Project description:A BRAF V600E colorectal organoid which is sensitive to MAP kinase inhibition was mutagenised with the chemical mutagen ENU and then drug selected using a combination of Trametinib, Dabrafenib and Cetuximab. Single cell derived organoids were then manually picked and expanded in drug. Resistance was confirmed in a 14 day assay and DNA was collected. These then underwent targeted amplicon-based sequencing to confirm candidate resistance effectors from a screen in 2 2D BRAF V600E colorectal cell lines. Pools of resistant clones were also sequenced.
Project description:We present TORNADO-seq —a high-throughput, a high-content drug discovery platform that for the first time uses next-generation sequencing (NGS) based, targeted RNA-seq in organoids monitoring the expression of 206 genes for the evaluation of complex mixtures of cellular phenotypes. TORNADO-seq is a fast, highly-reproducible, time- and cost-effective (5$ per sample including sequencing cost) method that we used to find drugs that enrich for differentiated cell phenotypes in intestinal organoids. These drugs are highly efficacious against cancer compared to wild type organoids and may therefore become promising candidates in CRC treatment. Further, TORNADO-seq facilitated in-depth insight on the mode of action of these drugs.
Project description:We treated APOB-mutant organoids with different anti-NAFLD drug candidates that were effective at resolving steatosis. To understand the mechanisms of drug action and adverse drug effects we performed bulk RNA-sequencing on drug-treated and vehicle-treated APOB mutant organoids.
Project description:This is a single-centre study based on the Simon 2-stage optimax design: 12 patients will be enrolled initially (Stage I), which will then be expanded to a further 13 patients (Stage II) if 3 or more patients enrolled in stage I of the study achieve an objective response with the chemotherapeutic agent selected by the drug screen assay. A total of 25 patients will be included in both stages of study.
Patients enrolled on study will undergo a fresh biopsy of tumour lesion to obtain cells that will be used to generate patient-derived tumour organoids based on the Invitrocue technology. Organoids will then be subjected to a 10-drug panel screening including: 5-fluorouracil, carboplatin, cyclophosphamide, docetaxel, doxorubicin, gemcitabine, irinotecan, oxaliplatin, paclitaxel and vinorelbine. A further 5 drugs (etoposide, ifosfamide, methotrexate, pemetrexed and topotecan) will be screened if sufficient organoids are grown from the biopsy samples within the screening period.
Physicians will be informed of the results, and choice of chemotherapy will be based on an IRS score of 70% or above. If more than 1 candidate drug with IRS of 70% or above is identified, the physician will exercise his/her discretion to select the most suitable drug based on patient’s comorbidities and organ function.
Project description:aCGH of human melanoma cell lines comparing parental (drug sensitve) vs isogenic drug resistant-derived subline Two condition experiment: two BRAF-V600E mutant cell lines (drug sensitive - parental baseline) vs two derived sublines after chronic exposure to the MEK inhibitor trametinib (drug resistant) are compared
Project description:We obtained small cell lung cancer specimens and normal lung specimens from patients who died of drug-resistant SCLC. The small lung cancer specimens include primary lesions and metastatic lesions. Next generation sequencing was performed to assess the expression of miRNA in drug-resistant small cell lung cancer.
Project description:To identify miRNAs involved in drug resistance of human breast cancer, a miRNA microarray was performed on 5 cases of drug resistant tissues and 5 cases of drug sensitive tissues.The expression levels of totally 2019 miRNAs in 5 pairs of matched, drug resistant and drug sensitive tissues were examined by microarray. There were 27 differentially expressed miRNAs between drug resistant and drug sensitive tissues were identified of which there were 11 significantly up-regulated while the other 16 were down-regulated in drug resistant tissues compared to drug sensitive tissues. It was found that miR-489 was one of the most downregulated miRNAs in drug resistant tissues.