Imaging Modulation of Immune Phenotype

免疫表型的成像调节

• 批准号: 10548151
• 负责人: Katherine W Ferrara
• 金额: $ 64.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10548151
• 关键词: Abbreviations; Abraxane; Acids; Address; Adverse event; Agonist; Animal Model; Animals; Biological Assay; Biopsy; Cancer Etiology; Cells; Characteristics; Chemistry; Clinical Treatment; Clinical Trials; Combined Modality Therapy; Complex; Data; Dendritic Cells; Detection; Development; Diagnosis; Dose; Drug Kinetics; Environment; Esterification; Extravasation; Flow Cytometry; Formulation; Goals; Head and Neck Cancer; Image; Immune; Immune response; ImmunoPET; Immunophenotyping; Immunotherapy; Implant; In Vitro; Interferons; Kidney; Lectin; Leukocytes; Lymphoma; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Mannose; Metals; Methods; Modeling; Modernization; Monitor; Monoclonal Antibodies; Mus; Myeloid-derived suppressor cells; Nature; OX40; PADRE 45; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Peptides; Phenotype; Polymers; Positron-Emission Tomography; Prognosis; Property; Protocols documentation; Reaction; Reporting; Resistance; Rodent; Safety; Sampling; Site; T cell response; T-Cell Activation; T-Lymphocyte; TLR7 gene; TNFRSF5 gene; Techniques; Testing; Therapeutic; Therapeutic Index; Time; Toll-like receptors; Toxic effect; Translations; Unresectable; Update; Work; adaptive immune response; anti-CTLA4; anti-PD-1; biomaterial compatibility; checkpoint therapy; chemotherapy; efficacy evaluation; gastrointestinal; gemcitabine; immunoregulation; improved; in vivo; lipophilicity; malignant breast neoplasm; mortality; mouse model; nanoformulation; nanoparticle; nanoparticle delivery; nanotherapeutic; nanotherapy; non-invasive monitor; pancreatic cancer cells; pancreatic cancer model; pancreatic cancer patients; pancreatic neoplasm; particle; preclinical study; predicting response; receptor; resiquimod; response; scaffold; small molecule; specific biomarkers; transcriptome sequencing; translation to humans; treatment optimization; tumor; tumor microenvironment; uptake

Imaging and modulation of immunophenotype Most importantly, we have created TLR delivery nanotherapeutics and find that TLR NPs combined with aCD40, aPD-1 and aCTL4 (abbreviated as CP4 as in emerging pancreatic cancer studies) efficiently regressed implanted multisite invasive murine pancreatic tumors. We have developed multiple strategies and are particularly focused on 18 nm biodegradable, multi-functional particles that combine immune-modulating peptides, targeting peptides and toll-like receptor (TLR) agonists. We specifically included the immune modulating peptide PADRE (T helper modulation) and mannose (to enhance macrophage uptake) in addition to the TLR7/8 agonist (resiquimod) in preliminary work. To maximize payload, we built upon biocompatible unimicellar nanoparticles via the combination of highly efficient esterification and metal-free click reactions and find that the particle metabolites clear through the kidneys. In our preliminary studies, TLR7/8-nanoparticle treatment combined with CP4 enhanced response in a highly metastatic, multi-site implanted pancreatic cancer model (Kras+/LSL-G12D; Trp53+/LSL-R172H; Pdx1-Cre model: abbreviated as KPC). New preliminary data indicate that TLR7/8 agonists and aCD40 each have direct efficacy against pancreatic tumor cells. RNAseq results demonstrate that TLR7/8 agonists and CD40 enhance complementary pathways (C-lectin for CD40 (among others) and TLR/interferon for TLR agonists). We find that the combination enhances anti-tumor leukocytes, regresses KPC tumors and for responders, 100% do not grow tumor on re-challenge. By monitoring OX40 expression (a marker of T cell activation), we demonstrated that unlike other immune modulating approaches involving aCD40, T cells were activated. We have simultaneously developed the ability to monitor OX40 expression using positron emission tomography in a noninvasive fashion. Our primary goal in the proposed work is to move the nanotherapy strategy forward to human translation. As a result, we will evaluate efficacy in models of pancreatic cancer in rodents and safety in a larger animal model. Further, we will evaluate samples from patients undergoing biopsy for pancreatic cancer to better characterize the immune environment. We have 2 major goals: 1) the development of an effective strategy for systemically-administered T cell modulation and 2) combining this with positron emission tomographic imaging and RNA sequencing to optimize multi-component protocols. Within Aim 1, we will determine the optimal carrier properties to maximize T cell modulation by 1a) modulating nanoparticle characteristics and evaluating resulting efficacy, 1b) using positron emission tomography (PET) imaging to quantify accumulation of the systemically-injected NP agonists, and 1c) assessing toxicity through dose escalation and a large animal study, leading to IND filing. Within Aim 2, develop an imaging and in vitro assessment strategy for T cell activation by utilizing 2a) OX40 PET imaging and 2b) flow cytometry and RNAsequencing.

免疫表型的成像和调节 最重要的是，我们已经创造了TLR递送纳米治疗剂，并发现TLR NP与aCD 40结合， aPD-1和aCTL 4（在新兴胰腺癌研究中缩写为CP 4）有效地消退了植入 多部位侵袭性鼠胰腺肿瘤。我们制定了多项战略， 在18 nm生物可降解的多功能颗粒上，该颗粒结合了联合收割机免疫调节肽、靶向肽 和Toll样受体（TLR）激动剂。我们特别包括免疫调节肽PADRE（T辅助细胞 调节）和甘露糖（以增强巨噬细胞摄取），以及TLR 7/8激动剂（瑞喹莫特）。 前期工作为了最大限度地提高有效载荷，我们建立了生物相容性单胶束纳米粒子通过 高效酯化和无金属点击反应的组合，并发现颗粒代谢物 通过肾脏排出在我们的初步研究中，TLR 7/8-纳米颗粒治疗结合CP 4 在高转移性、多部位植入的胰腺癌模型中增强的反应（Kras+/LSL-G12 D; Trp 53 +/LSL-R172 H; Pdx 1-Cre模型：缩写为KPC）。新的初步数据表明，TLR 7/8激动剂和 aCD 40各自对胰腺肿瘤细胞具有直接功效。RNAseq结果表明，TLR 7/8 激动剂和CD 40增强互补途径（C-凝集素的CD 40（除其他外）和TLR/干扰素 对于TLR激动剂）。我们发现该组合增强了抗肿瘤白细胞，使KPC肿瘤消退， 应答者，100%在再激发时不生长肿瘤。通过监测OX 40表达（T细胞标志物 活化），我们证明了与其他涉及aCD 40的免疫调节方法不同，T细胞是 激活我们同时开发了使用正电子发射来监测OX 40表达的能力。 无创性断层扫描我们在拟议工作中的主要目标是将纳米治疗策略 转向人工翻译。因此，我们将在啮齿动物胰腺癌模型中评估疗效 和安全性。此外，我们将评估接受活检的患者的样本， 胰腺癌，以更好地表征免疫环境。我们有两个主要目标：1）发展 一种有效的策略，用于系统管理的T细胞调节和2）结合这与正电子 发射断层成像和RNA测序，以优化多组分方案。在目标1中，我们 将确定最佳载体性质以通过以下方式最大化T细胞调节：1a）调节纳米颗粒 1b）使用正电子发射断层扫描（PET）成像， 量化全身注射的NP激动剂的累积，和1c）通过剂量评估毒性 升级和大型动物研究，导致IND备案。在目标2中，开发成像和体外 通过利用2a）0X 40 PET成像和2b）流式细胞术评估T细胞活化的策略，以及 RNA测序。