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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要淋巴水肿是乳腺癌和妇科癌症放射和/或手术后的主要并发症。手术技术的进步降低了风险，但淋巴水肿的发生率仍然很高，并且存在没有批准的药物可以预防或治疗它。阿霉素 (DOX) 是一种用于治疗的主要化疗药物。乳腺癌和妇科癌症，但它会使淋巴水肿的风险增加 3 倍。 DOX 是否导致慢性淋巴水肿尚不清楚，但我们发现临床相关的 DOX 浓度通过激活兰尼碱受体（RYRs，淋巴肌细胞 (LMC) 中的细胞内钙通道），导致肌浆中的强效 Ca2+ 渗漏网状组织 (SR) 和淋巴组织持续高水平的胞质 Ca2+ [Ca2+i] 可促进脂质过氧化。和细胞死亡途径，以及淋巴淤积产生的腔内压力增加会损害左心室血管壁和瓣叶，协同导致慢性淋巴损伤目前尚不清楚 DOX 是否会激活。事实上，DOX 通过直接相互作用或通过介导 RYR 氧化间接提高两者。细胞质和线粒体超氧化物 (O2•-)，可能有助于 RYR 氧化（受体开放）和随后的 Ca2+ 泄漏还不清楚 DOX 激活哪种 RYR 亚型（RYR1、RYR2、RYR3）。 LMCs；如果已知，它可以作为预防 DOX 引起的淋巴功能障碍的潜在治疗靶点。我们认为 RYR 是 LMC 中的新治疗靶点，可预防 DOX 诱导的淋巴功能障碍和我们勇敢地面对DOX 产生O2•- 的剧烈氧化和开放。 RYR 增加 LMC 中的 [Ca2+i]，抑制左心室收缩并诱导淋巴淤滞和淋巴损伤，以及增加的手术损伤会增强这种效果，因此，我们将使用我们完善的大鼠模型来评估。 RYR 作为预防 DOX 引起的淋巴功能障碍的治疗靶点将整合技术。在临床前研究中探索这一假设，并将依赖于分离的 LV 的蛋白质和功能分析，使用光学成像来评估体内对 DOX 和 RYR 阻断的反应体积淋巴流量，以及研究 RYR 阻滞剂作为淋巴管临床前模型中潜在治疗方法的效用目标 1 将确定 LMC 中 DOX 诱导的 RYR 激活是否由 O2•- 介导。目标 3 将定义 RYR 亚型在 DOX 诱导的离体 LV 中 Ca2+ 渗漏和体内淋巴流动中的作用。研究 DOX ± RYR 阻滞剂对淋巴功能、脂质过氧化和淋巴管的综合影响因此，我们计划探索 RYR 作为新型淋巴管功能不全的临床前大鼠模型。预防 DOX 相关淋巴水肿的治疗靶点并评估 RYR 阻滞剂是否可用作抗淋巴水肿剂。