2207 MicroRNA-based single-cell CRISPR screens to discover microRNA-mRNA networks involved in carboplatin resistance in ovarian and lung cancer

卡铂是一种以铂为基础的化疗药物，用于治疗肺癌和卵巢癌。不幸的是，尽管生存率有所提高，但由于卡铂耐药性，许多患者复发并死于疾病。因此，进一步了解耐药性形成的进化过程对于开发能够克服耐药性和癌症复发的有效治疗方法至关重要。MicroRNAs （miRNAs）是通过与mRNA靶点的3'UTR相互作用来抑制转录后基因表达的小RNA分子。研究表明，microrna通常通过调节参与这一过程的蛋白质编码的重要转录本，参与各种癌症的化疗耐药。我们假设miRNA-mRNA相互作用的细胞网络参与了肺癌和卵巢癌的卡铂耐药。最初，为了评估miRNA- mrna相互作用在肺癌和卵巢癌中导致了对卡铂的耐药性，我们将在非耐药和耐药的癌细胞系中整合miRNA和RNA测序。这些方法将发现在化学耐药过程中失调的mirna和潜在靶点。此外，为了了解它们的作用机制，我们将建立第一个基于mirna的单细胞CRISPR筛选，以评估选定的卡铂耐药miRNA-mRNA网络如何在单细胞分辨率下作用于卡铂耐药。重要的miRNA-mRNA相互作用将通过金标准生化实验进一步验证。参与这一过程的mirna靶标网络的发现将为潜在的进化过程提供新的见解，从而为进化原理以及克服肺癌和卵巢癌复发的新治疗途径提供新的见解。

Carboplatin is a platinum-based chemotherapy drug in lung and ovarian cancer treatment. Unfortunately, despite improved survival, numerous patients relapse and succumb to the disease because of carboplatin resistance. Therefore a further understanding of the evolutionary processes underlying resistance formation is crucial for developing effective treatments that are able to overcome resistance and cancer relapse. MicroRNAs (miRNAs) are small RNA molecules that repress post-transcriptional gene expression by interacting with the 3'UTR of mRNA targets. It has been shown that microRNAs are often involved in chemotherapy resistance in various cancers by regulating the important transcripts that code for proteins involved in this process. We hypothesize that cellular networks of miRNA-mRNA interactions are involved in carboplatin resistance in lung and ovarian cancer. Initially, to evaluate which miRNA-mRNA interaction confers resistance to carboplatin in lung and ovarian cancers, we will integrate miRNA and RNA sequencing in non-resistant and resistant cancer cell lines. These approaches will discover the miRNAs and potential targets that are dysregulated during chemoresistance. Furthermore, to understand their mechanism of action we will establish the first miRNA-based single-cell CRISPR screen to evaluate how selected carboplatin-resistance miRNA-mRNA networks act in carboplatin resistance at single-cell resolution. Important miRNA-mRNA interaction will be further validated by performing gold-standard biochemical experiments. The Discovery of miRNA-target networks involved in this process will provide new insights into the underlying evolutionary processes and, hence, into the principles of evolution as well as new therapeutic avenues to overcome lung and ovarian cancer relapse.