Platelet PI3Kbeta regulation of metastasis

血小板 PI3Kbeta 对转移的调节

• 批准号: 10645152
• 负责人: Ryan C Graff
• 金额: $ 5.27万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10645152
• 关键词: Adhesions; Animals; Antibodies; Antineoplastic Agents; Arteries; Binding; Biological; Biosensor; Blood; Blood Circulation; Blood Platelets; Breast Cancer Cell; Breast Carcinoma; Breast Epithelial Cells; Breast cancer metastasis; Cancer Patient; Cause of Death; Cell Communication; Cells; Collagen; Complex; Data; Distant; Endothelium; Epithelium; Extravasation; Fibrin; Fibrinolytic Agents; Genetic Transcription; Hemorrhage; Hemostatic function; Human; In Vitro; Injections; Intravenous; Invaded; Knock-in; Label; Leukocytes; LoxP-flanked allele; Lung; MCF10A cells; Malignant Epithelial Cell; Malignant Neoplasms; Mammary Neoplasms; Measures; Mediating; Mediator; Megakaryocytes; Mesenchymal; Morbidity - disease rate; Mus; Mutant Strains Mice; NF-kappa B; Neoplasm Circulating Cells; Neoplasm Metastasis; Occlusion injury; PF4 Gene; PIK3CG gene; PTEN gene; Patients; Peptides; Phenotype; Phosphatidylinositols; Phosphotransferases; Platelet Activation; Property; Protein Isoforms; Regulation; Role; Signal Transduction; Site; Solid Neoplasm; Stromal Cells; Testing; Thrombin; Thrombocytopenia; Thrombus; Time; Transcriptional Activation; Tumor Cell Invasion; Tumor Cell Line; Western Blotting; Wild Type Mouse; Work; cancer type; chemokine; cytokine; defined contribution; experimental study; immune clearance; in vivo; inhibitor; matrigel; migration; mortality; mutant; neoplastic cell; platelet function; prevent; promoter; response; stemness; thrombotic; transcriptome sequencing; tumor; wortmannin

Abstract Metastasis is the primary cause of morbidity and mortality among patients with solid tumors. Platelets interact with tumor cells upon their entry into the vasculature and promote the metastatic spread of these cells by several mechanisms. Circulating tumor cells (CTCs) directly bind to and activate platelets resulting in the formation of platelet-fibrin complexes that envelope circulating tumor cells and protect them from immune clearance. Adhesion to tumor cells also induces the release of platelet cytokines and other soluble factors that promote epithelial-mesenchymal transition in the tumor cells. Finally, platelets promote the adhesion of CTCs to the endothelium, assisting in their extravasation at distant metastatic sites. Platelet activation, adhesion, and in vivo thrombus formation require the activity of the Class IA PI 3-kinase PI3Kb. This isoform of PI3K produces the majority of PIP3 in platelets, suggesting that PI3Kb has a unique role in classical platelet activation. However, the role of PI3Kb in platelet-mediated cancer metastasis has yet to be defined. We propose that PI3Kb is required for platelet activation upon interaction with tumor cells, and thereby contributes to platelet-stimulated tumor metastasis. Aim 1 will test the role of platelet PI3Kb in platelet-stimulated tumor cell Matrigel invasion, transendothelial migration, epithelial-mesenchymal transition, cell stemness, NFkB activation, and platelet TGFb secretion. We will also use RNAseq and antibody arrays to more broadly examine the role of PI3Kb in platelet chemokine/cytokine release and in transcriptional responses by tumor cells. We will use both platelets from mice expressing mutant PI3Kb and human platelets pre-treated with the irreversible pan- PI3K inhibitor wortmannin. Aim 2 will directly evaluate the role of platelet PI3Kb in cancer metastasis in wild type and whole-animal or platelet-specific mutant PI3Kb mice. Murine mammary carcinoma cells will be injected intravenously to analyze in vivo tumor cell-platelet complex formation as well as tumor cell interactions with other leukocytes. We will also test the role of PI3Kb in tumor cell-induced thrombocytopenia and experimental metastasis. Isoform selective PI3Kb inhibitors were originally developed to be used as anti-thrombotic agents, as they prevent thrombotic occlusion of injured arteries without increasing bleeding. More recently these inhibitors have also been explored as anti-cancer agents in the treatment of PTEN-deficient cancers. This proposal seeks to understand the potential for a broader application of these agents in cancer patients to inhibit platelet activity and prevent metastasis in a variety of cancer types.

摘要 转移是实体瘤患者发病率和死亡率的主要原因。血小板相互作用 与肿瘤细胞进入脉管系统，并促进这些细胞的转移性扩散， 机制等循环肿瘤细胞（CTC）直接结合并激活血小板，导致血小板形成。 血小板-纤维蛋白复合物包裹循环肿瘤细胞并保护它们免受免疫清除。 与肿瘤细胞的粘附也诱导血小板细胞因子和其他可溶性因子的释放，这些因子促进肿瘤细胞的增殖。 肿瘤细胞中的上皮-间质转化。最后，血小板促进CTC粘附到细胞表面。 内皮，协助其在远处转移部位的外渗。 血小板活化、粘附和体内血栓形成需要IA类PI 3-激酶的活性 PI3Kb。PI 3 K的这种亚型在血小板中产生大部分PIP 3，这表明PI 3 Kb具有独特的作用 在经典的血小板活化中。然而，PI 3 Kb在血小板介导的癌症转移中的作用还有待进一步研究。 定义了我们认为，PI 3 Kb是血小板与肿瘤细胞相互作用后活化所必需的，因此， 有助于血小板刺激的肿瘤转移。目的1检测血小板PI 3 Kb在血小板刺激的 肿瘤细胞Matrigel侵袭，跨内皮迁移，上皮-间质转化，细胞干细胞，NF κ B 活化和血小板TGF β分泌。我们还将使用RNAseq和抗体阵列更广泛地检查 PI 3 Kb在血小板趋化因子/细胞因子释放和肿瘤细胞转录应答中的作用。我们将 使用来自表达突变型PI 3 Kb的小鼠的血小板和用不可逆泛- PI 3 K抑制剂渥曼青霉素。目的2直接评价血小板PI 3 Kb在野生型肺癌转移中的作用 和全动物或血小板特异性突变体PI 3 Kb小鼠。将注射小鼠乳腺癌细胞 静脉内注射以分析体内肿瘤细胞-血小板复合物形成以及肿瘤细胞与其它细胞的相互作用。 白细胞我们还将测试PI 3 Kb在肿瘤细胞诱导的血小板减少症和实验性血小板减少症中的作用。 转移 同种型选择性PI 3 Kb抑制剂最初被开发用作抗血栓形成剂，因为它们 防止损伤动脉的血栓性闭塞而不增加出血。最近，这些抑制剂 也被探索作为治疗PTEN缺陷型癌症的抗癌剂。这项建议旨在 了解这些药物在癌症患者中更广泛应用以抑制血小板活性的潜力 并防止多种癌症类型的转移。