دورية أكاديمية
Development and Pilot Screen of Novel High Content Assay for Down Regulators of Expression of Heterogenous Nuclear Ribonuclear Protein H2.
| العنوان: | Development and Pilot Screen of Novel High Content Assay for Down Regulators of Expression of Heterogenous Nuclear Ribonuclear Protein H2. |
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| المؤلفون: | Diez J; Rumbaugh-Goodwin Institute for Cancer Research, Nova Southeastern University, Fort Lauderdale, FL, USA., Rajendrarao S; The Scripps Research Molecular Screening Center, Department of Molecular Medicine, Scripps Research, Jupiter, FL, USA., Baajour SA; Dr. Kiran C. Patel College of Allopathic Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA., Sripadhan P; Rumbaugh-Goodwin Institute for Cancer Research, Nova Southeastern University, Fort Lauderdale, FL, USA., Spicer TP; The Scripps Research Molecular Screening Center, Department of Molecular Medicine, Scripps Research, Jupiter, FL, USA., Scampavia LD; The Scripps Research Molecular Screening Center, Department of Molecular Medicine, Scripps Research, Jupiter, FL, USA., Minond D; Rumbaugh-Goodwin Institute for Cancer Research, Nova Southeastern University, Fort Lauderdale, FL, USA, dminond@nova.edu.; Dr. Kiran C. Patel College of Allopathic Medicine, Nova Southeastern University, Fort Lauderdale, FL, USA. |
| المصدر: | Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology [Cell Physiol Biochem] 2021 May 19; Vol. 55 (3), pp. 265-276. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Physiol Biochem Press GmbH & Co KG Country of Publication: Germany NLM ID: 9113221 Publication Model: Print Cited Medium: Internet ISSN: 1421-9778 (Electronic) Linking ISSN: 10158987 NLM ISO Abbreviation: Cell Physiol Biochem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2019- : Düsseldorf, Germany : Cell Physiol Biochem Press GmbH & Co KG Original Publication: Basel ; New York : S. Karger, 1991-2018. |
| مواضيع طبية MeSH: | Down-Regulation* , Gene Expression Regulation, Neoplastic*, Heterogeneous-Nuclear Ribonucleoprotein Group F-H/*biosynthesis , Melanoma/*metabolism , Neoplasm Proteins/*biosynthesis, Cell Line, Tumor ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H/genetics ; Humans ; Melanoma/genetics ; Melanoma/pathology ; Microscopy, Fluorescence ; Neoplasm Proteins/genetics |
| مستخلص: | Background/aims: Despite recent advances in melanoma drug discovery, the average overall survival of patients with late-stage metastatic melanoma is approximately 3 years, suggesting a need for new approaches and melanoma therapeutic targets. Previously we identified heterogeneous nuclear ribonucleoprotein H2 as a potential target of anti-melanoma compound 2155-14 (Palrasu et al., Cell Physiol Biochem 2019;53:656-686). In the present study, we endeavored to develop an assay to enable a high throughput screening campaign to identify drug-like molecules acting via down regulation of heterogeneous nuclear ribonucleoprotein H2 that can be used for melanoma therapy and research. Methods: We established a cell-based platform using metastatic melanoma cell line WM266-4 expressing hnRNPH2 conjugated with green fluorescent protein to enable assay development and screening. High Content Screening assay was developed and validated in 384 well plate format, followed by miniaturization to 1,536 well plate format. Results: All plate-based QC parameters were acceptable: %CV = 6.7±0.3, S/B = 21±2.1, Z' = 0.75±0.04. Pilot screen of FDA-approved drug library (n=1,400 compounds) demonstrated hit rate of 0.5%. Two compounds demonstrated pharmacological response and were authenticated by western blot analysis. Conclusion: We developed a highly robust HTS-amenable high content screening assay capable of monitoring down regulation of hnRNPH2. This assay is thus capable of identifying authentic down regulators of hnRNPH1 and 2 in a large compound collection and, therefore, is amenable to a large-scale screening effort. Competing Interests: The authors declare that no conflicts of interest exist. (© Copyright by the Author(s). Published by Cell Physiol Biochem Press.) |
| معلومات مُعتمدة: | AR066676, CA249788 United States National Institutes of Health (NIH) |
| فهرسة مساهمة: | Keywords: High content assay; High throughput screening; Heterogenous nuclear ribonuclear protein H; Melanoma |
| المشرفين على المادة: | 0 (HNRNPH2 protein, human) 0 (Heterogeneous-Nuclear Ribonucleoprotein Group F-H) 0 (Neoplasm Proteins) |
| تواريخ الأحداث: | Date Created: 20210520 Date Completed: 20210902 Latest Revision: 20210902 |
| رمز التحديث: | 20240628 |
| DOI: | 10.33594/000000372 |
| PMID: | 34014051 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1421-9778 |
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| DOI: | 10.33594/000000372 |