Advanced High-Content-Screening Applications of Clonogenicity in Cancer

Esquer, Hector; Zhou, Qiong; Abraham, Adedoyin D.; LaBarbera, Daniel V.

doi:10.1177/2472555220926921

Cited by 18 publications

(18 citation statements)

References 28 publications

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“…TCF-driven EMT is linked as a mechanism enabling mesenchymal cells with increased tumorigenic properties, including CSC stemness self-renewal, resistance to apoptosis, and metastatic potential. , This fact is consistent with our published results that isolated M-phenotype tumor cells have significantly increased CSC stemness when compared to other isolated phenotypes . Furthermore, we have reported that CHD1Li 6.0 significantly inhibits CSC stemness, based on the clonogenic colony formation assay. , Thus, we evaluated CHD1Li for their ability to inhibit CSC stemness in SW620 and HCT116 M-phenotype cells, using high-content imaging . CHD1Li effectively inhibited colony formation over a low μM to nM range (Figure A,B).…”

Section: Resultsmentioning

confidence: 99%

Design, Synthesis, and Biological Evaluation of the First Inhibitors of Oncogenic CHD1L

et al. 2022

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Chromodomain helicase DNA-binding protein 1 like (CHD1L) is an oncogene implicated in tumor progression, multidrug resistance, and metastasis in many types of cancer. In this article, we described the optimization of the first lead CHD1L inhibitors (CHD1Li) through drug design and medicinal chemistry. More than 30 CHD1Li were synthesized and evaluated using a variety of colorectal cancer (CRC) tumor organoid models and functional assays. The results led to the prioritization of six lead CHD1Li analogues with improved potency, antitumor activity, and drug-like properties including metabolic stability and in vivo pharmacokinetics. Furthermore, lead CHD1Li 6.11 proved to be an orally bioavailable antitumor agent, significantly reducing the tumor volume of CRC xenografts generated from isolated quasi mesenchymal cells (M-phenotype), which possess enhanced tumorigenic properties. In conclusion, we reported the optimization of first-in-class inhibitors of oncogenic CHD1L as a novel therapeutic strategy with potential for the treatment of cancer.

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Section: Resultsmentioning

confidence: 99%

Design, Synthesis, and Biological Evaluation of the First Inhibitors of Oncogenic CHD1L

et al. 2022

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“…All the chemotherapeutics evaluated decreased the formation of colonies, but doxorubicin, panobinostat, gemcitabine, methotrexate, and cisplatin completely inhibited it; and cyclophosphamide and 5-fluorouracil don't have the capacity to inhibit the CTVT colony formation. These results are relevant because these drugs offer the possibility to fight against stem cells of CTVT that can generate differentiated cancer clones, inducing recurrence or resistance against treatment, representing a complication in the development of effective therapies ( 32 , 33 ). When we analyzed the effect of chemotherapeutics on cell cycle progression in CTVT cells, we determined that different drugs act on subG1 phase (vincristine, panobinostat, gemcitabine, toceranib, cyclophosphamide, and methotrexate).…”

Section: Discussionmentioning

confidence: 99%

In vitro chemosensitivity of a canine tumor venereal transmissible cancer cell line

et al. 2022

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The canine transmissible venereal tumor (CTVT) is the most common malignity in dogs. Because there are reports that this tumor is resistant to vincristine sulfate, the chemotherapeutic options are scarce, and the development of new therapeutic approaches is necessary. In this study, we evaluated the cytotoxic activity of vincristine, doxorubicin, temozolomide, panobinostat, toceranib, gemcitabine, cisplatin, fluorouracil, cyclophosphamide, and methotrexate on a CTVT cell line, determining that all drugs decreased the viability in a dose-dependent manner. Furthermore, they inhibit cellular migration in a time- and drug-dependent manner, as evaluated by the wound healing assay. On the other hand, vincristine, panobinostat, gemcitabine, toceranib, cyclophosphamide, and methotrexate increased the percentage of cells in the subG1 phase, and doxorubicin, temozolomide, gemcitabine, toceranib, and methotrexate decreased the percentage of cells in the synthesis phase. To efficientize the use of vincristine, only toceranib increased the cytotoxic effect of vincristine in a synergistic manner. Our results confirm the use of vincristine as the gold standard for CTVT treatment as monotherapy and suggest the use of a combinatorial and sequential treatment with toceranib.

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“…We again note that this technology does not represent an HTS with a true clonogenic endpoint. A more recent paper reports a High Content Screen (HCS) based on the growth of cancer cells in 2D and in 3D (spheroids) assays ( 8 ). Colony growth of SW620 colorectal cancer cells was monitored following serial seeding dilutions to achieve 1-100 colonies per well, in the presence or absence of drug.…”

Section: Introductionmentioning

confidence: 99%

“…Several recent papers describe methodologies that approximate clonongic assays in HTS formats by miniaturizing to small well formats and utilizing automated image analysis to score fixed and stained colonies (7)(8)(9)(10)(11). The closest approximation of a clonogenic HTS among this body of work is a 'High Content' screen of Non Small Cell Lung Cancer cells in 96-well plates (10).…”

Section: Introductionmentioning

confidence: 99%

A high throughput screen with a clonogenic endpoint to identify radiation modulators of cancer

Gomes

Frederick

Jacobsen

et al. 2022

Preprint

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Clonogenic assays evaluate the ability of single cells to proliferate and form colonies. This process approximates the regrowth and recurrence of tumors after treatment with radiation or chemotherapy, and thereby provides a drug discovery platform for compounds that block this process. However, because of their labor-intensive and cumbersome nature, adapting canonical clonogenic assays for high throughput screening (HTS) has been challenging. We overcame these barriers by developing an integrated system that automates cell- and liquid-handling, irradiation, dosimetry, drug administration, and incubation. Further, we developed a fluorescent live-cell based automated colony scoring methodology that identifies and counts colonies precisely based upon actual nuclei number rather than colony area, thereby eliminating errors in colony counts caused by radiation induced changes in colony morphology. We identified 13 cell lines from 7 cancer types, where radiation is a standard treatment module, that exhibit identical radiation and chemoradiation response regardless of well format and are amenable to miniaturization into small-well HTS formats. We performed pilot screens through a 1584 compound NCI Diversity Set library using two cell lines representing different cancer indications. Radiation modulators identified in the pilot screens were validated in traditional clonogenic assays, providing proof-of-concept for the screen. The integrated methodology, hereafter 'clonogenic HTS', exhibits excellent robustness (Z' values >0.5) and shows high reproducibility (>95%). We propose that clonogenic HTS we developed can function as a drug discovery platform to identify compounds that inhibit tumor regrowth following radiation therapy, to identify new efficacious pair-wise combinations of known oncologic therapies, or to identify novel modulators of approved therapies.

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Advanced High-Content-Screening Applications of Clonogenicity in Cancer

Cited by 18 publications

References 28 publications

Design, Synthesis, and Biological Evaluation of the First Inhibitors of Oncogenic CHD1L

Design, Synthesis, and Biological Evaluation of the First Inhibitors of Oncogenic CHD1L

In vitro chemosensitivity of a canine tumor venereal transmissible cancer cell line

A high throughput screen with a clonogenic endpoint to identify radiation modulators of cancer

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