Targeting Refractory EGFR-Driven Tumors By Induction Of Dominant-Negative EGFR Splicing Variants

通过诱导显性阴性 EGFR 剪接变体靶向难治性 EGFR 驱动的肿瘤

• 批准号: 10062910
• 负责人: Luca Cartegni
• 金额: --
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10062910
• 关键词: Affect; Alternative Splicing; Antibody Response; Antisense Oligonucleotides; Appearance; Automobile Driving; Biodistribution; Biological; Biological Models; Bypass; Cancer Etiology; Cancer Model; Cell Line; Cells; Cessation of life; Chemistry; Clinical Trials; Colorectal Cancer; Data; Dendrimers; Development; Dimerization; Disease Resistance; Dominant-Negative Mutation; Drug Kinetics; Drug Targeting; Drug resistance; EGF gene; EGFR gene; EGFR inhibition; ERBB2 gene; Effectiveness; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Excision; Future; Gene Expression; Generations; Glioblastoma; Heterodimerization; Human; In Vitro; Introns; Investigation; Lead; Length; Ligand Binding; Ligand Binding Domain; Ligands; Malignant Neoplasms; Malignant neoplasm of lung; Methodology; Methods; Modeling; Mutate; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Normal tissue morphology; Oncogenic; Pathologic; Pathway interactions; Pattern; Peptides; Pharmaceutical Preparations; Pharmacology; Phosphotransferases; Play; Polyadenylation; Positioning Attribute; Preparation; Property; Protein Isoforms; Proteins; RNA; RNA Splicing; Receptor Protein-Tyrosine Kinases; Recombinant Proteins; Refractory; Relapse; Residual state; Resistance; Role; STAT3 gene; Safety; Signal Pathway; Signal Transduction; Specificity; Survival Rate; System; Testing; Therapeutic; Time; Toxic effect; Transcript; Transmembrane Domain; Treatment Protocols; Tyrosine Kinase Inhibitor; Variant; Xenograft procedure; autocrine; base; design; drug development; epidermal growth factor receptor VIII; experimental study; extracellular; in vivo; inhibitor/antagonist; knock-down; mouse model; mutant; mutational status; nanoparticle delivery; novel; novel drug class; overexpression; paracrine; programs; receptor; resistance mechanism; success; targeted cancer therapy; tool; tumor; tumorigenesis

ABSTRACT We recently identified a number of novel secreted soluble decoy RTK isoforms (sdRTKs), generated by an alternative splicing/intronic polyadenylation (IPA) mechanism, which can act as potent natural inhibitors of aberrant RTK signaling, a key driving aspect of a large fraction of cancers, including lung cancer. Further, we developed an antisense-based method to effectively induce expression of the inhibitory soluble decoy RTKs in vitro and in vivo. These compounds represent a new class of drugs that carry the advantage of simultaneously knocking down the pathological targets while introducing natural dominant-negative isoforms, thus greatly increasing efficacy. We propose to apply this antisense-based splicing redirection strategy to induce endogenous, soluble decoy EGFR inhibitory variants (sdEGFR) in EGFR-dependent tumors. sdEGFR variants, which are normally expressed at low levels, encode the extracellular ligand-binding domain but lack the intracellular kinase domain. We hypothesize that sdEGFR-inducing compounds inhibit EGFR signaling by a three-fold mechanism: A) removal of full-length EGFR receptor, B) sequestration of EGF and other ligands, and C) neutralizing hetero- dimerization with residual receptors (EGFR, HER2/3, etc.). We further posit that, as the resulting products are secreted, they affect signaling in both targeted and bystander cells alike and interfere with paracrine and autocrine loops, thus exerting an amplified effect. Since its mechanism of action is different from that of current therapies, this strategy should be effective in all refractory tumors where the mechanisms of resistance still relies on EGFR activity, both when such activity is being driven by wild-type EGFR amplification or EGFR-activating mutations. To this end, we will pursue the following specific aims: 1) Characterization of natural inhibitory sdEGFR splicing variants. 2) Optimization of splicing redirection compounds targeted to EGFR. 3) Modulation of endogenous EGFR splicing and polyadenylation in treatment-resistant tumor models. We propose to use non-small cell lung cancer (NSCLC) as our initial cancer model. NSCLC is the most frequent cause of cancer fatalities, with over 130,000 deaths per year, a quarter of which are associated to, and depend on, aberrant EGFR signaling. Notwithstanding the initial efficacy of targeted drugs, such as third-generation tyrosine kinase inhibitor Osimertinib, against most activated EGFR, resistance typically emerges within 2 years. Thus, the development of additional, alternative approaches to treat refractory EGFR-dependent cancers remains an unfulfilled need. Importantly, whereas we propose to use NSCLC as our main model system both in vitro and in vivo, our findings will also directly apply to any cancer where EGFR plays an important role and is frequently mutated or amplified, such as glioblastoma, colorectal cancer and others. Furthermore, analogous compounds can be simply designed against other RTKs or other targets responsible for various mechanisms of drug-resistance and pathological signaling, thus adding a powerful new set of tools to understand such phenomena, and to inform a multitude of therapeutic leads.

抽象的 我们最近确定了许多新型分泌的可溶性诱饵RTK同工型（SDRTK），由替代品生成 剪接/内含子聚腺苷酸化（IPA）机制，可以充当异常RTK信号的有效天然抑制剂，A 包括肺癌在内的大部分癌症的主要驾驶方面。此外，我们开发了一种基于反义的方法 有效地诱导抑制性可溶性诱饵RTK在体外和体内的表达。这些化合物代表 新的药物具有同时击倒病理性靶标的优势的药物 天然主导性同工型，因此大大提高了功效。 我们建议将这种基于反义的剪接重定向策略应用于诱导内源性，可溶性诱饵EGFR EGFR依赖性肿瘤中的抑制变体（SDEGFR）。 SDEGFR变体，通常在低点表达 水平，编码细胞外配体结合结构域，但缺乏细胞内激酶结构域。 我们假设SDEGFR诱导化合物通过三倍的机制抑制EGFR信号传导：a） 去除全长EGFR受体，b）EGF和其他配体的隔离，c）中和杂质 残留受体（EGFR，HER2/3等）二聚化。我们进一步认为，作为生成的产品被分泌， 它们会影响靶向和旁观者细胞的信号传导，并干扰旁分泌和自分泌环， 从而发挥放大作用。由于其作用机制与当前疗法不同，因此该策略 在所有难治性肿瘤中都应有效，因为耐药机制仍然依赖于EGFR活性 当这种活性是由野生型EGFR扩增或EGFR激活突变驱动的。 为此，我们将追求以下具体目标： 1）自然抑制性SDEGFR剪接变体的表征。 2）优化针对EGFR的剪接重定向化合物。 3）调节耐药性肿瘤模型中内源性EGFR剪接和聚腺苷酸化。 我们建议将非小细胞肺癌（NSCLC）用作我们的初始癌症模型。 NSCLC是最常见的原因 癌症死亡人数，每年130,000多人死亡，其中四分之一与异常相关并依赖 EGFR信号传导。尽管有针对性药物的初始功效，例如第三代酪氨酸激酶抑制剂 Osimertinib针对大多数激活的EGFR，通常在2年内出现。因此，发展 治疗难治性EGFR依赖性癌症的其他替代方法仍然是未满足的需求。 重要的是，尽管我们建议在体外和体内使用NSCLC作为我们的主要模型系统，但我们的发现将 还直接适用于EGFR起着重要作用并经常被突变或放大的任何癌症 作为胶质母细胞瘤，大肠癌等。此外，可以简单地设计类似的化合物 RTK或其他负责药物抵抗和病理信号机制的目标，从而增加了 强大的新工具来了解这种现象，并为多种治疗铅提供信息。