-ERK1/2) protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-ERK1/2 protein levels in SW620 cells inhibitors)
588677Late stage assay provider counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): chemiluminescence-based western blot assay for inhibitors of Phosphorylated Epidermal Growth Factor Receptor (EGFR) protein levels in SW620 human colorectal carcinoma cellsOtherdepositor-specified cross reference: Late stage counterscreen (P-EGFR protein levels in SW620 cells inhibitors)
588725Late stage counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): Radioactivity-based biochemical assay to identify modulators of a panel of 48 kinasesOtherdepositor-specified cross reference: Late stage counterscreen (modulators of a panel of 48 kinases)
588729Late stage counterscreen results from the probe development effort to identify inhibitors of kruppel-like factor 5 (KLF5): Absorbance-based cell-based assay to identify modulators of cancer cell viabilityConfirmatorydepositor-specified cross reference: Late stage counterscreen (cancer cell viability)
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center
Affiliation: The Scripps Research Institute, TSRI
Assay Provider: Vincent Yang, Emory University
Network: Molecular Library Probe Production Centers Network (MLPCN)
Grant Proposal Number 1-R03-DA026215-01
Grant Proposal PI: Vincent Yang
External Assay ID: KLF5_INH_ LEADS_SUMMARY

Name: Summary of probe development efforts to identify inhibitors of kruppel-like factor 5 (KLF5)

Description:

Transcription factors are essential regulators of transcription that bind DNA to control both the rate and frequency of gene expression (1). Many diseases of cell homeostasis are associated with aberrant transcription factor activity (2). Colon cancer, in particular, is a disease of uncontrolled proliferation of the epithelial cells that line the intestinal crypts. Kruppel-like factor 5 (KLF5) is a zinc finger-containing transcription factor that binds to GC-rich sequences in promoters of numerous genes (3) including cyclin D1 (4), cyclin B1/Cdc2 (4), and integrin-linked kinase (5). KLF5 is highly expressed in rapidly dividing epithelial cells in intestinal crypts (6). This expression pattern of KLF5, along with studies demonstrating that KLF5 mediates the transforming effects of oncogenic H-Ras (7), and that ectopic expression of KLF5 leads to increased cell proliferation and anchorage-independent growth of cultured intestinal epithelial cells (8, 9), suggest that KLF5 may be involved in colon cancer pathogenesis. Therefore, the identification of selective inhibitors of KLF5 may provide useful tools to elucidate the role of KLF5 as a regulator of cellular proliferation and tumor formation in the intestinal epithelium.

Summary of Probe Development Effort:

Following primary HTS in singlicate to identify KLF5 inhibitors (AID1700), counterscreening in singlicate to identify cytotoxic compounds (AID 1825), and confirmation of hit activity in triplicate (AID 1834), certain compounds were identified as possible candidates for probe development. A probe development effort is currently underway at the SRIMSC.

References:
1. Ptashne M. Regulation of transcription: from lambda to eukaryotes. Trends Biochem Sci. 2005 Jun;30(6):275-9.
2. Fre S, Vignjevic D, Schoumacher M, Duffy SL, Janssen KP, Robine S, Louvard D. Adv Cancer Res. 2008;100:85-111. Epithelial morphogenesis and intestinal cancer: new insights in signaling mechanisms.
3. Goldstein BG, Chao HH, Yang Y, Yermolina YA, Tobias JW, Katz JP. Am J Physiol Gastrointest Liver Physiol. 2007 Jun;292(6):G1784-92. Overexpression of Kruppel-like factor 5 in esophageal epithelia in vivo leads to increased proliferation in basal but not suprabasal cells.
4. Ghaleb AM, Nandan MO, Chanchevalap S, Dalton WB, Hisamuddin IM, Yang VW. Kruppel-like factors 4 and 5: the yin and yang regulators of cellular proliferation. Cell Res. 2005 Feb;15(2):92-6.
5. Yang Y, Tetreault MP, Yermolina YA, Goldstein BG, Katz JP. Kruppel-like factor 5 controls keratinocyte migration via the integrin-linked kinase. J Biol Chem. 2008 Jul 4;283(27):18812-20.
6. McConnell BB, Ghaleb AM, Nandan MO, Yang VW. The diverse functions of Kruppel-like factors 4 and 5 in epithelial biology and pathobiology. Bioessays. 2007 Jun;29(6):549-57. Erratum in: Bioessays. 2007 Sep;29(9):946.
7. Nandan MO, Yoon HS, Zhao W, Ouko LA, Chanchevalap S, Yang VW. Kruppel-like factor 5 mediates the transforming activity of oncogenic H-Ras. Oncogene. 2004 Apr 22;23(19):3404-13.
8. Chanchevalap S, Nandan MO, Merlin D, Yang VW. FEBS Lett. 2004 Dec 3;578(1-2):99-105. All-trans retinoic acid inhibits proliferation of intestinal epithelial cells by inhibiting expression of the gene encoding Kruppel-like factor 5.
9. Sun R, Chen X, Yang VW. J Biol Chem. 2001 Mar 9;276(10):6897-900. Epub 2001 Jan 10. Intestinal-enriched Kruppel-like factor (Kruppel-like factor 5) is a positive regulator of cellular proliferation.

Keywords:

Summary AID, KLF5, BTEB2, kruppel-like factor 5, cancer, primary screen, HTS, high throughput screen, 1536, inhibitor, inhibition, luciferase, reporter, luminescence, Scripps, Scripps Florida, The Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Protocol
Please see AIDs 1700, 1825, and 1834 for protocols performed in this probe development effort.
Comment
This probe development project is still underway at the SRIMSC.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML264
PubChem Substance ID (SID) for probe 1: 117686865
PubChem Compound ID (CID) for probe 1: 51003603
Probe type for probe 1: Inhibitor
IC50/EC50 (nM) for probe 1: <350
Target for probe 1: KLF5 (gi: 124263658)
Disease relevance for probe 1: Colon Cancer
Anti-target for probe 1: IEC-6 Cytotoxicity
Fold selectivity for probe 1: >100
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK143546/ (ID: 3036172)
Grant number for probe 1: DA026215-01
PubMed Publication ID (PMID) for probe 1: 21885866
NCBI Book chapter title for probe 1: ML264: An Antitumor Agent that Potently and Selectively Inhibits Krüppel-like Factor Five (KLF5) Expression: A Probe for Studying Colon Cancer Development and Progression
Additional Information
Grant Number: 1-R03-DA026215-01

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