Targeting microtubule stabilization to reduce breast tumor metastasis

靶向微管稳定以减少乳腺肿瘤转移

• 批准号: 10212975
• 负责人: STUART S MARTIN
• 金额: $ 36.69万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-06 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212975
• 关键词: Acetylation; Affect; Animal Model; Antineoplastic Agents; Behavior; Biomedical Engineering; Blood Circulation; Blood capillaries; Breast Cancer Cell; Breast Cancer Treatment; CRISPR/Cas technology; Cells; Cessation of life; Characteristics; Chemicals; Clinical Treatment; Communities; Confocal Microscopy; Curcumin; Data; Disease; Distant; Drug Targeting; Electron Microscopy; Extracellular Matrix; FDA approved; Foundations; Genetic; Growth; HDAC6 gene; Histone Deacetylase; Histone Deacetylase Inhibitor; Hour; Human; Image; In Vitro; Invaded; Ixabepilone; Lung; Lymphatic; Mammary Neoplasms; Mechanics; Metastatic breast cancer; Methods; Microfluidics; Microtubule Stabilization; Microtubules; Modification; Molecular; Mus; Neoadjuvant Therapy; Neoplasm Circulating Cells; Neoplasm Metastasis; Operative Surgical Procedures; Organ; Outcome; Paclitaxel; Patients; Pharmaceutical Preparations; Phenotype; Post-Translational Protein Processing; Prognosis; Recurrence; Risk; Role; Structure; Surface; System; Testing; Therapeutic; Tissues; Translating; Treatment Efficacy; Tubulin; Xenograft procedure; analog; anticancer research; base; cancer therapy; cell growth; cell motility; chemotherapy; drug development; drug discovery; drug testing; efficacy testing; epithelial to mesenchymal transition; improved; in vivo; innovation; interest; molecular marker; mouse model; neoplastic cell; novel; novel strategies; novel therapeutics; parthenolide; patient derived xenograft model; precision medicine; pressure; response; side effect; standard of care; stem cells; therapeutic evaluation; therapeutic target; treatment strategy; tumor; whole animal imaging

Targeting microtubule stabilization to reduce breast tumor metastasis. Breast tumor cells metastasize to distant organs through non-adherent microenvironments, such as the bloodstream or lymphatics. However, very little is known about the dynamic behavior and drug responses of non-adherent tumor cells, due to the challenges of imaging non-adherent cells without blurring from cell drift. The PI's lab discovered unique microtentacles (McTNs) on the surface of non-adherent tumor cells that promote the aggregation and retention of circulating tumor cells (CTCs) in the lung capillaries of living mice. This revised study will test the hypothesis that therapeutic targeting of 2 microtubule stabilization mechanisms (detyrosination, acetylation) can provide a novel strategy to suppress tumor metastasis and move beyond the current non-selective MT-targeting drugs that are aimed at tumor cell growth. Predictions of this hypothesis will be tested in the following specific aims with confocal and electron microscopy, whole-animal imaging of CTC metastasis and by testing drugs on patient tumor cells and CTCs. Specific Aim 1: Inhibit tubulin detyrosination to reduce McTNs, stem cell characteristics and metastasis. A) Alter tubulin detyrosination by targeting TTL or TCP and gauge effects on metastatic potential/EMT. B) Test efficacy of Curcumin and Parthenolide analog on EMT, stem cell characteristics and metastasis. C) Define whether microtubule-stabilizing therapies (Paclitaxel, Ixabepilone) promote metastatic potential. Specific Aim 2: Determine the effects of altering tubulin acetylation on metastatic phenotypes. A) Genetically alter tubulin acetylation (ATAT1, K40R, HDAC6) and gauge effects on metastasis in mice. B) Analyze tumor cell mechanics and McTN structure in cells with altered tubulin acetylation. C) Examine whether elevation of tubulin acetylation by HDAC inhibition increases metastatic potential. Specific Aim 3: Target microtubule stabilization mechanisms in live CTCs from mice and human patients. A) Test 2 prioritized drugs on xenografts, PDX and live patient CTCs to reduce McTNs and CTC clusters. B) Compare CTC metastasis with MT-stabilizing and MT-disrupting agents in isolation and combination. An innovative microfluidic cell tethering system will be used to rapidly determine drug responses in live patient CTCs in less than one hour. Inclusion of numerous FDA-approved therapies will increase the potential to rapidly translate the outcomes of this project to impact the clinical treatment of metastatic breast cancer.

靶向微管稳定以减少乳腺肿瘤转移。 乳腺肿瘤细胞通过非粘附性微环境转移到远处器官，如 血液或血液循环。然而，很少有人知道的动态行为和药物反应， 非粘附性肿瘤细胞，这是由于在不因细胞漂移而模糊的情况下对非粘附性细胞进行成像的挑战。 PI的实验室在非粘附肿瘤细胞表面发现了独特的微触角（McTN）， 促进循环肿瘤细胞（CTC）在活小鼠肺毛细血管中的聚集和滞留。 这项修订后的研究将检验这一假设，即治疗靶向2微管稳定 机制（去酪氨酸、乙酰化）可以提供抑制肿瘤转移的新策略 并超越目前针对肿瘤细胞生长的非选择性MT靶向药物。 这一假设的预测将在以下具体目标与共焦和电子测试 通过显微镜、CTC转移的整体动物成像以及通过在患者肿瘤细胞和CTC上测试药物来进行。 具体目标1：抑制微管蛋白脱酪氨酸，以减少McTN、干细胞特性和转移。 A）通过靶向TTL或TCP改变微管蛋白脱酪氨酸，并测量对转移潜能/EMT的影响。 B）测试姜黄素和银胶菊类似物对EMT、干细胞特征和转移的功效。 C）确定微管稳定疗法（Paclitazone，Ixabepilone）是否促进转移潜力。 具体目标2：确定改变微管蛋白乙酰化对转移表型的影响。 A）在小鼠中遗传改变微管蛋白乙酰化（ATAT 1、K40 R、HDAC 6）并测量对转移的影响。 B）分析具有改变的微管蛋白乙酰化的细胞中的肿瘤细胞力学和McTN结构。 C）检查通过HDAC抑制提高微管蛋白乙酰化是否增加转移潜能。 具体目标3：小鼠和人类患者活CTC中的靶向微管稳定机制。 A）测试2优先考虑异种移植物、PDX和活患者CTC上的药物以减少McTN和CTC簇。 B）将CTC转移与MT稳定剂和MT破坏剂单独和组合进行比较。 创新的微流控细胞系留系统将用于快速确定活患者的药物反应 一小时内完成CTC。纳入众多FDA批准的疗法将增加 迅速将该项目的成果转化为影响转移性乳腺癌的临床治疗。