Protective role of Neuregulin-1 against cerebral malaria-induced neuronal injury and behavioral sequelae

Neuregulin-1对脑型疟疾引起的神经元损伤和行为后遗症的保护作用

• 批准号: 10391193
• 负责人: BYRON D. FORD
• 金额: $ 55.78万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10391193
• 关键词: 3-Dimensional; ANGPT1 gene; Aftercare; Algorithms; Angiopoietin-2; Animal Model; Animals; Anti-Inflammatory Agents; Antibodies; Apoptosis; Apoptotic; Attenuated; Autopsy; Behavioral; Blood - brain barrier anatomy; Brain; Brain Edema; Brain Injuries; C57BL/6 Mouse; CD8-Positive T-Lymphocytes; CD8B1 gene; CXCL10 gene; Cell Culture Techniques; Cerebral Malaria; Cerebrum; Cessation of life; Child; Chimeric Proteins; Clinical Trials; Cognition; Collaborations; Coma; Conscious; Cytolysis; Dendritic Cells; Disease; Encephalitis; Epithelial Cells; ErbB4 gene; Erythrocytes; Ethics; Extracellular Matrix; Fatal Outcome; Functional disorder; Gene Expression; Genes; Goals; HRP-2 protein; Heart failure; Heme; Human; Human Experimentation; Impairment; In Vitro; Inbred BALB C Mice; Infection; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Integration Host Factors; Interferons; Intravenous; Magnetic Resonance Imaging; Malaria; Matrix Metalloproteinases; Mediating; Microcirculation; Modeling; Mus; NRG1 gene; Neuregulin 1; Neuronal Injury; Neuropeptides; Organoids; PD-1 blockade; PD-1/PD-L1; Parasites; Participant; Pathogenesis; Peptides; Peripheral Blood Lymphocyte; Phosphorylation; Plasmodium berghei; Plasmodium falciparum; Production; Proto-Oncogene Proteins c-akt; Recovery; Regulatory Pathway; Reporting; Resistance; Role; STAT3 gene; Serum; Severities; Severity of illness; Signal Pathway; Signal Transduction; Survival Rate; Survivors; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Therapeutic Agents; Tissues; Translating; Urine; artemether; attenuation; brain dysfunction; brain endothelial cell; brain tissue; cell injury; disease prognosis; effectiveness evaluation; genetic variant; heme oxygenase-1; human stem cells; human subject; improved; in vitro Model; induced pluripotent stem cell; monocyte; mortality; neglect; neurobehavioral; neurovascular unit; programmed cell death ligand 1; programmed cell death protein 1; prospective; receptor; recruit; tau Proteins

Human cerebral malaria (HCM) is a severe form of Plasmodium falciparum (P.f.) malaria associated with ~500,000 deaths in children annually and impaired brain function in some survivors. HCM is characterized by sequestration of parasitized red blood cells (pRBCs) in cerebral micro-circulation and induction of inflammatory mediators, brain swelling and impaired consciousness with unarousable coma. We have determined that circulating free heme and parasite histidine rich protein 2 (HRP2), by-products of pRBC lysis, are major causes of brain inflammation, blood-brain barrier (BBB) dysfunction, and brain injury associated with HCM. However, defining the mechanism(s) mediating these effects in HCM is challenging in the absence of suitable models. In vitro 2D cell culture, 3D brain organoid and animal models (ECM; Plasmodium berghei ANKA in C57BL/6) all indicate that heme and HRP2 induced cellular apoptosis, inflammation, and tissue disorganization. In ECM, heme causes brain vascular endothelial cell apoptosis, alters Angiopoietins 1 and 2 ratios, upregulates CXCL10, Heme Oxygenase 1, and tau as well as compromise BBB integrity through STAT3 signaling via matrix metalloproteinase three (MMP3). Following a screen for therapeutic agents against ECM, we identified Neuregulin- 1(NRG1), an 8 kDa neuropeptide currently undergoing clinical trials against heart failure, that attenuates ECM when delivered intravenously at 5µg/kg. NRG1 mediates phosphorylation of ErbB4 (receptor), activates AKT and inactivates STAT3 in human brain microvascular endothelial cells. ECM resistant mice (BALB/c) constitutively expressed higher levels of NRG1 in brain tissue than ECM susceptible (C57BL/6) mice. Since circulating NRG1 is severely depleted in both fatal HCM and ECM, levels of NRG1 need to be assessed prospectively to ascertain amounts needed for augmentation to mitigate HCM severity. Interestingly, CD8+Tcell PD1/PD-L1 signaling mediated ECM recovery and PD1 was upregulated by NRG1. Using a human stem cell- derived neurovascular unit (NVU; brain chip), ECM and human subjects, we will determine the mechanism by which NRG1 attenuates cerebral malaria. We hypothesize that therapeutic administration of NRG1 will attenuate heme and HRP2-induced NVU damage and ECM mortality via NRG1/ErbB4 and PD/PD-L1 signaling. Our objective is to functionally assess the key regulatory pathways mediated by NRG1 to attenuate ECM, and heme- and HRP2-induced NVU damage. The specific aims are: 1) to test the hypothesis that an algorithm, consisting of NRG1, heme, HRP2, and markers of neuronal injury and inflammation can predict HCM severity, mortality and neurobehavioral sequelae; 2) to test the hypothesis that NRG1/ErbB4 and PD1/PD-L1 signaling crosstalk protects against heme and HRP2-induced damage in human NVU; and 3) to test the hypothesis that NRG1 attenuates ECM brain injury and behavioral deficit via NRG1/ErbB4 and PD1/PD-L1 signaling. Understanding the role of NRG1 in cerebral malaria pathogenesis and sequelae in survivors will enable us to determine the feasibility of targeting NRG1 in clinical trials with the ultimate goal of improving the survival of children with HCM.

人脑型疟疾（HCM）是恶性疟原虫（P.f.）疟疾与 每年约有50万儿童死亡，一些幸存者的脑功能受损。HCM的特点是 脑微循环中寄生红细胞（pRBC）的隔离和炎症反应的诱导 介质，脑肿胀和意识障碍伴无法唤醒的昏迷。我们已经确定 循环游离血红素和寄生虫富组氨酸蛋白2（HRP 2），pRBC裂解的副产物，是主要原因 脑炎症、血脑屏障（BBB）功能障碍和与HCM相关的脑损伤。然而，在这方面， 在缺乏合适的模型的情况下，定义HCM中介导这些效应的机制是具有挑战性的。在 体外2D细胞培养、3D脑类器官和动物模型（ECM; C57 BL/6中的伯氏疟原虫ANKA）， 表明血红素和HRP 2诱导细胞凋亡、炎症和组织解体。在ECM中， 血红素引起脑血管内皮细胞凋亡，改变血管生成素1和2的比例，上调CXCL 10， 血红素加氧酶1和tau以及通过基质STAT 3信号传导损害BBB完整性 金属蛋白酶3（MMP 3）。在筛选针对ECM的治疗药物后，我们发现 Neuregulin- 1（NRG 1）是一种目前正在进行抗心力衰竭临床试验的8 kDa神经肽， 当以5μg/kg静脉内递送时，可减弱ECM。NRG 1介导ErbB 4（受体）的磷酸化， 在人脑微血管内皮细胞中激活AKT并灭活STAT 3。ECM抵抗小鼠 (BALB/c）在脑组织中组成性表达比ECM易感（C57 BL/6）小鼠更高水平的NRG 1。 由于循环NRG 1在致死性HCM和ECM中严重耗尽，因此需要评估NRG 1的水平。 前瞻性地确定缓解HCM严重程度所需的增加量。有趣的是，CD 8 + T细胞 PD 1/PD-L1信号传导介导ECM恢复，并且PD 1被NRG 1上调。使用人类干细胞- 衍生的神经血管单位（NVU;脑芯片），ECM和人类受试者，我们将确定的机制， NRG 1减弱脑型疟疾。我们假设NRG 1的治疗性给药将减弱 血红素和HRP 2通过NRG 1/ErbB 4和PD/PD-L1信号传导诱导NVU损伤和ECM死亡。我们 目的是从功能上评估NRG 1介导的减弱ECM和血红素的关键调节途径， 和HRP 2诱导的NVU损伤。具体的目的是：1）测试假设，一个算法，包括 NRG 1、血红素、HRP 2以及神经元损伤和炎症标志物的表达可以预测HCM的严重程度、死亡率、 2）验证NRG 1/ErbB 4和PD 1/PD-L1信号转导通路相互作用的假说， 保护免受血红素和HRP 2诱导的人NVU损伤;和3）测试NRG 1 通过NRG 1/ErbB 4和PD 1/PD-L1信号转导减轻ECM脑损伤和行为缺陷。理解 NRG 1在脑型疟疾发病机制和幸存者后遗症中的作用将使我们能够确定 在临床试验中靶向NRG 1的可行性，最终目标是改善HCM儿童的生存率。