Targeting DUSP-5 to Treat Vascular Anomalies

靶向 DUSP-5 治疗血管异常

• 批准号: 8259361
• 负责人: Ramani Ramchandran
• 金额: $ 50.59万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-21 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8259361
• 关键词: Affect; Affinity; Alternative Therapies; Basic Science; Biochemical; Biological Assay; Birth; Blood Vessels; C-terminal; Cell Proliferation; Cell physiology; Cells; Cellular biology; Chemicals; Clinical; Common Neoplasm; Computer Simulation; Data; Development; Developmental Biology; Disease; Docking; Embryo; Endothelial Cells; Enzymes; FDA approved; Family; Goals; Health; Hemangioma; Human; In Vitro; Inborn Genetic Diseases; Individual; Infant; Inhibitory Concentration 50; Lead; Life; MAPK3 gene; Mission; Mitogen-Activated Protein Kinases; Modification; Molecular; Molecular Biology; Mutate; Mutation; PP5 protein-serine-threonine phosphatase; Pathogenesis; Patients; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Positioning Attribute; Proliferating; Proline; Protein Kinase; Proteins; Public Health; Research; Serine; Signal Transduction; Specificity; Structure; Suramin; Testing; Therapeutic; Translating; United States National Institutes of Health; Vascular Endothelial Growth Factors; Work; Zebrafish; analog; base; design; extracellular; improved; in vitro Assay; in vivo; infancy; innovation; malformation; member; mutant; novel therapeutics; protein function; small molecule; therapeutic target; tumor growth

DESCRIPTION (provided by applicant): Vascular anomalies (VAs), inborn errors in embryonic vascular development are classified into two distinct groups: hemangiomas and vascular malformations (VMs). Current therapies for VAs are limited in efficacy and have significant complications. Therefore, to improve therapy for patients afflicted with these conditions, it is critical to find new drugs or repurpose FDA-approved drugs to target VAs. Our long-term goal is to understand the underlying mechanisms that lead to pathogenesis of VAs so that better therapeutics targeting this condition can be generated. In order to pursue that goal, the objective is to identify small molecules (SMs) that will target dual-specific phosphatase-5 (Dusp-5), a member of the mitogen-activated protein kinase (MAPK) family, which is mutated in patients with hemangiomas and VMs. We have identified a serine to proline mutation at 147 AA in DUSP-5 (S147P), which results in a hypoactive phosphatase that is unable to dephosphorylate p-ERK. This results in sustained p-ERK levels, which is often associated with increased proliferation of cells such as those in VAs. Our central hypothesis is that, "human S147P protein recapitulates zebrafish S148P protein function, whereby mutation perturbs the interaction with p-ERK such that DUSP5 phosphatase domain (PD) cannot be properly positioned to de-phosphorylate p-ERK. Small molecules such as SM1842, FDA-approved compounds (Suramin), and SM1842 analogs can reverse this effect, thereby permitting a switch between WT and S147P function both at the molecular level (in vitro), and in terms of cellular function." This hypothesis is formulated based on preliminary data from our group that predicts the incorrect positioning of the DUSP5 PD domain in relation to p-ERK using computational modeling studies on Dusp-5 interaction with ERK, which suggests the molecular mechanism that leads to the S147P<s hypoactivity. Further, computational docking approach with 10,500 SM compounds to the C-terminal PD of Dusp-5 enzyme identified SM1842, and other SMs that act as potent Dusp-5 antagonist in ECs. SM1842 is the most potent of the identified hits, acts as Dusp-5 antagonist in p-ERK assay, affects VEGF-stimulated p-ERK and Dusp-5 levels in endothelial cells, and restores S148P function in biochemical assays in vitro. Suramin - FDA-approved compounds (similar structure to SM1842) also affect endogenous Dusp-5 and p-ERK levels in endothelial cells. The proposed hypothesis will be tested by pursuing three specific aims: 1) Determine the structural mechanism for SM1842 in affecting Dusp-5 and S147P function; 2) Identify the optimal chemical analog for SM1842; and 3) Characterize the activity of FDA- approved compounds (suramin) similar to SM1842 in vivo and in vitro. In each of these aims, we will employ a variety of biophysical, cell biology, molecular and developmental biology approaches to unravel mechanistic basis for SM1842 and its analog to affect Dusp-5<s activity in vivo and in vitro. The approach is innovative because molecules like SM1842 that selectively affect mutant protein over WT protein, function are highly sought after by big Pharma, and this application has the potential to shift paradigm in target-based research. The proposed research is significant because benefits of this project will provide immediate clinical impact for VA patients in terms of therapy options, and importantly will translate basic science discovery into tangible clinical benefits instantaneously. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because vascular anomalies (VAs) represent an important clinical problem that has few therapeutic options. The successful development of a small molecule or FDA- approved compound that targets mutated Dusp-5 over WT Dusp-5 protein will provide the much needed therapy alternatives for patients with VAs. Thus, the proposed research is directly relevant to NIH<s mission of reducing the burden of debilitating health conditions from diseases affected by deregulated vasculature.

描述（由申请人提供）：血管异常（VA），胚胎血管发育中的先天性缺陷分为两个不同的组：血管瘤和血管畸形（VM）。目前的 VA 疗法疗效有限，并且有严重的并发症。因此，为了改善患有这些疾病的患者的治疗，寻找新药或重新利用 FDA 批准的药物来靶向 VA 至关重要。我们的长期目标是了解导致 VA 发病机制的潜在机制，以便针对这种情况产生更好的治疗方法。为了实现这一目标，我们的目标是确定靶向双特异性磷酸酶 5 (Dusp-5) 的小分子 (SM)，双特异性磷酸酶 5 是丝裂原激活蛋白激酶 (MAPK) 家族的成员，该家族在血管瘤和 VM 患者中发生突变。我们在 DUSP-5 (S147P) 的 147 AA 处发现了丝氨酸到脯氨酸的突变，该突变导致磷酸酶活性低下，无法使 p-ERK 去磷酸化。这导致 p-ERK 水平持续升高，这通常与细胞增殖增加相关，例如 VA 中的细胞增殖。我们的中心假设是，“人类 S147P 蛋白概括了斑马鱼 S148P 蛋白的功能，突变扰乱了与 p-ERK 的相互作用，使得 DUSP5 磷酸酶结构域 (PD) 无法正确定位以对 p-ERK 进行去磷酸化。小分子，例如 SM1842、FDA 批准的化合物（苏拉明）和 SM1842 类似物可以逆转这种效应，从而允许在分子水平（体外）和细胞功能方面在 WT 和 S147P 功能之间进行切换。”这一假设是基于我们小组的初步数据提出的，该数据利用 Dusp-5 与 ERK 相互作用的计算模型研究预测了 DUSP5 PD 结构域相对于 p-ERK 的错误定位，这表明了导致 S147P<s 活性减退的分子机制。此外，使用 10,500 个 SM 化合物与 Dusp-5 酶 C 端 PD 的计算对接方法鉴定出 SM1842 和其他在 EC 中充当有效 Dusp-5 拮抗剂的 SM。 SM1842 是已确定的最有效的命中，在 p-ERK 测定中充当 Dusp-5 拮抗剂，影响内皮细胞中 VEGF 刺激的 p-ERK 和 Dusp-5 水平，并在体外生化测定中恢复 S148P 功能。 Suramin - FDA 批准的化合物（与 SM1842 结构相似）也会影响内皮细胞中的内源性 Dusp-5 和 p-ERK 水平。所提出的假设将通过三个具体目标进行检验：1）确定SM1842影响Dusp-5和S147P功能的结构机制； 2) 确定 SM1842 的最佳化学类似物； 3) 表征 FDA 批准的化合物（苏拉明）与 SM1842 相似的体内和体外活性。在每个目标中，我们将采用各种生物物理、细胞生物学、分子和发育生物学方法来揭示 SM1842 及其类似物影响 Dusp-5<s 体内和体外活性的机制基础。该方法具有创新性，因为像 SM1842 这样选择性影响突变蛋白而不是 WT 蛋白的分子受到大型制药公司的高度追捧，并且该应用有可能改变基于目标的研究范式。拟议的研究意义重大，因为该项目的好处将为 VA 患者的治疗选择提供直接的临床影响，更重要的是，它将立即将基础科学发现转化为切实的临床益处。 公共健康相关性：拟议的研究与公共健康相关，因为血管异常（VA）代表了一个重要的临床问题，但治疗选择很少。成功开发出一种小分子或 FDA 批准的针对突变 Dusp-5 而非 WT Dusp-5 蛋白的化合物将为 VA 患者提供急需的治疗替代方案。因此，拟议的研究与 NIH 的使命直接相关，即减轻因脉管系统失调影响的疾病造成的健康状况恶化的负担。