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. .

癌症患者受到放射疗法的受益 白内障。尽管没有直接威胁生命，但白内障疾病具有主要的医疗，经济和社会 对个人，家庭和社会的影响。辐射诱导的镜头无情是一个复合物 事件，并归因于发芽上皮中的DNA双链断裂（DSB），导致 透镜纤维细胞的有缺陷分化以及随后的透镜蛋白异常折叠。鲁萨拉丁 在 动物通过激活内皮细胞中信号转导途径的激活，启动DSB的修复，增加 没有水平和内皮细胞功能障碍的逆转。这项调查将确定是否没有 内皮细胞TP508将对人透镜上皮细胞（HLEC）具有相似的影响并减轻 辐射诱导导致DSB的病理生理途径。假设是通过直接 辐射的HLEC，TP508处理中的分子途径的激活将减轻或修复DSB。在 与其他关注单个下游机制的调查方法形成对比，该调查 将检查TP508的分子活性， HLEC的健康。研究目的是建立分子活性和最佳剂量阈值，以及 TP508治疗在减轻X射线或质子辐射损伤中的处理时间，单位分数暴露 HLEC中的0.5、1.0、2.0或4 Gy（CRL-11421 [B3]和SRA01/04线）。目的（1）是确定最多 TP508对辐射引起的HLEC生存能力的优化浓度和给药时间表 使用克隆生存测定，MTT分析和细胞加倍时间来评估各种影响 TP508在辐照前后应用HLEC的灵敏度上的浓度。目标（2）是 进一步确定TP508对特定HLEC分子响应的影响 TP508的浓度和给药时间表比较单分数辐射暴露 使用或不使用TP508的应用，并分析凋亡，坏死，衰老，有丝分裂的测定法 灾难和蛋白质表达； DSB动力学的53BP1焦点染色；和Western blot测定法 相关信号通路和蛋白质曲线包括淀粉样蛋白β，淀粉样蛋白β，白内障的早期标记 形成。期望研究提供以下内容：（i）确定TP508是否对 在辐射之前和之后，以不同剂量的不同剂量进行了辐照； （ii）确定是否分子 TP508在不同剂量的HLEC中的保护作用的基础的机理和蛋白质剖面 辐射归因于DSB修复； （iii）确定最有效的剂量和时机（或 辐射后TP508应用。成功开发的，对TP508的未来调查可能会扩大 它用于减轻其他辐射副作用，包括干眼症，视网膜病和静脉疾病。 。