Divergent Functions of ERK Substrate Binding Domains in Pathogenesis of Myeloproliferative Neoplasms

项目摘要/摘要 Ras/MAPK通路在85%的人类癌症中被激活。然而，针对RAS/MAPK的尝试 信号传递只产生了有限的效果。我们假设这次失败的一个原因是 靶向这一途径是靶向努力集中在这个级联中的激酶的活性部位，这是 未能提供长期的好处。我们假设这种方法的失败是由两个原因造成的 原因：1)活性部位抑制剂，类似于三磷酸腺苷(ATP)，必须克服过度的 在癌细胞中高水平的胞浆ATP；和2)在这个级联中的关键激酶(例如ERK1/2)具有明显的 底物相互作用结构域可以在调节癌症进展中发挥拮抗作用。 因此，活性部位的集中抑制类似于同时抑制油门和刹车。 汽车的踏板。我们假设，更成功的方法将是开发出 保持激酶活性，但只将其转移到“刹车踏板”底物上。作为支持，我们已经证明了 ERK2的两个底物结合域，称为D和DPB结构域，在 JAK2-激酶驱动的骨髓增生性肿瘤的发病机制。事实上，DBP和D结构域起作用 比如刹车踏板和油门踏板，分别起到了对抗和促进疾病发展的作用。因此， 加速器踏板(D区)或其底物的药物衰减应阻碍肿瘤进展 比活性部位抑制更有效，因为它选择性地干扰了 ERK2，同时保留了DBP结构域的肿瘤抑制功能。虽然我们有令人信服的证据 对于ERK2-D和DBP结构域的相反作用，其作用的分子基础尚不清楚 这是开发有效的、专注于ERK2D结构域的药物干预的障碍。 我们现在寻求根据以下目标解决这一知识差距。我们将：1)评估一般性 ERK2-D和DBP结构域在不同MPN亚型发病机制中的相反作用；2) 了解ERK2-D和DBP结构域发挥不同功能的机制基础；以及3) 评价ERK2-D结构域和/或靶点的药物靶向抗癌效果 进步。通过这些努力，我们希望为ERK2底物结合的作用带来新的见解 调节癌症进展的模块以及如何在治疗上利用这些信息。当我们开始的时候 MPN，这些发现可能对其他RAS/MAPK驱动的癌症有深远的影响。

PROJECT SUMMARY/ABSTRACT The Ras/MAPK pathway is activated in 85% of human cancer. Nevertheless, attempts to target Ras/MAPK signaling have produced only limited efficacy. We hypothesize that one reason for the failure to successfully target this pathway is that the targeting efforts have focused on the active sites of kinases in this cascade, which has failed to provide long-lasting benefit. We hypothesize that the failure of this approach results from two causes: 1) Active site inhibitors, which resemble adenosine triphosphate (ATP), must overcome the exceedingly high cytosolic ATP levels in cancer cells; and 2) critical kinases in this cascade (e.g., ERK1/2) have distinct substrate interactions domains that can perform antagonistic roles in regulating cancer progression. Consequently, active-site focused inhibition is akin to simultaneous depression of the accelerator and brake pedals of an automobile. We hypothesize that a more successful approach will be to develop inhibitors which preserve kinase activity, but divert it exclusively to “brake pedal” substrates. In support, we have demonstrated that the two substrate binding domains of ERK2, termed the D and DPB domains, play opposing roles in the pathogenesis of JAK2-kinase driven myeloproliferative neoplasms (MPN). Indeed, the DBP and D domains act like brake and accelerator pedals, opposing and promoting disease progression, respectively. Consequently, pharmacologic attenuation of the accelerator pedal (D-domain) or its substrates should impair tumor progression more potently than active site inhibition, because it selectively interferes with the disease promoting activity of ERK2, while preserving the tumor suppressive function of the DBP-domain. While we have compelling evidence for the opposing roles of the ERK2-D and DBP domains, the molecular basis for their action remains unclear and this is an impediment to developing effective, pharmacologic interventions focused on the ERK2 D domain. We now seek to address this gap in knowledge according to the following aims. We will: 1) Assess the generality of the opposing functions of the ERK2-D and DBP domains in the pathogenesis of distinct MPN subtypes; 2) Understand the mechanistic basis by which the ERK2-D and DBP domains exert their distinct functions; and 3) Assess the efficacy of pharmacologic targeting of the ERK2-D domain and/or its targets in inhibiting cancer progression. Through these efforts we expect to bring new insights into the role of ERK2 substrate binding modules in regulating cancer progression and how to exploit this information therapeutically. While we begin with MPN, these findings may have far reaching implications for other Ras/MAPK driven cancers.