Targeting p38 gamma signaling to advance Cutaneous T Cell Lymphoma Therapy

靶向 p38 γ 信号传导以推进皮肤 T 细胞淋巴瘤治疗

基本信息

批准号：
10296667
负责人：
STEVEN Terry ROSEN
金额：
$ 67.8万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10296667
关键词：
Affect Affinity Animal Model Binding Biological CRISPR screen Cell Death Cell Line Chemicals Clinical Computer Models Computing Methodologies Cutaneous T-cell lymphoma Data Development Drug resistance Elements FDA approved Future Gene Expression Gene Silencing Goals Growth Histone Deacetylase Histone Deacetylase Inhibitor In Vitro Lead Learning Ligands Lymphoma Lymphoma cell Malignant - descriptor Malignant Neoplasms Mediating Methodology Mitogen-Activated Protein Kinases Molecular Organic Synthesis Outcome Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Positioning Attribute Prognosis Proteins RNA interference screen Receptor Signaling Regimen Research Research Personnel Role Sampling Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Site Structure T-Cell Lymphoma T-Cell Receptor T-Lymphocyte Testing Therapeutic Therapeutic Effect Treatment outcome Validation Vorinostat Work Xenograft procedure advanced disease analog base cancer cell cell killing chemical genetics clinical application clinically relevant clinically significant cytotoxic cytotoxicity drug discovery experience genetic approach high throughput screening improved improved outcome in vivo inhibitor innovation knock-down molecular modeling mouse model nanomolar new therapeutic target novel novel therapeutics p38 Mitogen Activated Protein Kinase patient derived xenograft model patient prognosis protein protein interaction response scaffold screening small molecule small molecule inhibitor targeted treatment therapeutically effective tumorigenesis

Affect Affinity Animal Model Binding Biological CRISPR screen Cell Death Cell Line Chemicals Clinical Computer Models Computing Methodologies Cutaneous T-cell lymphoma Data Development Drug resistance Elements FDA approved Future Gene Expression Gene Silencing Goals Growth Histone Deacetylase Histone Deacetylase Inhibitor In Vitro Lead Learning Ligands Lymphoma Lymphoma cell Malignant - descriptor Malignant Neoplasms Mediating Methodology Mitogen-Activated Protein Kinases Molecular Organic Synthesis Outcome Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Positioning Attribute Prognosis Proteins RNA interference screen Receptor Signaling Regimen Research Research Personnel Role Sampling Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Site Structure T-Cell Lymphoma T-Cell Receptor T-Lymphocyte Testing Therapeutic Therapeutic Effect Treatment outcome Validation Vorinostat Work Xenograft procedure advanced disease analog base cancer cell cell killing chemical genetics clinical application clinically relevant clinically significant cytotoxic cytotoxicity drug discovery experience genetic approach high throughput screening improved improved outcome in vivo inhibitor innovation knock-down molecular modeling mouse model nanomolar new therapeutic target novel novel therapeutics p38 Mitogen Activated Protein Kinase patient derived xenograft model patient prognosis protein protein interaction response scaffold screening small molecule small molecule inhibitor targeted treatment therapeutically effective tumorigenesis

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项目摘要

Cutaneous T cell lymphoma (CTCL) is a disfiguring, incurable cancer. For patients with advanced disease, current therapies are inadequate, and outcome is poor. An incomplete understanding of CTCL molecular regulators has limited development of effective targeted therapies. One candidate regulator is p38γ, the gene expression of which is selectively increased in CTCL cell lines and patient samples, but not healthy T cells. We demonstrate that inhibition or silencing of p38γ inhibits proliferation and induces CTCL cell death. The NF-κB pathway is constitutively active in CTCL, provides a complementary T cell signaling pathway to p38γ, and can be inhibited by histone deacetylase inhibitors (HDACi). HDACi, which are currently the most effective clinically approved cytotoxic compounds against CTCL, demonstrate synergistic killing when combined with p38γ Inhibition. Our objective is to understand and exploit the p38γ pathway in CTCL, using a combination of molecular, chemical, and genetic approaches. Our first Aim is to determine the mechanisms by which p38γ inhibition induces cell death in CTCL. We will define the kinase cascade involved in p38γ inhibition-induced CTCL cell killing and identify phosphorylation targets of p38γ signaling; use a synthetic lethal RNAi screen to identify signaling components that cause cell death upon depletion in the presence of p38γ inhibition; and determine the extent to which combined inhibition of p38γ and complementary pathways, including HDACs, induce synergistic therapeutic effects. We will validate identified proteins for the ability to affect downstream signaling and cellular responses in vitro and in vivo, using CTCL cell line xenograft and patient-derived xenograft (PDX) models. Our second Aim is to develop novel p38γ inhibitors for potential therapeutic application. Using high throughput screening and molecular modeling, we identified the multi-kinase inhibitor F7 (also known as PIK75), and showed it is an ATP-competitive p38γ inhibitor with nanomolar cytotoxic efficacy against CTCL cells. To develop a more selective p38γ inhibitor, we will combine ligand- and structure-based computational methods with organic synthesis; using F7 as a scaffold molecule, we will identify F7 analogs and derivatize F7 to have higher a binding affinity for p38γ than other kinases. In addition, we will use CRISPR-based screening to identify novel functional domains and non-conserved sites for developing allosteric next-generation therapeutics. We will synthesize the various analogs and validate hits for CTCL cytotoxicity and p38γ-specific kinase inhibition in vitro and in vivo, using CTCL xenograft and PDX models. We expect that successful completion of this proposal will yield mechanistic information about the unique biological and clinical relevance of p38γ signaling and complementary pathways in CTCL. Importantly, validation of a specific p38γ inhibitor with efficacy in CTCL animal models will have immediate relevance for CTCL therapy.

皮肤T细胞淋巴瘤（CTCL）是一种毁容，无法治愈的癌症。对于患有晚期疾病的患者当前的疗法不足，结果很差。对CTCL分子的不完全理解监管机构的有效靶向疗法的发展有限。一个候选调节剂是p38γ，基因在CTCL细胞系和患者样品中，其表达有选择地增加，但没有健康的T细胞。我们证明p38γ的抑制或沉默会抑制增殖并诱导CTCL细胞死亡。 NF-κB 途径在CTCL中始终处于活动状态，为P38γ提供了完整的T细胞信号传导途径，并且可以由组蛋白脱乙酰基酶抑制剂（HDACI）抑制。 HDACI，目前是临床上最有效的批准的针对CTCL的细胞毒性化合物，与p38γ合并时证明了协同杀伤抑制。我们的目的是结合使用分子，化学和遗传方法。我们的第一个目的是确定p38γ的机制抑制作用诱导CTCL细胞死亡。我们将定义涉及p38γ诱导的激酶级联反应 CTCL细胞杀死并鉴定p38γ信号传导的磷酸化靶标；使用合成致命的RNAi屏幕鉴定在p38γ存在下耗尽时导致细胞死亡的信号传导成分；和确定p38γ和互补途径（包括HDAC，诱导协同理论效应。我们将验证已识别的蛋白质能够影响下游的能力使用CTCL细胞系和患者衍生的Xenogroghothomogonmogroticon和体内信号传导和细胞反应（PDX）模型。我们的第二个目的是开发用于潜在治疗应用的新型p38γ抑制剂。使用高通量筛选和分子建模，我们确定了多激酶抑制剂F7（也称为 PIK75），表明它是一种具有纳摩尔细胞毒性效率的ATP竞争力P38γ抑制剂。为了开发更具选择性的p38γ抑制剂，我们将结合配体和基于结构的计算方法与有机合成；使用F7作为支架分子，我们将识别F7类似物并衍生化F7 对p38γ的结合亲和力高于其他激酶。此外，我们将使用基于CRISPR的筛选来识别用于开发变构的下一代疗法的新型功能领域和非保守部位。我们将合成各种类似物并验证命中的CTCL细胞毒性和p38γ特异性激酶抑制体外和体内，使用CTCL特征和PDX模型。我们预计该提议的成功完成将产生有关p38γ信号和临床相关性和临床相关性的机械信息 CTCL中的完全途径。重要的是，在CTCL中对特定p38γ抑制剂的验证动物模型将立即与CTCL治疗相关。