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

• 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
• 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

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.

项目摘要 尽管酪氨酸激酶抑制剂（TKI）在治疗恶性肿瘤方面具有很高的潜力 据报道导致心血管不良后果。此外，抗血管生成的TKI，例如舒尼替尼， 优先诱导微血管毒性，然后引起心脏功能障碍。但是，我们对 TKI诱导的血管毒性（TKI-VT）的潜在致病机制已被部分阻碍 有限的访问人类的血管组织进行分子和细胞分析。因此，无效 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。 该提出的项目将利用特定于患者的诱导多能干细胞（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 针对患有TKI诱导心血管疾病的癌症患者的个性化疗法。在AIM 1中，两者都 将雇用单培养和片上的血管（VOC）共培养系统来表征Sunitinib诱导的细胞 IPSC-PC和IPSC-EC之间的类型特异性细胞毒性曲线和异常的细胞串扰可以 有助于TKI-VT。重叠的上调和下调不同表达的基因（DEGS） 两种细胞类型中由舒尼尼触发的触发，将作为大规模可药物目标筛查的候选基因。 在AIM 2中，CRISPR干扰/激活（CRISPRI/A）将在Sunitinib治疗中执行生存屏幕 IPSC-PC使用定制的慢病毒SGRNA库，该库的目标是AIM 1中确定的重叠DEG。 CRISPRI和CRSIPRA机器中的10个命中基因将受到基于结构的虚拟屏幕（SBV）的约束 发现可以减轻IPSC-VOC中TKI-VT表型的候选化合物。在AIM 3，3D IPSC- 设计的血管组织（EVT）和小鼠模型将用于验证缓解效率 TKI-VT上的候选化合物在更加物理的环境中。由体内血浆蛋白质组由 人类全血液渗透的3D IPSC-EVT将与患者发现可靠的患者相关 与疾病相关的生物标志物在预测个别癌症患者对TKI-VT的敏感性方面。研究和 在DRS的心态下，K99阶段的职业发展培训计划。 Wu和Ky，以及 专业的跨学科咨询委员会将为Shen博士提供干细胞的高级知识 生物学，血管生物学，心脏肿瘤，CRISPR技术和生物工程。发展的发展 CRISPRI/A和SBVS屏幕平台（K99）和3D IPSC-EVT（R00）将使他能够进行疾病 心脏肿瘤学的建模和药物发现研究特别是血管疾病。新的 在这个K99/R00职业发展奖中获得的技能和经验，加上Shen博士的先前 血管生物学方面的专业知识将促进他向独立职业的过渡，从事基本和 心脏肿瘤学的转化研究特别关注血管方面。