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.

项目摘要 生长马（GH）是一种单链多肽，是细胞生长，增殖和 哺乳动物的代谢。 GH采取行动促进儿童期间纵向生长和适当的器官发育 - 引擎盖并刺激肝脏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GH活性降低会导致健康衰老并增加 寿命;实际上，寿命最长的实验室小鼠是由于GH受体（GHR）基因的全球破坏而导致的。 值得注意的是，这种GHR - / - 小鼠已经表现出对GH介导的疾病的抗性增强 不健康的衰老，包括糖尿病最终器官损伤和某些类型的癌症。这之间的联系 降低的GH作用和延长的寿命导致了这样的假设，即抑制GH作用可能会延迟发作 与年龄有关的病毒。肽代表了一种有吸引力的分子，可以用作治疗铅 因为它们可以设计为模仿蛋白质相互作用域的可变结构和序列。 此外，胡椒体是特定于序列的，并且可以合成牵引，从而使它们绕过许多 与基于蛋白质的药物相关的生产问题。我们的小组最近确定了一种基于肽的新型 GHR拮抗剂（称为SH1），可减轻培养细胞系中的GH介导的信号传导。 S1H被设计为 与GHR相互作用的GH的小螺旋区域（残基36-51）的直接序列模仿。结构 S1H的活性关系表明肽的螺旋性与GHR拮抗作用之间有很强的相关性， 我们假设生物活性需要螺旋前景。我们现在寻求检验这个假设 通过开发折叠成稳定螺旋结构的S1H衍生物并使用它们抑制GH介导的 体外和体内信号传导。在本提案的目标1中，我们将合成两种单独的结构化S1H类 衍生物。一类将通过将烯烃的侧链安装到野生型S1H序列中来生成 可以将肽“钉”成A螺旋结构。第二类将通过转移S1H开发 残留在Scylatoxin的A螺旋上，一种小蛋白质，可折叠成稳定的A/B基序。产生的肽 然后将用于针对重组GHR的一系列直接结合和电泳迁移率转移测定。 在AIM 2中，我们将研究结构化S1H衍生物抑制细胞中GH信号传导的能力 过表达衰老的GHR和体内小鼠模型。首先，我们将采用基于细胞的替代测定法 确定结构化的S1H衍生物是否可以抑制GH介导的下游磷酸化 转录因子，例如STAT5。接下来，我们将评估结构化S1H衍生物如何影响血清水平 IGF-1和IGF-BP3，以及C57BL6小鼠的身体组成。该提案产生的数据将作为 探索S1H（及其结构化的分子机制）的未来研究基础 衍生物）影响GH介导的信号传导，并将识别潜在的铅化合物作为新兴的铅化合物 旨在减轻GH介导的疾病的治疗剂，导致不健康的衰老。