Targeting Glioblastoma with a Nuclear-penetrating Anti-DNA Autoantibody

使用核穿透性抗 DNA 自身抗体靶向胶质母细胞瘤

• 批准号: 10362737
• 负责人: James E. Hansen
• 金额: $ 36.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10362737
• 关键词: Adenosine; Agonist; Antinuclear Antibodies; Area; Autoantibodies; Binding; Biometry; Biophysics; Blood - brain barrier anatomy; Brain; Cardiotoxicity; Cell Nucleus; Cells; Clinical; Clinical Trials; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; Development; Dose; Double Strand Break Repair; Drug Delivery Systems; Drug Kinetics; Engineering; Environment; Evaluation; Foundations; Genetic; Genetic Heterogeneity; Glioblastoma; Growth; Human; Immunocompetent; Immunodeficient Mouse; Intercellular Junctions; Ligands; Lupus; Malignant Neoplasms; Methods; Modeling; Mus; Mutation; Normal Cell; Nuclear; Nucleoside Transporter; Nucleosides; Operative Surgical Procedures; PTEN gene; Pathway interactions; Penetration; Pharmaceutical Preparations; Pharmacology; Pilot Projects; Purinergic P1 Receptors; Radiation; Radiation therapy; Radiation-Sensitizing Agents; Relapse; Research Personnel; Risk; Role; Site; Surface; Systemic Lupus Erythematosus; Technology; Testing; Toxic effect; Translating; Work; aggressive therapy; blood-brain barrier penetration; blood-brain barrier permeabilization; brain endothelial cell; cancer cell; chemotherapy; drug development; efficacy evaluation; experience; experimental study; extracellular; improved; in vivo; innovation; mortality; mouse model; nanocarrier; neoplastic cell; neuro-oncology; novel; novel strategies; novel therapeutics; patient derived xenograft model; restoration; risk minimization; standard of care; stem-like cell; systemic autoimmune disease; tumor; tumor growth; uptake

Glioblastoma multiforme (GBM), the most common primary malignancy of the brain, has high rates of relapse and mortality despite aggressive treatment with surgery, radiation, and chemotherapy. Improved methods to target and treat GBM are needed, but the blood-brain barrier (BBB) and genetic heterogeneity associated with GBM are obstacles to drug development. The autoimmune disease systemic lupus erythematosus offers an unexpected new approach. We discovered that the lupus anti-DNA autoantibody 3E10 penetrates live cell nuclei and inhibits DNA repair in a manner that does not kill normal cells but is toxic to cancer cells with genetic defects in repair of DNA double- strand breaks (DSBs), including PTEN-deficient cancer cells and tumors. 3E10 penetrates cells via a mechanism that requires both extracellular DNA and the ENT2 nucleoside transporter to be present, which facilitates preferential penetration of 3E10 into tumors due to their expression of ENT2 and increased DNA in their environment. ~40% of primary GBM is PTEN-deficient, and we hypothesize that 3E10 will target GBM, have single agent effects against PTEN-deficient GBM, and can serve as a sensitizing agent for radiation therapy and as a drug delivery ligand for GBM regardless of PTEN status. We have re-engineered a 3E10 fragment, hereafter referred to as Deoxymab-1 (DX1), to maximize effect on cancer cells and minimize risks. In preliminary studies DX1 crosses the BBB to localize into and suppress growth of PTEN-deficient GBM in an orthotopic patient-derived xenograft (PDX) mouse model, and delivers conjugated nanocarriers to orthotopic GBM tumors. We now propose studies to help translate DX1 into a novel therapy for GBM. In Aim 1 we pursue studies to elucidate and enhance the mechanism by which DX1 crosses the BBB in GBM. In Aim 2 the effects of DX1, alone or in combination with radiation therapy, on viability and DNA damage accumulation in GBM and normal cells will be determined. DX1 +/- radiation therapy will then be tested in PDX and syngeneic mouse models of GBM to evaluate efficacy and toxicity. In Aim 3 methods to deliver drug-loaded nanocarriers by surface conjugation with DX1 are developed and tested in orthotopic GBM models. We believe use of a modified nuclear-penetrating lupus anti-DNA autoantibody against GBM is an innovative and compelling new strategy that has potential for significant clinical impact, and the proposed studies will establish a foundation for advancing the DX1 technology to clinical trials.

胶质母细胞瘤多形（GBM）是大脑中最常见的主要原发性恶性肿瘤，具有较高的复发率 尽管对手术，放射线和化学疗法进行了积极治疗，但死亡率和死亡率。改进的方法 需要目标和治疗GBM，但是血脑屏障（BBB）和遗传异质性与 GBM是药物开发的障碍。自身免疫性疾病全身性红斑狼疮提供 意外的新方法。 我们发现狼疮抗DNA自身抗体3E10穿透活细胞核并抑制DNA修复 一种不杀死正常细胞但对具有遗传缺陷的癌细胞有毒的方式 链断裂（DSB），包括缺陷型PTEN癌细胞和肿瘤。 3E10通过机制穿透细胞 这需要存在细胞外DNA和ENT2核苷转运蛋白，这促进了 3E10的优先渗透到肿瘤中，因为它们的ENT2表达并增加了DNA 环境。 〜40％的主要GBM缺乏PTEN，我们假设3E10目标GBM具有 对PTEN缺乏GBM的单一药物影响，可以作为放射治疗和 无论PTEN身份如何，GBM的药物输送配体。 我们已经重新设计了一个3e10片段，以下称为脱氧符1（dx1），以最大程度地提高对对的影响。 癌细胞并最大程度地降低风险。在初步研究中 原位患者衍生的异种移植物（PDX）小鼠模型中缺乏PTEN缺乏的GBM，并提供共轭 原位GBM肿瘤的纳米载体。现在，我们提出研究，以帮助将DX1转化为一种新的疗法 GBM。在AIM 1中，我们进行研究以阐明和增强DX1越过BBB的机制 GBM。在AIM 2中，DX1的影响，单独或与放射治疗，对活力和DNA损伤结合 将确定在GBM和正常细胞中的积累。然后将在PDX中测试DX1 +/-辐射疗法 和GBM的合成小鼠模型，以评估功效和毒性。在AIM 3方法中输送毒品的方法 通过与DX1进行表面结合的纳米载体在原位GBM模型中开发并测试。 我们认为，使用改良的核穿透狼疮抗DNA自动抗体针对GBM是一种创新的 并引人注目的新策略有可能产生重大临床影响，拟议的研究将 建立将DX1技术推进临床试验的基础。