Ryanodine Receptors as Therapeutic Targets to Prevent Doxorubicin-Induced Lymphatic Dysfunction

瑞尼定受体作为预防阿霉素引起的淋巴功能障碍的治疗靶点

• 批准号: 10712392
• 负责人: Amanda Stolarz
• 金额: $ 35.2万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712392
• 关键词: 3-Dimensional; Acute; Attenuated; Breast Cancer Patient; Breast Cancer therapy; Calcium Channel; Cancer Patient; Cell Death; Cell Death Induction; Chronic; Complication; Development; Doxorubicin; Drug usage; Enzymes; Generations; Healthcare Systems; Histologic; Immobilization; Impairment; Incidence; Injury; Ligation; Lipid Peroxidation; Liquid substance; Lymph; Lymphatic; Lymphatic function; Lymphedema; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Mediating; Mesentery; Mitochondria; Modeling; Monitor; Morphology; Muscle Cells; Operative Surgical Procedures; Pathway interactions; Patients; Perfusion; Periodicity; Persons; Pharmaceutical Preparations; Pre-Clinical Model; Proteins; Radiation; Radiation Dose Unit; Rattus; Receptor Activation; Research; Risk; Role; RyR1; RyR2; RyR3; Ryanodine Receptor Calcium Release Channel; SOD2 gene; Sarcoplasmic Reticulum; Second Primary Cancers; Signal Transduction; Speed; Sulfhydryl Compounds; Superoxides; Surgical Models; Techniques; Therapeutic; Tissues; breast surgery; cancer risk; cancer type; chemotherapy; clinically relevant; copper zinc superoxide dismutase; in vivo; interstitial; knock-down; lipidomics; lymph flow; lymph nodes; lymph stasis; lymphatic circulation; lymphatic dysfunction; lymphatic insufficiency; lymphatic surgery; lymphatic vessel; malignant breast neoplasm; new therapeutic target; novel; optical imaging; overexpression; oxidation; pre-clinical; preclinical study; preservation; pressure; prevent; receptor; recurrent infection; response; risk mitigation; small hairpin RNA; therapeutic target

PROJECT SUMMARY/ABSTRACT Lymphedema is a major complication after radiation and/or surgery for breast and gynecological cancers. Advancements in surgical techniques mitigate the risk, but the incidence of lymphedema is still high, and there are no approved medications to prevent or treat it. Doxorubicin (DOX) is a central chemotherapy drug for treating breast and gynecological cancers, but it increases the risk of lymphedema by 3-fold. The mechanism by which DOX contributes to chronic lymphedema is unknown, but we found that clinically relevant concentrations of DOX acutely inhibit lymph vessel (LV) contractions and reduce lymph flow by activating ryanodine receptors (RYRs, intracellular calcium channels) in lymph muscle cells (LMCs), resulting in tonic Ca2+ leak from the sarcoplasmic reticulum (SR) and lymphostasis. Sustained high levels of cytosolic Ca2+ [Ca2+i] can promote lipid peroxidation and cell death pathways, and the increase in intraluminal pressure produced by lymphostasis can damage LV walls and valve leaflets, synergistically causing chronic lymphatic injury. It is unclear whether DOX activates RYRs through a direct interaction or indirectly by mediating the oxidation of RYRs. Indeed, DOX elevates both cytosolic and mitochondrial superoxide (O2•-), which could contribute to RYR oxidation (receptor opening) and subsequent Ca2+ leak. It is also unknown which RYR subtype (RYR1, RYR2, RYR3) is activated by DOX in LMCs; if known, it could serve as a potential therapeutic target to prevent DOX-induced lymphatic dysfunction. We propose RYRs are novel therapeutic targets in LMCs to prevent DOX-induced lymphatic dysfunction and the development of chronic lymphedema. We hypothesize that DOX generates O2•- to acutely oxidize and open RYRs to increase [Ca2+i] in LMCs, inhibiting LV contractions and inducing lymphostasis and lymphatic injury, and added surgical insult potentiates this effect. Accordingly, we will use our well-established rat model to evaluate RYRs as therapeutic targets to prevent DOX-induced lymphatic dysfunction. Three aims will integrate techniques in preclinical studies to explore this hypothesis and will rely on protein and functional analysis of isolated LVs, use optical imaging to assess volumetric lymph flow in vivo in response to DOX and RYR blockade, and investigate the utility of RYR blockers, as a potential therapeutics in a preclinical model of lymphatic insufficiency. Aim 1 will determine whether DOX-induced RYR activation is mediated by O2•- in LMCs. Aim 2 will define the role of RYR subtypes in DOX-induced Ca2+ leak in isolated LVs and in lymph flow in vivo. Aim 3 will investigate the combined impact of DOX ± RYR blocker on lymphatic function, lipid peroxidation, and lymphatic morphology in a preclinical rat model of lymphatic insufficiency. Thus, we plan to explore RYRs as novel therapeutic targets to prevent DOX-related lymphedema and evaluate whether RYR blockers can be utilized as anti-lymphedema agents.

项目摘要/摘要 淋巴水肿是辐射和/或乳腺癌和妇科癌症手术后的主要并发症。 手术技术的进步可以减轻风险，但淋巴水肿的事件仍然很高，那里 没有批准的药物来预防或治疗它。阿霉素（DOX）是一种用于治疗的中央化学疗法药物 乳腺癌和妇科癌症，但它增加了淋巴水肿的风险3倍。该机制 DOX有助于慢性淋巴水肿，但我们发现临床上相关的DOX浓度 急性抑制淋巴血管（LV）收缩，并通过激活Ryanodine受体（Ryrs，Ryrs， 淋巴肌肉细胞（LMC）中的细胞内钙通道），导致肌浆​​中的补品Ca2+泄漏 网状（SR）和淋巴结。持续高水平的胞质Ca2+ [Ca2+ i]可以促进脂质过氧化 和细胞死亡途径，以及淋巴结炎产生的腔内压力的增加会损害LV 墙壁和瓣膜小叶，协同造成慢性淋巴损伤。目前尚不清楚DOX是否激活 通过直接相互作用或间接介导Ryrs的氧化。确实，Dox提升了这两个 胞质和线粒体超氧化物（O2• - ），这可能有助于RYR氧化（受体开口），并且 随后的CA2+泄漏。尚不清楚哪种RYR Subtype（RyR1，RyR2，RyR3）在DOX中激活 LMC;如果已知，它可以作为防止DOX诱导的淋巴功能障碍的潜在治疗靶标。 我们建议RYRS是LMC中新型治疗靶标，以防止DOX诱导的淋巴功能障碍和 慢性淋巴水肿的发展。我们假设DOX会生成O2• - 急性氧化物并打开 在LMC中增加[CA2+I]的Ryrs，抑制LV收缩，诱导淋巴结和淋巴损伤，以及 增加了手术损伤潜力。根据所有人，我们将使用我们公认的大鼠模型来评估 Ryrs作为治疗靶标，以防止DOX诱导的淋巴功能障碍。三个目标将整合技术 在临床前研究中，以探讨这一假设，并将依赖于蛋白质和分离的LVS的功能分析， 使用光学成像来评估体内体内的体积淋巴流，以应对DOX和RYR封锁，并且 研究RYR阻滞剂的效用，作为淋巴临床前模型中的潜在疗法 不足。 AIM 1将确定DOX诱导的RYR激活是否由LMC中的O2• - 。 AIM 2意志 定义了RYR亚型在DOX诱导的Ca2+泄漏中的作用，在体内和体内淋巴流中的作用。目标3意志 研究DOX±RYR阻滞剂对淋巴功能，脂质过氧化和淋巴的综合影响 淋巴功能不全的临床前大鼠模型中的形态。那就是我们计划探索里尔作为新颖的 预防与DOX相关淋巴水肿的治疗靶标，并评估RYR阻滞剂是否可以用作 抗淋巴水肿药。