Preventing adaptive drug resistance through Mediator kinase inhibition

通过抑制介体激酶来预防适应性耐药性

基本信息

批准号：
10668528
负责人：
Eugenia V Broude
金额：
$ 59.69万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10668528
关键词：
Affect Antineoplastic Agents Breast Cancer cell line Breast Carcinoma Cell Line Cells Derivation procedure Development Drug Targeting Drug resistance ERBB2 gene Epithelium Exposure to Gene Expression Genetic Transcription Genomics Growth Heterogeneity Human Human Cell Line Immune Immune system Immunocompetent Immunodeficient Mouse Immunologic Deficiency Syndromes In Vitro Malignant Epithelial Cell Mediator Mesenchymal MicroRNAs Mouse Mammary Tumor Virus Mus Mutagenesis Neoplasm Metastasis Paclitaxel Pathway interactions Patient-derived xenograft models of breast cancer Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Phosphorylation Phosphotransferases Population Prevention Process Property Protein Isoforms Protein Kinase Remission Induction Resistance Resistance development Role Signal Transduction Signal Transduction Pathway Stromal Cells Testing Treatment outcome Tumor Promotion Xenograft procedure acquired drug resistance cancer drug resistance cancer stem cell cancer therapy clinical development clinically relevant exome sequencing gene induction genetically modified cells in vivo inhibitor inhibitor therapy kinase inhibitor lapatinib malignant breast neoplasm miRNA expression profiling mouse model mutant neoplastic cell non-genetic novel novel drug class novel therapeutics patient derived xenograft model pharmacologic pre-clinical prevent programs prototype single-cell RNA sequencing therapy development therapy resistant transcription factor transcriptional reprogramming transcriptome sequencing transcriptomics tumor tumor growth tumor initiation tumor progression tumor xenograft

Affect Antineoplastic Agents Breast Cancer cell line Breast Carcinoma Cell Line Cells Derivation procedure Development Drug Targeting Drug resistance ERBB2 gene Epithelium Exposure to Gene Expression Genetic Transcription Genomics Growth Heterogeneity Human Human Cell Line Immune Immune system Immunocompetent Immunodeficient Mouse Immunologic Deficiency Syndromes In Vitro Malignant Epithelial Cell Mediator Mesenchymal MicroRNAs Mouse Mammary Tumor Virus Mus Mutagenesis Neoplasm Metastasis Paclitaxel Pathway interactions Patient-derived xenograft models of breast cancer Patients Pharmaceutical Preparations Pharmacotherapy Phenotype Phosphorylation Phosphotransferases Population Prevention Process Property Protein Isoforms Protein Kinase Remission Induction Resistance Resistance development Role Signal Transduction Signal Transduction Pathway Stromal Cells Testing Treatment outcome Tumor Promotion Xenograft procedure acquired drug resistance cancer drug resistance cancer stem cell cancer therapy clinical development clinically relevant exome sequencing gene induction genetically modified cells in vivo inhibitor inhibitor therapy kinase inhibitor lapatinib malignant breast neoplasm miRNA expression profiling mouse model mutant neoplastic cell non-genetic novel novel drug class novel therapeutics patient derived xenograft model pharmacologic pre-clinical prevent programs prototype single-cell RNA sequencing therapy development therapy resistant transcription factor transcriptional reprogramming transcriptome sequencing transcriptomics tumor tumor growth tumor initiation tumor progression tumor xenograft

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项目摘要

Cancer drug resistance is driven in part by the plasticity of tumor cells that allows for therapy-induced adaptation of their transcriptional program. This adaptive drug resistance is associated with the acquisition of various phenotypic changes that promote tumor growth, metastasis and resistance to other therapies. Experimental drugs targeting CDK8/19 Mediator kinase that regulates transcriptional reprogramming were found to suppress the development of resistance to different classes of targeted and conventional anticancer agents. We hypothesize that the emergence of adaptive drug resistance in vitro and in vivo, with concurrent acquisition of tumor-promoting phenotypes, can be prevented by inhibiting Mediator kinase. We will test this hypothesis in HER2-positive breast cancers by analyzing the effects of Mediator kinase inhibition on the emergence of resistance to a HER2-targeting drug (lapatinib) and a conventional drug (paclitaxel). We will pursue the following Specific Aims. (1) The effect of Mediator kinase deficiency on the development of adaptive drug resistance in vitro will be analyzed by generating derivatives of HER2-positive human breast cancer cell lines that will express wild-type or kinase-dead Mediator kinase and by analyzing the effects of Mediator kinase mutagenesis or treatment with selective CDK8/19 inhibitors on the emergence of adaptive resistance to lapatinib or paclitaxel. Comprehensive phenotypic, genomic and transcriptomic analyses will be used to evaluate the effect of drug adaptation and Mediator kinase inhibition on the acquisition of tumor-promoting phenotypes and to identify signal transduction pathways, inhibitors of which could be combined with CDK8/19 inhibitors to enhance the prevention of resistance. (2) HER2-positive human cell lines with different Mediator kinase status and a panel of HER2- positive breast cancer patient-derived xenografts (PDX) will be selected for lapatinib and paclitaxel resistance in vivo, to compare the resistance-preventing effects of Mediator kinase deficiency in tumor cells alone and with the effects of its pharmacological inhibition in both tumor and stromal cells. Transcriptomic and phenotypic analysis will be used to delineate the effects of drug selection and Mediator kinase inhibition on gene expression in tumor and stromal cells. Whole exome sequencing of PDX tumors will reveal if Mediator kinase inhibition prevents the emergence of new drug-resistant tumor lineages or suppresses the growth of drug-resistant cells that may pre-exist in heterogeneous PDXs. (3) The effects of Mediator kinase on the emergence of drug resistance will also be investigated in a murine Her2/Neu-driven mouse mammary carcinoma cells adapted for syngeneic growth, both in vitro (as in Aim 1) and in vivo using both immunocompetent and immunodeficient hosts. This analysis will elucidate the effect of the host immune system on drug adaptation and the role of Mediator kinase in this effect. The proposed program will delineate the roles of Mediator kinase in tumor adaptation to treatment, in vitro and in vivo, and will indicate whether Mediator kinase-inhibiting drugs may be able to extend the duration of remission induced by cancer therapy.

癌症耐药性的一部分是由肿瘤细胞的可塑性驱动的，肿瘤细胞允许治疗诱导的适应他们的转录程序。这种自适应耐药性与获取各种促进肿瘤生长，转移和对其他疗法的耐药性的表型变化。实验靶向调节转录重编程的CDK8/19介质激酶的药物可抑制对不同类别的靶向和常规抗癌药的抵抗力的发展。我们假设体外和体内自适应耐药性的出现，并同时获得可以通过抑制介体激酶来预防促肿瘤的表型。我们将在 HER2阳性乳腺癌通过分析介体激酶抑制对出现的影响对HER2靶向药物（Lapatinib）和常规药物（紫杉醇）的抗性。我们将追求以下具体目标。（1）介质激酶缺乏症对自适应耐药性发展的影响体外将通过产生HER2阳性人类乳腺癌细胞系的衍生物来分析，这些细胞将表达野生型或激酶死亡介质激酶，并通过分析介体激酶诱变或选择性CDK8/19抑制剂对Lapatinib或紫杉醇的适应性抗性进行治疗。全面的表型，基因组和转录组分析将用于评估药物的作用适应和介体激酶抑制对促进肿瘤表型的获取并识别信号的抑制转导途径，其抑制剂可以与CDK8/19的抑制剂结合起来，以增强预防阻力。（2）具有不同介体激酶状态的HER2阳性人体细胞系和HER2-面板乳腺癌阳性患者衍生异种移植（PDX）将在拉帕替尼和紫杉醇耐药性中选择体内，比较单独和与其药理抑制在肿瘤和基质细胞中的作用。转录组和表型分析将用于描述药物选择和介体激酶抑制对基因表达的影响在肿瘤和基质细胞中。 PDX肿瘤的整个外显子组测序将揭示介质激酶抑制作用是否防止抗药性肿瘤谱系的出现或抑制耐药细胞的生长这可能是在异质PDXs中先前存在的。（3）介质激酶对药物出现的影响还将在适应的鼠类HER2/NEU驱动的小鼠乳腺癌细胞中研究抗性合成性生长，无论是在体外（AIM 1）还是使用免疫能力和免疫缺陷的体内生长主持人。该分析将阐明宿主免疫系统对药物适应的影响和介体激酶在这种作用中。拟议的程序将描述介质激酶在肿瘤中的作用适应治疗，体外和体内，将表明抑制介质激酶的药物是否可能是能够延长癌症治疗引起的缓解持续时间。