Rusalatide Acetate (TP508) Mitigation of Genotoxic Radiation Damage in Human Lens Epithelial Cells

醋酸鲁沙拉肽 (TP508) 减轻人晶状体上皮细胞的基因毒性辐射损伤

• 批准号: 10384634
• 负责人: USHA P ANDLEY
• 金额: $ 32.75万
• 依托单位: AFFIRMED PHARMA, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10384634
• 关键词: Acetates; Amyloid beta-Protein; Animal Model; Animals; Apoptosis; BCL2 gene; Biological Assay; Blindness; CASP3 gene; CHEK1 gene; CHEK2 gene; Cancer Patient; Cataract; Cell Cycle; Cell Cycle Regulation; Cell Death; Cell Line; Cell Proliferation; Cell Survival; Cells; Complex; Cornea; Crystallins; Cyclins; DNA Double Strand Break; DNA Repair; DNA-PKcs; Data; Disease; Dose; Double Strand Break Repair; Drug Delivery Systems; Endothelial Cells; Epithelial; Epithelial Cells; Event; Family; Functional disorder; Future; Genes; Health; Hour; Human; Individual; Investigation; Laboratories; Lead; Lens Fiber; Life; Medical Economics; Methods; Mitotic; Molecular; Necrosis; Occupational Exposure; Oral mucous membrane structure; Pathway interactions; Peptides; Persons; Pharmaceutical Preparations; Phase; Population; Procedures; Proteins; Proton Radiation; Radiation; Radiation Dose Unit; Radiation Induced DNA Damage; Radiation Protection; Radiation Tolerance; Radiation exposure; Radiation induced damage; Radiation therapy; Radio; Retina; Retinal Diseases; Risk; Roentgen Rays; Schedule; Signal Pathway; Signal Transduction Pathway; Signaling Protein; Social Impacts; Societies; Stains; TP53 gene; Techniques; Testing; Therapeutic; Time; Tissues; Treatment Protocols; Western Blotting; Xerostomia; dosage; economic impact; eye dryness; fiber cell; genotoxicity; health related quality of life; improved outcome; irradiation; lens; optimal treatments; p53-binding protein 1; prevent; promoter; protective effect; protein aggregation; protein expression; radiation adverse effect; regenerative; response; senescence; side effect; single fraction radiation

Cancer patients benefit from radiation therapy but can incur side effects to non-targeted tissues including cataracts. Although not directly life threatening, cataract disease has major medical, economic, and social impacts on individuals, families, and society as a whole. Radiation-induced lens opacification is a complex event and has been attributed to DNA double strand breaks (DSB) in the germinative epithelium, leading to defective differentiation of lens fiber cells and subsequent abnormal folding of lens proteins. Rusalatide acetate (TP508) is a radio-modulating peptide that has been shown to increase survival of irradiated animals via activation of signal transduction pathways in endothelial cells, initiating repair of DSB, increasing NO levels and reversing of endothelial cell dysfunction. This investigation will determine if, in the absence of endothelial cells, TP508 will have a similar effect on human lens epithelial cells (HLEC) and mitigate radiation induced pathophysiological pathways that lead to DSB. The hypothesis is that through the direct activation of molecular pathways in irradiated HLEC, TP508 treatment will mitigate or repair DSB. In contrast to other investigative approaches that focus on a single downstream mechanism, this investigation will examine molecular activity of TP508 across multiple pathophysiological pathways associated with the health of HLEC. Study aims are to establish the molecular activity and optimum dosage thresholds, and timing of treatments of TP508 in mitigating X-ray or proton radiation damage with single fraction exposures of 0.5, 1.0, 2.0, or 4 Gy in HLEC (CRL-11421 [B3] and SRA01/04 lines). Aim (1) is to determine the most optimized concentration and administration schedule for TP508 effects on radiation induced HLEC viability using a clonogenic survival assay, MTT assay, and cell doubling time to assess the effects of various concentrations of TP508 on the sensitivity of HLECs applied before and after irradiation. Aim (2) is to further identify the effects of TP508 on specific HLEC molecular responses, using the most optimized concentration and administration schedule of TP508 comparing the single fraction radiation exposures applied with or without TP508, and analyzed with assays for apoptosis, necrosis, senescence, mitotic catastrophe and protein expression; 53BP1 foci staining for dynamics of DSB; and western blot assays for related signaling pathways and protein profiles including amyloid beta, an early marker of cataract formation. Studies are expected to provide the following: (i) determine if TP508 provides a survival effect on irradiated HLEC at different doses applied before and after radiation; (ii) identify if the molecular mechanisms and protein profiles underlying the protective effect of TP508 in HLEC at different doses of radiation are attributed to DSB repair; and (iii) determine the most effective dosage and timing (before or after radiation) of TP508 application. Successfully developed, future investigations of TP508 could expand its application to mitigate additional radiation side effects including dry eye, retinopathy, and xerostomia. .

癌症患者受益于放射治疗，但可能会对非目标组织产生副作用，包括 白内障。白内障疾病虽然不会直接危及生命，但它对医疗、经济和社会影响重大。 对个人、家庭和整个社会的影响。辐射引起的晶状体混浊是一种复杂的 事件被归因于萌发上皮中的 DNA 双链断裂 (DSB)，导致 晶状体纤维细胞分化缺陷以及随后的晶状体蛋白折叠异常。鲁沙拉肽 醋酸盐 (TP508) 是一种放射调节肽，已被证明可以提高受辐射的存活率 通过激活动物内皮细胞中的信号转导途径，启动 DSB 修复，增加 NO 水平和内皮细胞功能障碍的逆转。这项调查将确定，在没有 内皮细胞，TP508将对人晶状体上皮细胞（HLEC）产生类似的作用并减轻 辐射诱发导致 DSB 的病理生理学途径。假设是通过直接 TP508 治疗可激活受辐射 HLEC 中的分子途径，从而减轻或修复 DSB。在 与其他专注于单一下游机制的调查方法相比，本次调查 将检查 TP508 跨多种病理生理学途径的分子活性，这些途径与 HLEC 的健康状况。研究目的是确定分子活性和最佳剂量阈值，以及 TP508 减轻单次暴露 X 射线或质子辐射损伤的治疗时机 HLEC 中的剂量为 0.5、1.0、2.0 或 4 Gy（CRL-11421 [B3] 和 SRA01/04 系）。目标 (1) 是确定最 TP508 对辐射诱导的 HLEC 活力影响的优化浓度和给药方案 使用克隆生存测定、MTT 测定和细胞倍增时间来评估各种因素的影响 TP508 浓度对照射前后应用的 HLEC 的敏感性的影响。目标 (2) 是 使用最优化的方法进一步确定 TP508 对特定 HLEC 分子反应的影响 TP508 的浓度和给药方案比较单次辐射暴露 使用或不使用 TP508 进行应用，并通过细胞凋亡、坏死、衰老、有丝分裂分析进行分析 灾难和蛋白质表达； 53BP1 焦点染色用于 DSB 动力学；和蛋白质印迹分析 相关信号通路和蛋白质谱，包括β淀粉样蛋白（白内障的早期标志物） 形成。研究预计将提供以下内容：(i) 确定 TP508 是否对患者具有生存作用 辐射前和辐射后以不同剂量照射的 HLEC； (ii) 确定分子是否 TP508 在不同剂量下对 HLEC 的保护作用的机制和蛋白质谱 辐射归因于 DSB 修复； (iii) 确定最有效的剂量和时间（之前或之后） TP508 应用的辐射后）。 TP508开发成功，未来研究范围可扩大 它的应用可减轻额外的辐射副作用，包括干眼症、视网膜病变和口干症。 。