Drug development for tuberous sclerosis complex and other pediatric epileptogenic diseases using neurovascular and cardiac microphysiological models

使用神经血管和心脏微生理学模型开发结节性硬化症和其他儿科癫痫性疾病的药物

• 批准号: 10240589
• 负责人: KEVIN C ESS
• 金额: $ 114.92万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-21 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240589
• 关键词: Academic Medical Centers; Address; Affect; Aftercare; Animal Model; Astrocytes; Award; Biological Markers; Biological Models; Biomedical Engineering; Blood - brain barrier anatomy; Brain; Brain Neoplasms; Butadiene; Cardiac; Cardiac Myocytes; Cell Line; Cells; Childhood; Clinical Trials; Complex; Development; Disease; Disease model; Drug Targeting; Drug toxicity; Education; Electrophysiology (science); Endothelial Cells; Ensure; Epilepsy; Ethylenes; FDA approved; FRAP1 gene; Failure; Functional disorder; Future; Generations; Genetic Diseases; Goals; Health; Heart; Hematological Disease; Human; In Vitro; Institutes; Knowledge; Lead; Mass Spectrum Analysis; Measures; Methodology; Microfluidics; Modeling; Myocardial dysfunction; Neurologic; Neurological Models; Neuronal Dysfunction; Neurons; Outcome; Outcome Measure; Pathogenesis; Patients; Perfusion; Pericytes; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Phosphotransferases; Physiology; Research; Role; Safety; Seizures; Signal Pathway; Signal Transduction; Signaling Protein; Sirolimus; Styrenes; Technology; Testing; Tissue Microarray; Tissue Model; Toxic effect; Translations; Treatment Efficacy; Tuberous sclerosis protein complex; Validation; Vigabatrin; bench to bedside; biomarker development; biomarker identification; chromatin immunoprecipitation; design; drug candidate; drug development; drug efficacy; effective therapy; efficacy evaluation; heart function; high-throughput drug screening; human disease; human tissue; improved; individual patient; induced pluripotent stem cell; inhibitor/antagonist; ion mobility; mTORopathies; microphysiology system; model development; nervous system disorder; neurovascular; neurovascular unit; novel; novel therapeutics; organ on a chip; polydimethylsiloxane; pre-clinical; precision medicine; programs; protein biomarkers; protein metabolite; rare genetic disorder; relating to nervous system; response; specific biomarkers; therapy development; tool

The goal of this proposal is to establish in vitro tissue chip models of the closely related neurological disorders tuberous sclerosis complex (TSC) epilepsy, DEPDC5-associated epilepsy, and their associated cardiac dysfunction. The proposed research leverages emerging bioengineering technology for microphysiological systems developed at the Vanderbilt Institute for Integrative Biosystems Research and Education (VIIBRE) with human induced pluripotent stem cell tools in regular use at Vanderbilt University Medical Center to ask probing questions about genetic disorders that afflict the heart and brain and about the drugs to treat them. The VIIBRE neurovascular unit (NVU)/blood-brain barrier and cardiac I-Wire organ-on-chip models will test the hypothesis that mTORC1 and mTORC2 signaling differentially affect neural and cardiac dysfunction in TSC- and DEPDC5-associated epilepsy. The primary and shared abnormality in patients with TSC and DEPDC5- associated epilepsy is dysregulation of the mTOR kinase complex 1 (mTORC1) signaling pathway. TSC also has abnormalities in mTORC2 signaling not seen in DEPDC5-associated epilepsy. A focus on mTOR signaling in these human mTORopathies has several advantages. First, rapamycin and related compounds are FDA- approved mTORC1 inhibitors and have been shown to have efficacy in some aspects of the disease manifestations of TSC. Second, TSC- and DEPDC5-associated epilepsy are both associated with neural and cardiac dysfunction. Third, the role for compensatory or differential mTORC2 activity is unclear and controversial. For patients with TSC, drugs targeting the mTORC1 signaling pathway have been associated with shrinkage of brain tumors, reduced seizures, and improved cardiac function. Thus, drug development for this group of diseases is well suited for study using both the NVU and I-Wire cardiac-tissue chips. In its first two years, the project will develop the NVU and I-Wire disease models, aimed at refining the TSC and DEPDC5 NVU model; applying the I-Wire model to TSC and DEPDC5 cardiomyocytes; and validating outcome methodologies in control and patient-derived NVU and I-Wire chips. The next three years aim to evaluate, for biomarker identification in control, TSC, and DEPDC5 NVU and I-Wire chips, changes in mTORC1 and mTORC2 signaling, protein markers of cellular health and toxicity, metabolites, functional measures and electrophysiological activity; and, use ion mobility-mass spectrometry to evaluate NVU and I-Wire outcome measures plus drug metabolites after treatment with mTORC1 inhibitor rapamycin, the seizure drug vigabatrin, and novel pre-clinical mTOR drug candidates. The NVU and I-Wire will assess the efficacy and toxicity of these agents and define TSC/DEPDC5 shared vs disease-specific effects. With this organ-on-chip/human induced pluripotent stem cell platform, it will be possible to address currently confounding mechanisms of pathogenesis, identify new disease biomarkers, quantify how drugs cross the normal and diseased blood-brain barrier, and ultimately develop effective therapies and hence enable bench-to-bedside translation.

本研究的目的是建立与神经系统疾病密切相关的体外组织芯片模型 结节性硬化症（TSC）癫痫、DEPDC 5相关癫痫及其相关心脏 功能障碍拟议的研究利用新兴的生物工程技术， 范德比尔特综合生物系统研究与教育研究所（VIIBRE）开发的系统 与范德比尔特大学医学中心经常使用的人类诱导多能干细胞工具一起， 探索有关影响心脏和大脑的遗传疾病以及治疗这些疾病的药物的问题。 VIIBRE神经血管单元（NVU）/血脑屏障和心脏I-Wire器官芯片模型将测试 假设mTORC 1和mTORC 2信号传导差异性地影响TSC中神经和心脏功能障碍， 和DEPDC 5相关癫痫。TSC和DEPDC患者的原发性和共有异常5- 与癫痫相关的是mTOR激酶复合物1（mTORC 1）信号通路的失调。TSC还 在DEPDC 5相关癫痫中未观察到mTORC 2信号异常。关注mTOR信号传导 在这些人类mTOR病中具有几个优点。首先，雷帕霉素和相关化合物是FDA- 已被批准的mTORC 1抑制剂，并已被证明在疾病的某些方面具有疗效 TSC的表现。其次，TSC和DEPDC 5相关癫痫均与神经和 心功能不全第三，补偿性或差异性mTORC 2活性的作用尚不清楚， 争议对于TSC患者，靶向mTORC 1信号通路的药物与 脑肿瘤缩小，癫痫发作减少，心脏功能改善。因此，药物开发 这组疾病非常适合使用NVU和I-Wire心脏组织芯片进行研究。在其前两场 该项目将开发NVU和I-Wire疾病模型，旨在完善TSC和DEPDC 5 NVU模型;将I-Wire模型应用于TSC和DEPDC 5心肌细胞;并验证结果 控制和患者衍生NVU和I-Wire芯片的方法。未来三年的目标是评估， 对照、TSC和DEPDC 5 NVU和I-Wire芯片中的生物标志物鉴定，mTORC 1和 mTORC 2信号传导、细胞健康和毒性的蛋白质标志物、代谢物、功能测量和 电生理活性;以及，使用离子迁移率-质谱法评价NVU和I-Wire结局 在用mTORC 1抑制剂雷帕霉素，癫痫发作药物氨己烯酸， 和新的临床前mTOR候选药物。NVU和I-Wire将评估这些器械的疗效和毒性。 药物和定义TSC/DEPDC 5共享与疾病特异性效应。通过这种芯片上的器官/人类诱导 多能干细胞平台，将有可能解决目前混淆的机制， 发病机制，确定新的疾病生物标志物，量化药物如何穿过正常和患病的血脑 屏障，并最终开发出有效的治疗方法，从而实现从实验室到床边的转变。

项目成果

The Microbiome and the Gut-Liver-Brain Axis for Central Nervous System Clinical Pharmacology: Challenges in Specifying and Integrating In Vitro and In Silico Models.

• DOI: 10.1002/cpt.1870
• 发表时间: 2020-11
• 期刊: Clinical pharmacology and therapeutics
• 影响因子: 6.7
• 作者: Hawkins KG;Casolaro C;Brown JA;Edwards DA;Wikswo JP
• 通讯作者: Wikswo JP

Application of a Human Blood Brain Barrier Organ-on-a-Chip Model to Evaluate Small Molecule Effectiveness against Venezuelan Equine Encephalitis Virus.

• DOI: 10.3390/v14122799
• 发表时间: 2022-12-15
• 期刊: Viruses
• 作者: Boghdeh NA;Risner KH;Barrera MD;Britt CM;Schaffer DK;Alem F;Brown JA;Wikswo JP;Narayanan A
• 通讯作者: Narayanan A

GROWTH AND CHARACTERIZATION OF A TISSUE-ENGINEERED CONSTRUCT FROM HUMAN CORONARY ARTERY SMOOTH MUSCLE CELLS.

• DOI: 10.20538/1682-0363-2020-2-85-95
• 发表时间: 2020-07
• 期刊: Biulleten' Sibirskoi meditsiny
• 作者: A. A. Sulgin-A.;T. Sidorova;V. Sidorov