دورية أكاديمية
Identifying Small Molecules which Inhibit Autophagy: a Phenotypic Screen Using Image-Based High-Content Cell Analysis.
| العنوان: | Identifying Small Molecules which Inhibit Autophagy: a Phenotypic Screen Using Image-Based High-Content Cell Analysis. |
|---|---|
| المؤلفون: | Peppard JV; Lead Generation and Candidate Realization, R&D, Bridgewater, NJ 07059, U.S.A., Rugg C; Lead Generation and Candidate Realization, R&D, Bridgewater, NJ 07059, U.S.A., Smicker M; Lead Generation and Candidate Realization, R&D, Bridgewater, NJ 07059, U.S.A., Dureuil C; Oncology R&D, Sanofi, 94403, Vitry-Sur-Seine, France., Ronan B; Oncology R&D, Sanofi, 94403, Vitry-Sur-Seine, France., Flamand O; Oncology R&D, Sanofi, 94403, Vitry-Sur-Seine, France., Durand L; Oncology R&D, Sanofi, 94403, Vitry-Sur-Seine, France., Pasquier B; Oncology R&D, Sanofi, 94403, Vitry-Sur-Seine, France. |
| المصدر: | Current chemical genomics and translational medicine [Curr Chem Genom Transl Med] 2014 Feb 07; Vol. 8 (Suppl 1), pp. 3-15. Date of Electronic Publication: 2014 Feb 07 (Print Publication: 2014). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Bentham Open Country of Publication: United Arab Emirates NLM ID: 101616960 Publication Model: eCollection Cited Medium: Print ISSN: 2213-9885 (Print) Linking ISSN: 22139885 NLM ISO Abbreviation: Curr Chem Genom Transl Med Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Sharjah, U.A.E : Bentham Open, [2013]-[2018] |
| مستخلص: | Autophagy plays an important role in cancer and it has been suggested that it functions not only as a tumor suppressor pathway to prevent tumor initiation, but also as a pro-survival pathway that helps tumor cells endure metabolic stress and resist death triggered by chemotherapeutic agents, including acquired resistance. We aimed to identify small-molecule autophagy inhibitors using a HTS/HCA approach through a phenotypic, cell image-based assay, in order to screen multiple biological targets simultaneously and to screen compounds in a physiologically relevant environment. LC3 is a component of the autophagosome, which undergoes a cytoplasmic redistribution from diffuse to punctate dots during autophagy. We employed HeLa cells stably expressing EGFP-LC3 in a primary phenotypic screen. As a first step, a "Validation Library" of about 8,000 pre-selected compounds, about 25% of which had known biological activity and the others representing a range of chemical structures, was run in duplicate both to assess screening suitability and likely hit rate, and to give a valuable preview of possible active structures or biological targets. The primary screen of about 0.25 million compounds yielded around 10,500 positive compounds. These were tested in a suite of further cellular assays designed to eliminate unwanted positives, together with the application of chemi- and bioinformatics to pick out compounds with known biological activity. These processes enabled the selection of compounds that were the most promisingly active and specific. The screening "tree" identified, amongst others with as yet unidentified targets, chemical series active against autophagy-relevant biological targets ULK or Vsp34, validating the phenotypic screening methods selected. Finally, about 400 compounds were fully qualified after following this triage. The development of the assays, compound screening process and the compound triage is described. |
| References: | J Biomol Screen. 1999;4(2):67-73. (PMID: 10838414) Nat Rev Drug Discov. 2011 Jun 24;10(7):507-19. (PMID: 21701501) Autophagy. 2008 Feb;4(2):151-75. (PMID: 18188003) Cell. 2010 Feb 5;140(3):313-26. (PMID: 20144757) Nat Rev Drug Discov. 2006 Dec;5(12):993-6. (PMID: 17139284) Nat Rev Drug Discov. 2012 Sep;11(9):709-30. (PMID: 22935804) N Engl J Med. 2013 Feb 14;368(7):651-62. (PMID: 23406030) Nat Cell Biol. 2013 Jul;15(7):741-50. (PMID: 23685627) Nat Rev Mol Cell Biol. 2011 Dec 14;13(1):7-12. (PMID: 22166994) Nat Chem Biol. 2013 Apr;9(4):206-9. (PMID: 23508174) Proc Natl Acad Sci U S A. 2012 Jul 10;109(28):11178-83. (PMID: 22711801) J Biomol Screen. 2011 Jul;16(6):588-602. (PMID: 21521801) Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):2003-8. (PMID: 22308354) J Cell Sci. 2006 Apr 1;119(Pt 7):1219-32. (PMID: 16522686) |
| فهرسة مساهمة: | Keywords: HTS; LC3; autophagy; high-content screening.; image-based screening; inhibitors; phenotypic screening |
| تواريخ الأحداث: | Date Created: 20140306 Date Completed: 20140305 Latest Revision: 20211021 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC3941084 |
| DOI: | 10.2174/2213988501408010003 |
| PMID: | 24596680 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2213-9885 |
|---|---|
| DOI: | 10.2174/2213988501408010003 |