scholarly journals An Isogenic Cell Line Panel for Sequence-Based Screening of Targeted Anti-Cancer Drugs

2021 ◽  
Author(s):  
Ashley L. Cook ◽  
Nicolas A. Wyhs ◽  
Surojit Sur ◽  
Blair Ptak ◽  
Maria Popoli ◽  
...  
Keyword(s):  
Cell Line ◽  
Cancer Drugs ◽  
Isogenic Cell Line ◽  
Anti Cancer ◽  
Cell Line Panel

scholarly journals An Isogenic Cell Line Panel for Sequence-based Screening of Targeted Anti-cancer Drugs

2021 ◽  
Author(s):  
Ashley L Cook ◽  
Nicolas Wyhs ◽  
Surojit B Sur ◽  
Blair Ptak ◽  
Maria Popoli ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
High Throughput ◽  
High Throughput Screening ◽  
Growth Inhibitor ◽  
Suppressor Gene ◽  
Loss Of Function ◽  
Cancer Pathways ◽  
Isogenic Cell Line ◽  
Cell Line Panel

We describe the creation and characterization of an isogenic cell line panel representing common cancer pathways, with multiple features optimized for high-throughput screening. More than 1,800 cell lines from three normal human cells were generated using CRISPR-technologies. Surprisingly, we discovered most of these lines did not result in complete gene inactivation, despite integration of sgRNA at the desired genomic site. However, a subset of the lines harbored true, biallelic disruptions of the targeted tumor suppressor gene, yielding a final panel of 100 well-characterize lines covering 19 pathways frequently subject to loss of function in cancers. This panel included genetic markers optimized for sequence-based ratiometric assays for drug-based screening assays. To illustrate the potential utility of this panel, we developed a multiplexed high-throughput screen that identified Wee1 inhibitor MK-1775 as a selective growth inhibitor of cells with inactivation of TP53. These cell lines and screening approach should prove useful for researchers studying a variety of cellular and biochemical phenomena.

scholarly journals An isogenic cell line panel identifies major regulators of aberrant astrocyte proliferation in Down syndrome

2020 ◽  
Author(s):  
Keiji Kawatani ◽  
Toshihiko Nambara ◽  
Nobutoshi Nawa ◽  
Hidetaka Yoshimatsu ◽  
Haruna Kusakabe ◽  
...  
Keyword(s):  
Down Syndrome ◽  
Cell Line ◽  
Chromosome 21 ◽  
Small Subset ◽  
Transcriptional Responses ◽  
Genome Chromosome ◽  
Cellular Models ◽  
Isogenic Cell Line ◽  
Cell Line Panel ◽  
Induced Pluripotent

Abstract Astrocytes exert adverse effects on the brains of individuals with Down syndrome (DS). Although a neurogenic-to-gliogenic shift in the fate-specification step has been reported, the mechanisms and key regulators underlying the accelerated proliferation of astrocyte precursor cells (APCs) in DS remain elusive. Here, we established an isogenic cell line panel, based on DS-specific induced pluripotent stem cells, the XIST-mediated transcriptional silencing system in trisomic chromosome 21, and genome/chromosome-editing technologies to eliminate phenotypic fluctuations caused by genetic variation. The transcriptional responses of genes observed upon XIST induction and/or downregulation were not uniform, and only a small subset of genes showed a characteristic expression pattern, which is consistent with the proliferative phenotypes of DS APCs. Comparative analysis and experimental verification using gene modification revealed dose-dependent proliferation-promoting activity of DYRK1A and PIGP on DS APCs. Our collection of isogenic cell lines provides a comprehensive set of cellular models for DS investigations.

Screening of 129 FDA approved anti-cancer drugs in colorectal cancer cell lines resistant to oxaliplatin or irinotecan (SN38).

2017 ◽  
Vol 35 (4_suppl) ◽  
pp. 642-642 ◽  
Author(s):  
Jan Stenvang ◽  
Christine Hjorth Andreassen ◽  
Nils Brünner
Keyword(s):  
Colorectal Cancer ◽  
Drug Resistance ◽  
Cell Viability ◽  
Cell Line ◽  
Cell Lines ◽  
Resistant Cell ◽  
Cancer Drug ◽  
Cancer Drugs ◽  
Drug Candidates ◽  
Anti Cancer

642 Background: In metastatic colorectal cancer (mCRC) only 3 cytotoxic drugs (oxaliplatin, irinotecan and fluorouracil (5-FU)) are approved and the first and second line response rates are about 50% and 10-15%, respectively. Thus, new treatment options are needed. Novel anti-cancer drug candidates are primarily tested in an environment of drug resistance and the majority of novel drug candidates fail during clinical development. Therefore, “repurposing” of drugs has emerged as a promising strategy to apply established drugs in novel indications. The aim of this project was to screen established anti-cancer drugs to identify candidates for testing in mCRC patients relapsing on standard therapy. Methods: We applied 3 parental (drug sensitive) CRC cell lines (HCT116, HT29 and LoVo) and for each cell line also an oxaliplatin and irinotecan (SN38) resistant cell line. We obtained 129 FDA approved anti-cancer drugs from the Developmental Therapeutics Program (DTP) at the National Cancer Institute (NCI) ( https://dtp.cancer.gov/ ). The parental HT29 cell line and the drug resistant sublines HT29-SN38 and HT29-OXPT were exposed to 3 concentrations of each of the anti-cancer drugs. The effect on cell viability was analyzed by MTT assays. Nine of the drugs were analyzed for effect in the LoVo and HCT116 and the SN38- and oxaliplatin-resistant derived cell lines. Results: None of the drugs caused evident differential response between the resistant and sensitive cells or between the SN38 and oxaliplatin resistant cells. The screening confirmed the resistance as the cells displayed resistance to drugs in the same class as the one they were made resistant to. Of the drugs, 45 decreased cell viability in the HT29 parental and oxaliplatin- or SN-38 resistant cell lines. Nine drugs were tested in all nine CRC cell lines and eight decrease cell viability in the nine cell lines. These included drugs in different classes such as epigenetic drugs, antibiotics, mitotic inhibitors and targeted therapies. Conclusions: This study revealed several possible new “repurposing” drugs for CRC therapy, by showing that 45 FDA-approved anti-cancer drugs decrease cell viability in CRC cell lines with acquired drug resistance.

scholarly journals Microfluidic Single-Cell Proteomics Assay Chip: Lung Cancer Cell Line Case Study

Micromachines ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1147
Author(s):  
Yugyung Jung ◽  
Minkook Son ◽  
Yu Ri Nam ◽  
Jongchan Choi ◽  
James R. Heath ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Single Cell ◽  
Cancer Cells ◽  
Cell Line ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Lung Cancer Cell Line ◽  
Cancer Drugs ◽  
Lung Cancer Cell ◽  
Anti Cancer

Cancer is a dynamic disease involving constant changes. With these changes, cancer cells become heterogeneous, resulting in varying sensitivity to chemotherapy. The heterogeneity of cancer cells plays a key role in chemotherapy resistance and cancer recurrence. Therefore, for effective treatment, cancer cells need to be analyzed at the single-cell level by monitoring various proteins and investigating their heterogeneity. We propose a microfluidic chip for a single-cell proteomics assay that is capable of analyzing complex cellular signaling systems to reveal the heterogeneity of cancer cells. The single-cell assay chip comprises (i) microchambers (n = 1376) for manipulating single cancer cells, (ii) micropumps for rapid single-cell lysis, and (iii) barcode immunosensors for detecting nine different secretory and intracellular proteins to reveal the correlation among cancer-related proteins. Using this chip, the single-cell proteomics of a lung cancer cell line, which may be easily masked in bulk analysis, were evaluated. By comparing changes in the level of protein secretion and heterogeneity in response to combinations of four anti-cancer drugs, this study suggests a new method for selecting the best combination of anti-cancer drugs. Subsequent preclinical and clinical trials should enable this platform to become applicable for patient-customized therapies.

scholarly journals Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-Cancer Drugs

2013 ◽  
Vol 51 (6) ◽  
pp. 711-717 ◽  
Author(s):  
Nattapong Puthdee ◽  
Kulthida Vaeteewoottacharn ◽  
Wunchana Seubwai ◽  
Orasa Wonkchalee ◽  
Worasak Kaewkong ◽  
...  
Keyword(s):  
Cell Line ◽  
Cancer Drugs ◽  
In Vivo Screening ◽  
Anti Cancer ◽  
Cholangiocarcinoma Cell

scholarly journals A human isogenic iPSC-derived cell line panel identifies major regulators of aberrant astrocyte proliferation in Down syndrome

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Keiji Kawatani ◽  
Toshihiko Nambara ◽  
Nobutoshi Nawa ◽  
Hidetaka Yoshimatsu ◽  
Haruna Kusakabe ◽  
...  
Keyword(s):  
Down Syndrome ◽  
Cell Line ◽  
Chromosome 21 ◽  
Small Subset ◽  
Transcriptional Responses ◽  
Genome Chromosome ◽  
Cellular Models ◽  
Isogenic Cell Line ◽  
Cell Line Panel ◽  
Induced Pluripotent

AbstractAstrocytes exert adverse effects on the brains of individuals with Down syndrome (DS). Although a neurogenic-to-gliogenic shift in the fate-specification step has been reported, the mechanisms and key regulators underlying the accelerated proliferation of astrocyte precursor cells (APCs) in DS remain elusive. Here, we established a human isogenic cell line panel based on DS-specific induced pluripotent stem cells, the XIST-mediated transcriptional silencing system in trisomic chromosome 21, and genome/chromosome-editing technologies to eliminate phenotypic fluctuations caused by genetic variation. The transcriptional responses of genes observed upon XIST induction and/or downregulation are not uniform, and only a small subset of genes show a characteristic expression pattern, which is consistent with the proliferative phenotypes of DS APCs. Comparative analysis and experimental verification using gene modification reveal dose-dependent proliferation-promoting activity of DYRK1A and PIGP on DS APCs. Our collection of human isogenic cell lines provides a comprehensive set of cellular models for further DS investigations.

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