Erythrocyte Autophagy Proteins as Potential Non-opioid Novel Targets for Pain in Sickle Cell Disease

项目摘要/摘要 这项提议的长期目标是开发非阿片类药物的新分子靶点，以减少 由镰状细胞病(SCD)引起的疼痛、痛苦和死亡，通过识别上游 急性和慢性疼痛信号的信使。SCD红细胞中线粒体滞留的研究 现在被认为在SCD的发病机制中起着重要作用。然而，下游 与疼痛和线粒体滞留相关的分子机制尚未被研究。我们的 初步数据表明，组织蛋白、GABARAPs的失调自噬副产物 而在SCD红细胞裂解过程中释放的突触核蛋白有可能引发疼痛 SCD患者的发病情况。在这里，我们假设特定的自噬蛋白包括 CTSO、CTSW和SNCB以及其他已知的DAMP信号分子由 SCD红细胞通过与疼痛信号分子相互作用在SCD中贡献疼痛 内皮和神经元。这一概念得到了初步数据的支持，这些数据表明SCD红细胞 随着自噬蛋白水平的增加，不仅在SCD红细胞中，在血浆中也是如此 到控制程序。为了解决这一假设，本文提出了两个目标。第一个目标将决定 SCD的疼痛程度是否与自噬的表达和活性水平有关 儿童和成人SCD患者中的蛋白质。第二个目标是确定是否 抑制自噬蛋白可以减少SCD小鼠的疼痛信号和疼痛行为 自噬蛋白突触核蛋白(SNCA， SNCB)、组织蛋白(CTSB、CTSO.CTSW)和具有疼痛信号的GABARAP(L1和L2) 分子。预计这些研究的结果将使人们了解 SCD中疼痛的机制，并最终将导致临床试验评估新的药理学 减少阿片类药物止痛需求的干预措施。

PROJECT SUMMARY/ABSTRACT The long-term objective of this proposal is to develop non-opioid novel molecular targets to reduce the pain suffering and mortality attributed to sickle cell disease (SCD), by identifying to upstream messengers of acute and chronic pain signaling. Mitochondrial retention in SCD red blood cells is recognized now to be a major contribution in SCD pathogenesis. However, the downstream molecular mechanisms related to pain and mitochondrial retention is not investigated. Our preliminary data suggested that dysregulated autophagy byproducts of Cathepsins, GABARAPs and synucleins released during lysis of RBCs of SCD red blood cells have potential to trigger pain episodes in SCD patients. Here, we hypothesized that specific autophagy proteins including CTSO, CTSW and SNCB along with other known DAMP signaling molecules generated from SCD RBCs contribute pain in SCD through interaction with pain signaling molecules in endothelium and neurons. This notion is supported by preliminary data indicating that SCD RBCs with increased levels of autophagy proteins not only in SCD RBCs but also in plasma compared to controls. To address this hypothesis, two aims are proposed. The first aim will determine whether the pain severity of SCD is associated with expression and activity levels of autophagy proteins in both pediatric and adults SCD patients. The second aim is to determine whether inhibiting autophagy proteins can reduce pain signaling and pain behaviors in an SCD mouse model and characterize the molecular interaction of autophagy proteins Synucleins (SNCA, SNCB), Cathepsins (CTSB, CTSO. CTSW) and GABARAPs (L1 and L2) with pain signaling molecules. It is anticipated that the result of these studies will make understand the unique mechanisms of pain in SCD and will ultimately lead to clinical trials to assess new pharmacological interventions to reduce the need for opiates to manage pain.