(9) Therapeutic strategies to mitigate toxicities of platinum-based therapeutics

摘要 顺铂是应用最广泛的抗癌药物之一，包括用于治疗 无论是孩子还是成年人。顺铂的临床应用与对肾小管细胞的剂量限制损伤有关 (肾毒性)，尽管采取了密集的预防措施，但高达40%的患者发生了肾毒性，以及 对耳蜗(耳毒性)和周围神经(神经毒性)的毒性，这些并发症可能会进一步限制 治疗，甚至威胁生命。目前还没有已知的针对顺铂引起的毒性的特定治疗方法， 这些副作用的机械细节仍然知之甚少。我们最近发现，这种能力 铂类化疗药物对健康组织细胞的损伤依赖于有机阳离子 对于肾小管细胞，还有有机阴离子转运体(OAT)。在小鼠身上，这些 研究发现，这些过程受到两对密切相关的转运蛋白Oct1/Oct2的调控 顺铂的细胞摄取，以及调节顺铂硫代谢酸代谢物肾摄取的Oat1/OAT3 这是一种强肾毒素的前驱物质。我们发现这4种转运蛋白的功能可以是 被酪氨酸激酶抑制剂尼洛替尼通过非竞争性机制有效抑制。在当前 建议，我们概述了三组相关研究，将进一步测试和完善我们的中央 假设尼洛替尼靶向抑制OCT和OAT功能将特异性地影响 顺铂在健康靶组织中的分布及其下游毒性效应：(I)使用不同的体外和体内实验 模型，包括斑马鱼和小鼠，我们将进一步确定其定性和定量的影响 尼洛替尼对Oct1/Oct2和Oat1/Oat3功能的影响；(Ii)斑马鱼和小鼠的实验研究 感兴趣的转运蛋白和激酶将决定这些蛋白对顺铂相关的直接贡献。 毒性，包括肾毒性、耳毒性和神经毒性，以及药物处置特性和药物. 药物相互作用潜力；(Iii)使用有或没有来自复制细胞的人肿瘤异种移植的小鼠 我们将确定这些蛋白对顺铂相关抗癌作用的直接贡献。 尼洛替尼对这一表型的疗效和体内效应。顺铂诱导的减毒作用的实验研究 通过抑制调节药物进入健康组织的关键转运体的毒性将提供 为未来旨在改善该制剂的衰弱副作用的进一步研究奠定基础 常规的临床实践。

ABSTRACT Cisplatin is one of the most widely used anticancer agents, including for the treatment of solid tumors in children and adults. The clinical use of cisplatin is associated with dose-limiting damage to renal tubular cells (nephrotoxicity), which occurs in up to 40% of patients despite intensive prophylactic measures, as well as with toxicity to cochlea (ototoxicity) and peripheral nerves (neurotoxicity), and these complications may limit further treatment or even threaten life. There is currently no known specific treatment for cisplatin-induced toxicities, and mechanistic details of these side effects remain poorly understood. We have recently found that the ability of platinum chemotherapeutics to cause damage to healthy tissues cells is dependent on organic cation transporters (OCT) and, for renal tubular cells, organic anion transporters (OAT) as well. In mice, these processes were found to be regulated by the two pairs of closely related transporters, Oct1/Oct2 that regulate cellular uptake of cisplatin, and Oat1/Oat3 that regulate renal uptake of a cisplatin mercapturic acid metabolite that acts as a precursor of a potent nephrotoxin. We found that the function of these 4 transporters can be potently inhibited by the tyrosine kinase inhibitor nilotinib, through non-competitive mechanisms. In the current proposal, we outline three sets of related studies that will further test and refine the validity of our central hypothesis that targeted inhibition of OCT and OAT function with nilotinib will specifically affect accumulation of cisplatin in healthy target tissues and affect its downstream toxic effects: (i) Using various in vitro and in vivo models, including zebrafish and mice, we will further determine the qualitative and quantitative effects of nilotinib on the function of Oct1/Oct2 and Oat1/Oat3; (ii) Studies in zebrafish and mice with or without specific transporters and kinases of interest will determine the direct contribution of these proteins to cisplatin-related toxicity, including nephrotoxicity, ototoxicity, and neurotoxicity, as well as drug disposition properties and drug- drug interaction potentials; (iii) Using mice with or without human tumor xenografts derived from replicating cell lines or patient tumors, we will determine the direct contribution of these proteins to cisplatin-related anticancer efficacy, and the in vivo effects of nilotinib on this phenotype. The demonstration of reduced cisplatin-induced toxicity through inhibition of critical transporters regulating access of the drug to healthy tissues will provide the foundation for additional studies in the future aimed at ameliorating this agent's debilitating side effects in routine clinical practice.