Elucidating anti-angiogenic tyrosine kinase inhibitor-induced vascular dysfunction

阐明抗血管生成酪氨酸激酶抑制剂诱导的血管功能障碍

基本信息

批准号：
10570393
负责人：
Mengcheng Shen
金额：
$ 12.85万
依托单位：
STANFORD UNIVERSITY
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：
2023-05-05 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570393
关键词：
3-Dimensional Address Advisory Committees Agonist Angiogenesis Inhibitors Animal Model Apoptosis Basic Science Biological Markers Biological Models Biology Biomedical Engineering Blood Vessels CRISPR interference CRISPR-mediated transcriptional activation CRISPR/Cas technology CXCL10 gene Cancer Patient Cancer Survivor Candidate Disease Gene Cardiac Cardiovascular Diseases Cardiovascular system Cell Communication Cell Death Induction Cell Survival Clinic Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Coronary Databases Development Disease Disease model Endothelial Cells Endothelium FDA approved FLT4 gene Foundations Functional disorder Genes Goals Harvest Heart Diseases Heart failure Human Individual Inflammation K-Series Research Career Programs Knowledge Lead Libraries Life Malignant Neoplasms Measures Mentors Mentorship Molecular Myocardial dysfunction Oncology Outcome Oxidative Stress Pathogenicity Patients Pericytes Permeability Pharmaceutical Preparations Phase Phenotype Physiological Plasma Pluripotent Stem Cells Positioning Attribute Predisposition Proteins Proteome Puromycin Receptor Protein-Tyrosine Kinases Reporting Research Research Personnel STAT3 gene Structure System TP53 gene Testing Time Tissue Model Tissues Toxic effect Training Translational Research Triage Trypsin Inhibitors Tyrosine Kinase Inhibitor Validation Vascular Diseases Whole Blood antagonist blood perfusion cancer therapy career career development cell type clinical translation clinically relevant comparison group cytotoxicity differential expression drug discovery druggable target efficacy evaluation efficacy validation endothelial stem cell engineered stem cells experience high-throughput drug screening human disease improved induced pluripotent stem cell inhibitor innovation insight knock-down mouse model multidisciplinary novel novel therapeutics overexpression personalized cancer therapy personalized therapeutic prevent research and development skills stem cell biology stem cell technology therapeutic candidate thrombogenesis transcriptome tumor vascular tissue engineering virtual

项目摘要

PROJECT SUMMARY Although tyrosine kinase inhibitors (TKIs) are highly potent in treating malignancies, >60% of them have been reported to lead to adverse cardiovascular outcomes. Moreover, anti-angiogenic TKIs, such as sunitinib, preferentially induce microvascular toxicity followed by cardiac dysfunction. However, our knowledge of the underlying pathogenic mechanisms of TKI-induced vascular toxicity (TKI-VT) has been hampered partially by the limited access to human diseased vascular tissues for molecular and cellular analysis. As such, no effective strategies have been developed to prevent or treat these otherwise life-threatening cardiovascular complications. This proposed project will leverage patient-specific induced pluripotent stem cell (iPSC)-derived cardiac pericytes (PCs) and endothelial cells (ECs) to understand the molecular and cellular basis of TKI-VT and discover personalized therapies for cancer patients who are suffering TKI-induced cardiovascular disease. In Aim 1, both monoculture and vessel-on-chip (VoC) coculture systems will be employed to characterize sunitinib-induced cell type-specific cytotoxicity profiles and aberrant cellular crosstalk between iPSC-PCs and iPSC-ECs that can contribute to TKI-VT. Overlapping upregulated and downregulated differentially expressed genes (DEGs) triggered by sunitinib in both cell types will serve as the candidate genes for large-scale druggable target screens. In Aim 2, CRISPR interference/activation (CRISPRi/a) survival screens will be performed in sunitinib-treated iPSC-PCs using a customized lentiviral sgRNA library targeting the overlapping DEGs identified in Aim 1. Top 10 hit genes in CRISPRi and CRSIPRa machineries will be subjected to structure-based virtual screens (SBVS) to discover candidate compounds that can mitigate TKI-VT phenotypes in iPSC-VoCs. In Aim 3, 3D iPSC- engineered vascular tissues (EVTs) and a mouse model will be used to validate the mitigation efficacy of candidate compounds on TKI-VT in a more physiological setting. The ex vivo plasma proteome generated from human whole blood-perfused 3D iPSC-EVTs will be correlated with those identified in patients to discover reliable disease-relevant biomarkers in predicting individual cancer patients’ susceptibility to TKI-VT. The research and career development training plans during the K99 phase, under the mentorship of Drs. Wu and Ky, as well as an expert interdisciplinary advisory committee, will provide Dr. Shen with advanced knowledge in stem cell biology, vascular biology, cardio-oncology, CRISPR technology, and bioengineering. The development of CRISPRi/a and SBVS screen platforms (K99) and 3D iPSC-EVTs (R00) will enable him to conduct disease modeling and drug discovery research in cardio-oncology specifically and vascular disease in general. The new skills and experience gained during this K99/R00 career development award, combined with Dr. Shen’s prior expertise in vascular biology, will facilitate his transition to an independent career conducting basic and translational research in cardio-oncology with a particular focus on the vascular aspect.

项目概要尽管酪氨酸激酶抑制剂 (TKI) 在治疗恶性肿瘤方面非常有效，但其中超过 60% 的药物已被用于治疗恶性肿瘤。据报道，抗血管生成 TKI，如舒尼替尼，会导致不良的心血管结局。优先引起微血管毒性，然后引起心脏功能障碍。 TKI 诱导的血管毒性 (TKI-VT) 的潜在致病机制部分受到以下因素的阻碍：对人类患病血管组织进行分子和细胞分析的途径有限，因此没有有效的方法。已经制定了预防或治疗这些危及生命的心血管并发症的策略。该拟议项目将利用患者特异性诱导多能干细胞 (iPSC) 衍生的心脏周细胞 (PC) 和内皮细胞 (EC) 了解 TKI-VT 的分子和细胞基础并发现在目标 1 中，针对患有 TKI 诱发的心血管疾病的癌症患者进行个性化治疗。将采用单一培养和芯片上血管 (VoC) 共培养系统来表征舒尼替尼诱导的细胞 iPSC-PC 和 iPSC-EC 之间的类型特异性细胞毒性特征和异常细胞串扰可以有助于 TKI-VT 的重叠上调和下调差异表达基因 (DEG)。舒尼替尼在两种细胞类型中触发的基因将作为大规模可药物靶点筛选的候选基因。在目标 2 中，将在接受舒尼替尼治疗的患者中进行 CRISPR 干扰/激活 (CRISPRi/a) 生存筛选 iPSC-PC 使用定制的慢病毒 sgRNA 文库，针对目标 1 中确定的重叠 DEG。 CRISPRi 和 CRSIPRa 机器中的 10 个命中基因将接受基于结构的虚拟筛选 (SBVS) 发现可以减轻 iPSC-VoC 中 TKI-VT 表型的候选化合物在目标 3 中，3D iPSC-。工程血管组织（EVT）和小鼠模型将用于验证缓解效果在更生理的环境中 TKI-VT 上的候选化合物产生的离体血浆蛋白质组。人类全血灌注的 3D iPSC-EVT 将与患者体内鉴定的相关，以发现可靠的预测个体癌症患者对 TKI-VT 易感性的疾病相关生物标志物。在 Wu 和 Ky 博士以及 Dr. 的指导下，制定了 K99 阶段的职业发展培训计划。跨学科专家顾问委员会将为沉博士提供干细胞方面的先进知识生物学、血管生物学、心脏肿瘤学、CRISPR技术和生物工程的发展。 CRISPRi/a 和 SBVS 筛选平台 (K99) 以及 3D iPSC-EVT (R00) 将使他能够进行疾病治疗特别是心脏肿瘤学和一般血管疾病的建模和药物发现研究。沉博士在获得 K99/R00 职业发展奖期间获得的技能和经验，以及之前的经验血管生物学方面的专业知识将有助于他过渡到独立的职业生涯，进行基础和心脏肿瘤学的转化研究，特别关注血管方面。