Developing helical peptide antagonists of the growth hormone receptor

开发生长激素受体的螺旋肽拮抗剂

• 批准号: 10648820
• 负责人: Justin Matthew Holub
• 金额: $ 41.53万
• 依托单位: OHIO UNIVERSITY ATHENS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648820
• 关键词: Acromegaly; Affect; Age of Onset; Aging; Amino Acid Sequence; Animal Model; Anterior Pituitary Gland; Binding; Biochemical; Biological; Biological Assay; Body Composition; Cell Line; Cell model; Cells; Childhood; Clinical; Cultured Cells; Data; Derivation procedure; Development; Diabetes Mellitus; Disease; Disulfide Linkage; Electrophoretic Mobility Shift Assay; FDA approved; Fluorescence Polarization; Foundations; Future; Goals; Growth; Growth Hormone Receptor; Heterogeneity; IGFBP3 gene; In Vitro; Injections; Insulin; Insulin-Like Growth Factor I; Laboratories; Laboratory mice; Lead; Life; Liver; Longevity; Malignant Neoplasms; Mammals; Marketing; Mediating; Metabolism; Molecular; Morbidity - disease rate; Mus; Nodular glomerulosclerosis; Organ; Patients; Peptides; Phenotype; Phosphorylation; Production; Proliferating; Proteins; Receptor Gene; Recombinant Growth Hormone; Recombinant Proteins; Research Personnel; Research Project Grants; Resistance; Series; Serum; Side; Signal Transduction; Somatotropin; Stat5 protein; Structure; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Intervention; Tissues; Variant; Vertebrates; Work; age related; alpha helix; antagonist; cancer type; cell growth; clinical application; cost; design; diabetic; experimental study; gel mobility shift assay; healthy aging; hormonal signals; immunogenicity; improved; in vivo; interest; mimetics; mouse model; novel; novel therapeutics; organ growth; overexpression; pegvisomant; peptide hormone; pleiotropism; polypeptide; postnatal; proliferative diabetic retinopathy; protein aminoacid sequence; protein folding; protein structure; reproductive; therapeutic protein; transcription factor; translational therapeutics

PROJECT SUMMARY Growth hormone (GH) is a single-chain polypeptide that acts as a key stimulator of cell growth, proliferation and metabolism in mammals. GH acts to promote longitudinal growth and proper organ development during child- hood and stimulates production of insulin-like growth factor 1 (IGF-1) in the liver and other tissues throughout life. Numerous studies have indicated that reduced GH activity in vivo results in healthy aging and increased lifespan; in fact, the longest-lived laboratory mouse results from global disruption of the GH receptor (GHR) gene. Notably, such GHR-/- mice have demonstrated enhanced resistance to GH-mediated disorders that contribute to unhealthy aging, including diabetic end organ damage and certain types of cancer. This connection between reduced GH action and extended lifespan has led to the hypothesis that inhibiting GH action may delay the onset of age-related morbidities. Peptides represent an attractive class of molecule to serve as therapeutic leads because they can be designed to mimic the variable structures and sequences of protein interaction domains. Moreover, peptides are sequence specific and synthetically tractable, allowing them to circumvent many of the production problems associated with protein-based drugs. Our group recently identified a novel peptide-based GHR antagonist (termed SH1) that mitigates GH-mediated signaling in cultured cell lines. S1H was designed as a direct sequence mimic of a small helical region (residues 36-51) of GH that interacts with the GHR. Structure activity relationships of S1H showed a strong correlation between peptide helicity and GHR antagonism, leading us to hypothesize that helical propensity is required for biological activity. We now seek to test this hypothesis by developing S1H derivatives that fold into stable helical structures and use them to inhibit GH-mediated signaling in vitro and in vivo. In aim 1 of this proposal, we will synthesize two separate classes of structured S1H derivative. The first class will be generated by installing olefinic side chains into the wild-type S1H sequence so the peptides can be ‘stapled’ into a-helical structures. The second class will be developed by transposing S1H residues onto the a-helix of scyllatoxin, a small protein that folds into a stable a/b motif. The resulting peptides will then be used in a series of direct binding and electrophoretic mobility shift assays against recombinant GHR. In aim 2, we will investigate the ability of our structured S1H derivatives to inhibit GH signaling in cells that overexpress the GHR and in vivo mouse models of aging. First, we will employ a cell-based surrogate assay to determine whether structured S1H derivatives can inhibit GH-mediated phosphorylation of downstream transcription factors, such as STAT5. Next, we will assess how structured S1H derivatives affect the serum levels of IGF-1 and IGF-BP3, and body composition of C57BL6 mice. Data generated from this proposal will serve as a foundation for future studies that explore the molecular mechanisms through which S1H (and its structured derivatives) affects GH-mediated signaling and will identify potential lead compounds to be used as emerging therapeutics designed to mitigate GH-mediated disorders that contribute to unhealthy aging.

项目总结 生长激素(GH)是一种单链多肽，是细胞生长、增殖和生长的关键刺激因子。 哺乳动物的新陈代谢。生长激素在儿童时期促进纵向生长和适当的器官发育 促进肝脏和其他组织中胰岛素样生长因子-1(IGF-1)的产生 生活。大量研究表明，体内生长激素活性降低会导致健康衰老和增加 寿命；事实上，最长寿的实验室小鼠是由生长激素受体(GHR)基因的全球破坏造成的。 值得注意的是，这样的GHR-/-小鼠对GH介导的疾病表现出了更强的抵抗力 到不健康的衰老，包括糖尿病末梢器官损害和某些类型的癌症。这种联系之间的联系 生长激素作用减弱和寿命延长导致了一种假设，即抑制生长激素作用可能会推迟发病 与年龄相关的疾病。多肽代表了一类具有吸引力的分子，可用作治疗先导。 因为它们可以被设计成模仿蛋白质相互作用结构域的可变结构和序列。 此外，多肽是特定于序列的，并且易于合成，使它们能够绕过许多 与蛋白质类药物相关的生产问题。我们的团队最近发现了一种新的基于多肽的 GHR拮抗剂(称为SH1)，可减轻培养细胞系中GH介导的信号转导。S1H设计为 与GHR相互作用的GH的一个小螺旋区域(残基36-51)的直接序列。结构 S1H的活性关系表明，多肽螺旋性与GHR拮抗作用有很强的相关性， 我们假设生物活动需要螺旋倾向。我们现在试图验证这一假设。 通过开发可折叠成稳定螺旋结构的S1H衍生物，并使用它们来抑制GH介导的 在体外和体内发出信号。在本提案的目标1中，我们将合成两类不同的结构化S1H 衍生品。第一类将通过将烯烃侧链安装到野生型S1H序列中而产生 这些多肽可以‘装订’成a-螺旋结构。第二类将通过转置S1H来开发 Syllatoxin的a-螺旋上的残基，这是一种折叠成稳定的a/b基序的小蛋白质。由此产生的多肽 然后将用于一系列针对重组GHR的直接结合和凝胶迁移率变化分析。 在目标2中，我们将研究我们的结构化S1H衍生物抑制细胞中生长激素信号转导的能力 过表达GHR和体内衰老的小鼠模型。首先，我们将使用基于细胞的代理检测来 确定结构性S1H衍生物是否能抑制GH介导的下游磷酸化 转录因子，如STAT5。接下来，我们将评估结构性S1H衍生物如何影响血清水平 对IGF-1、IGF-BP3和C57BL6小鼠体成分的影响。根据该提案生成的数据将用作 为未来探索S1H(及其结构)分子机制的研究奠定了基础 衍生物)影响GH介导的信号转导，并将确定潜在的先导化合物作为新兴的 旨在减轻生长激素介导的导致不健康衰老的疾病的治疗方法。