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

基本信息

批准号：
10704484
负责人：
Steven Jay Frank
金额：
$ 6.13万
依托单位：
AFFIRMED PHARMA, LLC
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10704484
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 Differentiation process Cell Line Cell Proliferation Cell Survival Cells Complex Crystalline Lens Crystallins Cyclins DNA Double Strand Break DNA Repair DNA-PKcs Data Disease Dose Double Strand Break Repair Drug Delivery Systems Drug toxicity Endothelial Cells Epithelial Epithelial Cells Event Family Functional disorder Future Hour Human Individual Investigation Laboratories Lead Lens Fiber Life Medical Economics Mitotic Modeling Molecular Necrosis Normal Cell Normal tissue morphology Occupational Exposure Pathway interactions Peptides Persons Pharmaceutical Preparations Phase Population Procedures Proteins Proton Radiation Protons Radiation Radiation Dose Unit Radiation Induced DNA Damage Radiation exposure Radiation induced damage Radiation therapy Radio Retinal Diseases Risk Roentgen Rays Safety Schedule Signal Pathway Signal Transduction Pathway Social Impacts Societies TP53 gene Techniques Testing Therapeutic Time Toxic effect Treatment Protocols Western Blotting dosage economic impact eye dryness fiber cell genotoxicity health related quality of life improved outcome lens misfolded protein molecular marker peptide drug promoter protective effect protein aggregation protein biomarkers protein folding protein misfolding radiation adverse effect regenerative response senescence side effect

项目摘要

Cancer patients benefit from radiation therapy but can incur side effects to normal tissues including the ocular lens, leading to 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 TP508 will have a similar effect on human lens epithelial cells (HLEC) and mitigate radiation-induced pathophysiological pathways that are associated with abnormal differentiation of fiber cells, abnormal protein folding and opacification. The hypothesis is that through the direct activation of molecular pathways in irradiated HLEC, TP508 treatment will mitigate or repair DSB and restore normal cell differentiation. 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 normal differentiation 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 damage with single fraction exposures of 0.5, 1.0, 2.0, or 4 Gy in HLEC (CRL-11421 [B3], SRA01/04 and HLEpiC cells. Aim (1) is to determine drug toxicity levels using clonogenic survival assays and cell doubling times to identify the optimum TP508 concentration and administration schedule (before or after radiation) for producing protective effects on radiation induced HLEC viability. Aim (2) will determine if TP508 can maintain or restore normal cell differentiation required for normal protein folding, using the most optimized dosage and administration schedule established in Aim 1. Investigations will include the effects of TP508 on molecular markers and proteins associated with abnormal differentiation of lens fiber cells (CRYAB [αB-crystallin], CRYBB2 [βB2-crystallin]) and proteins involved in signaling pathways for apoptosis, necrosis, senescence, and mitotic catastrophe. Studies are expected to provide the following: (i) determine limits of toxicity with increasing doses of TP508 and the survival effect on irradiated HLEC at different doses applied before and after radiation; and (ii) identify if the molecular mechanisms and protein markers associated with the progression of lens opacification are mitigated by the optimum doses of TP508 (from Aim 1) in HLEC at different levels of radiation. Successfully developed, future investigations of TP508 could expand its application to mitigate additional ocular radiation therapy side effects including dry eye and retinopathy.

癌症患者受益于放射治疗，但可能会对包括眼部在内的正常组织产生副作用晶状体，导致白内障，虽然不会直接危及生命，但白内障疾病具有重大的医疗、经济、对个人、家庭和整个社会的社会影响是辐射引起的晶状体混浊。复杂的事件，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晶状体蛋白的异常折叠。 Rusalatideacetate (TP508) 是一种放射调节肽，已被证明可以提高通过激活内皮细胞中的信号转导途径，启动 DSB 修复，增加 NO 水平并逆转内皮细胞功能障碍这项研究将确定 TP508 是否会增加。对人晶状体上皮细胞（HLEC）有类似的作用，并减轻辐射引起的与纤维细胞异常分化、异常蛋白质相关的病理生理途径折叠和不透明的假设是通过分子途径的直接激活。相比之下，经照射的HLEC、TP508治疗会减轻或修复DSB并恢复正常的细胞分化。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 跨多个与正常相关的病理生理学途径的分子活性研究目的是确定 HLEC 的分子活性和最佳剂量阈值，以及 TP508 在减轻 X 射线或质子损伤方面的治疗时机，单次暴露剂量为 0.5、1.0、 HLEC（CRL-11421 [B3]、SRA01/04 和 HLEpiC 细胞）中 2.0 或 4 Gy。目的 (1) 是确定药物毒性使用克隆存活测定和细胞倍增时间确定最佳 TP508 浓度以及对辐射产生保护作用的给药时间表（辐射前或辐射后） HLEC 活力将确定 TP508 是否能够维持或恢复正常细胞分化所需的能力。正常的蛋白质折叠，使用目标 1 中建立的最优化剂量和给药方案。研究将包括 TP508 对与异常相关的分子标记和蛋白质的影响晶状体纤维细胞（CRYAB [αB-晶状体蛋白]、CRYBB2 [βB2-晶状体蛋白]）和参与的蛋白质的分化细胞凋亡、坏死、衰老和有丝分裂灾难的信号通路研究预计将进行。提供以下内容：(i) 确定 TP508 剂量增加的毒性限度以及对生存的影响在辐射之前和之后以不同剂量照射的 HLEC；以及 (ii) 确定分子是否存在；与晶状体混浊进展相关的机制和蛋白质标记物通过不同辐射水平下 TP508（来自目标 1）在 HLEC 中的最佳剂量已成功开发，未来。 TP508 的研究可以扩大其应用，以减轻额外的眼部放射治疗副作用包括干眼症和视网膜病变。