Macrophage-targeted lncRNA-regulating nanoparticles for glioblastoma treatment

巨噬细胞靶向 lncRNA 调节纳米颗粒用于胶质母细胞瘤治疗

基本信息

批准号：
10701432
负责人：
Edjah Nduom
金额：
$ 39.13万
依托单位：
EMORY 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：
2023-09-21 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10701432
关键词：
Acceleration Affect Brain Neoplasms CD14 gene Cancer Patient Cell physiology Cells Clinical Trials Data Development Disease Progression Flow Cytometry Future Genetic Materials Genetic Transcription Genetic study Genetically Engineered Mouse Glioblastoma Goals Immune Immune response Immune system Immunologic Stimulation Immunosuppression Immunotherapeutic agent Immunotherapy Infiltration Interleukin-13 Interleukin-4 Intracranial Neoplasms Knowledge Literature Macrophage Magnetic Resonance Imaging Malignant neoplasm of brain Mediating Modeling Mus Myeloid Cells Operative Surgical Procedures Outcome Patient-Focused Outcomes Patients Peripheral Blood Mononuclear Cell Persons Phenotype Population Radiation Research Resistance Science Small Interfering RNA Sorting Suspensions Therapeutic Tissues Treatment Efficacy Tumor Promotion Tumor-associated macrophages Untranslated RNA Work base bioinformatics tool cancer therapy candidate identification cell type chemotherapy clinical development differential expression immunoregulation improved improved outcome innovation monocyte mouse model nanoparticle nanoparticle delivery neoplastic cell novel novel therapeutics overexpression peripheral blood pre-clinical response standard care theranostics therapeutic target transcriptome sequencing treatment strategy tumor tumor progression tumor-immune system interactions

项目摘要

There is a critical need to identify mechanisms whereby abnormally regulated lncRNAs in macrophages drive progression of GBM and ways to target these lncRNAs preclinically. Our long-term goal is to accelerate the development of immune therapeutics targeting dysregulated lncRNAs to improve outcomes of GBM patients. Our overall objective for this application is to establish a delivery platform targeting novel GBM-specific lncRNAs that promote tumor progression. Our central hypothesis is that overexpression of lncRNAs in TAMs promote GBM tumor progression and immune suppression and are targetable with nanoparticles. The rationale for the proposed research is that evidence of nanoparticle-based targeting of dysregulated GBM-specific lncRNAs will provide new opportunities for the continued development of novel treatment strategies focused on inhibiting TAM-mediated immune suppression. Aim 1: Identify candidate lncRNAs mediating GBM progression. Our approach will be to perform deep bulk RNASeq of CD14+ cell subsets sorted from single cell suspensions of fresh GBM tissue and compare these to the peripheral blood of GBM patients and normal controls to identify differentially expressed lncRNAs, then use bioinformatic tools to choose lncRNAs that are likely to contribute to GBM immune suppression. Our working hypothesis is that TAMs overexpress lncRNAs associated with promotion of immune suppression in GBM beyond those identified from PBMCs alone. Aim 2. Determine the effect(s) of candidate lncRNA depletion on TAM phenotype and function. Our approach will be to deplete candidate lncRNAs in monocytes using siRNA. We will functionally characterize the response to depletion of these lncRNAs and begin mechanistic characterization of novel lncRNAs from Aim 1. Our working hypothesis is that depleting target lncRNAs will change the phenotype and function of these cells, making them resistant to M2-like polarization by IL-4 and IL-13, maintaining a more immune-stimulating state. Aim 3. Develop lncRNA targeted theranostic nanoparticles for lncRNA depletion in TAMs. Our approach will be to further develop MRI trackable nanoparticles for depletion of our preliminary target lncRNAs and any lncRNAs identified in Aim 1 in our genetically engineered murine model of GBM. Our working hypothesis is that our nanoparticles will deplete M2-like TAMs in the murine tumors and improve survival in this model. Our contribution here is expected to be the identification of novel lncRNAs with functional relevance in GBM TAMs and a translational platform for their delivery in murine models of GBM. These contributions will be significant because they are expected to represent an important next step toward future development and clinical trials of novel immune therapeutics for patients with this devastating brain cancer. The proposed research is innovative, in our opinion, because it represents a substantive departure from the status quo by focusing on discovery of immune-related lncRNAs that have been identified from GBM patient myeloid cells and by using nanoparticles to modulate the immune system by targeting these lncRNAs.

迫切需要确定巨噬细胞中异常调节的 lncRNA 驱动的机制 GBM 的进展以及临床前靶向这些 lncRNA 的方法。我们的长期目标是加速开发针对失调的 lncRNA 的免疫疗法，以改善 GBM 患者的预后。我们此应用程序的总体目标是建立一个针对新颖的 GBM 特定的交付平台促进肿瘤进展的lncRNA。我们的中心假设是 TAM 中 lncRNA 的过度表达促进 GBM 肿瘤进展和免疫抑制，并且可以用纳米颗粒靶向。理由对于拟议的研究，证据是基于纳米颗粒的靶向失调的 GBM 特异性 lncRNA将为持续开发新的治疗策略提供新的机会，重点关注抑制 TAM 介导的免疫抑制。目标 1：识别介导 GBM 的候选 lncRNA 进展。我们的方法是对从单个细胞中分选出来的 CD14+ 细胞子集进行深度批量 RNASeq 新鲜 GBM 组织的细胞悬液，并将其与 GBM 患者和正常人的外周血进行比较对照来识别差异表达的lncRNA，然后使用生物信息学工具来选择可能有助于 GBM 免疫抑制。我们的工作假设是 TAM 过度表达 lncRNA 与 GBM 中免疫抑制的促进作用相关，超出了仅从 PBMC 中鉴定出的免疫抑制作用。目标2。确定候选 lncRNA 耗尽对 TAM 表型和功能的影响。我们的方法将使用 siRNA 来消除单核细胞中的候选 lncRNA。我们将从功能上描述响应耗尽这些 lncRNA 并开始对目标 1 中的新型 lncRNA 进行机械表征。工作假设是消耗靶lncRNA会改变这些细胞的表型和功能，使它们能够抵抗 IL-4 和 IL-13 引起的 M2 样极化，从而保持更具免疫刺激的状态。目标 3. 开发 lncRNA 靶向治疗诊断纳米粒子，以消除 TAM 中的 lncRNA。我们的方法将进一步开发 MRI 可追踪纳米粒子，以耗尽我们的初步目标 lncRNA 和任何在我们的 GBM 基因工程鼠模型中，目标 1 中发现了 lncRNA。我们的工作假设是我们的纳米颗粒将耗尽小鼠肿瘤中的 M2 样 TAM，并提高该模型的生存率。我们的这里的贡献预计是鉴定在 GBM TAM 中具有功能相关性的新型 lncRNA 以及用于在 GBM 小鼠模型中传递的转化平台。这些贡献将是巨大的因为它们有望代表未来开发和临床试验的重要下一步针对这种毁灭性脑癌患者的新型免疫疗法。拟议的研究是我们认为，这是创新的，因为它代表着对现状的实质性背离，重点关注发现从 GBM 患者骨髓细胞中鉴定出的免疫相关 lncRNA，并使用纳米颗粒通过靶向这些 lncRNA 来调节免疫系统。