Large-Scale Small-Molecule Screen Using Zebrafish Embryos

Abstract INTRODUCTION Glioblastoma multiforme (GBM) is the most aggressive form of primary brain cancer. A subpopulation of multipotent cells termed GBM cancer stem cells (CSCs) play a critical role in tumor initiation and maintenance, drug resistance, and recurrence following surgery. New therapeutic strategies for the treatment of GBM have recently focused on targeting CSCs. Here we have used an unbiased large-scale screening approach to identify drug-like small molecules that induce apoptosis in GBM CSCs in a cell type-selective manner. METHODS A luciferase-based survival assay of patient-derived GBM CSC lines was established to perform a large-scale screen of ∼one million drug-like small molecules with the goal of identifying novel compounds that are selectively toxic to chemoresistant GBM CSCs. Compounds found to kill GBM CSC lines as compared to control cell types were further characterized. A caspase activation assay was used to evaluate the mechanism of induced cell death. A xenograft animal model using patient-derived GBM CSCs was employed to test the leading candidate for suppression of in vivo tumor formation. RESULTS We identified a small molecule, termed RIPGBM, from the cell-based chemical screen that induces apoptosis in primary patient-derived GBM CSC cultures. The cell type-dependent selectivity of RIPGBM appears to arise at least in part from redox-dependent formation of a proapoptotic derivative, termed cRIPGBM, in GBM CSCs. cRIPGBM induces caspase 1-dependent apoptosis by binding to receptor-interacting protein kinase 2 (RIPK2) and acting as a molecular switch, which reduces the formation of a prosurvival RIPK2/TAK1 complex and increases the formation of a proapoptotic RIPK2/caspase 1 complex. In an intracranial GBM xenograft mouse model, RIPGBM was found to significantly suppress tumor formation. CONCLUSIONS Our chemical genetics-based approach has identified a small molecule drug candidate and a potential drug target that selectively targets cancer stem cells and provides an approach for the treatment of GBMs.

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Small-Molecule Screen Identifies De Novo Nucleotide Synthesis as a Vulnerability of Cells Lacking SIRT3

Cell Reports ◽

10.1016/j.celrep.2018.01.076 ◽

2018 ◽

Vol 22 (8) ◽

pp. 1945-1955 ◽

Cited By ~ 7

Author(s):

Karina N. Gonzalez Herrera ◽

Elma Zaganjor ◽

Yoshinori Ishikawa ◽

Jessica B. Spinelli ◽

Haejin Yoon ◽

...

Keyword(s):

Small Molecule ◽

De Novo ◽

Nucleotide Synthesis ◽

Small Molecule Screen

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Small molecule screen for compounds that affect vascular development in the zebrafish retina

Mechanisms of Development ◽

10.1016/j.mod.2009.01.002 ◽

2009 ◽

Vol 126 (5-6) ◽

pp. 464-477 ◽

Cited By ~ 68

Author(s):

Satish S. Kitambi ◽

Kyle J. McCulloch ◽

Randall T. Peterson ◽

Jarema J. Malicki

Keyword(s):

Small Molecule ◽

Vascular Development ◽

Small Molecule Screen

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A small molecule screen identified N-Acetyl-L-cysteine for promoting ex vivo growth of single circulating tumor cells

10.1101/2020.08.19.257378 ◽

2020 ◽

Author(s):

Teng Teng ◽

Mohamed Kamal ◽

Oihana Iriondo ◽

Yonatan Amzaleg ◽

Chunqiao Luo ◽

...

Keyword(s):

Tumor Cells ◽

Circulating Tumor Cells ◽

Small Molecule ◽

Ex Vivo ◽

Single Cells ◽

Small Subset ◽

Blood Draw ◽

Small Molecule Screen ◽

Functional Analyses ◽

Single Ctcs

AbstractCirculating tumor cells (CTCs) can be isolated via a minimally invasive blood draw and are considered a “liquid biopsy” of their originating solid tumors. CTCs contain a small subset of metastatic precursors that can form metastases in secondary organs, and provide a resource to identify mechanisms underlying metastasis-initiating properties. Despite technological advancements that allow for highly sensitive approaches of detection and isolation, CTCs are very rare and often present as single cells, posing an extreme challenge for ex vivo expansion after isolation. Here, using previously established patient-derived CTC lines, we performed a small molecule drug screening to identify compounds that can improve ex vivo culture efficiency for single CTCs. We found that N-acetylcysteine (NAC) and other antioxidants can promote ex vivo expansion of single CTCs, by reducing oxidative and other stress particularly at the initial stage of single cell expansion. RNA-seq analysis of growing clones and non-growing clones confirmed the effect by NAC, but also indicate that NAC-induced decrease in oxidative stress is insufficient for promoting proliferation of a subset of cells with heterogeneous quiescent and senescent features. Despite the challenge in expanding all CTCs, NAC treatment lead to establishment of single CTC clones that have similar tumorigenic features, which will facilitate future functional analyses.

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Enabling Large-Scale Design, Synthesis and Validation of Small Molecule Protein-Protein Antagonists

PLoS ONE ◽

10.1371/journal.pone.0032839 ◽

2012 ◽

Vol 7 (3) ◽

pp. e32839 ◽

Cited By ~ 79

Author(s):

David Koes ◽

Kareem Khoury ◽

Yijun Huang ◽

Wei Wang ◽

Michal Bista ◽

...

Keyword(s):

Small Molecule ◽

Large Scale ◽

Design Synthesis ◽

Scale Design

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Sustainable, Rapid Synthesis of Bright-Luminescent CuInS2-ZnS Alloyed Nanocrystals: Multistage Nano-xenotoxicity Assessment and Intravital Fluorescence Bioimaging in Zebrafish-Embryos

Scientific Reports ◽

10.1038/srep26078 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 21

Author(s):

S. Shashank Chetty ◽

S. Praneetha ◽

Sandeep Basu ◽

Chetana Sachidanandan ◽

A. Vadivel Murugan

Keyword(s):

Large Scale ◽

Near Infrared ◽

Organic Dyes ◽

Low Cost ◽

Fluorescence Emission ◽

Zebrafish Embryos ◽

Fluorescence Bioimaging ◽

Rapid Labeling

Abstract Near-infrared (NIR) luminescent CuInS2-ZnS alloyed nanocrystals (CIZS-NCs) for highly fluorescence bioimaging have received considerable interest in recent years. Owing, they became a desirable alternative to heavy-metal based-NCs and organic dyes with unique optical properties and low-toxicity for bioimaging and optoelectronic applications. In the present study, bright and robust CIZS-NCs have been synthesized within 5 min, as-high-as 230 °C without requiring any inert-gas atmosphere via microwave-solvothermal (MW-ST) method. Subsequently, the in vitro and in vivo nano-xenotoxicity and cellular uptake of the MUA-functionalized CIZS-NCs were investigated in L929, Vero, MCF7 cell lines and zebrafish-embryos. We observed minimal toxicity and acute teratogenic consequences upto 62.5 μg/ml of the CIZS-NCs in zebrafish-embryos. We also observed spontaneous uptake of the MUA-functionalized CIZS-NCs by 3 dpf older zebrafish-embryos that are evident through bright red fluorescence-emission at a low concentration of 7.8 μg/mL. Hence, we propose that the rapid, low-cost, large-scale “sustainable” MW-ST synthesis of CIZS-NCs, is an ideal bio-nanoprobe with good temporal and spatial resolution for rapid labeling, long-term in vivo tracking and intravital-fluorescence-bioimaging (IVBI).

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